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NATURAL HISTORY

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THE BULLETIN

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allege of Agriculture.

TOKYO IMPERIAL UNIVERSITY,

JAPAN.
Vol. II.

1894-1897.

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CONTENTS OF VOLUME II.

Pace.

The Energy of the Living Protoplasm. By Prof. Oscar
Lorw Gm fi : si, (6 ox pee de. ky 435% 150:

On the Poisonous Action of eee By Prof. O. Lozw
and M. Tsuxamoto, Mogakusht. .. .. .. .. . pee-Claees ye

On the Vegetable Cheese, Na/oo. By K. Yasr, Mogakusht. .. 68.

On the Poisonous Action of the Hydroxyl-derivatives of
Benzol upon Yeast and Bacteria. ByK. Yas, Mgakusht. 73.

On the Quantity of Wood-Gum (Xy/an) Contained in
Different Kinds of Wood. By J. Oxumura, Mogakusht. .. 76.

On the Reserve Proteinin Plants. By G. Darkunara, Mogakusht. 79.
On the Occurence of Mucinin Plants. By J. Isuu, Nogakusht. 97.
Iiannane as a Reserve Material in the Seeds of Diospyros

aka is. (ost ISHIL, IWeakusht, 0 eS EOL.
Mannane as an Article il Human Food. By C. Tsvuyl,
Nogakushi. a) = 103°

On the Scale Insect of raibaery Trees | (Diaspis Sabet
formis,) (Plates I-II.) he Prof. C. Sasaki, Regwku-

RGRUSHD ss Plakis Ween Boe 1O7:
On the Ss conietia a of the Silk-Worm. (Plates III-iV.)
ame OVA A GSMS eM Lie ie) ice Pras ey aay foe TRS:

On the Reserve Protein in Plants. II. By G. Darxun,ra,
OGRA: SA Me ie oy tO es Fa 5. TOO:
On the Consumption of Asparagine in the Nutrition of
Plants, By Y. Kinosaira, Wogakusht. .. .. .. .. «.. 196.
On the Assimilation of Nitrogen from Nitrates and Am-
monium Salts by Phaenogams. By Y. Kinosuita, M-
CORUS/iim hr a 2 ZOO:
On the Presence ae eee, in the Root of Nelu::bo
nucifera. By. Y. Krinosuita, Nigakushi. LS OT! BMT E2035
On the Occurence of Two Kinds of Mannane in the Root
of Conophallus konyaku. By Y. Kivosuita, Nogakushi. 205.
Wote on the Chemical Composition of some Mucilages.
Byam MoswiMurA, WVogakushi, © <0 <. «+ «0 se 04 0% 207.

eee

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The Preparation and Chemical Composition of TZo/u.
By M. Inouye, Mogahkushii .. .. Poy (we are OOS
Note on Nukamiso. By M. Inouye, Nogabushe Wee
Preliminary Note on the Sake Yeast. By K. Yanr, Nogakusht. 219.

Note on the Behaviour of Hyppurie Acid in Soils. By K.
WosHimura, JVogakushz, .. <2 (acura es eer

Does Hydrogen Peroxide occur in Plants? By J. Cxuo os, B25.

Die Japanischen Laubholzer im Winterzustande. Bes-
timmungstabellen. Von H. Suirasawa, Ringakushi. sv 220)

Untersuchungen uber das Klemmen der Technisch Wich-
tigsten Japanischen Holzarten. Von F. Kor, Ringaku-
7) Sc oS OOP CE cee 2S

Ertragstafel und Zuwachsgesetz fur Sugi (Cryplomeria Fapo-
nica) zum Gebrauch fur die Japanischen Forstmanner.
Von Serroku Honpa, Ringakusht et Dr. Oec. publ... .. .. 335.

Uber den Hinfluss Wechselnder Mengen von Kalk und
Magnesia auf die Entwicklung der Nadelbaume. Von
Prof. Dr. O. Lozw und Prof Dr. S. Honpa. = oa nS

Uber die Entstehung der Verkrummungen an Fotsuya-
maruta (Sugi-Stangenholz). Von Prof. Dr. S. Honpa. .. 387.

Besitzen die Kiefernadeln ein Mehrjahriges Wachstum ?
Von Prof. Dr. 5: HONDA 20 1.) S|

Lability and Energy in Relation to Protoplasm. By Prof.
Dr. O. Loew. ee CCM MEEIESES 65 a BOS

On the Formation of Mannan in Amorphophalus Kowjak.
By M. Tsuxamoro, ogakusht) me, .. .. .@aen) =u

On the Formation of Asparagine in Plants under Different
Conditions. By U. Suzuxt, Mogakusht. .. .. ..... «. 409.

Can Old Leaves of Plants produce — by Starva-
tion? By T. Mivacui, Mogakushr ..° .. . - 459.

On the Relative Value of Asparagine as a Nutrient for
Phaenogams. By T. Nakamura, Mogakusht. «oR 465.

On the Relative Value of Asparagine as a Nutrient for
Fungi. By T. Nakamura, Mogakusht, .. .. . «.. «. 468.

On the Quantities of Nitrates Stored up in Plants under
Different Conditions. By T. Isnizuxa, Migakusht. .. .. 471.

—_ 3 —
On the Significance of the Nitrates Contained in Plants

for Animals and Men. By T. Isuizuxa, Nogakusht. .. .. 475.
On the Physiological Behaviour of Maleic and Fumaric
Acids. By T. Isuizuxa, Mogakusht. .. .. - 484.
On the Physiological Action of Amiosulphonie Acid. By
N. Magno, Mogakusht. .. .. . Og ob ee Neco ae ty
Investigations on the maiperes Tree. By N. Magno. JVWo-
I ORUS hte mele : ert OA.
Notes on the Metabolism in the Cherry Tree wy S.
Aoyama, JVogakushi. Aa ae - 499.

Physiological Observations on Lecithin. By T. Hanat,
UNO SCRUSHE = iy eel anes Wie Seem) Pie is elas. ~ ie sg. 5O35

On a Compound of Albumen with Phenol. By M. Surmapa,
Nogakusht. Gi | aS OE Mgpae SOL COC ECCON, sith OCMC Same OH

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The Energy of the Living Protoplasm.

BY Dr. Oscar Loew.

Professor of Agricultural Chemistry.

CHAPTER I.

FORMER VIEWS ON THE CAUSE OF
THE VITAL PHENOMENA.

The change from life to death is so striking, that it has been
from the oldest times up to the present day considered as a
mystery, as an inconceivable supernatural process. Not less
striking than the death of a larger animal or plant is the death-
moment of the lowest forms of animal or vegetable life, watched
under the microscope. The motions of infusoria cease, the
diatoms stop, the spirogyracells show contraction of their green
spirals, their cytoplasm loses its shape, their nucleus leaves its
normal position. All energy, mechanical or chemical, is gone !—

Living organisms produce /eat, the nervous system electricity,
certain fishes even powerful electrical phenomena, a number of
animals and fungi emanate Jight and all the animals as well as
some forms of lower vegetable organisms are capable of locomo-
tion. And what a great amount of chemical energy is actively
engaged in producing organic matter in plants, on the one hand,
and in decomposing and oxydising it in animals on the other!
All these phenomena stop at the moment of death. But what
is the cause of the disappearance of the forms of energy? It is
of no avail to answer: ‘‘ Respiration produces heat which can
be converted into other forms of energy;”’ here the fundamental
question is: What has produced the respiration process ?. Why does
this process stop at once at the moment of death ?—

Before we enter upon the discussion and scientific treatment
of this question let us glance at the opinions of ancient philoso-
phers and modern physiologists in regard to the cause of the
vital functions.

2 FORMER VIEWS ON THE CAUSE

Anaxagoras defined an animal as an automatic machine, but
he left undecided what cause moved the machinery. Socrates
ridiculed this idea even on his death-bed.) According to Ari-
stotle the animal motions and animal heat are intimately con-
nected; the heat is produced by the food. The heart is the
centre of motion and sensation, has a life of its own, and is the
hottest part of the body.” Plato considered the red color of the
blood as an effect of the life-fire, and the blood itself as the
bearer of the vital force, as the seat of the soul.?) The Pytha-
goreans defined animal life as the result of the entrance of
the ‘‘life-spirit’”” into the body’) by the respiration-process,
and declared the brain to be the seat of sensation.—As
the scientific development ceased in Europe for a very long
time, we do not find any discussions upon this subject up
to Descartes, who in the year 1637 expressed his conviction
that all the forces of nature consist in certain motions of the
molecules; the animals were in his opinion caloric automatic
machines, in which the motions of the blood and of the organs
are the effects of chemical heat-producing processes. The finest
parts of the blood are a kind of nervous ether that ascends from
the heart to the brain.*? Baco and Descartes were the first, who
considered heat as a mode of motion, and Descartes extended his
views also to light, electricity, and magnetism.

With the observation of Galvani in the year 1780 of the
convulsions of a frog’s leg brought in contract with two metals,
the view of Galvani found numerous followers, and men like
Humboldt were among the admirers of the new vital theory.
However Volta’s experiments an contact-electricity soon eclipsed
the results of Galvani, and when the latter succeeded in the
year 1793 in adducing irrefutable proof of the presence of an
electrical current in the animal: electrical contraction without
metals, it did not find grace with the scientific public; the
doubts and distrusts created by Volta were mightier than the

1) Plato, Phaedon 97, 98.

2) Aristotle (Editio Bekkeri) de part. anim. II, 1; III. IV, 4, 7; V. 2.—Hist.
anim. I. XVII.—De respiratione VI.

3) Plato, Tim. 493 etc.

4) Democrit, in: Aristotle, de respiratione IV.

5) René Descartes works, edited by H. Kirchmann, II, 27.

OF THE VITAL PHENOMENA. 3

language of the new experiments; Galvani did not receive any
satisfaction, he died without having found any recognition.
And when Du Bois Reymond proved in the year 1843 the cor-
rectness of Galvani’s views of the presence of electrical currents
in animals, nevertheless no voice was raised in favour of the
theory that electricity is the ‘‘pvimum movens”’ of all the vital
phenomena. And indeed the electrical phenomena observed are
just like the heat secondary actions, but not the first cause
of life.

Since the middle of last century another view has gained
much foothold—, a view that was even defended by Liebig—, the
theory that organisms are ruled by quite a specific force, different
from any other, inscrutable to men, and supernatural: this force
was called vital force. Fustus Liebig said: ‘‘the cause of vital
force is not chemical force, not electricity, not magnetism; it is
a force that possesses the most general qualities of all causes
of motion, of variation of form and qualities of matter, and a
specific force,’ because it produces effects like no other force.”
“The laws of life and every thing disturbing, promoting, or
varying them can doubtless be investigated, but without ever
knowing what life really is.” Even in the third edition of this
work’), published 1846, we find upon the first page: ‘‘in the
animal egg, in the plant-seed is recognizable a remarkable
activity, a cause of increase in substance, a compensation of loss,
aguioceein the state of mesti.4.c:cn0 this force we call vital force.”
From page 225 the following characteristic passage may be
quoted : “ another fundamental error entertained by physiologists
is, that physical or chemical forces alone or in combination with
anatomy could suffice to explain the vital phenomena.”

This reactionary movement of Liebig was principally due to
the opposition to the great and fervent hopes created by the
first synthesis of an organic compound, namely that of urea by
Woehley in the year 1828. Previous to this year it was often
asserted that organic substances could only be formed by “ vital

1) Die organ. Chem. in ihrer Anwendung auf Physiol. und Pathol., Braun-
schweig 1842 p. 7 and p. 237. These few quotations will clearly show how erroneous
the opinion is, that Liebig was the first, who combated the hypothesis of the
supernatural vital force, an opinion, which we find in some german textbooks of
physiological chemistry. See also Chem. Briefe of 1858. Chapt. 23.

4 FORMER VIEWS ON THE CAUSE

force,” a view that was adopted by Berzelius in 1827 in Vol. III
p. 1. of his textbook of chemistry. But even in the year 1847
this prominent chemist declared that life is something mysterious
and inscrutable; “once connected with matter it produces de-
velopment and growth, but how this proceeds is an insoluble
mystery"). Similar declarations were made by Tvevivanus, who
believed that the vital principle passes off into the air with
the death of an organism”). ‘The view that ‘life was not in any
way related to the physical forces and had nothing in common
with any material forces, powers, or properties,” was even defend-
ed quite recently by some learned professors of the Victoria
Institute, London>). ‘‘ Different views however were entertained
nearly a hundred years ago by the German physiologist Rel,” :
“the socalled vital force has fooled us long enough, and has led
us into sterile deserts. Matter itself and not a specific new force
is the cause of vital phenomena.”’ Such was essentially also the
conviction of Oken and of De Candolle, while Mulder>) declared:
“the vital force is a sfecific force connected with the 4 elements:
carbon, hydrogen, nitrogen and oxygen.” —Evlenmeyer, well known
for his numerous investigations in the domain of theoretical or-
ganic chemistry declared®: “all our knowledge of chemistry
and physics has been set in motion to solve the mystery of life,
and we have made a great step forward; but if a physiologist
like C. Ludwig identifies the progress of physiology with the pro-
gress of chemistry, then the chemists must feel anew instigated
to devote themselves to physiological problems.” ‘‘ The vitalistic
school had more power in former times, chemistry and physics
having been too imperfectly developed.’’ To similar opinions
inclined the physiologist Lehmann (1853): ‘‘if many vital phe-
nomena are inexplicable up to the present day, we still do not
feel the necessity of nominating such a governor as vital force.”) ”’
Quite differently expressed himself another physiologist, Gorup-
Besanez (1874): ‘‘ All the physical and chemical laws known to

1) Lehrbuch der Chemie, 5th Edition, Vol. 4, p. 5.

2) Physiologie der Gewachse 1835. Vol. I, p. 12.

3) Compare ‘‘ Science” of March 1893.

4) Reil’s Archiv III, 424 (1798).

5) Versuch einer allg. physiol. Chem. p. 92 (1843).

6) Zeitschrift f. Chem. 1859.

7) Lehrbuch d. physiol. Chem. II, Ed. Vol. III, p. 154.

OF THE VITAL PHENOMENA. 5

us at the present day are not sufficient to explain the formation
of a plantcell, the process of generation or the conduction of the
sense-impressions to the brain.”) ”

Among those who most emphatically denied the necessity for
the hypothesis of a special supernatural force were: Schleiden,
Matteucct, Moleschott, Huxley, Haeckel, R. Mayer, F. Hiippe,
Heidenhain and Halliburton. Schleiden® uttered the following
philippica: “Only ignorance and indolence of spirit are the
defenders of the vital force at the present state of development
of the natural sciences, of a force that can accomplish every-
thing and explain everything, and of which nobody can tell, how
it acts nor what laws it obeys. The savage, who takes a
locomotive for a wild animal, is not more ignorant, than the
natural philosopher who talkes about vital force in organisms.”’

Matteucci : “‘ to explain everything by vital force, and yet not
to know the laws of this supposed force, explains nothing; it is
even worse than nothing, it prevents the mind from searching
after the truth.?) ”

Moleschott : ‘‘ Life is not the result of a specific force, it isa
certain state of matter caused by peculiar motions produced by
heat and light, water and air, electricity and mechanical con-
vulsions.*) ””

Th. Huxley): ‘To speak in what is not altogether a
metaphor, the atoms, which enter the body are for the most part
piled up in large heaps and tumble down into small heaps
before they leave it. The force, which they set free in thus
tumbling down, is the active power of the organism.” Huxley
forgets, however, to mention the cause of the “‘ tumbling down.”
Haeckel takes the question still easier, he finds the formation of
a cell as simple as that of a crystal, and assumes the existence
of sensation and motion in every atom. This “soul” is
modified and perfected by the development of the organisms.®

Very naturally Robert Mayer did not believe in a mysterious

1) Lehrbuch der organ. Chem. 3rd Ed. p. 6.

2) Grundziige der wissenschaftl. Botanik I, p. 60 (1844)

3) Lezioni sui fenomeni fisico-chimici dei corpi viventi. 2d Ed. p. 10. (1846)

4) Kraft and Stoff, 3rd Ed. p, 256 (1856)

5) Lessons on elementary physiology, lesson VII (1870).

6) Generelle Morphologie der Organismen, Vol. I p. 143 and 148. See also:
Tageblatt der Naturforscherversammlung zu Munchen 1877 p. 22.

6 FORMER VIEWS ON THE CAUSE OF THE VITAL PHENOMENA.

force in the cells, he declared vital activity to be the result of
chemical processess whereby energy is liberated. What caused
those chemical processes he could not tell. Heidenhain takes it
for granted, that all the vital actions are nothing but peculiar
connections of physical and chemical forces exerted from the
living cells," and similar views were entertained by the well
known Chemical Physiologist of Great Britain: Halliburton,”
and by the physiologist F. Hiippe>) in Prague. It will be forever
remembered how Du Bois-Reymond undertook to draw bound-
aries of knowledge in pronouncing his ‘‘zgnorabimus”’ in regard
to the more complicated vital actions, at the meeting of the
German Association for the advancement of science, in Leipzig, 1872.
The learned professor however expressed 30 years earlier a
different view: ‘‘Those who preach the erroneous doctrine of
the vital force, under whatever form and in whatever disguise
it may be, such heads have—they may believe it—never
reached the boundaries of thinking.) ”’

The “ignorabimus” of Reymond would certainly not have
found favor with A. Humboldt, who in the year 1797 stigmatised
all exertions to draw boundaries of knowledge as paralysing
scientific activity.°) Of the German physiologists it was especially
Pfliiger who protested against such a thing by declaring: ‘* Who-
ever in our time undertakes to draw boundaries for the develop-
ment of science thinks his own brain-work worth at least just as
much as that of numberless generations to come.®)’’ We con-
clude with citing the view of the great French physiologist
Claude Bernard: “ Pour comprendre les fonctions de l’organisme
il faut connaitre celle de la cellule. La raison des phénomenes
vitaux est dans cette fonction élémentaire: le moyen de les
maitriser, de les modifier, d’agir sur eux, consiste a agir sur
activité cellulaire.” ”

1) Handbuch der Physiologie V p. 11. (1880).

2) Chemical Physiology, Chapt. 14.

3) On the cause of fermentations etc. Berlin 1893. p. 14.

4) Preface to the first edition of his work on animal electricity; p 39.

5) Versuche uber de gereizte Nerven-und Muskelfaser, Vol. II p. 77.

6) Die allgemeinen Lebenserscheinungen, Bonn 188s, p. It.

7) Lecons sur les phénoménes de la vie communs aux animaux et aux végétaux.
p. 458 (1878).

CHAPTER II:

MoDERN STEPS OF PROGRESS.

The attempts to penetrate the mysteries of the morpholo-
gical and chemical conditions of living cells are of comparatively
recent date. After Rk. Brown for the first time described the
cellular nucleus in the year 1833, and a few years later were
published the investigations of Schwann and of Schleiden on the
cellular structure of the organisms, when attention was drawn
by Dujardin to the apparent structureless, semifluid contractile
substance of some of the lowest kinds of animal forms, called by
him sarcode. Soon afterwards Mo/hl observed (1846) the pre-
sence of a similar substance in plant cells, which he called
protoplasm. Schulze demonstrated later the chemical identity and
showed that this substance lies at the base of all the phenomena
of animal and vegetable life. Every vital act depends upon the
some mode or property of protoplasm, which is of albuminous
character, and is the tangible reality, the chemical foundation
of life. That was the first important generalisation in the
domain of biological science.—Numerous observations followed :
the circulation in the vegetable protoplasm and its dependence
upon the access of air, the division of the nucleus and, quite
recently, the central corpuscles were discovered, the protoplas-
matic nature of the chlorophyllcorpuscles was demonstrated, the
leukoplasts were described. Important qualities of the cyto-
plasm became known and the different functions of the outer
and inner layer (cortical layer and tonoplast) were noted.

Attempts were even made to decipher the finer structure of
the cytoplasm and nucleus, and a reticular structure was dis-
tinguished from a hyaline and apparently uniform interfilar pro-
toplasm. It was further shown, that not a single chemical
process of importance can take place without the participation
of protoplasm; even each single starch-granule has its own
protoplasmatic manufacturer. Protoplasm is artist and tool
simultaneously, it cannot be a formless albuminous slime, it
must possess specific organisations according to the different
functions it performs. In the molecular condition of protoplasm

8 MODERN STEPS OF PROGRESS.

there is probably as much complexity as in the disposition of the
organs of the higher animals. The variations of protoplasm are
astonishing; the adaptivness to different functions unsurpassed
by any of the higher developed organisms. How different
must be the molecular arrangement in the celles of the bile-
producing liver from that of the cells of saliva-and again of the
cells of milk-producing glands! How different a machine
must exist in the cells of the resin-producing glands of the
conifers from that of the sugar-secreting cells of the nectaries.
What a wide difference again must exist between the organi-
sation of a fungus-spore and a pollen-cell! Only individualised,
specifically organised, till to the finest detail jointed and dif-
ferentiated structures can be the sources of the different actions
connected with life! Hanstein expressed this logical conclusion
in the following words: ‘‘ Even the most simple Protococcus or
Monad-cell is certainly of a complicated organisation, even if so
small that the best microscopes do not reveal a differentiation
of the cytoplasm; there is no vital action without a vital struc-
tuxe:-?
But nothing would be gained from the differentiation of the
protoplasmatic machines, if there were no motive power.
What kindles then the fire, that produces the necessary steam ?
That ‘‘ primum movens” must be united with the machine, most
intimately connected with it, and is absolutely necessary for
bringing on the most wonderful differentiations in form and
functions, which arise from a single original egg-cell, when
developing itself to an animal with the complicated system of
nerve,-muscles and gland-cells, with the different cellular forms
in epidermis and in bones. What a great variety of cellular
forms are found in a tree! How many cells must sacrifice their
individual existence during the development of an organism for
the formation of tubular systems and aquaeducts !

How in view of these most complicated phenomena the
older opinion can still be defended by some physiologists that the
protoplasm is a varying mixture of different substances, is a ‘ mix-
tum compositum’”’, remains incomprehensible to a logical mind.
Reinke found for instance in the plasmodium of Aethalium septicum
(a fungus belonging to the mixomycetes), 27 percent of carbonate

1) Das Protplasma, p. 308 ff, (1880).

MODERN STEPS OF PROGRESS. 9

of lime, and concludes, that this belongs to the molecular system
of this organism and contributes to its vitality! But if all the
substances found in protoplasm participated in the first cause
of life, then all excretionary substances that remain previous to
their expulsion for a certain time in the protoplasm, further the
sugar formed from starch and converted the next moment into
cellulose, the hydrocarbons, aldehydes and esters produced to
attract insects for fecundation of the flowers, the wax secreted to
form a film upon the epidermis of leaves, the urea and uric acid
produced in animals from proteids: all these substances would be
parts of protoplasm and would participate in the cause of vital
functions. All these combinations are formed in the protoplasm
of different cells and must exist there even if only for a short
time. This old view of declaring everything found in pro-
toplasm to be an essential part of the protoplasm, and attribut-
ing to it even vital actions leads simply to absurdities !—
Hanstein defined protoplasm “as a ‘semiliquid substance
consisting of proteids, insoluble in water, generally of neutral
reaction, transparent and refracting the light a little more than
water.”’ ‘This definition however is not perfect; we miss the
chemical side of the question, above all the distinction between
living and dead protoplasm. The first, who treated this problem
scientifically, was E. Pfliiger’) in the year 1875. He started
from the chemical fact that the living cells take up oxygen,
while the dead do not. From this fundamental difference the
conclusion is justified, that the albuminous substance of the
living protoplasm has another chemical character than that of the
dead protoplasm. In other words: the living protoplasm is ex-
ceedingly liable to chemical change, and if that change takes
place, death results. Pfliiger pointed out also, that the decom-
position of albuminous matter in the living animal organism
yields other products that the decompositions artificially accom-
plished in the laboratory and concluded, that the nitrogen of the
“living albumen” is linked in a different manner to other
atoms. He supposed that the cyanogen-group is this form,

1) Pfliig. Arch. Vol. 10.
2) This conclusion is however not justified; the same albuminous matter can

yield under different conditions very different products. The albumen, which is
oxydised in the living body is certainly for the greater part circulating dead albumen
and not living protoplasm. The latter brings on the oxydation of the former.

Io MODERN STEPS OF PROGRESS.

and that this form is changed by the fixation of water when the
cell dies :

ECN=+H,0=) CONE:
But it may be here objected, that such changes do not take
place spontaneously or easily.”

The conclusion however that a chemical difference exists
between living and dead protoplasm is plain logic.—If the dead
matter of our food is converted into the living matter of our
nerves, muscles and glands, a considerable chemical change
must take place and just in the opposite direction to that
connected with the loss of life. Pfliiger (I. c.) expressed his
conviction in the following words: ‘An albumen-molecule,
which in the brain concurs in the production of thought, which
in the spinal column mediates sensation, which in the muscles
performs mechanical work or in the glands starts chemical
activity, is doubtless derived from the same dead albumen of the
blood, but it is changed in chemical character as soon as it
enters the living cell.» From the moment it forms a part of the
living protoplasm, it commences to respire, to live. Only the
cells have the property of life; such albumen, which has not
become protoplasm, is dead albumen, even in the living body.”

The deductions of Pfliigey did not attract the attention they
deserved, they were even ignored by most physiologists. The
cause may be found on the one hand in want of knowledge of
the progress of modern chemistry, and on the other in the reac-
tionary situation created by the echo of the ‘‘ignorabimus” of
1872, still vibrating through the sultry atmosphere. Foremost
among those who assented, stood of M. Nenckt, who declared * :
‘‘T have repeatedly expressed my opinion, that investigation into
the albuminous bodies must take a new direction, if we want to

1) Pfliiger had here evidently the comparison with nitriles in view. Certain
other cyanogen-combinations, as cyanic acid, cyanamid, which undergo spontaneous
changes, of course can not serve here for comparison or explanation.—

2) This production of living matter from dead was declared by Pfliiger to be one
of the greatest enigmas of nature (Die allgemeinen Lebenserscheinungen, Bonn, 1889).
The supposition of Pfliiger that Liebig entertained the belief in a chemical difference
between albuminous matter in living and dead cells, is an error. Nowhere in
Liebig’s writings can a decisive opinion be found.

3) Arch. f. exper. Pathol. und Pharmacol. Vol. 20, p. 343. and Journ. f. prakt.
Chem. Vol. 26.

MODERN STEPS OF PROGRESS. rE

undertake with success the closer investigation of those actions
we call ‘‘life;”’ the proteids of the living cells must have another
chemical constitution than that of the dead cells.” And”:
‘It is of essential significance, that albuminous substances isolat-
ed at very low temperatures from living animals, are of a very
changeable character, as for instance the blood plasm, the oxy-
haemoglobin, the myosin.”’ “The most important function, nay
even the most characteristic feature of life itself is the formation
of labil albumen molecules.” ”

O. Rosenbach, discussing the vital phenomena® concludes:
*‘Death is the suspension of the labil equilibrium of atoms in
the living matter.” Among the plant-physiologists was Detmer
the only one, who defended the new ideas; but his modifications
will hardly find approval. He assumes, that the atoms of the
‘living albumen molecules” are in such a lively motion, that a
continuous dissociation of the albumen-molecules takes place,
forming thereby on the one hand non-nitrogenous substances as
glucose and on the other amides. By this dissociation respiration is
induced, and the other vital phenomena are made possible.) Such
a hypothesis however is incompatible with the great sensibility
of the protoplasm towards every disturbing influence. The con-
tinuous regeneration supposed by Detmer would be simply an
impossibility, as death would have resulted from the dissociation.
The view of Detmer would well nigh answer for a description of a
protoplasm committing suicide.

1) Ber. d. Deutschen Chem. Ges. Vol. 18, p, 385.

2) Pflugers Arch. Vol. 31, p. 336.

3) Aufgaben der Therapie, Chapt. 14. (1891).

4) Vergleichende Physiologie des Keimungsprocesses, Jena 1880,—Ber. d. Deut-
schen Botan. Ges, 1893.

CHAPTER, te

LIVING PROTOPLASM AND CHEMICAL LABILITY.

The view that at the moment of death a chemical change of
the living matter takes place, finds support not only in the loss
of affinity to the molecular oxygen of the air, but also in the
different behaviour towards certain coloring matters as anilin
colors, by which no living protoplasm, but only dead, can be
stained, the latter even if the coloring substance be very diluted.
Some phenomena accompanying the death of an animal are also
in this relation very noticeable, siz. the vise of temperature
observed, when soon after the death of the nervous system the
muscular tissue is also succumbing, not being nourrished any
longer by currents of oxygenated blood.

This socalled post mortal rise of temperature known long ago
and often incorrectly interpreted is really the mortal rise con-
nected with the proceeding death of the muscular tissue. Heat
is produced by chemical changes; chemical energy being easily
transformed into heat.

Intimately connected with the death of the muscles is the
formation, resp. the increase of an acid reaction and of rigidness.

All these facts can only be explained if the proteids of the
living cells undergo a chemical change; and we may therefore
distinguish them as active proteids from the passive proteids of
the dead protoplasm. The name “living albumen,” used some-
times, cannot signify anything else but living protoplasm, as life
is always the result of an organisation, of functions of proto-
plasmatic machines, and albumen itself without organisation
cannot be called “living,” even if a high lability should lead to
much chemical energy. All the steam of a locomotive would
not pull the train, if the machine were not properly constructed.
The name ‘‘ living albumen”’ should be discarded altogether, as
it might lead to erroneous conceptions. The name active
albumen, on the other hand, expresses merely a chemical
quality, a distinction of ordinary albuminous matter. — Still
preferable is the name ‘‘active proteids,” when the whole living
matter of a cell comes into consideration, because also active

LIVING PROTOPLASM AND CHEMICAL LABILITY. 13

nucleins and perhaps related proteids participate in the composi-
tion of the organoids of the cell. The active proteids become living
protoplasm by the process of organisation.

What kind of chemical character must then be ascribed to
the active proteids? We can answer this question but in one
and that at being decided sense: They are exceedingly labil com-
pounds that can be easily converted into relatively stable ones.
A great lability is the indispensable and necessary founda-
tion for the production of the various actions of the living pro-
toplasm, for the mode of motions that move the life-machinery.
There is a source of motion in the labil position of atoms in mole-
cules, a source that has hitherto not been taken into considera-
tion either by chemists, or by physicists.

The opinion that at a given temperature the motions of all
atoms in a compound are equal must be refuted as erroneous.
Labil atoms have a greater energy of motion than the stable
ones of the same substance.’) We know, that the atoms with
their sphere of oscillations occupy in different compounds
different volumes ;”) the oxygen in the aldehydegroup occupies a
larger volume than in the hydroxylgroup, the ratio being 1:0, 6.
The atomic volume of nitrogen in the cyanogengroup is larger
than in the nitrogroup and here again larger than in the amido-
group, the ratio being 1:0,50:0,13. Andif the atomic volume
of hydrogen were determined carefully in different combina-
tions we should very probably find, that it occupies, in an
aldehyde-and in an amidogroup a larger volume than in a
methyl-and in a hydroxylgroup.

It will perhaps be not out of place to explain the nature of
chemical lability in a few words. .

A labil position exists, if in a molecule one atom is influ-
enced simultaneously by the affinities of two neighboring atoms.
Thus lively oscillations are produced bringing on a great ability
for reactions and an inclination for a spontaneous migration of
the labil atom into a stable position.

1) Atoms moved simultaneously by heat and by chemical affinity must possess
more energy than those moved by heat alone.

2) Unsaturated carbon compounds occupy a larger molecular volume than
calculated from the numbers belonging to the saturated ones.—Aldehyde has further
a larger specif. volume than the isomeric ethylenoxyde, the ratio being 56, 4: 50, 6.

I4 LIVING PROTOPLASM AND

One of the most interesting labil atomic groups is the
aldehydegroup Ost in which the oxygen exerts an attract-
ing influence upon the hydrogen connected with the carbonatom,
which is generally tetravalent, but can in some instances also
functionate bivalent. Thus the hydrogenatom is subjected to
continuous vibrations from the carbon to the oxygen and back,
as may be indicated by the following formulas:

aces BH oa =052 =¢

(1) (2) (3) (4)

These motions are accelerated by rise of temperature ; in the

—-O-—H

same measure the inclinations to chemical reactions increase.
Ammonia, diamid, hydroxylamin, hydrocyanic acid, sulfuretted
hydrogen, primary sulfites act with great facility upon alde-
hydes and even the molecular oxygen is taken up easily by
certain aldehydes. Certain substances bring on a rapid change:
thus a little sulfuric acid will transform ethylaldehyde into
paraldehyde, a polymeric modification, whereby a contraction
and development of heat takes place. Caustic potash converts
that aldehyde into a resin.

Another group of a certain lability is represented by the
following position :

—C=0 —C—OH
| easily passing into: I
—CH, —CH

One of the laws of lability can therefore be generally ex-
pressed thus: Jf in a chain of carbonatoms one of these atoms has
two affinities saturated by one oxygenatom, while the other two
affinities of the same carbonatom are saturated hy positive atoms or
groups of atoms, a labil group is formed. This lability is increased
with the entrance of stronger positive groups and is lessened by
negative groups ; amido-benzaldehydes show a far greater lability
as nitvo-or oxy-benzaldehydes.

The saturated hydrocarbons, alcohols and acids of the
methanseries are in comparison with the aldehydes very stable
compounds; also the saturated hydrocarbons of the benzol
series. The stability however decreases with the number of
entering hydroxylgroups.

Now taking the proteids into consideration, it must be borne

CHEMICAL LABILITY. 15

in mind, that these are the most complicated combinations of
all, and that up to the present day our slow inductive methods
have not yet elucidated their chemical structure, although
numerous facts relating to the decompositions under different
circumstances have been discovered within the last ten years,
especially by Maly, Nencki, Drechsel, E. Schulze. It was there-
fore not possible to look here for an explanation in regard to the
constitution of the active albumen or proteids. A more pro-
mising way seemed to me to study at first the formation of
albuminous substances within the life-plant, as it was most
-probable, that the active proteids were products of direct
synthesis. In this regard an important observation made at
first by Th. Hartig and later studied by Borodin, Pfeffer, Kellner
and especially by EZ. Schulze put us upon the right track: the
observation that in many cases of albuminous decompositions in
plants asparagin is formed in surprisingly large quantities, and
that this asparagin quickly disappears again in the formation
of new proteids, without any intermediate product being dis-
cernible. (See Chapt. V.).

This led me to the conclusion, that albumen is formed by
a rapidly proceeding condensation-process. But in order to
render this possible, the asparagin had to be transformed first
into an aldehyde, the aldehyde of aspartic acid, from which
by condensation and reduction substances of the composition of
albuminous matter could be formed, as may be expressed by the
following equations :

I. 3(C,H,NO,)=C,,H,,N;0,+2H,O

II. 6(C,,H,,N,0,)+12H+H,S=C,,H, ,.N:3SO,,+2H,O0

Simplest expression for albumen

If the condensation were to take place between the alde-
hyde-and methylengroups and the hydrogen indicated in the
equation II were to be used for reduction of 12 aldehydegroups,
the final product would still contain r2 aldehyde-and 18 amidogroups
and would therefore be of an extraordinary lability. The
energetic motions in such a product could bring on oxydative
processes (see Chapt. VI.), in which easily oxydisable products
as sugar, lecithin, could participate. This process might lead in

1) O. Loew. Pfliig. Arch, Vol. 22, p. 503.

16 LIVING PROTOPLASM AND

the living protoplasm by regulating contrivances to the respira-
tionprocess.

The question arises now: can it be proved directly or
indirectly that aldehyde-and amidogroups are really present in
the living protoplasm ?—If our hypothesis is correct, that the
vital motion emanates from labil aldehyde-and amidogroups, it
follows, that every substance reacting in great dilution with
those groups must prove a poison for every living vegetable or
animal organism.”

As experiments confirmed this conclusion, our hypothesis of the
formation and nature of active albumen has thus received essential
support. Very correct views were expressed by Nencki in regard
to the cause of poisonous actions.» ‘‘ Why is it that a poison-
ous substance has a well defined action, while slight changes in
the chemical constitution of the poison bring on quite different
actions? This depends on the one hand upon the chemical
constitution of the protoplasm and on the other hand upon the
chemical structure of the poisonous substance.” Indeed in most
cases the poisonous actions are the result of a chemical reaction
upon the albuminous substances of the living cells. If we lessen
the chemical energy of a poisonous substance by introducing a
carboxyl-,-or a sulfogroup, the physiological action will be also
lessened. If we substitute alkyls for the hydrogenatoms of
amido-or imidogroups the poisonous character is decreased ;
while on the other hand innocuous substances might turn into
poisons if imidogroups, or amidogroups of a certain lability are
formed. Thus the non-poisonous hydrobenzamid can be trans-
formed by atomic migrations into the potsonous amarin, isomeric
to the former.—

And what an enormous change in the poisonous qualities is
observed, if one hydrogenatom of the ammonia-molecul is re-
placed by the hydroxyl,-or by the amido-group! While ammonia
in form of neutral salts is in a certain dilution no poison for
animals, the substitution-products thus obtained, viz. the hy-
droxylamin resp. diamid in the same dilution in neutral solutions
are very strong poisons.—Bacteria and mouldfungi, that are not

1) Compare O. Loew, A natural system of poisonous actions, Munich, 1893.
Chapt. 4.
2) Arch des scienc. biolog. St. Pétersburg 1892 p. 61.

CHEMICAL LABILITY. 17

poisoned by concentrated solutions of neutral ammoniasalts, are
easily attacked by hydroxylamin even at dilutions of 1: 10,000").

I found, that diatoms are killed within 24 hours by hydroxyl-
amin in a dilution of 1: 100,000 and that in a dilution of 1: 20,000
it is a stronger poison for infusoria than strychnin! Ina dilution
of 1: 15,000 it kills within a few days phaenogamous plants, as
was shown by me with young plants of maize and helianthus and
by E. Schulze with maize and barley.—Marpman observed later
also the poisonous qualities for pathogenic microbes. In short,
while ammonia is next to nitrates the most important source
of nitrogen for the nutrition of plants, hydroxylamin never can
be used for this purpose, being a deadly poison !—

In regard to lower animals I observed, that while sal-am-
moniac in a dilution of 1:10,000 had no noxious effect upon
crustaceans (copepodes) the equally diluted solution of hydro-
chlorate of hydroxylamin (neutralised with soda) killed them
within 3 hours. A solution of 1:20,000 killed aquatic snails,
worms, and larvae of insects within 36 hours.—Raimundi and
Bertom, Binz, Lewin studied the poisonous action on the higher
animals. 0,05g. of hydrochlorate of hydroxylamin kills a pigeon
in 3 minutes; 0,1g pro Kilo. of a warmblooded animal will pro-
duce convulsions.

When the diamid was discovered by Th Curtius in the year
1887, and its remarkable affinities for aldehydes became known,”?
I predicted at once, that this substance must prove a universal
poison, a poison for all kinds of living cells. Indeed I observed
afterwards that the neutralised sulfate in a dilution of 1: 10,000
kills algae in 1-2 days; at 1: 5,000 bacteria ; at 1: 2,000 infusoria,
crustacea, mollusca and aquatic larvae of insects within 12 hours,
while neutral ammoniumsulfate in this dilution produced not
the slightest effect whatsoever. I found further, that young plants
of Helianthus died within 4 days, and of barley within 2 days,
if placed into a solution containing 0,2 p. mille neutralised
sulfate of diamid, while the controlplants treated with equal
quantities of ammoniumsulfate developed normally.*) 0,5 ¢. of

1) O. Loew, Pfliigers Arch. Vol. 35, p. 515 (1885).

2) According to Curtius diamid attacks aldehydes even in strong acid solutions,
while it combines with ketons only in form of the free base.

3) O. Loew, Berichte d. D. Chem. Ges. Vol. 23, p. 3204.

18 LIVING PROTOPLASM AND

diamid sulfate will kill a rabbit in 1} hour (Buchner).

Why is it then,—it will be asked that hydroxylamin and
diamid are such strong poisons compared with ammonia? The
answer can only be found in the marked difference of behaviour
to aldehydes (resp. Ketons). It has been demonstrated by
recent investigations, that salts of hydroxylamin and of diamid
(hydrazin) act even in high dilutions upon aldehydes while
ammonia acts only as free base or as carbonate and with less
energy. This difference is due to the greater lability of the
hydrogenatoms in the former, attacking thus easier the labil
oxygen of aldehydes and ketons.

ae ve we
Not N—H N—H
\ H BS OH \ NHa2
Ammonia. Hydroxylamin. Diamid.

We find analogous differences of intensity of toxicological
actions between anilin C;H,.NH, and phenylhydrazin, C.H..
NH.NH,; between pyridin C,H,N and piperidin C,H,,.NH;
between phenol and amidophenol, which latter is,-im accordance
with the behaviour to aldehydes,-also a stronger poison than
anilin.—

The reaction between hydroxylamin and an aldehyde is
expressed by the following equation :

= =—N—OH
(@) Ci, +HIN= On = @)=C2ne eee
——— ee
An aldehyde. An aldoxim.

It is of considerable interest, that also certain derivatives
of hydroxylamin react easily with aldehydes and exhibit in ac-
cordance therewith a poisonous character; these derivatives are
the ‘‘amidoxims,” prepared by Tiemann by the action of con-
centrated hydroxylamin upon nitriles at higher temperatures :

(@)+C=N+H,NA0H = @e Onna
A nitrile) An amidoxim.

()— Cin on @)—Can, =) Cm -cErons

————— ~
An aldehyde.

1) The product thus formed can undergo an atomic migration; Tiemann,
Ber. 22, p. 3124.

CHEMICAL LABILITY. Ig

One of the amidoxims, the benzenylamidoxim

C,H, “estes was shown by Mering to be poisonous:
0,5 g. will kill a dog; 0,1 g. a rabbit ; 0,03 g. a frog.”

But we observe also on the other hand that substances
acting readily upon labil amidogroups are equally poisonous;
above all may be mentioned the formic aldehyde, the free
cyanogen, and the nitrous acid, which latter is far more poison-
ous than nitric acid. I observed that formic aldehyde CH,O in
a dilution of r: 10000 kills readily microbes and algae; of I: 2000
in 2 hours crustaceans, worms and mollusca.*”) Rabbits are killed
by 0,24g. pro Kilo. (Zuntz). Phaenogams watered with a I p.
mille solution will die in from 2-6 days (Bokorny). This aldehyde
annihilates also as I have found (Jahresber. f. Thierchemie 1888)
the actions of the enzymes. The action on amines is shown by
the equation :

H,CO+H,N—(x) = H,C=N—(x)+H,0

The free cyanogen is for lower vegetable and animal organ-
isms a stronger poison than hydrocyanic acid; in a dilution of
1: 5000 it kills cells of the bear-yeast, in dilution of 1: 10000
infusoria and algae.) Its action upon labil amidogroups may be
represented by the following equation:

=NH —NHz
CN C _NnH-(z) acta

CN CN CN

The action of mztrous acid upon labil amidogroups is shown
by the following equation:

NO OH+H,N—(«s) = OH—N=N—(x)+H,0
and:
OH—N=N-—-(s) = OH—-(«)+N,

Free nitrous acid in a dilution of 1:100000 kills algae
within 48 hours (Loew and Bokorny*)). Nitrites are poisonous
in all those cases, where nitrous acid is set free by organisms,
thus for plants containing free acids or acid salts, and for the

1) Ber. D. Chem. Ges. 1885 p. 1054.

2) The great antiseptic properties of formic aldehyde were first observed by
myself. Jahresb. f. Thierchemie 1888.

3) According to investigations which were carried on by myself and Mr. Tsuka-
moto in the laboratory of Komaba.

4) Botan. Zeitg. Dec. 1887.

20 LIVING PROTOPLASM AND

higher animals. 0,03g. sodium nitrite kills a frog, 005g a
rabbit (Emmerich and Tsubot:;’) Binz).

We observe therefore: The conclusion that the labil atom-
groups in the living protoplasm are aldehyde-and amidogroups,
agrees exactly with physiological phenomena, with toxicological
facts, which cannot be said of the views of Pfliiger and of
Latham assuming the labil groups to be cyanogengroups. Ac-
cording to the view of Pfliiger the chemical change connected
with the loss of life consists in the chemical fixation of the
elements of water, according to our view, however, in a migration
of atoms from labil to stable position without water being taken
up. Cyanides will react with hydroxylamin only in concentrated
solutions and at higher temperatures, aldehydes however in the
cold and in high dilutions. The poisonous actions of all
the substances above mentioned, furthermore, are not in ac-
cordance with the cyanogen-hypothesis.

As labil amidogroups act readily upon labil aldehydegroups
there arises a constant danger to the living cells themselves of
an inter-molecular poisoning taking place; indeed we need only
to heat the living organisms to 45-50° and death will result with
rare exceptions; the continuous danger has become a reality.
I expressed this chemical change by the following equation:

—CH—NH, —CH—NH
Wore = pte,
———
An active group2). A passive group.

Substances that contain simultaneously aldehyde- (resp.
keton-) groups and amidogroups are indeed of an extraordinary
lability and when I first published my views upon the formation
of active albumen in plants (1880, Pfliig. Archiv.); no such
combinations were yet known. But soon afterwards the or-
thoamidobenzaldehyde was prepared by Friedlaendey and found
to be a very reactive substance.

1) These two bacteriologists have demonstrated that also the poisonous symp-
toms in cholera depend upon the formation of nitrites from nitrates by the cholera-
bacillus.

2) Twelve of such groups are—according to my hypothesis of the formation of
active albumen—contained in one molecule of the latter; if the simplest expression
for albumen is taken as a foundation.

CHEMICAL LABILITY. aI

Later followed the preparation of paraamidobenzaldehyde
and of amidovaleraldehyde.’) Quite recently the amidoethyl-
aldehyde was prepared by E. Fischer.?) This substance is only
stable in form of salts, and changes so rapidly after being set
free, that it loses its power of reducing Fehling’s solution within
one hour, and becomes transformed into a gelatinous substance.
Also the diamido-aceton soon changes spontaneously to an amor-
phous substance if set free from its combination with acids.°
We observe therefore, that there exist labil amido-combinations
which undergo spontaneously a rapid change,—losing thereby
their original characteristics.—That here is a certain analogy to
the change of protoplasm, when it passes from life to death, can
hardly be denied if we see, that hydroxylamin and diamid, the
most characteristic properties of which is their great ability to
react in high dilution and at ordinary temperature upon al-
dehydes,—have no longer any action upon dead protoplasm, nor any
influence upon ordinary soluble proteids at ordinary temperature.”
—Those physiologists, who still cling to the old notions of the
identity of proteids in the living and the dead protoplasm, will
never have an understanding of the cause of the vital functions,
they will be at a loss to conceive the method of poisonous
actions !

If a change takes place from a labil to a stable character,
if the cell dies, the reverse transformation must succeed, if by
growing and multiplying animal cells pepton is resorbed for the
formation of living protoplasm. Here kinetic energy is trans-
formed into potential energy, passive groups become active ones.
Labil combinations occupy a larger molecular volume than the
isomeric stable ones, hence work must be produced by the
conversion of the latter into the former. On the other hand
contraction and development of heat will result if the active,
labil albumen passes into the passive condition. Contraction
of the protoplasm is indeed taking place with the death of the

1) Wolffenstein, Ber. Deutsch. Chem. Ges. Vol. 25, p. 2777.

2) Ibid. Vol. 26, p. 92.

3) Riigheimery and Mischel, Ibid. Vol. 25, p. 1563.

4) It would be incompatible with the spirit of science to avoid the logical
conclusions to which the toxicological facts mentioned, must lead. There exist
physiologists who would declare all conclusions premature, but have they considered
what good grounds exist for such an assertion ?

22 LIVING PROTOPLASM AND CHEMICAL LABILITY.

cells"), and the heat developed on the death of larger cellular
complexes like muscles can be measured with the thermometer.—

The vital phenomena depend upon the labil aldehydic charac-
ter of the living protoplasm; this labil character gives rise to
respiration. Respiration produces heat, and heat again increases
the oscillations of the labil atomic groups in the living proto-
plasm, until a certain limit is reached. Hereby a transformation
of heat into chemical activity results, the vital motion thus
leads to vital functions. We propose to call the resulting vital
motion: plasmic force, instead of vital force, which name would
recall the erroneous conceptions of former times. We define
therefore the plasmic energy as a mode of motion produced by ‘the
labil atomgroups in the proteids of the living protoplasm and intensifi-
ed by the process of respiration, induced by the labil character.

However not only is the chemical structure of the proteids a
labil one, but so also is the morphological structure of the proto-
plasm, the tectonic, i.e. the invisible arrangement of the molecules
of active albumen to small particles of protoplasm, composing
the organoids of the cells (nucleus, chloroplasts, filarplasm,
tonoplast etc.).7) Slight disturbances of a mechanical nature
produce contraction and death. The chemical attack of a poison
upon only a mznute portion of the protoplasm of a living cell may
cause the collaps of the entire protoplasm. The morphological
construction and the chemical nature of the living protoplasm are
evidently most intimately connected; an injury to one will also
damage the other: The living protoplasm is a labil structure built
up of labil material.

1) Contraction of cytoplasm in plantcells is not observed however in some
special cases, as treatment with absolute alcohol, acids and dilute caustic lyes. But
an invisible contraction has nevertheless taken place also in these cases, as can be
demonstrated by the loss of the osmotic qualities of the cytoplasm. The pores have
become so large, that the osmotic membrane has changed into a filter, that now
permits easily the exit from the vacuole of various substances, as tannin etc.

2) It is convenient to distinguish the invisible organisation as ‘‘ tectonic”’ from
the visible organisation, i.e. differentiation into different organoids of a cell.

CHAP Thi tv.

ACTIVE ALBUMEN AS RESERVE-MATERIAL IN PLANTS.

The facts described in the last chapter lead us immediately
to the questions: Do the active proteids also exist as such, 1. e.
in the xon-organised condition, in plantcells? If so, are they just
as labil as in the organised state, i. e. as living protoplasm?
Is it possible to obtain directly the chemical reactions upon
the aldehydnature of those compounds ?—To these questions
Dr. Thomas Bokorny and myself have devoted considerable time
and attention and our investigations’) have shown: 1. That
there exists in many plants, apparently in a state of solution a
certain protein-substance quite different from ordinary proteids ;
2. That this substance is capable of giving certain reactions, of
which the living protoplasm on account of its great lability is
incapable, and which neither dead protoplasm nor the known
soluble proteids show; 3. That this substance is used up
during the growth and multiplication of cells, and that it plays
therefore the réle of a reserve-material.

The reader will be made acquainted in the following lines
with the principal observations relating to this discovery.77—
Many vegetable objects, algae as well as parts of higher deve-
loped plants, show under the influence of weak bases like coffein
(0,1-0,5%) or antipyrin (best in 0,5% solutions) a remarkable
phenomenon, consisting in the appearance of a large number
of minute transparent colorless globules,*?) that gradually unite

1) Of our publications I mention here merely:

O. Loew and Th. Bokorny: Die chemische Kraftquelle im lebenden Pro-
toplasma, Munich 1882.—
Botan. Centralbl, 1889, and 1893.—Biol. Centralbl. Vol 11.—
Flora 1892. p. 117.—

Dr. Th, Bokoryny, now professor at the Military Academy in Munich, publish-
ed contributions in:

Pringsheims Jahrb. Vol 19. and Vol. 20. Pfliig. Arch. Vol. 45 and Vol. 50.

2) I describe here the facts in the order, in which it appears most convenient
for the reader to conceive. I neither enter therefore upon a sketch of the develop-
ment of our investigations, nor upon a relation of the discussions brought on by
our publications. Suffice it to say that the objections have been refuted as wholly
unfounded.

3) Coffein acts still in higher dilutions than antipyrin.

24 ACTIVE ALBUMEN

into larger globules and droplets, losing thereby their original
motions (Brown’s so-called molecular motions?). As regards
most of the objects these droplets are situated in the vacuole, in
some however in the cytoplasm as well. All kinds of Spirogyra,
an alga of common occurrence, are for these observations es-
pecially well adapted; they remain for some time—even for a
series of days—alive in the solutions mentioned. If the objects,
in which the droplets have been produced, are taken from these
solutions and replaced in pure water, the droplets will gradually
disappear again in proportion as the bases mentioned leave the
cells by osmosis. The cells continue thereby their life as before
the treatment. This dissolving process is quickened at higher
temperatures, at 30°C. it requires but a few minutes. Replace-
ment of the objects in the solutions of these bases makes the
droplets reappear. If however the cells die by the prolonged
influence of coffein or antipyrin, or if they are killed by dilute
acids or by poisons like formic aldehyde, hydroxylamin, or salts
of copper, etc., or by vapors of ether, then these droplets also
change their properties, soon after the death of the cells. Hereby
the close chemical resemblance of matter in the protoplasm and in
the droplets becomes manifest. The droplets become turbid from
numerous and minute vacuoles, formed by a sudden loss of ab-
sorbed water, and, in coagulating, lose their solubility.) In some
cases the small vacuoles unite into one large one, a hollow sphere
thus resulting, in other cases the spherical form is lost entirely,
leaving an irregular shaped mass. In some objects the vacuoles
disappear again by. further contraction, and the globules again
turn transparent; the insolubility however remains. This turn-
ing from the soluble into the insoluble state is a decisive proof of
a chemical-change.—

It is of special interest to observe under the microscope the
change brought on by the action of a 0,2-0,5 per cent solution of
acetic or sulphuric acid or by a diluted alcohol of 10-20 per cent.”

t) It is of rare occurrence, for the droplets to lose their solubility, before the
death of the cells is observed in the above named solution of coffein or antipyrin.

2) If instead of diluted alcohol absolute alcohol is employed, the coffein is so
rapidly extracted that the smaller globules dissolve before they are coagulated, while
the larger ones shrink to irregular shaped thin films. Here the exosmose of coffein
proceeds quicker, than the endosmose of a sufficient quantity of alcohol. All the
experiments mentioned are best made with the larger globules ; the changes cannot
be well observed if the globules are too minute.—

AS RESERVE-MATERIAL IN PLANTS. 25

Still more striking is the effect of the vapors of ether. If
spirogyra-threads containing freshly produced droplets are ex-
posed in a flask at ordinary temperature to the vapors of ether,
the cells are found killed in a few seconds, and about 20 minutes
afterwards: all the globules change their aspect, losing their
brightness and their solubility!

The coagulation by heat is easily observed if the objects are
dipped in boiling water containing 5 per cent of chloride of
sodium, all droplets exhibiting then a turbid appearance ; neither
boiling water nor absolute alcohol will change them any fur-
ther. It is well known, that the presence of salts facilitates the
coagulation of albuminous matters.

The substance in question is also changed quickly in the
dissolved state, with the death of the cells; in dead cells coffein
never produces any globules. If for instance we treat Spirogyra
Webert for one minute with a very dilute aqueous solution of
iodine, the globules may be still produced by coffein immediately
afterwards, but after 10 minutes action of the iodine, no more.
That our substance had not left the dead cell by osmosis can
be easily shown, if we add to the small quantity of the iodine-
solution, in which we left a larger quantity of the algae to die,
coffein in a sufficient quantity: no trace of the above described
phenomenon is observed. The same experiment may be made
with cells killed in any other way.

All these facts demonstrate beyond a doubt, that we have a
peculiar protein-substance before us, distinguished from the or-
dinary soluble proteids not only by its being separable from the
dissolved state in globules by coffein or antipyrin, but also by a
very great lability, as these globules become coagulated by
influences, which do not at all change the ordinary soluble pro-
teids, as alcohol of 20 per cent or vapors of ether. We called
these globules protcosomes, thus reminding us on the one hand
of the proteids, on the other of the ‘‘microsomes.” In the
coagulated state they exhibit all the properties of ordinary co-
agulated proteids. Treated with phosphortungstic acid (10 per
cent) they remain insoluble even after weeks, while hydrochloric
acid of to per cent changes them gradually and dissolves them
after a series of days at ordinary temperature.—A solution of
“caustic soda or potash of moderate strength soon dissolves the

26 ACTIVE ALBUMEN

proteosomes, but if they had been treated with a neutral solution
of formic aldehyde (5-10 per cent), they would have lost their easy
solubility in caustic lyes,—exactly like the behaviour of ordinary
proteids, I observed some time ago...—The Millons reaction is
obtained if the objects are left for 8-10 hours in a solution of
mercuric nitrate containing some potassium nitrite, and then
heated for a short time to the boiling point.—The ‘‘biuret-
reaction’? is obtained on treating the protecsomes first with
diluted ammonia (0,1 %), then for 12 hours with a dilute solution
of acetate of copper, and finally with dilute caustic potash.?)—
While the fresh proteosomes soon disappear by treatment with
dilute carbonate of soda, such proteosomes, as were changed by
the death of the cells, offer considerable resistance to it.

Of special interest is the behaviour to diluted ammonia,
whereby the proteosomes are solidified, another and marked dif-
ference from ordinary proteids being thus demonstrated.*) These
solidified proteosomes are chemically not identical with the co-
agulated proteosomes mentioned before; the ammonia brought on
a different change, being taken up and having entered into an
intimate combination, which however has no salt-like constitution,
as the chemical behaviour clearly indicates. A dilute hydro-
chloric acid (0,5 per cent) will at 40°C gradually attack the
coagulated proteosomes, but not those solidified by ammonia.
Even a to per cent hydrochloric acid dissolves at 80° the latter
ones with difficulty, compared with the former. This chemical
fixation of ammonia recalls the formation of pyrrols from 1.4
diketons or the formation of aldehyde-ammonia-groups, with
subsequent changes to a more intimate fixation of the nitrogen.
If aldehyde-groups be present in our labil proteosomes, the
behaviour towards ammonia can be understood, and as aldehydes
retain their silver-reducing properties even after combining with
ammonia, this question could be settled by experiment. But
care had to be taken to exclude every trace of other reducing
compounds as glucose or tannin, frequently occurring in plant-
cells.—

1) Compare O. Loew, Jahresbericht f. Thierchemie 1892 p. 29 and 1888 p. 273.

2) For further details see Botan. Centralbl. 1889 Nr. 39 and 45.
3) This behaviour to ammonia may serve in some cases to distinguish small and

indistinct proteosomes from tannates of coffein or antipyrin, which will be readily
dissolved by ammonia.

AS RESERVE-MATERIAL IN PLANTS. 27

To select objects free of glucose, is not difficult, but rarer
are such as unite with an abundance of proteids the absence of
tannin, seemingly a frequent by-product in the synthesis of
proteids. It collects in the proteosomes in combination with
albumen.—We have shown however, that tannin might be used
as a source of carbon in the formation of proteids under favorable
circumstances. These experiments were made with Penicillium,
cultivated in nutrifying solutions containing as sole organic sub-
stance tannin. The produced mass of the fungus amounted to
12,4 per cent of the tannin applied, after a growth of four weeks,
at the end of which time no trace of tannin was present any
longer.’) This result led us to experiments with Spivogyra” and
indeed the tannin disappeared, the best reactions failed finally,
and when the cells were boiled with a little water, neither the
acqueous liquid nor the cells themselves showed any reaction
with a 1 per cent solution of nitrate of silver supersaturated
with ammonia; hence it was evident that neither a soluble
nor an insoluble reducing substance was present any longer
in the killed cells. Still, the proteosomes were capable, even
after treatment with ammonia (1 p. m.), of reducing even very
highly diluted alkaline silver solutions, the globules thereby turning
black. Such proteosomes however as were changed by acetic
acid, or by any other death of the cells were found incapable
of reducing the same silversolutions, even after 24 hours contact.
This silver reagent was always left with the objects in the dark,
and was applied ina highly diluted state, because it is charac-
teristic of aldehydegroups, to bring on a silver reduction in far
greater dilutions, than many other reducing substances or atomic
groups. A suitable silver solution may be obtained by super-
saturating 1 cc. of a I per cent solution of silver-nitrate with
ammonia, adding a few drops of a diluted solution of caustic
potassa and diluting this mixture to one liter with distilled
water. This solution contains 100000 parts of water for 1 part
of silver-nitrate employed (present now as oxyde in solution) and
is still easily affected by aldehydes.

1) Botan. Centralbl. 1889 Nr. 39.

2) The smaller kinds of Spirogyra are sometimes found also in nature free
of tannin. Of the larger kinds most tannin is found in Sp. crassa and Sp. majuscula,
which therefore are not so favorable for these experiments, as other kinds.

iS)
2)

ACTIVE ALBUMEN

The solution, in which we cultivated Spirogyra nitida and
Sp. Weberi for the purpose of making the tannin disappear,
contained: calcium-nitrate, magnesium-nitrate and potassium-
sulphate, 0,05 per cent of each, monopotassium-phosphate 0,005
per cent and a trace of chloride of iron. A few threads were
placed in a liter of this solution, which was at a temperature
of 15—16° left exposed to only a moderate amount of light for
2—3 weeks.”

An object entirely free of tannin or related compounds are
the snowberries (Symphoricarpus racemosus). If the fleshy tissue
of unripe snowberries be treated first with coffein, and the pro-
teosomes be left in diluted (0,1 per cent) ammonia, the tissue
then washed with tepid water to remove every trace of sugar, we
observe also here an intense blackening of the proteosomes by
the silver reagent mentioned.

The question, whether our labil proteosomes represent the
active albumen of our theory is easily proven. If favorable con-
ditions for growth and multiplication of cells were offered and at
the same time the formation of new albumen was made im-
possible, then the reserve-albumen had to be used up gradually.
We shall indeed observe this result if we cultivate for instance
threads of Spirogyra in a comparatively large volume of the
following solution :

0,05 per cent calcium sulfate,

0,02 ,, ,, calcium bicarbonate,

0,02 ,, ,, magnesium sulphate,

0,005 ,, ,, monopotassium phosphate,

Small trace of chloride of iron.

In this solution were present all the mineral constituents
necessary for development, except every suitable source of
nitrogen, hence new albumen could not be formed and the
reserve albumen had to be seized upon for the organisation of
growing protoplasm, for the nucleus and chlorophyll-bodies of
the new cells. Indeed the stored-up albumen disappears here
so perfectly, that coffein fails to produce proteosomes after 2-3

1) Hereby the quantity of originally present active albumen is found relatively
decreased in the cells, on account of their multiplication, but the proteosomes produced
remain perfectly colorless, if the threads are left to die in a solution of ferrous
sulphate ; a proof, that every trace of tannin was used up.

AS RESERVE-MATERIAL IN PLANTS. 29

weeks of cultivation. The albumen of our proteosomes therefore
serves for building up protoplasm.—-If we place now the same
threads in a solution containing fotassiwm-nitrate, as for in-
stance in:

0,05 per cent potassium nitrate,

O03" 5, ., Calcium nitrate,

0,005 ,, 5, magnesium sulphate,

0,005 ,, ,, monopotassium phosphate,

trace chloride of iron,
we obtain after 3 weeks with coffein an intense formation of
proteosomes, more active albumen having been produced than
was required for organisatory purposes. Potassium nitrate is
not only a suitable source of nitrogen, but the potassium of this
salt is also important for the chlorophyll function, yielding an
increased quantity of starch for the synthesis of proteids. Spzro-
gyra majuscula forms in this solution after several weeks such
large quantities of active albumen, that this commences in some
cells to separate in globules even without the coffein-treatment.—

Also changes of temperature have a great influence. Hot
weather favors growth and the active albumen may be more
quickly used than formed; hence a decrease of the reserve-
albumen; if now cold weather follows, growth is more retarded
than the synthetical preparation of proteids, hence an accumula-
tion of active albumen results...—Also phosphates interfere
with the accumulation. In the absence of these salts (other
conditions being favorable) albumen will continue to be formed,
but organisation and multiplication of cells will have stopped,
hence accumulation increases.”

1) It was quite natural for us to take, at the beginning of our studies, the
proteosomes of the cytoplasm for an essential part of the protoplasm, as it was not
known before, that dissolved proteids occur as reserve material in the living pro-
toplasm itself. The recent attack of P. Klemm upon our definition of active albumen
(Bot. C. March 1894) contains no arguments, but merely a series of assumptions
disregarding the main facts we described, especially the most important change
of the proteosomes as soon as the cells are killed in one way or other.

The state of copulation does not depend upon the amount of stored up active
albumen; I have observed in some cases much of the latter, in other cases none
at all,.—

2) O. Loew, On the physiological functions of phosphoric acid; Biolog. Cent-
tralbl. Vol 11. p. 280.

30 ACTIVE ALBUMEN

Active albumen can be stored up in higher and lower plant-
forms and in most different parts of plants. Thus we obtain
proteosomes by coffein or antipyrin with leaves of all insectivo-
rous plants except with Utricularia. Drosera shows it in all parts
of the leaves, stem, and flower.*? The most remarkable however
among all insectivorous plants is in this regard Cephalotus, con-
taining in all parts astonishing quantities of stored-up active
albumen. Further objects are: The subepidermial cells of the
leaves of Cvrassulaceae as Cotyledon, Echeveria, Sedum; the
epidermial cells of Primula sinensis and Pelargonium; the hairs
of the stems of Begonia and of many other plants; the petals of
Cyclamen, Cornus, Tulipa, Epidendron; the anthers of Eugenia,
Melaleuca and Forsythia; the pistils of Crocus vernus, Rhodo-
dendron, Salix, Euphorbia; the peduncles and petals (sometimes
also young seeds) of:

Gentiana, Primula, Scrofularia, Impatiens Sultani,

Hoteia japonica, Pirus Malus, Prunus Cerasus,

Amorphophallus Rivieri, Viburnum rugosum,

Sorbus aucuparia, Thea chinensis ;
the flowers and young leaves of Rheum, Acer, Populus, Acacia,
Crataegus oxyacantha, Mimosa pudica ; the nectaria of Passiflora.

In some plants it is found only at certain stages of develop-
ment, as in the petals of malvae, in unripe snowberries (Sym-
phoricarpus racemosus), in cotyledons of Helianthus, in the
larger cells of Vallisneria-leaves.

Whether fungi ever store up active albumen as a reserve-
material is still doubtful;? among the algae are the various
kinds of Spirogyra the most noticeable ones, Vaucheria contains
it occasionally, Mesocarpus, Oscillaria, Oedogonium, Sphaero-
plea, Palmella, Desmidiaceae and Diatomeae never, so far as
our observations reach. Of higher organised plants not contain-
ing this reserve-material may be mentioned: Leaves and roots

1) The socalled aggregation observed first by Darwin with the tentacles of
Drosera is a different process, but Bokorny has shown, that there exist certain points
of connection between the two phenomena. Compare Pringsheims Jahrb. Vol. 20
p- 465.

2) We have however observed silver-reduction in black dots with spores of
Gymnosporangium, and with Saccharomyces cultivated at low temperature in a
solution free of sugar.

AS RESERVE-MATERIAL IN PLANTS. 35

of Poa; shoots of Pisum and Vicia; leaves, stems, and flowers
of Tussilago, Ranunculus, Veronica, Convallaria.

The active albumen may serve not only for the growth of
protoplasm, but also for the formation of enzymes. It may be
changed into passive albumen and then form the aleuron grains
and protein crystals, as seems to be indicated by an observation
of Peters, who reports: ‘‘ The formation of protein crystals
proceeds in cells of Sparganium and Carex in, the interior of a
dvop-like accumulation of protein matter by the crystallisation-
process.”") The passive proteids as reserve-material in the
seeds are most probably always products of change of previously
formed active proteids.—

In the animal kingdom no such highly labil, dissolved
vesevve-proteids, are found; at least the coffein-reaction cannot
be obtained. Easily changeable albuminous substances of
another character as the active albumen described occur; the
proteids of the bloodserum causing the artificial and natural im-
munity, as the highly important investigations of Emmerich,
Tsuboi, Buchner have shown, and the poisonous proteids of snakes
and of the blood of eels may be mentioned in this regard. Also
the enzymes or soluble ferments belong to the labil proteids.
I have observed, that these latter substances lose their
efficacy by treatment with a neutral diluted solution of formic
aldehyde, which may be considered as an indication that labil
amidogroups play an important role in their active condition.” —

The mode of action of coffein or antipyrin upon the vegetable
objects mentioned above, consists probably in the formation of
very loose combinations of these bases with the active albumen,
whereby the original chemical nature of the latter is otherwise
not altered, combinations which are less soluble than each of the
constituents for itself. But also the hypothesis is admissible,
that these bases lead mainly to a loose kind of polymerisation
by an irritating influence. Be this as it may, at least it cannot
be disputed that the original condition is restored by washing out

1) Botan. Centr. Vol. 48, p. 181. Interesting are also some observations of
Molisch, Mikosch and Chmielewsky, relating to the occurrence of spindle-and ringlike
bodies of albuminous matter in the leaves of Epiphyllum and Oncidium, Bot. Centr.
1890, II. 341.

2) O. Loew, Jahresbericht f. Thierchemie, 1888, p. 273.

32 ACTIVE ALBUMEN

these bases, which remarkably easily and without producing
injury can penetrate the living protoplasm. It is different how-
ever, if we apply stronger bases (and their salts), as for instance:
guanidin, methylamin, propylamin, anilin, toluylendiamin, atro-
pin, amarin, piperidin, coniin, nicotin, chinin, strychnin, morphin,
codein, chinolin or pyridin in 0,1-0,3 percent solutions. Here
also we observe granulations, but they do not coalesce to droplets,
and solidify soon, becoming entirely insoluble in water. The
protoplasm itself is also attacked with more or less energy by all
these bases and killed quickly by chinin and strychnin, more
slowly by morphin, atropin or pyridin.‘,—Of inorganic bases:
ammonia and its carbonate, hydroxylamin, caustic soda, and
caustic potash produce like results: very minute granulations
and quick death of the cells. But these bases have to be applied
only in 0,1-0,or per cent solutions for a production of a more
intense granulation; a I per cent solution of ammonia will yield a
lesser granulation, than a 0,2 p. mille solution; and a 5 per cent
solution will change the active albumen so rapidly that no granu-
lations at all result. Very dilute caustic potassa produces a
more intense granulation than a corresponding solution of caustic
soda; but neither carbonates nor secondary phosphates of these
two bases, nor limewater can produce any granulations worth
mentioning.

We have seen, that a highly labil, easily changeable albumi-
nous substance is often stored up in plant-cells, that it is used
up in growth and multiplication of cells, and that it behaves
towards ammonia and alkaline silversolution like an aldehyde.
We have found on the other hand (Chapt. IIJ), that a series of
toxicological facts point to the presence of aldchyde-groups in the
proteids of the living protoplasm. This leads us to the logical
conclusion, that our labil reserve-albumen is the very substance
which yields by the organisation-process the living protoplasm,
that it is active albumen, chemically but not morphologically
identical with the active albumen of the living protoplasm. That
this active albumen in the not-organised state is a little less
labil, than in the organised state is intelligible, it brings on

1) Nitrogenous substances deficient of basic qualities do not produce proteo-
somes, as: Skatol, hydrobenzomid, dimethyloxypyrimidin, leucin, tyrosin, asparagin,
allantoin, kreatin.

AS RESERFE-MATERIAL IN PLANTS. 33

some reactions, of which the living protoplasm is not capable on
account of changing too quickly.—As to the great difference
between the active and the passive albumen, convincing evidence
has been furnished, and we recall here the more striking points
of this chemical difference :

1. The power of combining with water is greater with the
active than with the passive albumen;

2. Coffein and antipyrin exert action upon the active, not
upon the passive albumen ;

3. Alcohol of 10-20 per cent, vapors of ether, very dilute
acetic acid, change the active, but not the passive albumen;

4. Active albumen absorbs ammonia and turns thereby in-
soluble, while passive albumen remains indifferent.

5. Active albumen reduces highly diluted alkaline silver
solutions, passive albumen does not.

On the Poisonous Action of Di-cyanogen.

BY

O. Loew, and M. Tsukamoto.

Modern investigations lead to the conclusion, that the proto-
plasm of living cells is a labil organisation of labil material.”
According to the theory advanced by one of us (L.) the great
lability of the albuminous matter composing the living protoplasm
is caused by the simultaneous presence of aldehyde- and amido-
groups. If this is correct then every substance that can react in
a high dilution upon aldehyde- or amido-groups must be poison-
ous for every living cell and organism, from the lowest fungus to
the highest animal. Many toxicological facts and recent ex-
periments are in accordance with this conclusion. The highly
poisonous action of hydrocyanic acid may be best explained by
its easy reaction with aldehyde-groups, while the poisonous
action of di-cyanogen can be best explained by its action on
labil amido-groups. The former reaction is represented by the
following equation:

=0 ae
(x) CL +HCN = (a)-C=ON ,
and the cyanogen reaction by the following example:
CN
2(CoHjNH,)+ | = SuNDC—CInmy

However, the poisonous action of these two substances has been
studied so far only in regard to vertebrate animals, and in regard
to the lowest forms of life only the action of hydrocyanic acid on
yeast cells has been the object of closer observation. It seemed,
therefore, of some interest to compare representatives of all
kinds of living organisms as regards their behaviour to the two
above stated substances.

We prepared solutions of di-cyanogen and of hydrocyanic
acid of a certain strength. The di-cyanogen was prepared in

1) Compare O. Loew, Natiirl. System der Giftwirkungen, Munich, 1893.

ON THE POISONOUS ACTION OF DI-CYANOGEN. 35

the usual way by heating cyanide of mercury; the aqueous
solution, obtained by passing the washed gas in water, was
analyzed in the following way:—z2o c.c. of that solution were
mixed with 2—5 c.c. of concentrated soda lye, and when, after
some time, the cyanogen smell had disappeared, a silver-nitrate
solution was added and afterwards supersaturated with nitric
acid. The washed cyanide of silver was weighed on a dried
filter. The calculated quantity of cyanogen was multiplied by
2, because half of the di-cyanogen having been converted into
sodium cyanate was excluded in this determination. Thus
was found in one case the percentage of cyanogen 0.37%, in
another preparation 0.64% and in the third 0.39%. These
solutions were kept cool, generally below 10°C, and were used
only during the following four or five days for experiments,”
because the solutions of di-cyanogen undergo a gradual decom-
position. In comparing the di-cyanogen solutions with those
of hydrocyanic acid, solutions of equal strength have been used,
starting with the fact that two molecules of hydrocyanic acid
(54) act upon two aldehyde-groups, while one molecule of di-
cyanogen (52) reacts upon two amido-groups.

ACTION OF DI-CYANOGEN UPON MICROBES.

To 50c.c. of the cyanogen-solution of the dilution of 1: 5000
a drop of putrid meat-water was added, and after standing 24
hours, a sterilized peptone-solution was infected from this aqueous
liquid. After standing 8 days there was no bacterial development
and no putrid smell perceptible. In the control experiment, in
which water was used instead the cyanogen solution, after 3 days
a strong development of bacteria with a putrid smell was observed.

In the second experiment in which was compared di-cyanogen
and hydrocyanic acid both in dilution of 1: 10000 a bacterial
developement was observed in both cases, but not till 4 days
later than in the control case, which seems to indicate that the
bacilli themselves were killed but not the spores. In the third
experiment some peas were placed in a cyanogen solution and in
a hydrocyanic acid solution of equal strength, both of them in
a dilution of r:5000. After three days a very strong develop-

1) The Nessler’s test showed us, that during the time, we experimented with
these solutions, no decomposition with formation of NH; could be observed.

36 ON THE POISONOUS ACTION OF DI-CYANOGEN.

ment of bacterial vegetation was seen in the control flask, while
in the flask of hydrocyanic acid solution a slight incipient
turbidity indicated a beginning of vegetation, and in the case
of cyanogen the liquid remained entirely clear and unchanged.
Several other experiments have been made in a dilution of 1: 5000
and 1: 3000, and in each case the poisonous action of the two
substances upon bacteria was clearly observed.

In another experiment, 1 gram of peptone was dissolved at
ordinary temperature in 100 c.c. of a 1: 1000 solution of hydro-
cyanic acid. After filtration, it was infected from putrid meat.
At the same time, 1% solution of peptone was infected from the
same source. After standing five days the latter solution showed
a putrid smell, and white flocculent masses consisting of numer-
ous bacteria, while in the first case no trace of turbidity or of
putrid smell could be observed and no bacteria were found on
examining under a microscope.

In a further experiment, 1: 1000 solutions of di-cyanogen and
hydrocyanic acid containing 1% of kreatin, 0.1% of di-potassium
phosphate and a drop of a 10% solution of magnesium sulphate
were infected from a vegetable matter in lactic fermentation.
After standing a fortnight in closed vessels at ordinary temper-
ature, these liquids") were perfectly clear and free from bac-
teria,”) while the control solution showed a strong development.

The poisonous action of hydrocyanic acid for bacteria was
also proved by the observation of Liebig,*») that blood does not
undergo putrefaction for a considerable time, when 1/1000 of its
weight hydrocyanic acid is added. He also observed, that yeast-
water did not putrefy when some hydrocyanic acid was added
to it.

ACTION OF DI-CYANOGEN AND HyprocyANIc ACID
UPON YEAST CELLS.

Some thick yeast’) obtained fresh from a brewery was
mixed with a di-cyanogen solution of 1: 2500 and after frequent

1) At that time the di-cyanogen and prussic acid remained unchanged and were
perceptible by smell.

2) According to E Schaer the development of mould fungi can be prevented by
hydrocyanic acid in a dilution of 1: 10000 (Zeit. f. Biologie 1870, Bd. 6, S. 509.)

3) Ann. der Chemie u. Pharmacie 1870, Bd. 153, S. 137 etc.

4) The yeast-mass of the size ofa pea corresponding to in average 0.07—o.1 grams
dry matter was shaken with 50 c.c. of the solution.

ON THE POISONOUS ACTION OF DI-CYANOGEN. 37

shaking the supernatant liquid was after 24 hours poured off.
The sediment of yeast was then shaken with 5 c.c. sterilized
Pasteur’s solution’) and placed in a narrow test-tube. No trace
of fermentation was observed even at a temperature of 30°C,
while in the control experiment a vivid fermentation was
going on after half an hour. In a second experiment about the
same amount of yeast was mixed with a di-cyanogen solution in
a dilution of 1: 5000, respectively 1: 10000 (30 c.c.), and left to
stand, being frequently shaken, for 20 hours. The liquids were
poured off from the yeast and the latter was mixed with Ioc.c.
of sterilized Pasteur’s solution at 25°C. No fermentation was
observed after 3 hours with yeast that had been treated with
di-cyanogen solution of 1: 5000, but later a few small gas-bubbles
developed, although even after one day the development of
gas was very slight, while in the control experiment after 15
minutes a strong fermentation had set in. On the other hand,
in the case of the yeast having been in contact with the cyanogen
solution of 1: 10000 only 20 gas-bubbles developed in 1 minute
after being 15 minutes in Pasteur’s solution at 25°C, but more
than 200 gas-bubbles were observed in the control case in the
same duration of time.

In another experiment it was found that every trace of the
fermentative action was annihilated after 24 hour’s contact of
yeast with di-cyanogen solution of 1: 2000, while in several ex-
periments with hydrocyanic acid the fermentative action was not
annihilated. When the yeast was suspended for 24 hours ina
sugar solution”? containing hydrocyanic acid in a dilution of
I: £000, as long as hydrocyanic acid was present no fermentation
was observed, but as soon as it was removed and the yeast settled
at the bottom was shaken with plain sugar solution, the fermen-
tation was almost as energetic as in the control mixture. This
observation confirms those of Schénbein,*) Schaer*) and Liebig
(I. c.), that hydrocyanic acid of a certain strength will paralyze

1) Instead of the tartrate of ammonia however extract of meat was used.

2) This solution contained 5°/, cane sugar, 1°/, extract of meat, 0.1°/, KH2,PO4
and a trace of magnesium sulphate and was mixed with so much prussic acid that
the percentage corresponded exactly with the stated amount.

3) Zeitsch. fir Biologie 1867, Bd. III, S. 142.

4) Ibid. 1870, Bd. VI, S. 503.

38 ON THE POISONOUS ACTION OF DI-CYANOGEN.

the fermentative action of yeast without killing it. But we have
found further that if the hydrocyanic acid of the same strength
remains in contact with yeast for more than one day, or if, in-
stead of hydrocyanic acid solution of 1: 1000 a stronger solution
is applied, as 1: 400, it will damage the fermentative activity so
much that afterwards the yeast produces only a very faint fer-
mentation. It seems to us that only a few yeast cells escaped
death under these circumstances. We have found furthermore
that 2 day’s contact with hydrocyanic acid in a dilution of r:
5000 with frequent shaking, will kill almost all the cells, so
that after pouring off the hydrocyanic acid, the yeast being
shaken up in sterilized Pasteur’s solution showed only very
slight indications of fermentative action.

If we compare this result with our further observation that
the fermentative action is not suppressed when we add to the mix-
ture of yeast and Pasteur’s solution prussic acid in the proportion
of 1: 5000, we are led to the conclusion that in all probability
the hydrocyanic acid upon entering the yeast cell undergoes
decomposition before it can act poisonously. If, however, the
yeast is not in fermenting activity, the entering hydrocyanic acid
is not so quickly decomposed, and therefore acts as a slow poison.
It may perhaps be assumed that the change consists in the addi-
tion of H,O, forming thereby formamid :

=O
HCN +H;0 = HC=0,,..

ACTION OF DI-CYANOGEN AND PRussic ACID
UPON ALGAE.

If threads of Spirogyra communis are treated with a solution
of di-cyanogen of 0.39%, instantaneous death with contraction and
turbidity of the cytoplasm is observed. In solutions of di-cyano-
gen, respectively hydrocyanic acid of 1: 1000, no noxious effect
was seen after 30 minutes, but after 4 hours most of the cells in
the cyanogen solution were killed; in the prussic acid solution,
however, a much smaller proportion of cells. After 15 hours, all
the cells were dead in both cases. Also solutions of 1: 10000
exerted a poisonous influence after several days", but it could

1) In another case the noxious effects of the two substances in the same dilution
were observed after 20 hours; on treating with very dilute methyl violet the cells
of the algae in the cyanogen were all coloured, but of those in the prussic acid some
cells not, showing these were still alive.

ON THE POISONOUS ACTION OF DI-CYANOGEN. 39

be distinctly seen that HCN was less noxious than di-cyanogen.
In a dilution of r:100000 of di-cyanogen Spirogyra, Oscillaria
and Diatoms were seen alive after several days.

ACTION OF DI-CYANOGEN AND HypDROCYANIC
ACID UPON PH#NOGAMS.

In a solution of prussic acid 1: 1000 which had no reaction
upon litmus paper, some young radish plants 4 cm. high were
placed. After 15 hours, leaves and stem had withered, lost
the turgor, and commenced to dry up. Di-cyanogen solution of
I: 5000 acted almost as quickly upon young barley plants with
stems I0 cm. high. Even ina solution of di-cyanogen of 1: 10000
young barley plants 5—6 cm. high were attacked after 3 days,
turned yellowish, and commenced to dry up. In 200 c.c. of cy-
anogen solution 1: 25000, to which were added the necessary
mineral salts in the usual proportion, was placed a young water-
cultured lupin-plant with a stem 8cm long. After 20 hours the
stem was bent down, and also the leaves without any turgor.
After further 15 hours, however, the plant seemed to recover, but
only for a short time, for 20 hours later the plant commenced to
dry up. In another experiment, a stem of Narcissus (N. tazetta,
var. chinensis, Roem.) was cut immediately above the bulb and
placed in a solution of di-cyanogen in r: 10000. In this case no
noxious effect was observed, evidently there was so much dis-
solved albuminous matter encountered in the juice by the cyano-
gen that this poison could not reach enough cells to kill the plant.

Some experiments were made with seeds that were just
beginning to germinate after having been soaked in water. Thus
the seeds of peas, turnips, radishes, and barley were treated
with cyanogen-respectively hydrocyanic acid solution of 1: 5000,
in each case 12 seeds being placed in 5o0c.c., and left to stand
for 3 days in closed vessels. Control experiments with distilled
water were also made. After pouring off the liquid, the vessels
were left to stand at 15°—20°C; after 24 hours great differences
were noticed: in the control experiments the seeds developed in
the normal way, the others, however, remained stationary’).

1) Schoenbein treated some seeds for half an hour with very diluted prussic
acid and found that most of these seeds did not germinate any more. Some of the
seeds, however ,germinated imperfectly (Zeitschrift. f. Biol. 1867, Bd. 3, S. 1423).

40 ON THE POISONOUS ACTION OF DI-CYANOGEN.

Only in one case, where the turnip seeds had remained in the
hydrocyanic acid solution, the development of the cotyledons
was observed after 6 days while the life of the root-germ was
entirely destroyed. Here probably the hydrocyanic acid did not
enter far enough into the seeds to kill all the cells, while the
root-germs were more exposed.
ACTION OF DI-CYANOGEN AND PRussic AcID
UPON LOWER AQUATIC ANIMALS.

When under the microscope infusoria, nematodes, rotatoria,
annelides, Copepodes, and Ostracodes were brought into contact
with several drops of dicyanogen-solution in a dilution of
1: 2000, all the life was annihilated within two minutes, while
in a parallel experiment with an equal dilution of CNH the
animals were alive after 30 minutes, but some time afterwards
died under convulsions’). In di-cyanogen solution of 1: 10000
(200 c. c.) the death of these animals very soon took place, also;
only the worms lived longer, but after 15 hours they also died.
In hydrocyanic acid of the same dilution the animals lived
longer than in the cyanogen solution, and still after 15 hours
ostracodes showed convulsions in their legs. Even in the dilu-
tion of r:100000 of dicyanogen all infusoria with the excep-
tion of some monadines were killed. Also the copepodes died,
but the ostracodes and worms still showed convulsions. In
hydrocyanic acid of the same dilution many infusoria were
alive after 5 hours, and some even after 18 hours. Ina second
experiment with some mud rich in infusoria (Pavamaecitum)
some individuals were found alive after one hour in cyanogen
solution of 1: 20000, but all were killed after 20 hours even in
the solution of 1: 100000, while in the case of hydrocyanic acid
in the same dilution many infusoria were left alive.

We see, therefore, that all these experiments prove the di-cy-
anogen to be a stronger poison than hydrocyanic acid; and,
therefore, the results of B. Bunge*) with vertebrate animals, that

1) The assertion of Schaer (Zeit. f. Biol. 1870, Bd 6, S. 511), that infusoria are
not killed by HCN is certainly erroneous. It may be that only an experiment of
short duration under the microscope had been made. In our experiments with
infusoria ordinary well-water was used for the dilutions.

2) According to B. Bunge (Arch f. exper. Path., Bd. 12, S. 41-75, or, Jahresb.
f. Anat. & Physiol., 1879, Bd. 8., Ab. 2, S, 187), small doses of cyanogen produce
general central paralysis. With warm blooded animals dyspnoe, and paralysis of
respiration are produced. For killing a cat 0.02 grm. cyanogen are necessary while
of prussic acid 0.004 grm. suffice.

ON THE POISONOUS ACTION OF DI-CYANOGEN. 41

hydrocyanic acid acts about 5 times stronger than di-cyanogen,
seems to us exceptional. One experiment, which we made
with two white rats, proved indeed that here hydrocyanic acid is
much stronger. The one rat weighing 124 grams received by sub-
cutaneous injection I c.c. of hydrocyanic acid solution I: 10000,
the other rat weighing 156 grams received an equally diluted
solution of dicyanogen (1 c.c.). The former rat was found
dead after ro hours, the latter, however, remained alive and
healthy. The fact that the di-cyanogen here acts less powerfully
is probably due to the fact, that free cyanogen can directly act
upon proteids in solution,’) and therefore can be changed to a
great extent in the blood and lymph, before it reaches the
nervous centres. Hydrocyanic acid does not act at all upon the
ordinary passive proteids, and can, therefore, without being pre-
vented by the serum albumen, reach the nervous centres,and act
there upon the active labil proteids of the living protoplasm.

As the general result we see, that the prediction that free
cyanogen would prove a general poison for all kinds of living
organisms, has been confirmed. At the same time, it has been
shown that also hydrocyanic acid is a general poison and we
hope therefore that the explanation, which we have given
above, will prove more durable than the former unsatisfactory

hypotheses.”

1) Compare: O. Loew, on the action of di-cyanogen upon albumen, Journ. f.
prakt. Chem., 1877.

2) The former hypothesis that hydrocyanic acid acts poisonously merely on ac-
count of combining with the haemoglobin has been proved erroneous (J. Belky,
Jahresb. f. Thierchemie, 1885, Bd. 15, S. 155).

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The Energy of the Living Protoplasm.
BY
Dr. Oscar Loew,

Professor of Agricultural Chenustry.

CHAPTER V.

THE FORMATION OF PROTEIDS IN PLANT-CELLS.

The formation of albuminous matter in plant-cells is cer-
tainly one of the most important processes in the domain of
general physiology, as it is upon this process that depends the
growth of protoplasm, the multiplication of cells, the develop-
ment of plants; upon plant-life again subsists all animal life.
This remarkable synthesis takes place in fungi just as well as in
chlorophyllaceous plants; but while the latter use principally
the carbohydrates as sources of carbon, the former make use
of a variety of organic compounds. We consider first :

THE FORMATION OF PROTEIN IN MICROBES AND MOULD-FUNGI.

Among all the fungi the microbes are especially remarkable
for the intensity of their chemical activity. Oxidations and
decompositions take place on an extensive scale. Numerous
organic combinations are easily split and, under atomic migra-
tions, substances of a more solid structure are formed: the
products of fermentative actions. And amid this destructive
activity there is built up in the interior of the cells the most labil
of all combinations, the active albumin, which is organised into
living protoplasm. And this is done under favorable conditions
with such rapidity that one cell yields by multiplication in 24
hours more than one trillion of new cells. What an energetic
synthesis of protein-matter, of preparation of living protoplasm,
of living cells! The destructive operations are necessary to
carry on the synthetical work, furnishing not only energy but
also the required atomic groups; and consisting either in oxida-

et Me

44 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

tions or in fermentations. We leave in this chapter, however,
the fermentative activity out of consideration, and mainly treat
of the protein formation and nourishment of the aévobic microbes
and mould-fungi:—We observe here that a great number of
organic compounds of different chemical constitution may serve
for nourishment and development. There can be hardly any
doubt but that in all these different cases the same proteids
result, otherwise the structure and the functions of the pro-
toplasm formed would show variations, new species would spring
into existence with ease, according to difference in food. We
find however the same species of a bacterium’) or of a mould-
fungus, whether we nourish them once with peptone, or with
sodium tartrate as a source of carbon, whether we offer glycerin,
glucose or lzvulose, xylose or arabinose, glycol or chinic acid.
It makes no difference in the resulting species, whether we offer
once nitrates, the next time leucin or a third time betain as
a source of nitrogen.

This circumstance teaches us not only that the proteids
and protoplasms formed from different food, remain in one
species the same, but also that the formation of protetds must
commence with relatively simple atomic groups, that ave prepared from
the most different kinds of substances by oxidation and decompost-
tion.”

Certain combinations are excellent nutrients, like peptones,
others poor ones, like valerianic acid, others again do not serve
at all as sources of carbon, as oxalates or pyridin salts, and others
are poisons, as phenylhydrazin. Small chemical changes may
convert a nutritive compound into a poison and the poison again
into an indifferent body. These qualities depend upon the chemi-
cal constitution and are to some extent merely relative concep-
tions determined by the degree of concentration. Glucose

1) The characters of bacteria may however be modified by changing the condi-
tions of cultivation; but whether such modifications would lead in course of time to
new species, which would be of high importance in connection with the problem of
evolution, remains to be investigated.

2) Pasteur was the first, who recognised the capability of mould-fungi and bacteria
to grow in solutions free from protein compounds, i. e. to form protein and protoplasm
from simply constituted combinations, as tartaric acid, ammonia and sulfates. Up
to that time (1858) the opinion prevailed that fungi, like animals, could only subsist
upon proteids.

THE FORMATION OF PROTEIDS IN PLANS-CELLS. 45

can be utilised in higher concentration than sodium acetate, and
this in a higher one than acetic ester or phenol, the latter form-
ing in 0,08 % solution a meagre food for a micrococcus,” although
in higher concentrations a poison for all kinds of bacteria. The
more easily a compound is attacked by the cells, the quicker the
development of new cells.

As sources of carbon: alcohols, acids, ketons, aldehydes,
carbohydrates, esters and bases can serve. As sources of nityogen :
nitrates, ammonium salts, amido-acids, ureas, guanidins, nitriles,
amines, ammonium bases can serve. As sources of sulfur may be
utilised not only sulfates but also organic sulfur compounds, as
sulfons, sulfonic acids and probably also sulfides (methyl sulfide).
Our observations in regard to the sources of carbon teach us :—

1) The nutritive quality of acids is enhanced by the en-
trance of alcoholic hydroxyl-groups; thus lactic acid is superior
to propionic acid.

2) The nutritive value of alcohols is increased with the
number of the hydroxyl-groups: glycerin is better than propyl
alcohol.

3) The presence of aldehyde-or keton-groups increases the
nutritive qualities: glucose is better than mannite; acetyl acetic
ester is better than acetic ester.”

4) The lower alcohols may be used in higher concentration
than the higher ones (amyl alcohol).

5) The lower members of the fatty series are more easily
assimilated than the higher members; sodium acetate being far
superior to sodium valerianate.

6) The unsaturated ring systems are generally not favora-
ble; for instance antipyrin and dimethyloxypyrimidin appear
to be entirely unsuitable and while benzoic and salicylic acid
are very poor sources of carbon, phenyl acetic acid, containing a
CH, group, is far better, and chinic acid, containing a saturated
benzol-ring with 4 CHOH groups is a very good source.

To carry out such experiments, mineral solutions are
prepared containing 0,1—0,5% dipotassium phosphate, o,o1—
0,05% magnesium sulfate and 0,1—o0,2% potassium nitrate or

1) Compare Nédgeli, Ber. Bayr. Akad. d. Wiss. 1879.
2) I have here operated with 0,1 °/9 solutions, Biol. C, 10, 585.

46 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

di-ammonium phosphate.) After addition of the organic
nutrient to be tested, the liquids are inoculated with bacteria.
In those cases, in which the growth of a specific bacterium has
to be tested, a previous sterilisation is of course necessary.
An incipient turbidity, the formation of flocculi or of a thin
film and the microscopical examination will very soon indicate
that the bacteria have grown and multiplied. Organic bases
are best neutralised with phosphoric acid, while acids are best
applied as sodium salts.

It is, however, not only of interest to decide, which combina-
tions can serve for nutriment, but also to elucidate the reasons,
why often closely related compounds behave far differently
from each other, and why certain substances, which are in
neutral solution by no means poisonous, cannot be used as
food. We find for instance that pyridin, pinacon, dimethyloxy-
pytimidin, ethylendiamin, amidoacetal, glyoxal, meconic acid,
oxalic acid in dilutions of 0,5 9% do not support bacterial growth,
and acetoxim, diacetonamin, citraconic and maleic acid permit
only with difficulty after a series of weeks a gradual develop-
ment.” These compounds are therefore not well suited for
the preparation of those atomic groups that serve for the
formation of proteids. Control experiments with addition of
0,2% peptone indicated, by the rapid development of bacteria,
that neither of these substances are poisonous in such a degree
as to kill the bacteria if well nourished.

The fact that fumaric acid supports bacterial life so well,
that a few days after the infection the liquid swarms with
numberless bacteria, while it takes 4 weeks with the isomeric
maleic acid before a slow development, after repeated infection,
takes place is of biological interest. Whether here a gradual
adaptation took place or only specific germs were developed,
remains to be decided. In the case of citraconic acid no
development for six weeks was observed; finally, after repeated
infections a scanty vegetation set in, consisting of apparently
only one kind of bacterium, forming short thick rods often in

1) Calcium salts are not necessary for the life and development of the lower
fungi, Compare O. Loew, On the functions of calcium and magnesium salts in
plants, Flora 1892.

2) O. Loew, Centr. f. Bacteriol. 22, 361

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 47

lively motion. At this time fumaric acid in a control solu-
tion had been entirely decomposed and the bacterial vegetation
formed slimy flocculi at the bottom of the flask.”

Of interest is the contrast between the moderate nutritive
qualities of aceton and the exceedingly poor results of diaceto-
namin, further that between glycol and the non-nutritive
pinacon, i.e. tetramethylglycol, while trimethylcarbinol forms
again a moderately good source:

ey a CH
HC OH (CH)© On CH, >C—OH.

H,C—OH (CH,),C—OH CH;
Glycol. Pinacon. Trimethylcarbinol.

While furthermore betain forms a better source than gly-
cocoll, we find that trimethylamin is not so favorable as methyl-
amin. If we compare the latter solutions after a strong
bacterial turbidity had set in, we have to dilute it at least
5 times with water until the degree of turbidity has reached
the weak turbidity of the trimethylamin solution.”

H3N
CMs CH; CH; tia.
\N CH SN CH; SN CH2—CO | CH.
aa CH. He 4 lin aa | |
po ee SS aeeeee = O—CO
Methylamin. Trimethylamin. :
: Betain. Glycocoll.3)

To the nitrogenous compounds which neither serve as a
source of carbon nor as a source of nitrogen, belongs dimethy]-
oxypyrimidin® in 0,2% solution, while coffezn serves not only as
a source of carbon, but also as a source of nitrogen.

7 N—C—CH; CH3.N—CH
GEC al
> N= ACH CO C—N—CH;
—— 9 __——. ie SCO:
Dimethyloxypyrimidin. CH3.N—C= =N
Coffeine

If we compare the monovalent alcohols, from methyl to

1) In this case as in similar ones the principal bacillus developed resembled
Bacillus liquefaciens fluorescens.

2) Both these bases were applied in 0,5 0/o solutions neutralised with phosphoric
acid, and were infected with Bacillus methylicus. O. Loew, Centralbl. f. Bacteriol.
12, 465.

3) As regards this formulation of glycocoll, compare Yoji Sakurai, Journal of
the College of Science, Vol. 7.

4) The oxypyrimidins first prepared by A. Pinner have a phenol-like character.
Deutsch. Chem. Ges. 18, I owe Prof. Pinney many thanks for providing me with a
number of pyrimidins,

48 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

amyl alcohol, we observe that the higher members of the series
have noxious qualities. We must apply much higher dilutions
to be able to raise a bacterial vegetation. While methyl alcohol
in 1% solution, containing 0,1% K,HPO,, 0,1 % (NH,).HPO,
and 0,o1% MgSO, develops easily bacterial growth, we have to
use amylic alcohol in dilutions of 0,1 % to make this possible.)*

Of considerable interest is the decrease in the nutritive quali-
ties of the fatty acids with the zzcrease of their molecular weight.”
If we prepare for instance 0,5 % solutions of sodium acetate and
of sodium valarianate and add to each of them 0,2% K,H,PO,,
0,2% KNO,'and 0,05 % MgSO, and infect a set of these solutions
with spores of Penicillium, with Saccharomyces Mycoderma and
with bacteria from putrid meat, we observe with acetate of sodium
after three days a considerable development, while with the
valerianate merely a slight turbidity is visible. After another
3 days a most luxuriant development of Penicillium, Saccharomyces
and of large bacteria takes place in the acetate, while neither
Penicillium nor Saccharomyces, but merely moderate bacterial
vegetation consisting of large bacilli forming zooglcea is observed
with the valérianate.

In a like manner it was observed that Penicillium did not
grow in 0,1 % solution of lecithin,*) but only bacteria to a mode-
rate extent; this vegetation made the impression of a pure
culture, although the infection was made from putrid meat con-
taining various kinds of microbes; probably that solution does
not suit for every kind.

The lowest of the fatty acids, formic acid, can, as it seems,
only be utilised by one kind of bacterium,*) evidently a difficulty
being encountered here in transforming it into the suitable
group for synthetical operations. The next related compound,
form-aldehyde, is as such, in the free state, poisonous, but
it ferms combinations with primary sodium sulfite and with
ammonia, which can be utilised as sources of carbon for a
bacillus and fora kind of Dematium.”

1) According to R. Brown (Chem. Soc. Journ. 1886) Bacterium aceti utilises
methyl-, but not amyl alcohol.

2) These observations agree essentially with those of Stutzer, Z. f. physiol.
Chem. 1882.

3) Spores of Penicillium were introduced repeatedly during a period of 4 weeks,

4) O. Loew, Centralbl. f. Bacteriol. 12, Nr 14.

5) Ibid. and Botan. Centr. 1890.

THE FORMATION OF PROTEIDS IN PLANT-CELLS. ze)

I have observed farther that not only methyl alcohol, but
also methylal and methyl sulfuric acid’) may in proper dilution
(0,2-0,3%) be used as food by bacteria, i. e. as material for
building up protein and protoplasm. The group then used for this
purpose must contain only one atom of carbon and cannot be any thing
else but form-aldehyde, the same substance that forms by condensation
various kinds of sugar. Neither acetic, glycolic, nor amidoacetic
acid can be utilised as such, but they may lead by oxidation
to one compound which can be utilised, viz. to form-aldehyde; no
other unsaturated atomic group could result, suitable for
synthesis. This oxidation” in the cells may be expressed by
- the following equation in the case of acetic acid:

CH,.COOH+0,=CH,0+C0O,+H,0

If this conclusion,is correct, then we can understand, why
substances containing the group CHOH are very favorable for
nutriment and why the useful qualities increase with the number
of these groups (the polyvalent alcohols, the polyvalent acids).
We can understand furthermore, why such substances are
capable of nourishing certain bacteria endowed with fermentative
properties, even in the absence of airy, while compounds without this
group can be used as food only in the presence of air, oxidation
being then necessary to produce this CHOH-group or the
isomeric formic aldehyde. But can that conclusion be admitted
if formic aldehyde is a poison? No doubt this seems an objec-
tion of weight, but if we consider how easily the formic aldehyde
is changed under condensating influences and how indifferent
certain compounds of this aldehyde are, the objection no longer
appears so serious ; we must only adopt the view that the formic
aldehyde undergoes rapid transformations and that no molecule
formed remains unchanged for a second.°?

1) In this case an alkaline reaction is necessary for obvious reasons.

2) Oxidation by respiration has here evidently not only the physical role of
yielding energy, but also a direct chemical function in preparing the suitable group
for synthesis.

3) Compare O. Loew, Ber. d. Deutsch. Chem. Ges. 22, 484. Synthetical
processes require substances of a certain lability; very reactive substances however
are more or less poisonous. The objection of some botanists to the view of the
formation of sugar from form-aldehyde in plants is not more tenable since Bokorny
has shown that the combination of form-aldehyde with primary sodium sulfite can
be utilised by plants for the production of starch. Landw. Jahrb. 1892.

50 THE FORMATION. OF PROTEIDS IN PLANT-CELLS.

We have mentioned above that a certain kind of bacillus”)
can utilise formic acid (as sodium salt) as a source of carbon
which is of special interest. Here the first step is most probably
a transformation into glyoxylic acid and a subsequent splitting
of the latter into form-aldehyde and carbon dioxide,*) as may be
expresed by the following equations ;

2 H.COOH = 0:CH.COOH -+ H,0
O:CH.COOM =" CH0 -- ce

The necessary energy is evidently furnished here by the
oxidation of a certain portion of the sodium formiate into
sodium carbonate, clearly indicated by the gradual increase of
the alkaline reaction. The more energy, however, a compound
yields in being utilised for protein formation, the better the
effect must be; we can therefore understand, why methyl alcohol
is a better source of carbon than form-aldehyde (in the shape of
CH,OH.SO,Na) or formic acid. Methyl alcohol yields by its
transformation into form-aldehyde an amount of energy, that form-
aldehyde cannot furnish; the necessary energy must be gained
here by oxidation of such molecules to carbon dioxide and water.

We understand now why oxalic acid, parabanic acid, urea
or guanidin cannot be used as sources of carbon; this is because
these compounds cannot be transformed by bacteria into form-
aldehyde. If pyridin, pinacon or dimethyl-oxypyrimidin will
not serve, it is very probably on account of the considerable
resistance they offer to oxidising influences. If amidoacetal,
glyoxal, or ethylendiamin are incapable of yielding a bacte-
rial vegetation, and diacetonamin and acetoxim apparently

I

do so only with great difficulties, we might suspect a certain
degree of poisonous character, that cannot make its appearance
in presence of a nutrient like peptone.*) But for the difference
of physiological value between the stereo-isomeric maleic and
fumaric acid, we have at present no explanation.

As a general rule we find that compounds, containing the

1) This bacillus, which I named Bacillus methylicus, can also utilise the
combination of form-aldehyde with primary sodium sulfite, CHz0H.SO3Na oxyme-
thylsulfonate of sodium in 0,5°/, solution.

2) Compare Koenigs, Ber. Deutsch. Chem. Ges. 25, 801.

3) Some of the combinations of this group might perhaps be utilised in much
higher dilutions than tested (0,5°/.)-

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 51

groups CH,, CH,, CHOH, CH.OH canbe used as sources of
carbon, if they neither act poisonously nor offer too much
resistance to the attacks of bacteria. If the atomic group
CH: CH is utilised, as in the case of fumaric acid, we might
easily explain this by the conversion into CH..CH, OH, a
molecule of water being taken up. In the following table
are enumerated a number of compounds that form very good
sources(I), moderatly good sources(II), very poor sources (III)
and such, as far as observations reach, cannot be utilised as
sources of carbon(IV).

I II III IV
Glycerin Methyl alcohol Phenol Pinacon
Mannite Aethylenglycol Acetoxim Sulfonal
Sugars Aceton Diacetonamin Amidoacetal
Lactic acid Acetic acid Valerianic acid Oxalic acid
Succinic ,, BiimMaTic sss Maleic Pe Meconic ,,
Tartaric ,, Pyruvic 3 Citraconic ,, Picric on
Citric a Laevulinic ,, Benzoic: ,; Antipyrin
Betain Glycocoll Lecithin Dimethyl-oxypy-

rimidin

Alanin Methylamin Trimethylamin Aethylendiamin
Leucin : Cholin Strychnin Pyridin
Asparagin Allantoin Hexamethylen- Urea

amin
Glutamin Coffein Amidobenzoic Parabanic acid

acid
Kreatin!) Methyl cyanide Glyoxylic acid Guanidin

A most remarkable phenomenon in regard to bacterial deve-
lopment (therefore also to the formation of protein) was first
observed by Ff. Hiippe,”) and later confirmed by Munro and by
Winogradzky, that the nitrifying bacteria of the soil may develop
in solutions destitute of organic matter, and may utilise ammo-
nium carbonate. Hiippe assumed here a decomposition of water,

1) Th. Bokorny and myself have made a series of experiments to nourish also
alyae with organic matters, and observed a much more favorable result with kreatin
and hydantoin than with leucin or urethan. Recently Th. Bokorny extended these
investigations to a larger number of combinations, showing that algae may well
utilise organic matters, although they do not require them for supporting their life.
(Chemikerzeitg, Jan. 1894).

2) Huppe, Biol, Centralbl. 7, 702.

52 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

whose oxygen would produce the nitrification in acting upon the
ammonia, while the hydrogen would act reducing upon the
carbonic acid of this salt, producing thus form-aldehyde, or
carbohydrates. The process, however, would appear simpler if
a part of the hydrogen of the ammonia could be employed for
this reduction. I tried to explain this process by the following
equations’ :

2NH,+20,=2NO.H+4H,
CO.+4H = CH,0+H8H,0,
6CH.O = Cen

This production of organic matter from inorganic without
the aid of the chlorophyll apparatus is certainly most extraordi-
nary.

Some additional remarks may be made on the nutrition of
mould-fungi (Penicillium, Aspergillus, Mucor). Substances sup-
porting the life of aérobic bacteria generally also serve as food for
mould-fungi ; however there exist exceptions: methylamin, methyl
alcohol or sodium valerianate are better utilised by bacteria,
glyoxal better by mould-fungi. Neutral reaction is best suited
for most kinds of fungi, in alkaline liquids however bacteria thrive
better than mould-fungi, while in an acid one the contrary is
observed (with certain exceptions”). In certain solutions the
development of bacteria will prevent the development of mould-
fungi, in other solutions again both kinds of fungi may thrive at
the same time. I infected for instance a solution containing
potassium sodium tartrate 1%, di-potassium phosphate and diam-
monium sulfate 0,1% of each, and magnesium sulfate 0,05 %,
with bacteria of putrid meat and with spores of Penicillium glau-
cunt, Simultaneously, but only the bacteria developed although I
rendered after 2 weeks the solution slightly acid with mono-
potassium phosphate and infected once more with Pemiczlliwm-
spores. On the other hand when glucose was employed instead

1) O. Loew, Ibid. 10, 758 and Centralbl. f. Bacteriol. 9, Nr.20. Warington
expressed recently the same idea (Chem. News 68. 175) and gave the following
equation : :

(NH,4)2CO3+O0=NHy. NOz+CH20+4H20.

2) While Bacterium aceti thrives well in a solution containing several per cent
acetic acid, cases may also be observed, where in an alkaline liquid bacteria cannot
thrive but only mould-fungi.

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 53

of the tartrate, both kinds of fungi developed well simultaneously.

It is of interest to compare the amount of mould-fungus
with the quantity of the compounds used for development; con-
siderable differences are here observed. While isobutylic alcohol
yields only g—10% fungoid matter, asparagin yields nearly 22%;
tartaric acid yields less than succinic, tannin less than sugar.
Albumin in 1% solution will yield about 23 % of its weight, while
a mixture of albumin (1%,) and cane-sugar (2%) nearly 33 %.

To those compounds which cannot be utilised by mould-
fungi belong maleic acid,” citraconic, mesaconic, dibenzylma-
lonic and diethylsuccinic acid. Benzyl-succinic, disubstituted
glutaric acid and oxyisobutylic acid are very poor sources, while
malonic, succinic, monomethylsuccinic and monoethylsuccinic
acids are well utilised.”

But it is not only in regard to the sources of carbon that a
great variety exists; this holds good also in regard to the sources
of mtrogen. Of the great number of the latter compounds we
mention as examples: nitrates, ammonium salts, glycocoll, aspa-
ragin, kreatin, allantoin, methylamin, acetamid, methylcy-
anide, betain, strychnin. Nitrites are in a certain concentration
less favorable than nitrates and are in acid solutions poisonous.
Ferrocyanide of potassium is but a poor source of nitrogen while
hydroxylamin and diamid cannot be utilised at all, being strong
poisons, and azoimid only in high dilutions.*) For the same
reasons, as explained above for the sources of carbon, the nitro-
gen compounds used must be converted first into the one
and the same atomic group, before the synthetical work can
begin; this group is evidently ammonia which, in form of salts,
is not only very favorable for mould-fungi and bacteria, but
also the simplest nitrogen compound that can directly be
utilised.) If organic nitrogen compounds are used as sources

1) E. Buchner B. d. Deutsch. Chem. Ges. 1892 p. 1163.

2) B. Meyer, Ibid, 1891 p. 1071.

3) O. Loew, Biol. Centralbl. 10, 588 and Ber. Deutsch. Chem. Ges. Vol. 24,
p- 2947. Certain fungi, as Saccharomyces Mycodcyma prefer ammonia as source of
nitrogen to amido-acids and peptone (Beyerinck). The common beer-yeast however
can at Jow temperature make better use of the latter than of the former, and cannot
utilise nitrates (4. Mayer). Laurent has shown that these are converted here into
the poisonous nitrites.

4) Nitrates have to be reduced first.

54 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

of nitrogen, the latter has to be split off in shape of ammonia,
before the protein formation can begin. This can be accom-
plished in many cases by simple splitting (acetamid, kreatin,
urea, etc.), in other cases by oxidation, as with leucin, methyl-
amin, betain. Chinin and strychnin are poor sources of
nitrogen, being attacked by fungi with difficulty, and antipyprin
and dimethyl-oxypyrimidin offer so much resistance, that their
nitrogen cannot at all be utilised.

The anaérobic microbes may by reducing influences transform
nitrogen of certain compounds into ammonia, while the aérobic
employ oxidation :

1) H.N.CH;.COOH +2H=NH,+CH,.COOH,
2) H.N.CH:.COOH+30=NH,+2CO,+H,0.

A very remarkable case is the assimilation of free nitrogen
by certain bacteria of the soil as was asserted years ago by
Berthelot? and recently confirmed by Wunogradzki. The free
nitrogen is here probably first converted into ammonium
nitrite,” and the nitrous acid then also rapidly reduced to
ammonia.

The proteids of mould-fungi and of yeasts) contain, like the
other proteids, sulfur which can partially be split off by diluted
alkaline ley in form of sulfide; the sulfur must therefore be in a
very loose form of combination, very probably present as—SH
in the proteins. Sulfates have to be reduced therefore to sulfuret-
ted hydrogen, before the sulfur can be assimilated, and organic
sulfur compounds have to be split before reduction and assimi-
lation take place. My experiments with sulfonal (CH,).: C:
SO,(C,H,). have shown that this substance serves well as a
source of sulfur for mould-fungi and yeasts, in the presence of a
good source of carbon; but in the absence of such a source, sul-
fonal cannot be used, although it contains the methyl and ethyl
group, 1.e. otherwise good sources of carbon; the fungi refuse to

1) Compt. rend. ror.—Also Gautier and Drouin, Ibid, 106.

2) About this transformation with the aid of platinum black, see O. Loew, Ber.
Deutsch. Chem. Ges. 23, 1444.

3) Nencki found in 2 cases no sulfur in the proteids of bacteria,

Mitscherlich found in yeast 0,6°/o sulfur. If yeast be treated with a diluted
solution of caustic potash, a considerable amount of potassium sulfide is formed.

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 55

grow in solutions containing as sole organic matter sulfonal
(0,2 %).

Of course sulfuretted hydrogen is only produced in the neces-
sary amount for immediate need, as accumulation would be
noxious ; experiments intented to prove that H.S as such can be
assimilated, meet for obvious reasons with some difficulties.

Our considerations, therefore, lead logically to the conclu-
sion, that the atomic groups, serving for the formation of proteins
are three very reactive combinations :

Form-aldehyde, ainmonia and sulfuretted hydrogen.

IJ]. THE FORMATION OF PROTEIDS IN
CHLOROPHYLL-BEARING PLANTS.

As chlorophyll-bearing plants produce by assimilation car-
bohydrates, it is natural that such well suited compounds
should form here also the main source of carbon for the synthe-
sis of proteids.’) Nitrates or ammonium salts furnish the nitro-
gen, sulfates the sulfur. The chemical behaviour of the protein
compounds indicates very clearly, that neither the nitrogen
nor the sulfur is connected with oxygen, but only with carbon
and hydrogen. It follows, therefore,- that veduction of the
nitrates and sulfates has to take place, here as well as in the
case with the lower fungi. If all conditions are otherwise
favorable, then the synthetical work proceeds so rapidly that
the intermediate steps cannot be directly traced. From numer-
ous observations, however, the conclusion appears justified
that it is aspavagin to which an important réle must be attributed
in this connection, a conclusion which at first was arrived by
the ingenious Th. Hartig. Borodin, Pfeffer and Kellner declared
asparagin to be the form, in which the transportation of albu-
minous bodies takes place. The most important investigations,
however, we owe to E. Schulze and his school.

Asparagin has been found normally in many plants, but under
special conditions in still more cases. The root of Althaa contains

1) It can hardly be doubted, that all such substances as are capable of being
converted into starch, as glycerine (Arthur Meyer, Laurent) or glycol or methyl alcohol
(Th. Bokorny) also serve as sources of carbon for the formation of protein compounds
Also the combination of formic aldehyde with primary sodium sulfite may be used
under certain circumstances (Th Bokorny).

56 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

2%, that of Glycirvrliza 0,8% (Plisson), that of Scorzonera 0,6%
(Gorup), Potatoes 3% (E. Schulze). It was found in the root of
Symphytum, in Lactuca, in the shoots of Asparagus, of Humulus,
and of Bambusa (Kozat)*) in the leaves of Atropa” and in the
leaf-buds of Ulimus effusa, Spiraea sorbtfolia, Sp.opulifolia, Populus
tremula, Quercus pedunculata, Lonicera tatarica, Tilia parvifolia,
Alnus (Borodin). Buds of Betula and of different conifers show
normally no asparagin, but soon after the cut twigs are placed in
water. Borodin found that, under this condition, also flowers,
stems and parts of fruits can form asparagin,?) and E. Schulze
found the same with twigs of Fagus sylvatica, Populus nigra, Vitis
vinifera, Acer, Platanus; Betula formed after 10 days 2,0% as-
paragin. Kisser demonstrated that this production of asparagin
is connectedwith a decrease of protein-matter.

Kellner observed, that in young grass often more than 30%
of the nitrogen is present in form of amido-compounds, especially
as asparagin and glutamin.®*) According to Emmerling young
leaves of Vicia are richer in asparagin than old ones, and shoots,
buds and newly formed fruits contain sometimes considerable
quantities. Fresh stems of Medicago sativa contain 7 times as
much asparagin as the fresh leaves (0,05%). The inner bark
(liber) of Platanus contains asparagin, but not that of Quercus,
Tilia, or Fraxinus (E. Schulze).

Boussingault was the first, who found asparagin as a constant
product in plants that are deprived of light.”) Oats, cultivated in
pots, if deprived of light for 7 days, yielded 1,67% asparagin of
the dry matter, the nitrogen of which corresponds to 60% of the
nitrogen of the decomposed protein compounds. Red clover
produced after 8 days in the dark 14 times as much asparagin
as under normal conditions.’ The absence of light brings on a
gradual decrease of carbohydrates, respiration going on and

1) Bulletin Vol. I, No.7 of the Agricultural Dep. of the Imp. University of
Tokio.

2) Husemann and Hilger, Pflanzenchemie Vol. I.

3) Botan. Zeitg. 1878, p. 208.

4) Landw. Jahrb. 1888, p. 702.

5) Ibid. 1879, p, 243.

6) Landw. Versuchsstat. 24, 113.

7) Compt. rend. 58, 881 a. 917.

8) E. Schulze, Landw. Versuchsstat. 36. According to O. Miilley asparagin is
also formed if the growing parts alone are kept deprived of light (Ibid. 33, 310).

THE FORMATION OF PROTEIDS IN PLANT-CELLS, 57

new production being stopped. ‘There appears in many cases a
close connection existing between the decrease of carbohydrates
and an incipient decomposition of protein compounds’ with
production of asparagin. ‘Thus Monteverde has observed, that
twigs of Syrvinga vulgaris, kept in the dark for 15 days, produce
a great deal of asparagin; but if the twigs instead of being
placed in water are kept in a 6-8 per cent solution of glucose or
sucrose, no trace of asparagin is formed, but much starch and
mannite.’) Other investigations have elucidated the interesting
fact, that the amount of asparagin formed in the germination
process increases if the reserve carbohydrates decrease in quantity.
The proportion of protein to carbohyrates is
in wheat-kernels=1: 5

in peas 18
in beans It Sat pi5)
in gourd-seeds =1:1,5
in soya-beans =1:0,9
in lupin-seeds =1:0,5.
Germinating peas contain 2,4-2,6% asparagin of the dry
substance (Sachsse aud Kormann). In the shoots of Cucurbita

(gourd) there are 40% of the nitrogen of the decomposed proteids
present in form of asparagin and glutamin; generally more of
the latter than of the former (EZ. Schulze).?> In germinating
soya-beans half of the nitrogen of the decomposed protein is
found again as asparagin. In the Jupin-seeds, however, the
greatest amount of asparagin is found; the percentage of nitrogen
of the decomposed protein compounds (mostly conglutin), which
is present in form of aspavagin, is with germs 4 days old=45,7
We op ” =56,7
Zs ”» = 515
2AwE I ei) = 7394:
In the axial parts (root and stem) of lupin-shoots the
asparagin amounts to 30% of the dry matter, in the cotyledons
to 8-9%.°) The great amount of asparagin in the stem of a

1) Botan. Centralbl. 1891, 380. Also an observation of Church (Journ. Chem.
Soc. 49, 840) may be mentioned here, according to which leaves suffering from
albinism contain 2,4 times as much nitrogen in form of amido-compounds (which ?)
as green leaves of the same trees (Elacagnus pungens).

2) Landw. Jahrb. Vol. 12 p. 916.

3) E. Schulze, Landw. Jahrb. 17; Journ. f. prakt. Chem. 1883.

58 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

germinating lupin can be well demonstrated by treating a thin
cut through the stem with alcohol under the microscope, a large
number of asparagin crystals will soon be observed.

There exist, however, cases in which a considerable amount
of asparagin is found in presence of a large amount of carbo-
hydrates, as in the shoots of Cannabis sativa and of Helianthus
annuus. In the latter case were found 4% asparagin and 14,7%
sucrose.") Scheibler found in the sugar-beet considerable quanti-
ties of asparagin; more frequently, however, the next higher
homologue, glutamin, in larger quantities than the former.”
In the juice of potatoes freed from albumen, more than 46% of
the nitrogen is present in form of asparagin, although there is
not only a great amount of starch, but also some reducing sugar
present (EZ. Schulze). In Trifolium, Medicago and Vicia is found
1-2% asparagin in presence of 1,5-2% of glucose (E. Schulze).

These numerous facts doubtless reveal a certain physiolo-
gical significance of asparagin. Let us now take a glance at the
other nitrogenous compounds formed by decomposition of
protein bodies. Small quantities of leucin and tyrosin are en-
countered in potatoes, but whether they are formed im loco or
had been transported from the leaves to the bulbs is not decided;
they are however probably decomposition products of albuminous
compounds. The same amido-acids were found by E. Schulze)
in small quantities in germinating seeds of Cucurbita. Kozat
found tyrosin in bamboo-shoots. Tyrosin was found only in
traces in lupin-shoots, leucin however not at all. Phenyl-
amido-propionicacid and amidovalerianic acid occur here
in larger quantities. Also an interesting new base, arginin,
C;H,,N,0. was discovered by E. Schulze and Steiger in the
cotyledons of germinating lupin-seeds. These authors proved
also that this base is derived from the decomposition of proteids ;
it amounted to 7,8% of the dry substance of the cotyledons and
was not found in the axial organs. It is probably derived from

1) Frankfurt, Landw. Versuch-Stat. 43, 143.

2) Ber. d. Deutsch. Chem. Ges. 1869. Schulze and Urich, Landw. Versuchs-Stat,
20, 193.

3) Landw. Jahrb. Vol. 9; Vol. 12; Vol. rq.

4) Zeitsch. f. physiol. Chem. Vol. rz p. 43, Ber. Deutsch. Chem. Ges. Vol.

LO; ps L775

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 59

the same atomic group, that leads by the decomposition of
proteids with hydrochloric acid to lysin C;H,,N.O. and lysatin
C>H,;N;,0., which were discovered by Drechsel. Arginin is con-
tained also in the Cucurbita-shoots but only in very small
quantities ; also in the soya-shoots it appears to be present.’

Another nitrogenous substance, not found hitherto in plants,
was discovered by E. Schulze and I. Barbiert in the young leaves
of Platanus, allantoin.”

Also the bark of Aesculus Hypocastanum and leaves of Acer
pseudo-platanus and Acer campestve contain small quantities.%)
While the shoots of Platanus-buds contain as much as 1%
allantoin, it could not be discovered in many other plants (Vzcva,
Trifolium, Betula, Fagus, Tilia, Populus, Vitis vinifera, shoots of
Cucurbita and Lupinus.)

Urea as such has not yet been found in plants, but the
closely related guanidin was discovered by FE. Schulze in shoots
of Vicia sativa.) Still another base was found by E. Schulze and
E. Bosshard in young Vicia and Trifolium, in cotyledons of ger-
minating Cucurbita and in ergot (Claviceps purpurea); it was called
vermin, corresponds to the formula C,,H,..Ns0;+3H.O and yields
by decomposition with hydrochloric acid guanin.® Later the
same substance was found by E. Schulze and A. v. Planta also in
the pollen of Corylus avellana and of Pinus sylvestris.”) Whether
this base results from decomposition of protein has not been
proved positively, but as to the other products mentioned, there
cannot exist any doubt. Gorup-Besanez, who first discovered
leucin in plants (shoots of Vicia), was also the first to demonstrate

1) E. Schulze, Zeitsch. f. physiol. Chem. 11, 43 and Journ. f. prakt. Chem. 32,
433:

2) Ber. d. Deutsch. Chem, Ges. 13, 1602. Allantoin, CyHe@N403, occurs in the
animal body and is a derivative of urea, a diureid of glyoxylic acid. Also arginin is
a derivative of urea (Schulze) like lysatin (Drechsel).

3) Zeitschr. f. physiol. Chem. 9, 420. Richardson and Crampton found small
quantities of allantoin in germinating wheat (Ber. Deutsch. Chem. Ges. 19).

4) All these plants were kept for some time in the dark to bring on a decomposi-
tion of protein (EZ. Schulze).

5) Ber. d. Deutsch. Chem. Ges. 25, 658. 3 kilo of dry shoots yielded only rg.
of nitrate of guanidin.

6) Zeitschr. f, physiol. Chem. 10, 80.

7) Ibid. 10, 326. From 1300 g. of Corylus-pollen about 1g. of vernin was ob-
tained.

60 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

the presence of a digesting ferment, resembling trypsin, in plants,
to which he ascribed the production of amido-acids from proteids.”

If we compare now the quantities of the different amido-
acids and bases with that of asparagin we find generally the
latter present in larger quantity, although the decomposition of
proteids by enzymes or by mineral acids yields only relatively small
quantities of aspartic acid (which in the plant-cells might easily
be transformed into asparagin). While roo parts of conglutin
(the principal reserve protein in lupins) yield upon decomposition
with hydrochloric acid 6 parts of glutaminic acid, 1,5 parts of
aspartic acid, 10 parts of leucin and 2 parts of tyrosin, we find in
lupin-shoots 12 days old, the asparagin in dominating quantities,
amounting to almost 30 per cent of the dry substance, while
tyrosin was found only in traces and glutamin could not be found
with certainty. Instead of leucin the next lower homologue
(perhaps produced from the former by oxidation), was found.”
The germinating seeds of Cucurbita contain only a very small
amount of leucin, but 1,75 per cent of glutamin and 0,06 per cent
of asparagin; tyrosin amounted here to 0,25 per cent.

According to Ruitthausen gluten-casein yields on decompo-
sition with hydrochloric acid 0,3 % aspartic and 5,3% glutaminic
acid; legumin yields 3,5% of the former and 1,5% of the latter
(only mucedin yields larger quantities of glutaminic acid,
viz. 25%). Still- we find in plants generally asparagin as
the main product,?) the other amido-compounds disappearing
rapidly again and being found only in relatively small quantities.
Schulze supposed that the decomposition of protein in plants
would yield the same products and in the same quantities as
the decomposition by trypsin or by acids, but that the rege-
neration of proteids would proceed more easily from the
other amido-compounds than from aspartic acid, hence the
latter—transformed into asparagin—must accumulate.*) This is,

1) Ber. Deutsch. Chem. Ges. Vol. 7 and ro.

2) E. Schulze and Barbieri, Landw. Jahrb. 1880, p. 18. Most proteids yield by
decomposition with acids more than 20 per cent leucin. ;

3) In certain cases mentioned above, asparagin is fuund replaced by its next
homologue, glutamin. This might he gradually transformed into the former in the
plant-cells or like asparagin serve in the regeneration of certain proteids.

4) Other views upon this subject were discussed by E. Schulze in Landw. Jahrb,
Vol. 17 and 21.—He showed that they are either incorrect or imperfect.

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 61

however, an erroneous assumption, for experiments with bacteria
and with mould-fungi have convinced us that asparagin and
aspartic acid form most excellent nutrients, being evidently very
favorable sources of carbon and nitrogen for the formation of
protein, for the growth of protoplasm, i. e. multiplication of
cells. These combinations are far superior to tyrosin or phenyl-
amidopropionic acid, which would, moreover, have to undergo a
thorough chemical change before the formation of proteids
could commence.

An observation of E. Schulze is, in this connection, of fun-
damental importance. He found that the quantity of amido-
acids formed during the first period of germination is con-
tinually decreasing, whilst the amount of asparagin is zucreasing ;
he showed furthermore, that the proportion of asparagin is much
greater in the axial organs than in the cotyledons, as is seen
from the following table:

The protein-free extract contains

nitrogen in form of:

Lupin shoots.

In the cotyledons. In the axial parts.

-_ |Other amido- .. |Other amido-

BBE A=AE iD. compounds. A ITEMETEI compounds.
() CESS Cel po» 6g co 20,5 7935 68,8 3152
TZNCAYSNOl\eney) lle) wie 26,2 738 78,1 21,1

100 parts of lupin-shoots containing 16,8 parts of asparagin
yielded, after 3 weeks vegetation in diffused daylight, 151 parts
of green plants with 23,9 parts of asparagin.’) The primary
amido-products disappear first, their carbon serves partially to
support respiration, while another part of their carbon together
with their nitrogen is found now in form of asparagin, and this
again disappears finally with the increase of available glucose,
formed by the function of the chlorophyll. The asparagin
evidently indicates the manner of the protein formation, it is

1) Landw. Jahrb. 9, 41. The sulfur of the decomposed protein is converted into
sulfates, which later serve afterwards again in reconstruction of the proteids
(E. Schulze ; Tamann).

62 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

a transitory product, found when the conditions of finishing the
process of protein-synthesis are not complete. ‘To these con-
clusions we are led by the circumstances under which asparagin
appears in shoots, and disappears again.

When sugar participates in the formation of proteids from
asparagin, it evidently yields the carbon to make up the deficiency ;
this function, however, would not be required if aspartic acid
were used; the proportion of the number of atoms of

N: C is in albumen I34
in aspartic acid 134
in asparagin ar 22

(in tyrosin 1:9, in leucin 1: 6).

Several considerations (see Chapt. VIII) make it highly
probable, that the formation of protein compounds consists in a
rapidly proceeding condensation process, in a certain degree
analogous to the formation of sugars from formic aldehyde.”
To render, however, such a process possible, asparagin would
best be converted into the di-aldehyde of aspartic acid, a reduc-
ing process whereby glucose very probably would have to furnish
the necessary hydrogen:

CONH, COH

| |

CH, CH,

l Senet = | +NH,+H.0.
CH.NH, CH.NH,

| |

COOH COH

Asparagin. Aspartic aldehyde.

The conditions in the living cells would not permit for a
moment these products of reductions in a free state, and in
presence of glucose the ammonia set free would be transformed
at once into organic combinations, most probably into another
molecule of aspartic aldehyde; thus asparagin and glucose
would yield directly 2 mol. of this aldehyde, which may be
expressed by the following equation :

1) I have been the first to show that formic aldehyde will by condensation yield
true sugars, and the first to prove beyond a doubt, that such a synthetical sugar is
capable of alcoholic fermentation. No sophistry can disprove these facts.
Compare Journ. f. prakt. Chem. 33, 332; Ber. d. Deutsch. Chem. Ges. 20, 142 and
3042; Ibid. 22, 477 and 481; Landw. Versuchstat. 41, 132.

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 63

C,HsN.0,+CsH,,06+20=2C,H,NO,+2C0O,+3H.0.

The condensation process, however, leading from this
highly labil’)—and still hypothetical—amido-aldehyde to the
active albumen must be accompanied by a reducing process and

by the entrance of sulfur. We may express this process by
the following equations:
3C, H, NO, = (Craigs Ne O,+2 j8l,©)2
———
Aspartic aldehyde Intermediate product

6C,. H,, N,0,+12H+H2S=C,, Hire Nig SO2. +2H, 0.
Lieberkiihn’s albumin-formula,

For the reduction indicated in the latter equation the
presence of glucose would be again required.) If we now make
the assumption that under certain chemical conditions the
aldehyde groups are prevented from acting during the condensa-
tion process upon the amido-groups (which is to be expected
under normal conditions) and that the hydrogen serving for
reduction would transform 12 aldehyde groups into secondary
alcoholic groups (CH OH), causing thereby a pinakon-like link-
ing—then we should have in the final product—the active
albumen-—a substance of extraordinary lability, containing 12
aldehyde- and 18 amido-groups in one molecule, and changing
easily into another product with the loss of its aldehydic
character—the passive albumen.

The accumulation of asparagin is evidently connected with
the gradual disappearance of the amido-products, directly result-
ing from a decomposition of protein. This fact finds its most
natural explanation if we recall the conclusions, to which we
were led by the study of the nourishment of the lower fungi (see
page 49). We had logically concluded that if different com-
pounds serve to yield the same protein, then they must be trans-
formed first into one and the same atomic group from which the
protein formation can start. We had seen that this group cannot

t) Amido-aldehydes are as we have explained in Chapt. III exceedingly unstable
compounds, which may play important physiological rdles in more than one respect.
Wolffenstein observed recently an easy transformation of amido-valeraldehyde into
piperidin, and holds it highly probable that amido-aldehydes form the connecting
links between the fatty series and the alkaloids in plants,

2) On glucose as a reducing agent in neutral or even acid solutions, see Chapt. VI.

64 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

be any other one than form-aldehyde. If the fungi utilise once
leucin, another time tyrosin, a third time tartrate of ammonia,
then an oxidation must set. in to reach the common starting
point.) And this must hold good also for the higher plants, for
it can hardly be assumed that the mode of protein formation is
here entirely different. If for the purposes of transportation and
transformation (conglutin into active albumin and living pro-
toplasm), in lupin-shoots, the reserve protein is first dissolved,
and split into a series of amido-products (leucin, admido-valeri-
anic acid, tyrosin, phenyl amido-propionic acid, arginin, etc.) by
an enzyme, then in all probability and logically all those different
products must be transformed into the common starting group:
formic aldehyde, and their nitrogen be liberated as ammonia;
thus the decomposition of leucin by oxidation might be expressed
by the following equation :

C.H,,NO,+70=2C0.+H,0+4CH,0+NH,.

Form-aldehyde and ammonia, however, act noxiously and do
not remain as such for a second ; aspartic aldehyde being formed.”
This product however does neither remain unchanged, it will yield
either directly active albumen when all conditions are fulfilled, or
it will be stored up as asparagin if not all conditions are united
for protein production.°*)

The asparagin in plants has two sources; it may either be
formed directly from glucose, ammonia (or nitrates) and sulfates,
or it may be a transitory product between protein-decomposition
and reconstruction from the fragments. In both cases the am-
mediate processes connected with the formation of asparagin have
the greatest resemblance or are even identical,—although the
original materials are far different.

1) Compare Chapt. III and Chapt. VIII of this essay. I developed the outlines
of this hypothesis first in Piifg. Arch. 1880.

2) The still hypothetical formation of aspartic aldehyde, from formic aldehyde
and ammonia may be expressed by 4CH20+NH3=C4H7NO2+2H20

3) Closely related to asparagin is succinic acid, found not only often in the
higher plants, but also encountered in fungi. I found this acid also in algae
(Spirogyra), in hay of meadows, in the cambial sap of conifers and in the common
yeast (Ber. Bayr. Akad, d. Wiss. 1878; Journ. f. prakt. Chem. 36). I observed
also this acid as a product of oxidation of albumen by potassium permanganate
(Journ. prakt. Chem. 31, 152).

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 65

To effect the synthesis of proteids,—especially in the
last phases—a certain amount of energy is required. This
energy is procured by respiration. Respiration, however, also
brings on the partial oxidations necessary for transforming
glucose into asparagin or aspartic aldehyde. Where respira-
tion is impeded, the protein production will be retarded. It
is, therefore, not surprising that in the interior of potatoes
and beets, asparagin is found in company with starch and
reducing sugar and that the stem of plants contains more as-
paragin than the Jeaves, as Schulze has found. The stalks
of Medicago sativa contain 7 times as much asparagin as the
leaves. Stems and cotyledons of young lupin-plants cultivated
first in the dark and afterwards 4 weeks in daylight, contained
18,4% asparagin, the young leaves, however, only 6%. The
structure of the leaves and their numerous stomata certainly
secure a much more energetic respiration than the structure
of stems, roots and bulbs. As respiration is best supported by
carbohydrates, it is clear that glucose plays a still more im-
portant rdle in protein formation: it yields chemical energy, and
as the /eaves produce by assimilation of carbonic acid a large
amount of glucose, it follows that the /eaves must be the most
favorable organs of the plants for the production of proteids.
The sun-rays are thus indirectly a great supporter of protein-
formation. Dzrectly, however, no such influence is required,
as I have shown in experimenting with mould-fungi grown in
the dark and in diffused day-light; not only was the develop-
ment of the fungus just as energetic in the dark as in the
light but in some cases it even exceeded the latter. Here
glucose and glycerin served as organic nutrients.’)

But while access of air is indispensable for the production
of asparagin and proteids, such is not the case for the action
of the enzymes in peptonising and decomposing the reserve
proteids in the germinating seeds. Palladin has proved that
shoots,—which remain alive in the absence of air for 24 hours—
cease to produce asparagin under this condition,” while the
production of amido-acids by enzyme is still going on.

Glucose is in more than one respect highly important for

1) O. Loew, Biol. Centralbl. 10, 584.
2) Ber. d. Deutsch. Botan, Ges. 6, 205 and 269.

66 THE FORMATION OF PROTEIDS IN PLANT-CELLS.

protein formation, it is not only a very suitable source of carbon
but also enables reductions, and finally yields the necessary
energy by supporting respiration.

The utility of glucose (and other carbohydrates) is, however,
in connection with the protein still greater than mentioned. It
protects—in the fully developed plant at least—the protein
against decomposition. It seems that the action of the proteoly-
tic enzyme sets in here with the gradual disappearance of the
glucose; as for instance in the case of keeping plants in the dark.
If the respiration process finds for support neither fat nor sugar,
then the reserve protein is attacked and the amido-acids yield a
portion of their carbon for the wants of respiration. Another
portion of their carbon is transformed into form-aldehyde, their
nitrogen into ammonia, and from these two very reactive com-
pounds the innocuous asparagin is formed and stored up for later
use. The reserve protein supports here respiration and plays
therefore a réle different from the case of the development
of a shoot or of leaf-buds, where the tvansportation of nitrogen is
the main object, where a splitting must take place to produce
compounds capable of osmosis.”

In many cases this reserve protein is in full grown plants
only present in form of solution in the vacuole, and may be there
as active or as passive albumen, the latter being easily produced
from the former, for instance by dilute acids, perhaps also by
enzymes. That the active albumen stands in close connection
with asparagin becomes evident in all those cases in which Boro-
din observed the production of asparagin during the development
of leaf-buds; here this depends upon a decrease of the amount of
active albumen stored up in the bark of the twigs, as I have
ascertained by the coffein reaction (see Chapt. IV). This
decrease is also observed if branches of Fagus, Quercus, or Betula
are kept in the dark.”

That the passive albumen is not formed by direct synthesis

1) The supposition of several authors that it is the living protoplasm itself,
that is constantly dissociated into nitrogenous and non-nitrogenous material, the
latter serving for respiration, the former being reconstructed to proteids is simply
absurd, entirely unchemical and wholly unphysiological (see Chapt. 1]).

2) This however does not exclude that the passive albumen present, serves the
same purpose; moreover the active albumen is passing at first into the passive state
before being decomposed into different amido-products,

THE FORMATION OF PROTEIDS IN PLANT-CELLS. 67

but is the product of the transformation’) of the directly formed
active unstable albumen is plainly logical and needs no further
explanation to any one acquainted with the progress of chem-
istry; it is no ‘‘doctrinary assumption,’”’ as a botanist in Ger-
many has supposed it to be.

The development and perfection of the sciences causes a
growing division of labor but the increased study of details often
impedes an insight into the connection of the phenomena of relat-
ed branches of science. A catalogue of isolated facts, however,
accumulated with infinite pains by scientific workers, will have a
still fuller significance by the establishment of a unifying con-
ception. The theory here developed combines and compares
observations on the lowest as well as the highest forms of vegeta-
tion, gives a very plausible account of the mode of protein forma-
tion, leads at the same time to a very natural explanation of the
chemical difference between living and dead protoplasm, and
points to conclusions as to the nature of poisonous actions, which
have been verified by experiments.”

Still, new theories are making their way very slowly. The
history of opinion, says R. Beeton, has three stages. The first
stage is that in which men say it is not true; the second is that
in which they say, there may be something in it, and the third
stage is that in which they say they have been of that opinion
all along! The theory of the active albumen has now reached
the second stage.

1) Compare pag. 31. Chapt. IV.
2) Compare Chapt. III of this essay.

On the Vegetable Cheese, Natto.
BY

K. Yabe, Néogakushi.

Since remote times there has been prepared in Japan from
soya beans, a sort of vegetable cheese called natto. The beans
are first boiled in water for five hours to render them exceedingly
soft. The still hot mass is in small portions wrapped in straw
and the bundles thus formed, well tied at both ends, are then
placed in a cellar, in the middle of which a fire is kindled,
whereupon the cellar is well closed. The heat is left to act
for twenty-four hours, after which the product is ready for con-
sumption. Although the moderate heat of the cellar acts only
for twenty-four hours, there is still a considerable bacterial
change going on. The microbes may be derived either from
the air or from the straw. Of course it can not be expected
that bacteria on the surface of the soya beans would still be
very active. They are probably killed by the five hour’s
boiling.) This product has a peculiar but not putrid smell.
The soft mass of the beans is kept together by a very thick
viscid substance. In this substance I have found four kinds of
microbes present, and the chemical decomposition of proteids
must be due to one or more of these microbes.

1, The Microbes of Natto.

A trace of the viscid liquid of the cheese was inoculated in
a gelatine solution and a plate culture was prepared. After a
few days numerous colonies (1260) appeared, of which four
different kinds could be observed, and of these were prepared
pure cultures. Three of these different cultures were formed by
micrococci, and one by a small, not motile, bacillus liquifying

1) Exceptional cases where bacteria can stand boiling heat still longer are
known, for instance with Bacillus subtilis.

ON THE VEGETABLE CHEESE, NATTTO. 69

gelatine and producing a greenish fluorescence. It forms
white flocculent masses on potatoes, and on soya beans it
forms white colonies. With regard to the micrococci the
colonies of the three kinds can be distinguished by their colors:
yellow, orange yellow, and white.”

The yellow micrococcus belongs to the larger kinds. In
gelatine it forms white colonies along the canal. At the head
of the canal a concavity is formed. ‘The gelatine is in a small
degree liquified. On agar, potatoes, and soya beans, it first
forms white colonies that gradually turn yellowish. On soya
beans it develops the characteristic smell which is observed
with natto itself. In 2% peptone solution it forms a white
deposit.

The orange yellow micrococcus forms round colonies on
gelatine. It has a general resemblance to the former but it does
not liquify gelatine at all, and produces on soya beans a dis-
agreeable smell. While the former does not spread considerably
on potatoes, this one does so forming a slimy cover.

The white micrococcus has a general resemblance, in regard
to growth and development of its colonies, to the last named.
With regard to the specific smell of natto, repeated experiments
have convinced me that the above mentioned yellow micrococcus
is the chief cause, while with regard to the slimy substance
which shows an enormous degree of viscidity further experiments
have to be carried out; because the yellow micrococcus is not
the cause of this viscidity.?)

2, Chemical Investigation.

As the soya bean is very rich in protein, the various decom-
position products of proteids might be expected to be present
in this cheese to a certain extent. 6.8 kilos. of the raw cheese
were extracted with boiling water, and the aqueous solution
precipitated with basic acetate of lead. The filtrate from this

1) White colonies were far less numerous than the others.

2) I made also several experiments to observe the degree of resistance to
hydrochloric acid, and found that the microbes mentioned remained alive in 2°/,
peptone solution containing 0,11 °/, HCl, but they were killed when hydrochloric
acid was increased to 0,19°/..

70 ON THE VEGETABLE CHEESE, NATTO.

precipitate was mixed with mercuric nitrate with the gradual
addition of small quantities of soda as long as a precipitate was
formed. This precipitate, after being washed well on a filter, was
decomposed by sulphuretted hydrogen, and the filtrate evaporat-
ed on the water bath, ammonia being, from time to time, added
to keep the solution neutral. In the concentrated liquid were
formed after some time white crystalline masses composed of
radiating needles of the forms characteristic of tyrosin. They
were easily soluble in dilute ammonia and in hydrochloric acid,
slightly soluble in cold, and easily in hot, water. They were
purified by repeated recrystallisation, and then yielded the reac-
tion of Pirvia, Wurster and Hofmann for tyrosin. The determina-
tion of nitrogen by KAyjeldahl’s method yielded 7,98 %, while the
theory requires 7,75 %. Also the copper compound was ob-
tained by boiling the solution with copper hydrate and filtering
while hot. The total quantity obtained amounted to 3.212
grm. The mother liquor from which the tyrosin was separated
was, farther concentrated and divided into two parts (a) and (b).
The part (a) was precipitated with phosphotungstic acid after the
addition of a little sulphuric acid (c) and the filtrate mixed with
caustic baryta to separate the sulphuric acid and phosphotung-
stic acid. After the removal of the excess of baryta by a current
of carbon dioxide, the filtrate was evaporated. Numerous
spherocrystals were obtained with the behavior of leucin mixed
with the crystals of ammonium nitrate. To remove the latter,
a little baryta was added and by evaporation the ammonia was
expelled. When the residue was treated with alcohol, barium
nitrate remained behind while the alcoholic solution on evapora-
tion yielded crystals of leucin which were converted into the
characteristic copper compound which yielded on analysis
19.76% copper, while the formula (Cs H;; NO.). Cu requires 19,5%.

The phosphotungstic precipitate (c) was first washed with
cold water containing some sulphuric acid and then decomposed
in the usual way with caustic baryta and the filtrate after temo-
val of the excess of baryta evaporated, whereby a syrupy liquid
was obtained. I searched here for lysin, lysatinin (both discovered
by Drechsel) and arginin (discovered by Schulze), which are the
decomposition products of the proteids of the germinating lupines,
but all my efforts were in vain. The syrup gave, however, all

ON THE VEGETABLE CHEESE, NATTO. 71

reactions of peptone and consisted for the most part of this
substance.

The above mentioned part (b) was treated with ammo-
niacal solution of silver nitrate whereby a small amount of a
white precipitate was obtained which was collected on a filter
and washed with dilute ammoniacal solution of silver nitrate,
then dissolved in warm nitric acid of sp. gr. 1,1 with the addition
of little urea. Upon cooling, microscopical needles were obtain-
ed, which proved to be a mixture of the silver compound of
guanine and of hypoxanthin. After the removal of the silver
with sulphuretted hydrogen, filtering and evaporating with the
addition of a little ammonia, a residue was obtained, soluble
with great difficulty in water and alcohol but easily soluble in
mineral acids. It was treated with ammonia whereby a part
was dissolved and a part not. The latter gave the sharp reac-
tion of Capranica for guanine. When dried with nitric acid in a
platinum dish it gave a yellow residue, which turned red on the
addition of soda. The former, i. e., the soluble part was ob-
tained by evaporationg the ammoniacal liquor. When evaporat-
ed in a platinum dish with nitric acid, and the residue treated
with caustic potash, no coloration took place. The reaction
of Wedel and of Capranica did not leave any doubt that this
substance was hypoxanthin, but there was contained also xanthin
in the cheese. This was obtained by adding ammonia to the
filtrate separated from the first crystallisation of the guanine
and hypoxanthin silver compounds. By adding ammonia, a
yellowish flocculent precipitate was obtained from which the
silver was removed by sulphuretted hydrogen. The filtrate then
evaporated to dryness left a faintly yellowish powder slightly
soluble in water, insoluble in alcohol and ether, but easily soluble
in alkalies and acids. On treating it with nitric acid a yellow
residue was obtained turning red upon the addition of soda
and purple on heating. The reaction of Hoppe-Seyler and
Wetdel left no doubt that this substance was xanthin. Wheth-
er these substances of the xanthin series were formed by the
bacterial action during twenty-four hours in the warmed cellar
is doubtful. I think it is more probable that they were origi-
nally present in the soya bean. But there can be no doubt
that a large portion of peptone, and also leucin and tyrosin

72, ON THE VEGETABLE CHEESE, NATTO.

were products of the bacterial action. Considering the high
temperature of the warmed cellar, the considerable extent of
the bacterial decomposition is not very surprising. There can
hardly be any doubt that the matto-preparation is more easily
digestible than the original soya bean, as it is very soft”
and contains more peptone.

We add finally the determination of the different forms of
nitrogen in the soya been and in natto.

Soya Natto prepared
bean. from the same
soya bean.
otal nitrogen.” 75355 % 7,542 %
Nitrogen of proteids
(excluding peptone) 6,899 4,033
Nitrogen of amides 0,128 1,892
Nitrogen of peptone 0,328 T5007,

1) While the water of the air-dry soya bean, amounted to 15,16 0/o, that of natto
amounted to 59,120/o.

2) The relative increase of total nitrogen in natto may be chiefly due to the
loss of carbon as carbon dioxide during the fermentation.

On the Poisonous Action of the Hydroxyl-derivatives of Benzol
upon Yeast and Bacteria.
BY

K. Yabe, Nogakushi.

The toxical action of phenol, of the dioxybenzols (resorcin,
pyrocatechin and hydrochinon) and of the trioxybenzols (phloro-
glucin,” pyrogallol) has been compared in regard to animals but
not yet in regard to the lower fungi. As a general rule, it has
been found that the poisonous character increases with the
number of hydroxyl groups entering into the benzol ring.
Stolnikow observed that phloroglucin is more poisonous for frogs
than resorcin, and this is again more poisonous than phenol.
While the lethal dose of phenol for warm-blooded animals is
0,3-0,7 gr. per kg., 0,08 gr. resorcin is found to be sufficient
(Zent and Betelli). ‘The three isomeric dioxybenzols show a very
great difference in their toxical action. Pyrocatechin is strong-
est, then follows hydrochinon and finally resorcin. Of-the trioxy-
benzols, phloroglucin is less poisonous than pyrogallol. Loew”
observed that in r per mille phenol solution some infusoria still
remained alive after 15 hours, and certain algee have been found
alive after three days, while pyrocatechin solution of the same
strength killed infusoria and diatoms after a few minutes, spiro-
gyra after several hours. Hydrochinon acts somewhat more
slowly, but in resorcin solution of that strength infusoria live
several hours, and alge are found alive even after 18 hours.
The phenol character is here evidently increased by the toxical
effect caused by the capability of absorbing oxygen. Pyrocatech-
in and hydrochinon will rob the cells of the dissolved molecular
oxygen much more quickly than phloroglucin, and consequently
we find with the former also a much more poisonous character.

1) The third isomeride, oxyhydrochinon, has not yet been physiologically
studied.

2) Natiirliches System der Giftwirkungen, p. 50-51.

74 ON THE POISONOUS ACTION OF THE HYDROXYL-DERIVATIVES

It seemed to be of interest to compare these different
phenols in relation to yeast and bacteria as it has been recently
asserted by Buernacki (Pfliig. Arch. 49, 112) that phenol is a
stronger poison for yeast than pyrogallol which would be an
exceptional case, the alcoholic fermentation being prevented by
the following solutions :—

Phenolac sen eee oe oe 2 OO)
RGSOLCIME cee eae LOO
Pyrogallol igen oceans eee va Or

For this purpose, equivalent quantities of different phenols
were dissolved in Pastewr solution, and yeast added. The results
are shown in the following table :—

Phenol 0,5% no fermentation. || 0,4 % no fermentation.

Pyrocatechin 0,585 ,, me 0,468 ,, feeble fermen-
tation.

Resorcin 0,585 fermentation.

Hydrochinon 0,585 Aas SS

Pyrogallol 0,670 x ——

Phloroglucin 0,670 55

In a second experiment, yeast was shaken first with the
solutions of these phenols and left to stand for 70 hours. After
decantation of the liquid, the yeast was put in Pastewr solution
with the following results:

Phenol 0,3 % fermentation. 0,4 % no fermentation.
Pyrocatechin 0,351 Ae 0,468 feeble fermenta-
tion.

Resorcin 0,351 6 0,468 fermentation.
Hydrochinon 0,351 re 0,468 -,
Pyrogallol 0,402 99 0,536 £
Phloroglucin 0,402 3 0,536 25

Phenol 0,45 % no fermentation.

Pyrocatechin 0,527 3

Resorcin 0,527 fermentation.

Hydrochinon 0,527 feeble fermentation.

Pyrogallol 0,603 “5

Phloroglucin 0,603 fermentation.

The same experiment has been carried out with bacteria.
A drop of putrid peptone solution was mixed with 50 c.c. of the

OF BENZOL UPON YEAST AND BACTERIA. 75

equimolecular solutions of phenols and left to stand for 70 hours.
Afterward traces of the different liquids were inoculated in ste-
rilised peptone solution with the following results :—

Phenol Ae es
Pyrocatechin ,468
Resorcin 468

Hydrochinon — ,468
Pyrogallol 9536
Phloroglucin 536

no development.

development.
no development.
development.

99

We observe here, therefore, an exception to the rule, the
higher hydroxylated benzols are in the case of the lower fungi

less poisonous than phenol itself.

Further experiments must be

made to bring out a scientific explanation of this exceptional

behavior.

On the Quantity of Wood-gum (Xylan) contained
in Different Kinds of Wood.

BY

J. Okumura, Nogakushi.

The durability of wood is of great importance for industrial
purposes. This durability depends not only upon the greater or
less density, but also upon the presence of certain chemical cons-
tituents. Thus a certain proportion of resinous matters will
increase the durability, while the presence of easily soluble
carbohydrates may diminish it considerably. The durability
consists in a certain resistance to the attacks of different kinds
of fungi, as Polyporus, Agaricus, etc. Resinous matters can
never be attacked by these fungi, but various carbohydrates,
as starch, wood-gum, and even cellulose are sometimes rapidly
dissolved by the ferments produced by these fungi which thus
prepare their way to penetrate into the interior of the wood."
From this stand-point it seemed to me to be of great interest to
determine the amount of wood-gum in a series of trees grown in
Japan. Of course the quantity of wood-gum will not be found
always to be a constant figure. The proportion may differ
somewhat between the heart-wood and bark, and also may yary
with the age of the tree. It is certainly of physiological interest
that Thomsen found great differences in the amount of wood-gum
in sap-wood and heart-wood as seen from the following table :—

Dried at 100°C. % in dry matter.
Sap-wood. Heart-wood.

Betula alba L. (old) 13.9 19.7

a (young) 24.9 26.4
Fagus sylvatica, L. (100 years old) 8.2 15.9

95 (young) I1.g 11.3
Quercus glandulifera, Bl. 14.4 10.7
Prunus pseudocerasus, L. 19.3 15-4

1) The observations of R. Hartig in this direction are of special importance.

ON THE QUANTITY OF WOOD-GUM (xYLAN) 77

Fraxinus pubinervis, Bl. 9-7 10.7
Ulmus parvifolia, Jacg 8.9 12.0.

The samples which I took for the determination of wood-
gum were collected from the specimens that belong to the
forestry department of our college. 5 grams of the finely
powdered air-dried wood’) were extracted with 50 c.c. of 5%
solution of caustic soda. After standing for 24 hours with
occasional stirring it was filtered and washed with cold water.
The filtrate was then mixed with hydrochloric acid until a weak
acid reaction was perceptible. The flocculent precipitate was
then collected on a weighed filter and dried at 100°C. to con-
stant weight. In this way I obtained the following results :—

Percentages of wood-gum

in the dry matter.
1) Cryptomeria japonica, Don. (Coniferz) 1.742

2) Thuya obtusa, B. et H. Gass! a>) Zea
3) Pinus parviflora, S. et Z. ( a ee) 4.212
4) Ginkgo biloba, L. ( - a) 2.519
5) Pinus thumbergii, parlat. ( re) 4.500
6) Abies firma, S. et Z. ( a, 0.961
7) Torreya nucifera, S. et Z. ( 3 = 4) Pog oa
8) Podocarpusmacrophylla,Don.( ,, ) 2.914
9g) Zelkowa acuminata, Planch. (Ulmaceae) 13.240
10) Castanea vulgaris, Lam. var.
japonica, DC. (Cupuliferae) 4.776
11) Fagus Sieboldi, Endl. ( ie ) 19.716
12) Quercus acuta, th. ( m ) 6.609
13) Alnus incana, wild. var.
glauca, Ait. (Betulaceae) 6.852
14) Phellodendron amurense,
Rupr. (Rutaceae) 6.586

15) Magnolia hypolenca, S. et Z. (Magnoliaceae) 10.327
16) Cladrastis amurensis, B. et H.

var. floribunda, Maxim. (Legminaceae) 11.964
17) Melia azedarach, L. var.

subtripinnata, Mig. (Maliaceae) 2.634
18) Ternstrcemia japonica, th. (Ternstromiaceae) 3.813

1) All of these were sap-wood except that of Cryptomeria japonica.

78 CONTAINED IN DIFFERENT KINDS OF WOOD.

19) Acanthopanax innovans, (Araliaceae) 8.409
3h cia Za

20) Juglans mandshurica, Maxim. (Juglandaceae) 6.985
21) Phyllostachys nigra, Munro. (Gramineae) 6.234
We see, therefore, that the Coniferze are comparatively poor
in wood-gum, and Ternstroemia and Melia are also poor.
Cupuliferee are richer, Juglans, Magnolia, Cladrastis, Acantho-
panax, etc. are still more so.

On the Reserve Protein in Plants.

BY

G. Daikuhara, Négakushi.

While in the seeds the reserve protein forms small globules
called aleuron, it is with fully developed plants in many cases
not yet decided how the reserve protein occurs. It is, however,
generally supposed that albumen is present in solution in all
the plant juices and this albumen may be used up in all
those instances when a considerable amount is wanted at a
certain time, as for instance during the ripening of the seeds.
Recent investigations have shown, however, that the albumen
dissolved in the vacuole is not always the ordinary or passive
albumen, but in many cases a very unstable albuminous
body which is closely related to the albuminous substance
in the living protoplasm, and which has been called active
albumen. This changes its nature soon after the cell dies,
and is turned into the ordinary or passive albumen. In many
cases, however, this active reserve protein in the vacuole
is changed in this way while the cells are still alive. It has
not yet been determined how far this active albumen is con-
nected with the production of amido-compounds which takes
place when the branches of trees are placed in water, or when
entire plants are kept deprived of light, or when leaf-buds de-
velop on the branches in the spring. I have made, therefore, a
series of experiments to obtain some information about the func-
tion of the active albumen which is found in the bark, leaves,
flowers, and roots of numerous trees, as Quercus, Tilia, Paonia,
Fagus, Prunus, etc. This active albumen can be easily found
by treatment with coffein under the microscope. Cells con-
taining it will show at first numerous little globules in the
vacuole, which flow together to one or several large drops,

1) See Loew and Bokorny, Flora 1892; and Biol. Centralbl. 1891.

80 ON THE RESERVE PROTEIN IN PLANTS.

called proteosomes, with a high refractory power for light.”
Upon treatment with iodine solution, they assume a yellow tinge
and lose their brightness entirely, showing either numerous little
vacuoles or one large one and being then hollow spheres. To
distinguish them from starch granules or fat drops is, there-
fore, a very easy matter. Sometimes it suffices to take a little
piece of fresh bark or a leaf and tear it into little particles
in a drop of coffein solution and observe the result at once
under the microscope with a high magnifying power. Thus,
not only the bright drops mentioned can be seen very soon,
but also in some early dying cells the change from the bright
drops to hollow spheres can be observed even without killing
with iodine.

With regard to the behavior of the proteosomes, I observed
the following: the petals of Saxifraga sarmentosa which had
remained in cold saturated coffein solution and showed then
numerous proteosomes, were left for 15 hours partly in 1% HCl,
partly in 1% HNO,, and partly in dilute phosphotungstic acid.
The proteosomes were not dissolved but had become turbid.
Nitric acid had produced a yellowish coloration which became
stronger on heating.

The petals of Punica granatum were, after treatment with
coffein, placed (a) partly in 1 p. mille NH,, (b) partly in 1%
acetic acid and (c) partly in alcohol of 20%. After 4 hours it
was observed that the dilute NH, had not produced any vacuoles
in the proteosomes; it seemed that they had become solid, and
neither absolute alcohol nor NH, of 10% nor acetic acid of
1% changed them any farther. The portion exposed to the
dilute acetic acid (b) showed coagulated masses of irregular
form which were not changed any more by absolute alcohol.
A portion of proteosomes seemed to have been partly dis-
solved. Those proteosomes (c) which had remained 4 hours
in alcohol of 20% were partly transformed to hollow spheres,
partly to irregular masses which experienced no farther change
by treatment with absolute alcohol.

1) Dead cells never give proteosomes on treatment with coffein. If leaves of
Pzonia albiflora, for instance, are left for 24 hours in 1°/, acetic acid or for 1 hour
in vapors of ether, coffein will not produce any more proteosomes while the fresh
leaves give a very strong reaction.

ON THE RESERVE PROTEIN IN PLANTS. 81

By exposure to boiling 5% NaCl solution the proteosomes
were coagulated. This was observed with the petals of Punica
and of Astilbe and the root of Thestum.

Millon’s and biuret reactions are best made on objects free
from tannin, as for instance, the root of Thesium. Both reac-
tions were obtained after fixation of the proteosomes by dilute
ammonia after the modification, described by Loew and Bokorny
(Bot. C. 188g); biuret reaction was also obtained by boiling
with a concentrated solution of copper sulphate solution, washing
and moistening with concentrated solution of caustic potash.

A small branch of Quercus dentata, showing in the young
leaves as well as in the bark, during development in the spring,
a large quantity of active albumen, was left for twelve days with
the stem immersed in water. At the same time two of the
leaves were placed in a small vessel with water. In the latter
case, the respiration was of course restricted to a certain extent
for obvious reasons. Both objects were placed in a corner of a
room with only a moderate amount of light") for twelve days,
and then for two days in the dark. Now the leaves on the
branch commenced to show brown spots, whereupon a micros-
copical examination was made and it was found that there was
no longer any starch in either case, but while the leaves on
the branch no longer showed the reaction of active albumen,
those kept in water still gave a moderate reaction. It might be
objected here that there was just as much albumen still present
in the former leaves, but it was only changed to passive albumen.
But it has been shown by £. Schulze that protein compounds
are transformed in plants kept in darkness into amido-acids, and
that finally asparagin remains as a chief product.

The gradual disappearance of the active albumen from the
leaves stands in close relation to the formation of asparagin;
I found that while the fresh leaves of Quercus glandulifera con-
tained 0.218% of nitrogen in the form of asparagin (determined
after Sachse-Kormann), those kept for seven days in darkness
contained 0.606%; therefore the asparagin was increased near-
ly three times the original amount.

A similar experiment was made with the leaves of Paonia

1) At the lower end of the branch a fresh surface was cut repeatedly in order to
secure the sufficient ascent of water.

82 ON THE RESERVE PROTEIN IN PLANTS.

albiflora, which were kept for 25 days in a large glass jar con-
taining some water in order to secure perfect saturation of the
air with aqueous vapor. The vessel was covered with a glass
plate and exposed to a temperature of 25-30°C. in the dark.
The air was from time to time renewed. After 25 days the
leaves commenced to show black spots and the still healthy
part of the leaves failed in many cells to give proteosomes by
treatment with coffein, while in the beginning an exceedingly
strong reaction, especially in the epidermis-cells, had been ob-
served. The decrease of the amount of active albumen stands
evidently in close relation to the production of asparagin also
in this case where the amount of asparagin was increased con-
siderably as seen from the following result :—

Fresh leaves. Starved leaves.

Total. Ny g:ne0 ween 22227, 2.648 %
Album. No x.¢° seer 1.890 ,, it Ata
Asparagin-N. os... cs 0.250), 1. 20re

Total N: Aspar:No =. 100 : 9.86 100 : 45,72

In order to determine whether the active albumen occurs
very frequently in plants, I made numerous microscopical exami-
nations, the results of which are shown in the following tables.
While a large number of plants contain in different parts active
albumen in the vacuole, other plants contain only passive,
and again others do not store up any albuminous matter at all in
the fully developed tissues. Such parts of plants as grow just as
quickly as the formation of albumen proceeds, will of course not
be able to store up the latter. Other plants may convert, either
by acids or by ferments,’) the active albumen in their vacuole,
as soon as it is formed, into passive. ‘Thus, for instance, the
leaves of Diospyvos contain neither active nor passive albumen,
the leaves of Vicia contain only passive albumen, while the

1) To decide whether a ferment is the cause of this change in some plants, a ~
cold extract of leaves of Vicia Faba was left to act upon small pieces of the leaves
of Paeonia but no decrease of the active albumen in the cells of the latter
was noticed after 24 hours. Of course it may be doubtful whether a ferment
present could have entered easily into the living tissue.

ON THE RESERVE PROTEIN IN PLANTS. 83

leaves of Prunus contain a large amount of active albumen. The
passive albumen may easily be found in leaves that do not
contain active albumen, by crushing them in a mortar, adding
water, filtering, and adding nitric acid to the filtrate. The co-
agulation by this acid or by heat leaves no doubt that albumen
was present. Some biological relations of interest are re-
cognized from the inspection of the following tables :—

ROSACEAE,
SPECIES. OBJECTS TESTED. ACTIVE REMARKS.
ALBUMEN.
Prunus persica. Young leaves. Present. No starch.
Much
Prunus pseudocerasus. Young leaves. present. No starch.
” ” Epidermis of root. Present. No starch.
” ” Inner tissue of root. None. Much starch.
Pyrus japonica. Young leaves. Present. No starch.
60 ay Roots. None.
sr - Much /
Photinia glabra. Young leaves. present, Much starch,
Much
oa a Flowers. present.
Rosa laevigata. Much h.
vigata Young leaves. present. No starch
Much h
” ” Flowers. present. No starch,
Rosa Banksiz. Roots. Present. No starch.
Kerria japonica. Much N ch.
japonica Young leaves. present. o star
Neutral reaction.
Rubus palmatus. Young leaves. None. Neutral reaction.
” ” », berries, None. Acid reaction.
Rubus Thumbergi. Young leaves. None. No starch.
PALMEAE.

Trachycarpus excelsa. Young buds. | None. No starch,

84 ON THE RESERVE PROTEIN IN PLANTS.

CUPULIFERAE.
SPECIES. Oxyects TESTED. | ACTIVE | REMARKS.
Quercus glandulosa. Young leaves. Much No starch.
present.
3 “ Roots. Present. Neutral reaction.
Much
Quercus dentata. Young leaves. present. Much starch.
Quercus acuta Young leaves woos No starch
cs 8 ; present. ‘
Much
Castanea fesca. Young leaves. present. No starch.
Acid reaction (trace).
TERNSTROEMIAOEAE.
Camellia japonica. Young leaves. Present. No starch.
Camellia theafera. Young leaves. Present. Little starch.
» a3 fas Roots. Trace. Little starch.
Eurya japonica. Young leaves. None.
SIMARUBIAE.

; Much : : :
Ailanthus glandulosa. Young leaves. sesent Slightly acid reaction.
Picrasma ailanthoides. | Young leaves. Present.

Melia Azedararch. Young leaves. None. | Much starch.
HAHAMERIDEAE.

Distylium racemosum. Young leaves. None.

Corylopsis pauciflora. Young leaves. Present.

1) The same species grown in the shade of large trees contained neither active
albumen nor starch.

ON THE RESERVE RROTEIN IN PLANTS.

ORCHIDEAE.

ACTIVE
SPECIES. OBJECTS TESTED. Rae: REMARKS.
Bletia Hyacinthina. Young leaves. Present. No starch.
AD Af Flowers. Present. No starch.
“A a Roots. Present. No starch.
a rh Tubers. Present. No starch.
Gastrodia elata. Flowers. See No starch.
Much
” ” Tubers. ese. No starch.
Cynbidium virens. Young leaves. None.
9 a Roots. None.
Iris tectorum. Young leaves. None. No starch.
Much
9 Roots. present, No starch.
RANUNCULACEAE,
- : Much
Pzonia albiflora. Young leaves. present.
Much
” af Flowers. present.
” ” Tubers. Present. Slightly alkaline
reaction.
Pzonia Moutan. Young leaves. Seat No starch.
” ” Young roots. Present. No starch.
Clematis florida. Young leaves. None. No starch.
Flowers. None. Neutral reaction.
MALVANCEAE.

(ca + nn EE ee ee ee ee ee

Hibiscus syriacus.

Young leaves.

None.

Much starch,

Neutral reaction.

a ee eee

86 ON THE RESERVE PROTEIN IN PLANTS.

ULMACEAE.,
A
SPECIES. OBJECTS TESTED, | ,700IN" _ Remarks.
Zelkowa acuminata. Young leaves. Present.
” ” Roots. None,
Ulmus parvifolia. Young leaves. Present. No starch.
ELAEAGINEAE.
Eleagnus pungens. Young leaves. | None. | Little starch.
MAGNOLIACEAE.
Cercidiphyllum Much
japonicum. Young leaves. present. No starch.
Schizandra chinensis. Ay ap None. Much starch.
3 Ay Young seeds. None. Much starch.
LYTHRARIAE.
Punica Granatum. Young leaves. None.
Much
3 x 1) Flowers. present.
ane As Much
Lagerstroemia indica. Young leaves. present.
PAPAVERACEAE,

Papaver somniferum. Young leaves. Much starch,

Flowers. Neutral reaction.

Unripe seeds.

1) Antipyrin acts just as coffein on the active albumen, but its action is much
slower than that of coffein.

ON THE RESERVE PROTEIN IN PLANTS.
MORACEAE,
SPECIES. OBJECTS TESTED. Rha REMARKS,
Morus alba. | Young leaves. | None. No starch,
” ” ” ” Much pass. albumen.
” ” ” ” Neutral reaction.
ARALIACEAE.
Helwingia japonica. Young leaves. Present. Much starch.
» ” » ” Neutral reaction.
Acanthopanax aculeatum.| Young leaves. Present. No starch.
” » Roots. None,
OLEACEAE.
Ligustrum ibota. Young leaves. None.
Syringa vulgaris. Young leaves. None, | Little starch.
” ” ” ” Little pass. albumen.
CAPRIFOLIACEAE,
Viburnum dilatatum, Young leaves. None. No starch.
Sambucus racemosa, None

var. Sieboldiana. onae leaves.

88 ON THE RESERVE PROTEIN IN PLANTS.
POLYGONACEAE.
SPECIES. OBJECTS TESTED. eee REMARKS.
Polygonum cuspidatum. | Young leaves. Present.
Fagopyrum esculentum. | Young leaves. None.
Much
‘ 3 Flowers. present. No starch.
Pr 3 Roots. None. Much starch.
LAURACEAE.
Cinnamomum Camphora.| Young leaves. None. Much starch.
Meee CACIREE NE EN Young leaves. None. No starch.
ajor.
SOLANACEAE.
Physalis Alkekengi. Young leaves. None. Much starch.
es 34 Flowers. None. Much starch.
0 9 Roots. None. Much starch.
Solanum tuberosum. Young leaves. None. Little starch.
op 0 Flowers. None. No starch.
STERCULIACEAE.
Sterculia platanifolia. | Young leaves. Present.
CORONACEAE.
Aucuba japonica. | Young leaves. | “None.

ON THE RESERVE PROTEIN IN PLANTS. §9g

SAXIFRAGEAE.

< ACTIVE
SPECIES. OBJECTS TESTED. AURUMEN. REMARKS.
Much
} a. Young leaves,
Saxifraga sarmentos g macecnt:
= a Flowers. Much
present,
Much
Roots.
ie ui 28 present.
Deutzia scabra. Young leaves. None. No starch.
“6 nA Flowers. None. Much starch.
7s 5 Roots. None.
: 4 ; The proteosomes contract-
Astilbe japonica. Young leaves. Present.| ed here by treatment with
iodine in such a way that
ie Pe Flowers. Present.| a number of radial fissures
were formed.
Hydrangea paniculata. | Young leaves. None.

VERBENACEAE,.

Premna japonica. Young leaves. None. | Little pass. albumen.
a F Flowers. None.
: P : Much
Callicarpa japonica. Young leaves, present.
SANTALACEAE.
Thesium decurrens. Young leaves. None, Little starch.
Much
32 AEE present.
1 % Flowers, Trace.

1) If we make a vertical cut through the surface of the root, we can observe
under the microscope in many cells globular masses consisting of active albumen
which may have been separated from the dissolved albumen by mechanical action
but after treatment with coffein the quantity is considerably increased. There is
present much tannin in the hot water extracts of the stem but not in those of the root.

go

ON THE RESERVE PROTEIN IN PLANTS,

CORYLACEAE,

> A ,
SPECIES. OBJECTS TESTED. § | J. REMARKS.
Carpinus cordata. Young leaves. Present. No starch.
BETULACEAE.
eee : Much
Alnus maritima. | Young leaves. present. No starch.
ALISMACEAE.
Alisma Plantago. | Young leaves. Present. Much starch.
EBENACEAE.
i
Diospyros kaki. Young leaves. None No pass. albumen.
+ ; Flowers. None Neutral reaction.
3 9 _ Seeds. None
{
DIOSCOREAE.

Dioscorea japonica. Young leaves None. No starch
” » » ” Neutral reaction
” ” ” ” Much pass. albumen?),
= 5 | Roots. None. Neutral reaction.
SALICINEAE.
Salix japonica. Young leaves. No starch.

| None.

1) In this root is also contained about 8‘/, of the dry matter mucin, according to

J. Ishii.

See this Bulletin, p. g9.

ON THE RESERVE PROTEIN IN PLANTS. OL

SAPINDACEAE.
SPECIES. OBJECTS TESTED. ake REMARKS.
Acer palmatum. Young leaves. Present.) Also leaves suffering
from albinism gave
” ” Roots. Present. the reaction,
LILIACEAE.
Lilium collosum. Young leaves. None.

5 “ Roots. None.

Hemerocallis flava. Young leaves. None. | Little pass. albumen.
n 5 Roots. None. No starch.

1 t licul-
poeone Gar eanakeu) Young leaves. None. No starch.
Polygonatum canalicul-| Fjowers, None. No starch.

atum.

RUBIACEAE.
Gardenia florida, Young leaves. None. | Much pass. albumen.
” ” ” ” Much slimy matter.
BERBERIDIEAE.
Nandina domestica. Young leaves. None, No starch.
Much
‘p a Buds. present. No starch.
ONAGRACEAE.
Oenothera Jacquinii. Young leaves. eee No starch.
Much
of Flowers. present. No starch.
” " Roots. panels No starch.

present.

g2 ON THE RESERVE PROTEIN IN PLANTS.

SPECIES.

Lychnis floscucculi.

CARYOPHYLLEAE,
OBJECTS TESTED. pe cTIVE
Young leaves. None.
Flowers. None.

REMARKS.

Dianthus superbus. Young leaves. None. Little starch.
Pass. albumen is found
” » Boots: was: very much in the leaves.
GRAMINEAE.
Arundinaria japonica. Young leaves. None. No starch.

. FA, Roots. None. Immense quantity
of starch in the
interior tissue.

Young shoots, None. Much starch.
Neutral reaction.
Epidermis of
Bambusa senanensis. Deena oS Dowe Present, Much starch.
Triticum vulgare. Bee of young | Present. Little starch.
eeds.
Brachypodium Epidermis of young | Much Nuch serene
japonicum. seeds. present.
Epi BES
Hordeum disticum. biden Aba None. Much starch.
Avena sativa. Epidermis of young | None. Much starch.
seeds.
CRUCIFERAE.
Raphanus sativus. Flowers. No starch.

| None.

In the root of almost all species of plants, the epidermis is generally richer in
active albumen than the interior tissue, while the starch is generally wanting in the

epidermis.

Often the active albumen decreases in those leaves that are nearest to the flowers,
if the latter are very rich in active albumen.

ON

SPECIES.

Vicia sativa.

THE RESERVE PROTEIN IN PLANTS. 93

LEGUMINOSAE.

OBJECTS TESTED.

ACTIVE
ALBUMEN.

Young leaves.

Vicia sativa.

Vicia Faba.

Vicia Faba.

Vicia unijuga.

Lespedeza sericea.

” ”

Wistaria chinensis.

None.

REMARKS.

Much pass, albumen.

Much starch.

Neutral reaction.

Much pass. albumen.

Much starch.

Neutral reaction.

Much pass. albumen,

(Neutral reaction).

Ginkgo biloba.

” ”

Little starch.

Citrus fusca,

” ”

No starch.

Much starch.

Xanthoxylum piperitum.| Young leaves.

Orixa japonica.

Little starch.

Little starch.

Much starch.

Epidermis of seeds. None.
Roots. None.
Young leaves. None.
Epidermis of seeds. None.
Roots. None.
Young leaves, None.
Young leaves. None.
Roots. None.
Young leaves. None. | No starch.
.
CONIFERAE.
Young leaves. None.
Roots. None.
RUTACEAE,
Young leaves. None.
Roots. None.
Much
present.
Much
Roots present.
Young leaves. None.

94 ON THE RESERVE PROTEIN IN PLANTS.

COMPOSITAE.

SPECIES. OBJECTS TESTED. pone REMARKS,
Hieracium umbellatum.| Leaves. None. Acid reaction.
Teucane Chry- Young leaves. None. Much starch.
Leucanthemum Chry-

santhemum. Flowers. None. No starch.
Leucanthemum Chry-
santhemum. Roots. None. No starch.
Petasites japonica. Young leaves. None. Much starch.
MENISPERMACEAE.
Cocculus indicus. Young leaves. None.
Menispermum dahuricum.| Leaves. None.
i * Flowers. None.
CONVOLVULACEAE.
Ipomaea hederacea. Young leaves. None. No starch.
* sf Roots. None. Much starch.
ANACARDIACEAE.
Rhus semi-alata, Young leaves. Present. Much starch.
var. Osbeckii. Roots !). Present.
ERICACEAE.
Pieris japonica. Young leaves. Present. No starch.

1) In the root the active albumen is less than in the leaves.

ON THE RESERVE PROTEIN IN PLANTS. 95

UMBELLIFERAE.
SPECIES. OBJECTS TESTED. eee REMARKS,
Daucus carota. Nerves of leaves. Present.
Thorylis anthriscus. Nerves of leaves. Present.
ILICINEAE.
Ilex pedunculosa. Young leaves. Present. No starch.
Flowers. Present. No starch.
LINACEAE.
Reinwardtia trigyna. Young leaves. None. Little starch.
Flowers. None. Little starch.
CELASTINEAE.
Evonymus japonicus, Young leaves. None. Little starch.
Besvee suberie from | None, Little starch.
Elowers. Present. Little starch.

From these tables it will be seen that of 104 species of
plants, 51 contained albumen stored up, in one part or another,
active while 53 did not contain it at all.

The plants examined belonged to 52 families.

The active albumen was found in 2g families.

It is of physiological interest to see how the active albumen
often accumulates in the flowers and that in such cases it may
be absent in the green leaves or decrease in those parts that
are nearest to the flower (compare the above observations with
Pumca Granatum, Nandina domestica, Evonymus japonica and
Fagopyrum esculentum). In Gramineae I found active albumen
thus far only in the epidermis of seeds and only in a certain
period of development.

96 ON THE RESERVE PROTEIN IN PLANTS.

In the shade active albumen is formed in smaller quantities
than in full sun-light; I found much of it in the leaves of
Ailanthus glandulosa in the latter case, but none in shade-plants.
Further, the young leaves are richer in it than old ones.
Leaves suffering from albinism show it in the white parts of
their leaves about just as much as in the green parts (observa-
tion with Acer palmatum).

On the Occurrence of Mucin in Plants:
BY

J- Ishii, Nogakushi.

In the animal as well as in the vegetable kingdom, there are
found substances of slimy nature, which serve for different
physiological functions; but while the slimes of animals belong
to the protein compounds, all plant-slimes consist, so far as is
yet known, of carbohydrates, corresponding either to the formula
CsH,.O, or C,,H,,0,, and yielding by hydrolysis different
kinds of sugar; as, for instance, carrageen or the slime of an alga
Chondrus crispus Lyngb) yields galactose (Hddicke, Bauer and
Tollens), cerasin or the gum of the cherry-tree yields arabinose
(Scheibler, Kiliant), and the slime of beer-yeast yields mannose
(Hessenland).

As a number of Japanese plants are so rich in slimy matters
as to be applied on account of this quality for industrial pur-
poses, I believed it to be, from a physiological as well as a technical
point of view, of some interest to investigate their proper chemical
relations. To my surprise I have found in the tuberous root of
yams a slimy matter that is precipitated from its solution by dilute
acetic acid. Further investigation has demonstrated beyond any
doubt, that this slime belongs to the class of mucins, and as this
remarkable occurrence is the first exception to a generally adopted
rule, I will describe in the following lines my observations in detail.

1. Short Description of Japanese Yams.

There are two kinds of yams growing in this country, the
one is called “yamano-imo” (Dioscorea japonica, Humb.) while the
other is called ‘‘jimenjo”’ (Dioscorea batatas, Decaigne.) Both of
them are found in a wild state and are very often cultivated.
“ Naga-imo,” ‘‘tsukune-imo”’ and ‘‘ichinen-imo’’ are indeed the
names given to the three different cultivated forms of ‘‘jinenjo.”’
All of these forms produce the large tuberous roots which are
used as food.

98 ON THE OCCURRENCE OF MUCIN IN PLANTS.

Composition of the yam.")

Water 80.74
In roo parts of dry matter,
Crude protein 11.74
Fat 0.84
Fibre 4.36
Ash (free from Co,) 3.60
Starch 22, AZ
Other non-nitrogenous substances. 57-33
Total nitrogen 1.879
Nitrogen in amides, etc. 0.675

2. Preparation of the Slime of Yams.

The slimy matter forms a thick turbid liquid and can
be easily freed from starch granules and other substances by
simply filtering. The filtered liquid shows a neutral reaction,
and is precipitated by the addition of acids, but the pre-
cipitation is prevented to a certain extent by the presence of
common salt. Several methods are proposed for the isolation of
mucin.

Landwehr” prepared the mucin of the bile by precipitating
with acetic acid, washing, dissolving in 1%. solution of soda,
and precipitating again. Obolenski>) recommended a method of
purifying the mucin of the submaxillary glands as follows: The
glands are soaked in water over a night and filtered. The filtrate
is precipitated with acetic acid, the precipitate washed first
with water and a little acetic acid, then with hot alcohol and
finally dried. But Hammarsten*) recommended a new method
which is well adapted to separate mucin from substances belong-
ing to the group of nucleoalbumins. The mucin of the sub-
maxillary glands is soluble in dilute hydrochloric acid (o.1—
0.2%) and is precipitated unchanged by adding 3 or 4 times
its volume of water to the solution, while the nucleoalbumin
dissolves in dilute hydrochloric acid together with the mucin,

1) The analysis was made in the agricultural chemical laboratory of the Im-
perial College of Agriculture in Tokyo. See Bull. Vol. I.

2) Z. physiol. Chem. Bd. V, 371.

3) Pflig. Arch. 4.
4) Z. physiol. Chem. Bd. XII.

ON THE OCCURRENCE OF MUCIN IN PLANTS. 99

but is not precipitated by adding water. As the precipitate
obtained by the addition of acetic acid to the extract of yam
root was insoluble in dilute hydrochloric acid and with difficulty
soluble in very dilute alkali (as 1% solution of caustic soda), I
worked chiefly according to the method of Obolenskt.

The roots were cut into small slices, well crushed and
extracted with about three times the volume of water under
frequent stirring; the thick liquid thus obtained was filtered,
and the filtrate precipitated with a very dilute solution of acetic
acid gradually added. The addition of a large amount of acetic
acid at once, has to be avoided, otherwise the liquid will become
so thick, as to make filtering impossible. The precipitate was
washed with dilute acetic acid, and then with dilute hydrochloric
acid for the purpose of washing away a trace of another proteid
soluble in the latter acid; the precipitate was finally washed
with water, with alcohol and ether, and at last again with
absolute alcohol. Upon drying, the purified substance formed
a hard yellowish mass.

3. Reactions and Composition.

The substance obtained has the following chief reactions:

It dissolves in caustic alkali of about 2%, but with difficulty,
and is soluble in strong mineral acids as well as strong acetic
acid.

It is not digested by artificial gastric juice, but easily by an
alkaline solution of trypsin.

When concentrated sulphuric acid is added to its solution
in acetic acid, it yields a fine violet coloration.

It gives the xanthoproteic and biuret reactions.

By boiling with Millon’s reagent, it forms a red precipitate.

Tannin precipitates its solution.

Double iodide of potassium and mercury yields a turbidity.

By boiling for some time with 5% sulphuric acid, it yields
not only peptone, but also a substance which reduces Fehling
solution.

The elementary analysis was made with the purified sub-
stance, dried at r10°C. to constant weight, and yielded as
average of three experiments the following numbers:—

suelo) ON THE OCCURRENCE OF MUCIN IN PLANTS.

C 52,82
H 7:53
N 14,20
O-+S (calculated) 25,05
Ashes 0,41.

The determination of N was made after the method of
Kjeldahl. The substance contains sulphur. By boiling it with
caustic potash of 5%, the liquid became yellowish, and when
hydrochloric acid was added in excess, sulphuretted hydrogen
gas was evolved, and the paper moistened with lead acetate
turned black owing to the formation of lead sulphide.

The composition of this slime is approximately the same
as that of the bile (see the following table). Identity is of
course doubtful, as there are many isomeric, especially stereo-
isomeric mucins, possible.

C lel INI SO) Ashes

Mucin of tendon?) 48.30 6.44 11.75 0.81 —— --- 5
Mucin of submaxillary glands?) 48.84 6.80 12.22 0.84 —— ——
Mucin of bile?) 53-09 7.60 13.80 1.10 24.41 ——
i o_o"
Mucinous substance of yam roots 52.82 7.53 14.20 25.04 0,41

Hammarsten declares: ‘‘Die Fahigkeit, beim Sieden mit
verdtinnten Sduren eine reducierende Substanz zu geben, die
physicalische zihe Beschaffenheit und das Verhalten zu Es-
sigsdure sind die drei wichtigsten Eigenschaften, welche die
Mucinstoffe in qualitativer Hinsicht von dem Eiweissstoffen
unterscheiden.’”’ Our slime shows all the essential charac-
teristics of the animal mucin, and differs only in some sub-
ordinate points; it is with more difficulty soluble in dilute
alkalies and yields a turbidity with potassium ferrocyanide.
Our substance is doubtless a kind of mucin, and as this is
the first time that such a compound has been found in the
vegetable kingdom, it is certainly of physiological interest.
The quantily determined as accurately as possible, amounts
to 8% of the tuber dried at 100°C.

1) Loebisch, Ztschr. physiol. Chem. Bd. X, S. 61.
2) Hammarsten, Ztschr. physiol. Chem. Bd. XII,
3) Landwher, Ztschr. physiol. Chem. Bd. V, S. 371.

Mannane as a Reserve Material
in the Seeds of Diospyros Kaki, L.

BY

J. Ishii, Nogakushi.

The fruit of Diospbyros Kaki, L. (date-plum) is consumed
in large quantities by the people of this country on account of its
richness in saccharine matters. There are many varieties of
the fruit, ranging in size from a small hen’s egg to a large
apple. In color’) of the epidermis they vary from light orange
yellow to deep orange red. My preliminary experiments proved
it probable that a wine of good quality might be prepared
from the fruit. It may be also mentioned here that this fruit
contains, when unripe, a considerable amount of a kind of tan-
nin which disappears entirely during the ripening of the fruit.
My investigations of the flesh of the fruit have shown that
there is a great amount of dextrose and laevulose, but neither
mannose nor galactose present. It 1s, therefore, surprising that
the sceds of the fruit contain no trace of starch, but a soft
white mass as a reserve material, which could be easily con-
verted into a sugar by boiling with sulphuric acid of 5%
for one hour. After removal of the sulphuric acid with barium
carbonate, the filtrate was evaporated, when a red-colored sub-
stance was gradually deposited; this was filtered off and the
solution after being decolorized with animal charcoal was
further concentrated. I obtained a sweet syrup which yielded,
with acetate of phenylhydrazin in the cold, a considerable
amount of crystalline precipitate, which upon recrystallization
formed white tabular rhombic crystals, melting at 195°C (man-
nose-phenylhydrazon). By mixing a certain quantity of acetate
of phenylhydrazin with the aqueous solution of the crystals
and heating the mixture, there were formed gradually yellow

1) This coloring matter is turned blue by treatment with concentrated sul-
phuric acid.

102 MANNANE AS A RESERVE MATERIAL.

needles soluble with difficulty in hot alcohol and melting at
205°C. evidently phenylglucosazon. ‘Therefore, there can be
hardly any doubt that our sugar is mannose, and the white
substance in the seeds a polyanhydride of mannose called man-
nane. It is physiologically highly interesting to see that the
seed stores up here in the form of an anhydride a sugar that
is different from the sugars contained in the flesh of the fruit.

Mannane as an Article of Human Food-

BY

C. Tsuji, Nogakushi.

Since the discovery of mannose by Reiss by treating the
so-called cellulose of the nut of Phytelephas with sulphuric acid,
and since its closer description by E. Fischer who obtained it
as a product of the oxidation of mannite, there have been found
in many plants slime-like and cellulose-like polyanhydrides that
yield by hydrolysis mannose. They have been distinguished as
mannane, para-mannane and manno-cellulose.’»

Thus far, there is not known, however, any article of human
food whose nutritive value is due to a polyanhydride of mannose,
while polyanhydrides of glucose (starch, glycogen, maltose) play
an important role, especially starch.” But there are sold in
this country as an article of food gelatinous colorless tablets,
apparently consisting of starch paste. called namakonniaku that
are largely consumed by the people. These tablets, however,
do not give a blue coloration with iodine, and do not, there-
fore, consist of starch. My investigation has proved that this
substance is a polyanhydride of mannose. In the following
lines, I will describe the root from which the konakonmiaku is
made, and the experiments which elucidate the nature of the
product.

This tuberous root-stock is derived from Amorphophallus
Rivieri Durien, var, Konjac Engl., a plant belonging to the
Aroideae and largely cultivated in the central part of this
country. The root resembles in form the taro, has a white
spongy flesh of a sharp taste and a brown epidermis. The root-
stocks vary in size from that of a potatoe to that of a squash
and reach sometimes the weight of several kilos.?)

1) Compare E. Schulze, Z. Physiol. Chem. 16, 386.

2) The refuse of the stone-nut, used in manufacture, is utilized now in some
places as a food for animals with good effect.

3) The art of preparing a suitable food from this root-stock and of removing
the sharp taste was introduced into this country about a thousand years ago by
Chinese.

104 MANNANE AS AN ARTICLE OF HUMAN FOOD.

There are prepared a powder and a gelatinous mass from
the root-stock. To prepare the powder, the root-stock is sliced
into thin pieces after all the skin is removed. These slices are
hung up to dry, and after several weeks are ground in a mortar,
and sifted.

To prepare namakonniaku, as the gelatinous mass is called,
the powder or the root-stock is well boiled with water, then
ground into a pasty mass, forced through a sieve, and transferred
into a large wooden tub, mixed with an equal quantity of slaked
lime and double the amount of water, and kneaded with the feet.
After the mixture has become homogeneous, it is boiled with
lime water until it forms a gelatinous mass.”

Chemical Investigation.

The powder of konntaku above mentioned served principally
for my experiment. It was boiled with 3% solution of sul-
phuric acid for several hours. On filtering a yellowish solution
was obtained which was neutralized with barium carbonate,
decolorised with animal charcoal, filtered, and evaporated to
a syrup, from which no crystals could be obtained. It was
soluble in cold water and dilute alcohol, had a strong power of
reducing Fehling’s solution, had a very sweet taste and turned
the plane of polarization to the right. This syrup consists
evidently, to a great extent, of mannose, for even a very small
quantity of it at once gave in the usual way treated in the cold
with solution of acetate of phenylhydrazin, a colorless crystalline
precipitate soluble in hot water and hot alcohol, and thus very
easily purified. The melting point of this purified precipitate
was found to be 195—200C?.

There can be no doubt that this was mannose-phenyl-
hydrazon, as this substance was easily transformed by further
treatment with phenylhydrazin into phenyl-glucosazon melt-

1) The workmen have to avoid inhaling the fine powder by covering the nose
with a cloth, as the powder irritates the throat.

2) This mass is made to freeze in winter, whereby its aspect changes; this
product is preferred and is called “‘ kovikonniaku.”

From the root-stocks, a kind of paste is made which is principally used to in-
crease the lustre of clothes and which is far superior to starch paste, and cheaper
than the latter. Paper treated with this paste of kunniaku is used instead of oiled
paper for our umbrellas and water-proof clothes.

MANNANE AS AN ARTICLE OF HUMAN FOOD. 105

ing at 205°C. The characteristic mannose-oxime was also ob-
tained by slow evaporation of a portion of the syrup with a mix-
ture of hydrochlorate of hydroxylamine with sodium carbonate
and purified by recrystallization from absolute alcohol. A por-
tion of the sugar-syrup was also repeatedly oxidized with nitric
acid to see whether mucic acid could be obtained, but none was
found; therefore no galactan was present in the konniaku root-
stock. Also, as no pentose reaction with hydrochloric acid
and phloroglucin could be observed, there could not be present
in the konniaku-root xylan or araban to any considerable extent.

If all the sugar obtained was mannose, as is very probable,
then the konniaku powder mentioned yielded 55.86% of mannose.

This konniakw root is evidently very well suited for the
preparation of the polyanhydride of mannose called mannane
in a pure state. As this mannane is used as food, it must
evidently be digested by the enzymes in the intestines and
transformed into mannose or a dimannose corresponding to
the maltose made from starch, but my attempts to convert the |
mannane of konniaku by diastase made from malt into a sugar,
were not successful. The more interesting then is it that the
human intestines can digest mannane.

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On the Scale Insect of Mulberry Trees.
(DiaSPIS PATELLIFORMIS N. SP.)
BY
C. Sasaki, Rigakuhakushi,
Professor of Entomology, Agricultural College, Impertal University,
Tokyo, Fapan.

WitTH Prates I.—II.
I). Description of the Mature Insects.

Male.—The body is orange red; legs, antenna, veins of
wings, balancers and anal filament yellow; head comparatively
small, slightly pointed in front, where are two long and slender
afitenne (Fig. 1; Fig.1, a; Pk 1). The antennz (Fig. 1, d) are
composed of ten segments, of which the first is short, large, and
stout; the second shorter and smaller than the first; the re-
maining eight segments are cylindrical and nearly equal in
size and form, while the terminal tenth segment has a pointed
end. Its length is 0,7 mm.

Eyes (ocelli) are globular, prominent, and four in number,
two on the dorsal and two on the ventral side of the head (Fig.
1,bandc; Pl. I.). The dorsal pair of eyes, which lie near the
lateral sides of the head, are reddish ochre brown and are
surrounded by a dark brownish ring at the base. The ventral
pair, which are similar in size, form, and color, to the dorsal
pair, lie a little removed from the lateral margin of the head.

The thorax is ovate oblong, its anterior edge is nearly
straight. The three segments—pro-, meso-, and meta-thorax—
are more or less marked off from one another. The pro-thorax
which is small, nearly quadrangular in form on its dorsal side,
is much longer and broader on its ventral side, and its tergal
face is marked with two dark orange inclined lines.

The meso-thorax which is the largest of the thoracal seg-
ments is somewhat angular in front, and rounded at the posterior
end. The tergal side of the same segment is vaulted, and its
scutum is marked with a dark broad reddish orange transverse

~-7 7 0?

108 ON THE SCALE INSECT OF MULBERRY TREES.

band on the anterior edge, the remaining portion with three lon-
gitudinal parallel lines. The postscutellum is small, nearly
triangular in form, and there lies, between the scutum and post-
scutellum, a broad transverse band of dark reddish orange color.
The meta-thorax which is the smallest of the thoracal segments,
is wide, and closely jointed on its entire breadth with a first
abdominal segment, so that it looks like one of the abdominal
segments.

The fore wings, thin, membranous and transparent, are long
oval, narrowed at their base. They are provided with two
nervures—subcostal and cubital. The former is longer and
stronger, passing parallel with the costal margin, disappears
gradually towards the external margin of the wing. The
cubital nervure is shorter and finer than the former, arises near
the base of the subcostal nervure, and running obliquely ter-
minates about midway of the posterior margin of the wing.

The balancer (Fig. 1, f. Pl. I) is composed of two parts
nearly equal in length, of which the distal has a form of bristle
with a free end slightly curved.

The legs (Fig. 1, e. Pl. I) are all of a moderate size, and
almost similar in length and structure. The coxal joint is oval
and stout, the trochanter which is oblong in form, is closely
attached to the femur, so that these two seem to form a single
spindle-shaped segment. The tibia which is somewhat more
slender than the femur, is beset with a few short hairs. The
tarsus is formed of only a single long piece pointed at the free
end, which bears a pointed claw. The tarsus is also provided
with short hairs as on the tibia. At the insertion of the claw on
the tarsus, are three long club-shaped hairs,

The abdomen is nearly equal in length to the thorax, but
is gradually narrowed towards its free end. It is jointed with
the thorax by its entire breadth, and is composed of nine seg-
ments. The terminal segment, is provided with two long bristle-
like appendages lying close side by side, so as to form a single
long filament measuring 0,25 mm. in length.

The length and breadth of the body are on an average 0,80
mm. and 0,25 mm. respectively. The expansion of wings is
2,0 mm. and the caudal filament is 0,25 mm. in length (Fig. 15
dean, ele li)

ON THE SCALE INSECT OF MULBERRY TREES. 109g

Female.—The body is flattened nearly oval, light yellow in
color, and covered sparsely with either club-shaped or simple
hairs. It is composed of nine segments and on the lateral
margins of the segments from the 3rd to the goth, there are beset
a number of long simple or 2-3 divided spines ending with an
extremely fine slender filament. The dorsal surface of the body
is marked by several depressions or wrinkles, and its anterior
portion is broader and rounded, while the posterior is abruptly
narrowed towards the free end. There is generally no demar-
kation between the head, thorax and abdomen, and the entire sur-
face of the body is marked with very fine wavy striations lying
very closely to each other. The lateral sides of the body are
marked symmetrically with blunt processes of unequal size, which
however indicate the segments of the body. The segment-lines
of the body are rather hard to perceive, but on the posterior
portion they become conspicuous. Besides these processes, the
segments of the body are to be recognized by rows of oval figures,
which lie along the boundary line of a few posterior segments.
Judging from the boundary lines marked with figures, as well as
the lateral processes, we may say that there are nine segments,
which compose the body (Figs. 2, and 3, Pl. I).

Pygidium.—This is composed of the two last segments (8th
and gth), and is much more chitinous and hard than the remain-
ing segments (Figs. 4, and 5, Pl. I.). The dorsal surface of the
pygidium is deep orange yellow, and of an almost triangular
form. Its surface is uniformly marked out with longitudinal
parallel fine striations, and a little above the middle portion
of this plate lies a single round opening (anus), while all over
the surface are several elongated marks. On either side of the
dorsal side of the pygidium, that is, on the boundary line between
the 7th and 8th, and the 8th and oth segments, there lies a row
ot oval figures mentioned above, which are more or less irregu-
larly arranged one after another. These rows of oval figures are
not limited to this region, but two imperfect rows of them may
also be found on each of the boundary lines (dorsal) of the 5th
and 6th, and the 6th and 7th segments. These oval figures
secrete delicate transparent band-like materials which are used
as the constituents of the scales (Fig. 4, Pi. I.). When highly
magnified, these lines are more or less conspicuous by a darker

II0 ON THE SCALE INSECT OF MULBERRY TREES.

color than the remaining portion of the body. The ventral side
of the pygidium is nearly similar in color, form, and nature to
the dorsal. The lateral margins of the pygidium are irregularly
dentated, and bear a few spines bifurcated, or divided into three
short branches ending with a fine slender filament, and also a
few short simple hairs. At the free narrow posterior end of the
pygidium, there lie two short and wide processes arranged
very close side by side. At the portion nearly anterior to one
third of the ventral side of the pygidium, there lies a small
genital opening. On the three sides of the genital opening, there
can be seen five groups of very small polygonal or circular areas,
of which one is on the anti-median portion of the opening, and
the remaining four (two on each side) on the lateral portion of
the same. Each of the groups is more or less oval and elongated,
and they lie close to each other end to end. These polygonal or
circular areas are marked on their surface with a number of
minute dots. They are considered as secretory pores by Tozetti,
and Franceschini‘ in the species Diaspis pentagonia, Targ., but
I think in my specimen, they are not secretory pores, being
simply a sort of markings (Fig. 5, Pl. I.). The antenne (Fig. 6,
Pl. I.) are two in number, and lie close at their base on the
ventral side of the first segment near its free anterior edge.. They
are formed of a single stout broad piece, having three processes
of variable size, at the base of which is beset a single long bristle
(Fig. 6,° Pl. -1,).

The mouth parts (Fig. 7, PL. I) are modified into four long
filaments (mandibles and maxilla) which are inserted on a
small process at the ventral side of the first segment of the body.
They arise from one and the same point, and all of them are
closely applied to each other, so as to form a single long brown-
ish thread, whose length (1,2 mm.) is a little less than that of
the body. Thc insect remains attached to the bark by penetrat-
ing the thread deep into the bark of the mulberry tree in order
to suck the nourishment.

Anterior to the insertion of the thread, there lies a nearly
triangular and somewhat concave space, which is protected on
all sides by a rectangular chitinous ring which lies just beneath
the skin. This concave space seems, however, to have a func-
{ion similar to the sucking discs on the arms of the cuttle-fish,

ON THE SCALE INSECT OF MULBERRY TREES. Gi

in order to attach the insect-body tightly to the surface of the
bark. On the front corners of the rectangular chitinous frame-
work, lies a wide process of the same chitinous nature, while
posterior to this framework is inserted the root of the four
modified filamentous mouth parts. Interiorly from this root are
projected two pairs of chitinous rods, of which one seems a part
of modified mandibles and the other, that of modified maxilla.

The three sides (anterior and lateral) of this concave space
are somewhat swollen, and marked off from the rest of the body
by a horseshoe-shaped line.

There are two pairs of spiracles (Fig. 8, a, PL. 1) one on
the ventral side of the first segment, and the other on the
same side of the third segment of the body. The former
pair open on either side of a process, on which the modified
mouth-parts are inserted. They are simple oval openings
measuring 0,01 mm. in longer diameter. Round this opening,
except on its posterior sides, there lies a hemispherical area
marked with a number of round spaces having a few fine
‘ perforations (Fig. 8, PL. I.). These perforations seem to
secrete sticky filaments, which lie always in a clustre over the
spiracle in the form of a white mass.

The second pair of spiracles are simple, small, and nearly
circular openings, and they lie apart from each other on the
ventral side of the third segment. They are smaller than those
on the first segment, and measured 0,0073 mm. in diameter (Fig.
Sa. PL} J.),

The skin is pretty thick, transparent and strong, and there
may be found a large number of very fine irregular transverse
striations on the entire surface of the body except the pygidium,
which is, on the contrary, marked with longitudinal striations.
In addition to the striations, the body is covered with fine short
hairs, which grow somewhat thicker near the periphery of the
body.

The average length and breadh of the female insects are 1,3
mm. and 1,0 mm. respectively.

The scale which covers the female insect is more or less
round, oval, and flat, bearing a slightly projecting apex nearly at
the anterior third of the scale, and thus it takes the form of a
patella, whence the specific name “ patélliformis”’ is given. It

I12 ON THE SCALE INSECT OF MULBERRY TREES.

is pretty thick and tenaceous, but is gradually thinned out
towards the peripheries. The surface of the scale is of a
brownish grey color, and bears on its apex always two oval
areas of orange yellow. These two areas are really two
moulted skins, and the smaller lies over a portion of the larger.
The former is somewhat lighter in color than the latter (Fig. 9
PL. I.). The average diameter of ten large scales is 2 mm.

II.) Manner of Copulation.

From June to the beginning of November, both sexes of the
scale insects appear, the females remaining tightly attached
to the bark of the mulberry tree, protected by a patella-like thick
scale, which may be formed by the secretion from their
bodies. The males, on the contrary, being active and provided
with a pair of developed wings, can either walk or fly. When
we watch carefully the scales on the bark at this season, we find
that many winged males approach the scales on their wings.
When the male finds the scale, the former rests quite close to
the latter, and then it bends its long bristle-like genital append-
ages (caudal filaments) towards the margin of the scale, and
projects them inwards under the scale, and copulates. Although
Iam unable to state exactly how the male conveys the male
elements to the female, yet that such action of the male against
the scale, that is, the penetration of the male genital appendages
under the scale is a process of copulation, is proved by the
examination of the contents of the female insects, in which there
may be found a large number of actively moving spermatozoa
which are received from the male. The males generally die at
the end of a few days after copulation, while the females pass
the winter under a scale. Thus our scale insect increases by
sexual reproduction which can not be observed in Diaspis penta-
gonia, as Mr. Contagne has mentioned.

III.) Metamorphosis of the Scale Insects.

About March, the body of the female insects is more or less
swollen by a large number of developed eggs within the body.
The ovarian eggs are numerous, but the large eggs contained,

ON THE SCALE INSECT OF MULBERRY TREES, Il3

wd.

vary from a hundred to a hundred and fifty in number. The
larger eggs are oblong oval, transparent, and measure 0,05 mm.
and 0,025 mm. in the longer and shorter axis respectively. The
light greenish contents of the eggs are easily seen through the
transparent egg-shell. In May and June as well as August and
September, the female insects lay eggs by projecting a temporary
fleshy process from a genital opening on the ventral surface of
the pygidium (Fig. ro, Pl. I.). All the eggs laid, lie usually in
groups beneath the body of the mother insect under a scale.
Even after the mother insects have finished depositing their
eggs, they remain alive for some time. A single female deposits
usually about a hundred eggs much larger than the ovarian
eggs which were found in the previous spring, but they still
retain an oblong and oval form, and measure now 0,247 mm.
and 0,114 mm. in the longer and shorter axis respectively (Fig.
11, Pl. I.). The newly laid eggs are always pale yellow, but
later as the embryo is fully developed within the egg, the latter
changes from light to deep orange yellow. The larve are
hatched some days after being deposited, and they crawl about
from fissures or cracks formed at one or more places along the
marginal edge of the scale. At the moment the young larve are
hatched, the transparent thin egg-shell breaks always in one
and the same way, that is, from one pole of the egg extends a
breaking line as far as the median portion on two sides of the
egg-shell. The newly hatched larve (Fig. 12; 12, a. Pl. I.) are
oval flat, light orange red in color, the length and breadth of
the body 0,266 mm. and 0,193 mm. respectively. The body is
covered with a few fine short hairs over its entire surface, and
its cuticula is marked all over with very fine wavy lines which
are closely arranged side by side (Fig. 1a, c. Pl. I.). The
three regions of the body are not distinct, and the latter is
composed of nine unequal segments, of which the anterior first
segment is much larger than the last, that is the ninth segment,
while both are hemispherical in form. The remaining seven
segments are short, but their breadth exceeds that of the two
extreme segments. The anterior first segment is marked on
either side of the dorsal surface with a longitudinal slight de-
pression. Onthe front of the same surface there lie two small
black simple eyes lying wide apart from each other, and further

I14 ON THE SCALE INSECT OF MULBERRY TREES.

on the front edge are beset two small bristles. Anteriorly on
the ventral surface of the first segment are provided two long
antenne. Each of them is composed of five segments, of which
a single terminal one is nearly of the same length as that
of the four remaining segments taken together. The antennal
segments are provided with a few hairs, and the terminal
segment with a single long hair.

The mouth parts, which are similarly modified as those of the
mother insects, lie also on the ventral side of the first segment,
and the position where they are inserted is marked off with a
slight elevation. <A filament formed of modified mouth parts lies
beneath the cuticula of both thoracal and adbominal segments.
It runs straight to the middle portion of the adbomen, and the
running up to the first thoracal segment, turns again downwards
so as to form a sort of long ovalloop. The last, that is the ninth
segment, is provided with two stout chitinous processes at its
free end, between which are inserted two long slender bristles.

The three pairs of legs on the thoracal segment are of the
same size and of similar structure. They are short and stout,
the coxa and trochanter are distinct, the femur is nearly oval
and the largest of all the segments, the tibia and tarsus are
amalgamated into a single long segment, but the former, much
reduced in size, is marked off from the tarsus by a fine trans-
verse line. The tarsus is long, slightly widened at the free end
bearing a simple claw, and the former has a slight notch just
behind its free end (Fig. 12, b. Pl. I.).

The larve of this stage are very active, and they crawl
about in every direction on stems or branches particularly in
cracks. After remaining in this condition 3-4 days without tak-
ing nourishment, they undergo a first moult.

When the larve has ended its first moult, it grows to a size
of 0,38 mm. and 0,18 mm..in length and breadth respectively,
and its color becomes paler than in the previous stage. Further
on, it attains 1,0 mm. in length, and undergoing a second moult,
becomes either a male or female pupa.

In this larval stage (Fig. 13,14. Pl. I1.), a filamentous mouth
part (rostral setae) becomes free, and the larva, projecting it deep
into the bark of mulberry tree to imbibe the sap, remains attach-
ed to the same spot without changing any longer its position.

ON THE SCALE INSECT OF MULDERRY TREES. 115

The extended rostral setae are comparatively long, measuring
0,47 mm. in length, and are yellow incolor. The body is covered
with sparse fine hairs, and its form as well as the antenne,
legs, the two processes and bristles on the last segment, is similar
to those of the previous stage, the only difference being the in-
creased size. The chief characteristic in this stage is the
presence of two small openings (secretory pores) dorsally on the
space between the two simple eyes (Fig. 13. Pl. II.). These
openings lie at the end of a short chitinous tube embedded with-
in the body, and from each of the openings, is spun out a sticky
continuous silky thread which covers loosely the hinder portion
of the larval body. When the threads are thus spun out, they
dry up by exposure to the air, and the mucous envelope (?) of the
threads is peeled off in a form of small hemispherical pieces,
which present the appearance of white dust on the posterior
portion of the larve (Fig. 15, 15, a. Pl. II.). On either side
of the mouth region, that is, at the base of the secretory pores,
there may be seen through the skin a concentrically coiled
thread, which seems to be of the same nature as silky thread
spun out of the secretory pores.

The spiracles (Fig. 14. sp. Pl. II.) are silaple and small open-
ings. One pair of them opens ventrally on the first and third
segments of the body. In this stage, it seems there is still no
distinction between male and female larve.

Some days after the larve have attached themselves to the
bark,they moult again and change to either male or female pupe.
At the time of moulting, the skin of the larve splits up lengthwise
on the ventral side, and there comes out a pupa, which remains
still for some time beneath the moulted skin, while later the body
of the pupa protrudes posteriorly from the moulted skin.

The male pupa is oval, somewhat depressed, sac-like in
form, measuring 0,5 mm. and 0,3 mm. in length and breadth,
and has a light greenish yellow color (Fig. 16, 16, a. Pl. II.).
It has no limbs or even the rudiments of wings; but the pos-
terior portion of the body is marked with some transverse lines
showing a number of segments. The three regions of the body
are not distinct. The two pairs of dark reddish brown ru-
dimentary eyes lie near the front edge of the body, and still
anteriorly on the ventral surface of the same, there is projected

116 ON THE SCALE INSECT OF MULBERRY TREES.

a long slender rostrum, with which the pupa imbibes the sap.
If we now closely examine the body, there may be found a
number of small openings on the dorsal and ventral sides of each
segment from the znd to the oth (Fig. 16, b. Pl. II.). From
these openings, are secreted very fine white filaments, which ac:
cumulate on both dorsal and ventral sides of the pupa in the
form of a sac, opening at the free edge, just behind the moulted
larval skin. This is the beginning of a cocoon, later it grows
longer posteriorly till it becomes a perfect cocoon,

The perfectly formed cocoons (Fig. 17. Pl. II.) are snowy
white, long and oval in form measuring 1,4—1,6 mm. in length
and 0,4—0,5 mm. in breadth. The surface of the cocoons is
usually marked with a few longitudinal ridges. They are at-
tached to the bark of mulberry trees by their smaller end, which
carries always dorsally a moulted larval skin colored greyish
yellow, while the broader end is more or less elevated, and re-
mains open free.

Some days after the pupa has been imprisoned within the
cocoon, the latter bears partly an orange color, and the pupa
contained within, growing now 0,67 mm. in length, is of a some-
what dark orange color. At this period of development, the body
of the pupa is more or less depressed, and colored orange red
(ig. 18. Pl. II.). The three regions of the body are more or
less distinct, the segments of the thorax and abdomen come
into view, and the eyes are developed into a globular form of a
dark brownish red color. The antenne, wings, three pairs of
legs, and a caudal appendage grow on the body in a sort of
cylindrical sacs. The antennze and wing processes lie closely
attached to the lateral sides of the body. Of the three pairs of
legs, the first pair is directed forwards, the remaining two pairs
toward the posterior end, and a single caudal appendage lies at.
the end of the abdomen in the form of a blunt process (Figs. 19;
20. Pl. II.). When the pupa is so far developed, it loses its
filamentous rostrum, and no longer takes sap from the bark,

The male insects are hatched twice a year, viz. in June
and October, in most cases. They come out generally through
the free open end of the cocoon from their posterior end;
but sometimes they get rid of the anterior fixed end of the
same.

ON THE SCALE INSECT OF MULBERRY TREES. Ii7

After the winged insects have appeared, they remain gener-
ally near the cocoon a little while, and then crawl about the
female scale, or fly in the air in order to search for the scales.

The cocoons of the male insects are generally found in
groups on either stems or branches, and are particularly nu-
merous near the base of the stem, where it is protected from
the sunshine by overlying branches and leaves, and thus the
cocoons give the latter a snowy white appearance in patches of
variable size or at all the sides like girdles round the stems or
branches (Fig. 21. PL. II.)-

The groups of the cocoon are loosely covered with white
continuous filaments bearing scattered white dust: these fila-
ments are those secreted by the larve, and the dust is hemi-
spherical in form. The continuous filaments are enveloped with
a sort of mucous layer more or less sticky in nature, and as this
layer dries up in the open air, it is thus peeled off in pieces
having the appearance of small snowy white dust as mentioned
before.

Female pupa.—When the mature larva fixes itself tightly
with a filamentous rostrum on the bark, a scale begins to form
on its posterior portion. This shows the presence of a newly
formed pupa beneath the old larval skin as well as the fragment-
ary scale. The pupa (Fig. 22. PL. II.) is oval depressed,
measures about 0,5-0,4 mm. in length and breadth respectively.
The body is composed of the same number of segments as the
mature female insect, and is also very similar in general aspects.
The head segment bears two rudimentary antenne, two dark
brownish red eye spots, and a filamentous rostrum. The pupa
with its rostrum imbibes also the sap. The abdominal segments
aré more or less conspicuous, and near the lateral margin and
close to the boundary line between the seventh and eighth
segments, as well as the free margin of the chitinous shield
(pygidium) (Figs. 23; 24. PL. II.), are marked with some
small oval openings at both dorsal and ventral surfaces. These
openings secrete, without doubt, an extremely fine filament used
as a material to form the cocoon. Further dorsally on the
pygidium there opens an anus in the form of a small round
opening. The free pointed end of the pygidium is provided with
a pair of broad chitinous processes, and each of the lateral

118 ON THE SCALE INSECT OF MULBERRY TREES.

margins of the same is beset with four pointed strong processes
arranged at regular intervals. When a nearly circular depressed
cocoon (scale) attains about I mm. in diameter, the pupa be-
neath it undergoes the last or third moult. The moulted skin
which is of a dull brownish red color, becomes also part of the
scale, thus the brownish red mark on a fully formed scale is
nothing else than the moulted-skin of a pupa (Fig: 9. b. PL. I.).

At the moment of moulting, the pupa-skin is split up lon-
gitudinally on its ventral surface, and a female insect may
come forth. Remaining in the same position, that is, exactly
beneath the moult-skin, it attaches itself firmly to the bark by
penetrating it with its long filamentous rostral sete. The newly
hatched female scale insects secrete also fine threads from the
pores, which open on the posterior region of their bodies, and
these secretions accumulate at the periphery of the already
formed scale, thus increasing its dimensions. In this way the
scales attain their normal size.

The scales (Fig, 21. PL. II.) are formed either in groups or
are scattered sparsely over the bark of twigs or branches, and
most of them, lying beneath the epidermis of the bark, present
a dull greyish yellow aspect, which makes it very difficult for
us to observe (especially when they lie very sparsely), owing
to their color being similar to that of the bark.

As I have mentioned before, our scale insect breeds only
twice a year; instead of three times, like the species Diaspis
pentagonia Targ. observed by Mr. Y. Contagne."

IV.) Determination of the Species of the Scale Insect.

The want of sufficient literature on the scale insect makes
it very difficult to determine its exact species. But still we
may say the scale insect of the mulberry trees unquestionably
belongs to the genus Diaspis whose characteristics Mr. Albert
C. F. Morgan’? has described in his work ‘‘ Observations on
Coccidae.’’ Its specific characters agree to a certain extent
with those of the Diaspis. pentagonia, Targ., which Messrs
Targioni-Tozetti and Franceschini,’ and also Mr. G. Coutagne’
have very accurately described; but still some differences may
exist between Diaspis pentagonia, Targ, and our Japanese scale

ON THE SCALE INSECT OF MULBERRY TREES. IIg

insect, in all three different stages,—larva, pupa and imago.
These differences, I think, afford sufficient grounds to justify me
in giving a new specific name, ‘‘patelliformis,”
insect.

In the following lines I will mention all the characteristics
of male, female, and larva of our scale insect, which differ from
those of Diaspis pentagonia, Targ. described by the above men-
tioned authors.

to our scale

I. Male.

a. Antenne are composed of ten segments, of which the
terminal one is rather long and slender, and somewhat pointed
at the free end.

b. Of the three segments of the thorax, the pro-thorax
is nearly quadrangular, the meta-thorax is short and wide, and
is closely jointed along its entire breadth to the first abdominal
segment, and is not triangular in form.

c. The abdomen is long and conical, not elliptical.

d. The balancer is composed of two portions, the basal
half club-shaped, while the distal half is reduced into the form
of bristles with a hook-like end.

e. The tibia of the leg is long and cylindrical, not trian-
gular in form. At the insertion of a claw on the tarsus, there
are beset three long hairs of nearly equal length, bearing a small
round head at their free end.

II. Female.

a. The body is composed of nine segments which are
indicated by sutures (segment-lines) as well as by the rows of
secretory pores.

b. The antenne are formed of single broad segments
with their free end divided into three pointed branches, and near
the base of the antennz there is a single long bristle.

c. One or two rows of secretory pores lie dorsally along
the suture of the 5th-6th, 6th-7th, 7th-8th, and 8th-gth seg-
ments.

d. The bristles on the pygidium are either simple or
divided, having on each free end a very Jong fine filament.

120 ON THE SCALE INSECT OF MULBERRY TREES,

III. Larva in the first stage.

a. The body is composed of nine segments.

b. On the circumference of the body there are no
depressions.

c. The antenne are composed of five segments.

d. No particular depressions at the insertion of the
antenne. :
e. The rostral setae lie beneath the skin.

IV. Larva in the second stage.

f. There are two spinnrets dorsally on the first segment
of the body.

g. Dorsally and ventrally along the sutures of the
posterior segments, there lie some secretory pores.

h. The rostral setae become free and unrolled.

In addition to these differences, we also notice that our
scale insect copulates, and is not parthenogenetically re-
produced, and the generation. is effected only twice a year,
not thrice as in Diaspis pentagonia, Targ.

V). Damage done by the Scale Insects,
and Preventive Measures.

The scale insect is widely distributed over nearly all the
provinces of Japan, where the mulberry tree is cultivated. It is
not only parasitic on the mulberry trees, but may also be
found in many other plantations.—such as Brousonetia papy-
rifera, Prunus pseudo-cerasus, Paulowina imperialis, Prunus
persica, Paonia Moutan, Sterculia plantanifolia, some species
of Bambusa, &c. It frequents mostly those mulberry trees
which are thickly planted, or else those which are planted in
shady places, which are not directly exposed to sunshine or
wind. It may be found parasitic everywhere on stems or
branches of mulberry trees, but it most frequently attacks the
lower parts of the stem, a few inches or rather more above the
ground. The presence of the parasite as mentioned before, can
easily be recognized at a certain season by a snowy white ap-
pearance on the stems or branches, which are thickly covered
with the cocoons of the male scale insects. On the contrary

ON THE SCALE INSECT OF MULBERRY TREES. r2r

although the scales of the female insects remain throughout the
year, they can not easily be seen on account of their dull brown-
ish grey color, which is very similar to that of the bark on which
they remain tightly attached.

When the scale insects are found parasitic in swarms or lots
on stems or branches, the growth of the latter is more or less
interrupted by the loss of sap, as they may be weakened to a
certain extent, and particularly much more injury is done to the
younger stems or shoots than to the older ones, as the former
most frequently perish. Great as is the damage done by the
scale insects to the mulberry trees, few owners of trees take any
particular pains to destroy these formidable pests. The only
method commonly practised is to scrape off the scales from the
bark with small thin pieces of wood or bamboo, or sometimes
with the shells of ear-shells or mussels.

The preventive methods which I consider most efficacious
and most practicable for destroying this pest, is the spraying of
lime water, petroleum, or a mixture of water, fish-oil, and
bicarbonate of soda in the following proportion :-—

1. Water 1 litre
2. Fish oil 32 gr.
3. Bicarbonate of Soda 32 gr.

These preventive mixtures are more efficacious when applied
in dry weather during the months of June to October; for
during these months, the active young larve mostly distribute
themselves by crawling about, or remain fixed on the bark
without protection.

The method of the propagation of the insects are not as
yet clearly known, but most probably it may be done during the
first larval stage by the wind.

There is two natural enemies of the scale insect, one being
a hymenopterous insect belonging to Chalcide and the other a
coleoptera belonging to Coccinellide. Both of these decrease the
number of the pest every year to a certain extent.

Tokyo, January 1894.

LIST. OF REFERENCES,

No. 1. Laboratoire d’Etudes de la Soie. 1892.

No. 2. The Entomologist’s Monthly Magazine. Vol. 25.

No. 3. Bulletin de la Societé Entomologique Italienne, XXI
année, 1889.

EXPLANATION OF PLATES.

All the figures are drawn by C. Sasaki, except one
by the artist K. Yokoyama.

PLATE I.
Fig,.1. Male insect. Dorsal view. AOC) © .. 274;
eae 3 Ventral ,, ra
1, b. Head of - Dorsal: >; D. 2c! t
TotC, a Ventral ,, _
1, d. Antenne of ,, 13; OC. E
Te. Wee 9 D. oc. £
1, f. Balancer. -
Fig. 2. Female insect. Dorsal view. ASoces:
Bie: 3: 8 Ventral ,, So
Fig. 4. Pygidium. Dorsal ,, Doe. z
Fig. 5. 5 Ventral ,, -

areas.

Fig. 6. Antenne of female insect.

Fig. 7. Mouth parts of i

Fig.-8. Spiracle on the rst segment.

8, a. spiracle on the 3rd segment.

Fig. 9. Scale of female insect; a, moulted
skin of larva; b, moulted skin
of pupa.

Fig. 10. Ovipositor projecting out on the
ventral side of pygidium high-
ly mag.

Fig. 11. Eggs.

Fig. 12. | Newly hatched larva. Dorsal view.

i252 Pr Ventral ,,

12, b: Leg of ,,

12, c. Markings on the skin of §

PLATE II.

Fig. 13. Larva of 2nd stage. Dorsal view.

Fig. 14. Pe Ventral 5, ;
sp, Spiracle.

Fig. 15. Silky threads spun out from secre-
tory pores on the dorsal side.

15, a. Hemispherical pieces.

Fig. 16. Earlier form of male pupa. Dorsal
view.

16, a. Earlier form of male pupa. Ven-
tral view.

16, b. Earlier form of male pupa, high-
ly mag. Light spots dorsally,
shaded spots ventrally opening
pores.

Fig. 17. | Cocoon of male insect.

Fig. 18. | Cocoon with developed pupa within.

Fig. 19. Male pupa. Dorsal view.

Fig. 20. = Ventral ,,

EXPLANATION OF PLATES.

5,a. A group of circular or polygonal

99

Ll

123

124

Fig.

Fig.
Fig.

Fig.

aI.

22.

2a.

24.

EXPLANATION OF PLATES.

Branch of mulberry tree bearing
male cocoons and _ female
scales in masses.

Female pupa taken out of a scale.
Ventral view.

Posterior portion of female pupa.
Dorsal view.

Posterior portion of female pupa.
Ventral view.

Bull. Agric. Coll. Vol. Il. Pl. I.

" C. Sasaki del.

Bull, Agrit. Coll. Vol. I. Ph. I.

Fig. 16,0

(*

Al , pms
fai bal GE

K. Yokoyama del,

_C. Sasaki dei.

es

On the Spermatogenesis of the Silk-Worm.‘”

BY

Kametaro Toyama, Assistant in the Zoological Institute,
Agricultural College.

With Plates III—IV.

The work which forms the subject of the following pages
was carried on in the Zoological Institute of the Agricultural
College during last year. My original intention was to determine
the question, whether Verson’s large cell in the blind end of the
testicular follicle is truly a genital cell or not, and in studying
this I also touched upon the entire developmental history of
the spermatozoa as well as the most interesting question in the
development of genital cells, namely: the reduction of the
chromosomes.

Before going any further, I have, in the first place, to
express my sincere gratitude to Prof. Ishikawa for his kind
guidance and friendly counsel throughout the progress of my
work in his laboratory. I am also very much indebted to Prof.
Mitsukuri of the Science College for his kindness in allowing me
to use the library of the Science College, and also for various
other matters. To Prof. Sasaki I am also much indebted for the
materials. Thanks are also due to the director of the College,
Prof. Matsui who allowed me to continue my work in the College
after I passed through the university courses.

Methods of Investigation. The entire process of the spermato-
genesis was studied by means of sections and teased preparations,
the former method affording more advantages than the latter for
the researches of the division of the genital elements.

For teasing the testicular follicles, I used acetic acid
methylgreen for fixing and staining. As this causes the swell-
ing of the chromosomes, it is very useful in calculating their
number.

The following fluids are used in hardening the sections:
picro-acetic acid, sublimate alcohol, salt sublimate, chromo-

(1) The preliminary note of this paper was published in the ‘ Zoologischen
Anzeiger” No. 438 this year.

126 ON THE SPERMATOGENESIS OF THE SILK-WORM.

formic acid, Rabl’s platinum chloride, chromo-platinum chlo-
ride, 1% formic acid, and Flemming’s chromo-osmium acetic acid.

Of all these Flemming’s strong solution gives excellent
results on fixing the nuclear elements and the cytoplasm.

For staining the nuclear elements Flemming’s anilin-water
safranin solution, and Hermann’s triple staining is mostly
employed, while the cytoplasm is best stained by B6dhmer’s
hematoxylin. Most of my figures are, therefore, drawn from
preparations fixed by Flemming’s solution and stained by
these reagents. For killing embryos 1% formic acid solution
gives the best result. It not only preserves genital elements ex-
cellently, but other tissues are also very well preserved. Picro-
acetic acid is not very bad, but it causes the swelling of the
chromosomes, sometimes so intensely that it is impossible to
calculate their number; especially the longitudinal splitting of
chromosomes is utterly destroyed by its use.

For mounting sections, I used Gulland’s method, as it is
modified by Mr. S. Ikeda formerly assistant in the laboratory.
This is as follows :—

A very thin and even layer of the fixative (Mayer’s albumen)
is painted on the slide and a little distilled water is poured
upon it. The sections are then placed on the slide, the
excess of the water is wiped off with blotting paper and the
slide is warmed in an oven of about 30-35°C. until the water
completely evaporates. After this, it is treated as usual,
mounted and stained. By this method there is no need of a
section-smoother, as the rolling of sections may be completely
prevented and the position of sections may be changed at will
while we are placing them on a slide. In Gulland’s method,
the same effect may be obtained, but the fixing of sections on
a slide is not so strong, so that sometimes sections are removy-
ed when treated with alcohol, but in this method such a risk
is avoided.

Before going into the description of the spermatogenesis,
let us briefly describe the structure of the genital organs of
the silk-worm in the larval stage, although the same subject
has been treated of by other authors such as Cornalia (5),
Verson (38, 39), and Haberlandt (11).

The testes are paired, kidney-shaped, and lie at the right

ON THE SPERMATOGENESIS OF THE SILK-WORM. 127

and the left side of the dorsal vessel over the alimentary
canal, in the segment where the sixth stigmata opens (namely
eighth segment) and are firmly attached to the body-wall by
the trachea of the sixth stigmata and fatty tissues. As in
other Lepidoptera, each testis consists of four blind tubes or
testicular follicles, which are covered with a common envelope,
tunica adventitia, and each is placed facing the other with
the concave side, on both sides of the dorsal vessel. From
the middle of the concave side of each of these testes one
vas deferens arises, and the testicular follicles enter into it.
These ducts run, at first, along the dorsal side of the body,
gradually changing their course into the ventral side, and
finally attach themselves with a uterus-shaped process (fig 3)
found on the ventral median side of the twelfth segment. This
process seems to be changed into seminal vesicle, accessory
glands and ductus ejaculatoris.

In the female organs, all the relations are nearly similar
to that of the male except the shape of the ovary and the
mode of the attachment of the oviducts with it. The ovary
of the silk-worm is smaller than the testis from the early
beginning till to the last of the larval stage. It is somewhat
triangular in shape, and is situated each side facing the other
with a side of a triangle, the angles opposite to these sides
being produced to form the oviducts.

This is the usual form of the genital organ of Bombyx
“ mori in its larval stage (Fig. 1—z2). Cases are, however,
met with where the vas deferens arises from the outer side
of the testes as in the ovary, or one of the testes with its
vas deferens on its inner side and the other on the outer side.

Terminology. ‘The question of nomenclature presents some
difficulty, since so many different names have been given to
the same elements by different observers. So far as I could,
I have tried to avoid the use of such general terms as ‘‘ sper-
matoblast,” “‘spermatocyte,” “spermatogone,’ etc. and have
substituted simple descriptive expressions, as has been done by
O. Hertwig, C. Ishikawa, and vom Rath. We may distinguish
four stages in the sperm-formation of Bombyx mori. The
first of which, we may call the formative stage (‘‘ Keimzone”’ of
German authors), the second the growing stage (‘‘ Waschsthums-

128 ON THE SPERMATOGENESIS OF THE SILK-WORM.

periode”’ of German authors), the third the ripening stage
(‘‘ Reifungsperiode’’) and the fourth the stage of metamorphosis
(‘* Umwandlungsperiode”’). The cells of the first stage will be
called the primary germ-cells (Ursamenzellen), the cells of the
second stage, the speym-mother-cells after O. Hertwig. The cells
which divide two times successively in the ripening stage,
may be called the spevm-daughter-cells. This sperm-daughter-
cell changes itself into a spevmatozoon without further division.

THE DEVELOPMENT OF THE GENITAL ELEMENTS.

On examining a testis of a larva after the fourth moult,
we are struck with the varieties of cellular elements found in it.
As in the genital follicles of other Lepidoptera, the more
developed elements always lie near the vas deferens, while the
younger ones lie near the blind end of the follicle. In the
centre of the younger elements, near the blind end of each
testicular follicle, we find a large cell around which these
younger elements are arranged concentrically (fig. 7).

Before passing to a description of the development of the
genital elements, I shall say a few words as to the nature
of this large cell in the blind end, which I shall call
‘*Veyson’s cell.” As Verson has already worked over the
spermatogenesis of the silk-worm and described in two papers,
‘‘la spermatogenesi nel Bombyx mori; Padova, 1889,” and
‘Zur Spermatogenesis”’ in the Zoologischer Anzeiger, it is
necessary to quote here his opinion about this large cell.
His interpretation of it, as is given in the latter of these
works, is as follows:

‘‘In jedem Fache befindet sich nur eine einzige, grosse
Keimzelle; und aus dieser nehmen nach und nach alle or-
ganisirten Bildungen ihren Ursprung, aus welchen der Inhalt
des ganzen Faches besteht.”

Let us now see whether this interpretation of Verson corres-
ponds with my observations or not.

In the embryonic stage of the silk-worm, each testis
consists of only one follicle (Fig. 4) within which are scattered
round cells with distinct chromosomes and a nucleolus.

ON THE SPERMATOGENESIS OF THE SILK-WORM. 129

It is certain that these round cells in the testicular follicle
are genital cells. When the larva is about to be hatched, three
depressions appear on the follicular wall. These gradually
deepen until four cavities are formed. This is shown in
fig. 5 f. Hand in hand with this change another depression
appears on each of these testicular tubes as fig. 5 7 shows, and
in a testis of a larva four days old, there is seen a large cell
in each of these secondary depressions of the follicle. This
large cell is the origin of Verson’s cell found in the blind end
of the testicular follicle. Similar structure is also found in a
testis a little younger than this, but here a distinct membrane
exists between the depression and the follicular space (fig. 6),
so that the depression is produced by the swelling in of one of
the cells of the follicular wall, and has no connection with
the genital cells lying in the follicular space. It is therefore
certain that Verson’s cell is derived from one of the follicular
cells, and not from the genital cells.

This depressed cell gradually loses its membrane and pro-
duces amoeboid processes between the genital cells (fig. 11). It
always has an elliptical nucleus which contains many chromatin
granules (figs. 9, 10, II, 12, 13, v), staining very deeply by such
reagents as hematoxylin, carmine, and aniline dyes, and is
connected with the follicular wall by a protoplasmic strand
(fig. 8, p). It thus resembles very much the nucleus of follicular
cells (fig. 6) which also contains many chromatin granules.
Contrary to the assumption of Verson it has no nucleolus, nor
any karyokinetic division to be found in it at any stage of its
existence from the early beginning till the death of the moth.
Verson’s cell is, as stated above, generally situated in the blind
end of the testicular follicle, around which the youngest sexual
cells are found (fig 7 v). Sometimes, however, it happens to be
placed at one side of the follicle (fig. 10 v) where more developed
genital cells are found, the most interesting thing about here is
that the youngest genital cells are here also found at the blind
end and not around Verson’s cell. As will be seen in fig. 10 the
genital cells at the blind end of the follicle divide profusely as
usual, although Verson’s cell is not present, and this shows that
Verson’s cell has nothing to do with the formation of genital cells.

The division of Verson’s cell takes place, as Verson states,

130 ON THE SPERMATOGENESIS OF THE SILK-WORM.

amitotically. Its nucleus becomes constricted into two or more
portions producing from one to many small nuclei on the side of
the large one. This mode of division is not to be seen in the
first larval stages, but commencing generally at the end of it,
gradually increasing to the imaginal stage (Figs. 6, 11, 9, 12, 13 v);
but the genital elements are seen to increase in number from the
first larval stages, where no division of Verson’s cell is found.

In a testis of an imago after copulation, I have also met with
Verson’s cell, but now it does not stain so well by the reagents
above described as in earlier stages (fig. 13). At this stage, there
is seen a number of cells close to Verson’s cell, probably corres-
ponding with the cells formed by the above described amitotic
division of it. These cells also take no stains and contain large
vacuoles, showing thus the process of degeneration.

From all this we arrive at the conclusion that Verson’s cell
is not a genital cell as Verson states, but it is a supporting cell
connecting all the younger genital elements with the wall of the
testicular follicle and probably nourishing them. This confirms
the assumption of Ziegler and vom Rath (46), who says that ‘‘es
erscheint eine Deutung zulassig, welche die Befunde von Verson
mit denen von vom Rath in Uebereinstimmung bringen konnte,
ndmlich die Auffassung, dass die kleinen Zellen nicht die Ab-
k6mmlinge, sondern sozusagen die Geschwister der grossen Zelle
(Verson’s cell) sind und dass sie durch successive mitotische
Theilung die zahlreichen Samenbildungszellen ‘erzeugen, wahr-
end der Kern der grossen Zelle, welche den Character einer
Rand- oder Stiitz-zelle hat, mehrfach sich amitotisch theilt.”’

Verson’s cell may therefore be safely assumed to be a
supporting cell of the testicular follicle, and is also to be seen
in the blind end of an egg tube as is shown in fig. 14 and in quite
young stages, it is very difficult to distinguish the male and
the female elements except by the external shape of the follicle
as already described.

(t) The cells which divide amitotically in the genital follicles of the silk-worm
do not belong to the cycle of sexual cells as above described and this confirms the
opinion of vom Rath (29, Il) who says that “‘Dem Mitosen gegeniiber haben die
Amitosen durchweg einen mehr oder weniger deutlich erkennbaren degenerativen
Character. Die Mitose hat sich keineswegs aus der Amitose entwickelt, so dass die
letztere den urspriinglicheren Theilungsmodus darstellte.”’

ON THE SPERMATOGENESIS OF THE SILK-WORM. I31I

We have also met with a similar large cell in the testes of
the wild silk-worm (fig. 16), Papilio xuthus L. (fig 15), P. mach-
aon, L., P. alcinous, Klug., but not in Antherea yamamai, Guér.
Mén., Caligura japonica, Moore., Rhodia fugax, Butl., etc. So
it seems evident that in the genital glands of Lepidoptera, such
a cell connecting the younger genital elements with the follicular
wall, aithough not constant, is not of rare occurrence. (

I. The Formative Stage. The genital cells of this stage or the
primary germ-cells, are situated near the blind end of each tes-
ticular follicle and are arranged concentrically round the sup-
porting cell as already mentioned (figs. 7, 11). The youngest
primary germ-cells are round with distinct chromosomes and
nucleolus (fig. 4). When the supporting cell enters the testicular
follicle and becomes connected with the genital cells, the latter
assume a conical shape (figs. 11, 18), the apices of which are
connected with the protoplasmic processes of the supporting cell.
The nucleus of these germ-cells is always situated at the basal
part of the cell, corresponding exactly to the young genital
cells of Pyrrhocoris apterus described by Henking (15). In each
follicle are seen many generations of the primary germ-cells
showing the various stages of typical karyokinesis. In figs. 17-22
are represented various stages of division of primary germ-cells.
Fig. 17 and 18 show cells in the resting stage; their nuclei
present a spherical shape with a distinct nuclear wall.
Chromatins are scattered in fine reticulum. Nucleolus is
clearly to be seen in the centre of the nucleus. In a pre-
paration fixed by the Flemming’s strong chrom-osmium acetic
acid solution and stained by Hermann’s safranin-gentiana-orange
the nucleolus is found to be suspended in the chromatin-net
as one or two globules, and consists of small round chro-
matic bodies.

In the nucleus represented by fig. 17, is seen only a
single nucleolus in the matrix of which small round granules
are clearly to be seen, while in other cells (fig. 18), the

(1) Cholodkowsky’s (6) observation on the testis of a Diptera, also confirms
the presence of such a large cell in the follicle, but it seems to be of an entirely
different nature from that of the silk-worm, as it divides mitotically.

132 ON THE SPERMATOGENESIS OF THE SILK-WORM.

nucleolus consists of four somewhat large chromatic bodies.
These chromatic bodies arrange themselves in single or double
rows, or sometimes very irregularly, Henking (15) also pointed
out a case somewhat resembling this in a nucleolus of Pyrrho-
coris apterus.

When the primary germ-cells begin to divide, the fine
net-work of chromosomes gradually becomes coarser (fig 19), and
sometime after a beautiful skein stage is to be seen. I was
not able to find, at this stage, the longitudinal splitting of the
chromosomes, but this is clearly to be observed in such cells in
which the nuclear segments can be distinctly made out (fig. 20).

This corresponds to the segmented skein of Flemming.
Although the chromosomes of the primary germ-cells have
divided longitudinally in this stage, they do not separate
from each other until they form the equatorial plate. The
double chromosomes thus formed present the appearance of
being only one when viewed from one side (fig. 21). When the
division proceeds a little further, the chromosomes separate from
each other and go to the poles (fig. 22). Owing to the small
size of the primary germ-cells, the exact number of the
chromosomes can not be made out. I was, however, enabled
to count, in favorable specimens, twenty-six to twenty-eight
chromosomes in polar views.

The same mode of division, as now described, takes place
many times in this stage, and the primary germ-cells become
at last reduced to about two-thirds or less of their original size.

II. The Growing Stage. In the first part of this stage,
the genital elements are small, in consequence of the repeated
karyokinetic divisions of the primary germ-cells. I call these
cells by the name of sperm-mother-cells (‘‘Samenmutterzel-
len”). In the resting stage (fig. 23), the sperm-mother-cells
are similar in appearance to that of the primary germ-cells.
A nucleolus is generally seen in the net-work of linin and
chromatin.

The sperm-mother-cells gradually enlarge, and their nuclei
go through marked changes (figs. 24-46).

Most of the chromatin granules become collected to one
side of the nucleus, and form an irregular mass. Fine linin

ON THE SPERMATOGENESIS OF THE SILK-WORM. 133

fibres with scanty chromatin granules are seen radiating from
the mass to the wall of the nucleus. This is shown in figs. 24-26.

The nucleolus, lying either in the chromatin mass or out-
side of it, persists, as is unusual in skein stages of other animals,
till to the end of the skein stage shortly to be described.

The chromatin granules once collected into a single irregu-
lar mass, become again separate from each other and arrange
themselves along the radiating linin fibres, and the skein stage
is thus obtained (fig. 27). In the nucleus of this stage we can
easily observe Rabl’s pole-field.

The same change of nucleus of the beginning of the growing
stage was observed by Brauer (4) in Ascaris megalocephala
where the longitudinal splitting of chromosomes was also
observed. I have carefully searched after this in Bombyx, but
could not find it in any of the skein stages.

The most singular thing about here is that the chromatin
substances once more go through changes similar to those
now described. In fig. 28 the chromosomes again have lost
their even outline, and present a granular appearance. They
became moreover much shorter than before. These granulations
soon become more defined, and their lineal arrangement is
gradually destroyed until the nucleus presents the appearance
shown in figs. 28’, 29. The nucleolus, however, shows no change
from the first resting stage till the present stage, and always
consists of small chromatic granules imbedded ina less stainable
matrix (figs. 23-29). Hand in hand with these changes of the
nuclear substances, the cell-body gradually enlarges, but no
metamorphosis of the cytoplasm is as yet to be recognized.

The chromatin granules scattered in the nucleus become
again collected in the centre of it and present an irregular mass
as before. ‘The position of a nucleolus, or two nucleoli, is also
either in the centre of the mass of the chromatin granules or
outside of it in the cavity of the nucleus (fig. 30).

The same structure of the sperm-mother-cells as now de-
scribed is also found in Papilio xuthus, P. alcinous, Calygura
japonica, and Rodia fugax.

In a still later stage, the chromatin granules again com-
mence to separate from one another, and the nucleus again pre-
sents the appearance shown in figs. 31 and 32. In most cases

I34 ON THE SPERMATOGENESIS OF THE SILK-WORM.

two nucleoli are found in the nucleus of this stage, these
gradually migrate towards the periphery of the nucleus facing the
centre of the cyste (rarely, facing the wall of the cyste) and are
finally pushed out into the cytoplasm one after the other through
the nuclear wall at this point (figs. 33-38, c,d). Placed in the
cytoplasm the nucleoli seem to change their quality, since they
now stain differently from what they did when they were in the
nucleus. This is shown by the use of Hermann’s triple stain-
ing, by which the nucleolus in the cytoplasm takes a brownish
colour, while it colours deep red so long as it is within the nucleus.
The further fate of the nucleoli in the cytoplasm is not known.

After the disappearance of the nucleoli from the nucleus,
two attraction-spheres make their appearance in the cytoplasm
just at the same place where they disappeared. These two
attraction-spheres gradually recede from one another until they
come to the opposite poles of the nucleus. Some faint achro-
matic fibres are seen running between these two attraction-
spheres at this time (fig. 48).

Before the extrusion of the two nucleoli from the nucleus,
we can distinctly see in the cytoplasm between the nucleus and
the wall of the cyste, an aggregation of microsomes (fig. 40),
which in all appearance are like the ‘“‘ Nebenkern”’ described by
la Valette St. George in a testis of Forficula auricularis (36) and
others. This aggregation of cytomicrosomes has not always the
same appearance in the sexual cells of the same cyste, and some
of them present fibrous structures very similar to those of the
achromatic spindle (figs. 36, 41-43, 47). As shown in figs. 36 and
41 this fibrous bundle is seen at first parallel to the base ofa cell.

As Henking (15, 16) and others have already shown in other
animals, all the genital cells of Bombyx mori m the same cyste
are nearly in the same stage of development. Consequently
we may say that the aggregation of cytomicrosomes found in
one cell and the fibrous bundle in the other are two successive
stages, the former changing into the latter.

The fibrous bundle, above mentioned, takes somewhat oblique
position and gradually approaches the nucleus, and, when the at-
traction-spheres come to lie in the opposite sides of the nucleus,
it connects with them forming a spindle (figs. 42, 43, 47, 49).
The fibrous bundle, or the spindle, thus formed (fig. 49) consists

ON THE SPERMATOGENESIS OF THE SILK-WORM. 135

of three parts, the two polar parts which stain faintly with
hzmatoxylin and the median part which stains deeply with the
same dye.

Beside this change there is to be seen a deep colouring spot
surrounded by a free area either within the cytoplasm of sperm-
mother-cells or in the boundary between two such cells. It is
either round or ellipsoidal in shape, and sometimes it has a con-
striction in its centre (fig. 33, v) giving it the appearance of a
dumb-bell (figs. 32-37 v, 39-43 ¥, 47 v). This is surely the “ Ver-
bindungsbriicken” of Platner (27). Nothing can be said of its
first appearance or its disappearance, but as it is found in cells
in which the centrosomes are already present, it can not be the
centrosome, although it appears very much like it, especially
when it is found in the cytoplasm.

A little before the appearance of the centrosomes in sperm-
mother-cells the chromatin granules as shown in fig. 44 gra-
dually collect here and there and assume ring-shaped struc-
tures. These rings have already been described by Henking (15)
and vom Rath. (28). Each of these rings again dissolves into
four small chromatin granules (fig. 45) as vom Rath observed in
a sperm-mother-cell of Gryllotalpa and Salamandra (28, 29).
After the breaking up of the chromatin ring, the separated
chromosomes are now ready to divide.

In the preparation fixed by Flemming’s strong chrom-
osmium acetic acid, the ‘‘ Verbindungsbriicken”’ is clearly to
be seen without any staining, while the spindle fibres can not
be seen unless they are stained by hematoxylin. I am not able
to observe these structures in the preparation fixed by picro-
acetic acid and stained by picro-carmine.

In the sperm-mother-cells of the silk-worm, no accumula-
tion of yolk granules is to be observed in the cell-body, although
in other animals, such as in Ascaris and in Pyrrhocoris, this is
the case.

III. The Ripening Stage. In this stage, we find fully grown
sperm-mother-cells each of which divides two times succes-
sively and produces four cells which I will call the sperm-
daughter-cells. These change themselves, without any further
division, into spermatozoa.

136 ON THE SPERMATOGENESIS OF THE SIKL-WORM.

The sperm-mother-cells with large chromatin granules and
distinct centrosomes, described in the last part of the grow-
ing stage, now entirely lose théir nuclear wall and the chro-
mosomes become arranged in an equator of the spindle form-
ing a ‘‘Kernplatte” of Strasburger. In this stage, I have
not found any nucleolus in the ‘“ Kernplatte,” while Hen-
king observed it in a spermatocyte of Pyrrhocoris apterus.
Fig. 50 shows a side view of this stage. The chromosomes are
arranged in a single row in the equator of the spindle, and
not in double rows as Henking (15, 16) and vom Rath (28) ob-
served in genital cells of Gryllotalpa, Pyrrhocoris and Pygaera.
In a polar view of ‘“ Kernplatte,”
twenty-eight chromosomes (fig. 52) in most of the cells, although
some with twenty-six or twenty-seven are sometimes to be found.
Each of the chromosomes of the ‘‘ Kernplatte,” gradually pro-
duces a constriction in the middle of the long axis and forms a
dumb-bell-shaped chromosome (figs. 50’, 51). This constriction
deepens until two chromosomes are formed. This mode of divi-
sion corresponds very well with the division of sperm-mother-
cells of Diaptomus described by Ishikawa in the following words:
“(In the process of division each dumb-bell-shaped chromosome
elongates and becomes divided in its middle part, so that one
half of the dumb-bell goes to one pole and the other half to the
other. The only difference from the ordinary karyokinesis con-
sists in the mode of division of the chromosomes, which general-
ly divide longitudinally and not transversely.”” In fig. 50’ isa
‘‘Kernplatte”’ of a cell in which the process of the transverse
division of the chromosomes is to be seen. This is certainly an
intermediate stage which is described in fig. 50 and fig. 51. After
the transverse division, each row of chromosomes gradually re-
cedes to the pole and forms a diaster stage (fig 53). This corres-
ponds to the first reducing division of Wetsmann, each daughter
nucleus containing also twenty-eight chromosomes as the mother-
cells. Consequently, in this division no reduction of the number
of chromosomes takes place, as Henking observed in the egg-and
sperm-cells of many other animals (15, 16), while it corresponds
very well with the first division ofa sperm-mother-cell of Ascaris
and Gryllotalpa described by O. Hertwig (18) and vom Rath (28).

After the division the chromosomes form a somewhat granular

we can clearly distinguish

ON THE SPERMATOGENESIS OF THE SILK-WORM. 16e}9/

mass collecting in the centre of a nucleus, but no resting stage
is formed. The fate of the centrosomes is not known. ‘The
collected chromosomes gradually separate from one another and
form a loose mass of chromatin granules.

A full grown spindle with a “ Kernplatte”’ is again formed.
Owing to the reduced size of the cell-body after the first division
(compare figs. 51 and 52 with figs. 54 and 55 all of which are
magnified to the same diameter), the spindle together with that
of the individual chromosome is very small in this spindle, so
that we can easily distinguish it from the spindle of the first
division. A side view ofa cell in this second division is shown
in fig. 54. In this division, the chromosomes are arranged in a
single row, and no constriction is to be seen in any of them. In
the polar view of the ‘‘ Kernplatte,” twenty-eight chromosomes
can be counted (fig. 55). These separate into two groups and
form the chromosomes of sperm-daughter-cells, each of which
therefore contains fourteen chromosomes. Fig. 56 represents the
diaster of this stage. The exact number of the chromosomes
can not be made out in the side view, but in the polar view we can
clearly count fourteen (fig. 58). Sometimes, a darker structure,
somewhat resembling the ‘‘ Verbindungsbriicken,” is to be seen
under the polar spindle (fig. 57, lower figure), but I am not able
to tell what this really represents.

A single or a double row of cell-plates appear at the equator
of the ‘‘ Verbindungsfaden” (Figs. 58, 59, 60) and the cell be-
comes constricted. The central and the polar spindles can now
be clearly distinguished from one another (Fig 59). The former
appears as a compact mass presenting a median darkly stainable
part, while the polar spindle forms a somewhat semicircular
ring having a centrosome in its centre.

The daughter-cells, after these changes, completely separate
from each other. These we call sperm-daughter-cells which
correspond with the ‘‘Spermatides” of Ja Valette St. George.

IV. The Stage of Metamorphosis. In this stage, the sperm-
daughter-cells gradually change themselves into spermatozoa. In
figs. 61 and 62 we have five sperm-daughter-cells. The ‘‘ Verbin-
dungsfaden” gradually contracts, accumulating in its end some
microsomes (fig. 62 a,b). These ‘‘ Verbindungsfaden ”’ become
gradually coarser, as is represented by fig. 62 c, m, and finally

138 ON THE SPERMATOGENESIS OF THE SILK-WORM.

change so as to form a granulated ball with a free plasm around
them (fig. 62d, ). Along with these changes, the chromosomes
gradually separate from one another and arrange themselves at
the periphery of the nucleus. Figs. 63 and 64 are drawn from
fresh specimens treated with acetic acid methyl-green. Chromo-
somes collect near the nuclear wall and present a moniliform
appearance in a side view. The fully grown ‘‘ Nebenkern”’ (n)
and mitosomes (m) are seen, one of the latter surrounded by a
free plasm. In fig. 64 beside the ‘‘ Nebenkern”’ and mitosomes,
a row of microsomes is to be seen running through the axis of
the tail. A further stage with a more elongated head and a
pretty long tail is represented in figs. 65-83.

In fig. 65 is given the head of a spermatozoon at a some-
what advanced stage. Chromosomes are as in the first stage
arranged at the periphery of the nucleus, while the ‘ Neben-
kern” appears more compact than before, slightly presenting
its granular structure. The position of mitosomes is very irregu-
lar, being placed either below or above the ‘‘ Nebenkern”’ (figs.
65, 66 b, 68, 70, 71).

Sometimes we see that it is situated at the extreme point of
the head of a spermatozoon anterior to the nucleus (fig. 67 m).

Beside these structures there are to be seen some other
granulated spots, which are sometimes coagulated into a single
mass, sometimes scattered more irregularly (figs. 65, 68, 71,
72). These phenomena are very similar to those described by
Henking in Pyrrhocoris (25). Gradually these ‘‘ Nebenkern”
elongate into the structure of the tail-part (figs. 68, 69, 70, 71,
72, 2). In the cross section of the tail passing through the
‘* Nebenkern” of this stage, radial processes are seen from the
“Nebenkern”’ to the wall of the tail of the spermatozoa (fig.
70 b, E-2).

The ‘‘Nebenkern” elongates more and more, becoming thin-
ner, and with this change, the mitosome gradually becomes fainter
and smaller. The radial processes of the ‘‘ Nebenkern”’ disappear
(figs. 73, 77). In fig. 73 a cross section of the median part of
the tail with a ‘“‘ Nebenkern”’ in the centre of it, is represented.

Hand in hand with this change, chromosomes gradually
accumulate at the one side of the periphery of the nucleus. This
accumulation presents a somewhat crescent shaped figure, but the

ON THE SPERMATOGENESIS OF THE SILK-WORM. 139

individual chromosomes are still to be recognized (fig. 75 a, 76).
An entire spermatozoon of this stage is represented in fig. 83.
Here the ‘“‘Nebenkern”’ is no more to be seen as a distinct
body as before, but appears only as a thicker mass with a fine
thread-like prolongation. The mitosome has also completely
vanished from sight.

Figs. 78, 79 and 80 show more advanced stages. The
chromosomes which are accumulated at the periphery of the
nucleus became now coagulated into a single compact mass and
form a deeply stainable body with a pointed end and a vacuole
in the centre of it.

In fig. 82 is shown a spermatocyste with nearly matured
spermatozoa contained in it. In the head part of the cyste, is
seen a large cell metamorphosed from a nucleus of the cyste and
functioning as a supporting cell.@) The form of this cell is very
similar to the large supporting cell in the blind end of the
testicular follicle and may be compared with the cells des-
cribed and figured by Flemming in a spermatocyste of Sala-
mandra maculosa (9).

ON THE ‘‘ HODENZWISCHENKORPERCHEN.”’

The ‘‘ Hodenzwischenkérperchen”’ of van Beneden are also
to be found in the genital follicles of the silk-worm of both
sexes. In preparations fixed by Flemming’s strong solution
and stained by Hermann’s triple staining very conspicuous cells
are to be seen here and there (figs. 7h, 84). They are round
with homogeneous protoplasm, in the centre or at the periphery
of which a deep stainable chromatin body, sometimes with va-
cuoles in it, is to be seen. Beside this, sometimes one or
more small deeply stainable spots occur in the cytoplasm. In
the early part of the formative stage, these cells are found
in the midst of the primary germ-cells, while in later stages
the entire mass of cells in a cyste presents such a structure
except those at the wall of the cyste (fig. 84). They are also
to be seen in the growing stage, where their structure is similar
to those of the formative stage.

Sometimes the granular stainable spots in the cytoplasm

(1) This is already described by Tichomiroff (37).

I40 ON THE SPERMATOGENESIS OF THE SILK-WORM.

increase in number and in the centre of them a large chromatin
body is to be seen (fig. 85).

In other cells the central chromatin body is very much
reduced in size, and the smaller granules increase profusely.
In more advanced stages the outline of the individual ‘‘ Hoden-
zwischenkorperchen ” has disappeared and at last we see a cyste
which contains only granules (fig. 86). Such cells are more
abundant in a testis or an ovary attacked by Nosema bombysis
than in healthy ones.

These cells are certainly similar to ‘‘globules residule’
(van Beneden) or “‘ Hodenzwischenkorperchen” (O. Hertwig) oi
the testes of Ascaris megalocephala. Similar cells are also
described by Hermann and Flemming in the genital organ of
Salamandra. These authors come to the conclusion that they
are degenerating cells. The description above given confirms
this view and I am quite of the opinion of O. Hertwig when
he says (18):—‘‘die Zwischenkorperchen nichts anderes als
verkiimmerte, zu Grunde gehende Hodenzellen sind.”

COMPARISON OF VERSON’S OBSERVATIONS WITH MINE.

As Verson is the only naturalist who studied the spermato-
genesis of Bombyx more or less thoroughly we will here compare
the results obtained by this naturalist with mine.

His description, as given in Zool. Anz., is as follows :—

‘‘In einem bestimmten Entwicklungsstadium der Hoden
schliessen sich nun an die Keimzelle die Gebilde, die sich nach
einander beschreibe, in ununterbrochner Reichenfolge bis zur
Spitze des Hodenfaches, wo spater dieses letztere in das gemein-
schaftliche vas deferens aller vier Facher mit auswachsen wird.

‘‘r. Die friiher erwahnten Kerne haben sich aus dem Ver-
bande mit den strahlenartigen Fortsatzen der Keimzelle los-
gemacht; sie erscheinen nun selbstandig, und mit einem diinnen
Plasmahof umgeben.

‘‘2. Es folgen rundliche oder mehr unregelmdssige Proto-
plasma-klumpen, welche mehrere bis zahlreiche Kerne ent-
halten, und da die Kerne an masse vorwiegen, so nimmt der
ganze Klumpen eine Maulbeerform an.

ON THE SPERMATOGENESIS OF THE SILK-WORM. I4t

‘3, Groéssere, ungefahr spharische Klumpen, welche jedoch
viel heller und an der adusseren Peripherie durch eine reinere

Kreiscontour schon begrenzt erscheinen. Die Kerne sind eben-
falls hell, blaschenartig geworden, und anschliessen glanzende,
scharf unschriebene, im Profil komma- oder hufeisenformig
gestaltete Korperchen.

“4. Noch grodssere Blasen, an welchen eine Umhiillungs-
schicht und eine Inhalt unterschieden werden muss. Letzterer
ist durch Kerne. gegeben, die, viel zahlreicher als in No. 3, hier
und dort deutlich eine schmale umgebende Plasmaschicht erken-
nen lassen und sich von innen epithelienartig an die Umhiillungs-
schicht anlegen, wahrend gleichzeitig ein centraler Raum er-
scheint, welcher von geformten Elementen frei bleibt. An Grosse
und sonstigem Aussehen ahnliche Kerne erblickt man auch in
der Umhiillungsschicht der Blase, wenn auch sparlich vertheilt.

fies Ahnliche Blasen wie die unter 4 beschriebenen, nur von
grosserem Durchmesser. Demgemass sind auch die einzelnen
Elemente der inneren Auskleidung gewachsen, und besonders
deren Protoplasma viel breiter in’s Auge fallend. Die Kerne der
Umhiillungsschicht sind jetzt abgeplattet, und zeigen sich im
Durchschnitt spindelformig.

“6. Ahnlich grosse Blasen mit verdindertem Inhalt. Die
epithelartige Lagerung der enthaltenen Zellen ist verschwunden.
Letztere erscheinen mehrere Mal kleiner als in der vorher-
gehenden Reihe, und fiillen ohne Ordnung die ganze Hohlung
der Blase aus. Ihre Kerne schliessen haufig scharf markirte
Kernkorperchen ein, wie jene sub. 3 angefiihrten.

“‘7, Die Blasen dehnen sich ungleichformig und zwar nach
einer einzigen Richtung aus, so dass die spharische Form einer
birn-oder schlauchartigen zu weichen beginnt. Die Umhiil-
lungsschicht wird dabei noch diinner, und die Zellen des In-
haltes fangen im Mittelraume des Schlauches so zur zerfallen
an, dass die scharf markirten Kernkérperchen frei werden,
wahrend das Protoplasma sich in langlich ausgezogene Tropf-
chen auflést. Die peripheren noch nicht zerfallenden Zellen
stellen sich radiar zur Liangsachse des Schlauches auf, und
laufen zunachst gegen dieslbe zipfelig aus.

‘8, Langliche Schlauche, an deren stumpfen, abgerundetem
Ende die scharf markirten Kernkérperchen oder Derivate der-

I42 ON THE SPERMATOGENESIS OF THE SILK-WORM.

selben sich ansammeln, wahrend der iibrige Inhalt durch
langlich ausgezogene Trépfen gegeben ist, die sich zu varicésen
Faden anreichen.”

Let us now compare Verson’s results with mine above de-
scribed. The description in his first paragraph is certainly that
of my primary germ-cells. The second is also primary germ-
cells in their later stages. The third and fourth are the cells of
the growing stage and similar to my sperm-mother-cells. The
““komma- oder hufeisenformig gestaltete Korperchen” in the
nucleus are undoubtedly derived from a bad preservation of the
materials, and seem to be caused by coagulation of the chro-
matin granules in the granular stage of the nucleus in young
sperm-mother-cells. The fifth and sixth are the cells of the
ripening stage. Just, as Verson stated, the sperm-mother-cells
are generally arranged peripherally round the inner side of a
cyste in a row, while after the first division such an arrange-
ment is completely destroyed. Between the two successive
divisions of the sperm-mother-cells no nucleolus appears in a
nucleus. Consequently his description ‘‘ihre Kerne schliessen
haufig scharf markirte Kernkorperchen ein” seems to be due
to his mistaking the coagulated mass of chromosomes for the
nucleolus. The seventh and eighth are the stage of metamor-
phosis. In this stage, he also mistakes the coagulation of the
chromosomes for a nucleolus.

In general, his result agrees with that of my own investiga-
tion, though in the details it differs very much, probably in con-
sequence of his defective methods of investigation.

SPINDLE FIGURES, ‘‘ NEBENKERN,’ CENTROSOMES
AND NUCLEOLUS.

As to the origin of the achromatic spindle, many facts have
been observed by eminent naturalists such as van Beneden, (1)
Flemming (7), O. Hertwig (19), Platner (25), Strasburger (33, 34),
Hermann (17), Bovert (2), Brauer (4), Waldyer (40), Rabl, (29)
Watase (41) Schewiakoff (3), Ishikawa (21, 22) and many others,
in different animals and plants. In general, there are, as far as
my knowledge goes, three views as to its origin: (1) the achro-
matic threads arise chiefly from the cell protoplasm; (2) they

ON THE SPERMATOGENESIS OF THE SILK-WORM. 143

arise from the achromatic thread substance of the nucleus; (3)
they arise both from the achromatic nuclear constituents and
from the cytoplasm. O. Hertwig states in his celebrated text
book ‘“‘ Die Zelle und die Gewebe” that, “Ich have frtiher den
Standpunkt vertreten und nehme ihn auch jetzt noch ein, dass,
abgesehen von den Polstrahlungen, die dem Protoplasmakorper
der Zelle angehoren, die verschiedenen Structurtheile der Kern-
figur von den einzelnen Substanzen des ruhenden Kerns abstam-
men. Die stoffliche Grundlage fiir die Spindel und die spater
aus ihr hervorgehenden Verbindungsfaden suche ich in dem
Liningeriist.”’

In the sperm-mother-cells of Bombyx mori, it seems to be a
clear fact that the central spindle is derived from the cytoplasm
as will be seen in figs. 40, 41, 42, 43, 47, and 49. Of the polar
parts of the achromatic spindle, I am strongly inclined to believe
that they are derived from the nucleus, because at the time when
the central spindle is formed in the cytoplasm (figs. 36, 41, 42
and 43), both the centrosome and radial fibres are as yet not to
be seen. When centrosomes appear at the two poles of the
spindle, the radiating fibres come distinctly into view (fig. 49).

Let us now consider the origin of centrosomes. After the dis-
covery of this body by E. van Beneden in Ascaris megalocephala,
Flemming (8), Guignard (10), Strasburger (34), Hetdenhein (13)
vom Rath (29) and others have shown that they are present in
the cytoplasm also during the resting stage of the nucleus. I
have also found a distinct centrosome with a well developed ar-
choplasm in the resting sperm-mother-cells of Panulirus japonicus
(figs, 87, 88, 89 and go), but in the sperm-mother-cells of Bombyx
mori, I am unable to detect the centrosome in the resting stage.

Watase (42) considers the centrosome as an aggregation o
the cytomicrosomes, and states that “‘ for the centre of the aster
is the point where the greatest number of cytoplasmic filaments
meet with one another and the size of microsomes produced
at such a place must be correspondingly large. In other
words, the microsome produced in the centre of the aster is the

centrosome”.” A. Brauer (3, 4) discovered the formation of

1) Schneiders’s observation is little different from Watase’s. He states that the
attractions—spheres consist of a convolution of thin threads, and the radiated threads
form the ‘‘ Polsonne.”’

144 ON THE SPERMATOGENESIS OF THE SILK-WORM.

centrosome from a nuclear matter and states in his beau-
tiful work recently published ‘‘ Zur Kenntniss der Spermato-
> that ‘‘in dem einen Falle,
bei Univalens, tritt das Centrosom in Kerne auf, wachst und
theilt sich hier, und die Halften riicken dann nach zwei einander
entgegengesetzten Punkten auseinander und treten durch Liicken
der Membran in das Protoplasma tiber, die Spindelfasern gehen
aus den Fasern hervor, welche vom Auftreten des Centrosomes
an letzteres und das Chromosom verbinden, die Spindel ist
mithin in allen ihren Theilen eine einheitliche Bildung, sie
besteht nur aus Kernsubstanz”).”

In the testes of Bombyx mori as above stated the extruded
nucleoli are probably the origin of the centrosome. Although
I was not able to see exactly as Brauer did in the testes of
Ascaris, I can not help thinking that there must be some genetic
connection between these two bodies, there being such an exact
correspondence in the time and the place of the disappearance
of the nucleolus and the centrosome. The only difference
between them being the size, which is much greater in the
nucleolus than in the centrosome. This, however, is probably
caused by the formation of the polar spindle from a part
of the nucleolus. If this is proved, then the central spindle
is derived from the cytoplasm and the polar spindle from the
nucleus.

In the structure of the spindle, my results are entirely
similar to those of Heymann who says: ‘an der fertig ausgebil-
deten Spindel wird dieser Theil die axiale Mitte derselben ein-
nehmen, weshalb ich ihn mit dem Namen Centralspindel belegen
mochte und wird aus Fibrillen bestehen miissen, die direct und
continuirlich von Polkoérperchen zu Polk6rperchen ziehen, ohne
auf ihrem Wege itiberhaupt mit chromatischen Kernelementen
in Beziehung zu treten. Gewissermassen als Mantel werden
sich tiber diese Centralspindel jene Fasersysteme heriiberlegen,
die von den beiden Centrosomen aus zur Herbeiholung der
Chromatinelemente entsendet wurden, und diese Fibrillenziige

genese von Ascaris megalocephala,’

1) Wasielewski (43) also states the relation between centrosome and nucleus
in the genital cells of Ascaris megalocephala, while Karsten (23) has observed the
extrusion of nucleolus from a nucleus to form centrosome in cells of plants (Sporan-
gium of Psilotam).

ON THE SPERMATOGENESIS OF THE SILK-WORM. 145

k6nnen nicht von Pol zu Pol ziehen, sondern werden in der Nahe
des Spindelaquators durch ihren Ansatz an die sich farbenden
Kernbestandtheile eine Unterbrechnung erleiden miissen.”’

The “ Nebenkern” is according to la Vallette derived from
cytomicrosomes, where he says (35) “hier wie dort ganz
unzweifelhaft seine Entstehung aus dem kornigen Zellplasma zu
constatieren,” while Platncr (26) assigns its origin to the nu-
cleus. On this point he says: ‘‘ Hier entwickelt sich aus dem
Kern ein eigenthiimliches, an frischen Preparaten glanzendes,
durch dunkleres Aussehen und homogene Beschaffenheit von
dem umgebenden Protoplasma sich unterscheidendes Element.
Es ist der Nebenkern in seiner ersten Anlage.” Beside this
he (27) describes in the cytoplasm of the spermatocytes of
Lepidoptera, a structure which he designates by the name of
“* Verbindungsbriicken.”

In the sperm-mother-cells of the silk-worm, we also find a
body which has the same appearance as both Platner’s ‘‘ Verbin-
dungsbriicken”’ and his centrosome. As it is present in the
cytoplasm even after the appearance of two centrosomes, it is
not a centrosome, so I will call it the ‘‘ Verbindungsbriicken”
after Platner, although this body is sometimes found within the
cell and not always between the two neighbouring cells. It isa
round body surrounded by a free plasm and staining very deeply
by methylen-blue or Bohmer’s hematoxylin. There is generally
only one but sometimes two. Rarely it has a constriction form-
ing a dumb-bell-shaped rod. When it is present in the cyto-
plasm, it corresponds very well with the centrosome of Platner
(25) who observed it in many Lepidoptera (cf. his figs. 3, 4,
5). After the complete formation of the spindle, it dissolves
and is no more to be seen. Up to the present the function of
this body is quite obscure and requires further investigations.

Besides the ‘“ Verbindungsbriicken’’ we observe in the
cytoplasm an aggregated spot of microsomes which afterwards
changes into the central spindle. It resembles very much in
shape the ‘“‘ Nebenkern”’ of la Valette (35, 36) and originates also
from the cytomicrosomes. Hermann (17), Solger (32), Platner
(25) Ishikawa (22) and others consider the ‘‘ Nebenkern”’ to be
a centrosome with archoplasm. This seems to be the case in
Panulirus japonicus referred to above. But the structure known

146 ON THE SPERMATOGENESIS OF THE SILK-WORM.

as the ‘‘Nebenkern”’ appears not always to be a centrosome
surrounded by an archoplasm, as the ‘‘ Nebenkern”’ of the silk-
worm above described, shows very plainly. The formation of
the ‘‘Nebenkern”’ of the sperm-daughter-cell from the “ Ver-
bindungsfaden”’ of the sperm-mother-cell in the silk-worm con-
firms the opinion of Heymann who describes it in the sperma-
tocyte of Salamandra in the following words (17).: ‘‘ unleugbar
dem Protoplasma entstammend, kehren die Fibrillen der Cen-
tralspindel bei Rekonstruction der Tochterkerne, radienartig
ausstrahlend, wieder in das Protoplasma zurtick.”

ON THE REDUCTION OF CHROMOSOMES.

Since the theoretical consideration of Weismann (44), the
reduction of chromosomes in sperm-mother-cells has been con-
firmed by many eminent authors such as Flemming (8), Hacker
(12), Henking (14, 75, 16), O. Hertwig (18), Ishikawa (20) Platner
(15, 27), and vom Rath (28, 29), in many animals. Especially
by the beautiful researches of O. Hertwig’s “Vergleich der Ei
und Samenbildung bei Nematoden ” not only is this fact proved
but also the similarity of the development in both sperm- and
egg-cells has become clearly known. In general, our researches
on the spermatogenesis of Bombyx mori corresponds so exactly
with the descriptions given by these authors, that it seems almost
superfluous for me to publish them. I have deemed it, however,
worth while to record the results of my own investigations
because there are some points of controversy among the conclu-
sions arrived at by these authors. As the ground of this question

we will quote the researches of O. Hertwig on the Nematodes. .

His summary is:

“1. Die Samenmutterzelle entspricht der Eimutterzelle
oder dem unreifen Ei.

‘‘2. Wéahrend des langer dauernden Ruhezustandes des
ansehnlichen blaschenformigen Kerns der beiden Geschlechts-
producte wird die Kernsubstanz gleich fiir zwei Zelltheilungen,
die sich unmittelbar aufeinander folgen, in eigenartiger Weise
vorbereitet.

HA ~ [1S

ON THE SPERMATOGENESIS OF THE SILK-WORM. 147

“3, Die im Keimblaschen und in dem Samenmutterkern
vorbereitete Menge wirksamer Kernsubstanz ist gleich gross, wie
in jedem andern Kern vor der Theilung. Eine Reduction durch
Ausstossung oder Riickbildung hat nicht stattgefunden.

“4, Wéahrend der zwei sich unmittelbar aufeinander fol-
genden Theilungen findet eine Vermehrung der Kernsubstanz
nicht statt, da das blaschenformige Ruhestaduim des Kerns
ausfallt, und da die im Keimblaschen und Samenmutterkern
vorbereiteten chromatischen Elemente wahrend der zwei Theil-
processe weder an Masse zunehmen, noch sich der Lange nach
spalten. Die aus dem zweiten Theilungsact hervorgehenden
Endproducte enthalten daher in Folge der zweimal eingetretenen
Halbirung nur die Halfte der Kernmasse, welche ein gewohn-
licher Kern nach der einfachen Theilung besitzt.

*¢5. Die Anzahl derim Samenmutterkern und Keimblaschen
vorbreiteten chromatischen Elemente ist bei Ascaris ebenso gross,
wie bei einem gewohnlichen Kerne in der Mitte des Theilungs-
processes, also die doppelte wie sie ein Kern in der Vorphase
der Theilung zeigt. Der morphologische Werth dieser Elemente
scheint aber ein anderer zu sein, in Folge einer von normalen
Verlauf abweichenden Entstehung. Wa&ahrend normaler Weise
acht Tochterchromosomen durch einfache Langsspaltung von
vier Faden entstehen, scheinen sie hier durch doppelte Langs-
spaltung von nur zwei Faden gebildet worden zu sein. Diese
zwei Faden enthalten aber dieselbe Substanzmenge, wie vier
durch Quertheilung am Anfang der Karyokinese gebildete
Faden.

“6. Da die Eimutterzelle und die Samenmutterzelle diesel-
ben Kerntheilungsprocesse mit allen ihren von der Norm abwei-
chenden Eigenthiimlichkeiten in genau der gleichen Weise
durchmachen, miissen die Theilproducte auch denselben mor-
phologischen Werth besitzen.

““a) Den beiden Samenmutterzellen entsprechen Ei-und
erster Richtuugskorper.

‘*b) Den vier Samenenkelzellen (Samenkorpern) sind das reife
Ei, der zweite Richtungskérper und die aus Theilung des ersten
Richtungsk6rpers entstehenden zwei Kiigelchen zu vergleichen.

“‘c) Die Richtungskérper haben daher den morphologischen
Werth rudimentarer Eizellen.”

148 ON THE SPERMATOGENESIS OF THE SILK-WORM.

The investigations of vom Rath (28) on the spermatogenesis
of Gryllotalpa and of Hacker (12) on the egg-formation of
Cyclops and Canthocamptus, correspond very well with Hert-
wig’s results, and Wetsmann summarises the facts in the follow-
ing words: ‘In all those species which have been investigated
for this purpose, the germ-cells are formed by the mother-
cell undergoing two consecutive divisions, each of which results
in a halving of the number of idants, one half passing into
the one daughter-cell, and the other half into the other. In
the second division this would lead to the presence of only a
quarter of the original number of idants, if the number in the
mother-cell were not doubled by each idant becoming split into
two before the first division takes place. Thus there is first
a doubling, and then a halving, of the number of idants. I
consider this remarkable and apparently useless process of the
doubling and two subsequent halvings of the idants as a method
of still further increasing the number of possible combinations
of idants in the germ-cell of one and the same individual, and
have given reasons for this opinion in the above named essay.”

Prof. Ishikawa’s results on the reproductive elements of
Diaptomus sp. also correspond with the results of the above
named authors except the mode of division, in which he describes
the transverse constriction of the chromosomes in the first divi-
sion of the sperm-mother-cells giving rise to dumb-bell-shaped
bodies.

Henking’s researches on the genital elements of many insects
(16, 15) are different from the results of all the above named
authors, in the time of the reduction of chromosomes. According
to this author the reduction of chromosomes takes place in the
first division of the spermatocytes or sperm-mother-cells and
no doubling of chromosomes takes place in the growing stage
of these cells, while their second division is to be looked upon
as the ‘‘ Equationstheilung.”

In Bombyx mori, as stated above, twently-eight chromo-
somes are found in the primary germ-cells. In the growing
stage I could not detect any longitudinal splitting or the doubl-
ing of the number of chromosomes as vom Rath gives it in the
sperm-mother-cells of Gryllotalpa (28). Here I am more in-
clined to agree with Henking (15) who denies its occurrence.

ON THE SPERMATOGENESIS OF THE SILK-WORM. IA49

In the first division of the sperm-mother-cells (‘‘ Sperma-
tocyten 1. Ordn.”), the twenty-eight chromosomes in the equa-
torial plane of the spindle divide transversely, as stated above,
and produce two rows. Both Henking and vom Rath affirm
also the existence of two rows of chromosomes in the equator
of the spindle of this stage. But this, according to vom Rath
(28), is due to the doubling of the chromosomes in the growing
stage of the sperm-mother-cells, while Henking (15) believes it
to be formed by the double arrangement of the chromosomes,
already existing, so that according to this latter author, the
reduction of the number of chromosomes takes place by
this division, which according to all other authors occurs by
the second “),

The reduction of the number of chromosomes in the genital
cells of plants has been worked over, so far as I know, only by
Guignard (10). The mode of the reduction which he observed of
Lilium martagon, however, differs entirely from that of animals.
According to this investigator the reduction occurs without a
nuclear division in a stage of the ‘‘cellules mére définisives” (pro-
bably corresponding to the sperm-mother-cells of animals). My
investigation on the formation of the pollens of Lilium tigrinum
and Allium fistulosum is not in accordance with this, but shows
much similarity with that of animals. This I trust will be
soon published.

SUMMARY.

1. A large cell in the blind end of the follicle which is de-
signated by the name of “‘ Keimzelle”’ by Verson, is not a genital
cell but a supporting cell which connects the genital cells with
the follicular wall. It always has a large nucleus with finely
granular chromosomes collected here and there and staining
very deeply with hzmatoxylin, safranin, carmine, and anilin
colours.

(1) According to the investigations of Moore (24), the sperm-mother-cells of the
rat contain sixteen chromosomes, which are reduced into eight, and “the reduction
would appear more comparable to the type described by Guignard during the forma-
tion of vegerable pollen, than, to that pointed out by Hertwig in the spermatocytes
of Ascaris.” This however appears to me very improvable.

150 ON THE SPERMATOGENESIS OF THE SILK-WORM.

2. The primary sperm-cell resembles very much the primary
egg-cell, and is conical in shape and contains a nucleus in its
broad end. Its nucleolus always consists of an aggregation of
chromatin granules, whose number varies from two to four or
more. It divides many times in the usual karyokinetic manner,
and the cell-body gradually decreases in size until it becomes
about two-thirds or less of the original cell. It is then called
the sperm-mother-cell and passes to the growing stage.

3- In the early part of the growing stage, the sperm-mother-
cell has an ordinary resting nucleus with the chromatin granules
scattered in it. It gradually becomes larger, and with this
the change of nucleus takes place. The chromatin granules
first gather themselves to one side of the nucleus and form an
irregular mass from which fine achromatic fibres radiate. This
chromatin mass.dissolves and a fine skein stage with a nucleolus
is formed. The chromatin granules of this skein again dissolve
into many isolated granules and the nucleus again assumes a
granular structure. There are now two nucleoli in each nucleus.
These nucleoli pass out into the cytoplasm one after the other.
An achromatic spindle is also seen at this stage, formed from
cytomicrosomes. This spindle consists of two parts, the central
and the polar, which latter seems to be derived from the nucleus
together with the centrosomes. The granular chromosomes,
now again collect themselves and form ‘ring-like structures.
Each of these rings dissolves again into four chromosomes and
forms “ Kernplatte.”

4. The sperm-mother-cells contain at first twenty-eight
chromosomes. In the first division each of these divides trans-
versely to its long axis and forms two chromosomes, of which
one goes to one pole and the other to the opposite pole. The
daughter-cells thus contain each twenty-eight chromosomes as
the mother-cell.

The nuclei of these daughter-cells form no resting stage,
and directly pass over to the second division in which half the
number of chromosomes goes to one cell and the other half to
the other. The grand-daughter-cells thus formed contain there-
fore only fourteen chromosomes, which is half the number of

the mother-cell.
_ 5. After this division, the fourteen chromosomes nlees at

ON THE SPERMATOGENESIS OF THE SILK-WORM. ape

the periphery of the nucleus and finally form a compact, deeply
stainable mass with pointed end and vacuole. This chromatin
mass gradually elongates and forms a spindle-shaped head of a
spermatozoon. From the remnants of the “‘ Verbindungsfaden,”’
a ‘‘Nebenkern” is formed, which elongates and enters into the
tail part. A mitosome is also seen, arising from the cytomicro-
somes.

End of December, 1893.

10.

21Ie

22.

LIST OF WORKS REFERRED TO.

Van Beneden, Ed.—Recherches sur la maturation de l’oeuf et la fécondation.
Arch. d. Biologie. T. IV. 1883.

Boveri, Th.—Zellen-Studien. Heft 2. Jena. 1888.

Brauer, A.—Zur Kenntniss der Herkunft des Centrosomas. Biol. Centralbl.
Bd. XIII. 1893.

Brauer, A.—Zur Kenntniss der Spermatogenese von Ascaris megalocephala.
Arch. f. mikr. Anatomie. Bd. XXXXII. 1893.

Cornalia, E.—Monographia del bombice del gelso. 1856.

Cholodkowsky, N.—Zur Kenntniss der mannlichen Geschlechtsorgane der Dip-
teren. Zool. Anzeiger. 1892.

Flemming, W.—Neue Beitrage zur Kenntniss der Zelle. Arch. f. mikr. Anato-
mie. Bd. XXIX. 1887.

Flemming, W.—Neue Beitrage zur Kenntniss der Zelle. II. Arch. f. mikr.
Anatomie. Bd. XXXVII. 1891.

Flemming, W.—Weitere Beobachtungen uber die Entwicklung der Spermato-
somen bei Salamandra maculosa. Arch. f. mikr. Anatomie. Bd. XXXI.

Guignard, L.—Nouvelles études sur la fécondation. Ann. Sc. Natur., Bota-
nique. T. XIV. 18g1.

Haberlandt, F.—Der Seidenspinner des Maulbeerbaumes. Wien. 1871.

Haecker, V.—Die Eibildung bei Cyclops und Canthocamptus. Zoolog. Jahr-
bicher. Bd. V. Abth. f. Morph. r8q1.

Heidenhain, M.—Uber Kern und Protoplasma. Festschr. f. Prof. von Kolliker.
1892.

Henking, H.—Uber Reduktionstheilung der Chromosomen in den Samenzellen
von Insekten. Internat. Monatsschr. f. Anat. u. Physiol. Bd. VII. 1890.
Henking, H.—Untersuchungen uber die ersten Entwicklungsvorgange in den
Eiern der Insekten. II. Uber Spermatogenese und deren Beziehung zur
Eientwicklung bei Pyrrhocoris apterus, L. Zeitschr. f. wiss. Zoologie. Bd.

LI. 18or.

Henking, H.—The same. III. Specielles und Allgemeines. Zeitschr. f. wiss.
Zoologie. Bd. LIV. 1893.

Hermann, F.—Beitrage zur Lehre von der Entstehung der karyokinetischen
Spindel. Arch. f. mikr. Anat. Bd. XXXVII.

Hertwig, O.—Vergleich der Ei- und Samenbildung bei Nematoden. Arch. f.
mikr. Anat. Bd. XXXVI. 1890.

Hertwig, O.—Die Zelle und die Gewebe. Jena. 1893.

Ishikawa, C.—Studies of reproductive elements. I. Spermatogenesis, ovoge-
nesis and fertilization in Diaptomus. Jour. of the Coll. of science. Imp.
Univ. Japan. Vol. V. 1891. '

Ishikawa, C.—The same. II. Noctiluca miliaris, Sur.; its division and spore-
formation. Jour. of the Coll. of Science. Vol. VI. 1894.

Ishikawa, C.—Uber die Kerntheilung bei Noctiluca miliaris. Bericht. d.
naturf, Gesellschaft z. Freiburg. Bd. VIII. 1893.

23.

24.
25.

26.

27.

28.

29.

LIST OF WORKS REFERRED TO. 153

Karsten, G.—Uber Beziehungen der Nucleolen zu den Centrosomen bei Psilo-
tum triquetrum. Bericht d. Deuts. Bot. Gesellschaft. 1893.

Moore, J. E. S.—Mammalian Spermatogenesis. Anat. Anzeiger. 1893.

Platner, G.—Beitrage zur Kenntniss der Zelle und ihrer Theilungserschei-
nungen. Arch. f. mikr. Anat. Bd. XXXIIT. 1880,

Platner, G.—Uber die Entstehung des Nebenkerns und seine Beziehung zur
Kerntheilung. Arch. f. mikr. Anat. Bd. XXVI. 1886.

Platner, G.—Die Karyokinese bei den Lepidopteren als Grundlage fir eine
Theorie der Zelltheilung. Inter. Monatschr. f. Anat. u. Histol. Bd. III.
1886.

Vom Rath, O.—Zur Kenntniss der Spermatogenese von Gryllotalpa vulgaris
Latr.. Arch. f. mikr. Anat. Bd. XL. 1892.

Vom Rath, O.—Beitrage zur Kenntniss der Spermatogenese von Salamandra
maculosa. I. Theil. Die Reduktionsfrage. I]. Theil. Die Bedeutung der
Amitose in Sexualzellen und ihr Vorkommen im Genitalapparat von Sala-
mandra maculosa. Zeitschr. f. wiss. Zoologie. Bd. LVII. 1893.

Rabl, C.—Uber Zelltheilung. Morphol. Jahrbuch. Bd. X. 1885.

Schewiakoff, W.—Uber die Karyokinetische Kerntheilung der Euglypha alveo-
lata. Morphol. Jahrb. Bd. XIII. 1888.

Solger, B.—Zelle und Zellkern. Thiermed. Vortrage. Bd. IIT. 1892.

Strasburger, Ed.—Histologische Beitrage. Heft. 1. Uber Kern- und Zelltheilung
im Pflanzenreiche etc. Jena. 1888.

Strasburger, Ed.—The same. Heft. IV. Schwarmsporen, Gameten, pflanzliche
Spermatozoiden, und, das Wesen der Befruchtung. Jena. 1892.

St. George, von la Valette.—Spermatologische Beitrage. Heft II. Arch. f.
mikr. Anat. Bd. XXVII. 1886.

St. George, von la Valette.—Zelltheilung und Samenbildung bei Forficula
auricularia. Festschr. f. von Kélliker. 1887.

Tichomiroff, A.A—Entwicklungsgeschichte von Bombyx mori L. im Ei (Russ.)
Moskau. 1882.

Verson, E.—La spermatogenesi nel Bombyx mori, L. Padova. 1889.

Verson, E.—Zur Spermatogenesis. Zoolog. Anzeiger. Jahrg. XII. 1889.

Waldeyer, W.—Uber Karyokinese und ihre Beziehungen zu den Befruchtungs-
vorgangen. Arch. f. mikr. Anat. Bd. XXXII. 1888.

Watase, S.—Karyokinesis and the cleavage of the ovum. Johns Hopkins
University Circulars. Vol. IX. 18go.

Watase, SimHomology of Centrosome.* Jour. of Molphol. Vol. VIII. 1893.

Von Wasielewski.—Die Keimzone in den Genitalschlauchen von Ascaris mega-
locephala. Arch. f. mikr. Anat. Bd. XXXXI. 1892.

Weismann, A.—Essays upon heredity and kindred biological problems. Vol.
I. Oxfold. 1891.

Weismann, A.—The Germ-plasm. (English translation of ‘‘Keimplasma”’).
London. 1893.

Ziegler, H. E., und vom Rath, O.—Die amitotische Kerntheilung bei den
Arthropoden. Biolog. Centralblatt. Bd. XI. 1891.

EXPLANATION OF PLATES, III—IV.

All figures are drawn with Abbe’s camera. lucida as well as drawing prism.
Objectives and oculars used, are mentioned with explantions of figures.

Bigs rs

ee 2
Fig, 3
Fig. 4
Fig. 5.
Fig. 6
Fig. 7
Fig. 8.
Fig. 9.
Fig. ro.
Fig. 11.

Surface view of male sexual organ of larva two days old. Killed with
picro-acetic acid and stained with alcohol carmine. (¢) testis, (d) dorsal
vessel. (Zeiss, Obj. D, Oc. I).

Surface view of female sexual organ of larva two days old. Similarly
treated as above. (0) ovary, (d) dorsal vessel. (Zeiss, Obj. D, Oc. I).
Terminal portion of vas deferens of larva in fifth stage. (m) muscle, (e)
uterus-shaped process of ventral median side of twelfth segment. Treated
as above. (Leitz, Obj. 1, Oc. 4).

Longitudinal section of testis of embryo, about two days before hatching.
It will be seen that here testis consists of single cavity with round genital
cells and ‘‘ Hodenzwischenkérperchen”’ (X) scattered in it. Killed with
1°/, formic acid and stained with borax carmine. (Zeiss, Obj. J, Oc. 3).
Longitudinal section of testis of larva just hatched. (/) first three de-
pressions on follicular wall; (i) second four invaginations. Killed with
Flemming’s stronger solution and stained with Hermann’s triple staining.
(Leitz, homog. imm. 1/12, Oc. 3).

Longitudinal section of testicular follicle, of larva three days old, with
invaginated follicular wall and supporting cell (v) (Verson’s cell). Treated
as above. (Leitz, homog. imm. !/;2, Oc. 2).

Longitudinal section of testicular follicle of larva, one day after fourth
moult: left side of figure represents formative zone and right side is
place where vas deferens takes its origin. (v) Verson’s cell; (h) ‘‘ Hoden-
zwischenkorperchen.” Treated as above. (Zeiss, Obj. BB, Oc. 4).
Blind end of testicular follicle, showing protoplasmic strand (f) connecting
Verson’s cell with follicular wall. Killed with Rabl’s platinum chloride
and stained with Bohmer’s hematoxylin. (Zeiss, Obj. D, Oc. 4).

Verson’s cell, with small degenerating cells surrounding it; taken from
larva within coccoon. Killed with Flemming’s stronger solution and
stained with acid fuchsin. (Leitz, Obj. 7, Oc. 4).

Small portion of testicular follicle of larva in third stage. Lower part
of figure represents blind end of follicle. Wall seen at right hand
side is follicular wall. (v) Verson’s cell; (a) free area at blind end of
follicle in which Verson’s cell is normally to be seen. Treated with
Flemming’s stronger solution and Hermann’s anilin-water saffranin.
(Leitz, homog. imm. 1/12, Oc. 3).

Verson’s cell (v) with surrounding protoplasm and primary germ-cells
attached to it. Killed with picro-acetic acid and stained with Delafield’s
hematoxylin. (Leitz, homog. imm. 2/12, Oc. 3).

NI

EXPLANATION OF PLATES, III-—IV. 155

Fig. 12, Verson’s cell (v) with surrounding small degenerating cells, taken from
testis of imago. Killed with Flemming’s stronger solution and stained
with rubin. (Leitz, Obj. 7, Oc. 4).

Fig. 13. Verson’s cell (v) with degenerating cells, taken from testis of imago after
copulation. Treated as above. (Leitz, obj. 7, Oc. 4).

Fig. 14. Longitudinal section of terminal portion of ovarian tube, taken from larva
in third stage. (v) Verson’s cell. Treated with Flemming’s stronger
solution and stained with Flemming’s saffranin solution. (Leitz, homog.
imm. 1/12, Oc. 3).

Fig. 15. Testicular follicle of Papilio xuthus with Verson’s cell (v). Treated as
above. (Leitz, Obj. 7, Oc. 4).

Fig. 16. Testicular follicle of wild silk worm with Verson’s cell (vj. Treated as
above. (Leitz, Obj. 7, Oc. 4).

PEATE IV.

All specimens killed with Flemming’s strong solution, except those shown in

figs 63, 64, 82, 81 and 86.

Fig. 17. Resting nuclei of primary germ-cells. (7) nucleolus, consisting of chro-
matin granules. Treated with Hermann’s triple staining. (Zeiss, homog.
imm, 1/12. Oc 4.)

Fig. 18. Two nuclei of same stage with greater portion of cell-bodies. Treated as
above. (Zeiss, homog. imm, 1/12, Oc. 5.)

Fig. 19. Two primary germ-cells at beginning of skein stage. Treated as above.
(Zeiss, homog. imm. 1/12, Oc. 4).

Fig. 20. Five primary germ-cells, with their Nuclei at stage of segmented skein.
In Nucleus represented at right hand side of figure chromosomes are
much shortened. Treated as above. (Zeiss, homog. imm. 2/12, Oc. 4).

Fig. 21. Stage of division of primary germ-cells. In left lower nucleus, seen from
polar view, twenty-eight chromosomes can be distinctly counted. Treated
as above. (Zeiss, homog. imm. 1/12, Oc. 5).

Fig. 22. Primary germ-cells of more advanced stage of division, ‘Twenty-six, or
twenty-seven chromosomes are to be counted_in polar views. Treated as
above. (Zeiss, homog. imm., 1/12, Oc. 4).

Figs. 23-45. Successive stages of metamorphosis of sperm-mother-cells in growing
stage. Figures 39, 40, 41, 42, and 43 are stained with Bohmer’s he-
matoxylin, and all others with Hermann’s triple dyes,

Fig. 23. Resting nuclei. These are very small in consequence of repeated divi-
sions in last stage. (Zeiss, homog. imm. 1/12, Oc. 4).

Fig. 24. Resting nuclei at more advanced. (Zeiss, homog. imm. 2/12, Oc. 4).

Figs. 25 and 26. Nuclei, before skein stage. Chromatin granules are collected
irregularly at one side of nucleus. Nucleolus consists of small chromatic
bodies. (Zeiss, homog. imm, 1/12, Oc. 4).

Fig. 27. Skein stage., (Zeiss, homog. imm. 1/12, Oc. 4).

Figs. 28, 28’. Granulation of chromosomes. (Zeiss, homog. imm., 2/12, Oc. 4).

Fig. 29. More advanced stage as above. Here chromosomes are completely dest-
royed. (ditto).

156 EXPLANATION OF PLATES, III—IV.

Fig. 30. Chromatin bodies represented in last figure are now collected at centre
of nucleus. Single nucleolus is seen in centre of these chromatin bodies.
(ditto).

Fig. 3z. Nucleus in which chromatin granules again separate from one another
and show appearance of normal resting nucleus. Large nucleolus consist-

ing of small chromatin granules is seen. (ditto).

Figs. 32-43. Sperm-mother-cells nearly in same stage of development. (v) ‘ Ver-
bindungsbriicken’’; (c) extruded nucleolus; (e) nucleus of cyste. In fig.
40 collection of cytomicrosomes (6) is shown. (Same magnification as
above.)

Fig. 44. More advanced stage. Chromosomes show ring-like form. (Zeiss, homog.
imm. 1/12, Oc. 4).

Fig. 45. Similar stage as above. Each ring broken up into four chromosomes.
(Same magnification as above.)

Figs. 46-53. Various stages of first division of sperm-mother-cells. Figs. 46-48 re-
present early stages of division, showing attraction-spheres, oblique
spindle and ‘‘ Verbindungsbriicken”’ (v); and in fig. 49 polar and central
spindles are seen. Stained with Béhmer’s hematoxylin. (Zeiss, 1/12,
Oc. 4). Figs. 50, 50’, 51, 52, Spindle stage. Fig. 53 diaster stage of
the same. Treated with Hermann’s triple staining. (Zeiss, 1/12, Oc. 5
except fig. 50’ which is drawn by Zeiss. t/12, Oc. 5.)

Figs. 54-60. Second division of sperm-mother-cells, immediately following last divi-
sion. Figs. 54 and 55, spindle stage. (Zeiss, homoz. imm. 1/12, Oc. 5).
Fig. 56, diaster stage of same. (Zeiss, homog. imm. 1/rz, Oc. 4). Fig.
57 shows polar view of diaster stage. (Zeiss, Obj. 1/12, Oc. 5). Fig.
58, side viw of same. (Magnif. as above). Fig. 59, little more advanc-
ed stage. Central and polar spindles have separated from each other.
(magnified as above). Fig. 60, more advanced stage of same, showing
two cell-plates; (&) nucleus. (Zeiss, Obj. 1/12, Oc. 4), Those shown
in figs. 54, 55, 50 stained with Hermann’s triple staining while rest are
treated with Bohmer’s hematoxylin.

Figs. 61-83. Various stages of metamorphosis of sperm-daughter-cells. Figs. 61,
62, a, b show sperm-daughter-cells with remnant of spindle fibre and
collected mirosomes (m). (Zeiss, Obj. t/r2, Oc. 4. and 5). Stained with
Bohmer’s hematoxylin. Fig. 62c, d represent more advanced stage of
the same, showing ‘‘Nebenkern”’ (n). Treated and magnified as above.
In figs. 63 and 64, cell-body is little elongated to form tail of sperma-
tozoon; (k) nucleus (7) ‘‘ Nebenkern” (m) mitosome are shown. Acetic
acid methyl green preparation and magnified by Zeiss, Obj. J, Oc. 4,
Figs. 65-78, 74-76, 78-81 show head and ‘‘ Nebenkern,”” of spermatozoa
showing successive stages of metamorphosis. (K) nucleus, (7) ‘‘ Neben-
kern,” (m) mitosome. Béhmer’s hematoxylin except fig. 72 which is
stained with acid fuchsin and methylen blue. Magnified by Zeiss,
homog. imm. !/rz, Oc. 5. except figs. 67, 69, 72, 74, 75 which are drawn
with Zeiss, homog. imm. 2/rz, Oc. 4. Fig. 81 shows head of nearly
mature spermatozoon, killed with picro-acetic acid and stained with
picro-carmine. Fig. 82 represents cyste containing nearly matured
spermatozoa. At right end of figure large supporting cell is. shown.
Treated as above. Zeiss, homog. imm. 1/12, Oc. 3. Fig. 83 shows

EXPLANATION OF PLATES, III—IV. 157

entire spermatozoon in same stage as fig. 74. (homog. imm. 1/12, Oc. 4.)
Fig. 69, b. Cross section of tail of spermatozoon at same stage as fig. 69 a, passing

through ‘*Nebenkern.”’ Stained with Hermann’s triple staining and
drawn by Zeiss, homog. imm. 1/12, Oc. 5.

Fig. 73. Similar section of a spermatozoon at the same stage as that shown by
fig. 70. Treated and magnified as above.

Fig. 77. Similar section through tail of spermatozoon at same stage as that

shown by fig. 83. Treated and magnified as above

Figs. 84-86. Various stages of ‘‘ Hodenzwischenkérperchen.” Figs. 84 and 85
stained with Hermann’s triple staining; fig. 86 with Bohmer’s hema-
toxylin. Fig. 84 and 86, Zeiss, homog. imm, 1/12, Oc. 4; fig. 86 Zeiss,
homog. imm. 1/12, Oc. 5.

Figs. 87-90. Sperm-mother-cells of Panulirus japonicus, showing resting nucleus
with centrosome and archoplasm. Killed with Flemming’s stronger solu-
tion and stained with anilin-water saffranin. (Zeiss, homog. imm. 2/12
Oc. 3).

Bull, Agric. Coll. Vol. I. Pl. II.

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The Energy of the Living Protoplasm,

BY

Dr. Osear Loew, Professor of Agricultural Chemistry.

CHAPTER VI.
The Chemical Activity of Living Cells.

The great chemical activity of living cells astonishes the
pondering mind. The synthesis of carbohydrates, the transforma-
tion of starch into cellulose and fat, the production of the highly
complicated proteids are executed with admirable readiness by
plant-cells. Animal protoplasm also—although in extent and
power of chemical synthesis far excelled by vegetal protoplasm—
is capable of many highly interesting chemical operations, such
as the construction of living protoplasm out of inert albuminous
matter, the formation of hemoglobin, mucin, elastin, keratin,
and glutin from the proteids of the food, the production of
enzymes, or the transformation of sugar into fat. And not less
remarkable than the synthetical work is the energetic oxidation
exhibited by the respiration process.)

The character of the chemical work in plant-cells is many-
sided: polymerisation, condensation, esterification, formation of
amido-compounds, generation of the cyclic constitution, reduc-
tions and oxidations are included in it. We find acids and
bases, aldehydes and ketones, esters, alcohols, and sulphides in
the vegetal kingdom; but neither aldoximes, nor ketoximes,‘”
neither sulpho-acids, nor nitro-, nor azo-compounds. ‘Thus, the
chemical activity, varied as it is, seems, nevertheless, not to
leave certain channels.°) A number of chemical operations

(1) See the following chapter.

(2) No derivative of hydroxylamine has as yet been discovered in plant-cells.

(3) It is a notable fact, for which hitherto explanation has not been given, that
especially such benzene derivatives as have a lateral chain of three carbon atoms
frequently occur in plants, some in form of glucosides. I will mention: hesperidic
acid, caffeic acid, melilotic acid, cumarin, umbelliferone, daphnetin, coniferyl alco-
hol, eugenol, anethol, safrol, cinnamic alcohol and aldehyde, cinnamic acid, thymol,
scopoletin, asarone, syringin, zsculetin, absinthol, camphor, the terpenes. Also,
tyrosin and phenylamidopropionic acid, as decomposition-products of proteids, have
to be mentioned (See Ch. V).

| ae

160 THE ENERGY OF THE LIVING PROTOPLASM.

can be carried out by every plant-cell, others only by specifically
endowed ones, and, while certain compounds are found in all
cells, such as carbohydrates, fats, and proteids, others are not of
universal occurrence, though still very frequent, as tartrates,
succinates, glucosides, resins, tannin.“) Others, again, are very
rare, being either restricted to one family, as quinine or strych-
nine, or to a few families, as caffein.‘®

In scrutinising the chemical activity of a plant-cell, we have
at first to pay attention to the differentiation of the plasmic
contents and the division of chemical labour. The cortical layer
of the cytoplasm has not the same function as the inner layer
or tonoplast; the former produces the cellulose-wall from starch
or sugar, while the latter restrains noxious substances accumula-
ted in the vacuole from returning into the protoplasm. ‘The
Ieukoplasts form starch from sugar, thereby preventing a higher
concentration of the sugar-solution which would to some
extent check plasmic activity. The chloroplasts (“ chlorophyll-
granules”), again, with the aid of ethereal oscillations of a deter-
mined wave-length, transform carbonic acid into sugar.%) The
nucleus, finally, is charged with the duty of producing the necessary
enzymes; diastase is required to bring starch into solution for
purposes of transportation and transformation, and proteolytic
enzymes to utilise aleurone-grains, during the germination process.

(1) For the chemical relations and full descriptions of tannins see Henry
Trinble, The Tannins, Philadelphia, 1894.

(2) Cf. A. Husemann and A. Hilger, Die Pflanzenstoffe; also Ed. Schdar,
Schweizer Wochenschrift fiir Pharmacie, 27, 197. Certain of the accessory com-
pounds may still serve some biological purpose as that of attracting insects for fecun-
dation, or of protection against fungi and animal parasites, while others are useless
by-products and mere excretions.

(3) Carbonic acid is often called a food for plants, although it is only the material
from which plant food (glucose, etc.) is prepared in the chlorophyl-bodies, a logical
distinction pointed out by F. Stohmann (Z. Biol, 31, 365). In the assimilation of
carbonic acid it is generally assumed that formic aldehyde is the first product. This
yields, however, upon condensation in solution, as I have shown, not dextrose but
other sugars; moreover, it is poisonous. Therefore, I have added the hypothesis
that the formic aldehyde first formed combines with certain hydroxyl-groups in the
protoplasm of the chloroplasts, the amido-groups being protected. The condensation
taking place afterwards must thus lead always to one and the same configuration of
the resulting sugar, since the molecules of formic aldehyde have lost their freedom of
motion. Cf. O. Loew, Ber. D. Chem. Ges. 1889, 484; also, ibid., 473 and 474.

THE ENERGY OF THE LIVING PROTOPLASM. 161

Plant-cells deprived of their nucleus are incapable of trans-
forming starch into cellulose (K/ebs). They evidently cannot bring
on a sufficient saccharification of the starch granules, to provide
the cytoplasm with the necessary amount of sugar. For the
lowest animal forms, an analogous case was proved by B. Hofer ;
amoebe can no longer digest albuminous particles when deprived
of their nucleus. ‘The importance of the nucleus for the secretion
processes of animal organisms was also observed by Verworn,
Balbiam, and Korschelt. The latter found, for example, consi-
derable changes of form of the nuclei in insects during the
secretion of chitin.

The vegetal nucleus seems to be also the manufacturer of
the proteids, as first suggested by Strassburger and Schinitz ;
at least protein-crystalloids are frequently found in the nuclei
of plant-cells, especially in the orders, Oleacee, Scrophularinea,
Bignoniacee, and in the Pteridophytes (Zimmermann). For,
it does not seem probable that this protein has its source in
the cytoplasm and is then transported into the nucleus for
crystallisation. Again, in germinating seeds, where protein
formation from fragments of decomposed reserve-proteids proceeds
with considerable readiness, the nuclei (and also the nuc-
leoli) increase in size.) “The most important function of the
nucleus, however, is connected with the division and multiplica-
tion of cells and with sexual propagation in plants and animals.

(t) Peters, Botan. Centralbl. 48,181. The nuclei are also often relatively large
in the epidermis-cells of leaves, which are especially adapted to store up larger
quantities of active albumen (Bokorny, Pfliig. Arch 55, 142). Also in the cells of
glands relatively large nuclei are often present.

It is also a remarkable observation that plant-cells deprived of their nucleus do
not produce any more starch by assimilation. Only certain algze, containing
pyrenoids (Zygnemacez) have thus far been observed to be exceptions in regard
to starch-production under this condition (Klebs). Simpler organisms, like amcebze
and alge, are especially well suited for experiments with isolated cytoplasm. Klebs
has, by plasmolysis, Gerasimoff by low temperature during the process of karyokinesis,
obtained living parts of alge-cells, or living cells without a nucleus. The life of such
cytoplasm, however, did not last longer than about six weeks. On the other hand the
nuclei of radiolaria can remain alive, if deprived of the cytoplasm, only 10-15 hours,
Nucleus and cytoplasm influence each other by certain of their products (Verworn
Pfliig, Arch. 51, 113).

162 THE ENERGY OF THE LIVING PROTOPLASM.

As the main constituent of the nucleus is nuclein' and as this
is essentially a combination of an albuminous substance with
phosphoric acid’) the importance of the latter becomes at once
conspicuous. The important réle of the nuclein in the division
of the nucleus has been demonstrated by staining methods, but
it may also be inferred from observations on algz cultivated in
liquids devoid of phosphates. If we place some filaments of
Spirogyra majuscula in about a liter of distilled water, to which
has been added 0.2 p. mille calcium nitrate and 0.02 p. mille
ammonium sulphate, and expose the culture to diffused daylight,
assimilation as well as protein formation can proceed to a certain
extent®’ and even growth of the cells to a considerable size
will take place, but 20 multiplication of cellsis noticed. Gradually,
however, the threads assume a yellowish tinge and commence to
suffer. If after about six weeks we divide the filaments, together
with the solution, into two portions, adding to one 0.02 p. mille
protosulphate of iron, to the other, besides this, 0.08 p. mille
disodium phosphate, we can observe in a few days, in the latter
case, not only the restoration of the brilliant green colour but also
the process of karyokinesis, in almost every cell, and this too, in the
daytime. In the former case, however, where the iron salt
alone was added, none of these phenomena was observed,
showing, further, that for the production of a normal chlorophyll,
phosphoric acid is just as important as iron salts.

The nucleus plays also a certain part in the organisation of
the cytoplasm. In cases of mutilation, only such cells can be
regenerated as still contain the nucleus (Nussbaum, Gruber).
That the growth of vegetal cells bears a certain relation to the

(1) According to E. Zaccharias, besides nuclein, plastin also occurs in the nucleus
and chloroplasts. Plastin appears to be mainly distinguished from nuclein by its
greater resistance to acids and alkalies.

(2) Liebermann showed that nuclein contains metaphosphoric acid, and Kossel,
that bases of the xanthin series and nucleic acid are contained in it. My own
observations make it highly probable that the nuclein in the living nucleus is present
as a lime compound, that is to say, in chlorophyll-bearing plants. With fungi,
however, it is different, lime not being at all required by them. Cf. O Loew, On the
functions of calcium and magnesium salts in plants, Flora, 1892, 368; Landw,
Versuch-Stat. 41, 467.

(3) Traces of potassium salts are probably always stored up in the plant-cells.

THE ENERGY OF THE LIVING PROTOPLASM. 163

nucleus was observed by KAlebs and Haberlandt,") so that
it is probable that the nucleus prepares the proteids suitable
for the purpose of organisation. The vegetal nucleus performs
this by far-reaching synthesis, the animal nucleus by transform-
ing the resorbed peptone.

Clearly, the nucleus of the living cell cannot be identical
with that of the dead, and the nuclein known to chemists, ex-
tracted with alkalies and precipitated with acids, is a relatively
stable compound, which would be entirely incapable of serving
the physiological phenomena of karyokinesis, which are made
possible only by a highly labile state of the proteid composing
the nucleus.)

Division of labour, still of restricted compass in a single
plant-cell, keeps growing in importance and extent with the
development of multicellular organisms, plants as well as
animals. Some cells serve mechanical devices, others do

(1) Biol. Centralbl. 8, 133.

(2) I consider the process of organisation as a sort of polymerisation, in which
only molecules of equal configuration can participate, and in which isomeric and stereo-
isomeric molecules would be hurtful, the mutual reaction of their labile groups being
facilitated (Cf. O. Loew, Natural System of Poisonous Actions, Ch. V. On the
poisonous proteids, 82). This, again, would imply a specific tectonic for the nuclei
of different species as the necessary condition for yielding one and the same active
albumen for the organisation of each cytoplasm. This view could also furnish a
plausible explanation of certain observations in regard to the propagation of the
species. A specific configuration of the nuclein will naturally lead to a specific
tectonic of the nucleus, i.c., the invisible anatomical structure will be in close connec-
tion with the configuration of the proteid. Such considerations may finally also
throw some light upon the fact, that the fibrins, the oxyhemoglobins, the alexines of
different species are by no means identical. The possible stereo-isomers of a proteid
reach evidently to an immense number, even if we start from one and the same
active peptone. [Stereo-isomerism conceives of compounds as containing the same
elements in the same proportion and arranged in the same groups, and yet differing
in properties, because of a different arrangement in space of the constituent groups].

(3) Various observations prove that the nucleus is even of much greater lability
than the cytoplasm. The problem of karyokinesis appears still more complicated now
that the centrosomes have been observed, initiating the division.

Differences have been observed between the nuclei of nerves, glands, and
muscles ; further, between those of the male and female sexual cells, the former being
richer in nuclein (E. Zaccharias). Still greater must be the differences between the

nuclei of the germ-cells and those of the somatic cells, especially in animals.

164 THE ENERGY OF THE LIVING PROTOPLASM.

specific chemical work, and others again are adapted to functions
of propagation, while in the animal kingdom special organs for
visible motion and for sensation are developed. ‘The ‘‘ chemical
factories in the service of an organism,” the glands, of simple
structure in plants, but forming complicated organs in the
higher animals, betray again a far-reaching differentiation.
They may secrete enzymes, carbohydrates, different acids, fat,
or wax. Products secreted by vegetal cells alone are terpenes
and resins.“ Glands secreting poisonous compounds (tox-
albumins ; carbylamines (?)) occur in the mouth of snakes, in the
skin of the toad, on the feet of scolopendras. Crustaceans and
beetles produce chitin, spiders and caterpillars fibroin. Carabide
secrete butyric, bees and ants formic acid. Especial interest is
connected with the anal glands of the skunk,‘? musk deer,
beaver, and civet. Again, the livers of different animals vary
in their actions, as must be inferred from the composition
of bile and urine. Dogs can produce ethyl sulphide (Abel)
and an oxyquinoline-carboxylic acid (Kvretschy), birds ornithuric
acid;°) the bile of geese contains chenotaurocholic, that of
swine hyotaurocholic acid. Birds and snakes produce more
uric acid than urea, while the reverse is the case in mam-
malia. The products of metabolism exhibit, thus, considerable
discrepancies.)

This very brief survey may bear sufficient testimony to the
variety of the chemical work of organisms, and to the special
adaptation of the plasmic energy to the most varied chemical
performances. Few authors have thus far tried to propound an

(1) An elaborate treatise on the resin-producing glands of the conifers was
recently published by H. Mayr, formerly professor in the Imperial University of
Japan.

(2) The yellow liquid secreted by the skunk-glands and ejected in self-defence is
a primitive and effective weapon, having a most repulsive odour. It contains 16 per
cent of sulphur (Szarts), and is probably a mixture of several mercaptans with one
or more nitrogenous compounds.

(3) This acid, a dibenzoyl-ornithin, is secreted after introduction of benzoic acid ;
ornithin again is probably a’diamido-valerianic acid (Faff¢).

(4) Also, pathological processes may be caused by administration of certain
compounds only in certain animals; thus phloridzin will produce diabetes in dogs,
but not in rabbits or frogs.

THE ENERGY OF THE LIVING PROTOPLASM. 165

acceptable hypothesis for this activity. Nagel in his attempt to
explain the fermentative activity of the yeast-cell assumes a
certain kind of motion in the living protoplasm, imparted to the
glucose molecules, loosening affinities, and leading to dis-
ruption into carbon dioxide and alcohol. Mc. Laughlin went
still farther, applying the laws of oscillations established by
physicists to the action of bacteria in infectious diseases.’ But
neither Nageli nor Mc. Laughlin touched the question why such
energetic motions stop at once on the death of a cell, although
the conclusion that the living protoplasm must consist of
easily changeable, labile proteids was close enough at hand.
Mc. Laughlin expresses his views upon the fermentative action of
bacteria and yeast-cells in the following words: ‘‘ The distinctive
energy or waves of a cell can influence those substances only,
whose waves bear a certain relationship to those of a yeast-cell ;
and they must be equal in their periods, direction, and, perhaps,
in other characteristics, before those on one side can influence
those on the other. The nature of this influence will again
depend on whether the two sets of waves coincide in trough and
crest. If they do, the waves will supplement each other and
their amplitude will be enlarged; if they do not, they will
antagonise each other, and their amplitude will be diminished, or,
it may be, the waves will be destroyed by mutual antagonism ; it
will be remembered that all this occurs in waves of sound, of
light, and of water, and, if analogy has any merit, it can occur in
waves of molecular energy.”” Such considerations appear to be
justified, but there exist doubtless other influences which may
modify the expected result, such as the configuration of the
plasmic proteids and the tectonic of the plasmic apparatus (see
pag. 163, foot note). Further, it must be borne in mind that the
wave motions starting from the living protoplasm are of a far
more energetic nature than the molecular oscillations of any
other material in the cell.

The chemical performances of living organisms could not fail

(1) Fermentation, Infection, and Immunity, Austin, 1892. This treatise starts,
however, on several occasions from assertions which recently have been proved to be
incorrect. ‘The toxalbumins of the infectious diseases are not produced from animal
proteids by fermentative actions, but are secretions of the bacterial protoplasm which
are formed even in culture media devoid of proteids.

166 THE ENERGY OF THE LIVING PROTOPLASM.

to arouse the envy of the chemist and instigate him to try to
accomplish the same ends. To these endeavours we owe a long
series of interesting syntheses since the artificial preparation of
urea by Wohler in the year 1828. Carbon and hydrogen were
united in the electric arc by Berthelot ; acetylene, thus formed,
leads, among other things, to ethylene, ethyl alcohol, acetic acid,
aldehyde, acetone, and glyoxal. Acetylene yields at low red
heat benzene, from which, again, numerous derivatives can be
obtained. Acetone, further, may easily be converted into
trimethyl-benzene, aldehyde into trimethyl-pyridine ; from pyri-
dine, again, conine may be reached (Ladenburg). Glyoxal. leads,
by way of its cyanhydrin, to tartaric acid.

Carbon can be united with aluminium and this product
yields, by decomposition with water, methane (Mozssan). Methane,
again, yields methyl alcohol and formaldehyde, while the latter
yields by condensation several kinds of sugars (O. Loew).
Formic acid can be obtained by the action of sodium upon moist
carbon dioxide (H. Kolbe), or by the action of iron filings upon
bisulphide of carbon in presence of water in sealed tubes at
to0o° (O. Loew). Oxalic acid results by passing dry carbon
dioxide over hot sodium amalgam (LE. Drechsel), or by treating
the product of reduction of bisulphide of carbon by sodium
amalgam with fusing potash or boiling baryta water (O. Loew).
From a mixture of oxalic ester and acetic ester aconitic acid can
be obtained (L. Claisen and E. Hort); from acetone and oxalic
ester, oxytoluic acid, and from this an anthracene derivative,
(dimethyl-anthrarufin) was obtained (Claisen). From malonic
ester phloroglucin can be reached (A. Baeyer), from succinic
ester hydroquinone, from malic acid oxynicotinic acid and
daphnetin (Pechmann).

These and similar synthetic processes, are, however, for the
most part only possible by the application of powerful agents,
such as strong bases or acids, sodium alcoholate, sodium
amalgam, chloride of zinc, etc., and partly by the aid of high
temperature; while no light is thrown upon the special method
followed by the living protoplasm of plant-cells, a material consist-
ing of proteids of neutral reaction or nearly so, and very easily
changed by any substance with powerful affinities. The chemical
agency consists here merely of specific waves. We are acquaint-

THE ENERGY OF THE LIVING PROTOPLASM. 167

ed with chemical actions caused by waves of heat, light, elect-
ricity, and even by the waves of sound in some instances (explo-
sion of iodide of nitrogen), but concerning the related phenomenon
of chemical action set up by oscillating motion of the labile
(unstable) position of atoms, our knowledge is but very scanty.
Such processes are of the kind termed katalytic actions. This
expression was first used by Berzelius to designate chemical
phenomena apparently caused by mere contact with a certain
substance. The unsatisfactory definition of Berzelius, and the
denomination of certain processes as katalytic which in reality
were not such at all, implied a misunderstanding, and when Liebig
ridiculed the idea of Berzelius, nobody seemed to dare any more
to speak of this interesting group of phenomena and to explain
satisfactorily, for example, the wonderful activity of platinum
black.)

Katalytic actions exist, however, but they are not the result
of a mere contact, as Berzelius believed, but of a certain amount
of energy being conveyed, whereby the katalyser remains
entirely intact. If, however, the apparently katalytically acting
substance undergoes intermediary chemical changes with final
regeneration, the process is not a katalytic one. Such fseudo-
katalytic actions are, for example, the role of nitric oxide in the
manufacture of sulphuric acid, that of cobaltic oxide in the
development of oxygen from chloride of lime, the action of

(1) The action of light may consist in bringing about (a) combinations, as that
between chlorine and hydrogen, or (b) disruptions ; bisulphide of carbon is split into
sulphur and a lower sulphide (O. Loew); aqueous sulphurous acid is split into sulphur
and sulphuric acid (O. Loew); nitric acid into oxygen and nitrous acid; silver salts
are reduced to metal; (c) reductions of organic compounds: in alcoholic solution
quinone is changed into hydroquinone, nitrobenzene into aniline (Ciamician and
Silber), benzil into benzil-benzoin, phenanthrene-quinone into phenanthrene-hydro-
quinone, a portion of the alcohol present being converted into aldehyde; (d) poly-
merisation and condensation; thymoquinone, anethol, phenyl-naphtoquinone are
polymerised ; monobromacetylene is converted into tribrombenzene, propargylic acid
into trimesic acid (Ber. D. Chem. Ges. 27, 958).

(2) The great resemblance of the chemical activity of living cells to katalytic
actions was recognised more than thirty years ago by the great physiologist, C. Ludwig.
Also C. Lehmann in his “ Lehrb. d. physiolog, Chem.,”’ and, later on, Tvaube and
Stohmann expressed themselves in the same sense. But explanation of this katalytic
action was wanting.

(3) Even now-a-days, isolated voices are raised in the sense of Liebig. See
H. Macleod’s lecture at the meeting of the British Assoc. in Edinburgh 1892.

168 THE ENERGY OF THE LIVING PROTOPLASM.

aluminium chloride in the synthesis of hydrocarbons, that of
zinc chloride in the transformation of glycol into aldehyde, and
the accelerating action of iron and copper salts upon the
oxidation of phenol by peroxide of hydrogen.“

The genuine katalytic actions may be divided into three
groups:

1. Katalysis by labile organic compounds,

2. Katalysis by mineral acids, caustic lyes, and certain salts,

3. Katalysis by finely divided metals.

In regard to the first group may be mentioned the conversion
of dicyanogen into oxamide by a dilute solution of ethylic
aldehyde, a reaction observed by Liebig ; the transformation of
thiourea into the isomeric ammonium thiocyanate by an alcoholic
solution of ethyl nitrite (Claus) ; the facilitating action of acetic
ester upon the combination of hydrocyanic with hydrochloric acid
(Claisen and Mathews). Maleic acid transforms ketazines into the
isomeric pyrazolines with liberation of heat, whilst fumaric
acid can only accomplish this at a temperature of 100.° To this
group of actions belongs evidently not only the action of the
living cells, but also the action of the enzymes, which have a
high degree of lability and betray, by their easily passing into an
inactive state, as well as by their proteic nature, a certain rela-
tion to the proteids of the living protoplasm. The fact that
dilute formic aldehyde destroys their activity even in perfectly
neutral solution at the ordinary temperature, makes the presence
of highly labile amido-groups probable.)

(1) Chem. Centralbl. 1885, p. 224. In certain cases the apparently katalytically
acting substance is mainly a suitable medium to bring two compounds into more
intimate contact with each other, or by combining with one of two compounds pre-
sent to help to loosen somewhat certain affinities in the molecule, and thereby bring
about combination with the second compound.

(2) Curtius and Foersterling, Ber. D. Chem, Ges. 27,770. F. Stohmann deter-
mined the thermic value of maleic acid to be 326.3 cal. and that of fumaric to be 319.7 ;
the latter possesses, therefore, less energy than the former. Stohmann recognised by
direct experiment, in determining with great exactness the heat ofcombustion, that
labile compounds always have a higher thermic value than the isomeric stable ones.

(3) O. Loew, Jahresber. f. Thierchem. 1888. I have observed also that a solution
of prussic acid of 25 °/, destroys in twelve hours the diastatic but not the pro-
teolytic enzyme of the pancreas (Pfliig. Arch. 27, 208). Cf. also Schdry and Fichter,
Jahresber f. Thierchem. 5, 269. Certain bacterial enzymes are rendered inactive by
sulphuretted hydrogen (Fermi and Bernossi. 1894). Chloroform retards enzyme-action
(Salkowski). According to Arsonvale (1894) enzymes are rendered inactive by a
temperature of—150°C. .

THE ENERGY OF THE LIVING PROTOPLASM. 169

In the second group of katalytic actions may be counted the
transformation of maleic into the isomeric fumaric acid by mineral
acids, of maleic ester into fumaric ester by contact with hydro-
chloric acid at the ordinary temperature (Skvaup) and of citra-
conic into mesaconic acid (Delisle, Franz) ; of hydromellitic into
isohydromellitic acid by hydrochloric acid (Baeyer) ; of dihydro-
terephthalic into an isomeric acid bya solution of caustic soda,
the formation of paramidophenol from phenyl-hydroxylamine
(E. Bamberger) and that of paranitroso-compounds from nitro-
samines by acids. Also the transformation of oleic into elaidic
acid by nitrous acid deserves mentioning.

In connection with the third group we mention the following
observations: platinum black brings about an oxidation of
hydrogen, of alcohols, and of various other compounds ; it unites
sulphur dioxide with dry oxygen to form sulphur trioxide; it
combines hydrogen with hydrocyanic acid into methylamine at
110° (Debus) ; it transforms a mixture of nitric oxide and hydrogen
into ammonia and water; it accelerates the decomposition of
hydroxylamine in presence of caustic potash; it decomposes
peroxide of hydrogen energetically; it transforms ozone into
common oxygen (Mulder) ; it decomposes azoimide into ammonia
and nitrous oxide (O. Loew), and nitrososulphates into sulphates
and nitrous oxide (Pelouze).

Finely divided iridium or rhodium decomposes formic acid
into carbon dioxide and hydrogen (Deville and Debray) ; palla-
dium powder effects an oxidation of hypophosphorous acid with
liberation of hydrogen ;“ finely divided copper incites a rapid
decomposition of formic aldehyde by caustic potash with libera-
tion of hydrogen,'” it also decomposes diazobenzene chloride into
nitrogen and chlorbenzene at low temperatures.) Zinc filings
condense at 100° acetaldehyde to aldol and crotonaldehyde.

All these actions of finely divided metals can be best ex-
plained by the assumption that a modification of heat waves
takes place in such a manner that this energy can now pass
more easily into chemical energy. With platinum black this

(1) Engel, Compt. rend. 116, 786. The chemical explanation given by this
author is certainly incorrect.

(2) O. Loew, Ber. D. Chem. Ges. 20, 145.

(3) Gattermann, ibid. 2%, 1218 and 25, togr, footnote,

170 THE ENERGY OF THE LIVING PROTOPLASM.

modification is carried still farther by the dense layer of oxygen
surrounding the metallic particles.'”)

The katalytic action of mineral acids is evidently also
due to a certain motion in the molecules.'?) A motion of different
character, however, must be assumed in organic compounds of
a certain lability (cf. Chap. III). Usually the katalytic reac-
tions are exothermic ones, but in special cases, where other
agencies render their aid, also endothermic, as in the action of
the chlorophyll-bodies upon carbonic acid in sunlight.

The katalytic powers of platinum-black are evidently of a
very inferior character, compared with the faculties of living
protoplasm,—we notice above all an entire want of condensing
influence—but in regard to simpler reactions a parallelism can
nevertheless be shown to exist, of which I here adduce some
further proof. One of the most general synthetical processes is
the formation of fat from glucose in living cells, a process
consisting in condensation and reduction:

3 Ce it. 710,—16'O— Cy geble OF
(stearic acid)
Cs Hi2 O6-+4H=2 C, Hs O,
glycerol)

The remarkable transformation of sugar into the higher
fatty acids has thus far not yet been imitated, but we can at least
obtain lower fatty acids of rancid odour if we mix a 10 % glucose
solution with about half its weight of active platinum-black ; the
odour will be perceptible after 1-3 hours and increase gradually.
The main action, of course, is a direct oxidation, but simul-
taneously a reduction is going on in other molecules which yield
up a part of their oxygen also to glucose molecules under the
influence of platinum-black.) Neither levulose nor cane sugar
will yield this result. Platinum-black, also, deprived of its
absorbed oxygen in one way or other, is inactive in this regard.

(x) Platinum-black can absorb 800 times its own volume of oxygen (Doebereiner),
but this alone would not suffice to explain its oxidising power, compressed oxygen
possessing no increased energy at ordinary temperatures. Cf. also O. Loew, Ber. D.
Chem. Ges. 23, 290 and 677, where I gave first the above explanation of the katalytic
action of platinum black.

(2) We will not enter here upon the most recent views in regard to the action of
acids, more light being stil! necessary.

(3) O. Loew, Ber. Deutsch. Chem. Ges. 23, 865.

THE ENERGY OF THE LIVING PROTOPLASM. 1G

If we add, for example, to the mixture mentioned, sodium
carbonate, or boil the mixture with addition of calcium car-
bonate, no trace of the rancid smell will be noticed after addition
of a mineral acid; the absorbed oxygen is here too quickly used
up for oxidation.

Another process, hitherto unexplained, is the easy reduction
of nitrates to ammonia in the formation of proteids (cf. Chap. V).
As there exists no nascent hydrogen in the living cells, I had
long entertained the view that glucose under the katalytic
influence of the living protoplasm would be the reducing agent,
and recently have succeeded in imitating this reduction by
means of platinum-black. I heated in my first experiment a
solution of 3 g. potassium nitrate and 10 g. glucose, in 300 g.
water with rro g. platinum-black for 6 hours to 60-70° and found
that 45.6 % of the nitrogen of the nitrate had been converted
into ammonia. This reduction can even succeed at the
ordinary temperature. If 50 cc. of a mixture of a 5-10 % glucose
solution with 1-2 % calcium nitrate and 10g. platinum-black is
left to stand for 4-5 days in a closed flask, we observe upon
supersaturation with caustic lye a strong ammoniacal odour, while
in a control experiment without the platinum no trace is obser-
vable.”) If we modify the experiment by increasing the amount
of the nitrate to three times that of the sugar, we find on heating
that the acid reaction generated at first soon diminishes and
finally gives way to an alkaline one, and then the ammonia pre-
viously formed becomes very perceptible by its odour.“) The
platinum-black imparts to the hydrogen atoms of the glucose
increased motions, thereby loosening the existing affinities, and
awakening others, whereby a reduction of the nitrate is caused,
1.¢., an exchange of oxygen and hydrogen between the nitrate and
the sugar.

In a similar way, chlorates are reduced to chlorides;
sulphates however resist its action and require evidently more

(1) Other control experiments were also made, which proved beyond any doubt
that only the platinum-black itself and not any oxidation product or bacterial action
was the cause of the formation of ammonia.

(2) Ifthe mixture is rendered alkaline before the platinum-black is added, no
ammonia will be produced, a result which finds its explanation in what has been
mentioned above.

172 THE ENERGY OF THE LIVING PROTOPLASM.

energy ; but at least with a certain sulpho-acid, the combination
of formic aldehyde with acid sodium sulphite, I succeeded in
effecting reduction; for this in presence of sodium carbonate
yielded by moderately heating with platinum-black, small
quantities of sodium sulphide,’) while the odour of methyl
mercaptan became perceptible.

The katalytic action of platinum-black is manifested still
in another case of physiological interest. If moistened with
pure caustic lye and exposed to pure air it forms from nitrogen
and water nitrous acid and ammonia to a small extent.” Such
a process may take place when nitrogen is assimilated by legumi-
nous plants whose roots have entered into symbiosis with
certain bacteria. The reverse process can also be katalytically
accomplished ;~ for a neutral solution of ammonium nitrite,
which is only decomposed by application of /eat, will in presence
of platinum-black show a continuous development of nitrogen at
the ordinary temperature. A mixture of 6 g. ammonium sulphate
with its equivalent quantity of potassium nitrite dissolved in
130 cc. water developed after addition of 20 g. platinum-black in
24 hours Igt cc. nitrogen, and in 5 daysas much as 768 cc., at
15° and 723 mm. barometric pressure. A similar process takes
place in cases of putrefaction when nitrates are present, where
the protoplasm of the bacteria present acts upon the nitrite
of ammonia formed and liberates nitrogen.

In all the cases described here the platinum-black remains
exactly what it was; it does not act by virtue of any chemical
affinities, but only by a specific mode of motion, Analogous to
this is the action of living protoplasm; it remains what it is
while it produces various chemical changes in compounds that
come into contact with it. If it acted by means of chemical
affinities, it would undergo a change, and that would mean the
death of a cell when that change amounted to more than slight
traces inthe unit of time. There can be no doubt that the
active principle is an intense and ceaseless motion of atoms intimately

(1) We might express this result by the following equation :
2 CH20H.SO3Na+4NazC03=Na2S+S03Naz+ HCOONa+5NaHCO;
Compare O. Loew, Ber. Deutsch. Chem. Ges. 23, 3125.
(2) O. Loew, ibid. 23, 1443.
(3) O. Loew, ibid. 3019.

THE ENERGY OF THE LIVING PROTOPLASM. L732

connected with the chemical constitution of the protetds composing
the living protoplasm (cf. Chap. III and V), and intensified by the

respiration process (cf. the following chapter).
Grant Allen in his admirable treatise ‘‘ Force and Energy”

gives the following

“Table of kinetic energies :”

Separative Separative Separative Separative
: , molar motion. molecular MEOTICERIOTOnE electrical
Separative. |(Ina body raised motion. Z motion,
from the earth’s| (In a body torn | (In chemical | ({n an electrical
surface). apart). decomposition). machine).
Aggregative Aggregative Aggregative Aggregative
: molar motion. molecular atomic motion. electrical
Aggregative. ; motion. P :
(In a falling (In a body cool- (In chemical motion,
body). ing). combination), (In lightning).
Continuous Continuous Continuous Continuous
ue molar motion. molecular . : electrical
Continuous. i motion. atomic motion ;
(In a top or in (Motion in motion.
a planet). heat). Unknown.() (In magnet).

Now, the energy produced by continuous atomic motion, for
which Allen could not cite an example, must be identical with

that displayed by atoms in labile position.

The foremost ex-

ample of such energy is represented by Plasmic Energy.

(Cis ieainin WAIL.

Respiration.

Philosophers who have recognised respiration as the princi-
pal foundation for all the other vital functions have frequently
compared the living organism with a machine using coal; in
both cases a liberation of energy by combustion and the applica-
tion of more or less of it for various useful performances are
taking place. This comparison, however, is only admissible

(1) The italics are mine.

174 THE ENERGY OF THE LIVING PROTOPLASM.

within a very narrow compass, since the difference becomes great
with an increased supply of oxygen. Hereby, the coal will burn
with much greater vigour, whilst respiration will not be in-
tensified, because the amount of oxygen needed is regulated by
the living cell (Pfliger).

The respiratory intensity not only exhibits great differences
in the various species of organisms,’ but also in different organs
of one and the same organism.

The intensity is greater in flowers than in leaves and
roots,’?) greater in leaves than in stem and fruits (Saussure),
greater in light-plants than in shade-plants (Adolf Mayer),
greater in shoots of oil-seeds than in those of starch-seeds
(Godlewski), greater in air-plants than in hydrophytes (Boehm),
in the cat double what it is in the sheep (Mumck).

While increase of pressure of oxygen will not influence the
intensity of respiration, the effect of rising temperature is, on
the other hand, very considerable, the activity of the protoplasm
being thereby greatly enhanced. At o° the respiration of plants
is very slight, at 17-20° it is already twenty times more intense,
and still gradually increases with the temperature nearly up to
that of death. Cold-blooded animals respire less than warm-
blooded," while animals in hybernation exhibit a lower tempe-
rature and respire less than those in activity.

Heat, visible motion, chemical action, and, in certain cases,

(t) To-day it seems hardly credible that Liebig still maintained that chloro-
phyll-bearing plants will not carry on respiration, although Saussure had positively
proved to the contrary as early as 1805! Another erroneous conception, viz., that
the blood is the principal seat of respiratory oxidation in animals was corrected in
the year 1868 by Pfliiger, who has recently told us what enormous labour for years it
took him to convince physiologists of the truth that oxidations in animals are
accomplished by the cells of the various organs.

(2) The root-ends of young plants of Vicia Faba consume in 24 hours 5 per
cent of their dry matter (Palladin). Young rootlets and especially root-hairs have
an energetic respiration, while the interior of large roots respires certainly much less
than leaves. Not only the restricted access of air but also the lower temperature to
which usually roots are exposed in greater depth will under ordinary conditions
lower the intensity of respiration.

(3) The intensity in frogs and earthworms is only about one tenth that in the
dog; that of higher plants is generally found less than that of warm-blooded animals,
in many cases also less than that of cold-blooded, if equal weights of dry matter are
considered. It is with pea-shoots about one half that of the frog, while in well
nourrished mould fungi it considerably surpasses even that of mammalia.

THE ENERGY OF THE LIVING PROTOPLASM. 7

electrical phenomena and light are the forms of energy resulting.
A stratum of germinating barley, 15 cm. high, at an air tempera-
ture of 7°, reaches, after a few days, a temperature of 18°. One
grm. of a stalk of Brassica liberates by respiration in one hour an
amount of energy equal to 132.1 gram-meter, amounting for one
cell per minute to 2.2 milligram-millimeter (Rodewald). The
greater part of the liberated energy assumes the form of heat
and is dissipated. Bees produce so much heat that their hives
show even in winter a temperature of 30°. Rabbits, with a blood
temperature of 38°, consume per kilo. and per hour 0.g14 grm.
oxygen, while chickens, with one of 43.9°, consume 1.186 grm. of
it. Man produces every hour as much heat as would raise the
temperature of an equal amount of water 1.4 degrees.

Every increase of muscular exertion requires an increase in
respiration ; the work done by a muscle corresponds to about 33
per cent of the energy yielded by respiration, while in a steam-
engine only ro per cent of the caloric value is secured in the
form of work.’ An animal, therefore, considered as a machine,
works economically (Zuntz).

Even the finest currents in the protoplasm depend upon the
presence of oxygen, as Kiihne has shown, and the increased work
connected with a rapid development of shoots and embryos, or
with the transportation of starch in plants requires also an
increase of respiration. Further, the dependence of the produc-
tion of light by fungi and by various animals upon respiration
has been proved, and there can be no doubt also that it is the
same with regard to the production of electricity in the nervous
system and in the electrical organs of fishes.

The function of respiration consists, not in all cases, however,
in merely yielding kinetic energy ; it serves also to prepare useful

(1) The knowledge of the caloric values of the different compounds encountered
in organisms being of high physiological interest, Stohmann has determined them
with great exactness recently. We mention a few data. The caloric values of 1
gram amounts to, in:

Peal UU Ure lintels etelelsleleislelele ciasiticisieleeieieieen (55735. 20 Cale
PE DLOUCMelselststeiieeeietsieinl esieisls) siele'slevelelsieisiele) 532000" 35
BCUCIetelereteieiiatlaeistelelslcieleteceidieitetinieisn sistsian | O15 33:0) 45
At) ee etes einisiciele sree eves Sielpieisieleivieieiviccs ccfee 'Q;50G:0) 55
GUNICOS CaereloleleralelnloletslelsieVeloislolelsielsieicieiolele eieieis) 3374250) 8

176 THE ENERGY OF THE LIVING PROTOPLASM.

oxidation products, as carbohydrates from fats in plants,"") and it
helps to prepare from different materials the necessary starting
groups for protein formation (cf. Chap. V).

While a unicellular organism obtains the necessary oxygen
by a simple diffusion process, more or less complicated contri-
vances are necessary to provide the interior cells of multicellular
organisms with air. Stomata, lenticells, and intercellular spaces
serve this purpose in the vegetable kingdom; tracheids, gills,
lungs, hemoglobin, in the animal kingdom. Movements of the
abdomen and thorax in insects, of the gills in aquatic animals,
of the chest in lung-bearing animals, maintain the exchange of
oxygen against the resulting carbon dioxide. Certain insects, as
the larve of the Lzbellulidg, and even a fish (Cobitis fossilis), exhibit
the remarkable exception, of carrying on their respiration by the
intestines, which are provided for this purpose with innumerable
blood vessels.

Not all organisms, however, produce their physiological
energy by respiration ; the fermentative organisms gain it with-
out the aid of oxygen, by decomposition of organic matter,
Bacteria can utilise various hydroxy-acids, proteids, poly-
valent alcohols and sugars, while the yeasts certain kinds of
sugar only. We have here the case of the so-called tntramolecular
vesptration before us, one which yields considerably less energy
than normal respiration. Also certain animals can for a limited

(1) The formation of organic acids is also due to respiration, viz., to an imper-
fect oxidation of sugar in most cases. An interesting example is furnished by the
flowers of Ipomea triloba, which are blue in the morning and remain so during
cold foggy, and rainy days, but turn ved on warm bright days. Since this change
from blue to red can also be easily accomplished by acids, we must assume that the
increased respiration on warm days causes the production of acids.

(2) The fact that bacteria can be deprived of their fermentative faculties,
without their life being impaired and thus be turned from anaérobs into obligate
aérobs, has led me to the view, that there exists a special organoid in those organisms
endowed with fermentative action. This would, to a certain degree, be analogous
to the chloroplasts of green plants; the latter prepares suitable material for growth
from carbonic acid, while the former prepares it by decomposing various organic
matters. A small fraction of these fermenting compounds does not appear in form of
fermentation products, but in that of new cells of the fermenting organism. We see
therefore, here also a double part played, viz, that of liberating energy and that of
preparing the necessary groups for protein formation. Cf. O. Loew, Centralbl f.
Bacteriologie 9, Nr. 22.

THE ENERGY OF THE LIVING PROTOPLASM. 177

time subsist on intramolecular respiration, as Pfliiger has observ-
ed in frogs, and Bunge in worms."?

Although the comparison of respiration with direct com-
bustion was close at hand, still there were many mysteries
involved and many questions unsolved, in regard to the laws and
to the causation of that energetic combustion going on at rela-
tively low temperatures in a substance containing 75 per cent
water and more, and yet remaining apparently intact, while
effecting the union of the resorbed food with free oxygen. That
the protoplasm remains alive while this fierce and destructive
oxidation is carried on, appears the more notable as it is known
how easily all kinds of living cells are killed by oxidising media
in high dilutions, as by peroxide of hydrogen or by potassium
permanganate.

The views propounded by various authors exhibit conside-
rable discrepancy. The oldest hypothesis, that of Schdnbein,
assuming the formation of ozone, had soon to be discarded. But
nevertheless, the idea that the common oxygen had to be changed
into an active form or modification before it could unite with the
compounds in the cell, prevails also in the other theories. ‘The
supposed activifying process consisted in the splitting of the
oxygen molecule into its two atoms with free affinities.) Hoppe-
Seyler supposed that the living cells produce hydrogen, which in
its nascent state could accomplish the ‘‘ activifying ”’ process by
combining with one of the atoms and setting the other free. But
it was objected that hydrogen ought to make its appearance at the
moment oxygen is being withdrawn. This was, however, never
observed. Germinating seeds can exist one day, certain worms
(Ascaris) even 5-7 days, alive in absence of oxygen, but no trace
of hydrogen is evolved by these organisms during that time.)
Reinke holds that there exist in the living cells easily oxidisable

?

Organic matters, ‘‘ autoxidisers,’ which are capable of suffering

(1) With insufficiency of oxygen, animals will show albumen, glucose, and lactic
acip in the urine (Araki), also an increase of oxalic acid (Reale and Boeri).

(2) This process would require a large amount of energy. Heat alone can
accomplish it only at a temperature not lower than 1400° C. (Troost and Haute-
feuille).

(3) M. Traube contends, moreover, that nascent hydrogen can activify oxygen ;
it can merely produce peroxide of hydrogen.

178 THE ENERGY OF THE LIVING PROTOPLASM.

oxidation in contact with molecular oxygen, so as to produce
peroxide of hydrogen, which under the influence of enzymes would
bring about powerful oxidations.”) But Reinke omitted to prove
that the supposed ‘‘ autoxidisers’’ would be really capable of
inducing oxidations of sugar or fat, while as regards that perox-
ide its absence in plant-cells has been proved by Th. Bokorny'”)
and by W. Pfeffer.©) Neither could it be discovered in animal
cells. Retnke’s view moreover could not explain how respiration,
throughout such a wide range, is independent of the amount of
oxygen present. But the belief that there exist in many plants
easily oxidisable compounds is no doubt true. Many plant juices
acquire soon a reddish or brown coloration, if exposed to air, a
phenomenon which illustrates how different a course is that
taken by respiration proper, since these same compounds were
no doubt formed in the protoplasm and afterwards gradually
secreted into the vacuole. Numerous plants, however, do not
yield a juice of like behaviour, nor has this been observed in
animal juices. Iczinke’s observations have therefore no connec-
tion with the respiration process.

The theory of Traube) assumes the existence of oxidising
enzymes, which would act as transporters of oxygen, somewhat
like nitric oxide in the manufacture of sulphuric acid.©) The
occurrence of such enzymes was not proved by Tvaube, but has
been demonstrated a year ago beyond any doubt by Toyonaga, who,
at my request, made a series of investigations. These enzymes
are the cause of the darkening of the juices of many plants when
easily oxidisable matters are present, as in potatoes, in the roots

(1) Botan. Zeitg. 1883. Nr. 5 and 6.

(2) Ber. Deutsch. Chem. Ges. 24, r100 and 1848. Pringsh. Jahrb. 17, 347.

(3) Ber. Sachs. Akad. d. Wiss, 1889. Recently the presence of Hz Oz in various
plants has again been asserted by Bach, but his reaction was unreliable (cf. Cho, this
bulletin, and his conclusion not justified).

(4) Theorie der Fermentwirkungen, Berlin, 1858. Virch. Arch. 24, 386. Ber.
D. Chem. Ges. 10, 984.

(5) Analogous processes are the action of ferrous salts in the oxidation of
tartaric acid in sunlight (Vries), the oxidation of aniline by a slight trace of
ammonium vanadate (Bull. soc. chim. 45, 309) or the oxidation of nitrogenous com-
pounds when added to a solution of oxide of copper in ammonia exposed to air (O.
Loew, Z. Biol. (1878); Journ. f. prakt. Chem. 1878). Another example, sometimes
cited, viz., the oxidation of indigo-white and its regeneration by alkaline glucose
solution is for obvious reasons not well suited for comparison in this connection.

THE ENERGY OF THE LIVING PROTOPLASM. 179

of Daucus, Beta, Lactuca, and Taraxacum. Among the alge
Zygnema may be mentioned, whose fresh juice turns black, and
among the fungi Boletus luridus, whose freshly cut surface turns
blue in contact with air.) If, however, oxidising enzymes
are present while those oxidisable compounds are wanting, then
the existence of the former can be shown by adding guaiacum
solution or hydroquinone, pyrocatechin, or pyrogallol. Guaiacum
will yield a blue color, the latter a brown or black one.
Schénbein observed this blue reaction in various seeds, and
especially well in those of Cunara ;°?) Molisch in the secretions of
various roots, as those of Pisum, Brassica, Cucurbita, Lepidium,
Scorzonera and Neottia.2) Some observations of Pfeffer have
made it probable that the supposed enzymes are contained in
the protoplasm itself, and come therefore into intimate contact
with the easily oxidisable compounds contained in the cell-sap only
after the death of the cells, when the osmotic properties of the
tonoplast have gone.“) ‘That the blue colouration of guaiacum is
also due to the action of an enzyme is supported by the fact that
fresh diastase can produce the same reaction.©) I have further
observed that an enzyme-like compound of proteid nature can be
precipitated from potato juice by alcohol, and that it loses its
quality of blackening hydroquinone upon drying in the exsiccator.
I have also observed that the blackening of Zygnema never
takes place after killing the cells with sulphuric acid or absolute
alcohol. Toyonaga determined the temperature at which the
potato lost its peculiar action upon hydroquinone, pyrogallol, and
pyrocatechin, and found it to be 73°. Mercuric chloride destroyed
its qualities at 55° in one hour, formic aldehyde (5 %) after 2

(t) Also the cambial sap of the conifers turns gradually brown in contact with
air. The turning brown of blossoms and green leaves which sets in after death,
belongs to the same group of phenomena. The existence in plants of oxidising
enzymes was recently also shown by Bertrand C.r. 120.

(2) Journ. prakt Chem., 88 and 105; Gmelin-Kraut, 1. 2. p. 456.

(3) Wien, Akad. Ber. 1887.

(4) The compound yielding the colouration of potato juice can be extracted
with alcohol; it turns dark brown in contact with Fe Cl3.

(5) &. Schéne (Z. analyt. Chem., 33, 159) observed that old guaiacum tincture
yields a greenish blue colouration with diastase alone; fresh tincture, however,
wants the addition of hydrogen peroxide. The substance yielding the blue product
is guaiaconic acid, C20 Hz4 Os (Liicker).

180 THE ENERGY OF THE LIVING PROTOPLASM.

days standing, almost completely; dilute sulphuric acid and
caustic potassa acted in the same way, so that there can be
hardly any doubt that the active principle is an enzyme.

Oxidising enzymes appear to be present also in the animal
body. Saliva produces a blue colour with Wurster’s reagent
(tetramethylparaphenylene-diamine), a fact erroneously taken to
be a proof of the presence of hydrogen peroxide; it produces
further a brown colouration with hydroquinone. E. Salkowski
found in the blood, Faguet and also Yamagiwa in the lungs,
kidneys, and muscles, enzyme-like agencies, capable of producing
small quantities of benzoic acid from benzyl alcohol and of
salicylic acid from its aldehyde.) But such actions of enzymes
are always of a very narrow compass; they belong to very weak
oxidations of certain benzene compounds, but never to an attack
upon, to say nothing of a complete combustion of, sugar or fat.
We must therefore reject also the theory of Tvaube as wholly
unsatisfactory.

It appears singular that the authors mentioned all ignored
just the most important condition for respiration, 7.c., the /iving
state of the protoplasm. ‘Taking a plain chemical start and leaving
physiology aside, they endeavored to reach a satisfactory expla-
nation, biased by the idea that the albuminous compounds the
chemist studies in his vials are the same as those composing
living matter. This erroneous conception still governs the
minds of many, and by them respiration will never be com-
prehended.

It was Pfliigey who in the year 1876 drew the inference in
plain and forcible logic that the proteids of the diving protoplasm
are different from those of the dead, and that their chemical
change into the indifferent common proteids signifies the death
of the cells. Only those chemical qualities exhibited by the
living cells can induce respiratory activity. ‘‘ Oxygen is not
made active, but the proteids of the living cells have the
activity.”’ Thus, a new foundation was gained, but few physio-
logists took notice of it; above all, however, Nenck: assented.
Detmer modified Pfliiger’s view, assuming a continuous dissociation

(1) Jahresb. f. Thierchem., 22, 387. Also Réhmann and Spitzer: Ber. Chem.
Ges., 28, 567.

THE ENERGY OF THE LIVING PROTOPLASM. 181

of the “units of life,’ whereby respiration would be induced.”
The dissociation he assumed to be followed by regeneration.
This barbarous idea of dissociation, however, would be incom-
patible with the great sensitiveness of the living protoplasm ; dis-
sociation would mean simply death and nothing else.

But how can the living protoplasm, in spite of its extraordi-
nary sensibility, carry on such a powerful combustion process,
surpassing in energy the action of nitric acid, without being
injured? Can we hold it possible that the oxygen is first
activified before it oxidises? It would seem to us that an
activified oxygen would have certainly other ways of action than
those observed within the living protoplasm. Respiration
exhibits like the well studied oxidations by various agents," its
own specific manner of oxidising. An animal capable of oxidis-
ing in 24 hours several hundred grams of starch (sugar) completely
to carbon dioxide and water,®) is unable to burn up a few grams
of oxalic or formic acid.“) It is even unable to oxidise I gram
of benzene completely to phenol. And while it destroys tyrosin,
quinaldin, and pyrrol, it attacks with difficulty hydroquinon,
phenylacetic acid, or naphthoic acid.

A series of investigations by Nencki, Mering, Baumann,
Salkowski, and others, led to the recognition that certain com-
pounds reappear in the urine unchanged, such as benzidine,
others in combination with sulphuric acid, as phenol, others
again (partially oxidised or not) as derivatives of glycocoll, like
picoline or benzoic acid, or of glucuronic acid, as tertiary alcohols
and thymol, or of cystin, as brombenzene; they may also re-
appear as uramido-compounds like sulphanilic acid, or taurin.

(1) Physiologie des Keimungsprocesses, Jena, 1880; Jahresb. Thierchem., 22.

(2) Potassium permanganate, nitric acid, hypochlorites, hydrogen peroxide, lead
peroxide, silver oxide, have all a specific oxidising action. The results may however
sometimes be modified by relatively small changes in the molecules to be oxidised,
as by the introduction of an acidic or alkylic radical.

(3) The question as to the intermediary products is of relatively small im-
portance. The formation of glucuronic acid in animals shows that to a very small extent
acids are produced, but formic and oxalic acid certainly only in slight degree. The
forerunner of carbon dioxide is probably the bivalent group HC-OH, but that cannot
be proved.

(4) Oxalate of sodium in non-lethal doses reappears in the urine with a loss of
only 7 per cent (Gaglio). Sodium formate reappears to the extent of 4-3 in the
urine (Gréhaut and Quinquaud),

182 THE ENERGY OF THE LIVING PROTOPLASM.

The lateral chains of aromatic ketones are oxidised to carboxy),
if the benzene ring does not contain a hydroxyl-group, otherwise
the entire compound will reappear in combination with sulphuric
or glucuronic acid in the urine, with the lateral chain preserved
(Nenckt). Some sulphur compounds will yield sulphuric acid,
while others will not.“

It is, further, of great interest that the oxidation of benzene
to phenol and to small quantities of pyrocatechin and hydro-
quinone in the animal body (Nencki and Gzacosa) is diminished by
the introduction of certain poisons.'”)

The difficulty in completely oxidising benzene derivatives of
a certain constitution is not only encountered in the animal but
also in the vegetal organism. Many plants accumulate tannins
and related compounds without ever utilising them again; in
most cases they are excreted and may by their repulsive taste
prevent depredations by various animals: only under certain
conditions may tannin be utilised again.©) Also certain alkaloids
in plants undergo no further metamorphosis) and have to be
looked upon as excreta. It is, further, very characteristic of
the oxidising faculties of plant-cells that certain compounds are
left unchanged, which, even by such a comparatively weak
oxidising agent as hydrogen peroxide, are attacked readily. Thus,
it was observed by Pfeffer that cyanin (a quinoline derivative),
introduced into plant-cells is not altered, while it is easily bleached
by the peroxide. Also certain compounds in the roots of Vicia
and of Trtanea Bogotensis are easily converted into brown oxida-

(1) Compounds of the general formula NHz—CO—S—R easily yield sulphuric
acid; thiophen or sulphonal do not (Smith).

(2) Nencki and Sieber (Pfliig. Arch. 31, 319). found, e.g., that while in the body
ofa healthy man 0.82 g. phenol was formed from 2 gram benzene, under normal
conditions, only 0.33 g. was formed if 2 gr. ethyl alcohol per kilo of body-weight were
administered at the same time. Also individual differences were noticed. Poisons
may also interfere with normal oxidising processes; thus, diamide in non-lethal doses
leads to the appearance of allantoin in the urine of dogs (Borisson, Z. physiol. Chem.,
19, 499)-

(3) Small kinds of Spirogyra will use up their tannin after a few weeks cultiva-
tion in a solution containing 0.5 °/o KHz PO4; 0.2 0/0 KH CO3 and traces of nitrate
and sulphate of calcium.

(4) Cf. the investigations of Leo Errera, Maistriau, and Clautriau, especially
those of the latter in the Bulletin Belge de Microscopie, 1894.

THE ENERGY OF THE LIVING PROTOPLASM. 183

tion products by it, while they remain colourless during their stay
in normal cells.“

How weak the oxidising power of the cells appears to be here
and then how energetic when sugar comes under action! We
search the entire domain of chemistry in vain for a single case of
the occurrence of complete combustion when an organic compound
in aqueous solution absorbs oxygen from the air; even the well
known energetic absorption of oxygen by alkaline solution of
pyrogallol does not lead to a complete combustion. Only the
action of free permanganic acid can exhibit similar results.

However, there exist cases where partial oxidations by
molecular oxygen can easily take place. We will mention the
transformation of aldehydes into acids, of hydrazo-benzene to
azobenzene, of indigo white into indigo blue. Also the be-
haviour of anthraquinone, oxindol, and amidophenols to common
oxygen may be cited. The total change here taking place
consists either in the entrance of one oxygen atom into the
molecule or inthe withdrawal of two hydrogen atoms. Other
compounds again acquire the property of absorbing free oxygen
through the presence of an alkali, as pyrogallol, pyrogalloquinone,
chrysarobin, furoin.%) Benzene acquires it by the presence of
aluminium chloride. In all these cases there exists evidently a

high degree of lability in certain hydrogen atoms leading to the
absorption of oxygen. This lability is due to their specific posi-
tion in the molecules.“ We observe under ordinary circum-
stances, however, no labile hydrogen atoms in the fatty acids
proper, but nevertheless the latter are easily burned up in the
cells. We must then logically conclude that contact with the
living protoplasm suffices to impart a state of lability to the atoms
in the molecules of the fatty acids, recalling the action of

(1) Ber. Sachs. Akad. d. Wiss., 1889, p. 493. These observations likewise proved
the absence of H2Oz2 in plant-cells.

(2) It is obvious that the energetic autoxidation of zinc ethyl, dimethyl arsine,
monobrom-acetylene and of the sodium compounds of ketones and aldehydes, which
burst into flame in contact with air cannot serve for comparison.

(3) A preliminary ‘ activifying’”’ of oxygen takes place here just as little as in
the living protoplasm,

(4) Lability of hydrogen linked to carbon may be of two kinds: one which
determines its easy exchange by certain metals, as in acetylene, the other which
causes its increased affinity for oxygen, as in aldehydes.

184 THE ENERGY OF THE LIVING PROTOPLASM.

platinum-black, which renders the alcohol molecule so labile that
it readily takes up oxygen. We must look upon both kinds of
phenomena as katalytic oxidations (cf. the foregoing chapter).'”

It is the CH.— group in the fatty acids and amido acids,
and the CH OH— group in ketoses, aldoses, and hydroxylated
acids that are most easily attacked. The carboxyl-group renders
the CH OH— group in a molecule much more easily attackable
than does the alcoholic group CH,OH. Thus, we see that
glycerol and mannitol are (at least in the animal) by no means
easily oxidised, while, ¢.g., tartaric acid is. The influence of the
carboxyl-group also becomes evident when we compare the
bibasic phthalic acid with the monobasic benzoic acid; the
latter resists while the former is for the greater part burned up.
That in the animal the amido-acids (leucin, glycocoll, tyrosin)
undergo combustion with especial facility, yielding thereby urea
was demonstrated by Nencki and Schulizen, as early as 1872, and
later on by Knuteriem, Schmiedeberg, Weiske, and Lewinsky, in
experiments with asparagin. Nencki declared, therefore, the amido-
compounds to be the forerunners of urea.”) This view, which does
not assume a direct oxidation of dissolved proteids, but a pre-
vious splitting into a group of well known amido-acids, is very
well supported by observations of Hofmeister, which prove with
what difficulty peptone, as such, is oxidised in the living animal.
A rabbit of 1.75 kilo. in weight, discharged, after intravenous injec-

(1) Nencki and Sieber have tried to determine the extent of oxidation which
sugar and albumin can undergo if exposed at 36° in alkaline solution to air, and have
found it to be very slight. How little the protoplasm itself participates directly in
the oxidation process may be illustrated by the fact that 95 per cent of the matter
oxidised may be sugar and only 5 per cent proteids; with bees the percentage of
the latter is still less.

(2) The amido-acids from proteids are even more quickly oxidised in the cells
than the carbohydrates, as must be concluded from investigations by Kumagawa,
who nourrished dogs with an excess of lean meat and observed that under this con-
dition also the glycogen of the meat was deposited as fat in the animal (Mittheilun-
gen der medic. Facultat, Tokyo). It is, moreover, of great interest to observe how
the production of urea is influenced by the chemical constitution: negative groups
connected with the nitrogen will prevent the formation of urea, as Nencki has shown
with acetamide, recalling the resistance of hippuric acid. But taurin and sarkosin
are also oxidised with difficulty, reappearing as uramido-compounds in the urine
(Salkowski). Urea must be considered as a synthetical product from carbamic acid
and ammonia (Drechsel and Abel; Nencki and Hahn).

THE ENERGY OF THE LIVING PROTOPLASM. 185

tion of 0.318 g. peptone, over ¢ of it again in the urine, and after
subcutaneous injection about 3. Trypsin, present in minute
quantities in all parts of the body, may gradually form amido-
acids from the circulating protein; also decompositions of an-
other kind may simultaneously take place, whereby sugar is one
of the products, which process can not only occur in the liver
but also in the milt and kidneys (Lefine and Metroz). ‘The great
amount of urea discharged a few hours after a meal may be due
mostly to the amido-acids formed by the pancreatic juice from a
part of the proteids of the food.

The view of Nencki is further supported by observations in
plants. In all cases where the proteids are attacked in plants to
support respiration, a previous formation of amido-acids takes
place, whose residues are finally found in form of asparagin (cf.
Chap. V); a synthetical product corresponding to the urea of
animals.

‘The view here taken as to the cause of respiration, corres-
ponds in its principal feature to the definition Nagel: gives of
oxidising fermentation.’?) This author says: ‘‘ the specific state
of motion in the living protoplasm of the mycoderma cells is
extended simultaneously to the alcohol and to the oxygen
molecules. If, thus, the equilibrium is disturbed to a certain
extent, chemical change takes place by aid of chemical affini-
ties.’ This theory, however, is still imperfect, as it does not
show how the ‘‘ specific state of motion” in the protoplasm is
caused and does not define whether it consists of a molecular or
of an atomic motion.%) The new theory which assumes a
chemical difference between the protceids of the living and those of
the dead protoplasm furnishes the key to the ‘‘ specific state of
motion ;” it infers from physiological facts (cf. Chap. V) the
presence of highly labile atomic groups, viz., aldehyde- and

(1) Asparagin must be defined as the form in which ammonia is stored up in
plants, whether it be formed by decomposition of proteids and amido-compounds or
it be resorbed from tbe soil. Cf. Kinoshita, this Bulletin.

(2) Theorie der Garung, p. 43.

(3) Ndgeli published his ** Theorie der Garung”’ in the year 1879. A few years
later, | often had discussions with him concerning the difference between living and
dead protoplasm, which he had taken for a physical and anatomical one, Later on,
however, he agreed that there must exist a chemical difference.

186 | THE ENERGY OF THE LIVING PROTOPLASM.

amido-groups in the molecules of the active proteids, a conclu-
sion supported again by toxicological observations (cf. Chap.
III), and by the study of the active reserve albumen in plants (cf.
Chap. IV). The actions of this peculiar energy, so well adapted

for transmutation into chemical work, were characterised in
Chap. VI.

It remains therefore only to explain how the intense and
ceaseless oscillations going on in the protoplasm can excite the
atoms in the sugar or fat) to such energetic motions that a
combination with oxygen will result. In this regard it will
suffice to point to the action of heat, which, at a certain tempera-
ture, will bring about the combustion of various compounds.)
The increased moleculay motion passes partially into atomic
motion, rendering the atoms very labile, i.c., imparting to them
intense oscillations, whereby the chemical affinities originally
governing the molecules are loosened, leading to an increased
affinity for oxygen. As cohesion is loosened by heat, so the
atomic cohesion, 7.c., the affinities in a molecule, are loosened by
plasmic energy ; in both cases atomic oscillations are inaugurat-
ed leading to combustion, with the main difference that plasmic
energy can accomplish its effect at a much lower temperature than
heat energy. When the wave impacts originating from the labile
atoms in the protoplasm have fallen upon the atoms of the res-
piratory fuel, and have led finally to motions of a certain intensity,
the act of combustion sets in with great vigour, not a previous, but
a simultaneous splitting up of the oxygen molecule taking place :
normal rvesptvalion. But if molecular oxygen is absent, then such
“activified ’ sugar molecules will undergo other changes, with
the production of fat, lactic acid, or alcohol ; these processes are

(1) Fats as such can hardly serve for respiration, not being soluble in aqueous
fluids. It was therefore supposed that a previous saponification is necessary. It is
however, much more probable that a conversion of the neutral fats into lecithin takes
place, which swells up easily in water and is even a little soluble in it. Cf. O. Loew,
On the physiological functions of phosphoric acid, Biol. C. 11,270.

(2) A previous “ activifying ’’ of the oxygen is here never noticed, for the small
amount of ozone formed by rapid combustion under certain circumstances is only a
by-product. Cf. O. Loew, On the formation of ozone in rapid combustion, American
Journal of Science, Vol. 49.

THE ENERGY OF THE LIVING PROTOPLASM. 187

accompanied by a development of carbon dioxide, and have
received the name : intramolecular respiration.”

If the amount of respiratory fuel decreases, the intensity of
respiration will diminish also, and finally, the active proteids of
the living protoplasm will, on account of their lability, themselves
take up oxygen. And ifina cell a small amount of protoplasm
has thus been changed, the equilibrium of the entire tectonic will
be disturbed and a rapid chemical change will follow the contrac-
tion : death by starvation. The protoplasm, however, as if endow-
ed with intelligence, understands how to avoid this dangerous
result. It absorbs food and instead of itself taking up oxygen,
throws it upon the ‘‘activified’’ molecules of food and thus
derives the greatest profit to itself, the liberated energy being
utilisd for various vital functions. A great part assumes the
form of heat and is dissipated, another part that of mechanical
activity, another, again, serves to increase the original plasmic
energy and leads thus to the most wonderful chemical perform-
ances.

But the increased state of lability will lead in turn to an
increased respiration, and this, again, to an increased lability.
Thus the temperature would continuously rise also, and another
dangerous point would be reached: death by heat, at 45-50°C.
The increased motions of the labile atoms would facilitate the
reaggregation into stable position, the passage into passive
proteids by the action of the labile groups upon each other;
we may define this death as caused by an intramolecular self-
poisoning. However, there exist, again, conditions to prevent
this result, heat being lost by conduction, by radiation, and,
further, by evaporation of water. Higher animals also enjoy the
benefit of regulating contrivances in the nervous system. It is,
nevertheless, admirable to see how close the temperature of
birds is kept to that dangerous degree which, like an abyss,
separates life from death. And still more remarkable are the

(1) The view of several authors that the so-called intramolecular respiration is
the primary cause of normal respiration has been refuted as erroneous by Sachs,
Diakonow, Godlewski, and others. In most organisms absence of oxygen acts
detrimentally, but in cold-blooded animals and plants much more slowly than in

warm-blooded animals, while anaérobic microbes and yeasts can do without it
altogether,

188 THE ENERGY OF THE LIVING PROTOPLASM.

few exceptions to the general rule, as observed in certain alge and
bacteria, which can remain alive even at temperatures above
go® C.

If we now consider again the general teaching that ‘‘the
potential energy of the food yields the kinetic energies of the
organism,’ we must keep in mind that there is originally present
a certain energy in the living matter, which liberates that energy
by combustion, and that the original energy being thus intensified
leads to the vital functions in the various organs and organisms.

On the Reserve Protein in Plants. Il,
BY
G. Daikuhara, Nogakushi.

I have given the results of my investigation on the occur-
rence of active albumen in a large number of plants in Vol. IT.
No. 2 of the Bulletin of this College. As all the objects then
examined were collected in spring, it seemed to me of interest to
examine them during the fall. We know that an extensive
transportation of nutritive materials takes place from the leaves
to the stem, roots, and bulbs. I supposed therefore that leaves
found rich in active albumen in spring might not show this kind
of reserve material in autumn. On the other hand, it seemed
also possible that plants of rapid growth and yielding none of
the reactions of active albumen in summer might yet accumulate
some active albumen late in autumn when development stops.

I wanted also to test the leaves of evergreen plants in the
fall, as well as a number of fruits. Accordingly, all the objects
here mentioned were examined during the months of October,
November, and December. The general result of this examina-
tion has been that objects not showing any active albumen as
reserve-material in spring also show none in autumn ;") and that
most objects yielding a positive result in spring, yield the same
in autumn, although usually to a much less extent.

I have also repeatedly examined leaves partially dead, as
shown by their brown and yellow spots, and could never produce
proteosomes with caffeine in the brown parts, but always in the
still healthy and green cells, even at points very close to the
dead portions. In a few cases I could see, without caffeine
treatment, bright globules in the cells, which in some in-

(1) As examples of those which give no reaction in spring, and do give one in
autumn, I will mention the leaves of Deutzia Sieboldiana, Diospyros kaki, and Salix
japonica. It should be mentioned here that sometimes the caffeine reaction does not
set in immediately, especially in the case of thick cell walls, and can then be
detected only after the prolonged action of a saturated caffeine solution.

190 DAIKUBARA 3}

stances proved to be fat.) In one case, however, viz., in the
epidermis of the midribs of the leaves of Osmunthus Aquifolium,
they resembled «impure proteosomes, and were changed and even
partly dissolved by alcohol of 20 per cent.

The frequent occurrence of active albumen in different parts
of the flowery may have an important physiological relation to the
formation of seeds. However, the seeds and fruits investigated
showed active albumen only in the epidermis in most cases.

A phenomenon which resembles, to a certain extent, the
formation of proteosomes is plasmolysis. Th. Bokorny has al-
ready observed (Pringsheims Fahrb. Vol. 20) that by the action of
caffeine both, normal and anomalous plasmolysis, can take place,
whereby the tonoplast can become divided into two or more
parts keeping their globular shape for some time. According to
his theory, these phenomena have to be explained in the same
way as the formation of proteosomes, viz., as being caused by the
separation of a certain amount of water of imbibition. It was
therefore a matter of particular interest to me to observe such
phenomena myself in a number of cases: as, for instance, in the
petals of Ipomea hedracea,”) the leaves of Camellia theifera, and
the leaf-veins of Pyrus Toringo.

The two kinds of globular formation may, however, be easily

(1) I would here supplement what I have said (loc. cit.) upon the use of alcohol
and ether to distinguish proteosomes from fat globules. In order to preserve the
globular shape of the former, treatment with 1 p.m, NH3 or with alcohol of 20 9/o is
necessary, before the mixture of alcohol and ether is applied. In certain cases the
ammonia has to be diluted to 4 p.m. because of the gradual solvent action of more
concentrated solution.

The caffeine proteosomes are, in exceptional cases, not solidified into bright
globules, by the action of dilute ammonia, but are partially or, in some cases, (Acer
palmatum) wholly dissolved. This is evidently due to the presence cf a large
amount of impurities (tannin especially) in them. The petals of Pwonia yield
caffeine proteosomes, which are turned by ammonia into hollow globules without any
brightness. There exist evidently many differences between the active albumen of
some species, and that of others, ¢.g., many isomers and polymers are possible.

(2) Accompanied by comparatively few proteosmes, and only observed in the
lower colourless part, not in the upper coloured part of the petals, after treatment
with caffeine.

Anomalous plasmolysis by 0.5 °/o caffeine solution can also be sometimes ob-
served in Spirogyra, after it has been cultivated, for some time, in a 1-5 per mille
nutrient fluid.

ON THE RESERVE PROTEIN IN PLANTS. II. Igt

distinguished by treatment with 1 p.m. ammonia, by which the
forms of true proteosomes are not changed,” while the tonoplast
loses its globular shape.)

Another reagent for distinguishing the two kinds of globules
is iodine solution, which produces vacuoles in the true proteo-
somes, while it only causes the tonoplasts to shrivel up.

The results of my examinations are shown in the following
tables:

RANUNCULACEZE.
SPECIES. Opjects ACTIVE REMARKS.
TESTED. ALBUMEN,
Peonia albiflora, Pall. Old leaves Much Even present in those
living cells that
Anemone japonica, Sieb.)| , were near brown
et Zucc.) Blowers None dead portions of
the leaves.
” ” Leaves None
BERBERIDEZ.
Nandina domestica, Thunb. | Old leaves Present
ms + Epid. of fruit Moderate
TERNSTRGMIACEX.
ror 6 (Epid. of old 5 Leaves suffering from
Camellia japonica, L. | leaves | Little aihiatcar also gave
the reaction.
0 a Midribs Very much
” » Epid. of fruit 7 *

Camellia Sasanqua, Thunb. | Epid. of fruit | Very much

” ” ” Leaves ” ”
” » » Flowers an a
Camellia theifera, Griff. Epid. of fruit | Much Small fat globules
present.
” i Flowers Moderate

Ternstreemia —_ japonica, )

Peas Loaves Moderate Also present in the

reddish portion of
| the leaves.

(1) They merely solidiy and seem to shrink a little.

(2) In some cases, however, it is well, after the ammoniacal treatment, to apply
10 O/o acetic acid under the microscope, which does not affect the solidified proteo-
somes, but destroys the globular shape of the tonoplast. I applied this treatment in
the case of the petals of Ipomaa hedracea.

DAIKUBARA 3

MALVACE.
aan OBJECTS ACTIVE
SPECIES. ESTED. a REMARKS.
Hibiscus syriacus, L. Flower None Starch present.
Hibiscus rosa-sinensis, L. Flower None
” ” Leaves None
GERANIACE.
Oxalis rosea, Jacq. Flower None Strong acid reaction.
” ” Leaves None
CELASTRINEA.

Celastrus articulatus, Thunb.

Contains
drops.

many fat

E ; No decisive
Epid. of fruit { TORCLOn \

SAPINDACE#.

Acer palmatum, Thunb. Epid. of leaves | Little
3 of Epid. of veins | Very much
LEGUMINOS#.
Phaseolus radiatus, L. Leaves None Much soluble passive
albumen.
Wistaria chinensis, Sieb. No passive albumen
et duces} acs None in solution.

ON THE RESERVE PROTEIN IN PLANTS. II. 193
ROSACEA,
OBJECTS ACTIVE f
= PEA TESTED. ALBUMEN. ESS
Prunus persica, Benth et\|fOld yellow Very little in the
Hey { leaves GSO: epidermal cells and
moderate qt. in the
Prunus Pseudocerasus, Lindl {Ore as Present pallisade cells.
eaves
5p ep August flower | Little
Prunus Mume, Sieb. et\|fOld yellow Only in the pallisade
Deel leaves } BeGoene cells.
Pyrus Toringo, Sieb. Young leaves | Little
33 ap Veins of leaves | Very much In some cells there
was produced ano-
7 i Epid. of fruit | None malous plasmoly-
sis.
Rosa laevigata, Mich. Epid. of fruit | None
Rosa indica, L. Leaves None
¥5 5 Flower Very much
Photinia glabra, Thunb. Epid. of leaves | None
Spongy tissue :
ve my { of leaves j SHER
SAXIFRAGE.
Deutzia crenata, Sieb. et | No passive albumen
yes} EEE Very much in solution.
Astilbe chinensis, Maxim. Leaves None
LYTHRARIE.
Lagerstreemia indica, L, Flower Very much
” ” Epid. of fruit | Very much
Punica Granatum, L. Epid. of fruit | Very much

194 DAIKUBARA 3

BEGONIACE.
OBJECTS ACTIVE
SPECIES. Pa Coe REMARKS.
Begonia Evansiana, Andr. | Flower None
Tenves No decisive
» 20 a2 reaction
ARALIACEZE.
Aralia cordata, Thunb. Leaves Little

3 Epid. of fruit | Little

9

CAPRIFOLIACE.
|

Sambucus racemosa, L. ; Many fat globules
var. Sieboldiana, mia Teayes EES: present also.
EBENACE.

Diospyros kaki, L. | Old leaves | Much | No passive albumen.
OLEACE.

Osmunthus Aquifolium, :

Benth et ee Flower Moderate
+ i Epid. of veins | Little
” ” Leaves None
CONVOLVULACE.

Ipomeea hedracea, L. Leaves None Even in midribs no
caffeine reaction
during flowering.

” 5 Petals of flower} Present Only in large cells
near spiral vessels.
Anomalous plasmo-
lysis also produced.
” ” Siamiens et Present Also anomalous plas-
flower :
molysis produced,
by caffeine.

—e. iia

ON THE RESERVE PROTEIN IN PLANTS. II. 195

SOLANACE.
SPECIES. Osjects ACTIVE REMARKS.
TESTED. ALBUMEN.
Solanum Melongena, L. Flower None
rr is Leaves None Little passive albu-
men in solution.
Pe Ar Epid. of fruit | None
Nicotiana ‘Tabacum, L. Leaves None
” oh Flowers None
nr an Epid. of seeds | None
POLYGONACE®,
Polygonum ieee Blowers Very much
our.
5 7 Leaves None
CUPULIFER.
Quercus glandulifera, BI. Leaves Little
” ” Midribs Much
SALICINEA.
Salix japonica, Thunb. Leaves Very much |
SCITAMINE#.

Canna indica, L. | Flower | Much present

On the Consumption of Asparagine in the Nutrition of Plants.
BY

Y. Kinoshita, Nogakushi.

The fact that asparagine is formed whenever proteids undergo
decomposition in plants has been repeatedly made the subject
of close investigation by various authors, but much less atten-
tion has been hitherto paid to the reverse process—the
regeneration of proteids from asparagine. C. O. Maiiller™) has
asserted that this regeneration can only take place during the
process of assimilation in green leaves, and that the action of
light and the status nascens of carbohydrates are essential. As
this statement did not appear to me to be well-founded, quanti-
tative investigations being wanting, I undertook a series of
experiments in order to see whether this process might not
proceed in the dark.

If we look upon the growing root from the physiological
point of view, we must hold it highly probable that the cells of
the root, although always deprived of light, are capable of
forming the proteids necessary for its growth and development
from suitable sources, such as sugar, nitrates, and sulphates, or
sugar, asparagine, and sulphates. It is a well known fact that
mould fungi can form their proteids in this way, in complete
darkness, and that certain fungi, especially bacteria, grow even
much better in the dark than in day-light; it may, therefore, be
surmised that the formation of proteids and protoplasm may also
proceed better in darkness.

The relative amount of glucose, or other suitable material,
seemed to me the most decisive factor in the transformation of
asparagine into proteids. I, therefore, selected shoots of soya-
beans, which are rich in asparagine, and tried to nourish them
with organic materials, repeatedly making microscopical tests,
in the usual way, with alcohol, as described by Borodin, Pfeffer,

(t) Ein Beitrag zur Kenntniss d. Eiweissbildung in der Pflanze. Landw.
Versuchst. Bd. 33, p. 11.

CONSUMPTION OF ASPARAGINE IN NUTRITION. 197

and others, and finally determining the amount of asparagine
still present, by crystallisation, according to Z. Schulze’s method.“
Either methyl alcohol, glycerol, or glucose was used as organic
nutrient, in solution along with calcium sulphate.?) Every
seventh or eighth day, this solution was replaced for a day by
one containing 0.5 per mille each of the two potassium phos-
phates, and of hydrated magnesium sulphate, so as to furnish
the necessary mineral matters. The cotyledons were cut off at
the beginning of the experiment, in order to prevent further
formation of asparagine by decomposition of the reserve proteids.
A control experiment was made at the same time with soya
shoots kept in water, in which an asparagine determination was
made just before the other shoots were placed in their nutrient
solutions, and again in others at the time when the shoots under
investigation were analysed.

The soya-beans were soaked in water on March 7th, sown
on moist saw-dust next day, and kept in the dark. In four days,
at rather low. temperatures, the seeds had germinated and the
roots had reached the length of 2-3 cm. At this time I tested
the tips of several roots microscopically for asparagine and found
only doubtful traces; but when the roots had become 6 cm. long
the presence of a moderate quantity was easily recognised. At
this time the young plants were placed in water on a wire net.

After the length of the entire plants had reached 20-27 cm.
and the stem and the root had been found to be rich in asparagine,
by microscopical tests, a portion of the shoots was placed on April
I., (a) in ar % solution of methyl alcohol mixed with 4 of its
volume of saturated gypsum solution, (6) in glycerol") solution
of r %, with gypsum, and (c) in glucose solution, the cotyledons
of all the shoots having now been removed.

Whenever the solutions became turbid from bacterial growth,
they were renewed at once. When the hypocotylous part
of the stem had reached about 30 cm. in length, growth seemed

(1) Landw, Jahrbiich., 1888, p. 688 and 701; also 1880, p. 14.

(2) In some trials I made use of sodium acetate and tartrate, but not with
satisfactory results, perhaps because the conditions were not favourable.

(3) Preliminary experiments had convinced me that a solution of 10 °/o or even
of 5 °/o glycerol is not adapted for the further development of the plant. Indeed, in
the 10 °/o solution the shoots died after 2 days.

198 KINOSHITA ; CONSUMPTION OF

to stop in it, while the growth of the shoots above the cotyle-
dons was now more marked than before. It may be mentioned
that most of the leaves of the shoots cultivated in glycerol
solution were somewhat larger than those grown in methyl
alcohol. Microscopical examination exhibited now a very
great difference between the amount of asparagine present in
the control shoots, and that present in the other cases. Direct
tests for the presence of dissolved reserve albumen, made upon an
aqueous extract by addition to it of nitric acid, showed that there
was none present in the control case and much present in the
shoots cultivated in sugar and glycerol. We see, therefore, that
the decrease of asparagine is coincident with an increase of the
dissolved proteids. Microscopical tests made it further highly
probable that the amount of other amido-products was con-
tinuously decreasing, while tests for sugar with Fehling’s
solution revealed its presence in the shoots grown in glycerol, but
neither in those grown in methyl alcohol nor in the control case.

Soon afterwards, on April 27th, a final measurement of
dimensions, and a quantitative determination of asparagine were
made. The stem without the hypocotylous part had a length of
4-14 cm., In the control case No. 2; a length, of 11-19 cm.
in glycerol, and of 8-1g cm. in methyl alcohol.

The quantity of asparagine was as follows:

Asparagine
Date of Dry matter Asparagine per cent in dry

determination. in grammes. in grammes. matter.

Control shoots No. r. April 1st. 3-966 0.853 21.5
An No. 2. pe 27 tie 2.948 0°847 28.7

” ” No. 3. 99 3-611 0.906 24.0
Shoots in methyl alcohol ,, ,, 2.698 0.511 18.9
ee glycerol) ,, ,, 4.590 0.629 13-7

(1) For determining the amount of asparagine in those shoots which had been
kept in the dilute glucose solution, the material was not sufficient, but I micros-
copically examined the shoots for asparagine on the 8th May, (when some of the
leaves showed brownish spots, indicating a gradual decay), and found a not
inconsiderable amount of it still present. I do not doubt that if we could
introduce more concentrated sugar solution into the cells of the shoots, the shoots
would continue to grow in the dark until all the asparagine had been transformed into
proteids or protoplasm, provided the necessary mineral salts had been also
introduced.

ASPARAGINE NUTRITION OF PLANTS. 199

The determination of asparagine in the control shoot No. 1,
was made after the removal of the cotyledons (April 1), when
the experiment proper commenced. If we compare this result
with that yielded by the control shoot No. 2, we find an increase
of asparagine in percentage of dry matter, due probably to the
gradual conversion of other amido-compounds into asparagine.)
The fact that in the control case No. 3, where the cotyledons
had not been removed, a smaller percentage of asparagine was
found than in No. 2, may probably be due to the galactans and
other carbohydrates gradually becoming soluble and getting
consumed in support of respiration, thus protecting proteids as
well as amido-compounds from further changes, and retarding
the production of asparagine.

The principal conclusions which we can draw from the
results obtained are:

(1). Glycerol and methyl alcohol supplied by the roots, can
not only hinder the production of asparagine in the shoots, but
are also capable of diminishing the amount already formed.

(2). Glycerol is much more effective than methyl alcohol.
It also forms sugar in the cells.

(3). Since shoots have been found to grow better in solu-
tions of methyl alcohol and glycerol than in water, and also to
show the presence of dissolved proteids by the nitric acid test,
it is safe to assume an increasing protein production in the shoots
thus nourished; in other words methyl alcohol, as well as gly-
cerol, can serve for the regeneration of proteids from asparagine,
and as this process can go onin ferfect darkness, light must be
denied to have any direct action in supporting it.)

(1) Compare O. Loew. This Bulletin, Vol. II. No. 2.

(2) This formation of sugar is in accordance with the observations of Laurent,
Ar. Meyer, and Th. Bokoyny. ‘The latter author has also observed starch formation
from methyl alcohol under the influence of daylight.

(3) It is however indirectly of great importance, because it yields the necessary
carbohydrates; for the more sugar there is present in a cell, the quicker will
asparagine be transformed into proteids.

On the Assimilation of Nitrogen from Nitrates and
Ammonium Salts by Phaenogams,
BY

Y. Kinoshita, Nogakushi.

There exist many observations on the development of plants,
as to whether nitrogen is supplied in the form of nitrates or in
that of ammonium salts. One would naturally expect that
ammonium salts would be the better source of nitrogen, because
nitrates would have to undergo reduction to ammonia before
assimilation proper (formation of proteids) could take place, thus
causing not only loss of time, but also the waste of such organic
material (most probably glucose) as serves for this reduction.
But, contrary to expectation, nitrates have been found in many
cases to act more favourably than ammonium salts. This result
may be due to the noxious qualities which ammonium salts
have in a higher concentration than that immediately needed
in the cells. Nitrates can be stored up in roots and stems,
but ammonium salts cannot. The question, therefore, presents
itself, as to the form in which the nitrogen of ammonium salts is
stored up, when these salts are absorbed in greater measure than
that required for the immediate wants of the plant.

Asparagine has been frequently found in the roots and bulbs
of many plants, but it has not been positively ascertained yet,
whether this compound is in these cases a decomposition
product of proteids, as it is in germination, and in the starva-
tion of plants kept in the dark. Some authors have assumed
that this asparagine is formed in the leaves by the action
of nitrates upon carbohydrates, and is then transported to
the roots, while others have supposed its synthetical formation
in the roots, leaving entirely unsettled whether nitrates,
or ammonium salts, or both, equally well contribute to its
formation.

NITROGEN FROM NITRATES AND AMMONIA. 201

I have, therefore, made several experiments to decide under
what conditions the formation of asparagine is brought about,
and whether also its formation takes place in the dark, that
is, without the reducing action exerted by light in leaves. For
these experiments, I selected such plants as are rich in starch
and which form, therefore, under ordinary conditions, but very
little asparagine by the decomposition of proteids.

Barley seeds were distributed in three pots, filled with
moist sand, and kept in the dark. After sixteen days the height
of the young plants was, on an average 20 cm., but their tips
began to dry up gradually. At this time, I subjected the plants of
one pot to analysis, while the second pot was treated with a 1 %
solution of ammonium chloride, and the third with an equivalent
quantity of sodium nitrate. Of each of these solutions, 0.5 litre
was administered at three different times. After one week, the
growth was found rather insignificant, and the drying at the tips
had extended.

After the sand had been carefully separated by washing, the
whole plants were analysed. Not a trace of ammonia could be
discovered in them, in spite of their treatment with ammonium
chloride. The total nitrogen was determined by Kjeldahl’s
method, the protein nitrogen by that of Stuézer, and the asparagine
by that of Sachsse.

The result was as follows :—

Date 15th May 22nd May

Plants in water, amm. chlor. sod. nitr.
Total nitrogen 3.512 4.436 4-925
Protein nitrogen 2.704 2.126 2.006
Nitrogen in asparagine”) 0.656 2027 0.977

A very great difference is here seen in the quantity of
asparagine, its production being favoured by ammonium salts.

In a second experiment, young maize of nearly 4o cm. in
height was placed with its roots in solutions of sodium nitrate
and of ammonium nitrate, both containing 0.1 % of nitrogen,
while the control plants were placed in distilled water. After

(1) No especial attention was paid here to the other amido-compounds, which
evidently must have been present in all three cases.

202 KINOSHITA ; NITROGEN ASSIMILATION.

four days’ standing in diffused day-light, the whole plants were
subjected to nitrogen determination, with the following result :—

Plants in water, amm. nitr. sod. nitr.
Total nitrogen 4.13 4.23 4.15
Nitrogen in asparagine 0.38 0.73 0.24

From this it is evident that ammonium salts are transform-
ed into asparagine, while nitrates can be stored up as such. The
difference would have been found still greater in the latter case
had only roots and stems been tested, and I think it safe to
assert that asparagine is the form in which the excess of nitrogen,
originally in the ammonium salts, is stored up in the plants,
although I regard these investigations merely as a preliminary
step to a longer series I intend to carry out in order to decide
whether my conclusion can be substantiated.

|
l]
|
{
|

On the Presence of Asparagine in the Root of Nelumbo nucifera.
BY
Y. Kinoshita, Nogakushi.

The root of Nelumbo nucifera is used in this country in the
boiled condition as food, and is rich in starch. Its nutritive
quality may be judged from the following analysis by Kellner :

WPAECT@ kegs SUA S/8 scot cotgycan = toass A vaey sot beat ORrOAe %
In too pts of dry matter:

Crude HrOveiny Fao, ee ees ooh nee | ce eee FAS

He Aeeu reed Ae) Susecie Foods a maoa liseae ty 850) cesie pedo Be UA

GC UMra a punt aan es och. cong cash, cumple POLO

Non-nitropenous extract; starch ete... =... <«.. 78.59

Ash (free from carbon and carbon dioxide) ... 5.03

I suspected that a certain amount of nitrogen might be
present in the form of amides as in potatoes and beets, and
actually found the presence of a not inconsiderable quantity of
asparagine. This substance is evidently closely connected with
the formation of proteids, and is therefore of high physiological
interest.

It has been frequently found in germinating seeds, which
contain the more of it the poorer they are in carbohydrates. It
is also present in many buds during their development, but its
presence in roots has less frequently been demonstrated. The
root of Althea contains 2%, that of Glycirrhiza (Plisson) 0.8 %,
that of Scorzonera (Gorup) 0.6%, and potatoes 3 % (E. Schulze).
It was also found in the root of Symphytum and in that of Lactuca.

In order to obtain asparagine from the root of Nelumbo
nucifera, | proceeded as follows :—

The fresh root was reduced to a fine pulp and pressed, the
residue was mixed with a little water and pressed once more.
The filtered liquid was evaporated and the residue was extracted
with hot alcohol of 60 %. Upon cooling an abundant crop
of crystals was obtained while the mother liquor yielded more

204 KINOSHITA ; ASPARAGINE IN NELUMBO.

crystals, on treating it again with hot alcohol and evaporating.
An approximate determination showed that this root contains
asparagine amounting to nearly 2 % of the dry matter.

The purified crystals showed not only the same crystalline
forms as asparagine but also yielded up one molecule of water
when dried at 100°, while the nitrogen determination according
to the method of Kjeldahl also agreed with that for asparagine.
For the determination of nitrogen, I took 0.3 gr. of the substance
dried at roo° and obtained 0.06232 gr. nitrogen, that is 20.78 %.

Calculated. Observed.
Water of crystallisation’... ... 7 12ire eG. rea
Nitrogen, .i-g cita sd Weta, <sed acer tee 2002) Pomme enemas

The solubility in water nearly agrees with the number
given by Guarescht, thus:

I part of asparagine dissolves I part of the substance dis-
in 55.86 pts of water at 10.5° | solved in 56.72 pts of water at
(Guarescht). a FF

On the Occurrence of Two Kinds of Mannan in the
Root of Conophaltlus konyaku,

BY
Y. Kinoshita, Nogakushi.

The root of Conophallus konyaku has been recently in-
vestigated by Mr. C. Tsxyt, who found that it contains a large
amount of an anhydride of mannose, but it was not shown by him
whether only one or several polyanhydrides are present. It
has been proved, however, that there exist both soluble and
insoluble mannans; to the former belongs, for instance, the
mannan discovered in yeast, to the latter that found in the nuts
of Phytelephas. The fact that an aqueous decoction of the
konyaku-root is so slimy that it serves in this country for
technical purposes, as for pasting clothes, etc., made it very
probable that some of the mannan is present in the soluble
form.

Special experiments have convinced me that, by treatment
of the konyaku-root with diluted sulphuric acid, neither glucose
nor galactose, xylose nor arabinose is formed. The sugar
appears to be mannose and nothing else. I could neither
obtain mucic acid by its oxidation with nitric acid nor the red
pentose reaction with phloroglucin and hydrochloric acid, nor did
the filtrate from the mannose-phenyl-hydrazone (prepared in
concentrated solution) yield any notable amount of an osazone.

I extracted a portion of the finely ground root repeatedly
with boiling water, until the extract had no longer any slimy
character. The residue yielded mannose, upon boiling with
dilute sulphuric acid, as was ascertained, beyond doubt, by its
phenyl-hydrazone. The slimy extract of the root was mixed
with alcohol, which formed a copious, almost white, flocculent
precipitate, that was filtered off and washed with dilute

alcohol. Its solution in water was so opalescent that no test

(t) This Bull. Vol. II. No. 2.

206 KINOSHITA ; MANNANS IN C. KONYAKU.

_could be made in regard to its rotatory action upon polarised
light. Upon drying at 100°, it lost its solubility in boiling water
entirely. Upon boiling with 4% sulphuric acid for several
hours mannose was also obtained from it. The property of losing
its solubility upon drying proves that this mannan is different
from that separated from yeast by Salkowskz,“) with which it
agrees, however, in the following respects :—

Basic lead acetate—no precipitate; basic lead acetate and
ammonia—thick precipitate; ferric chloride and ammonia—
gelatinous precipitate; copper sulphate and sodium hydroxide—
thick blue precipitate; and the same by Fehling’s solution.

It seemed to me of interest to determine whether the diastase
of malt, invertase, or emulsin would have saccharifying power
over this slimy mannan of konyaku, and this I tried and found
not to be the case; its slimy character remained, although I
digested it with the enzymes mentioned for 5-6 hours at 40°—
60°, and no sugar was formed.

That this mannan is also digested with much more difficulty
than starch, has been shown by Prof. Osawa’s experiments
on dogs. An enzyme endowed with saccharifying power for
mannan must, however, exist in the konyaku-root, and I intend
to try to isolate it in the season when the root is in the state
of developing shoots.

(1) Ber. d. chem. Ges., 1894. p. 499.

Note on the Chemical Composition of some Mucilages,
BY,

K. Yoshimura, No;ahkushi.

The mucilages or saccharo-colloids, hitherto analysed,
have been found to consist, in most cases, of saccharo-polyan-
hydrides of either glucose, galactose, mannose, or arabinose.
Only in one case was the mucilage shown to consist of a mucin
(Ishi, Vol. II. No. 2 of this Bulletin).

Although such compounds are widely distributed in the
vegetable kingdom, they have been investigated but in a very
limited number of cases. As it is of physiological interest to
know the composition of mucilages in as many plants as possible,
I have examined those of the following species :

1. Sterculia platantfolia (young shoots)."?

2. Colocasia antiquorum (tuberous roots).()

3. Opuntia (fleshy stem).

4. Vitis pentaphylla (stems and leaves).

5. Cnothera Faquini (stems and leaves).

6. Kadzura japonica (young leaves and stems).

The concentrated slimy extracts were precipitated with
strong alcohol and the precipitates, after having been washed
with alcohol, were boiled with sulphuric acid of 2-4 % for 2-5
hours, the liquid neutralised with barium carbonate, and the
filtrate evaporated to a syrup.

A portion of this syrup was evaporated with nitric acid to
observe whether mucic acid was formed.

Another portion was mixed with a cold concentrated solu-
tion of acetate of phenylhydrazine to observe whether mannose-
phenylhydrazone was formed.

(1) The mucilage of Sterculia platanifolia, as well as that of Kadzura japonica,
finds technical application in this country, being used for sizing paper, etc.

(2) The tuberous rootstock of Colocasia antiquorum serves as a valuable food in
this country, and is hence cultivated to a large extent.

208 YOSHIMURA ; COMPOSITION OF MUCILAGES.

Another portion was examined with phloroglucin and
hydrochloric acid for the presence of pentoses. And, finally, the
osazones were made in the usual manner and, after purification
by recrystallization from dilute alcohol, their melting points were
determined. In this manner, I was enabled to come to the con-
clusion that the mucilage of Sterculia platantfolia consists of a
mixture of araban with some galactan, and that of Colocasia
antiquoruim, since it gave neither mucic acid nor the pentose or
mannose reaction, but an osazone which was proved to be
identical with phenylglucosazone, consists probably only of a
polyanhydride of d-glucose.

The mucilage of Vitis pentaphylla, as well as that of Opuntia,
consists principally of galactan, while those of CSnothera Faquini
and of Kadzura japonica contain galactan and araban.

The Preparation and Chemical Composition of Tofu.
BY

M. Inouye, Nogakushi.

Wherever rice forms the principal food for man, as in Japan
and China, an addition of some other food richer in proteids is
necessary to make up the deficiency of proteids in rice.”) The
inhabitants of this country on the sea coast supply this want
by the use of marine animals, while the inland people subsist
on the seeds of various leguminous plants, especially the soya
bean, beef and other kinds of meat having only come into use
in recent times. ‘The soya bean is considerably richer in
proteids than rice, as may be judged from the following
analyses :—

RIcE.(?) SOYA BEAN.
I 2 3 4 5
Crude sprotemi A.) sj) 10.82 33.69 31.21 34-92 33-36 42.05
IPB tog 6p soo ote sou 2.78 17.87 18.29 15-53 21.86 | 20.46
CGO s, co oo vo 1.12 12.69 T2373 12.81 -- 4-53
SAM Noc! G6) ab- oo. po 84.22 240 Best 3-53 —- —
IAS Go RE NOE) bbe "De 1.06 5-39 5.63 5:97 5-35 4-19
Other organic matters .. _- 21.01 28.09 26.53 34.18 28.82

(x) Although the proteids of rice are better digested than those of barley, as
Rintaro Mori has shown, the exclusive use of rice cannot be recommended.

(2) The sample of rice and that of soya bean No. 5 of the above table, were
analysed by O. Kellner and were products of Japan (Bulletin vol. I, No. 2 and 6).
The sample of soya bean, No. 1, was from China, No. 2 from Hungary, No. 3 from
France, and analysed by Pellet (Chem. C. B., 1880, p. 410). The sample No. 4 was
analysed by Giissmann (Chem, C. B., 1890, p. 133). .

210 M. INOUYE,

According to the results obtained by Pellet, only about 5 % of
the entire nitrogen is non-albuminoid, while in the Japanese
soya bean, according to the analysis of O. Kellner and Kuro-
shima, it amounts to 7.1 %.

Meissl and Bécker (Chem. C. B., 1883, p. 619) found that
there are no gluten-proteids contained in the soya bean and only
very small quantities of amido-compounds. They extracted the
beans both with dilute potash and also with 1% solution of
sodium chloride, and found in both cases one and the same pro-
teid, which proved to be a casein closely related to milk-casein
and to the legumins of other leguminous plants.

I have confirmed the absence of starch noticed by Kellner
in the Japanese soya bean, by repeated tests with iodine, while
observations made in Europe have shown the presence of a very
small quantity of starch granules. The numbers given above by
Pellet for starch include also dextrin-like bodies, which were
closely examined by &. Schulze, who found them to consist of
two kinds of galactans. He also discovered in them some cane
sugar and determined the amount of lecithin to be 1.64 %.

Stingl and Morawskt (Chem. C. B., 1886, p. 724) found a
very active diastatic enzyme, possessed by the soya bean much
more largely than by many other leguminous seeds. The diges-
tibility of the bean was determined by Giissman. He found that
go % of the protein present is digestible, 89.8 % of the fat, and
14.5 % of the crude fibre.

The efforts to prepare an easily digestible food from soya
beans led to the preparation of miso and natto, two kinds of
vegetable cheese, which were investigated some time ago in the
laboratory of this College.)

But the most interesting preparation is tofu, which consists
principally of the protein-matter of the soya bean, and which,
according to the investigation of Prof. Osawa in Tokyo, is as
easily digestible as beef. This preparation is freshly made every
day, and sold in form of tablets about 10 c.m. broad, 2 c.m.
thick, and 25 c.m. long, is of snow-white appearance, and of the
consistency and taste of freshly precipitated casein of milk, but

(1) On the preparation of miso, by O. Kellner, this Bulletin, Vol. I, No.6. On
natto, by Yabe; Bulletin Vol. II., No. 2.

THE PREPARATION AND CHEMICAL COMPOSITION OF TOFU. 2I1I

as there is no trace of bacterial action connected with its
preparation, the name vegetable cheese, is certainly not
justified. The constituents, as determined by Kellner, are as
follows :—

a a toe age te ee ela, 35a, ‘sete pus” § OG. 20
PMUORMER OTS: 3. Mute, CCN AEM tsy piisin, “ssuteMysnsees (4.07
MonemiIKOSenous, EXtEACtts.. i.) - sss .sse> wae A695
Ash es | ot Ea De crn sip ycoutly tee O24

Tofu is also sold in another form, called kori-dofu. It is
prepared by exposing the fresh tofu tablets to the action of
frost, under which they shrink considerably, lose water, and
become more compact. While fresh tofu contains, on an aver-
age, 89.02 % of water, kori-dofu contains only 15.32 %, in the
air dry condition.

The analysis of kori-dofu gave me the following results :—

Rsetbetea © si, oe a ee wo. hed, “Mtes ellsek >) TRAD 9%
PE UMNNOS ie Men. fn! aaemes, “Aida %
Habeanamlectt hint) seme er. waae eames 2305’ OF

Non-nitrogenaus extract® “..% ic) <<. ae F505 %
WeMMlOsers.. lacs ienaMedceee ce Sissy + suet | Sdn Aie. 96
chipeee se AUG era e rao ws; | SA) Bk 1 MBLOS: Of

The manufacture of tofu") is conducted as follows :—

The beans are first soaked for about twelve hours in water
and then crushed between two mill-stones until a uniform pulpy
mass is obtained. ‘This is then boiled with about three times its
quantity of water for about one hour, whereupon it is filtered
through cloth. This liquid is white and opaque, exactly like
cow’s milk; while the smell and taste remind one of fresh
malt. Upon standing, very fine particles separate on the surface
which, under the microscope, can easily be recognised to be small
globules of fat. Its reaction is neutral or very slightly acid, but
after standing for several days it gives a strong acid reaction, ow-

(1) Tofu is manufactured only ona small scale, by people who sell it in their
own shops.

(2) Even continued boiling will not bring on coagulation of the milky liquid,
but the addition of an equal volume of alcohol yields a flocculent precipitate.

212 M. INOUYE;

ing to the formation of lactic acid, when the separation of casein
takes place exactly as in the souring of milk. The lactic acid
was determined by shaking out the filtrate repeatedly with ether,
evaporating the ether, and boiling the syrupy sour residue with
some water and finely suspended oxide of zinc. The filtrate was
evaporated to a small volume and after a few days the crystal-
lised lactate of zinc was collected, dried, and weighed.

Thus, 100 ce. of the milky liquid, that had been left.for two
weeks in contact with air, yielded 0.13 grm. of lactate of zinc,
corresponding to 0,092 grm. of lactic acid.

I also analysed the fresh milky liquid with the following

results :—
Soya bean milk. Cow’s milk.
Witter cco geceieteeess.> oes. a be ROSES IO EOOROOMEE
Albuminoids FEM Gig anc: Gecele eegsOemre 4.00 %
Fat Lace REESE osha), gee ZaG aes 3.05 %
Fibre’ o.u) Seamer V0 arc OnOR Mere —
Ash) Go 3: each ee +> + Sas Owetans G70
Non-nitrogenous extract, includ-
ing carbohydrates ... «=.» 188 °% =
Milk Stigaraemerrigeee soho —_ 5.00) ve

The fat contained in this liquid as well as in the ¢ofw-tablets
was found to consist partly of lecithin. Tofu dried at 100°
yielded 26.65 % fat and 4.83 er. of this fat yielded, after igniting
with carbonate of soda and nitrate of potash in the usual way,
0.280 grm. of magnesium pyrophosphate, which, multiplied by
the lecithin-factor, 7.2703, corresponds to 2.035 grm. lecithin,
amounting to 11.2 % of dried tofu, leaving for the genuine fat
15.4 %;%) more of the latter, therefore, is left in the refuse than
of the former.

In the manufacture of fofu-tablets from the freshly prepared
milky liquid, about 2 % of concentrated brine as it is obtained as
mother liquor from the preparation of sea salt, is added with
constant stirring, whereupon a flocculent precipitate is soon
formed which is separated by means of a cloth filter, slowly

(1) A portion of this lecithin was probably present in the soya bean as lecith-
albumin; comp. Leo Liebermann, J. B. f. Thierchemie, 1893, p. 32, and E. Schulze,
Chemiker Zeitg. 1894, Nr. 43.

THE PREPARATION AND CHEMICAL COMPOSITION OF TOFU. 213

pressed, and then cut into tabular shape.“ I have tried to
arrive at a satisfactory explanation of the nature of tofu, and have
found that the salt-brine does not act by its chloride of
sodium, but by the calcium and magnesium salts which are
in it; for we can at once obtain the precipitate from the
milky liquid if we add a little calcium nitrate or magnesium
sulphate,” while we can not obtain any separation or precipita-
tion by adding even considerable quantities of sodium chloride
or sodium sulphate.°) However, ammonium sulphate will, as
with cow’s milk, bring on precipitation, if we add 60% of it to
the milky liquid. This precipitate is so voluminous that the
liquid seems to solidify. The peculiar state of the solution of
soya bean casein resembles that of milk; thus, if milk is
dropped upon porous clay, the liquid will be absorbed by the
clay, but the casein and fat will remain on the surface, and the
same is observed with the milk from soya beans.

I have analysed a sample of the salt brine used for tofu mak-
ing and found it to contain, besides chloride of sodium, 27.9 % of
chloride of magnesium and 7.0 % of chloride of calcium. As
casein has decidedly an acid character and is only very little
soluble in water in the free state, it seemed most probable that
the aqueous extract of soya beans contains a sodium or potas-
sium compound of casein®) yielding insoluble calcium and
magnesium compounds, which constitute tofu.

(1) In this way about one fourth of the total amount of proteid in soya beans is
obtained in the tofu.

(2) An excess of magnesium sulphate is to be avoided, as it would redissolve the
precipitate.

(3) Ifthe liquid is warm and saturated with sodium sulphate, a very slow and
imperfect separation may be noticed.

(4) In order to see whether a product similar to Swiss cheese could be obtained
from the crude soya casein or tofu, I infected 50 grm. of fresh tofu with a small dose
of pulverised Swiss cheese, and added ten per cent of common salt to the mixture,
pressed it in cloth, and allowed it to stand in a moist beaker glass for several
months. The product resembled, only to a limited extent, the cheese from milk, but
further experiments with addition of small quantities of milk sugar are intended.

(5) I observed that soya bean casein is easily digested by pepsin solution
acidulated with 0.2 /o hydrochloric acid, without leaving any insoluble residue ;
in this it resembles human casein, which, according to the valuable investigations
of Wroblewski (Berne, 1894) yields, unlike cow’s-casein, no paranuclein. I found

also that peas and horse-beans yield aqueous extracts of similar behaviour to that
of soya beans.

214 M. INOUYE;

I found that, after extracting 100 grms. of fresh tofu with
1% acetic acid, the filtrate contained lime 1.g1 % and magnesia
3.82 % of the dry matter of tofu. A part of this lime and mag-
nesia may have been present as phosphate in the tofu, but another
part must have been in it in combination with the casein, as
also becomes evident from the different behaviour of tofu towards
a diluted solution of disodium phosphate. If tofu is boiled with a
1% solution of this salt, the casein goes into solution and soon
forms an opalescent liquid, while the calcium of the tofw yields
calcium phosphate; if we, however, prepare at first the free casein
from tofu by treatment with very dilute acetic acid and thorough
washing, we find that it will be soluble only in traces in disodium
phosphate even after prolonged boiling, while it is easily soluble
in the carbonate. The free soya bean casein thus obtained will,
after complete removal of the fatty matters by alcohol and ether,
still give an opalescent solution when treated with dilute potash,
showing that the opalescence is due not only to suspended fatty
matters, but also in part to the casein itself. Such a pure
solution of the potassium compound of the casein behaves to-
wards calcium and magnesium salts exactly like the original
liquid from which ¢ofw is made.

The results which we arrive at in regard to the prepara-
tion of tofu is as follows: In the soya beans there are contained
compounds of casein with potassium or sodium which are not
coagulated by boiling, but which yield with the calcium and
magnesium salts of the brine that precipitate of insoluble
calcium and magnesium compounds of the casein, which con-
stitutes tofu. The separation of tofu is therefore not due to
coagulation but simply to precipitation.

The great similarity between concentrated extract of soya
beans and animal milk is also exhibited by the formation of thin
surface films on evaporating it at high temperatures. These
films contain not only the proteids of the liquid but also include
suspended fatty particles and other impurities which are present
in the solution. Such films are prepared in this country by
evaporating the extract of soya bean, and when dried form

(t) Therefore we can safely infer that the solubility of the crude soya bean
casein in hot water is not due to the presence of alkaline phosphates.

THE PREPARATION AND CHEMICAL COMPOSITION OF TOFU. 215

an article of food called yuba. Analyses of the air dry product
gave the following results :—

NIV ENE” one” leet! @ @ga aaa eae 21C0 1 ae A
OKCIASMMI Ne Gon Soe mae (ahs, wie ade, AO! %
EE 5) ot eect y esc, c25: | ute QAOR
Non-nitrogenous compounds .... ... ... =7.65 %
AMEE CA ce mite cree eae so | see 2eO Ze

Carbohydrates. trace.

Note on Nukamiso.
BY

M. Inouye, Nogakushi.

Nukamiso is rice-bran in a state of lactic fermentation, and
is used for softening certain vegetables, such as the fruit of the
egg-plant and the radish, which are rendered palatable and
easily digestible, when left in a large quantity of nuwkamiso for
about 24 hours."

Nukamtso is prepared by mixing rice-bran with about four
times its amount of hot water and adding 6-10 per cent of salt
to the mixture. A small quantity of old nukamiso is added
in order to give the fermentation an early start. Very
frequently also some fish broth is added, whereby the fer-
mentation is considerably enhanced. The mixture acquires
gradually a strong acid taste and peculiar smell, and has to be
stirred from time to time to avoid the growth of mould fungi on
the surface, which would no doubt use up the lactic acid and,
by thus rendering the mass gradually neutral, would allow of the
growth of putrefactive bacteria, which would spoil the product,
more especially if enough salt had not been added. Comparative
experiments with one per cent solution of peptone have shown
that if the quantity of sodium chloride sinks below 5 per cent,
putrefaction can easily set in,’*) while lactic fermentation of a
sugar-peptone-solution is not prevented by even 10 per cent
of sodium chloride, although it is much retarded.

(1) The nukamiso is not eaten, but is carefully washed off from the articles that
have been treated with it.

(2) An addition of 5 per cent sodium chloride to a peptone-solution will retard
the putrefaction to such a degree that after about two weeks the amount of ammonia
formed is only about } that of the control case without sodium chloride. This
amount of ammonia would be still less, if some lactic acid were present, as in nuka-
miso; moreover, 1 may mention that, although an addition of even 8 °/o of sodium
chloride to the diluted peptone-solution will not prevent bacterial growth, putrid
fermentation is very much retared, if not entirely stopped, by that addition, especi-
ally if 0.5 °/o of lactic acid is still present.

NOTE ON NUKAMISO. 217

Microscopical investigation of a sample of nwkamzso revealed
numerous bacilli, micrococci, a small kind of yeast in state of
gemmation, and some mycelium of mucor. Tested with dilute
solution of methyl violet, many bacilli became coloured, and
were thus proved to be dead. ‘The iodine-test showed the pre-
sence of some still unchanged starch by the blue colouration of
many of the particles, and of undissolved proteids by the yellow
colouration of others.)

Upon chemical examination of the filtrate, I found, (besides
lactic acid), peptone, sugar, amido-acids, and slight traces of
ammonia, but I could not demonstrate the presence of enzymes.
Probably such were formed at first by the action of the bacteria,
but as the quantity of lactic acid increased, they must gradually
have been destroyed.“

Volatile acids were evidently present only in traces, and
certainly no butyric acid. Lactic acid was determined by titra-
tion with normal soda solution, and found to amount to 2.6
per cent.

The general composition of the sample analysed was as
follows :—

RNAI on, nih eae MME, Uadsce wan Sear 75 KON%G
Lactic acid Rem ay aes ace AROS
SUPA ue Mose pe a eees Gee - nn enw Bede %
odiumnChlonide sme i | aes hte | Oak %
Proteids

Amido-compounds |

Fats Ee Mise ti ics sas eee. LOANS,
Mineral matters |

Starch

My own observation on the fermentation of nukamiso has
convinced me that there is always simultaneous development of
bacilli and of a peculiar small kind of yeast, incapable of pro-
ducing alcoholic fermentation, and forming a scum, like that of

(1) An infection from a sample of nukamiso of sterilised one per cent peptone-
solution containing some extract of meat, showed after one week a great develop-
ment of bacteria, micrococci, mucor, and small yeast cells.

(2) The filtered liquid from nwkamiso was neutralised with carbonate of soda
and digested at 40° with meat-fibres as well as with boiled starch without any solu-
tion being observed.

218 INOUYE ; NOTE NO NUKAMISO.

Saccharomyces nrycoderma, on the surface, and that, after a few
weeks standing, not only sugar, carbon dioxide, and lactic acid are
formed, but also a considerable amount of proteids is dissolved.
If, at this time, a plate-culture is made with traces of the liquid,
bacterial colonies are observed, that liquefy gelatine, and also
colonies of the small yeast already mentioned."

The nwkamiso is prepared in barrels by almost every family
in this country, and is looked upon as indispensable for the
wants of the cuisine.

The products treated with it acquire a peculiar fine flavour.
The greater digestibility acquired is due to softening, and this
process is principally due to the death of the cells of the
vegetables caused by the lactic acid, which causes them to lose
their fullness. This process has here, therefore, the same effect
as the treatment with vinegar in the preparation, in western
style, of cucumber or lettuce-salads, or in the cooking of
vegetables ; but the experience in Japan is that the treatment
with nukamtso produces a superior result in regard to flavour.
It may be that nukamiso will gradually find its way into other
countries, as our soya-sauce or shoyu has done. '

(1) The colonies of a kind of Aspergillus, which I observed here, are probably
not a normal occurrence ; mucor was not observed in this case.

Preliminary Note on the Sake Yeast.
BY

K. Yabe, Nogakushi.

It has been repeatedly asserted that a fungus related to
Aspergillus called Eurotium Oryze, forms conidia, which can
bring on alcoholic fermentation. This fungus plays a great part
in the manufacture of sake on account of its high diastatic pro-
perties. Rice infected with that fungus, 7.c., koji, is mashed
along with freshly boiled rice, whereupon, not only sacchari-
fication, but also fermentation gradually sets in. Micros-
copical examination now reveals the presence of numerous cells
of a Saccharomyces, which have the size of the beer yeast,
but also frequently are about $ larger in diameter. We have
made several experiments in this Institute in order to see
whether this yeast could be transformed into the above named
mycelium fungus, and have always failed, from which it is to be
concluded that the sake yeast is not a certain stage of development
of Eurotium, but a species quite distinct. We hope to communi-
cate at a later occasion the details of further investigations.
For the present, the remarks which follow may suffice as to the
behaviour of the sake yeast.

This species exhibits its fermentative activity in solutions
containing a much higher percentage of alcohol than certain other
kinds of yeasts. A small amount of the yeast was suspended ina
Pasteur solution (containing a higher percentage of glucose than
usual) and quantities, of 10 cc. each, were placed in seven flasks,
to which were added 50 cc. of the Pasteur solution, a measured
amount of alcohol of known strength, and the necessary amount
of water to make the volume 100 cc. The percentage of sugar
in the flasks at the beginning was 9.48. After 6 days the sugar
was determined by Allihn’s method in those bottles where the
fermentation had been lively, while in others the usual titration
method was used. ‘The results are as follows :—

220 YABE ; NOTE ON THE YEAST.

Percentage of Sugar still Remarks.
alcohol. present,

) 0.024 Yeast multiplied very much.
4 0.098
8 1.649 >»

12 D-409 9

16 8.636 Multiplied but little.

20 9-254 Multiplied very little.

24 9.450 Any multiplication hardly observable.

We see, therefore, that the yeast was still very active in
presence of 12 % of alcohol, and even when there was 16 % some
fermentative activity was observed.

A large increase of mineral salts in the solution seems
also not to interfere, as is shown by the following results, obtained
with Pasteur solution to which increased quantities of sodium
chloride had been added.

Percentage of Percentage of sugar not Relative fermentative

sod. chlor. fermented after 2 weeks. power.
.0 0.0826 99.060
.005 0.0711 99.077
.050 0.0557 99-366

aut — —
5 0.0631 99.281
1.0 0.0655 99-255
2.0 0.0741 99-157
4-0 0.0753 99-134
6.0 0.1087 98.764
10.0 4.3982 48.868
12.0 7.5393 14.287
14.0 8.0580 8.391
18.0 8.6360 1.819

22.0 8.7660 fe)

Note on the Behaviour of Hippuric Acid in Soils.

BY

K. Yoshimura, Nogakushi.

The manurial effect of urine stands among other things in a
certain ratio to its amount of nitrogenous compounds, and to
the readiness with which these are converted into ammonia.

Kellner has shown that urea as such is not absorbed by
the soil and that surface soil more readily converts it into am-
monium carbonate than soil from a certain depth. Uric acid
can also readily undergo fermentation, with the production of
ammonia and carbon dioxide, and with simultaneous oxidation.)

C,H,N,0,+8H,0+ 30=4NH,+5CO.+4H,0.

But there exists a considerable amount of nitrogen in still
another form in the urine of cattle and horses, viz., as hippuric
acid.

About ro % of the total nitrogen of cattle-urine and about
2% of that of horse-urine are present in this form. As hippuric
acid evidently resists the fermentative action of microbes more
than urea or uric acid, it seemed to me of interest to observe the
behaviour of hippuric acid in soils.

At first, however, I tested soil in regard to its absorptive
power for this compound.

It seemed to me possible that soil rich in hydrated oxide of
iron might show some absorptive power, ferric hippurate being
insoluble. Two kinds of soils were tested; one, from the farm
of this College in Komaba, near Toky6, consists of volcanic
ashes and loam, and contains about 8 % of humus and 8-11 %
of oxide of iron; it has a high absorptive power for ammonia
and phosphoric acid. The jother, from Tochigi, is of clayey
nature; both soils are almost free from calcium carbonate.

(1) Sestini. Landw. Vers. Stat., 38., 157.

222 YOSHIMURA ; BEHAVIOUR OF

100 cc. of a solution of sodium hippurate, containing 0.340 %
hippuric acid, were well shaken with 20 grms. of soil from Komaba
and filtered off after 24 hours, without washing out the soil.
Of the filtrate 50 cc. were evaporated to a small volume and,
after addition of some sulphuric acid, shaken with acetic ether.
The latter after separation left on evaporating, 0.169 grm.=
0.338 % of hippuric acid. It may, therefore, be concluded that
practically none of it was absorbed.

Two experiments were then made with free hippuric acid
on both kinds of soils, but no absorptive power could be noticed
to any notable extent.

In regard to its behaviour towards fungi, I found that dilute
solutions of its sodium salt, containing neutral potassium phos-
phate and magnesium sulphate, are capable of developing mould
fungi and microbes, although this salt must be considered as a
poor nourishing material.

In order to see whether the microbes of the surface soil
would be more energetic in decomposing sodium hippurate than
those of the subsoil, I took portions of soil at different depths
reaching to 140 centimetres, shook them with sterilised water,
and infected a sterilised solution, containing 1 % sodium hippu-
rate, 0.2 % potassium phosphate and o.1 % magnesium sulphate.
The flasks, plugged with cotton, were allowed to stand from
December 29th, 1894, to May 5th, 1895.

From time to time, a few drops were withdrawn by means
of sterilised glass tubes and tested with Nessler’s reagent, and it
was thus found that, after two months, the solutions infected
with the soil from a depth of 50 centimetres, showed moderate
reaction, while those infected from greater depths, showed no
trace™ of it. But, later on, and when the temperature had
become warmer, increased vegetation of the microbes was noticed
in all the solutions, and by the 4th of May, 1895, a strong
ammoniacal reaction was obtained in all of them.

It was interesting to find nitrites absent; only in one flask
was a very slight trace indicated by Griess’s reaction.

(x) The temperature of the room was, until the end of February, generally lower
than 5 degrees C.

HIPPURIC ACID IN SOILS. 223

Several bottles were examined for undecomposed hippuric
acid, but none could be found.
The bacterial vegetation consisted mainly of micrococci.

Results.

Hippuric acid and its sodium salt are not absorbed by the
soils tested.

Decomposition of hippurates proceeds more quickly in the
surface soil than in the subsoil; this decomposition is attended
with liberation of ammonia, and is chiefly dependent upon the
action of micrococci.

Note to the Preceding.
BY

Prof. Oscar Loew.

The fact observed by Yoshimura that nitrification does not
take place in solutions of sodium hippurate is in accordance
with other similar observations. The nitrifying microbes, able
to assimilate ammonium carbonate, according to Huippe and
Winogradszki, cannot assimilate ammonium formate, and do
not develope well upon ammonium oxalate,’ as I have myself
ascertained. In my experiments, the sterilised solutions con-
tained, (besides 0.5 per mille of one of these salts), 0.5 per
mille each of neutral potassium phosphate, and magnesium
sulphate, and were infected from a culture obtained from garden
soil exhibiting a moderate nitrifying activity. The flasks, holding

(1) The objection that the want of nitrification is not under all conditions a
reliable sign of the absence of nitromonas, seems to me not tenable. In those cases,
however, in which other microbes are also present, the ammonium nitrite formed can
easily be destroyed.

224 LOEW ; BEHAVIOUR OF HIPPURIC ACID IN SOILS.

2 liters each, remained for 12 weeks, in the dark at 16-18°.
The fungoid mass which formed was exceedingly small and
hardly visible. While there was no nitrite formed in the flask
containing the formate,“ there was some in the oxalate flask.
A small portion of this liquid was diluted with g times its
volume of water, and the coloration obtained by Griess’s reaction
compared with that of standard, highly diluted nitrite solutions.
In this way, the conclusion was arrived at that the total
amount of nitrous acid did not exceed one milligramme, while
the control solution with ammonium carbonate contained more
than ro times as much. I observed further that the nitrification-
process proceeds nearly double as quickly in the dark as in

the daylight.

(1) There exists however a certain bacillus—I have called it Bacillus methylicus-
able to assimilate formates (Centrbl. f. Bact., #2, Nr. 14).

Does Hydrogen Peroxide occur in Plants?
BY

J. Cho.

It had been often stated that hydrogen peroxide in small
quantities occurs in plants, but it was shown by Th. Bokorny"? and
also by W. Pfeffer, that these statements are not well founded.
Quite recently, however, it has again been asserted to do so by
A. Bach.) After he had tried all known reagents and had found
them to be insufficient for proving the presence of very small
quantities of hydrogen peroxide in plants, he employed a new
reagent which, indeed, may prove useful in certain cases.

This consists of a highly diluted mixture of potassium bi-
chromate with free aniline.

The liquid to be tested has to be mixed with an equal
volume of this reagent, in presence of a little oxalic acid. Ifa
trace of hydrogen peroxide is present, a violet coloration is pro-
duced, due to the action upon aniline of perchromic acid,
intermediary formed.

I have convinced myself of the delicacy of this interesting
reaction. Bach employed it in examining twenty-five species of
plants, of which eighteen yielded a positive result. He says,

‘‘Sur les vingt-cing espéces végétales examinées, les
dix-huit suivantes ont donné un résultat positif en ce qui
concerne la présence de l’eau oxygénée :

Brassica asperifolia, B. oleifera, Daucus carota, Beta
vulgaris, Geranium rotundifolium, Hedera helix, Lauro-
cerasus, Aster, Tropceolum pentaphyllium, Chrysanthe-
mum JBalsamita, Mericurialis annua, Urtica, Calla
palustris, Vicia fava, Papaver rhoeas, Sisymbrium nastur-

(t) Pringsheims Jahrb., vol. 17.

(2) Ber. Sachs. Akad. Wiss., 1889. p. 493.

(3) Comptes rendus., t. CXIX., p. 286.

(4) The solution contains in one liter 0.03 gr. potassium bichromate and 5

drops of aniline. In testing, one drop of a 5 °/, solution of oxalic acid is added for
5 cc. of the reagent.

226 J; CHOY

tium, Dianthus Caryophyllus, Apium Petroselinum,
Fragaria vesca.

Deux plantes ont donné un résultat douteux: Lactuca
sativa, Vicia. Cinq plantes ont donné un résultat négatif:
Medicago sativa, Cichorium Intybus, Avena sativa, Viola
odorata, Lilium bulbiferum.”’

In regard to the mode of application he gives the follow-
ing note:

“*25 gr. de feuilles bien vertes et cueillies au milieu de la
journée ont été placés dans une tasse de porcelaine spa-
cieuse, arrosés par 75 cc. d’eau acidulée par 0.1 pour 100
d’acide oxalique.

Il faut éviter d’employer les acides mineraux, parce
qu ils décomposent certains glucosides et en mettent en
liberté la tannin.

La tasse, couverte @une soucoupe, a été gardée dans
une chambre obscure. A des intervalles déterminés, des
portions de 5 cc. ont été relevées sur l’extrait et essayées
par le réactif, comme il a été indiqué plus haut.

Un essai a blanc a été opéré dans chaque cas avec 5 cc.
de réactif, 5 cc. de l’eau acidulée et I ou plusieurs gouttes
de la solution d’acide oxalique.”

As it seemed to me of great interest to see whether the pre-
sence of hydrogen peroxide had thus been positively proved, I
subjected the following twenty-one species of plants exactly to
the same treatment as Bach:

Brassica oleracea. Geranium nepalense. Kerria japonica.

Brassica chinensis. Cornus officinalis. Rubus trifidus. +

Pisum sativum.-+ Corylopsis spicata. Prunus Lauro-cerasus.

Vicia fava. Urtica thunbergiana. Styrax obassia.

Papaver rhceas, Hydrangea hortensis.-+- Chrysanthemum coronarium.+
Diervilla grandiflora. Daucus carota.+- Aster tartaricum.+

Lonicera morrowii. Cryptotenia canadensis. Beta vulgaris.+-

In nine cases out of these twenty-one, I did, indeed, observe
a reddish coloration, but it was not like that in the control
test. I have marked these species in the list with+.

I found that in these cases parts of the leaves had died
from the poisonous action of the oxalic acid, while in the other
cases, where no reaction was obtained, all the leaves were still

DOES HYDROGEN PEROXIDE OCCUR IN PLANTS, 227

uninjured, and the cuticula had prevented the entrance of
the acid solution into the cells. In order to see whether the
reaction obtained was, indeed, due to hydrogen peroxide, I
added 0.5 gr. of platinum black to 20 cc. of the diluted oxalic
acid solution that had been in contact with the leaves for twenty-
four hours, and let the mixture stand with occasional stirring in
a porcelain dish for one hour. A control experiment with a
o.1 % solution of hydrogen peroxide, having shown that by this
means hydvogen peroxide) is so perfectly decomposed, that
Bach’s reaction shows not a trace of it, it was remarkable to find
that in all the plants mentioned the reddish coloration was obtained,
and still of the same intensity as before treatment with platinum black.
The only conclusion to be drawn is that there was no hydrogen
peroxide ever present, and it seems to me that Bach was not
justified in declaring that it was so in any of the plants he ex-
amined, because he omitted to apply the platinum test to see
whether the faculty of giving this reaction would be destroyed
or not.

The coloration mentioned can very probably be obtained
only in those cases where the leaves have been partially killed
by the oxalic acid solution, and thus certain easily oxidisable
organic matters made able to leave the cells by osmosis. ‘These
might by oxidation in presence of the aniline oxalate yield
coloured products.

I doubt whether Bach ever observed his reaction in leaves
that had remained alive in all parts.

(1) The energetic decomposition of hydrogen peroxide by platinum black is an
old and well known fact.

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Die Japanischen Laubholzer im Winterzustande.

Bestimmungstabellen
VON

H. Shirasawa, Ringakushi.

VORWORT.

SCHWARZ hat in seinem ‘‘ Forstliche Botanik”’ die Unter-
scheidungsmerkmale angegeben, wonach die forstlich wichtigen
Pflanzen in Deutschland auch ohne Bliithen oder Friichte oder
im ganz kahlen Zustand zu erkennen sind.

Eine Bearbeitung der japanischen Pflanzen in dieser Hin-
sicht liegt noch nicht vor, was als eine oft schwer gefiihlte
Liicke in unserem forstlichen Wissen zu betrachten ist, und
mich darum veranlasst hat, die vorliegende Arbeit durchzufiih-
ren. Ich bin dabei hauptsachlich dem System von Schwarz
gefolgt, so weit dies bei der naturgemdss grossen Verschieden-
heit der deutschen und japanischen Flora moéglich war.

Die zu dieser Arbeit verwendeten Examplare (von ungefahr
300 Arten, Baum- Strauch- und Schlinggewachsen) wurden
meist in den botanischen Gdrten in Komaba und Koishikawa,
und dann in der Umgebung von Tokyo gesammelt. Da die im
Garten gezogenen Pflanzen meist kein normales Wachsthum
zeigen, so habe ich nebenbei auch frei im Walde erwachsene
Pflanzen in Amagi, Nikko, und in der Provinz Shinano zur
vergleichenden Betrachtung zugezogen.

Zu dieser Arbeit wurden zwei Winter benutzt. Bei dieser
Kiirze der Zeit konnten im Nachfolgenden noch nicht alle forst-
lich wichtigsten Pflanzen Japans bearbeitet werden. Indessen
behalte ich mir vor, im nachsten Winter diejenigen Pflanzen,
welche noch nicht beriicksichtigt worden sind, naher zu
untersuchen.

Ich kann hier nicht umhin den meinen warmsten Dank
Herren Dr. Grasmann, Prof. Matsumura, Assist. Prof. Shirai u.

| fare

230 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Dr Honda, Prof. Dr. Kitao u. Prof. Dr. Ishikawa auszusprechen

welche mir ihre giitige Hiilfe bei der vorliegenden Arbeit hatten
zu Theil werden lassen.

Tokyo Juni, 27 Meiji.

TABELLE I.

Die Knospen sind an den Langtrieben spiralig
angeordenet.

I.—Die Knospen sind in der Blattnarbe verborgen.
Robinia Pseudacacia, L. WHari-enju. Taf. I, Fig. r.

Die Blattnarbe ist ziemlich gross, hockerig, wird zumiest
von zwei derben geraden Stacheln flankiert. Die Stacheln
konnen auch fehlen. Zweige sparrig, hin- u. hergebogen,
kantig, lang. Rinde braun od. griinlichbraun mit zahlreichen,
feinen Lenticellen, im Alter braungrau bis grau. Mark un-
regelmassig eckig, ziemlich weit (nach Schwarz).

II.—Knospen gestielt.

a.—Eine gréssere Schuppe umhiillt fast die ganze Knospe.

Alnus incana, Willd. var. glauca, Ait. Yama-hannoki.
Taf. I, Rigs:
Knospen ziemlich gross eiformig, schwach gekriimmt, un-
deutlich dreieckig, lang gestielt. Knospenschuppe dunkel-
carminroth, bereift. Zweige hin- u. hergebogen, harzig. Die
jtingeren Theile der Triebe sind feinfilzig behaart, grau, u.
lassen die weissflichen Lenticellen nicht so deutlich erkennen.

Mark dreieckig.
Alnus japonica, Szeb. ef Zucc. WHannoki. Taf. I, Fig. 3.

Knospen schlank, lang, unregelmdssig dreieckig, scharf
kantig, etwas gefaltet. Die Spitze der Knospen schwach ge-

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 231

kriimmt, harzig, glanzend. Jungtrieb dreikantig graubraun
bereift, altere grau. Lenticellen ziemlich erkennbar. Mark
dreieckig.

Alnus firma, Sieb. et Zucc. Yashabushi. Taf. I, Fig. 4.

Knospen spindelformig, gekriimmt, kurz gestielt, rothbraun
glanzend, theilweise griinlichgelb. Blattnarbe abgerundetes
Dreieck. Zweige hin- u. hergebogen, auf der Sonnenseite
braun, 2uf der Schattenseite graubraun, die jiingeren Theile
fein behaart. Lenticellen sehr deutlich. Mark unregelmas-
siges Dreieck.

Alnus viridis, DC. var. sibirica, Regel. Miyama-hannoki.

Taf. I, Pig. 5.

Knospen ziemlich gross, spindelig, lang gestielt, zugespitzt

u. gekriimmt. Knospenschuppen dunkelcarminroth, glanzend.

Die jungen Zweige graubraun, kantig, mit elliptischen, deut-

lichen Lenticellen versehen, die alteren Zweige dunkelgriin,

mit braunweisslichem Ueberzug. Blattenarbe halbkreis- od.
herzformig, u. von unregelmassiger Form.

b.—Mehrere Schuppen umgeben die Knospen spiralig.

Ribes} fasciculatum, Sieb. et Zucc. Yabu-sanzashi. Taf. I, Fig. 6.
Knospen spindelformig, lang, etwas gebogen, den Zweigen
angedriickt, hellroth gefarbt. Jiinge Triebe hellbraun, glan-
zend, bei Absterben der ausseren Peridermschichten grau, u.
leicht sich abschifernd. Blattnarbe sichelformig. Lenticellen
undeutlich. Mark ziemlich weit.

III.—Knospen von unausgebildeten Blattchen gebildet,
(nackt),

A.—Zweige auffallend dick.
a.—Mark gefachert.

Juglans Sieboldiana, Max. Oni-gurumi. Taf. I, Fig. 7.
Endknospe viel grésser als Seitenknospe, pyramidal, etwas
gebogen, graubraun filzig behaart. Junge Zweige braun be-
haart, altere weisslichgrau, kahl. Blattnarbe sehr gross breit

232 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

herzformig. Mark weit fiinfeckig, gefachert. Bei Verwundung
der Basttheile zeigt sich gelbe Farbung.

Juglans cordiformis, Max. Hime-gurumi. Taf. I, Fig. 8.

Knospen u. Zweige ganz ahnlich wie bei der vorhergehenden
Art, daher die Unterscheidung im Winterzustande schwer.

Pterocarya rhoifolia, Sveb. ef Zucc. Sawa-gurumi.
Taf. Ty Piagwizo:
Knospen dreikantig, lang, auf dem Langschnitte spatelfér-
mig, an der Spitze etwas gedreht, graubraun filzig behaart.
Zweige gerade, ganz kahl, die jiingeren Theile hellbraun od.
grau. Mark ziemlich weit, eckig, gefachert.

Juglans regia. L. var, sinensis, Cas. ‘Teuchi-gurumi.
Taf. I, ite:
Endknospen grosser als Seitenknospen, glockenformig, etwas
kantig, dunkelgrau filzig behaart; die ausseren 2 od. 4 elgen-
tlichen Schuppen leicht ablésbar. Seitenknospen kugelig.
Zweige dick, die jiingeren graulich braun od. griin; die alteren
grau glanzend. Wenige Lenticellen, an jiingeren Zweigen
deutlicher als an alteren. Blattnarbe gross, breitherzformig.
Mark gross etwas gefarbt.

£.—Mark nicht gefachert.

Cedrela chinensis, fuss. Chanchin. Taf. II, Fig. zo.

Endknospe sehr viel grdsser als Seitenknospen, von
mehreren unausgebildeten Blattchen locker umhiillt, dunkel-
braunfilzig behaart. Seitenknospen kugelig. Zweige dick,
dunkelgelbbraun, glaénzend, schwarz bestaéubt. Blattnarbe
gross, rundlich, grauweiss, mit meist 5 Gefassbundelspuren
versehen. Lenticellen wenig, deutlich. Mark weit, rundlich.

Rhus vernicifera, DC. Urushi. Taf. I, Fig. 72.

Endknospen viel grésser als Seitenknospe, pyramidal, an der
Spitze etwas gekriimmt. Zwei od. drei grosse unausgebildete
Blattchen umgeben fast die ganze graufilzig behaarte Knospe,
Seitenknospen kugelig. Zweige dick, grau, glanzend, mit
mehreren Lent. versehen. Blattnarbe breit herzformig QQ.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 233

Mark weit. Beim Abschneiden der Rinde fliesst gummiartige
Fliissichkeit aus, giftig.

Rhus trichocarpa, Mig. Yama-urushi. Taf. I, Fig. 74.

Endknospe grosser als Seitenknospe, pyramidal, braunfilzig
behaart. Seitenknospen etwas flach. Zweige weniger dick
als die Rhus virnicifera, mit vielen, feinen Lenticellen, dun-
kelgrau gefarbt. Blattnarbe gross, ein gleichschenkliges Drei-
eck bildend, glanzend, réothlich gefarbt. Mark weit.

Rhus succedanea, L. Haze. Taf. I, Fig. rz.

Endknospen sehr gross, pyramidenformig, von unausgebilde-
ten Blattchen auf drei Seiten dicht umbiillt; sie sind hell-
braufilzig behaart. Seitenknospen klein, keilformig. Zweige
graubraun. Blattnarbe halbmondformig. Mark weit, etwas
eckig.

Rhus sylvestris, Szeb. et Zucc. Yama-haze. Taf. I, Fig. 73.

Knospen ahnlich wie bei der vorhergehenden Art, aber
locker umhiillt, von unregelmassiger Form, gelblichbraun filzig
behaart. Seitenknospen flach, u. lang. Zweige dunkelgrau,
schmutzig gefarbt. Blattnarbe wie bei Rhus vernicifera.
Mark weit.

B.—Zweige weniger dick.
a.—Zweige ohne Stacheln od. Dornen.

Halesia hispida, Benth. et Hook. Ke-asagara. Taf. I, Fig. 16.

Knospen spindelig, gelblichgrau od. gelb gefarbt. Endknos-
pen grosser als Seitenknospen, etwas gebogen, lang. Blattnarbe
gross schildformig. Seitenknospen parallel stehend an den
Trieben. Junge Zweige hellgelbbraun, altere braun. Lenti-
cellen deutlich. Mark ziemlich weit, eckig. Die Rinde hat
eigentiimlichen Geruch.

Halesia corymbosa, Benth. et Hook. Asagara. Taf. I, Fig. 77.

Zwei unausgebildete Blattchen umgeben die eifdrmige,
dunkelbraune, kleinwalzige Knospe. Zweige hin u. hergebogen,
braun, kleinwalzig, dltere dunkelgraubraun. Blattnarbe gross
vier eckig. Lent. undeutlich. Mark rund.

234 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

Ramnus crenata, Sieb. ef Zucc. Isonoki. Taf. I, Fig. ro.

Endknospen groésser als Seitenknospen, pyramidenformig, an
der Spitze gekriimmt, od. abgeflacht, graubraunfilzig behaart.
Zweige schlank, jiingere Theile behaart, dunkelbraun. Lenti-
cellen deutlich. Mark rund.

Meliosma myriantha, Sieb. et Zucc. Awabuki. Taf. I, Fig. 20.

4 od. 5 pfriemenformige, unausgebildete Blattchen an der
Spitze der Triebe zusammensitzend, braunfilzig behaart. Blatt-
narbe halbelliptisch, u. die Gefassbiindelspuren U-formig ange-
ordnet. Zweige dunkelrothbraun, auf der Schattenseite grau.
Lent. deutlich. Mark elliptisch.

Mallotus japonica, Muell. Arg. Akame-gashiwa. Taf. I; Fig. 78.

Knospen von 4 od. 6 unausgebildeten, gelblichbraunen, weiss-
lich bereiften Blattchen umbhiillt, wovon zwei gegenstandig
sind meist grésser als die anderen. Endknospen wesentlich
grosser als Seitenknospen, die letzteren parallel stehend an
der Triebe meist mit Nebenknospen. Zweige dick, dunkel-
braun, die jiingeren Theile bereift, kantig. Lenticellen un-
deutlich. Mark sehr weit.

b.—Zweige mit Stacheln od. Dornen.
a.—Zweige mit Stacheln.
Caesalpinia sepiaria, Rox). Jaketsu-ibara. Taf. II, Fig. 3, 4.

Knospen sind von zwei unausgebildeten Blattchen umgeben,
u. tiber den Blattachseln etwas entfernt stehend. Zweige
ziemlich dick, mit zahlreichen, derben zuriickgebogenen
Stacheln auf der ganzen Flache der Triebe, gelblichbraun od,
braun gefarbt. Blattnarbe gross. Lent. deutlich. Mark weit.
etwas gefarbt.

£.—Zweige mit Dornen.
Elaeagnus umbellata, Thunb. Aki-gumi. Taf. II, Fig. 2.

Endknospen etwas grésser als Seitenknospen, pyramiden-
formig. Seitenknospen etwas flach, an den Trieben senkrecht
stehend. Zweige u. Knospen mit kleinen, zusammengedrtick-
ten grauweissen Schuppen bedeckt. Zweige schlank. Blattnarbe
halbmondfoérmig. Lent. undeutlich. Mark weit, rund.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 235

Elaeagnus longipes, A. Gray. Natsu-gumi. Taf. IT, Fig. z.
Endknospen dreikantig, nicht zugespitzt. Seitenknospen
etwas flach. Zweige u. Knospen sind mit kleinen zusam-
mengedriickten, braunen, od. dunkelbraunen Schuppen bedeckt,
an der ersteren sind starkere Dornen vorhanden. Blattpolster
hoch vorspringend; Blattnarbe rundlich, od. halbmondformig.
Mark sehr weit, rundlich.

IV.—Knospen sitzend, so klein, dass man die Stellung
u. Zahl der Schuppen nicht mehr deutlich wahr-
nehmen kann.
a.—Zweige mit metamorphosierten Nebenzweigen.
Lycium chinense, Mill. Kuko.
Knospen kugelig. Schuppen braun. Zweige diinn, grau, bei
zunehmender Dicke mit 5 zeiligen Korkleisten versehen. Mark
weit. Rinde hat eigentiimlichen unangenehmen Geruch.

f.—Zweige mit Dornen.
Zizyphus vulgaris, Lam. Natsume. Taf. II, Fig. 5.

Knospen ziemlich gross, meist unter der halbmondfomigen
Blattnarbe sitzend. Junge Zweige lang gestreckt, dunkelgelb-
lich braun, altere Zweige schwdarzlich grau mit grossen
Kurztrieben. Zweige und Kurztriebe deutlich von einander
verschieden. Stacheln schlank, je zwei neben jeder Knospe.
Lent. fein, wenig deutlich. Mark von unregelmadssiger Form.
Holz gelblich.

Paliurus aubletia, Roem et Sch. Hama-natsume. Taf. II, Fig, 6.
Knospen klein, gelblich braun, behaart. Zweige hin- u.
hergebogen, gelblich- od. graéulichgrtin, bereift. Stacheln lang

u. derb, glanzend dunkelbraun, je zwei neben den Knospen
stehend. Lenticellen deutlich. Mark rundlich.

7-—Zweige ohne metamorphosierten Nebenzweigen oder
Dornen.
Ilex Sieboldii, Mig. Umemodoki. Taf. II, Fig. &.
Knospen kegelig, etwas zugespitzt. EEndknospe grosser als
Seitenknospen, jede mit einer Nebenknospe auf der unteren

236 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

Seite. Knospenschuppen schwach behaart. Zweige hin- u.
hergebogen, dunkelgrau. Blattnarbe halbmondférmig. Lenti-
cellen zahlreich, deutlich. Mark schmal, unregelmassiger
Form.
‘Tlex geniculata, Max. Furin-umemodoki. Taf. II, Fig. 7.
Knospen wie bei der vorhergehenden Art. Zweige dunkel-
carminbraun, glanzend, gerade. Blattnarbe halbkreisformig.
Die jiingeren Zweige unterscheiden sich von den 4lteren durch
ganz verschiedene Farbung. Mark zeimlich weit, von unregel-
mdassiger Form.
Hibiscus syriacus, L. Mukuge.
Knospen grau behaart. Zweige grau, od. dunkelgraubraun,
mit feinen Lenticellen, bereift. Blattpolster etwas vorsprin-
gend. Blattnarbe rundlich od. halbmondformig. Mark rund.

V.—Knospen sitzend, Zahl der Schuppen od. kleinen
zusammengedruckten Blattchen undeutlich.
A.—Einjahrige Zweige auffallend dick.
a.—Zweige ohne Stacheln od. Dornen.

a.—Knospen filzig behaart.

Ailanthus glandulosa, Desf. Shinju.

Knospen gleichgross, verhaltnissmadssig klein, halbkugelig,
roth, weisslich behaart. Blattnarbe sehr gross. Zweige un-
regelmassig gebogen. Die einjaéhrige Triebe sehr fein od.
dicht behaart, hellbraun, die altere grau bis braun. Mark
sehr weit, rund.

Melia japonica, G. Don. Sendan. Taf. II, Fig. 9.

Knospen kugelig, hellbraun dicht behaart. Einjahrige
Zweige dick, dunkelgriin, mit vielen deutlichen Lenticellen,
bereift. Blattnarbe etwas vorspringend. Mark weit, rundlich.
Holz gelb gefarbt.

Rhus semi-alata, Murr. var. Osbeckii. DC. Nurude.
Taf. TI, Fag 2:
End- u. Seitenknospen gleich gross, mit weissbrauner,
baumwolle-artiger, dichter Behaarung. Zweige hellgraubraun,
schwach glanzend, gerade, schwarzlich gefleckt. Blattnarbe

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 237

gross, umfasst fast die ganze Knospe, von nebenstehender
Form, Y. Lenticellen deutlich. Mark sehr weit, etwas
gefarbt.

£.—Knospen nicht od. wenig behaart.

Ehretia acuminata, R. Br. Chishanoki. Taf. II, Fig. z3.
Knospen auf dem Zweige ganz dicht sitzend, von kurzer
stumpfer Kegelform. Zwei graue, od. dunkelbraune wenig
behaarte Schuppen umhiillen die Knospe von beiden Seiten.
Zweige ziemlich dick, grau, glanzend, schwach behaart, fiihlen
sich rauh an. Lenticellen wenig, deutlich. Blattnarbe rund-
lich, od. halbelliptisch. Mark sehr weit, rundlich.

Sapindus Mukurosi, Gaertn. Mukuroji. Taf. I], Fig. rz.
Knospen dicht sitzend, halbkugelig, 4 Schuppen umgeben
ganz dicht die Knospe. Blattnarbe gross herzférmig. Zweige
dick, graugriin, schmuzig bestaubt. Lenticellen zahlreich,
deutlich. Mark weit, gefarbt. Rinde gelblich.

b.—Zweige mit Stacheln oder Dornen.

Zanthoxylum ailanthoides, Sich. et Zucc. Karasuno-sansho.
Taf, III; Fig.3.
Knospen beulenformig, ganz dicht an der Triebe sitzend, so
dass man die Zahl der Schuppen nicht deutlich wahrnehmen
kann. Zweige affallend dick, dunkelgraubraun mit zahlreichen
Stacheln. Blattnarbe gross, rundlich. Mark weit, gefachert.
Die Rinde hat einen eigenthtimlichen Geruch.

B.—Einjahrige Zweige weniger dick.
a.—Zweige ohne Stacheln od. Dornen.

a.—Knospen nicht od. wenig behaart.

Albizzia Julibrissin, Boiv. Nemu. Taf. II, Fig. 26.
Knospen klein kegelformig, etwas flach, iiber der dreieckigen
Blattnarbe sitzend. Zweige hin- u. hergebogen; die jiingeren
_graubraun schwarz bestadubt, schmutzig. Lenticellen deutlich.
Mark weit, strahlenformig.

238 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

8.—Knospen behaart.
Sophora japonica, L. Enju. Taf. II, Fig. 77.

Knospe klein, von dem Blattpolster umfasst, u. im inneren
Theile derselben sitzend. Schuppen undeutlich, mit dunkel-
carminvioletter Behaarung. Ejinjahrige Zweige griin, auf
der Sonnenseite gelb od. braunlichgriin, die alteren Zweige
grau. Die ausseren Peridermschichten reissen in Langsstrei-
fen auf. Markrundlich. Die Rinde hat einen eigenthiimlichen
Geruch.

Marlea platanifolia, Sieb. et Zucc. Urinoki. Taf. II, Fig. 20.

Knospen entwickeln sich aus der Mitte der Blattnarbe,
graubraunfilzig dicht behaart, kugelig mit kleinen Nebenknos-
pen auf der unteren Seite. Zweige braun, gerade, hin- u,
hergebogen. Lenticellen zahlreich. Mark weit, etwas
gefarbt. Rinde mit Geruch. .

Styrax Obassia, Szeb. et Zucc. Hakuun-boku. Taf. I, Fig. 22.

Knospen entwickeln sich aus der Mitte der Blattnarbe,
eiformig, lang, etwas flach, mit kleinen Nebenknospen, gelb-
braunfilzig behaart. Zweige gerade, hin- u. hergebogen, die
jiingeren rothbraun glanzend, glatt, die alteren dunkelbraun.
Blattnarbe elliptisch. Lenticellen undeutlich. Mark rund.

Styrax japonica, Sieb. et Zucc. Egonoki. Taf. II, Fig. ar.

Knospen weniger gross, eiformig etwas kantig, an der Spitze
gekriimmt, dichtfilzig behaart. Zweige diinn besenformig, die
jiingeren hellbraun glanzend, bei der Alteren die dusseren
Peridermschichten in Langenstreifen aufreissend, so dass die-
selben mit faserigen Peridermschichten bekleidet erscheinen.
Lenticellen undeutlich. Mark klein, rund.

b.—Zweige mit Stacheln od. Dornen.
a.—Zweige mit Dornen.
Cudrania triloba, Hance, Hari-guwa. Taf. II, Fig. 78, 79.

Knospen dicht sitzend an der Triebe, beulenformig, Schuppen
undeutlich, dunkelgriinbraun gefarbt. Die einjahrigen Zweige
griin od. graugriin mit senkrecht stehenden starken Dornen.
Blattnarbe halbmondformig. Lenticellen ganz deutlich. Mark
rund.

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 239

Gleditschia japonica, Mig. Saikachi. Taf. II, Fig. 74, 75.
Die Halfte der Knospen von der Rinde umgefasst, so dass
man die Knospenschuppen nicht ganz deutlich wahrnehmen
kann. Zweige griin, glanzend, ziemlich dick, mit starken, ver-
zweigten Dornen. Die einjahrigen Zweige mit leicht abschilf-
baren, feinen Peridermschichten. Blattnarbe von der Form,
Lenticellen sehr deutlich, rund. Mark eng.

&.—Zweige mit Stacheln.
Zanthoxylum piperitum, DC. Sansho. Taf. II, Fig. 23.
Knospen von zusammengedriickten Blattchen bekleidet ; die
Seitenknospen kugelig. Die Zahl der Blattchen undeutlich,
braun. Zweige schwarzbraun. Auf jeder Seite der Knospe
. Zwei Stacheln. Blattnarbe halbmondformig. Lenticellen
deutlich. Mark rund. Die Rinde hat einen eigenthiimlichen
Geruch, u. scharfen Geschmack.

Zanthoxylum alatum, Roxb. Fuyu-zansho. Taf. III, Fig. 2.

Endknospe lang gestreckt, Seitenknospen kugelig, Schuppen-
blattchen wenig deutlich, dunkelgriin gefarbt. Stacheln sind
sehr breit u. lang, rothbraun glanzend. Zweige wie Stachein
gefarbt. Blattnarbe ziemlich gross, halbmondformig od. halb-
elliptisch. Mark weit, rund. Die Rinde hat einen eigen-
thiimlichen Geruch, aber weniger scharf als Zan. piperitum.
Im Winter verbleiben einige griine Blatter.

Zanthoxylum schinnifolium, Szeb. et Zucc. Inu-zansho.
Naje LG Bie a.
Seitenknospen gegen Triebe zu abgeplattet. Knospenschup-
pen undeutlich. Zweige dunkelbraun, schwarz bestaubt. Nur
einzelne Stacheln langs des Triebes vorhanden. Blattnarbe
halbelliptisch. Lenticellen deutlich. Mark weit, rund.

VI.—Knospen sitzend mit einer bezw. zwei Schuppen.
A.—Knospen ausgesprochen kegelformig, von der Blatt-
narbe ringformig umgeben.
a.—Knospen od. junge Zweige behaart.

Salix brachystachys, Benth. Osaruko-yanagi. Taf. III, Fig. ee
Blitenknospen gross spindelformig; Laubknospen kleiner,

240 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

kegelig, gegen die Tricbe zu geplattet. Nur eine roéthlich
gefarbte, u. weiss od. grau behaarte Schuppe umhiillt die
ganze Knospe. Zweige rothlich, dunkelgrau behaart. Lenti-
cellen wenig. Mark weit, eckig.

b.—Knospen od. Zweige nicht behaart.
Salix gracilistyla, Mig. Kuro-yanagi. Taf. III, Fig. 6.

Knospen kegelformig, gegen die Zweige zu abgeplattet.
Bliitenknospen spindelig, lang, an der Spitze etwas gebo-
gen. Zweige u. Knospenschuppen dunkelrothlich, glanzend.
Man kann diese von anderen Arten leicht unterscheiden,
durch die rothlichen, auf der Lichtseite aber schwarzlichen
Bliitenkatzchen.

Salix japonica, Thunb. Shiba-yanagi. Taf. III, Fig. 8, 9.

Knospen kegelformig, lang, an der dem Treibe zugewendeten
Seite abgeplattet. Bliitenknospen grosser als Blattknospen, an
der Spitze gekriimmt. Einjahrige Zweige u. Knospenschuppen,
auf der Lichseite réthlich gefarbt, glanzend, altere Zweige
graubraun, hin- u. hergebogen. Blattpolster vorspringend.
Lenticellen wenig, deutlich. Strauchartig, nicht aufrecht
stehend. ;

Fieus ecarica, L. Ichijiku. Taf. III, Fig. 4.

Endknospe kegelig, an der Spitze etwas gebogen. Eine
grosse, griinlichgelbe Schuppe umnhiillt fast die ganze Knospe.
Seitenknospen halbkugelig, zugespitzt, von mehreren Schuppen
umgeben, schwarzlichbraun. Zweige dick, hin- u. hergebo-
gen, dunkelbraun. Blattnarbe rundlich, Mark etwas eckig
(sechseckig). Die verholzten ‘Triebe von unreglmassiger
Form. Bei Verwundung der Basttheile fliesst milchartiger
Saft aus.

B.—Knospen nicht kegelformig, stehen tiber der Blatt-
narbe. Nur eine beiderseits kantige Schuppe (aus
zwei Schuppen verwachsen).

a.—Zweige dottergelb.
Salix purpurea, L. Kawa-yanagi. Taf. III, Fig. zz.
Knospen sind an der Spitze der Zweige kleiner als in der
Mitte derselben. Die letzteren sind circa 2-3 mal so lang

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 241

als breit, die rothlichbraun behaarten Knospen stehen an dem
Zweige sehr dicht. Blattpolster etwas vorspringend. Seiten-
zweige sehr diinn, gelblich bis griinlichgrau, behaart.

Salix sp. (?)

Knospen halbkugelig od. etwas lang gestreckt. Bltitenknos-
pen wesentlich grosser, eiformig, an der Spitze gekriimmt,
gekielt. Zweige u. Knospenschuppen dottergelb, glanzend.
Blattpolster vorspringend. Blattnarbe von der Form @&.
Mark ziemlich weit, etwas eckig.

b.—Einjahrige Zweige roth od. braun, glanzend wie
lackiert.

a.—Knospen dichtfilzig behaart.

Hovenia duleis, Thunb. Kenponashi. Taf. III, Fig. 78, 79.
Zwei Knospen aufeinander sitzend, die untere Nebenknospe
kleiner als die obere, die letztere kegelférmig, etwas kantig,
schwarlich braun behaart. Die einjahrigen Zweige stark
hin- u. hergebogen, auf der Lichtseite dunkelbraun. Lenti-
cellen ziemlich deutlich. Mark eng, rund.

Magnolia Kobus, DC. Kobushi. Taf. III, Fig. x5.

Knospen eiformig, lang gestreckt, an der Spitze etwas
gekriimmt (Endknospen grosser als Seitenknospen.) dunkel-
grau, schwarz behaart. Zweige dunkelrothbraun, auf der
Schattenseite gelblich griin. Blattnarbe sichelformig, schmal.
Lenticellen deutlich. Mark gross u. rund. Die Rinde hat
eigenthiimlichen, angenehmen Geruch.

Magnolia obovata, Tiunb. Mokuren. Taf. III, Fig. rg.

Knospen eiférmig ziemlich lang, beiderseits abgeplattet,
etwas kantig u. gekriimmt. Knospenschuppen grauweissfilzig
behaart. Bliitenknospen bedeutend grosser als Laubknospen,
an der Spitze der Triebe sitzend. Blattnarbe von der Form
eines abgerundeten Dreiecks. Zweige schwarzlich braun.
Mark gross, rund. Lenticellen. deutlich. Die Rinde hat
eigenthiimlichen Geruch, jedoch schwacher als die vorige Art.

242 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE,

#.—Knospen nicht behaart,
Andromeda ovalifolia, Wall. Kashioshimi. Taf. III, Fig. 26, r7.

Knospen eiformig, etwas flach. Schuppen roth, glanzend.
Einjahrige Zweige roth gefarbt, glanzend wie lackiert ; altere
dunkelgrau. Blattnarbe halbmondformig, in der Mitte mit
einer einzigen Gefassbiindelspur. Lenticellen undeutlich.

Mark elliptisch.

Stachyurus praecox, Szeb. et Zucc. Kifuji. Taf. III, Fig. 2z.
Endknospen kugelig zugespitzt. Seitenknospen eiférmig,
gespitzt, an der Triebe angedriickt. Schuppen dunkelroth-
braun. Zweige von derselben Farbe, glanzend. Lenticellen
sehr deutlich. Mark rund, u. sehr weit.

Salix Thunbergiana, Neko-yanagi. Taf. III, Fig. 72.

Knospen halbkugelig. Nur eine, dunkelrothglanzende
Schuppe umgibt fast drei ganze Knospe. Bliitenknospen
eiformig, gross, gekielt. Zweige auf der Lichtseite dunkelroth,
auf der Schattenseite gelbgriin. Blattpolster vorspringend.
Blattnarbe schmal, umfasst die Halfte der Knospe. Lenticellen
sehr wenig. Mark ziemlich weit.

Itea japonica, Oliv. Zuina. Taf. III, Fig. 20.
Hat eigentlich drei Schuppen, wird daher in VII, A, b.
beschreiben.
Helwingia ruscifolia, Willd. Hanaikada. Taf. V, Fig. 7, 8.

Endknospen pyramidenformig, grésser als Seitenknospen.
Die letzteren flach, keilformig. Schuppen rothlich griin, kahl.
Zweige haben dieselbe Farbung od. gelblichgriin, glanzend.
Blattnarbe fast halbkreisformig. Lenticellen deutlich,
vereinzelt. Mark weit, rund.

c.—Zweige nicht auffallig gefarbt, Knospen lang gestreckt.
a.—Knospen nicht behaart, walzig.
Magnolia hypoleuea, Sieb. et Zucc. Honoki. Taf. III, Fig. 22.
Knospen sehr gross, spindelig, von gleichem Durchmesser
wie die Triebe, 4 cm. od. dariiber lang. Die zwei Knospen-
schuppen sind verwachsen, u. zeigen an der Verwachsungsstelle
eine Naht. Zweige dick, lang gestreckt, aufrecht stehend,

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 243

ohne Nebenzweige. Die einjaéhrigen Zweige griinbraun. Lent.
deutlich, lang. Mark rund. Die Rinde hat einen eigen-
thiimlichen, angenehmen Geruch.

8.—Knospen sehr wenig od. gar nicht behaart,
zugespitzt.
Lindera hypoleuca, Max. Kuromoji. Taf. VI, Fig. 4, 5.

Knospen spindelig, lang; Schuppen réthlichgelb wenig
behaart. Zweige glatt, auf der Lichtseite rothlich, auf der
Schattenseite gelbgriin, die alteren schwarzlich. Blattnarbe
von der Form, WV. Lenticellen sehr wenig. Mark rund.

Die Rinde hat einen eigenthtimlichen, angenehmen Geruch.

y-—Knospen behaart, dem Stengel angedriickt.
Salix viminalis, 2. Kinu-yanagi. Taf. III, Fig. 73.

Knospen sehr verschieden gross, dem Zweige dicht ange-
driickt, die Spitze etwas gebogen, (besonders bei den grossen
Knospen). Die jiingsten Zweigteile dichtfilzig behaart, altere
rothlichgelb, glanzend. Zweige oft von grosser Lange.

d.—Zweige griin od. braun; Knospen (vergekehrt) keil-
formig.
a.—Blattnarbe rundlich.
Diospyros Kaki, L. fil. Kaki. Taf. Ill, Fig. 23.

Knospen keilformig; Schuppen (oft zwei od. mehr) _hell-
braun, schwach behaart. Zweige ziemlich dick, hellbraun;
die jiingeren schwach behaart. Blattnarbe gross, rundlich,
gewolbt. Lent. deutlich. Mark von unregelmdssiger Form.

Diospyros Lotus, L. Mame-gaki. Taf. III, Fig. 24.

Knospen keilformig, lang; die Spitze etwas gebogen. Schup-
pen griinbraun, schwarzlich gefleckt, wenig behaart. Zweige
griinbraun etwas glanzend, schmutzig.

Broussonetia Kasinoki, Sieb. Kozo. Taf. IV, Fig. 2.

Knospen keilformig, kurz, aufrecht stehend obenhalb der
Blattpolster, Knospenschuppen dunkel od. schwarzbraun,
schwach behaart. Junge Zweige dunkelbraun od. schwarz-
braun, schmutzig. Lent. vorspringend, sehr deutlich. Mark
rund, weit.

244 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Broussonetia Kasinoki, Sich. vay. Hime-kozo. Taf. IV, Fig. rz.
Knospen wie bei der vorgenannten Art, verhaltnissmassig
gross, an die Triebe angedriickt. Schuppen dunkelbraun.
Blattpolster vorspringend ; Blattnarbe rund. Zweige schlank,
hin- u. hergebogen, dunkelbraun od. schwarzlichbraun. Lenti-
cellen sehr deutlich. Mark rund.

Broussontia papyrifera, Veni. Kajinoki. Taf. IV, Fig. 3.
Endknospen grosser als Seitenknospen, die ersteren locker
beschuppt, die letzteren dicht, von etwas kegeliger Form.
Zweige ziemlich dick, dunkelgraulich griin. Die jiingeren
Theile mit weissglanzenden, langen Haaren bedeckt. Blatt-

narbe gross, rundlich. Lenticellen deutlich. Mark weit.

?.—Blattnarbe nicht rundlich.
* —Zweige diinn.
Stillingia sebifera, Boxb. Nankin-haze.
Knospen klein, keilformig, an der Langtriebe sehr zahlreich
sitzend. Schuppen duukel od. schwarzbraun. Beiderseits der
Knospe je eine Nebenknospen sitzend. Jiinge Zweige, griin,

schlank, lang gestreckt; altere griinbraun. Blattnarbe halb-
mondformig. Lenticellen deutlich. Mark eng, rund.

Lagerstroemia indica, L. Sarusuberi. Taf. IV, Fig. 5.
Knospen keilformig, zugespitzt; Schuppen dunkelbraun.
Zweige diinn, braun gefarbt. Die ausseren Paridermschichten
reissen in Langsstreifen auf, und blattern sich in langen unre-
gelmassigen Bandern ab. Blattnarbe von Linsenform. Lenti-
cellen undeutlich. Mark rund. Die Knospen sind an den
Langtrieben zumeist gegenstandig angeordnet.

Spiraea betulifolia, Pall. Maruba-shimotsuke. Taf. IV, Fig. 6.

Knospen flach, die Spitze pfriemenformig, lang gestreckt,

u. gekriimmt, dunkelbraun. Zweige schlank, besenformig,

dunkelbraun. Blattpolster vorspringend. Blattnarbe sehr
klein. Mark weit, etwas eckig.

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 245

** —Zweige ziemlich dick.
Cladrastis amurensis, Benth. et Hook. var. floribunda, Max.

Inu-enju. Taf. IV, Fig. 7.
Knospen dicht. sitzend iiber der Blattnarbe, keilformig, am
Riicken derselben etwas nach vorne gewolbt; die Spitze
schwach gekriimmt. Schuppen schwarzbraun, wenig behaart.
Zweige dick, schwarz od. dunkelbraun. Blattnarbe sichelfor-
mig. Mark weit, von unregelmdssiger Form. Die Rinde hat
einen eigenthtimlichen unangenehmen Geruch. Das Holz

nimmt hellgelbe Farbung an.

Cereis chinensis, Bunge. Hanazuo. Taf. IV, Fig. 9g, ro.

Knospen klein, flach, keilformig, an der dem Triebe zuge-
wendten Seite abgeplattet, riickseits derselben eine kleine
Nebenknospe. Die Schuppen schwarzbraun. Blattnarbe vor-
springend, umfasst die Nebenknospe. Bliithenknospen kiefer-
zapfenformig. Zweige hellbraun, hin- u. hergebogen, mit
feinen Lenticellen versehen. Mark eng, rundlich.

Excoecaria japonica, #. Mucll. Shiraki. Taf. IV, Fig. 8.
Knospen keilf6rmig, zugespitzt. Knospenschuppen braun,
unterseits derselben Carminviolett gefleckt. Zweige grau-
braun, hin- u. hergebogen. Blattnarbe halbmondformig. Lent.

deutlich. Mark weit, rundlich. Die Rinde hat einen eigen-
thiimlichen Geruch.

Koelreuteria paniculata, Laxm. Mokugenju. Taf. IV, Fig. ¢.
Knospen ziemlich dick, von kugeliger Form. Schuppen
glanzend graubraun, behaart. Zweige dicker als vorgenannte
Arten, in der Langsrichtung 2-4 tiefe Furchen laufend, grau-
braun schwarzlich bestaéubt, schmutzig. Blattnarbe herzfor-
mig. Mark sehr weit, von unregelmassiger Form.

VII.—Knospen sitzend, von mehreren Schuppen
umgeben. Zweige auffallend schlank, od. besen-
formig.

A.—Zweige rothbraun, glanzend.
a.—Strauchartig.
Prunus japonica, Thunb. Niwa-ume. Taf. VI, Pig. 8.

Knospen klein, etwas kantig, réthlich, meist zwei derselben

246 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

nebeneinander sitzend. Zweige schlank, gerade, besenformig,
dunkelrotlich od. rothbraun, schmutzig bestaéubt. Blattpolster
ziemlich hoch.

Itea japonica, Oliv. Zuina. Taf. III, Fig. 20.
Knospen halbkugelig, von drei Schuppen (zu beiden Seiten

u. Riickwarts) umgeben. Zweige gelblichrothbraun. Blatt-
polster vorspringend. Lenticellen deutlich. Mark etwas eckig.

Spiraea cantoniensis, Louwr. Kodemari. Taf. IV, Fig. ro.
Knospen spindelformig, kurz, schwach eckig, hellbraun.
Zweige dunkelrothbraun, glanzend. Die ausseren Korkschich-
ten leicht abschalbar. Lenticellen undeutlich. Mark weit,
eckig.

b.—Baumartig.

Betula alba, L. subsp. verrucosa var. vulgaris, Regel. Shirakaba.
Taf. IV, Ftg.ce:
Knospen spindelig, lang. Schuppen dunkelgelbbraun, harzig.
Einjahrige Zweige gelbbraun, hin- u. hergebogen, kleinwalzig
mit einem weissen Ueberzug, sehr rauh anzufiihlen; die
alteren dunkelgelbbraun. Lent. deutlich. Mark sehr eng.
von unregelmassiger Form, dreieckig od. viereckig.
Betula Bhojpattra, Wall. var. typica. Regel. Onoore.
Taf. 1V, Pigs wees
Knospen eiformig, lang, locker beschuppt, behaart, hell-
braun teilweise dunkel gefarbt. Zweige schlank, kleinwalzig,
mit weisslichem Ueberzug, die jiingeren hellbraun, 4ltere
dunkelbraun Blattnarbe schmal. Lent. deutlich. Mark eng.
Das Holz dieser Art ist das harteste unter allen Betulaarten;
altere Stamme mit rauher Borke.
Betula alba, L. subsp. papyrifera var. communis, Rege/.
Makamba. Taf. IV, Fig. 415.
Knospen u. Zweige ahnlich wie bei der B. vulgaris, u. von
dieser unterschieden, durch weissbraune Rinde, wahrend B.
vulgaris silberweisse Rinde besitzt.
Betula globispica, Shivat. Jizokamba. Taf. IV, Fig. 16.
Knospen eiférmig, zugespitzt. Knospenschuppen dunkel-
braun od. hellbraun. Zweige schlank, graubraun. Blatt-

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 247

narbe halbmondformig. Lenticellen deutlich, linsenformig.
Mark schmal von unregelmassiger Form.

Betula alba, subsp. latifolia, var. Tauschii,? Taf. IV, Fig. rz.

Blattnarbe kurze, stumpfe Dreiecke. Lent. deutlich. Mark
eng, von unregelmassiger Form.

Amelanchier asiatica, Koch. Zaifuri-boku. Taf. IV, ig. 77.

Die ganze Form der Knospe eiformig, lang, sehr locker be-
schuppt. Die dussere Halfte der Schuppen u. die inneren
Schuppen pfriemenformig, hellroth gefarbt, am Rande dersel-
ben baumwolleartig lang behaart. Zweige schlank, roth od.
rothbraun glanzend. Die alteren Zweige dunkelgrau mit
mehreren gebriimmten Kurztrieben. Blattnarbe klein. Lent.
deutlich. Mark rund, weit.

B.—Zweige grau, dunkelgrau od. graubraun.

Pourthiana villosa, Decne. Ushikoroshi. Taf. IV, Fig. 78.

Knospen kegelig, ziemlich kurz, flach, 3-4 Schuppen
erkennbar, rothbraun, weiss gefleckt. Blattpolster vorsprin-
gend; Blattnarbe schmal. Zweige braunlich grau, schlank.
Lent. deutlich.

Stephanandra flexuosa, Sieb. et Zucc. Kogome-utsugi.
Taf: IV, Fig. 20.
Knospen spindelig, kurz, rothlich gefarbt. Zweige schlank,
lang gestreckt, hin- u. hergebogen, graubraun, etwas glanzend.
Die Zweige an der Haupttriebe fast senkrecht stehend. Lent.
undeutlich. Mark weit, etwas gefarbt.

Spiraea japonica, L. fil. Shimotsuke. Taf. IV, Fig. 2z.
Knospen kegelformig, kurz, von mehreren pfriemformigen
Schuppen umgeben, hellbraun. Zweige graubraun od. braun,
kleinwalzig. Lent. undeutlich. Mark sehr weit.

Spiraea prunifolia, Sieb. et Zucc. Shijimibana.

Knospen spindelig, kurz. Zweige graubraun, schlank, besen-
formig. Mark weit, rund.

248 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

VIII.—Knospen sitzend, von mehreren Schuppen
umgeben; einjahrige Zweige aufallend dick.
A.—Zweige mit Stacheln.
Aralia sinensis, 2. Tara. Taf. IV, Fig. 22.

Knospen kegelig, locker beschuppt. Endknospe grésser als
Seitenknospen. Schuppen graubraun, od. schwarzgraubraun.
Die jungen Zweige graubraun, behaart, wie mit Baumwolle
bedeckt. Zahlreiche, weiche Stacheln auf der ganzen Ober-
flache der Zweige. Blattnarbe schmal, u. lang, umfasst bei-
nahe die ganze Triebe. Mark sehr weit, rund. Die Rinde
harzig. Bei Verwundung fleisst eine durchsichtige Fliissig-
keit aus.

Acanthopanax ricinifolium, S7cb. et Zucc. Hari-giri.
Taf. "Vs rigeaean
Endknospe grésser als Seitenknospen, halbkugelig, etwas
zugespitzt, od. kegelig, am Triebende dicht stehend. Sei-
tenknospen keilformig. Schuppen derb, dunkelcarminroth,
glanzend. Zweige gelbgriin, teilweise grauweiss glanzend.
Zahlreiche, derbe gerade Stacheln an der ganzen Oberflache
der Zweige. Blattnarbe schmal, umfasst die Knospe. Mark
weit, rundlich.

B.—Zweige ohne Stacheln.

a.—Knospen lang gestreckt, rothlich od. braun gefarbt..
Pirus sambucifolia, Cham. et Sch. Nanakamado. Taf. V, Fig. 2.
Knospen spindelig, lang. Endknospe grosser als Seiten-
knospen, die letzteren an die Zweige angedriickt, u. an der
Spitze etwas gebogen. Schuppen dunkelroth glanzend. Die
Zweige auf der Lichtseite braun, auf der Schattenseite grau-
braun, glanzend. Lent. gross, deutlich. Blattnarbe sehr
schmal. Mark rund. Die Rinde hat einen eigenthiimlichen

Geruch.

b.—Knospen sitzend, griingelb od. grau gefarbt.

Acanthopanax sciadophylloides, Fr. et Sav. Gonzetsu. .
Taf. V, Fig. 3, ¢.

Knospen an der Spitze der Triebe dicht sitzend, kegel-
formig, kurz. Schuppen griingelb. Junge Zweige gerade,

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 249

grau gefarbt, mit grossen deutlichen Lenticellen versehen ;
die alteren grauweiss, dunkelblau gefleckt. Kurztriebe sehen
wie grosse Seidenwiirmer aus. Blattnarbe schmal u. lang.
Mark gefachert. Die Rinde hat einen Geruch.

IX-—Knospen sitzend von mehreren Schuppen
umgeben. Die Knospen stehen an der Spitze der
Langtriebe einzeln od. an den Kurztrieben
einzeln od. gehauft.

A.—Zweige mit Stacheln, d. h. mit metamorphosirten
Blatt- u. Haargebilden; dornspitzige Zweige nicht
vorhanden.

a.—Je Zwei gerade Stacheln neben den Knospen.

b.—Gerade einfache od. verzweigte 3~5 spitzige Stacheln
unter den Knospen.

Ribes grossularia, L. Gusuberii.

Knospen schief abstehend, spindelig, schlank, von diinnen
zugespitzten od. ausgefransten Knospenschuppen, nicht je-
doch von Resten der Blattbasis umgeben. Die einjahrigen
Zweige sind hellgrau, die Oberhaut springt in Langsrissen
auf, u. lasst darunter die graubraune, od. dunkelcarmine,
glatte Rinde erkennen. Die Stacheln unter den Knospen
meist zu drei beisammenstehend, seltner ist bloss ein Stachel
vorhanden. Ausserdem koénnen aber auch noch Stacheln auf
der ganzen Oberflache der Zweige vorkommen.

Berberis vulgaris, L. Hebinoborazu. Taf. V, Fig. 6.

Die Knospen von der stehenbleibenden Basis mehrerer
Blatter umgeben. Die braunen Knospenschuppen sehen wie
vertrocknet aus. Zweige lang ruthenformig, dunkelbraunge-
farbt. Die Rinde nach dem Aufschneiden intensiv gelb ge-
farbt. Stacheln an der Spitze der Triebe in der Einzahl, an
der tibrigen Theilen verzweigt mit 3, 4 od. 5 Spitzen. Stacheln
nur unter den Knospen. Mark ziemlich weit, gelblich gefarbt.

Berberis Thunbergii, DC. Megi. Taf. V, Fig. 5.

Die Knospen sind kugelig, locker beschuppt. Die schuppen
braun. Die zahlreichen feinen Zweige tibereinander hervor-
tretend, dunkelrothbraun od. graubraun gefarbt. Die Stacheln

250 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

in der Einzahl, u. unterseits der Knospen. Die Rinde, Mark
u. das Holz intensiv gelb gefarbt. Mark ziemlich weit.

c.—Meist Zwei od. nur eine noch riickwarts gebogene,
derbe Stachel unter jeder Knospe.
Rosa multiflora, Thunb. No-ibara.
Knospen eiformig, zugespitzt ; die Schuppen glanzend roth.
Die Zweige auf der Sonnenseite dunkelroth, auf der Schatten-
seite griin. Je zwei Stacheln unter jeder Knospe einige mm.
entfernt.. Lent. wenig. Mark weit.

Acanthopanax spinosum, Wig. Ukogi.

Knospen klein, halbkugelig, dicht sitzend. Zwei dussere,
grossere, hellbraun gefarbte Schuppen umgeben die Knospe von
beiden Seiten. Zweige graubraun, theilweise schwarz gefarbt,
od. grau. Stacheln meist einzeln. Lent. gross, deutlich.
Mark sehr weit. Rinde hat einen eigenthtimlichen Geruch.

d.—Zahlreiche derbe, zurtickgebogene Stacheln, ohne
Zusammenhang mit den Knospen.
Rubus incisus, Thunb. Ki-ichigo.

Knospen eiformig, zugespitzt, dunkelréthlich, teilweise
schwarz. Zweige gerade, dunkelroth gefarbt, bereift. Blatt-
narbe schmal. Mark sehr weit.

B.—Zweige ohne Stacheln, mit od. ohne dornige Zweige.
a.—Knospenschuppen griin, teilweise braun gerandert.
a.—Knospen, besonders die unteren Seitenknospen
klein.
Ficus erecta, Thunb. Inu-biwa. Taf. V, Fig. 9, Zo.

Endknospe wesentlich grésser als Seitenknospen, spindel-od.
kegelformig. Seitenknospen spindelig, od. kugelig von mehre-
ren Schuppen lose umgeben. Zweige griin. Blattnarbe gross,
rundlich. Lent. wenig, deutlich. Mark weit.

Helwingia ruscifolia, Willd. Hanaikada.
Unter VI, B. b. #. beschrieben.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 251

£.—Knospen gross, od- ziemlich gross.
1.—Knospen kugelig od. eiformig.
Spiraea Thunbergii, Sicb. Hozaki-nanakamado.
DG NARS AT, Ls
_ Knospen locker beschuppt. Zweige weisslich girau glanzend.
Blattnarbe gross von unregelmassiger Form, gewolbt. Lent.
wenig, deutlich. Mark sehr weit, hellbraun gefarbt.

2.—Knospen lang gestreckt.
Populus suaveolens, isch. Dero. Taf. V, Fig. 73.

Knospen spindeliormig lang, etwas kantig. Die Seitenknos-
pen an der Spitze gekriimmt. Die Schuppen griin, theilweise
braun, harzig, glanzend. Zweige graugriin od. graubraungriin,
glanzend. Blattnarbe sichelformig, schmal. Lent. wenig.
Mark strahlenformig, griinlich gefarbt.

b.—Knospen harzig, glanzend braun.

Populus tremula, L. var. villosa, Wesm. Yamanarashi.
Taf. V, Fug. 14.
Knospen spindelig, kurz, gerade od. nach innen gebogen.
Die Schuppen braun glanzend, teilweise mit Harz bedeckt.
Die jungen Zweige braun od. rothbraun, glanzend od. behaart ;
die alteren grau. Blattnarbe sichelformig. Lent. deutlich.
Mark fiinfeckig.

Pirus aria, Ehvh. var. Kamaonensis, Wall. Urajironoki.
Taj, V, Pig. 27.
Knospen kegelig, (die Seitenknospen etwas abgeplattet),
kurz, von mehreren Schuppen lose umhiillt. Die Schuppen
dunkelbraun, glanzend od. gelbgriinlich. Zweige rothbraun,
die alteren mit zahlreichen Kurztrieben. Blattnarbe sichelfor-
mig, schmal. Lenticellen deutlich. Mark weit, rundlich.

Betula grossa, Sieb. et Zucc. Mizume. Taf. V, Fig. 27.
Knospen eiformig zugespitzt, in der Mitte etwas hervor-
ragend. Die Schuppen gelblichbraun, dunkelrothbraun geran-
dert, glanzend. Am Rande derselben ein wenig behaart. Hin-
jahrige Zweige hell- od. graulichrothbraun, glanzend, schwach
behaart, die alteren dunkelrothbraun. Mark eng, eckig. Die
Rinde hat einen eigenthiimlichen Geruch.

252 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

<

Betula ulmifolia, S. ef Z. Joguso-minebari. Taf. V, Fig. 15, 16.

Knospen ziemlich lang, spindelig. Die Schuppen dunkel-

rothbraun, glanzend. Blattnarbe flachdreieckig. Die Rinde
hat einen eigenthtimlichen Geruch.

Betula Maximowicziana, Regel. Saihada. Taf. V, Fig. 78.
Knospen ciférmig, zugespitzt od. spindelig. Die Schuppen
gelbbraun, dunkel gerandert, glanzend, harzig. Einjahrige
Zweige schwach fein behaart, dunkelgelblichbraun, glanzend.
Die rundlichen od. elliptischen Lenticellen sind deutlich.

Mark sehr eng, eckig.

c.—Knospen roth, hell- od. dunkelbraun, od. graubraun,
behaart od. nicht.

a.—Dornspitzige Zweige sind vorhanden.

*,—Knospen behaart.

Pirus japonica, Thunb. Boke. Taf. V, Fig. 22.

Knospen kegelférmig, kurz, flach, u. klein. Die Schuppen
dunkelbraun behaart ; die Zahl der Schuppen ist gering. Ein-
jahrige Zweige braun bis griinbraun, die alteren dunkel- od.
graubraun. Blattmarbe dreieckig. Lenticellen wenig. Mark

- rundlich.

Pirus sinensis, Lindl. Nashi. Taf. V, Fig. 27.

Endknospen meist grdsser als Seitenknospen, kegelig. Die
Schuppen dunkelrothbraun, behaart od. wenig behaart, weiss-
lich bereift. Zweige griinlich braun, gerade. Blattnarbe
dreieckig, schmal. Lent. ganz deutlich. Mark ziemlich weit,

fiinfeckig.

** —Knospen nicht behaart.

Mespilus cuneata, Sicb. ef Zucc. Sanzashi. Taf. V, Fig. 23, 24.
Knospen kugelig klein, an der Triebe fast senkrecht liegend,
rothbraun, glanzend, grauweissgerandert. Schuppen zahl-
reich, aber etwas undeutlich. Junge Zweige braun, glanzend,
altere grau. Die Verzweigung dicht, die einzelnen Zweige

von Zicksackform.

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 253

Glochidion obovatum, Sieb. ef Zucc. Kankonoki.
Rape Vi, bigs Tr.
Knospen kugelig locker beschuppt, graubraun, sehr wenig
behaart. Zweige hellbraun, vertrocknet aussehend. Die dor-
nigen Zweige schwach. Lent. wenig. Mark ziemlich weit,
eckig.

Prunus Mume, Sieb. et Zucc. Mume.

Knospen kegelformig, kurz, dunkelréthlich gefarbt. Die
jiingen Zweige griin, teilweisse rothlich, altere grau od. grau-
lichbraun. Blattpolster hoch. Blattnarbe halbmondformig.
Mark weit, etwas eckig. Im Winter sind kugelige Blititen-
knospen vorhanden.

8.—Dornspitzige Zweige nicht vorhanden.
t.—Blattnarbe halbmondformig, schmal.
a.—Knospen behaart.
Lindera glauca, Bl. Yama-kobashi. Taf. VI, Fig. 2.

Knospen spindelig, ziemlich kurz, u. dick. Die Schuppen
hellrothbraun, die inneren behaart, die dusseren nicht. Blatt-
narben halbkreisformig. Die jiingen Zweige graubraun, glan-
zend. Lenticellen fein, wenig deutlich. Mark rund. Die
Rinde hat einen eigenthiimlichen Geruch.

Lindera umbellata, Thunb. Kanakuginoki. Taf. VI, Fig. 6.

Knospen spindelformig, locker beschuppt, dunkelrothbraun,
theilweise hellbraun gefarbt, schwach behaart. Blattnarbe
halbmondformig od. etwas rundlich. Die Rinde hat einen
eigenttimlichen Geruch.

Prunus persica, Sieh. et. Zucc. Momo.
Knospen eiformig griinlich, filzig behaart. Zweige griin
teilweise dunkelréthlich. Blattpolster vorspringend. Blatt-
narbe etwas klein, halbkreisformig. Mark eckig (sechs).

Prunus tomentosa, Thunb. Yusuraume. Taf. VI, Fig. 9.

Knospen mit pfriemformiger Schuppen neben der Basis,
kegelformig, zugespitzt, ziemlich lang, (Endknospen grésser
als Seitenknospen, locker beschuppt). Seitenknospen von den
Langtrieben abstehend. Die Schuppen dunkelrothlich, behaart.

254 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Die jungen Zweige braun od. graulichbraun, dichtfilzig be-
haart. Mark klein, rund.

Liquidambar Maximowiczii, Mig. Fu. Taf. VI, Fig. 7.
Endknospen grésser als die Seitenknospen, spindelig. Die
Schuppen carminrothbraun, teilweise hellbraun gerdandert,

seidenglanzend, schwach behaart. Zweige hellbraun, wenig
behaart. Blattnarbe halbmondformig, schmal. Mark eckig.

b.—Knospen unbehaart.
Pirus cathayensis, Heml. Kuarin. Taf. VI, Fig. ro.

Endknospen etwas kugelig, Seitenknospen dreieckig, gegen
Triebe zu abgeplattet. Zwei grosse, glanzend braune Schup-
pen umgeben fast die ganze Knospe. Die einjahrigen Zweige
hellbraun, glanzend, schwach behaart, die alteren dunkel-
braun. Blattpolster hoch. Blattnarbe sichelformig. Zahl-
reiche Kurztriebe sind vorhanden, fast gleich lang. Lent.
wenig. Mark rund.

Orixa japonica, Thunb. Kokusagi. Taf. VI, Fig. rz.

Knospen zugespitzt, vierkantig, auf allen Vierseiten von
Knospenschuppen umgeben, glanzend dunkelrothbraun ge-
farbt. Zweige graugriin, glanzend, auf der Schattenseite
griin, auf der Lichtseite etwas dunkel. Blattnarbe halbmond-
formig. Lent. deutlich. Mark von unregelmassiger Form.
Die Rinde hat einen eigenthiimlichen, unangenehmen Geruch.

Disanthus cercidifolia, Max. Beni-mansaku. Taf. VI, Fig. rz.

Knospen spindelig etwas geplattet. Mehrere glanzend
dunkelrothen Schuppen umgeben, abwechselnd die Knospen
von beiden Seiten. Blattnarbe halbmondformig. Zweige
hin- u. hergebogen, auf der Lichtseite dunkelrothlich. Lent.
zahlreich, deutlich. Mark schmal, rundlich.

Kerria japonica, DC. Yamabuki.

Knospen spindelformig, rothbraun, die Spitze etwas gebogen.
Zweige griin, hin- u. hergebogen. Blattnarbe halbkreisformig,
schmal. Lent. deutlich. Mark sehr weit.

Lindera triloba, B/. Shiromoji. Taf. VI, Fig. 3.

Knospen mit kugeligen Bliitenknospen an der Basis, spin-

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 255

delig, gelblich od. griinlichbraun, dunkel gerandert. Zweige
hin- u. hergebogen, auf der Lichtseite dunkelrothbraun, auf
der Schattenseite griinlich gelb, die alteren dunkelgrau.
Blattnarbe halbmondformig. Mark weit. Die Rinde hat

eigenthiimlichen Geruch.

2.—Blattnarbe rundlich.

*,.—Knospen kegel od. spindelformig, ziemlich gross.
Lindera obutusiloba, B/. Dankobai. Taf. VI, Fig. 73.

Die Knospen meist mit kleinen Nebenknospen, rothbraun,
glanzend, spindelig, die Spitze etwas gebogen. Die jungen
Zweige auf der Lichtseite rothbraun, sonst gelblich griin, die
alteren grau. Lenticellen deutlich. Mark sehr weit.

Lindera praecox, B/. Aburachian. Taf. VI, Fig. 74, 75.

Knospen lang gestreckt, spindelig. Die Seitenknospen nach
innen etwas gebogen. Die Seitenknospen dunkelrothbraun.
Die “‘jungen Zweige braunlich grau glanzend wie lackiert.
Lent. ziemlich deutlich. Die Rinde hat einen eigenthiimlichen
Geruch.

**.—Knospen sehr klein, kugelig.
3.—Blattnarbe dreieckig, schmal.
a.—Knospen behaart.
Prunus Miqueliana, Maxim. MHigan-zakura.

Lape Vai FO 27;

Knospen spindelformig, klein, an der Langtriebe gehauft,

od. auch nicht. Schuppen dunkelbraun, behaart. Blattpolster
etwas vorspringend. Zweige schlank, die einjahrige hellbraun

behaart, die dlteren schwarzbraun. Lent. deutlich. Mark
rund.

Pirus Toringo, Sicb., var. incisa, Fv. et Sav. Hime-kaido.
Taf Ee. 255120.
Knospen an der Langtriebe kegelformig, kurz, dreikantig.
Seitenknospen flach, an die Zweige gedriickt. Gewéhnlich
2-4 Schuppen sichtbar, glaénzend rothbraun, weisslich bereift.
Zweige dunkelréthlich braun, glanzend. Die Seitenzweige
aus den Langtrieben fast senkrecht hervortretend, kurz, dorn-

256 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

spitzig. Mark rund. Nach Abschneiden der Zweige nimmt
die Rinde u. das Mark eine gelblichbraune Farbung an.

b.—Knospen unbehaart.

Prunus Grayana, Max. Uwamizu-zakura. Taf. VI, Fig. 79, 20.

Die Endknospe kugelig, die Seitenknospen etwas flach,
dunkelrothbraun gefarbt, glanzend. Blattpolster kugelig her-
vorgewolbt. Zweige ziemlich dick, die jiingeren dunkelbraun,
teilweise weisslich, die alteren schwarzbraun, glanzend. Lent.
deutlich. Mark rund. Die Rinde hat einen eigenthiimlichen
Geruch.

Prunus Buergeriana, Mig. Inu-zakura. Taf. VI, Fig. 78.

Endknospe kegelig, kurz, Seitenknospe etwas kugelig od.
zugespitzt. Gewodhnlich 4 Schuppen sichtbar, dunkelroth-
braun glanzend. Blattpolster ziemlich hoch. Zweige dunkel-
braun, Stamm graubraun gefarbt. Mark rund, etwas gefarbt
Die Rinde hat einen eigenthiimlichen Geruch, starker als die
vorhergenannte Art.

Prunus Maximowiczii, Rupy. Mejiro-zakura. Taf. VI, Fig. 23.
Endknospen cylindrich zugespitzt ; Seitenknospen eiformig
gespitzt ; die Spitze schwach behaart. 6-7 Schuppen sichtbar,
locker beschuppt, dunkelbraun, weisslich gefleckt. Junge
Zweige grau, schmutzig, altere glanzend dunkelbraun. Lent.
deutlich. Mark etwas eckig.

Prunus cerasoides, Max. Choji-zakura. Taf. VI, Fig. 27, 22.

Knospen an der Langtriebe gehauft, spindelformig, kurz,
hellbraun od, dunkelbraun glanzend. Zweige hin- u. hergebo-
gen, kurz. Die jiingeren Zweige glanzend braunlichgrau,
teilweise weisslich, die alteren hellbraun. Lent. deutlich.
Mark rund.

Prunus pseudocerasus, Lindl. var. spontanea, Maxim.
Yama-zakura. Taf. VI, Fig. 24.

Knospen spindelig, an der Langtriebe gehauft od. auch nicht.
Die Schuppen dunkel- od. hellbraun, 6-8 sichtbar, locker
beschuppt. Junge Zweige graubraun, 4ltere dunkelbraun.
Mark 5 eckig.

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 257

Pyrus Miyabei, Sargent. Azuki-nashi. Taf. V, Fig. rg, 20.
Knospen eiformig zugespitzt. Schuppen glanzend dunkel-
roth, am Rande braun, schwach behaart. Junge Zweige
braun, altere dunkelbraun glanzend. Kurztriebe eng geringelt.
Lenticellen weissgrau, sehr deutlich. Mark ziemlich weit ;

von unregelmassiger Form.

Rubus trifidus, Thunb. Kayji-ichigo.
Knospen ziemlich gross, spindelig, etwas gekriimmt. Schup-
pen dunkelroth, am Rande schwach behaart. Zweige hin- u.
hergebogen, auf der Lichtseite dunkelroth, glanzend, auf der
Schattenseite griin. Blattnarbe dreieckig, schmal od. sichel-
formig. Mark sehr gross, etwas gefarbt.

4.—Blattnarbe gross, herzformig,

Platyearya strobilaceae, Sich. et Zucc. Nobunoki.
Tap WV ic. 25.
Endknospe grosser als Seitenknospe, zugespitzt, rothbraun,
dunkelbraun geradndert. Zweige dick, graubraun, die jiingeren
behaart, die alteren glanzend. Lent. fein, zahlreich. Mark
weit, von unregelmassiger Form.

Euptelaea polyandra, Sich. et Zucc. Fusa-zakura.
Way Mill) ie, 2.
Knospen eiformig zugespitzt. Schuppen glanzend dunkel-
violettbraun. Einige an der Basis der Knospe grauweiss be-
haart. Blattnarbe gross, umfasst beinahe die ganze Knospe.
Junge Zweige braun, altere graubraun. Lent. deutlich. Mark
ziemlich eng. Zweige meist zweizeilig angeordnet.

X.—Knospen sitzend von mehreren Schuppen umgeben,
an der Spitze der Langtriebe gehauft.
A.—Knospen nicht von pfriemenformigen Nebenblatt-
chen umgeben.
Quercus glandulifera, B/. Konara. Taf. VII, Fig. 2.

Knospen zugespitzt, fiinfkantig, kahl, schief an der
Triebe. Schuppen dunkelbraun, (seltner sind pfriemenformige
Schuppen an der Basis der Knospe vorhanden). Blatt-

258 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

narbe flachdreieckig, etwas vorspringend. Zweige grau, auf
der Schattenseite hellbraun. Lent. ziemlich lang, hervor-
tretend, sehr deutlich. Mark strahlenformig.

Quercus grosseserrata, B/. Mizunara. Taf. VII, Fig. 3.
Knospen zugespitzt, undeutlich fiinfeckig, lang. Schuppen
hellbraun. Zweige griinlich graubraun glanzend. Die obere,
graue feine Haut leicht sich abschilfernd. Blattnarbe drei-
eckig od. halbmondformig. Lent. weniger deutlich als bei
Quercus glandulifera. Mark strahlenf6rmig.

Quercus serrata, var. variabilis, B/. Abemaki. Taf. Vil, Fig. 5.

Knospen kegelférmig, etwas kantig, wenig behaart (seltner
mit pfriemenformigen Schuppen versehen). Zweige grau, auf
der Schattenseite graubraun.

Die vertrockeneten Laubblatter den ganzen Winter hindurch
nicht abfallend. Lent. zahlreich, fein, breiter als lang. Das
Aussehen des Quercus variabilis ist ahnlich wie die Q. serrata,
aber die Knospenform ist wie bei der Q. glandulifera.

Quercus dentata, Thunb. Kashiwa. Taf. VII, Fig. 4.

Endknospe grésser als Seitenknospen, od. als die der vor-
gesetzten Arten, kegelig, fiinfkantig. Seitenknospen etwas
lang, die Spitze gebogen, iiber der Blattnarbe anfrecht
stehend. Schuppen filzig behaart. Zweige dick, schwarzlich-
grau bestaubt, schmutzig. Das diirre Laub bleibt den ganzen

Winter an den Zweigen. Mark strahlenformig.

B.—Endknospen von pfriemenférmigen Nebenblattchen
umgeben.

Quercus serrata, Thynb. Kunugi. Taf. VII, Fig. 6.
Seitenknospen iiber der Blattnarbe stehend, einige derselben
mit Nebenknospen. Zweige graubraun dichtfilzig behaart,
mit Langsfurchen. Knospenschuppen graubraun, weniger be-
haart. Blattnarbe halbmondférmig, etwas vorspringend.
Lent. deutlich. Mark strahlenformig. Das diirre Laub bleibt

den ganzen Winter hindurch.

e
SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 259

XI.—Zweige kletternd, (Schlinggewachse).
1.—Knospen sind in der Blattachsel verborgen.
Actinidia arguta, Planch. Shira-kuchi. Taf. VII, Fig. 7.
Knospen hinter der Blattachsel verborgen, u. aus der
hiigeligen W6lbung des Zweiges hervortragend. Blattnarbe
halbkreisformig od. rundlich. Die jungen Zweige hell- od.
dunkelbraun, die dlteren graubraun, mit kiirzeren Seiten-
zweige. Lenticellen lang, viel, deutlich. Mark gefachert,
etwas gefarbt.

Actinidia polygama, Mig. Matatabi. Taf. VII, Fig. 8, 9.

Die Stelle der Knospen wie bei der vorigen Art. Blattnarbe
halb-elliptisch, tief gewolbt. Junge Zweige schlank, langge-
streckt, dunkelrothbraun glanzend. Lent. deutlich, etwas
kurz. Mark gefachert, gefarbt.

Menispermum davuricum, DC. Komorti-kazura.
Taf. VII, Fig. ro.
Knospen sind in der Blattnarbe verborgen, od. zwischen den
Kliiften sitzend, welche durch Rindenrisse an den Blatt-
achseln entstehen. Zweige glanzend dunkelrothbraun. Blatt-
narbe gross, rundlich od. herzformig, gewolbt. Lent. wenig
zahlreich, deutlich. Mark weit, rundlich.

2.—Knospen sind von unausgebildeten od. zusammen-
gedriickten Blattchen umgeben,
a.—Von unausgebildeten Blattchen.
Rhus toxicodendron, L. var. radicans, Mig. Tsuta-urushi.
Dafa Djs Big.: 75.
Ein grosses, hellbraunfilzig behaartes, unausgebildeten Blatt-
chen neben der Endknospe stehend. Seitenknospen flach,
kleiner. Zweige graulichrothbraun mit vielen deutlichen
Lenticellen, die alteren Zweige mit Luftwurzel versehen.
Blattnarbe glanzend rothbraun, gefarbt. Mark weit, rundlich.

b.—-Von zusammengedriickten Blattchen.
Coculus Thunbergii, DC. Ao-tsuzurafuji. Taf. VII, Fig. rz.

Knospen tiber der Blattnarbe sitzend. Die Zahl der zusam-
mengedriickten Blattchen ganz undeutlich, dunkelgrau, dicht-

260 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

filzig behaart. Zweige schlank, griin- od. dunkelgriinlich
braun, schwach behaart. Blattnarbe rundlich, gewolbt.

3-—Knospen mit einer bezw. zwei Schuppen umgeben.

A.—Zweige ohne Ranken.

Berchemia racemosa, Sich. et Zucc. Kuma-yanagi.
Taf. Vil, tig. ee go-
Knospen tiber dem Blattpolster stehend, nur eine Schuppe
umhiillt dieselbe vollstandig von der Riickseite aus. Schuppen
u. Zweige glanzend dunkelrothbraun, glatt. Lent. undeutlich.
Mark rund, ziemlich weit.

Wistaria chinensis, Sich. cf Zucc. Murasaki-fuji.
Taj. VII, fg. Fe:
Knospen kegelformig, gegen die Triebe zu abgeplattet.
die Spitze etwas gekriimmt. Schuppen dunkelréthlichbraun,
Zweige grau, glanzend. Blattpolster hochspringend, seitwarts
mit kleinem Rindenansatz. Blattnarbe rundlich, od. halb-
mondformig. Lent. deutlich. Mark weit, rundlich.

Wistaria brachybotrys, Sieb. ef Zucc. Shiro-fuji.
Taf. VII; Hig zee.
Knospen eifo6rmig, zugespitzt, dunkelrothbraun. Blattnarbe
halbmondformig. Zweige dunkelbraun. Lent. undeutlich.
Mark rundlich, weit.

B.—Zweige mit Ranken.

Vitis vinifera, L. Budo. Taf. VII, Fig. 77.

Knospen eiformig. Zwei Knospen-Schuppen, die innere
umhiillt fast die ganze Knospe, u. die dussere steht seitwarts ;
die letztere dunkelbraun gefarbt. Zweige von derselben Farbe,
hin- u. hergebogen. Lent. undeutlich. Mark rund, etwas
gefarbt.

Vitis flexuosa, Thunb. Gydjanomizu. Taf. VII, Fig. 78.

Knospen eifotmig. Zweige schlank, graubraun glatt ; Knos-
penschuppen von derselben Farbe. Blattnarbe undeutlich
dreieckig. Mark rund, eng.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 261

Vitis Thunbergii, Sieb. et Zucc. Ebizuru.
Knospen u. Zweige wie bei der vorigen Arten, aber davon
leicht zu unterscheiden, durch die baumwolleartige Behaarung,
welche die ganze Flache der Zweige umfasst.

4.—Knospen von mehreren Schuppen umgeben.
a.—Knospen spindelformig.

Schizandra chinensis, Baill. Chosen-gomishi.
Lap Vids Beg. 19.
Knospen spindelig, von mehreren Schuppen spiralig um-
geben. Schuppen tiefrothbraun; Zweige von derselben Farbe
auf der Lichtseite graulich. Blattnarbe halbmondformig.
Lent. sehr deutlich. Mark weit, rund. Die Rinde hat einen
eigenthtimlichen, angenehmen Geruch.

£.—Knospen kugel- od. eiformig.
Akebia quinata, Decne. Akebi. Taf. VII, Fig. 20.

Knospen eiformig, zugespitzt. Zwei od. drei derselben
beisammensitzend, locker beschuppt, dunkelbraun. Zweige
dunkelbraun, die alteren graubraun, glanzend. Lent. sehr
deutlich, zahlreich. Mark eng.

Akebia lobata, Decne. Mitsuba-akebi. Taf. VII, Fig. az.

Knospen eiformig, dunkelbraun. Seitwarts der grésseren
Knospen, die kleineren beisammensitzend. Blattpolster
hoch. Zweige braun od. hellbraun, glanzend. Lent. deut-
lich, weniger zahlreich als bei Akebia quinata. Mark rund.

Celastrus articulatus, T/iwnb. Tsuru-umemodoki.
Taf. VII, Fig. 22.
Knospen kugelig, an der Triebe senkrecht stehend, dunkel-
braun gefarbt. Zweige, gerade, graubraun. Blattnarbe halb-
mondformig. Lent. klein, zahlreich. Mark rund.

262 SHIRASAWA!: LAUBHOLZER IM WINTERZUSTANDE.

TABELLE II.

Knospen stehen an der Langtrieben abwechselnd,
** Zweizeilig.,,

I.—Knospen (Laub od. Bliten) gestielt, u. von wenigen
Schuppen umgeben.
A.—Knospen kugelig od. eiformig.

Corylopsis pauciflora, Sieb. et Zucc. Hiuga-mizuki.

Taf. VIII, Bigex.

Bliitenknospen eiformig od. kugelig, gestielt. Die ausseren

Schuppen glanzend hellbraun u. leicht ablosbar, die inneren

auf der Lichtseite hellrothlich, sonst griin. Zweige schlank,

die jiingeren braun glanzend, die alteren graubraun. Lent.
deutlich. Blattnarbe klein, dreieckig. Mark eng.

B.—Knospen spindelig od. kegelformig.
Corylopsis spicata, Sieb. ef Zucc. ‘Tosa-mizuki.

Taf. Vill, Fig. 2.

Bliitenknospen lang gestreckt, spindelig. Die dusseren

Schuppen leicht ablosbar, die inneren auf der Lichtseite

rothlich, od. intensivroth glanzend, sehr schon, sonst gelbgriin.

Zweige hin- u. hergebogen, die jiingeren glanzend graubraun,

die alteren etwas dunkler. Blattnarbe ziemlich gross, drei-
eckig. Mark rund.

C.—Knospen etwas flach.

Stuartia monadelpha, Sieb. et Zucc. Saruta,
Taf. VIH, Fig. 3, 4.
Knospen etwas flach, von beiden Seiten beschuppt, scharf-
kantig, hellgrau, braun behaart. Zweige schlank, besen-
formig. Der Stamm braun, glanzend, glatt. Lent. deutlich.
Mark rund.

Stuartia pseudo-Camellia, Max. Natsu-tsubaki.
Taf. Vill, Fags.
Knospen spindelig, etwas abgeflacht. Die ausseren Knos-
penschuppen dunkelbraun, leicht ablosbar, die inneren

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 263

griingrau, filzig behaart. Zweige hin- u. hergebogen; die
Peridermschichten reissen faserig auf. Die Farbung der ein-
jahrigen u. mehrjahrigen Zweige deutlich verschieden. Mark
rund, etwas weit.

II.—Knospen kugelig, keil- od. eiformig, mit zwei od.
weinigen Schuppen.
A.—Knospen dusserlich von einer grossen umfassenden,
u. einer kleineren Schuppe umgeben.

Tilia cordata, Mzll. var. japonica, Mig. Shinanoki.
Taj. Vil, Fie. Se
Knospen eif6rmig zugespitzt, etwas gekriimmt, Endknospen
grosser als Seitenknospen u. von unregelmassiger Form,
dunkelrothlich, kahl. Zweige etwas gebogen, die einjahrigen
hellbraun, auf der Lichtseite réthlich, glanzend, die alteren
dunkelgrauroth. Blattnarbe halbmondformig. Mark schmal.
Tilia Miqueliana, Max. Bodaiju. Taf. VIII, Fig. 6, 7.
Knospen kugelig zugespitzt, zwei Seiten derselben abge-
flacht, gelbgriin schwach behaart, an der Triebe schief an-
biegend, u. gebogen. Einjahrigen Zweige gelbgriin, kurz
behaart, die alteren graubraun. Mark rund.

B.—Knospen von mehreren Schuppen umgeben.
a.—Knospen behaart.

Celtis sinensis, Pevs. Enoki. Taf. VIII, Fig. 9.

Knospen keilformig, flach an der Trieb angedriickt, mit
zwei pfriemenformigen Blattchen an der Basis. Zwei
Knospenschuppen sichtbar, schwarzlichgrau behaart. Die
einjahrigen Zweige dicht behaart, dunkelgraubraun. Blatt-
polster hoch. Lent. deutlich. Mark rund.

Corylus heterophylla, Fisch. Hashibami.

Taf, VU re. £2) 13%

Knospen eifoérmig, die vordere u. hintere Seite etwas abge-

flacht, dunkelrothbraun, am Rande schwach behaart, sonst

kahl. Die jiingere Triebe rothlichbraun, bereift. Im

Winter mit rothlichbraun gefairbten Bliitenkatzchen. Blatt-

polster hoch. Blattnarbe abgerundet dreieckig. Mark rund,
eng.

264 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

Corylus rostrata, Ait. var. Sieboldiana, Max.

Tsuno-hashibami. Taf. VIII, Fig. zz.

Knospen kugelig, zugespitzt. Die Spitze u. der Rand der

Schuppen behaart, dunkelroth gefarbt. Zweige grau, in den

ersten Jahren sehr dicht behaart. Katzchen groésser als bei

Corylus heterophylla 2.5-3.5 cm. lang, graubraun gefarbt.

Blattnarbe halbmondformig. Mark unregelmassiger Drei-
eckig, eng.

2.—Knospen nicht behaart.
Castanea vulgaris, L. var. japonica, DC. Kuri.
Taf. VIII, Fig: zz.
Knospen kugelig, zugespitzt, riickwarts etwas flach. Schup-
pen glanzend dunkelrothbraun. Zweige von derselben Farbe.
Blattnarbe dreieckig. Lenticellen, weiss, deutlich. Mark
strahlenformig. Der Holztheil der jiingeren Zweige nimmt eine

unregelmassige Form an.

III.—Knospen spitz, walzenformig, spindel- od. kugel-
formig, mit zahlreichen Schuppen.

A.—Schuppen spiralig angeordnet.
a.—Knospen spindelig.
Fagus sylvatica L. var. Sieboldi, Max. Buna.

Taf. VIU, Fig. Oj,
Knospen spindelig. Schuppen hellbraun bis dunkelgrau-
braun, die Spitzen derselben sehr fein behaart, weisslich.
Zweige hin- u. hergebogen, die jiingeren graubraun griinlich,
die alteren weissgrau. Der Stamm hat eine grauweisse Far-

bung. Lent. deutlich. Mark von unregelmdssiger Form.

Fagus japonica, Max Inu-bara. Taf. VIII, Fig. 75.

Knospen sehr lang gestreckt bis 3 cm. lang, spindelformig,
zumeist lang gestielt. Schuppen von derselben Farbe wie bei
F. sylv. var. Sieboldi, aber ein wenig grau. Zweige schlank
hin- u. hergebogen, glatt, in der Jugend hellrothbraun, spater
grau. Kurztriebe eng eingeschniirt. Mark unregelmassig
eckig.

SHIRASAWA : LAUBHOLZER 1M WINTERZUSTANDE. 265

#.—Knospen kegelférmig.
Ilex macropoda, Mig. Aohada. Taf. IX, Fig. z, 2.

Knospen kegelig, kurz an der Triebe dicht stehend, locker
beschuppt. Knospenschuppen u. einjahrige Zweige hellbraun,
die alteren glanzend dunkelgrau Kurztriebe eng eingeschniirt.
Lent. deutlich. Mark rund.

Ginkgo biloba, L. Icho. Taf. IX, Fig. 3, 4.

Knospen kegelformig, kurz, Schuppen braun. Die jiingen
Zweige hell- od. graubraun glanzend, die alteren weisslichgrau.
Blattnarbe halbmondférmig. Lent. undeutlich. Mark un-
regelmassig. Zweige mit langen, eng _ eingeschniirten
Kurztriebe.

7.—Knospen spitz, walzenformig.
Ostrya virginica, Willd. Asada. Taf. IX, Fig. 75, 76.
Knospen walzenformig, zugespitzt od. eiformig. Knospen-
schuppen braun, od. dunkelréthlichbraun, teilweise gelblich-
griin, glanzend. Zweige dunkelrothlichbraun od. hellbraun
mit deutlichen Lenticellen. Blattnarbe halbmond- od. sichel-
formig. Mark eng und von unregelmassiger Form.

B.—Schuppen abwechselnd stehend.
a.—Schuppen auf zwei Seiten angeordnet.
*.—Knospen behaart.
Aphananthe aspera, Planch. Mukunoki. Taf. VIII, Fig. ro.

Knospen spindelig, zugespitzt etwas flach, gekriimmt, mit
kleinen Nebenknospen an der Trieben gedriickt, locker be-
schuppt, dunkelbraun grauweiss behaart. Zweige schlank,
besenformig, dunkelgrau, in der Jugend grauweiss, dicht
behaart. Mark rund.

**,—Knospen gar nicht od. wenig behaart.

Ulmus campestris, Sm. var. vulgaris, Planch. Kobu-nire.
Tay, Ie. Fig. 55.6:
Knospen klein kugelig, etwas abgeflacht, Schuppen schwarz-
lichbraun. Die jungen Zweige graubraun od. braun behaart,
die alteren grau. Die mehrjibrigen Triebe zeichnen sich
durch tiefe Risse in der Epidermis aus.

266 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Ulmus parvifolia, Facg. Aki-nire.

Knospen meist zu zwei beisammen sitzend, eiférmig, zuge-
spitzt, etwas abgeflacht, dunkelbraun. Zahlreiche, diinne fast
gleich lange, braune Seitenzweigen entwickeln sich aus der
Triebe. Lenticellen deutlich. Mark klein.

Ulmus campestris, Si. var, laevis, Planch. WHaru-nire.
Taf. IX, Fig. 8, 9.
Knospen kegelig, flach gekriimmt, 3-5 mm. lang, grau od.
dunkelbraun, schwach behaart. Zweige graubraun, dicker als
bei anderen Ulmusarten; die jiingeren ein wenig behaart.
Blattnarbe ziemlich gross, halbmondformig. Mark rund, klein.

Ulmus montana, Sm. vay. laciniata, Tvautv. Atsushi.

Taf TX iene
Endknospe etwas grésser als Seitenknospen, die letzteren
sind von verschiedener Form, spindelférmig, zugespitzt glan-
zend schwarzbraun. Zweige hin- u. hergebogen, auf der
Lichtseite graubraun glanzend, sonst grau. Blattnarbe ziem-

lich gross, halbmondférmig. Mark klein, rund.
Die Rindenfaser aller zu dieser Gruppe gehorigen Holzarten

ist meist ziemlich stark, u. das Holz biegsam.

b.—Schuppen von vier Seiten abstehend.
Carpinus japonica, BJ. Kuma-shide. Taf. IX, Fig. zo.
Knospen spindelig, etwas kantig, schlank u. scharf zuge-
spitzt. Schuppen hell- od. dunkelbraun glanzend. Zweige

schlank, hin- u. hergebogen, glanzend dunkelbraun. Lent.
deutlich. Mark rund.

Carpinus jedoensis, Max. Inu-shide. Taf. IX, Fig. zz.

Die Spitze der Knospen etwas gebogen, spindelig, vier-
kantig, dicker als bei Carpinus japonica. Schuppen hell-
rothbraun glanzend. Die einjaihrigen Zweige filzig behaart,
griinlichgrau.

Carpinus laxiflora, BJ. Soro. Taf. IX, Fig. 73.
Knospen spindelig etwas dick u. kurz, vierkantig. Die

jungen Zweige kahl, od. graugriin behaart. Der Stamm
grauweiss.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 267

Carpinus cordata, BJ. Sawa-shiba. Taf. IX, Fig. r2.

Endknospe 12 mm. lang, grésser als Seitenknospen ; beide
grosser als bei anderen Carpinusarten, spindelig, vierkantig-
die Spitze behaart. Knospenschuppen hellbraun, dunkel ge-
randert. Die jiingen Zweige graubraun, die 4lteren grau.
Lent. deutlich. Mark eckig.

Zelkowa acuminata, Planch. Keyaki. Taf. IX, Fig. r¢.

Knospen zumeist zwei beisammen dicht stehend, kegelfor-
mig, kantig. Schuppen dunkel- od. schwarzbraun, am Rande
ein wenig behaart. Zweige schlank, hin- u. hergebogen, braun
od. dunkelbraun. Blattnarbe elliptisch. Lent. rund, deutlich.
Mark rund.

IV.—Knospen von zusammengedruckten Blattchen
gebildet (nackt).
Picrasma quassioides, Benn. Nigaki. Taf. IX, Fig. zo.

Zwei unausgebildete Blattchen umfassen die Knospe, flach,
halbmondférmig, od. quadratisch, gelblich dunkelbraun, filzig
behaart. Blattnarbe gross, halbmondformig od. rundlich.
Zweige dunkelbraun, graulich. Lent. zahlreich, sehr deutlich.
Mark etwas weit. Die Rinde von bitterem Geschmack.

Hamamelis japonica, Szeb. ef Zucc. Mansaku.
Taf: IX, Bg. 17, 18.
Zwei unausgebildete Blatttchen beisammen stehend, wovon
das eine lang gestreckt, scalpelfo6rmig, lang gestielt. Sie sind
dicht filzig behaart, auf der Lichtseite dunkelgrau, sonst
hellbraun. Lent. ziemlich gross, deutlich. Blattnarbe halb-
kreisformig od. abgerundet dreieckig.

268 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

TABELLE fil.

ce oe = 27
Knospen ‘‘ gegenstandig.

I.—Die Knospen sind in der Blattnarbe verborgen.

Philadelphus coronarius, L. var. Satumi, Max. Baikwa-utsug1.
laf. Xs Big ee:

Blattnarbe dreieckig weissgrau, etwas vorspringend, in
der Mitte eine kleine Vorwolbung zeigen, unter welcher die
Knospe verborgen ist. Die jungen Zweige gerade, braun
etwas glanzend pfeifenrohrartig, sieht ganz diirr aus, die
alteren braungrau. Lenticellen undeutlich. Mark weit, rund.

II.—Zweige auffallend dick, Endknospen sehr gross.
Aesculus turbinata, B/. Tochinoki. Taf. X, Fig. 3.

Endknospe r cm. Durchmesser, eifo6rmig, zugespitzt etwas
gebogen ; die Seitenknospen kugelig, dunkelbraun sehr harzig.
Blattnarbe gross hellbraun gefarbt. An der Grenze der
jungen- u. alteren Zweige ringformige Einschniirung. Lent.
deutlich. Mark weit, rund u. braun gefarbt.

ITI.— Die Zweige sind zumeist dornspitzig.
Punica Granatum, lL. Zakuro. Taf. X, Fig. 6, 7.

Knospen klein pyramidenformig, kurz, rothlichbraun.
Zweige graubraun sehr schlank. Die oberen Peridermschich-
ten losen sich in Fasern ab. Lenticellen deutlich. Mark
ziemlich weit, rund.

Rhamnus japonica, Max. var. genuina, Max. Kuro-umemodoki.

Taf. X, Fig. 4, 5.

Knospen an der Triebe angedriickt, pyramidenformig, ziem-

lich lang, etwas gebogen. Schuppen dunkelbraun, am Rande

ein wenig behaart. Blattnarbe hoch, seitwarts pfriemenartige

Blattchen hervortretend. Die dornspitzigen Zweige derb,

hellgrau od. braungrau, glanzend. Die alteren Zweige dunkel-

braun. Blattnarbe halbmondformig od. sichelformig. Die

Rinde hat einen eigentiimlichen Geruch. Knospen oft nicht
gegenstandig angeordnet.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 269

IV.—Knospen von unausgebildeten Blattchen, nicht
von eigentlichen Schuppen umgeben.

A.—Knospen gestielt.
a.—Zweige (zweizeilig angeordnet) grau gefarbt.
Callicarpa mollis, Sicb. et Zucc. Yabu-murasaki.
Taf. X, Fag. 8.
Knospen sehr lang gestielt, kleinwalzig; Endknospe
grosser als Seitenknospen, die letzteren schmal, gekriimmt.
Zweige schlank, griinlich grau. Blattnarbe rundlich. Lent.
undeutlich. Mark sehr weit, 6 kantig.
Callicarpa japonica, Thunb. Murasakishikibu. Taf. X, Fig. zo.
Knospen kurz gestielt, graubraun kleinwalzig. Zweige
schlank, graubraun. Mark weit, rundlich.
Callicarpa purpurea, Fuss. Ko-murasaki. Taf. X, Fig. 9.
Knospen zumeist kugelig, sehr kurz gestielt, (seltner lang
gestielt) kleinwalzig. Lent. wenig, deutlich. Mark weit,
4 kantig.

b.—Zweige braun gefarbt.
Viburunum furcatum, B/. Mushikari. Taf. X, Fig. zz, 72.
Zwei lang gestreckte, unausgebildete, braun behaarte
Blattchen umfassen die Knospe. Zweige gerade, lang, griin-
braun, glanzend. Blattnarbe abgerundet dreieckig, mit 3
Gefassbundelspuren. Lent. wenig. Mark ziemlich weit.

B.—Knospen dicht an den Trieben sitzend.
Clerodendron trichotomum, Thunb. Kusagi. Taf. X, Fig. 73.

Zwei unausgebildete Blattchen umfassen die Knospe. Die
letztere an der Triebe fast senkrecht stehend, braunviolett,
dicht behaart. Zweige ziemlich dick, auf der Lichtseite
dunkelgraubraun, sonst grau, glanzend. Blattnarbe gross.

Lent. fein, sehr deutlich. Mark sehr weit, gefarbt.
‘Cornus ignorata, Koch. Kumano-mizuki. Taf. X, Fig. 14, 75.
Endknospe (von zwei unausgebildeten Blattchen gebildet)
grosser als Seitenknospen, keilformig, lang gestreckt. Die
letzteren nadelformig, von der stehenbleibenden Blattbasis
umgeben, dunkelgrau behaart. Zweige auf der Lichtseite

270 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

rothlich, sonst gelblichgriin, 6 kantig. Lent. wenig. Mark
rundlich, od. 6 kantig.

Premna microphylla, Turcz. Hama-kusagi. Taf. X, Fig. 16.

Knospen kugelig etwas gespitzt, hellbraun. Seitenknospen
flach. Die einjaéhrigen Zweige gerade, lang, hellbraun.
Blattpolster vorspringend. Blattnarbe halbkreisformig, ge-
wolbt. Mark ziemlich weit, elliptisch. Durch einen beson-
deren Geruch ausgezeichnet.

Vitex Negundo, L. Ninjinboku. Taf. X, Fig. 77.

Knospen sind unter der Blattachsel verborgen, so dass man
die Zahl der Blattchen nicht wahrnehmen kann, graubraun-
filzig behaart. Die einjahrigen Zweige viereckig, graubraun,
gerade u. lang. Blattnarbe sichelformig. Lent. klein, deut-
lich. Mark 4 kantig.

Vitex trifolia, L. var. unifoliolata, Schauer. Hamabo.

Knospen klein, etwas entfernt oberhalb der Blattnarbe
stehend, grau behaart. Die jiingeren Theile der Zweige dicht
behaart wie mit Sammet bekleidet, graubraun, 4 bis 5 kantig.
Mark weit, eckig. Kriechender Strauch nicht aufrecht
stehend.

Evodia rutecarpa, Benth. et Hook. Goshiyu. Taf. X, Fig. 78, 79.

Knospen flach, viereckig, hellbraunfilzig behaart. Zweige
ziemlich dick, gerade, dunkelcarmin roth, der jiingere Theil
schwach behaart. Blattnarbe gross halbmondformig, oft
ziemlich breit, meist mit 3 Gefassbiindelspuren versehen.
Lent. ganz deutlich. Mark weit, eckig. Holz u. Rinde
gelblich.

V.—End- u. Seitenknospen so von zwei Schuppen
dicht umgefasst, dass scheinbar nur eine Knos-
penschuppe vorhanden ist.

A.—Knospen unbehaart, glanzend.
a.—Knospen gestielt.
Viburnum opulus, L. Kanboku. Taf. X, Fig. 20. .

Knospen dem Stengel angedriickt, auf der Aussenseite
kugelig gewolbt, zugespitzt. Schuppen griinlich od. rothlich-
braun. Endknospe fehlt meist. Seitenzweige lang, nicht

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 271

abstehend, mehr od. weniger kantig. Junge Triebe braun,
altere grau. Mark weit, eckig.

Acer rufinerve, Sich. et Zucc. Uri-kaede. Taf. X, Fig. 22.

ISndknospe grodsser als Seitenknospe, 4 kantig, zugespitzt.
Seitenknospen eiformig, lang gestreckt, etwas gekriimmt.
Schuppen dunkelrothlich, glanzend. Zweige gerade, lang,
gelblichgriin, auf der Lichtseite réthlich, die alteren griin, mit
viereckigen, weissen Flecken versehen. Blattnarbe schmal,
mit 3 Gefassbiindelspuren. Lenticellen wenig. Mark weit,
rundlich.

Acer cratzgifolium, Sieb. et Zucc. - Meuri-kaede.
VGjPG) Fig. 23% 22.
Knospen eiférmig, zugespitzt; die Seitenknospen an der
Triebe angedriickt, meist mit einem Schuppenrisse auf dem
Riicken, kurz gestielt, dunkelréthlich. Zweige auf der
Lichtseite gelblichgriin od. rothlich, schwarzlich bekleidet.
Mark rund.

b.—Knospen sitzend.

Staphylea bumalda, Sieb. et Zucc. Mitsuba-utsugi.

Taf. X, Big. 27.

Knospen keilformig, riickwarts etwas gewolbt. Am Ende

der Triebe stehen immer je zwei Knospen beisammen.

Schuppen schwarzbraun. Zweige im ersten Jahre glanzend

dunkelbraun, die dlteren graubraun, feinrissig. Blattnarbe

schmal, herzformig, od. dreieckig. Lent. lang, deutlich.
Mark weit, rund.

Acer palmatum, Thunb. Kaede. Taf. X, Fig. 25, 26.

Zwei od. drei Endknospen beisammensitzend, kegelformig,
kurz, dunkelréthlich gefarbt. Zweige gelblichgriin, auf der
Lichtseite rothlich. Die stehenbleibende Korkschichte in der
Blattnarbe umbhiillt die Knospenbasis. Lenticellen undeut-
lich. Mark etwas weit, rund.

Salix multinervis, fv. ef Sav. Kori-yanagi. Taf. III. Fig. 7.
Zahlreiche Knospen an der Triebe angedriickt, kegelig,
klein. Bliitenknospen eiférmig, zugespitzt, die Spitze etwas
gekriimmt, gestielt. Zweige u. Schuppen glanzend roth, der
altere Theil der Zweige hellbraun. Blattnarbe sichelférmig.

272 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Salix rubra, L. Saruko-yanagi. Taf. III, Fig. zo.

Laubknospen spindelig, schlank, angeplattet. Bliiten-
knospen spindelformig, lang; die Spitze gekriimmt. Die
. Knospe u. Zweige auf Lichtseite roth, sonst gelblich griin.

B.—Knospen behaart.
Phellodendron amurense, Rupyv. Kiwada. Taf. X, Fig. 28.

End- u. Seitenknospe gleich gross, in der Mitte der Blatt-
narbe sitzend, von zwei Schuppen dicht umhiillt, hiigelformig,
dunkelbraun filzig behaart. Zweige sehr dick, gerade, die
jiingeren hellgelblichbraurr od. grau. Blattnarbe fusssohlen-
formig. Mark gross, rund, etwas gefarbt. Die Rinde zeich-
net sich durch gelbe Farbung aus.

Acer cissifolium, Koch. Mitsude-kaede. Taf. X, Fig. 27.

Endknospe von zwei kleineren Seitenknospen umgeben.
Seitenknospe an den Zweigen abgeplattet, von der Gestalt
eines Kafers. Knospenschuppen u. junge Triebe griinlich,
teilweisc grauweiss, die alteren Triebe glanzend grau. Blatt-
narbe schmal. Mark etwas weit.

Cornus Kousa, Buerg. Yamaboshi. Taf. X, Fig. 30, 37.

Knospen nur am Ende der Triebe, kegelig. Zwei dunkel-
violettbraun filzig behaarte Schuppen dicht zusammentretend.
Bliitenknospen kugelig, zugespitzt, mit mehr als zwei
Schuppen versehen. Blattnarbe schmal, umfasst die Knospe
ringsum. Zweige gerade, lang, dunkelbraun, teilweise weiss-
lich, mit zahlreichen, deutlichen Lenticellen. Mark eng.

Cornus Officinalis, Seb. Sanshiyu.
Taf. X, Pig. 20; te Naja eet
Endknospe groésser als Seitenknospen, zugespitzt, kantig,
grau behaart. Die einjahrigen Zweige 4 kantig, rothbraun,
weisslich bereift, ein wenig behaart. Blattpolster etwas vor-
springend. Blattnarbe dreieckig. Mark rund.

VI.—Seitenknospen eiférmig oder spindelig, fast
senkrecht abstehend.
Lonicera Morrowii, A. Gray. Kinginboku. Taf. XI, Fig. 4.

Knospen eiférmig, etwas kantig, locker beschuppt. Schup-

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 273

pen filzig behaart, hellbraunlich. . Endknospe einzeln. Zweige
ziemlich diinn, hellgrau, kahl. Die oberen Peridermschichten
lésen sich in Fasern ab. Mark innen hohl, am Rande braun.
Lonicera glacilipes, Mig. Uguisukagura. Ta/. XI, Fig. 2, 3.
Endknospe grésser als Seitenknospen, eiformig zugespitzt,
etwas kantig. Knospenschuppen’ diinnhautig, grau od.
graubraun. Zweige schlank, auf der Lichtseite grau, auf der
Schattenseite hellbraun. Diese Art zeichnet sich aus durch
die stehendenbleibenden, braunen halbkreisformigen Blattchen,
welche die Triebe ringsum umfassen. Mark rund.

Lonicera coerulea, L. Kuromino-uguiskagura.

Endknospe wie bei der Lon. gracilipes, locker beschuppt.
Seitenknospen etwas flach, u. zwei tibereinander von Zweige
abstehend. Zweige schlank, glanzend gelblich rothbraun ; die
ausseren Korkschichten losen sich faserig ab. Blattbasis sehr
hoch vorspringend, schuppenartig. Mark weit, etwas kantig.

VII.—An den Seitenknospen nur wenige Schuppen
(2 od. 3) sichtbar.

A.—Seiten- (End) knospen kantig.
a.—Knospen od. Zweige behaart, od. kleinwalzig.
Viburnum dilatatum, Thunb. Gamazumi. Taf. XJ, Fig. 5.

Endknospe vierkantig, zugespitzt. Seitenknospen flach, an
der Triebe angedriickt, filzig behaart, rothbraun. Zweige auf
der Lichtseite graubraun, kleinwalzig. Blattnarbe schmal.
Mark weit, rund.

Viburnum Sieboldi, Mig. Gomaki. Taf. XI, Fig. 7.

Knospen vierkantig, zugespitzt, langer gestreckt als bei der
vorigen Art, rothbraun schivach behaart. Zweige dunkelgrau
behaart. Blattnarbe gross, von der Gestalt einer fliegenden
Fledermaus. Mark rund. Die Rinde hat einen specifischen
unangenehmen Geruch.

Viburnum tomentosum, Thunb. Yabu-demari. Taf. XI, Fig. 6.
Knospen walzenformig, lang gestreckt, zugespitzt, etwas
kantig. Seitenknospen etwas gekriimmt. Schuppen u.

junge Zweige kleinwalzig, nicht behaart. Zweige graubraun,
Blattnarbe schmal. Mark rund.

274 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Viburnum Wrightii, Mig. Miyamagamazumi.
Taf. XI, Ftg. 8, 9.

Endknospe groésser als Seitenknospe, vierkantig zugespitzt.
Zwei lockere Schuppen an der Basis der Knospe. Die
Bliitenknospe am Triebende vergekehrt kegelférmig. Schup-
pen dunkelrothlich, kleinwalzig. Zweige gerade, grauroth-
braun. Blattnarbe schmal. Lent. deutlich, gering.

b.—Knospen nicht od. wenig behaart.

Fraxinus Sieboldiana, B/. Shioji. Taf. XI, Fig. 72.

Zwei kleinere Knospen umgeben die Endknospe, die letztere
wesentlich grosser als die Seitenknospen, vierkantig, kurz.
Schuppen derb, gekielt, dunkelbrau, an der Basis weiss mehlig.
Blattnarbe schmal, umfasst fast die ganze Knospe. Zweige
sparrig, hellgraugrtin. Lent. ganz deutlich, lang. Mark weit,
rund.

Fraxinus Bungeana, DC. var. pubinervis, B/. Toneriko.
laf. XI, Pigenro, ge
Knospen ahnlich wie bei der Fr. Sieboldiana; aussere zwei
Schuppen in der Mitte der Knospe meist frei zusammentretend.
Die jungen Zweige etwas kantig (4), gerade, an der Basis
gewolbt, dunkelbraun, gelb, teilwcise weiss gefleckt. Lent.
undeutlich. Mark undeutlich viereckig.

Fraxinus longicuspis, Sieb. cf Zucc. Kobano-toneriko.
laf. XI, Fig, 23s
Endknospen etwas groésser als die Seitenknospen, weniger
scharf kantig, etwas flach. Seitenknospen fast senkrecht
abstehend. Schuppen schwarlich bereift. Zweige grau, rund-
lich ; Blattpolster etwas gewolbt. Blattnarbe halbmondformig.
Lent. fein, zahlreich. Mark eng, eckig.

Chionanthus retusa, Lindl. et Paxt. Hitotsubatago.
Taf. XI, Figs ag.
Endknospe grésser als Seitenknospe schaf 4 kantig, kurz.
Schuppen dunkelbraun Zweige diinner als die der vorigen
Arten, fast senkrecht abstehend, braungrau. Blattpolster
hoch. Blattnarbe halbelliptisch. Lent. deutlich, rund. Mark

eng.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 275

B.—Seitenknospen angedriickt, an der dem Zweige zuge-
wendeten Seite abgeplattet.
a.—Knospen u. Zweige nicht behaart.
a.—Blattnarbe schmal.

Acer argutum, Maxim. Asanoha-kaede. Taf. XI, Fig. 77, 78.

Knospenam Triebende gehauft, die Mittelern lang gestreckt,
von zwei Schuppen dicht umbhiillt, grésser als die tibrigen.
Seitenknospen von der Gestalt eines Kafers. Schuppen u.
einjahrige Zweige dunkelrothlich, dicht bereift, altere Zweige
gelblich, auf der Lichtseite rothlich. Zwei schmale Blattnar-
ben zusammenstossend. Lent. wenig. Mark rund.

Acer distylum, Szeb. ef Zucc. Hitotsuba-kaede.
ITE DOG 1B ee Oe
Eine grossere Endknospe mit zwei kleineren Seitenknospen
seitwarts, kafergestaltig. Unausgebildete Blattchen umgeben
haufig die Knospe. Knospenschuppen u. Zweige dunkelroth,
graubraunfilzig behaart. Blattnarbe schmal, je zwei zusam-
menstossend. Mark rund.

Cercidiphyllum japonicum, Szeb. ct Zucc. Katsura.
Taf xh, Fig. 10.
Am Ende der Triebe zwei Knospen beisammen abstehend,
End- u. Seitenknospen gleich gross, kegelig. Eine an dem
Zweige abgeplatte Schuppe umhiillt die dussere der Knospe,
u. eine andere die innere derselben; die erstere ist dunkel-
rothbraun, die letztere rothlich. Zweige schlank u. lang, an
der Lichtseite rothbraun, sonst hellbraun. Blattpolster vor-
gewolbt. Blattnarbe schmal. Lent. fein, deutlich. Mark
eng.

#.—Blattnarbe gross rundlich od. halbmondformig.

*. —Baumgewachse.

Euseaphis japonica, Pax. Gonzui. Taf. XI, Fig. 20.

Am Ende der Triebe zwei Knospen; sie sind keilformig,
seitwarts gewolbt. Schuppen schwarzlich roth. Zweige ziem-
lich dick, auf der Lichtseite glanzend dunkelroth, sonst griin-
lich gelbroth. Lent. wenig. Mark weit, rund.

276 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

Acer japonicum, Thunb. WHauchiwa-kaede. Taf. XI, Fig. 2z.
Endknospe zwei beisammenabstehend, eiférmig, zugespitzt,
3 od. 4 Schuppen sichtbar, schwarzlichroth gefarbt. Die
stehenbleibende Korkschichte der Blattbasis bildet die Blatt-
narbe, sie ist gross u. unterseits eckig, am Rande behaart.
Mark weit, eckig.
Acer Sieboldianum, Max. Kohauchiwa-kaede.
Knospen sind kiirzer, u. Zweige schlanker als bei Acer
japonicum.

Hydrangea hortensis, Smith. var. japonica, Maxim. Gaku.
Taf. XI, Fig. 23:
Endknospe grésser als Seitenknospe, spindelformig 4 kantig,
die dusseren zwei Schuppen leicht ablésbar. An der Seiten-
knospe 3 Schuppen sichtbar, dunkelrothbraun. Zweige glan-
zend hellbraun. Blattnarbe gross, abgerundet dreieckig od.
halbmondformig. Mark sehr weit.

Hydrangea Thunbersgii, Sie). Amacha.
Knospen u. Zweige sehr ahnlich wie bei der vorgenannten
Mites
Hydrangea hortensia, DC. Ajisai. Taf. XI, Fig. 24.
Endknospe wesentlich grésser als Seitenknospe, eiformig,
zugespitzt. Die ausseren zwei Schuppen leicht ablosbar, die
inneren unausgebildeten Blattchen bleiben schallen, glanzend
gelblichgriin. Zweige dick, hell- od. graubraun. Blattnarbe
gross, halbmondformig od. etwas eckig. Mark sehr weit.

b.—Knospen u. Zweige behaart.
Ligustrum Ibota, Sieb. Ibotanoki. Taf. XI, Fig. 25.
Seitenknospen an der Zweige abgeplattet, keilformig, dunkel-
graufilzig behaart. Zweige schlank, filzig behaart, schmutzig
bestaubt. Blattpolster vorspringend; Blattnarbe halbmond-
formig. Lenticellen undeutlich. Mark eng.

VIII.—An den Seitenknospen mehr als 3 Schuppen
sichtbar.
A.—Knospen u. Blattnarbe gross.
a.—Knospen kugelig.

SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE. 277

Sambucus rasemosa, L. var. Sieboldiana, Mig. Niwatoko.
Knospen kugelig, gestielt, od. verkehrt eiformig, roth bis
violettgefarbt. Die zahlreichen Schuppen umgeben die Knospe
ziemlich locker. Oefter stehen noch 1-3 Knospen unter der
Hauptknospe. Zweige bogenformig gekriimmt, grau, mit
deutlichen Lenticellen versehen. Mark sehr weit, braun

gefarbt.

b.—Knospen kantig.
Syringa vulgaris, 2. Murasaki-hashidoi. Taf. XJ, Fig. 28.
Knospen 4 kantig, zugespitzt, kurz. Schuppen carminroth,
glanzend. Zweige dunkelgrau, schmutzig bestaubt. Blatt-
narbe ziemlich gross, dreieckig, od. halbmondformig. Lent.
ziemlich deutlich. Mark eng, eckig.
Syringa japonica, Max. MHashidoi. Taf. XI, Fig. 27.
Knospen 4 kantig, scharf gespitzt, Schuppen hellbraun.
Zweige dunkelegrau, Blattnarbe gross, halbmondformig. Blatt-
polster vorspringend. Lent. gross, deutlich. Mark eng, eckig.

B.—Knospen weniger gross, von der Schuppe sehr lose
umhiillt od. nicht ; Blattnarbe ziemlich gross.
a.—Knospen kegel- od. spindelformig.
a.—Knospen unbehaart.
Enonymus oxyphyllus, Mig. Tsuribana. Taf. XII, Fig. rz.
Knospen spindelférmig schlank, sehr lang, dicht beschuppt,
auf der Lichtseite dunkelrothlich, sonst dunkelgelblich griin.
Zweige diinn auf der Lichtseite rothbraun, auf der Schatten-
seite griin, die alteren braun. Blattnarbe dreieckig od. halb-
mondférmig.’ Lent. sehr wenig. Mark eckig.
Forsythia suspensa, Vahl. Rengyo. Taf. XII, Fig. 3.
Bliitenknospen spindelig, locker beschuppt. Zweige u.
Schuppen gelblichgrau. Blattpolster vorspringend. Blattt-
narbe halbmondformig, in der Mitte mit einer hervortretenden
Gefassbiindelspur. Mark hohl.
Viburnum phlebotrichum, Sicb. et Zucc. Otokoydzome.
Taj Xl, Fig. 2.
Endknospen (Bliiten) kugelig, zugespitzt. Seitenknospen
liber der Blattnarbe aufrecht stehend, spindelig, etwas flach ;

278 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

4-5 Schuppen sichtbar. Schuppen schwarzlich roth, glan-
zend, Zweige weisslichgrau. Lenticellen wenig. Mark rund.

§.—Knospen behaart.
Acer nikoense, Maxim. Megusurinoki. Taf. XII, Fig. 4.

Knospen spindelig, lang, an der Spitze der Langtrieben
gehauft, 4-5. Schuppen schwarzlichbraun, graubraunfilzig
behaart. Blattnarbe schmal, umfasst die Knospe, am Rande
mit langen Haaren. Zweige schwarzlichbraun schmutzig.
Lenticellen zahlreich, deutlich. Mark weit.

b.—Knospen kantig.
a.—Knospen von eigentlichen schuppen umgeben.

Acer carpinifolium, Sieb. ef Zucc. Chidorinoki.
Taf. XII, Fre. &2.0:
Am Ende der Triebe 2 od. 3 Knospen beisammenstehend,
scharfkantig dunkelroth, an der Basis schwach _behaart.
Zweige rothlichbraun, auf der Schattenseite graubraun.
Blattnarbe umfasst die Knospe. Lenticellen gross, sehr deut-
lich. Der Stamm grauweiss.

Evonymus alatus, Fr. ef Sav. Nishikigi. Taf. XII, Fig. 8.

Drei Knospen an der Spitze der Triebe, kantig, zugespitzt.
Schuppen dunkelrothbraun weisslich gefleckt. Zweige griin,
auf der Lichtseite rothbraun, mit 2 od. 4 herablaufenden
Korkleisten. Mark 4 kantig, die Kante der Lage der Kork-
leisten entsprechend.

Evonymus europeus, L. var. Hamilitonianus, Wax. Mayumi.

Taf. XII, Fug. 9.

Knospen ein wenig scharfkantig, etwas flach, von Schuppen

lose umhiillt. Schuppen dunkelrothlichbraun, weisslich

gerandert. Zweige griinlichroth. Blattpolster vorspringend.

Blattnarbe ziemlich gross, halbmondformig. Mark rund.
Das Holz hat gelbe Farbung.

Calycanthus precox, L. Robai. Taf. XII, tg. 75.

Knospen eiformig etwas kantig. Schuppen braun. Zweige
braun, weisslich bekleidet, so haben sie eine hellbraue Farbung
glanzend. Blattnarbe ziemlich gross, herzformig. Lent.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 279

deutlich. Mark weit, 6 kantig. Im Winter kugelige Bliiten-
knospen tragend.

Deutzia scabra, Thunb. Utsugi. Taf. XII, Fig. zo.
Knospen 4 scharfkantig, zugespitzt, etwas lang. Schuppen
hellbraun, dunkler gerandert, graubraun mehlig. Zweige

braun od. dunkelbraun. Die obere Peridermschichte rissig.
Mark hohl.

Diervilla grandiflora, Sieb. et Zucc. Hakone-utsugi.
Daf. ALG hie. 13. Ld.
Endknospe grosser als Seitenknospen, die letzteren bogig_
gekriimmt, locker beschuppt. Schuppen graubraun; Zweige
von derselben Farbe. Blattnarbe sehr gross. Lent. gross,
deutlich. Mark sehr weit.

Diervilla japonica, Sieb. ef Zucc. Tani-utsugi.
Taf. XIf, Fig. 72.
Knospen locker beschuppt. Schuppen u. Zweige dun-
kelrothbraun, teilweise schwarzlich. Blattnarbe gross. Mark
weit.

Ligustrum medium, Fr. ef Sav. Oba-ibotanoki.
Lay. XL, Big. 20.
Knospen 4 kantig, zugespitzt, locker beschuppt. Schup-
pen dunkelcarminrothlich, am Rande hellbraun, von ver-
trocknetem Aussehen. Blattpolster sehr hoch vorspringend,
Blattnarbe halbmondfoérmig. Lent. wenig vorhanden, aber
deutlich. Mark rundlich od. schwach eckig.
Acer Ginnala, Maxim. Kara-kogi. Taf. XII, Fig. 7.
Endknospe etwas grosser als Seitenknospen, beide 4 kantig,
kurz. Schuppen braun, weisslich bereift. Zweige schmal,
Blattnarbe schmal, verkehrt V-formig. Lent. deutlich. Mark
rund.

&.—Knospen von pfriemenformigen Schuppen umgeben.

Deutzia gracilis, Sich. et Zucc. Hime-utsugi. Taf. XII, Fig. rz.

Knospen kugelig, flach, od. von der form einer gleichschen-

klichen Dreieckig, bogig gekriimmt. Schuppen u. Zweige

grau, od. graubraun dunkler gefleckt. Blattnarbe schmal
sichelformig. Mark hohl.
c.—Knospen kugelig.

380 SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE.

Catalpa Kempferi, Sieh. et Zucc. Kisasage. Taf. XII, Fig. 27.
Knospen zumeist quirlstandig (3), kugelig, von dunkel-
braunen Schuppen sehr locker umgeben, vom Aussehen
gedffeneter Kieférnzapfen. Zweige dick graubraun, die
alteren dunkler. Blattnarbe gross, rundlich, gewolbt. Lent.
deutlich. Mark sehr weit von unregelmassiger Form.

Hydrangea paniculata, Sieb. Norinoki. Taf. XJ, Fig. 22.

Endknospe grodsser als Seitenknospen, kugelig, am Trieb-
ende ganz dicht sitzend, locker beschuppt, dunkel od.
schwarzlichbraun. Die Seitenknospen kegelformig, kurz onv
der Triebe fast senkrecht abstehend. Zweige ziemlich dick,
gerade pfeifenrohrartig, dunkelrothlich braun. Blattnarbe
gross. Lent. deutlich. Mark sehr weit. Knospen oft quirl-
standig (3).

C.—Knospen an der Spitze der Triebe gross, an der Basis
klein.

a.—Blattnarbe klein.
Acer pyenanthum, C. Koch. WHana-kaede. Taf. XII, Fig. 709.
Endknospe wesentlich grdsser als Seitenknospen scharf-
kantig, zugespitzt. Seitenknospen etwas flach. Schuppen
dunkelréthlich, am Rande wenig behaart. Die einjahrigen
Zweige, glanzend roth. Biattnarbe schmal. Lent. wenig, ~
deutlich. Mark weit, rund.

b.—Blattnarbe ziemlich gross.

Acer purpurascens, Fr. et Sav. Kaji-kaede. Taf. XII, Fig. 20.
Knospen kantig, zugespitzt, Schuppen dunkelbraun, grau-
weiss, dicht filzig behaart. Zweige etwas dick, graubraun
glanzend, fein rissig. Blattnarbe von nebenstehender Form
&, Lenticellen vorspringend. Mark rundlich, od. von un-
regelmassiger Form.
Acer pictum, Thunb. Itaya-kaede. Taf. XII, Fig. 78.
Endknospe eiformig, etwas kantig, locker beschuppt. Sei-
tenknospen an den Trieben abgeplattet, riickwarts gewolbt,
kafergestaltig. Schuppen dunkelrothlich, glanzend. Junge
Zweige grauweiss od. graubraun glanzend wie lackiert.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 281

Lent. deutlich. Blattnarbe schmal, umfasst die Knospe. Die
Rinde milchsaftig.

Acer pictum, Thunb. var. P
Die Form u. Farbe der Knospe ahnlich wie bei der Acer
pictum. Junge Zweige rothbraun, glanzend, rissig, die alteren
schwarzlichbraun. Blattnarbe schmal. Lent. deutlich. Mark
welt.

D.—Knospen klein.
a.—Schuppen gegenstandig angeordnet.
a.—Zweige diinn.
Abelia serrata, Sicb. et Zucc. Kotsubane-utsugi.
Taf. XII, Fig. 27.
Knospen sehr klein, 2-4 mm. lang, eiférmig zugespitzt,
etwas kantig. Schuppen braun od. rothbraun, am Rande
ein wenig behaart. Zweige auf der Lichtseite dunkelgrau, auf
der Schattenseite hell- od. dunkelbraun, glanzend, dltere grau.
Lent. undeutlich. Mark weit.
8.—Zweige auffallend dick.
Paulownia tomentosa, Baill. Kiri. Taf. XII, Pig. 76.
Knospen klein, dicht sitzend, Schuppen dunkelgraubraun.
Zweige dick, dunkelgriinlichgrau, mit hellen Flecken u. zahl-

reichen rauhen Lenticellen versehen. Blattnarbe gross, rund-
lich. Mark gefachert.

b.—Schuppen spiralig angeordnet.

IX.—Zweige kletternd (Schlinggewachse).

1.—Knospen sind von einer od. zwei Schuppen umgeben.
Pedelia foelida, L. WHekuso-kazura. Taf. XII, Fig. 22.
Knospen klein, kegelformig, kurz. Schuppen u. Zweige
hellbraun ein wenig behaart. Blattnarbe etwas gross, halbel-
liptisch gewolbt. Mark bandférmig. Die Rinde hat einen
unangenehmen Geruch. Im Winter bleiben die glanzend
hellbraune, kleinen kugeligen Friichte.
2.—Knospen vou mehreren Schuppen umgeben.
A.—Zweige mit Ranken.

Clematis japonica, Thunb. Tsurigane-kazura. Taf. XII, Fig. 23.

282 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

Knospen kugelig an der Triebe senkrecht stehend. Schup-
pen filzig behaart, grauweiss, glanzend, aber die dussersten
zwei Schuppen sind derb, schwarzbraun. Zweige dunkel-
braun, glatt; die jiingere ein wenig behaart. Lent. undeut-
lich. Mark eng.

B.—Zweige ohne Ranken.
Schizophragma hydrangeoides, Sieb. ef Zucc. Iwagarami.
Taf. XU, Pigwe7
‘Knospen cylindrisch, locker beschuppt, braun. Zweige
ziemlich dick; die jiingeren hellbraun behaart; die alteren
grau. Blattnarbe gross, dreieckig, od. halbmondformig. Mark
gross etwas eckig.

Hydrangea petiolaris, Sieb. ct Zucc. var. cordifolia, Max.
Gotozuru. Taf. XII, Fig. 25.
Endknospen grosser als Seitenknospen, lang gestreckt,
von Pyramidenform, locker beschuppt, hellgelb od. hellroth.
Zweige braun, lang, mit zahlreichen Luftwurzeln versehen.
Blattnarbe schmal. Lent. undeutlich. Mark weit, griinlich
gefarbt.

SHIRASAWA: LAUBHOLZER IM WINTERZUSTANDE. 283

TABELLE IV.
Zweige ‘ quilstandig. ”

I.—Knospen sind von unausgebildeten Blattchen
umgeben.
Clethra barbinervis, Sicb. et Zucc. Ryobu. Taf. XIII, Fig. z, 2.
Drei unausgebildete Blattchen umgeben die Knospe, drei-
kantig, die Spitze gekriimmt, weissbraun dichtfilzig behaart.
Zweige sparrig, gerade, lang, hellbraun, die jungeren Theile
grauweiss, mehlartig. Die zahlreichen dreieckigen Blattnar-
ben in der Nahe der Knospe zusammenstossend. Mark sehr
weit, rundlich.

II.—Endknospe sehr gross, einjahrige Zweige auffal-
lend dick.

Sterculia platanifolia, L. Aogiri. Taf. XIII, Fig. 4g.
Endknospen viel grésser als Seitenknospen, kugelig, dunkel-
braun filzig behaart. Zweige dick, griin, teilweise schwarz-
lich. Blattnarb gross. Lent. deutlich. Mark rund.
Idesia polycarpa, Max. ligiri. Taf. XIII, Fig. 3.
Zahlreiche, saftige Schuppen umgeben die Endknospe, sie
sind dunkelrothbraun, kahl, harzig. Zweige dunkelrothbraun,

die altere grau. Blattnarbe rundlich. Lenticellen deutlich.
Mark weit.

Aleurites cordat, DC. Abura-giri. Taf. XIII, Fig. 5.
Endknospe_ kugelig, locker beschuppt, dunkelréthlich.

Zweige dunkelroth griinlich. Blattnarbe gross, braun gefarbt.
Lent. deutlich, lang. Mark weit, rundlich.

III,—Knospen sitzend, von mehreren Schuppen um-
geben, an der Langtrieben einzeln od. gehauft.
A.—An der Langtriebe einzeln.
a.—Knospen kugelig- od. eiformig.
Cornus macrophylla, Wall. Mizuki. Taf. XIII, Fig. 6, 7.

Knospen eiformig, lang, locker beschuppt, glanzend dunkel-

284 SHIRASAWA : LAUBHOLZER IM WINTERZUSTANDE.

rothbraun, kahl. Zweige sparrig, lang, vogelfussf6rmig, dun-
kelroth glanzend. Blattnarbe halbkreisformig. Mark rundlich.
Andromeda cernua, Mig. Yoraku-tsutsuji.

Taf. XIII, Fig. 8, 9.

{Knospen eiformig. Die ausseren Knospenschuppen leicht

ablosbar, braun, die inneren roth gefarbt. Zweige diinn,

gerade, die ein-jahrigen braun, die alteren graubraun. Blatt-
narbe klein. Mark weit.

b.—Knospen spindelig.
a.—Schuppen behaart.
Rhododendron dilatatum, Mig. Mitsuba-tsutsuji.

Taf. XIII, Fig. ro, 77.
Knospen spindelig, die Spitze etwas gekriimmt. Schuppen
hellbraun behaart. Zweige gerade, die jiingeren hellbraun, die
alteren dunkelgrau. Blattnarbe halbmondformig. Mark rund,

weit.

8.—Schuppen unbehaart.
Rhodopendron sinenses, Sw. Kirengetsutsuji.
Taf. XU, Fae es
Knospen sehr gross, spindelig, kurz. Schuppen rothbraun,
teilweise schwarzlich, am Rande grauweiss, wenig behaart.
Junge Zweige hellbraun glatt, altere grau. Blattnarbe fuss-
sohlenformig. Mark eng, von unregelmassiger Form.
Enkyanthus japonicus, Hook. Dodan. Taf. XIII, Fig. 73, 14.
Knospen spindelig, kurz. Schuppen spiralig angeordnet,
carminrothlich, sehr schén. Zweige diinn, gerade, die jiinge-
ren rothbraun, od. hellbraun, die alteren grau od. graubraun.
Blattnarbe dreieckig.
B.—An der Langtrieben gehauft.
Rhododendron Schlippenbachii, Max. Kurofune-tsutsuji.
Taf. XIII, Fig. 75.
Knospen gleich gross, od. seitwarts der grossen Knospe
zahlreiche kleine flache Knospen zusammensitzend, hellbraun
dichtfilzig behaart. Zweige ziemlich dick, gerade, lang, grau-
braun, sehr rauh. Blattnarbe gross. Mark von unregelmas-

siger Form.

UBERSICHT UND REGISTER.

TABELLE I.

Die Knospen sind an der Langtriebe spiralig
angeordnet.

I.—Die Knospen sind in der Blattnarbe verborgen.

Taf. I, Fig. z Robinia Pseudacacia, L—aYmUyva .. .. .. «. «. 230

II.—Knospen gestielt.
a.—Eine grossere Schuppe umhiillt fast die ganze
Knospe.
Taf. I, Fig. 2. Alnus incana, Willd. var. glauca, Ait— ¥ VAY) .. .. 230
Meas He Pho Go INVES yey, Sulos Gr ZONA ES 5a no 55 600 oo ILE

Taf. I, Fig. g. Alnus firma, Sieb. ef Zucc—vvyY TY .. 2. «so ss we 23
Taf. I, Fig. 5. Alnus viridis, D.C. var. sibirica, Regel— = ¥ YY ) * S60 Beh

b.—Mehrere Schuppen umgeben die Knospe spiralig.
Taf. I, Fig. 6. Ribes fasciculatum, Steb. et Zucc.—V¥ FYYVYY .. .. «+ 231

III.—Knospen von unausgebildeten Blattchen gebildet.
A.—Zweige auffallend dick.

a.—Mark gefachert.

Taf. I, Fig. 7. Juglans Sieboldiana, Maz.—F=A7PN=2 2. «1 oe oe «- 231
Taf. I, Hig. 8 Juglans cordiformis, Maxi—eE xX Fyr=s .0 «1 22 oe os 232
Taf. I, Fig. ro. Pterocarya rhoifolia, Sieb. ef Zucc.— UNF 1. we os 232
Taf. I, Fig. 9. Juglans regia, L. var. sinensis, Cas—FYF7NX= «.. «. 232

8.—Mark nicht gefachert.

Taf. II, Fig. zo. Cedrela chinensis, $uss.—¥¥ vy Fv GO 0G 00 00 6m YBw
Taf. I, Fig. 72. Rhus vernicifera, Mig.—Jrmy .. 1. «2 os oe oe oe 232
Taf. I, Fig. ra. Rhus trichocarpa, D.C.—¥ YY... 2. oe we oe oe 233
Nhe We Jirte, Ta, AAG) RUSECCENIEEY, IS OG 8G 0D 00 oo OO 40) oO 25%}
Taf. I, Fig. 13. Rhus sylvestris, Sieb. ef Zucc.—¥ VAVwe «2 oe 2s oe 233

B.—Zweige weniger dick.
a.—Zweige ohne Stacheln od. Dornen.
Taf. I, Fig. 16. Walesia hispida, Benth. et Hook.—% 7 URF og. 60) OF Ie

286 SHIRASAWA: UEBERSICHT UND REGISTER.

Taf. I, Fig. 77. Halesia corymbosa, Benth. ef Hook—TF¥ffF .. «- «
Taf. I, Fig. 19. Rhamnus crenata, Sieb ef Zuce—4 Y PR «- oo ce ve
Taf. I, Fig. 20. Meliosma myriantha, Sieb. et Zucc—TA°T*X ..
Taf. I, Fig. 28. Mallotus japonica, Muell. Ary —FRHAHY. ..

b.—Zweige mit Stacheln od. Dornen.
a.—Zweige mit Stacheln.
Taf. II. Fig. 34. Czsalpinia sepiaria, Roxb—2>¥ FY ASF .. 22 ewe
b.—Zweige mit Dornen.
Taf. II, Fig. 2. Eleagnus umbellata, Thunb—7% Fz .. .. .. oe
Taf. II, Fiz. z. Eleagnus longipes, A. Gray.—}'‘Y Fz .. «2 2 os «

IV.—Knospen sitzend, so klein, dass man die Stellung
u. Zahl der Schuppen nicht mehr deutlich wahrneh-
men kann.

a.—Zweige mit metamorphosierten Seitenzweigen.
Lycium chinense, Will.—9 a) =. ac 220 See ce) eee
&.—Zweige mit Dornen.

fof. Tl, Fig..5. ‘Zizyphus volearis, Lam.— 99/1 X) 0) ee) ee eae
Taf. II, Fig. 6. Paliurus aubletia, Rem. et Sch—”AvF+YR.. «-

y-—Zweige ohne metamorphosierte Nebenzweige
od. Dornen.

Taf. II, Fig. 8. Ulex Sieboldi, Mig—JA2ERX-. 2. 2. 2s 2e oe oe
Taf. II, Fig. 7. Vex geniculata, Max.—JYWYVYTIXERE .. 2. oe oe
Hibiscus syriacus, b.—-2» 73" S.J sm a) en ees

V.—Knospen sitzend, Zahl der Schuppen od. zusammen-
gedriickten Blattchen undeutlich.

A.—Einjahrige Zweige auffallend dick.
a.—Zweige ohne Stacheln od. Dornen.
a.—Knospen filzig behaart.

234

234

- 235

235

235
235

235
236
236

Ailanthus glandulosa, Desi—YyYa .. «2 «2 «2 + 236

Taf. II, Fig. 9. Melia japonica, G. Don.—_tk vey .. ss tes. tele we e230

Taf. II, Fig. 12. Rhus semi-alata, Murr. var. Osbeckii, D.C.—X nF .- «- 236
?.—Knospen nicht od. wenig behaart.

Taf. II, Fig. 13. Ehretia acuminata, R. Br.—F>¥ J) ¥+. oe oe oe e+ 237

Taf. II, Fig. rz. Sapindus Mukurosi, Gaertn—B7 HY .. «1 oe

. 237

SHIRASAWA: UEBERSICHT UND REGISTER. 287

b.—Zweige mit Stacheln od. Dornen.
SEITE
Taf. III, Fig. 3. Zanthoxylum ailantoides, Sieb. ef Zucc—HFA)YYEY 237
B.—Einjahrige Zweige weniger dick.
a.—Zweige ohne Stacheln od. Dornen.
a.—Knospen nicht od etwas behaart.

Taf. TI, Fig. 16. Albizzia Julibrissin, Boivu—AB .. .. 1. 1. we os 237

£.—Knospen behaart.
Taf. II, Fig. 17. Sophora japonica, L—mwyya ob co 6p pe Go ARS
Taf. II, Fig. 20. Marlea platanifolia, Sieb. ef Zucc.—y y ) & 55 60. ao. HEYe
Taf. II, Fig. 22. Styrax Obassia, Sieb. et Zucc—A7YYRFF .. .. «. 238
Taf. II, Fig. 2r. Styrax japonicum, Sieb. et Zucc.—xca")% .. .. .. «. 238

b.—Zweige mit Stacheln od. Dornen.
a.—Zweige mit Dornen.

Taf. II, Fig. 78, 19. Cudrania triloba, Hance.—r~vy Fr . co 60 66 OO Hef
Taf. II, Fig. 14, 75. Gleditschia japonica, Mig.—4 {HF .. .. 1. .. 239

b.—Zweige mit Stacheln.

Taf. II, Fig. 23. Zanthoxylum piperitum, D. C.—#yt4 .. 1. 1. 4. 239
Taf. III, Fig. 2. Zanthoxylum alatum, Roxb.—JayYyesy .. .. 2. 2. 239
Taf. III, Fig. z. Zanthoxylum schinifolium, Sieb. et Zucce.— 4 X4Fy t Wi se 239

VI.—Knospen sitzend, mit einer bezw. zwei Schuppen.
A.—Knospen ausgesprochen kegelférmig, von der
Blattnarbe ringformig umgeben.

a.—Knospen od. junge Zweige behaart.
Taf. III, Fig. 5. Salix brachystachys, Benth—+ h¢nay Dae eee

+ 239

b.—Knospen od. Zweige nicht behaart.
Taf. III, Fig. 6. Salix gracilistyla, Mig—7 n ¥ +¥.. Ramer
Taf. III, Fig. 8, 9. Salix japonica, Thunb.—y-s~¥ +¥ .. . 240

Taf. III, Fig. 4. Ficus carica, L.—4 F3°7.. .. .. oS O06 AO oo A

B.—Knospen nicht kegelformig, stehen iiber der
Blattnarbe. Nur eine beiderseits kantige Schuppe
(aus zwei Schuppen verwachsen).

a.—Zweige dottergelb.

Taf. Il, Fig. rz. Salix purpurea, L.— Hav¥ + ¥
DEMS Chitin So od) oor isc eee

«= 240
66 00 oo Bi

288 SHIRASAWA: UEBERSICHT UND REGISTER.

b.—Einjahrige Zweige roth, od. braun glanzend wie

lackiert.
a.—Knospen dichtfilzig behaart.

Taf. III, Fig. 178, 19. Hovenia dulcis, Thunb—F vy K+ vy
Taf. III, Fig. 75. Magnolia Kobus, D.C.—a 7 vy
Taf. III, Fig. 14. Magnolia obovata, Thunb—€ 7vvy

#.—Knospen nicht behaart.
Taf. III, Fig. 16, 17. Andromeda ovalifolia, Wall—yyv7y:

Taf. III, Fig. 2z. Stachyurus precox, Sieb. et Zucc.—*% IF ..
Taf. III, Fig. 20. Itea japonica, Oliv.—xX 4 + A
Taf. III, Fig. 12. Salix Thunbergiana, Bl.—_Z43¥7*¥ ..

Taf. V, Fig, 7, 8. Helwingia ruscifolia, Willd.—.\+ {n°

C.—Zweige nicht auffallig gefarbt, Knospen lang ge-

streckt.
a.—Knospen nicht behaart, walzig.

Taf. III, Fig. 22. Magnolia hypoleuca, Sieb, ef Zucc.—m*7 ) =

we

f#.—Knospen sehr wenig od. gar nicht behaart,

zugespitzt.
Taf. VI, Fig. 4, 5. Lindera hypoleuca, Max.—7n#yY
7-—Knospen behaart, dem Stengel angedriickt.

Taf. III, Fig. 13. Salix viminalis, L—4 x¥ +¥

- 243

. 243

d.—Zweige griin, od. braun, Knospen keil-(umge-

kehrt)-formig.
a.—Blattnarbe rundlich.

Taf. III, Fig. 23. Diospyros Kaki, L. fl—H% .. «2 22 oe oe
Taj. III, Fig. 24. Diospyros Lotus, L.—v 4 7+

Taj. IV, Fig. 2. Broussonetia Kasinoki, Siebh.—A7Y a
Taf. IV, Fig. zr. Broussonetia Kasinoki, Sieb.var—EAATS ..
Taf. IV, Fig. 3. Broussonetia papyrifera, Vent—H) #.. .. «-

8.—Blattnarbe nicht rundlich.
*,—Zweige diinn.

Stillingia sebifera, Boxb—FYEYAK.. oe os
Taf. IV, Fig. 5. Lagerstraemia indica, L—Y¥az~y
Taf. IV, Fig. 6. Spiraea betulifolia, Pall—wrAevLEYT 12 os

** —Zweige ziemlich dick.

Taf. IV, Fig. 7. Cladrastis amurensis, Benth et Hook. var. floribunda, Maz.

—{RkDyYy.2 on) teer ws (ie) we Ge me

243

== 243

243
244

. 244

244
244
244

245

SHIRASAWA : UEBERSICHT UND REGISTER. 289

SEITE
Taf. IV, Fig. 9, 10. Cercis chinensis, Bunge —~F+AAY .. «2 oe oe 245

Taf. IV, Fig. 8. Excecaria japonica, ¥. Muell—v FX .. «2 oe oe oe 245
Taf. IV, Fig. 4, Kcelreuteria paniculata, Laxrm.—*# 77 yA «2 «2 oe 245

VII.—Knospen sitzend, von mehreren Schuppen um-
geben. Zweige auffallend schlank, od. besenformig.

A.—Zweige rothbraun glanzend.
a.—Strauchartig.

Lay. Vi, Pig. &. Prunus japonica, Thumb.——=A7 X .. 2. oe s- 22 «+ 245
Taf. III, Fig. 20. Itea japonica, Oliv,—-ZA4+ .. .. «2 «2 «2 «2 © 246
Taf. IV, Fig. 79. Spiraea cantoniensis, Lour.—azv) 2. .. «2 «. «. 246

b.—Baumartig.

Taf. IV, Fig. 72. Betula alba, L. var. vulgaris, Regel —LY 7 Hrs .. .. «. 246
Taf. IV, Fig. 73, rg. Betula Bhojpattra, Wall. var. typica, Sete JF 246

Taf. IV, Fig. 15. Betula alba, L. var. communis, Regel.—¥ ty sn 00 BAG
Taf. IV, Fig. 16. Betula globispica, Shirai—fyYYHY-S .. os 7240
Taf. IV, Fig. zz. Betula alba subsp. latifolia, var. Tauschii.—7 y° 4 ny «. 247
Taf. IV, Fig. 17. Amelanchier asiatica, Koch—Y47IY #7... .. .. «. 247

B.—Zweige grau, dunkelgrau od. graubraun.

Taf. IV, Fig. 18. Pourthiana villosa, Decne—YJJYAIAY.. .. «. 1. os 247
Taf. IV, Fig. 20. Stephanandra flexuosa, Sieb. et Zucc,—aAavAWY 2 247
Taf. IV, Fig. 21. Spiraea japonica, L. fl— DEV 7. «. 0s ee we we 247

Spiraea prunifolia, Sieb. ef Zucc—YVEsF «6 2. 0. 247

VIII.—Knospen sitzend, von mehreren Schuppen umge-
ben, einjahrige Zweige auffallend dick.

A.—Zweige mit Stacheln.

IWS IN, IRS FER INEIEY SG JE — 82 5) Go Oped) un 56 oo ao oo ey ts
Taf. V, Fig. z. Acanthopanax ricinifolinum, Seb. ef Zucc—rI Fy .. .. 248

B.—Zweige ohne Stacheln.
a.—Knospen lang gestreckt, réthlich gefarbt.
Taf. V, Fig. 2. Pirus sambucifolia, Cham. et Sch—t+rAY RF «eve oe 248
b.—Knospen sitzend, griingelb gefarbt.

Taf. V, Fig, 3, . Acanthopanax sciadophylloides, Fr. ef Sav.—a*y €'Y .. 248

IX.—Knospen sitzend, von mehren Schuppen umgeben.
: Knospen stehen an der Spitze der Langtriebe
einzeln, an der Kurztriebe einzeln od. gehauft.

290 SHIRASAWA : UEBERSICHT UND REGISTER.

A.—Zweige mit Stacheln d. h. mit metamorphosierten
Blatt- u. Haargebilden, dornspitzige Zweige nicht
vorhanden.

a.—Je zwei gerade Stacheln neben der Knospe der
Langtriebe.

b.—Gerade einfache od. verzweigte 3-5 spitzige
Stacheln unter Knospen.

SEITE
Ribes grossularia, L.—7YASY—.. .. «2. «- we - 249

Taf. V, Fig. 6. Berberis vulgaris, L—~&€ ) FFA + 249
Taf. V, Fig. 5. Berberis Thunbergii, D.C.— 2% ¥.. = 5249

C.—Mit zwei od. nur einem nach riickwarts gebo-
genen, derben Stachel unter jeder Knospe.
Rosa multiflora, Thuub.— 7 4 737 + 250
Acanthopanax spinosum, Mig—ya*¥ .. . 250

d.—Zahlreiche derbe, zuriickgebogene Stacheln,
ohne Zusammenhang mit den Knospen.

Rubus incisus, Thunb.—%* 4 +2” . 250
B.—Zweige ohne Stacheln (mit od. ohne dornige
Zweige).
a.—Knospenschuppen griin, teilweise braun gefarbt.
a.—Knospen, besonders die unteren Seiten-
knospen klein.
Taf. V, Fig. 9, ro. Ficus erecta, Thunb—4{ XE .. as - 250
Taf. V, Fig. 7, 8. Helwingia ruscifolia, Willd—v\+4HS .. +. 250
8.—Knospen gross od. ziemlich gross.
1.—Knospen, kugelig od. eiformig.
Taf. V, Fig. 11, 72. Spiraea Thunbergii, Siebh—_ HY FAVE .. - 251
2.—Knospen lang gestreckt.
Taf. V, Fig. 27. Populus suaveolens, Fisch—7° .. .. «2 oe oe oo 251
b.—Knospen harzig, glanzend braun.
Taf. V, Fig. 14. Populus tremula, L. var. villosa, Wesm.—¥ ¥+7F7¥Y . 251
Taf. V, Fig. 27. Betula grossa, Sieb. et Zucc—_=ZAX .. Ae on GRE
Taf. V, Fig. 15, 76. Betula ulmifolia, Sieb. ef Zucc—_APYY?2F7NI .. 5 ZG
Taf. V, Fig. 78. Betula Maximwicziana, Regel—Y47\9" 1. -- « Se wie
Taf. V, Fig. 2z. Pirus aria, Ehrh. var. Komaonensis, Wall.—Y 7H ) San van

Taf.
Taf.

Taf.
Taf.

Taf.
Taf.

Taf.

Taf.

Taf.
Taf.
Taf.

Taf.

Taf.
Taf.

Taf.

Taf.

Taf.
Taf.

SHIRASAWA: UEBERSICHT UND REGISTER.

c.—Knospen roth, hell- od. dunkelbraun od. grau-

braun, behaart od. nicht behaart.
a.—Dornspitzige Zweige sind vorhanden.
*.—Knospen behaart.

4 UGS ARS TS Fehon, TUF RG? “BR Go so "cao oo dO
V, Fig. 27. Pirus sinensis, Lindl.—}v..
**, —Knospen nicht behaart.

V, Fig. 23, 24. Mespilus cuneata, Sieb. et Zucc—_YYYY .. .-
Prunus Mume, Sieb. ef Zucc.— a2. om ier
VI, Fig. z. Glochidion obovatum, Sieb. et Zucc—_HyaA)..

8.—Kleine dornspitzige Zweige sind vorhanden.
1.—Blattnarbe halbmondférmig, schmal.
a.—Knospen behaart.

VI, Fig. 2. Lindera glauca, Bl—yYva‘J7Sy ..
VI, Fig. 6. Lindera umbellata, Thunb—fty+7¥) *¥
Prunus persica, Sieb. et Zucc.— € »..
VI, Fig. 9. Prunus tomentosa, Thunb—aZxz7z7YWR.. 56
VI, Fig. 7. Liquidambar Maximowiczii, Mig.—J7'J7.. .. «2

b.—Knospen unbehaart.

VI, Fig. ro. Pirus cathayensis, Hemsl—79 Yy
VI, Fig. rz. Orixa japonica, Thunb—a7v7yv*¥ ..

VI, Fig. rz, Disanthus cercidifolia, Max —~x=avyvyu7.. .. ..
Kerria japonica. Di'C.—V¥ WT .. ce os oo ve
VI, Fig. 3. Lindera triloba, BlL—vyantY .. .. we oe

2.—Blattnarbe rundlich.
*.—Knospen kegel od. spindelformig, etwas gross.

VI, Fig. 137. Windera obtusiloba, Bl—#*y a -84..
VI, Fig. 14, 75. Lindera praecox, BL—SFFIFXY

** —Knospen sehr klein, kugelig.
3.—Blattnarbe dreieckig, schmal.
a.—Knospen behaart.

VI, Fig. 16, 77. Prunus Miqueliana, Max.—t fr vf 79

V, Fig. 25, 26. Pirus Toringo, Sieb. var. incisa, Fr. e¢ Sav.—t 2
+ 255

AeA! oo 60 oo 8p bo oo 00 46 Ga

b.—Knospen unbehaart.

VI, Fig. 79, 20. Prunus Grayana, Max.—Ynrzi X75
VI, Fig. 78. Prunus Buergeriana, Mig.—{x4779..

291

- 253
. 253
» 253
. 253
. 254

+. 254
- 254
» 254

oo As
+ 254

255
5 BR

255

+ 256
- 256

292 SHIRASAWA : UEBERSICHT UND REGISTER.

: : SEITE
Taf. VI, Fig. 21, 22. Prunus cerasoides. Max.—FYVSYIF.. 1. 2. oe 256
Taf. VI, Fig. 23. Prunus Maximowiczii, Rufr—x En F777 omnes Ze

Taf. VI, Fig. 24. Prunns pseudo-cerasus, Lindl. var. spontanea, Maxim.—
ASSIA TAO COM Os es Se Ad oo 55) on BRE
Taf. V, Fig. 79, 20. Pirus Miyabei, Sargent—7 44+Fy.. -. .- «2 «oe 257
Rubus trifidus, Thunb— HY 4 Fa .. 6. we we we we 257

4.—Blattnarbe gross, herzformig.

Taf. VI, Fig. 25. Platycarya strobilacea, Sieb. ef Zuce.—) 7’) ¥ Bo ee
Taf. VI1, Fig. z. Euptelaea polyandra, Sieb. ef Zucce—J¥Y7F Soe 00 St)
X.—Knospen sitzend, von mehreren Schuppen umgeben,

an der Spitze der Langtriebe gehauft.
A.—Knospen nicht von pfriemenformigen Nebenblatt-
chen umgeben.
Taf. VII, Fig. 2. Quercus glandulifera, BlL—a79 .. .. «2 «2 «. c« 257
Taf. VII, Fig. 3. Quercus grosseserrata, BLL—= AFF ~.- +. «. «ws w- 258
Taf. VII, Fig. 4. Quercus dentata, Thunb—By~ .. .. «2 we -- «- 258
Taf. VII, Fig. 5. Quercus serrata, Thunb. var. variabilis, (B/.)—7~¥% .. 258
B.—Endknospen von _ pfriemenformigen Neben-
blattchen umgeben.
Taf. VII, Fig. 6. Quercus serrata, Thunb—7X¥ .. .. oe oe oe eo 258
XI.—Zweige kletternd. (Schlinggewachse).
1.—Knospen sind in der Blattachsel verborgen.
Taf. VII, Fig. 7. Actinidia arguta, Planch—vFFF .. .. «. - 259
Taf. VII, Fig. 8, 9. Actinidia polygama, Mig—Yyre.. .. .. .. «- 259
Taf. VII, Fig. zo. Menispermum davuricum, D.C.—ayeynyF7 .. .. 259
2.—Knospen sind von unausgebildeten, od. zusammen-
gedriickten Blattchen umgeben.
a.—Von unausgebildeten Blattchen.
Taf. I, Fig. 75. Rhus toxicodendron, L. var. radicans, Mig.—'Y# "Pry .. 259
b.—Von zusammengedriickten Blattchen.
Taf. VII, Fig. rr. Cocculus Thunbergii, DC.—74AYNVFTIF -.- «2 «+ 259
3.—Knospen mit einer bezw. zwei Schuppen umgeben.
A.—Zweige ohne Ranken.
Taf. VII, Fig. 15, 16. Berchemia racemosa, Sich. et Zucc—_77¥ +¥ .... 260
Taf. VII, Fig. rz. Wistaria chinensis, Sieb. ef Zucc—_b FYE IF 260
Taf. VII, Fig. 13, rg. Wistaria brachybotrys, Sieb. et Zucc—YVFIF 260

SHIRASAWA : UEBERSICHT UND REGISTER. 293

B.—Zweige mit Ranken.

SEITE
Taf. VII, Fig. 17. Vitis vinifera, L—7o°'7 we. Sale itegoe Gan se ae 200
Taf. VII, Fig. 18. Vitis flexuosa, Thunb—¥ aYrox¥wJzy.. 50 om AS
Vitis Thunbergii, Sieb. ef Zucce—meEY wr so to Aan
4.—Knospen von mehreren Schuppen umgeben.
a.—Knospen spindelférmig.
Taf. VII, Fig. 19. Schizandra chinensis, Baill—7Yevatzy .. .. .. 261

8.—Knospen kugel- od. eiformig.

Taf. VII, Fig. 20. Akebia quinata, Decne.—77E So bey on cn Go Hehe
LajVil,, biz. 21. Akebia lobatay Decne—= 7 70, FE wa os ei) ae we ZOE
Taf. VII, Fig. 22. Celastrus articulatus, Thunb.— ‘yr x*® RY .. «2 oe 261

294 SHIRASAWA: UEBERSICHT UND REGISTER.

TABELLE II.

Knospen stehen an der Langtriebe abwechselnd,
“* Zweizeilig.”

I.—Knospen (Bliit- u. Laub-) von wenigen Schuppen
umgeben.
A.—Knospen kugelig, od. eiformig.

SEITE
Taf. VIII, Fig. r. Colylopsis pauciflora, Sieb. et Zucc.—e YHA .. «. 262

B.—Knospen spindel- od. kugelfGrmig.
Taf. VIII, Fig. 2. Corylopsis spicata, Sieb. ef Zucc.— FYE KX .- «2 we 262
C.—Knospen etwas flach.

Taf. VIII, Fig. 3, 4. Stuartia monadelpha, Sieb. et Zuce.—yny 50° bo AEB
Taf. VIII, Fig. 5. Stuartia pseudo-Camellia, Max.—}‘Y n-S% .. «ee 262

II.—Knospen kugelig od. keil- od. eiformig mit zwei od.
nur wenigen Schuppen.
A.—Knospen dusserlich von einer grossen umfassen-
den u. einer kleineren Schuppe umgeben.
Taf. VIII, Fig. 6, 7. Tilia Miqueliana, Max,—74{y2 .. . 5 ae Han)
Taf. VIII, Fig. 8. Tilia cordata, Mill. var. japonica, Migea 5 aa 203)
B.—Knospen von mehreren Schuppen umgeben.
a.—Knospen behaart.

Taf. VI, Fig. 9. ‘Celtis sinensis, Pevo—s2 J %.. <5 <2 <5 = «ml se 203

Taf. VIII, Fig. r2, 73. Corylus heterophylla, Fisch—-\ysS } .. «2 .«. 263

Taf. VIII, Fig. rz. Corylus rostrata, Ait. var. Sieboldiana, Max.—'Y ) -vv
TS “Ree eG a OG OO Os oo FERIA.

£.—Knospen unbehaart.
Taf. VIII, Fig. rg. Castanea vulgaris, Lam. var. japonica, DC.—7 Y o. 264

II1I.—Knospen spitz, walzenformig, spindel- od. kegelfor-
mig, mit zahlreichen Schuppen.
A.—Schuppen spiralig angeordnet.
a.—Knospen spindelformig.
Taf. VIII, Fig. 16, 77. Fagus sylvatica, L. var. Sieboidii, Max.— 7+ .. 264
Taf, VIII, Fig. 15. Fagus japonica, Max.— 4X7 F.. .. «2 os oe co 204

£.—Knospen kegelformig.

Taf. IX, Fig. 1, 2. Wex macropoda, Mig.—J7F4".. .. «. «2 oe c+ 265
Taf. IX, Fig. 3, ¢- Ginkgo biloba, L.—FFY .. .. ss oe =- se veo 205

SHIRASAWA: UEBERSICHT UND REGISTER. 295

y.—Knospen spitz, walzenformig.
Taf. IX, Fig. 15, 16. Ostrya virginica, Willd—7 US" .. «2 «2 8 oe 205
B.—Schuppen abwechselnd angeordnet.

g.—Schuppen auf zwei Seiten angeordnet.
*.—Knospen behaart.

Taf. VIII, Fig. ro. Aphananthe aspera, Planch—»7)% .. «2 «. « 265
** —Knospen gar nicht, oder wenig behaart.

Taf. IX, Fig. 5, 6. Ulmus campestris, Sm. var. vulgaris, Planch.—=a 7
a 3) Bb Sol NOUS eIOae toe ence aon mon Homey
Ulmus parvifolia, facg—F+#=ov .. .. «2 «. «- 266
Taf. IX, Fig. 8, 9. Ulmus campestris, Sm. var. levis, Planch._—/\r =v .. 266
Taf. IX, Fig. 7. Ulmus montana, Sm. var. laciniata, Trautvu—J'Yy.. .. 266

8.—Schuppen von vier Seiten abwechselnd an-

geordnet.
Taf. IX, Fig. zo. Carpinus japonica, Bl—7Yvsz .. .. .. 2. 0 «- 266
Raf, IX, Fug. a, Carpinus| yedoensis, Maz—4xX 27° .. .. «. «e + 200
Taf. IX, Fig. 13. Carpinus laxiflora, Bl—yna ae) ate) ai ates wie ZOO
Taf. IX, Fig. rz. Carpinus cordata, BL—YaAywN” .. .. 1. «2 «2 o- 267
Taf. IX, Fig. 14. Zelkowa acuminata, Planch—y¥*% .. .. «os «+ « 267

IV.—Knospen von zusammengedriickten Blattchen ge-
bildet (nackt).

Taf. IX, Fig. 19. Picrasma quassioides, Benn.—=jff% .. ..' «2 «2 «+ 267
Taf. IX, Fig. 17, 18. Hamamelis japonica, Sieb. et Zucc.—Y v7 .. .. 267

296 SHIRASAWA: UEBERSICHT UND REGISTER.

TABELLE III.
Knospen ‘“ gegenstandig.”’

I.-—Knospen sind in der Blattnarbe verborgen.

SEITE
Taf. X, Fig. 1, 2. Philadelphus coronarius, L. var. Satumi, Mazx.—,*4 7
2D Tie RCC MRO OG SOOT adie o¢ 46. 50 ‘Zit
II.—Zweige auffallend dick, Endknospe sehr gross.
Taf. X, Fig. 3. Aesculus) tucbinata, Bl.—p} 3-9) 2.) 22) oe eee eas
III.—Zweige sind zumeist dornspitzig.
Taf. X, Fig. 6, 7. Punica Granatum, L.—+#F¥ 7 n soll Eaeied Cale) (elena CEE ZOS
Taf. X, Fig. 4.5. Rhamnus japonica, Max. var. genuina, Max.—7unY x
oil a oe SC Te OOF amEerON «td o-oo a0 Zee

IV.—Knospen von unausgebildeten Blattchen, nicht von
eigentlichen Schuppen umgeben.
A.—Knospen gestielt.

a.—Zweige (zweizeilig angeorduet) grau gefarbt.

Taf. X, Fig. 8. Callicarpa mollis, Sieb. ef Zucc—¥ FLFYUH .. «. «. 269
Taf. X, Fig. ro. Callicarpa japonica, Thunb—aFoRLDET .. -- «. 269
Taf. X, Fig. 9. Callicarpa purpurea, $uss—ALFYUX .. «2 «2 2. «. 269

b.—Zweige braun gefarbt
Taf. X, Pig. 1, 12. Niburnum~furcatum, BL — 27939) else zOO)

B.—Knospen sitzend an der Triebe.

Taf. X, Fig. 137. Clerodendron trichotomum, Tiunb.—7Y¥¥.. .. .. «. 269
Taf. X, Fig. 14, 15. Cornus ignorata, Koch.—tari XX... .. «02 «. 0 269
Taf. X, Fig. 16. Premna microphylla, Turcz.—arv 74 .. .. «. .~. 270
Taf. X, Fig. 77. Vitex Negundo, E.—=Y sy ss. oes ee) eee zzO

Vitex trifolia, L. var. unifoliolata, Schauer.—~.v a°"J .. 270
Taf. X, Fig. 18, 19. Evodia rutecarpa, Benth et Hook—a*ya .. .. «. 270

V.—End- od. Seitenknospen so von zwei Schuppen dicht
umschlossen, dass scheinbar nur eine Knospen-
schuppe vorhanden ist.

A.—Knospen unbehaart, glanzend.

a.—Knospen gestielt.

Taf. X, Fig. 20. Viburnum Opulus, L.—wyaAZ.. .. ». «2 «2 «os os 270

Taf.
Taf.

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Taf.
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.

SHIRASAWA : UEBERSICHT UND REGISTER.

X, Fig. 22. Acer rufinerve, Sieb. et Zuce— JY HAF 2.2 . os

X, Fig. 23, 24. Acer crategifolium, Sieb. et Zucc— AVY HaF.. «. 271
b.—Knospen sitzend.
X, Fig. 2z. Staphylea bumalda, Sieb. et Zucc.— = ‘Y/S'V'Y% os «2 oe 271
X, Fig. 25, 26. Acer palmatum, Thunb—jn~F5.. .. .. «- 00 PYA
III, Fig. 7. Salix multinervis, Fr. et Sav.—ay+~ + ..* .. 30 Arh
Uh, Wes 7, Rebs suily ey Joss LV SING SPasan son 60 oo on oo oO yf
B.—Knospen behaart.
X, Fig. 28. Phellodendron amurense, Rupr.i—¥ -H*.. 6. we we 272
IG eS AT INES! CRSVONOIN, IGG 2 Wispeitess~ Go do an ao do ye
X, Fig. 30, 97. Cornus Kousa, Buerg.—V¥V#YY .- «2 2 of «. 272
X, Fig. 29. u. Taf. XI, Fig. z. Cornus officinalis, Sieb. ef Zucce.—¥ vy
MSL Sod ao 6d 05 050 oc 272
VI.—Seitenknospen eiformig od. spindelig, fast senkrecht
abstehend.
XI, Fig. 4. Lonicera Morrowii, A. Gray.—%+y¥VAi7.. .. «2 «- 272
XI, Fig. 2, 3. Lonicera gracilipes, Mig.—9 742% ee So ca oo 27/5)
Lonicera cerulea, L—V7H2E)YF{AZNFF .«.. .. 273
VII.—An den Seitenknospen nur wenige Schuppen, 2 od.
3 sichtbar.
A.—End- u. Seitenknospen kantig.
a.—Knospen od. Zweige behaart, od. klein walzig.
XI, Fig. 5. Viburnum dilatatum, Thunb.—fryv Az .. «2 2. oe «+ 273
Abie. 7. Naburnum~ Sieboldi, Migr ¥ 2. ie ee ee ee 5 FE
XI, Fig. 6. Viburnum tomentosum, Thunb.—¥ VF VY .. «- oe oe 273
XI, Fig. 8, 9. Viburnum Wrightii, Mig—i¥V¥i¥X3 .. «. +. 274
b.—Knospen nicht od. ein wenig behaart.
XI, Fig, 72. Fraxinus Sieboldiana, Bl—y a Fo. «2 oe ee we oe 274
XI, Fig. ro, rz, Fraxinus Bungeana, D.C. var. pubinarvis, Bl.—
Spe) oo bo * Sash Rereletelss Uielol ete) 274
XI, Fig. 13. Fraxinus longicuspis, Sieb. et Tc ae FAY) a4 - 274
XI, Fig. zg. Chionanthus retusa, Lindl. et Paxt.—t bY 7a" .. «2 274
B.—Seitenknospen angedriickt, an der dem Zweige
zugewendeten Seite abgeplattet.
a.—Knospen u. Zweige nicht behaart.
a.—Blattnarbe schmal.
XI, Fig. 17, 78. Acer argutum, Maz.—F¥)JrnAF .. TemrSIsn2 75
XI, Fig. 15, 16. Acer distylum, Sieb. et Zucc.—t bY -SHAF 2. os 275

XI, Fig. 79. Cercidiphyllum japonicum, Sieb. ef Zucco—H'VYF .. «

298 SHIRASAWA: UEBERSICHT UND REGISTER.

b.—Blattnarbe gross, rundlich od. halbmondformig.
* —Baumgewéachse.
Taf. XI,- Fig. 20. Euscaphis japonica, Pax.—a*'yK4 .. .. «2 oe o- 275
Taf. XI, Fig. 2z. Acer japonicum, Thunb.—AYFAHAF 2. «2 oe «- 276
Acer Sieboldiana, Mig —anJFANHAF 2. «2 22 oe 276
** —Strauchgewdachse.

Taf. XI, Fig. 23. Hydrangea hortensia, Smith. var. ee Maxim.—

Ww7. 50 © | ee) we Ss e270
Hydeances Thunbergii, ‘Sicb. _7 wy FRc See 8 ae OTE
Taf. XI, Fig. 24. Hydrangea hortensia, DC.—7 F444 .. .. «. «. «. 276

b.—Knospen u. Zweige behaart.
Taf. XI, Fig. 25. Ligustrum Ibota, Sieb.— 4 #4 ) ¥ wis) ele! woiuictel tae TO

VIII.—An den Seitenknospen mehr als drei Schuppen
sichtbar.
A.—Knospen u. Blattnarbe gross.
a.—Knospen kugelig.

Sambucus racemosa, L. var. Sieboldiana, Miqg.—=7) f 3. 277

b.—Knospen kantig.
Taf. XI, Fig. 28. Syringa vulgaris, L—aZyeryvEf.. 2. «2 «2. «- 277
Taf. XI, Fig. 27. Syringa japonica, Maxy.—aAy 4 .. .. «. 06 os we 277,
B.—Knospen weniger gross, von den Schuppen sehr
lose umhiillt od. nicht. Blattnarbe ziemlich gross.
a.—Knospen kegel- od. spindelig.
a,—Knospen unbehaart.
Taf. XII, Fig. 1. Evonymus oxyphyllus, Mig.—'y yaxy.. .. 2. e+ 0 297

Taf. XII, Fig. 3. Forsithya suspensa, Vahl—vy¥49.. .. .. mere
Taf. XII, Fig. 2. Viburnum phlebotrichum, Sieb. et Zucc.—7 b a4 oe

8.—Knospen behaart.
Taf. XII, Fig. 4. Acer nikoense, Max.—F-3Y2¥ )¥ «- 2s 22 oe oe 278
b.—Knospen kantig.
a.—Knospen von eigentlichen Schuppen umgeben.

Taf. XII, Fig. 5, 6. Acer carpinifolium, Sieb. ef Zucc.—FFY 7% «2 «+. 278
Taf. XII, Fig. 7. Acer Ginnala, Max.—HFAR.. oe oe oe 2s oe oe 279
Taf. XII, Fig. 8. Evonymus alatus, Fr. et Sav—=ay%W .. «oe 2+ 278
Taf. XII, Fig. 9. Evonymus europeus, L. var. Hamilitonianus, Maz.—
SfSsS 56 > Bae ICO oo. On Onkol SE
Taf. XII, Fig. zo. Deutzia scabra, Thunb. ane ye aie. ei. whe, | nee Tete eee 7G
Taf. XII, Fig. 15. Calycanthus precox, L.—F 754... oe ee 2+ oe os 278
Taf. XII, Fig. 73, 4. Diervilla grandiflora, Sieb. et Zucc. —naF FY ¥ oe 279

-

SHIRASAWA : UEBERSICHT UND REGISTER. 299

Taf. XII, Fig. rz. Diervilla japonica, Sieb, ef Zuec—pAayy¥ .. ».. «- 279
Taf. XII, Fig. 26. Ligustrum medium, Fr. et Sav.—4F w&rS 4 HY IX. -. 279
f8.—Knospen von pfriemenformigen Schuppen
umgeben.
Taf. XII, Fig. rz. Deutzia gracilis, Sieb. ef Zucc—te XYYE «2. «2 «+ 279
c.—Knospen kugelig.
Taf. XII, Fig. 77. Catalpa Kempferi, Sieb. ef Zucc.—%¥ Ur .. «2 «. 280
Taf. XI, Fig. 22. Hydrangea paniculata, Siebh— 7) yD ¥.. «2 «2 2 « 280
C.—Knospen an der Spitze der Triebe gross, an der
Basis klein.
a.—Blattnarbe klein.
Taf. XII, Fig. 19. Acer pycnanthum, C. Koch—”}+HA5 .. .. .. «. 280
b.—Blattnarbe ziemlich gross.
Taf. XII, Fig. 20. Acer purpurascens, Fr. et Savu.—jFfU~FK .. .. «. 280

Taf. XII, Fig. 18. Acer pictum, Thunb—{ pvr H~z .. 2 «2 «2 «. 280
LN FSAI, IU, Wels B OB do nd co og oa op eli

D.—Knospen klein.
a.—Knospenschuppen gegenstandig angeordnet.
a.—Zweige diinn.
Taf, XII, Fig. 21. Abelia serrata, Sieb, ef Zucc.—saYIWNBAYGY¥ .. .. 281
#.—Zweige auffallend dick.
Taf. XII, Fig. 16. Paulownia tomentosa, Baill.—* Y Sc sin on gg PASI

b.—Knospenschuppen spiralig angeordnet.

IX.—Zweige kletternd (Schlinggewachse).
1.—Knospen sind von einer od. zwei Schuppen
umgeben.
Gio 290, Tie, A PESTA TCC, Jo PPG agiog GO 08 oo On on Zieh

2.—Knospen von mehreren Schuppen umgeben.
A.—Zweige mit Ranken.
Taf. XII, Fig. 23. Clematis japonica, Thunb—YyYRRUYF .. .. «. 281

B.—Zweige ohne Ranken.

Taf. XII, Fig. 24. Schizophragma hydrangeoides.— 4 rif 7 = .. «. «. 282
Taf. XII, Fig. 25. Wydrangea petiolaris, Sieb. et Zucc. var. cordifolia, pe
STROLL gg c's acest pe~ Malan el eer Sa

300

Taf.

Taf.
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Taf.

Taf.
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Taf.

SHIRASAWA: UEBERSICHT UND REGISTER.

TABELLE IV.
Zweige “ quirlstandig.”

I.—Knospen sind von unausgebildeten Blattchen um-
geben.

SEITE
XIII, Fig. 1, 2. Clethra barbinarvis, Sieb. ef Zucc—Y3AYP .. .. 283

I].—Endknospe sehr gross, einjahrige Zweige auffallend
dick.

XIII, Fig. 4. Sterculia platanifolia, L—f4t¥y .. .. .. « .. 283
XII, Fig. 3. Idesia polycarpa, Maz.—4 FF ¥Y.. we oe we ee weee8g
AU, Fig. 5. Aleuritesiicordata, D).C.— 7, Jaz 4a) ease ssl oS

III.—Knospen sind von mehreren Schuppen umgeben,
an der Langtriebe einzeln od. gehauft.

A.—An der Langtriebe einzeln.
a.—Knospen kugel od. eiformig.

XIII, Fig. 6, 7. Cornus macrophylla. Wall.— = K*%.. .. .. «. «2 283
XIII, Fig. 8, 9. Andromeda cernua, Mig.—my77'yYrny .. .. .. 284

b.—Knospen spindelig.
a.—Knospenschuppen behaart.
XIII, Fig. ro, rz. Rhododendron dilatatum, Mig.— = 'Y7*Yny.. .. 284
8.—Knospenschuppen unbehaart.

XIII, Fig. 12. Rhododendron sinense, Sw.-¥ UY A YrNY .. .. «. 284

XIII, Fig. 73, 74. Enkyanthus japonicus, Hook.—FYSZ*y .. .. «. 284
B.—An der Langtriebe gehauft.

XIII, Fig. 15. Rhododendron Schlippenbachii, Max.—7H7IA'YrY.. 284

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Untersuchungen ueber das Klemmen der
technisch wichtigsten Japanischen Holzarten,
VON

F. Koide, Ringakushi.

I. Einleitung.

Dass die genaue Kenntniss der technischen Eigenschaften
der Holzer so gut fiir den Forstmann, als fiir den holzver-
brauchenden Techniker von der grossten Wichtigkeit ist,
unterliegt keinem Zweifel. Indessen kommen Forstleute und
Holzconsumenten selten in unmittelbare Beriihrung, was zur
Folge hat, dass die fachlichgebildeten Forstleute oft nicht zu
beurtheilen wissen, welche Ejigenschaften das von ihnen ge-
lieferte Holz besitzt. Es ist dies mit der Hauptgrund, warum
die Forstbenutzungslehre noch so wenig Fortschritt gemacht
hat.

Besonders in Japan, wo das Holz eine viel ausgedehntere
und vielseitigere Verwendung findet, als in anderen Landern,
darf diese Kenntniss fiir den Forstmann in dem Grade, wie fiir
den holzverbrauchenden Techniker als unentbehrlich bezeichnet
werden. Hs liegt daher uns japanischen Forstleuten, ob uns
mehr mit der Ausbildung der Forstbenutzungslehre zu beschaf-
tigen, als es von den europaeischen Fachgenossen bisher gesche-
hen ist.

Wenn man im Allgemeinen die Eigenschaften der Hélzer
betrachtet, welche auf der Zusammensetzung und mittelbar
auf den dusseren Lebensumstanden des Baumes beruhen, so
kann man folgende drei Gruppen unterscheiden.

1. Higenschaften nach der dusseren Erscheinung, welche
durch den Gesichts- Geruchs- und Tastsinn wahr-
nehmbar sind.

2. Eigenschaften, welche den materiellen Zustand dar-
stellen.

302 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

3. Ejigenschaften nach dem Verhalten gegen von aussen

einwirkende Krafte.

Da von diesen Eigenschaften besonders diejenigen, welche
in die zweite Gruppe fallen, und von diesen wieder das, wie
ich glaube, zuerst von Nordlinger genauer untersuchte Klemmen,
und Schwinden von hoher Wichtigkeit fiir die Bearbeitung des
Holzes fiir Gewerbe sind, so sollen im Folgenden diese beiden
Eigenschaften, wenn auch einiger der technisch wichtigsten
japanischen Holzer, naher untersucht werden.

II. Untersuchungsmaterialien.

Wenn man von einem frischgefallten Stamme eine Scheibe
abschneidet, und an derselben in radialer Richtung vom Umfang
gegen dem Mittelpunkt sagt, so nahern sich die Sagschnitts-
flachen in Folge der Spannungskraft des Holzkorpers.

Diese Erscheinung nennt man bekanntlich das Klemmen
des Holzes und die Untersuchung dieser Erscheinung ist
von besonderem Interesse, weil sie einen sehr grossen Einfluss
alissert

1. Auf die Schwindungsgrosse,

2. Auf die Saégearbeit der Holzhauer und endlich,

3. Aut den Ueberwallungsprocess der Spiegelrisse im

lebenden Baume.

Den Grundstock meines Untersuchungsmateriales bilden
die in der Gegend von Chichibu und Nikko gefallten Baume.

Das Gebiet, in welchem die Nadelholzer Larix leptolepis,
Pinus densiflora, Abies Umbellata, Cryptomeria japonica,
und Chamaecyparis obtusa, die winterkahlen Laubholzer,
Betula alba var, vulgaris, Magnolia hypoleuca, Prunus
cerasoides, Zelkowa keaki und Juglans Sieboldiana gefallt
wurden, ist derjenige Theil des Mitsuminegebirges, welcher
Aza Isedaira genannt wird und einen Theil des Chichibu-
reviers bildet. Die Fallungszeit war Anfang April. Die fiinf
Stamme von Tsuga Sieboldii, Betula ulmifolia, Acer Pal-
matum, Fagus japonica und Halesia corymbosum wuchsen

KOIDE: UEBER DAS KLEMMEN DER HOELZER. 303

in der Nahe von Nikko und zwar in der Privatwaldungen
Aza Umakayeshi und wurden am Anfang Mai gefallt.

Die vorgenannten Untersuchungsmaterialien dienten nun
in erster Linie zur Feststellung der Grosse des Klemmens
bei den verschiedenen Holzarten und ferner dazu, zu ermitteln,
welchen Einfluss auf die Grosse des Klemmens die Rinde
des Baumes, die Dicke der Scheibe, die Lange des Sagschnitts
und der eigentliche Splinttheil &c., ausiiben und endlich zu
dem Versuche, den Grad des Klemmens als eine Function des
Abstandes vom Mittelpunkt der Scheibe darzustellen.

Das Untersuchungsmaterial bestand aus:

Larix leptolepis (Karamatsu).

Auf einem Berghange mit einer Neigung von ca. 25°

und der Exposition nach SW wurde ein 50-jahriger Baum
gefallt, welcher hier bei geniigendem Schlusse erwachsen war.
Seine ganze Hohe betrug 23,4 m., die Lange des astfreien
Schaftes 15 m. und der durchschnittliche Durchmesser 28,5 cm.
in Brusthohe. Der Stamm war vollholzig und der Jahrring-
bau schon concentrisch.

Pinus densiflora (Akamatsu).

Lage, Neigung, Exposition und Schlussverhaltnisse waren
ganz ahnlich wie bei Larix leptolepis.

In einem gemischten Bestande von Sugi und Hinoki mit
einzelnen Akamatsu liess ich eine der letzteren fallen. Die
ganze Hohe des 82jahrigen Stammes betrug 20,5 m. die
Lange des astfreien Schaftes 11,5 m. und der durchschnittliche
Durchmesser in Brusthohe 33,5 cm. Der Stamm war voll-
holzig und der Jahrringbau ziemlich concentrisch.

Abies umbellata (Uvashivomomzt).

In derselben Lage wie Akamatsu wurde ein 82jahriges
Exemplar gefallt.

Die ganze Hohe des Baumes betrug 18,4 m, die Lange
des astfreien Schaftes 7,2 m. und der durchschnittliche Durch-

304 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

messer 31,5 cm. in Brusthéhe. Der Stamm war ‘sehr voll-
holzig und der Jahrringbau schén concentrisch.

Cryptomeria japonica (Swg7).

Auf einem Hange mit einer Neigung von ca. 30° und
der Exposition nach SO wurde ein Probestamm von 75
jahrigem Alter, einer GesammthGdhe von 21 m einer Lange des
astfreien Schaftes von 12,5m., einen durchschnittlichen Durch-
messer von 25,3cm in Brusthdhe ausgewahlt. Der Stamm
war in einem mit Hinoki und Sawara gleichmassig gemischter
Bestande im dichten Schlusse erwachsen. Der Stamm war
auch vollhdlzig und der Jahrringbau schon concentrisch.

Chamaecyparis obtusa (Hinoki).

Ein unter denselben Verhaltnissen mit der Sugi erwach-
sener Baum wurde gefallt. Das Alter war 88 Jahre, der
Stamm vollholzig und der Jahrringbau concentrisch.

Die ganze Lange des Stammes betrug 24 m., die Lange
bis zum Kronenansatz 13,5 m. und der durchschnittliche
Durchmesser in Brusthohe 23 cm.

Betula alba var. vulgaris (Siirakaba).

Ein Baum von 33 jahrigem Alter wurde auf einem Hange
einer Neigung von ca. 30° mit einer nordlichen Exposition
gefallt, wo er im gentigenden Schlusse erwachsen war. Die
ganze Hohe des Baumes betrug 18 m, die Lange des astfreien
Schaftes 7 m. und der durchschnittliche Durchmesser in
BrusthGhe 23cm. Der Stamm war sehr astreich und der
Jahrringbau etwas excentrisch.

Magnolia hypoleuca (Honok:),

Aus einem mit verschiedenen anderen winterkahlen Laub-
hélzern gemischten und sehr dicht geschlossenen Bestande,
auf dem Hange einer Neigung von ca. 30° und der Exposition
OS. Das Alter war 65 Jahre, die ganze Hohe des Baumes
betrug 17,5°m, die Lange des astfreien Schaftes 8,5 m. und
der durchschnittliche Durchmesser in Brusthohe 32 cm, Der

KOIDE: UEBER DAS KLEMMEN DER HOELZER. 305

Stamm war sehr astreich und abholzig und der Jahrringbau
dabei sehr excentrisch.

Prunus cerasoides (Mejivo-zakura).

Ein Probestamm von 52jaéhrigem Alter wurde in einem
wie bei Honoki gemischten, aber etwas lichten Bestand
ausgesucht.

Die Neigung des Standortes betrug ca. 10, und die Ex-
position war Siid. Der Stamm besass eine Gesammthohe
von 16,5 m, eine Lange des astfreien Schaftes von 6,5 m.
und einen durchschnittlichen durchmesser in Brusthohe von
22,5cm. Der Stamm war sehr astreich und der Jahrringbau
daher sehr excentrisch.

Zelkowa Keaki (Keak).

Auf einem Berghange mit einer Neigung von ca. 30°
und einer westlichen Exposition fand ich eine 82 jahrige
Keaki, welche in gemischtem Bestande von Sugi und Hinoki
im ziemlich dichten Schlusse erwachsen war.

Die ganze Hohe des Stammes war 20,6m, die Schafts-
lange bis zum Kronenansatz 8,1m, und der durchschnittliche
Durchmesser in Brusthohe 35 cm. Der Stamm war sehr
astreich, aber der Jahrringbau ziemlich concentrisch.

Juglans Sieboldiana (Om -guruit).

Auf einem Hange mit einer Neigung von ca. 50° und
der Exposition SW wurde, unter ahnlichen Bestand-verhalt-
nissen wie bei Mejiro-zakura angegeben wurde, eine 70 jahrige
Oni-gurumi mit einer gangen Hohe von 13,5 m, einer Linge
des astfreien Schaftes von 5,7 m und einem durchschnittlichen
Durchmesser in Brusthohe von 28,5 cm gefallt. Der Stamm
war etwas kranklich, sehr astreich und abholzig und der
Jahrringbau sehr excentrisch.

Tsuga Sieboldii (Tsuga),

In einem Bestande mit einer Neigung von Ca. 20° und
einer nordwestlichen Exposition wurde ein Baum von 80
jahringem Alter, einer Gesammthéhe von 16 m, einer Lange

306 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

des astlosen Schaftes von 4 m und einem durchschnittlichen
Durchmesser in Brusthédhe von 25,4 cm. gefunden.

Der Baum war in gemischtem Bestand von verschiedenen
anderen winterkahlen Laubhélzern in Einzelmischung. Aus
dem Verhaltnisse des Starkezuwachses schien es, dass er von
zwanzigstem Jahre bis elwa vierzigstem durch andere Baume
stark untergedruckt wurde und er spater in Folge giinstigerer
Schlussverhaltnisse wieder kraftiger gewachsen ist. Der Stamm
war sehr astreich und abholzig, aber der Jahrringbau ziemlich
concentrisch.

Betula ulmifolia (Yoguso-minebart),

In einem mit verschiedenen Laubholzern gemischten und
beinahe gentigend geschlossenen Bestande auf einem Hange
von einer Neigung von ca. 20° und bei siiddstlicher Exposition
wurde ein 65jahringer Baum gefallt. Die ganze Hohe des
Baumes betrug 12m, die Lange des astlosen Schaftes 7m
und der durchschnittliche Durchmesser in Brusthéhe 26 cm.
Der Stamm war sehr astrein und ziemlich vollhdlzig und
der Jahrringbau ungefahr concentrisch.

Acer palmatum (Momzjt).

In demselben Standort mit der Yoguso-minebari fand ich
auch eine 6ojahringe Momiji, welche daher unter denselben
Verhaltnissen erwachsen war. Die ganze Hohe des Baumes
betrug 12m, die Lange des astfreien Schaftes 4m. und der
durchschnittliche Durchmesser in Brusthodhe 30,2 cm.

Der Stamm war sehr astreich und der Jahrringbau
excentrisch.

Fagus japonica (Jnu-buna),

Unmittelbar an dem vorigen Bestand angrenzend und
zwar bei einer Neigung von ca. 30° und einer stidwestlichen
Exposition fand sich auch eine 60j4hrige Inu-buna, die unter
ihnlichen Schlussverhaltnissen erwachsen war, wie die beiden
vorgenannten Stamme. :

Die ganze Hohe des Stammes betrug 12m, die Lange
des astlosen Schaftes 3,5 m und der durchschnittliche Durch-

KOIDE: UEBER DAS KLEMMEN DER HOELZER. 307

messer in Brusthohe 24cm. Der Stamm war sehr astreich
und der Jahrringbau etwas excentrisch.

Halesia Corymbosum (Asagara),

Auf einem Hange mit einer Neigung von ca. 25° und
einer nordostlichen Exposition wurde cine 5o0jahrige Asagara
gefallt, welch von Jugend auf etwas licht erwachsen war.
Der Stamm war sehr astreich und der Jahrringbau sehr
excentrisch. Die Gesammthohe betrug 13 m, die Schaftslange
bis zum Kronenansatz 5m und der durchschnittliche Durch-
messer in Brusthéhe 21,5 cm.

III. Methode der Untersuchung

des Klemmens.

Es ist selbstredend, dass, wenn die Resultate einer Unter-
suchung mit Ergebnissen der von Anderen angestellten
ahnlichen Untersuchungen iiber einen Gegenstand verglichen
werden sollen, man wohl denselben Weg einschlagen muss,
wo es sich nicht darum handelt die Constanz einer Erscheinung
festzustellen. Bei der Untersuchung tiber das Klemmen habe
ich aber nicht immer denselben Weg inne gehalten, wie z.B.
Nordlinger vorgeschrieben hat. Da ich wahrend der Versuche
gemachte Erfahrungen zur Verbesserung der Verfahrungs-
weise bei spdteren Experiementen verwendet habe, liegt mir
nun ob das von mir beobachtete Verfahren zu schildern.

Unmittelbar nach der Fallung der Baume entnahm ich
von den einzelnen Stémmen 4 oder 5 Scheiben ca. I cm.
Dicke in verschiedenen Baumhohen und in den nach Siiden
gerichten Theil der Scheibe liess ich (Siehe Fig. I und II)
eine Kluft in einer Breite von 1 cm. vom Umfang bis zum
Mittelpunkt ausschneiden und dann mass ich die Abstaénde, um
welche je zwei zu beiden Seiten der Kluft befindliche Punkte
zusammengeriickt sind.

308 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

Fig. I. Fig. Il.
k Kernholz. zt Innerer Theil.
z Zwischenzone. m Mittler Theil.
s Splintholz. a Aeusserer Theil.

Der schraffierte Theil der Figuren zeigt den Sagschnitt
und 1-1, 2-2 und 3-3 sind die gemessenen Entfernungen.

Bei Holzern wie Akamatsu und Karamatsu, welche
einen~ deutlichen Farbenunterschied zwischen dem Kern
und Splint zeigen, habe ich das Klemmen in Splintholz,
Zwischenzone und Kernholz und zwar im Mittelpunkte
jeder zone, wie in Fig. I. gemessen.

Bei Urashiro-momi, Shirakaba u. dgl., wobei der dussere
und innere Theil dieselbe Farbung zeigen, habe ich das
Klemmen im ausseren, mittleren und inneren Theile mit
einer bestimmten Entfernung vom Mittelpunkt gegen den
Umfang gemessen.

Der Einfluss der Rinde auf das Klemmen wurde durch
Untersuchung der in fast gleichen Hohen tiber dem Boden
sich befindlichen d.h. dicht neben cinander abgesagten Schei-
ben festgestellt, in denen die eine mit Rinde und die andere
ohne Rinde blieb.

Das Verhaltniss des Klemmens zur Lange des Sdagsch-
nitts wurde besonders durch Untersuchung der Scheiben von
Urashiro-momi und Karamatsu klargelegt.

Bei dem Material, welches ich in der Nahe von Uma-
kayeshi nahm, wurde einige Abweichungen von dem _ vorer-
wahnten Verfahren vorgenommen in Bezug auf die Ziehung
der Grundlinien und zwar habe ich statt der dreieckigen
Form zwei Parallel linien mit der Distanz von 3 cm von
einander vom Mittelpunkt gegen die Rinde wie Fig. III.
und IV.

KOIDE: UEBER DAS KLEMMEN DER HOELZER. 309

eo
Ss
Fig. IV.

Um die Beziehung des Klemmens zur Dicke der Scheiben
zu ermitteln, wurde zunachst 1m von dem unteren Schnitt-
rande des Stammes, also in ungefahr 1,3m Hohe tiber
dem Boden 5 Scheiben mit einer allmahlich zunehmenden
Dicke 1, 2, 3, 4 und 5 cm von jeder Holzart herausgesch-
nitten und in der ndmlichen Weise untersucht.

Um zu priifen, ob das Klemmen in der Linie eines
einzigen Sadgschnitts des Gesammt-klemmen im ganzen Um-
fang der Scheibe ausdriicke, wurden Scheiben von Tsuga,
Yoguso-minebari, Momiji, Inu-buna und Asagara zur Unter-
suchung benutzt.

Diese Scheiben wurden in 2m Hodhe vom Fuss entnom-
men und an jeder derselben wurden vier Sagschnitte nach
der Himmelrichtung von 7 cm Lange gegen dem Mittel-
punkt zugefiihrt wie es in Fig. V. zeigt.

Ss
Fig. V. Fig. VI. Fig. VII.
Eigentlicher Splintring. Eigentlicher Kerntheil.

Der schrafherte Theil der Figuren zeigt dem Sa&gschnitt
und I-I, 2-2, sind die im Splint oder dusseren Theil bei
Fig. V. und VI. und in der Mitte des Kerntheiles bei Fig.
VII. gemessenen Entfernungen.

310 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

Die Grosse des Klemmens von eigentlichem Splint und
Kernholz fiir sich, untersuchte ich an Scheiben von Asagara
und Yoguso-minebari, aus welchen je ein Splintring und
Kerntheil (aus einer und derselben Scheibe wie in Fig. VI.
und VII. dargestellt) herausgearbeitet worden waren.

IV. Resultate der Untersuchung.

Die nach den so beschriebenen Methoden erhaltenen
Resultate sind in Einzeltabellen nachfolgeend zusammengestellt.

V. Ergebnisse der Untersuchungen.

Fassen wir die Hauptergebnisse der vorliegenden Arbeit

hier kurz zusammen, so finden wir:

1. Das Klemmen tritt bei jeder Holzart ohne Ausnahme
mehr oder weniger auf.

2. Das Mass der Klemmung ist sehr verschieden, je nach
Holzarten, aber bei ein und derselben Gattung an-
nahernd tibereinstimmend.

3. Das Klemmen ist nicht voriibergehend, sondern halt so
lange an, bis in Folge der Verdunstung das Schwin-
den beginnt und. bis auf diesen Zeitpunkt nimmt

~ das Klemmungsmass allmahlig zu.

4. Die Dauer des Klemmens ist verschieden, je nach
Holzarten, nach der Dicke einer Scheibe, ferner
verschieden bei berindeten und bei entrindeten Schei-
ben. Je dicker eine Scheibe ist, desto langer dauert
das Klemmen an. Die Klemmungsdauer ist grosser
bei berindeten Scheiben als bei unberindeten.

5. Die Grad des Klemmens nimmt vom Mittelpunkt des
Stammes gegen die Rinde hin stetig zu. (Siehe Tab. I-V.)

6. Das Klemmungsmass wird durch die Rinde erheblich
vermindert, namentlich ist der Einfluss der Rinde beim
Splintholz grésser, als bei den anderen Holztheilen.
Das Klemmen wird namlich um so kraftiger behindert,
je dicker der Bau der Rinde ist.

KOIDE: UEBER DAS KLEMMEN DER HOELZER. eAiede

7. Die Dicke einer Scheibe tibt keinen grossen Einfluss auf
die Klemmungsgrosse aus. (Siehe Tab. XIII.-XVII.)

8. Bei losgetrenntem Splintholz (Splintholz ftir sich
untersucht) ist die Klemmung immer grosser, als
beim Splintholz einer vollen Scheibe. Dagegen zeigt
der vom Splint befreite Kern (Kernholz fiir sich)
kein anderes Klemmungsmass als das Kernholz der
vollen Scheibe. (Siehe Tab. XVIII. und XIX.)

g. Das Klemmen in der Linie eines einzigen Sagschnitts
entspricht annahernd dem Gesammtklemmen im ganzen
Umfange, (Siebe Tab. XX.)

10. Das Klemmmass einer und derselben Scheibe wird
durch die Lange des Sageschnitts sehr bedeutend
beeinflusst, indem es um so grosser ist, je langer
die Kluft (vom Umfang der Scheibe gegen den Mittel-
punkt .zu) ist. (Vergleiche Tab. II. zu I. und Tab.
Wo Aut D/A)

Es geht nun aus den Untersuchungsresultaten der im
Kap. II. angegebenen letzten fiinf Baume eine bemerkens-
werthe Thatsache hervor, dass das Klemmmass des Kern-,
Mittel- und Splintholzes sich etwa wie 1; 2; 3 verhalt, so
dass man annehmen konnte, dass das Klemmensmass durch
eine lineare Function von der Lange des Sagschnitts dargestellt
werden kénne. Nennt man nun die Grésse des Klemmens 0
die Lange des Sagschnitts 7 und eine Constante a, so wird
diese Annahme in folgender Form ausgedriickt.

Os

Um nun zu prifen, soweit diese Annahme sich der
Natur anpassen lasst, habe ichmit giitiger Unterstiitzung
von Herrn Prof. Dr. Kitao mittelst der kleinsten Quadrate
folgende Resultate erhalten.

312

KOIDE: UEBER DAS KLEMMEN DER HOELZER,.

I. ACER PALMATUM (Momiji).

a=0,0094.
No. der | Distanz| Gemessene| Berechnete Fehler
Scheibe. cm. cm. cm. cm.

(22 0,12 0,1128 —0,0072
I. | 8 0,07 0,0752 +0,0052
4 0,03 0,0376 -+0,0076
( 12 0,12 0,1128 —0,0072
Iie 1 8 0,06 0,0752 +0,0152
Rew 671 0,03 0,0376 -++0,0076
12 0,12 0,1128 —0,0072
III. | 8 0,07 0,0752 +-0,0052
4 0,04 0,0376 —0,0024
12 0,12 0,1128 —0,0072
IV. 8 0,07 0,0752 -+0,0052
4 0,03 0,0376 +0,0076
12 0,12 0,1128 —0,0072
V. 8 0,07 0,0752 +0,0052
4 0,04 0,0376 —0,0024

Bemerkungen.

Der wahrschein-

lichste Werth
des wahrschein-
lichen Fehlers
f=0,0050

II. HALESIA CORYMBOSUM (Asagara).

No. der | Distanz
Scheibe. cm.
9
1. 6
3
9
te 6
3
( 9
Ill. | 6
3
( 9
IV. | 6
| 3
| 9
i
3

a=0,0102.

Gemessene| Bereclinete Fehler
cm. cm. cm.
0,09 0,0918 +0,0018
0,05 0,0612 +0,0112
0,03 0,0306 +0,0006
0,09 0,0918 +0,0018
0,05 0,0612 +0,0112
0,02 0,0306 +0,0106
0,10 0,0918 —0,0082
0,05 0,0012 +0,0112
0,03 0,0306 +0,0006
0,11 0,0918 —0,0182
0,06 0,0612 +0,0012
0,03 0,0306 +0,0006
0,10 0,0918 —0,0082
0,06 0,0612 +0,0012
0,03 0,0306 +0,0006

Bemerkungen.

f=0,0056

KOIDE: UEBER DAS KLEMMEN DER HOELZER. 313

III. BETULA ULMIFOLIA (Yoguso-minebari).

a@=0,0104.

Bemerkungen.
No. der | Distanz| Gemessene} Berechnete Fehler
Scheibe. cm. cm. cm. cm.

0,09 0,0988 +0,0088
0,06 0,0676 -++0,0076
0,03 0,0312 +0,0012

0,10 0,0988 —0,0012
0,07 0,0676 —0,0024
0,03 0,0312 +0,0012

ONnH OUH OHH

0,10 0,0988 —0,0012
0,06 0,0676 -+0,0076
0,03 0,0312 +0,0012

0,10 0,0988 —0,0012
0,07 0,0676 —0,0024
0,03 0,0312 +0,0012

0,11 0,0988 —0,0112
0,07 0,0676 —0,0024
0,04 0,0312 —0,0088

CouMM OoOuNM

IV. FAGUS JAPONICA (Jnu-buna).

a=0,0126.
Bemerkungen.
No, der | Distanz| Gemessene | Berechnete Fehler
Scheibe. cm. cm. cm. cm.
9 0,12 0,1134 —0,0066
I, | 6 0,08 0,0756 —0,0044 f=0,00341

3 0,04 0,0378 —0,0022

9 0,1 0,1134 +0,0034
II, 6 0,08 0,0756 —0,0044
3 0,04 0,0378 —0,0022
9 0,1 0,1134 +0,0034
Ill. 6 0,08 0,0756 —0,0044
3 0,04 0,0378 —0,0022
9 O,II 0,1134 +0,0034
IV. 6 0,08 0,0756 —0,0044
3 0,03 0,0378 +0,0078
9 0,11 05,1134 +0,0034
We { 6 0,08 0,0756 —0,0044
3 0,03 0,0378 +-0,0078

314 KOIDE : UEBER DAS KLEMMEN DER HOELZER.

V. TSUGA SIEBOLDII (7suga).

a=0,0098.

. Bemerkungen,
No. der | Distanz| Gemessene} Berechnete Fehler
Scheibe. cm. cm. cm. cm.

—o,o118
+0,0088 f=0,00998
+0,0194
—o0,0018
+0,0188
+0,0194
—o0,0118
+0,0088
+0,0194
—o0,0118
—0,0112
+0,0194

—o,o118
—0,0012

+0,0194

WOAOW WAO WAO WAO W AO

Wenn man bei dieser geringen Anzahl der untersuchten
Scheiben zu einem allgemeinen Schluss berechtigt ware, so
kann man sich bei diesem Werth des wahrscheinlichen
Fehlers der Thatsache nicht verschliessen, dass das Klemm-
mass sich in der That mit grosser Annaherung durch eine lineare
Function von der Lange des Sagschnittes darstellen lasst,
da doch die Fehler wie der Vergleich ihrer Werthe mit dem
wahrscheinlichsten Werthe des wahrscheinlichen Fehlers zeigt,
Groéssen haben, die fast ebenso oft tiber wie unter (f) liegen,
also dass man wohl berechtigt ist, die Abweichungen zwischen
dem beobachteten und berechneten Klemmen dem unvermeid-
lichen Beobachtungsfehler zuzuschreiben, der freilich bei der
keine allzu grosse Genauigkeit gestattenden Messungsmethode,
wie bei der Geringfiigigkeit der zu messenden Lange sicher
ein bedeutender sein mochte.

315

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

I. LARIX

LEPTOLEPIS (Kavamatsu).

Volle Scheibe mit Rinde.

Volle Scheibe ohne Rinde.

n
Vv
° 5 =& a ste} o $3 | o
io. Wcetre ers Se |s2 | 2 Se |se | 4
~ a = 20 DD op Bezeichnung oe Pe) 2) a as S)
A Wael ae cialeer feg|eae| & Pe | hee | &
n |e F/B S : i164 o0 o/ : S : o0
5 | 8 |gen8 (ous & der SGG|e28|) 6 | % |SSE| aa 8 55
ao] GH nH os) o & vo
A Q bay ar ne) Holzstiicke. ie g 5 S| SI 5 FS E &
is a2)
g Ba” | ge ge jee | 5 eo a8 | §
a ae v3 wn i 2 n M
y —_
{ Splintholz velista! 4550 4.31 0,19 4,2 4,65 4544 0,21
is I 28,5 16,4 ee ieceenzone Sagal) Sys 3,32 0,15 453 3,58 3,41 0,17
Kern OlZiyeveretelaletelesi| kya S 1,44 0,04 257 1,55 1,49 0,06
Splintholz paocosonl! Skisk 3,68 0,15 3,9 3,92 3,73 0,17
Tile Zwischenzone ....| 2,63 2,52 O,II 4,1 2,02 2,50 0,12
SWAN Sogoonuoud) 1,39 0,03 2,1 1,45 1,40 0,05
Splintholz auauecal Zifey! 3,90 0,14 354 3,99 3,83 0,16
Zwischenzone ....| 2,84 2,73 0,11 18 2,601 2,50 0,11
7 3 5
IennholZverielevcietsrsretel| 70) 1,6 0,0 25 1a rsa 0,06
7 5 5 9 57 5
Splintholz. ........] 3,74 3558 0,16 4,2 an77 3,60 0,17
Zwischenzone ....| 2,50 2,40 0,10 4,0 2,59 2,48 O,II

EennholZisviwrererereais

%

457

BEMERKUNGEN.

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

II. LARIX LEPTOLEPIS

No. der Scheibe.

Baumhohe

m.

Durchschnittlicher

Durchmesser
der Scheibe

cm.

| Lange des Sagschnittes

vom Mark bis zum
Umfang

cm.

Bezeichnung
der

Holzsticke.

Splintholz ...
Zwischenzone
Kernholz .....

Splintholz ....
Zwischenzone
Kernnolzmrieite

Splintholz ...
Zwischenzone
WermhiOlz ers eater

Splintholz ...
Zwischenzone
Kernholz.....

Volle Scheibe mit Rinde.

(Karamatsu).

Sehnenlange vor
dem Schnitte

cm.

Sehnenlange nach

dem Schnitte

Klemmungsgrésse

Volle Scheibe ohne

Rinde.

oS
Sehnenlange vor
dem Schnitte
cm

Sehnenlange nach
dem Schnitte
cm.

Klemmungsgrésse

|

298
onr
-_ Hw CO

go9
oon
Ww op

9
a]
_

BEMERKUNGEN

317

UEBER DAS KLEMMEN DER HOELZER,

KOIDE

III. PINUS DENSIFLORA (Akamaisu).

Volle Scheibe mit Rinde. Volle Scheibe ohne Rinde.
n
o _ f= u cl o & a o
3 a ieee Se 32 3 Se |e 3
3 2 =d8 | 3 aS 2 =r, bas ie
3 a ves] (are DD oy Bezeichnung va =| 5 ve aie Sh
3 i s eee , peo & ; ale i She aie P Se ; be 2s 5
H iS 0 co is 2) 2)
, | 8 |\seaSloueé der SGE|ER8| $8 | © | SRE sa5| 86 %
ee i) Sess Bie ee : ae | oie E Be. ioe 5
; a @) Blas) |S Holzstiicke, ao 0 eI ao ao §
S BA Orn ou aU x) ou gu a)
a) £3 a ep MM ft D MM
3 oS) eee oe c E
(Splintholz souooooull, «zhyu 4,22 0,19 4:3 4,31 4,11 0,20 4,6
I, 2 Ba,3 18,0 Zwischenzone ....| 3,32 3,18 0,14 4,0 3,31 3,17 0,14 4,2
\igemnbole spanoaaoanl tele) 1,60 0,06 3,6 1,70 1,64 0,06 335
; (Splintholz ........| 4,26 4,09 0,17 4,0 4,30 4,10 0,20 4,6
II, 4 30,5 17,0 |\Zwischenzone ....| 3,23 3,10 0,13 4,0 3,20 3,05 0,15 4,6
(Kernholz .......50+ 1,70 1,64 0,06 355 1,72 1,66 0,06 355
SPlimtholZ/y circ eters! e477 4,20 0,17 4,0 4534 4,14 0,20 4,6
Ill. 6 29,0 15,8 |2vischenzone ealetell 9540 3,33 0,13 3,8 3,45 3,29 0,16 4,6
KGSIMeO4 soudoagacall Uy 7/3} 1,67 0,06 35 1,81 1,74 0,07 3,9
SplintholZ "sire. esi 4,25 4,06 0,15 3,6 4,22 4,05 0,17 4,0
IV. 8 27,3 12,9 Zwischenzone ....| 3,13 3,02 0,11 a5 3,29 3,16 0,13 3,9
SOSA So0codo0u0n)| sts) 1,53 0,05 shu 1,59 1,53 0,06 3,7
Splintholz ........| 4,22 4,07 0,15 3,6 4,23 4,06 0,17 4,0
Vv. 10 25,9 12,7 |Zvischenzone Soul. Sip7/ 3,06 0,11 354 3,11 2,98 0,13 4,0
Kernholz...esseeee| 555 1,50 0,05 3,2 1,52 1,46 0,06 3,9

BEMERKUNGEN.

UEBER DAS KLEMMEN DER HOELZER,

KOIDE

318

No. der Scheibe.

II.

1GOG

IV.

Baumhohe
m-

2,3

43

Durchschnittlicher

&
Zé
= 3

58 tej

na On

nO a8 tof)

25 ; ate FE

[s) & 50 Mas
ao5|ORE5

53 |gaP

Ls]

Q VE
Sr}
es
ww”

4

31,5 16,5

20,5 W555

29,0 14,3

27,0 12,7

25,0 13,1

(

(

IV. ABIES UMBELLATA (Uvashivomomi).

Volle Scheibe mit Rinde.

Volle Scheibe ohne Rinde.

wy < o u o
Bezeichnung oS ne 5p os Sz
cea) fee| & ea | 28
aie 5 e ll aes
der SG6|835| 86 | % | 888/838
eee Se |Ge S Se |¢ée
olzstiicke, 68 Es § 68 2a
vp DB M D D
Aeusserer Theil... 4:50 4,32 0,18 4,0 4,51 4,29
Mittler Theil ..... 3,05 2,93 0,12 3,9 3,01 2,86
Innerer Dheil\ 7...) 1758 1,54 0,04 2,5 1,55 1,50
Aeusserer Theil... 4,08 3,92 0,16 4,1 4,08 3,89
Mittler Theil ..... 2,73 2,62 O,I1 4,0 2,75 2,63
Innerer hele. 1,41 1,36 0,05 355 1,45 I,40
Aeusserer Theil... 4,23 4,07 0,16 4,0 4,26 4,08
Mie 400@U Gooac 2,87 2,76 0,11 3,8 2,91 2,79
Innerer Tibet cer. 1,44 1,40 0,04 2,7 1,52 1,47
Aeusserer Theil ....] 4,41 4,26 0,15 354 4:43 4,26
Mittler Theil ..... 3,22 Bee 0,10 Bar 3,23 Shayene
Innerer) Dheil reel 2,00 1,56 0,04 2,5 1,68 1,63
Aeusserer Theil ....} 3,90 3:75 0,15 3,9 4,47 4,29
Mirttlerslherlieerctare 23 S5 2,74 0,11 3,8 3,01 2,89
Inneren Phe... 1,44 1,40 0,04 2,7 1,50 1,51

Klemmungsgrésse
cm.

BEMERKUNGEN

319

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

7

No. der Scheibe.
der Scheibe
cm.
| Lange des Sagschnittes

V. ABIES UMBELLATA

Bezeichnung

der

Umfang
cm.

Holzsticke.

Durchmesser
vom Mark bis zum

Baumhohe
m-
Durchschnittlicher

Sehnenlange vor

dem Schnitte

Volle Scheibe mit Rinde.

cm.

Sehnenlange nach

dem Schnitte
cm

Klemmungsgrosse
cm.

Volle Scheibe ohne Rinde.

(Uvashtromomt).

Sehnenlange vor
dem Schnitte

cm.

Sehnenlange nach

al

dem Schnitte

Aeusserer Theil ..
Mittler Theil ....
Innerer Theil...

(Aeusserer Theil
Mittler Theil ..
Innerer Theil...

Aeusserer Theil
Mittler Theil ..
Innerer Theil ..

Aeusserer Theil
Mittler Theil .,
Innerer Theil ..

Mittler Theil ..

Aeusserer Theil
{itter ANG :

grosse

Klemmungs
cm.

BEMERKUNGEN.

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

320

No. der Scheibe.

Baumhéohe

der Scheibé

m.
Durchschnittlicher
Durchmesser

cm.
Lange des Sagschnittes
vom Mark bis zum
Umfang

cm.

VI. CRYPTOMERIA JAPONICA (Sug7).

Bezeichnung
der

Holzsticke.

Il.

Tie

IV.

10

253

2335

21,5

20,5

19;3

15,3

13,0

12,1

10,7

9,8

Splintholz ....
Zwischenzone
Kernholz......

Zwischenzone
Kernholz......

Splintholz ....
Zwischenzone
WennhiOlZrreireers

(Splintholz ....
Zwischenzone
KernholZementsts

Splintholz ....
Zwischenzone

Kernholz.......

{atwischen eee

Volle Scheibe mit Rinde.

Sehnen!ange vor
dem Schnitte
cm.

4,51
3,58
1,78

4,22

Se
1,64

4517
3,03
1,60

4,07
3,01
1,66

4,01
2,90
1,48

Sehnenlange nach
dem Schnitte
cm.

4,41
3,50
1,76

4,12
3,16
1,62

407
2,96
1,58

3:97
2,94
1,64

3,91
2,88
1,46

Klemmungsgrosse
cm.

ok

2,2
2,2
Tp

2,3

2,4
te)

254
2,3
1,2

2,4

2

2

2,4
2,3
1,3

Volle Scheibe ohne Rinde.

GC vo
Pe) Pw] Lea)
: (S| ; fo)
28/828) $6 | %
o Ss &
a & o§
ov fo &
ov oo x)
a n i
437 | 4:26 | 0,x1 2,5
3,53 3545 0,08 2,2
1,80 1,77 0,03 1,6
4,27 4,16 0,1 2,5
3,21 3,13 0,08 2,5
1,49 1,47 0,02 2,0
4,09 3,98 O,Ir 2,6
3,10 3,02 0,08 2,5
1,48 1,46 0,02 2,0
4,07 3,96 0,11 2,6
3,06 2,98 0,08 2,6
1,53 1,51 0,02 1,9
3,95 3,84 0,11 2,6
2,92 2,85 0,07 254
1,17 1,67 0,03 1,6

BEMERKUNGEN,

321

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

VII. CHAMAECYPARIS OBTUSA (Hinok:).

Volle Scheibe mit Rinde.

Volle Scheibe ohne Rinde.

No. der Scheibe.

Baumhohe

m.

Durchschnittlicher

Durchmesser

der Scheibe

cm.

Lange des Sagschnittes

vom Mark bis zum

Bezeichnung

der

Umfang
cm.

Holzsticke.

Splintholz ....
Zwischenzone
Kernholz ......

Splintholz ....
Zwischenzone
ermholZiererctere

Zwischenzone
KernholZieterelete

Splintholz ....
Zwischenzone
INernhnolzpesisere

Splintholz ....
Zwischenzone
Kernholzi.s ..00

eeee

seer

seer

we a re Lal a a
Se |g2 ] 8 sg |e2 | 3
o's ors Sp v= oe op
oo & 50 & a bo S bo & n
go eas ons ) BSe|/ ese on
snG|sa8) 26) 2 |Sa8|sa8) 88
Se |e E 92 |e :
fg | os 5 Su | as o
n Dn M na n M
4554 4543 0,11 2,4 4,60 4,48 0,12
3555 3575 0,10 2,5 3,80 | 3,80 | 0,10
2,07 2,04 0,03 I,4 1,80 1,77 0,03
4,56 4545 O,Ir 2,4 4,57 4,45 0,12
3595 3,86 | 0,09 2, 3193 3,83 | 0,10
215 aye 0,03 I,4 1,87 1,84 0,03
451 4:40 | 0,11 2,4 | 4:59 | 4.47 | 012
3,80 3,72 0,08 2,1 3,82 3,73 0,09
2,02 2,90 0,02 I,0 2,10 2,07 0,02
4544 4534 0,10 2,3 4543 4532 0,1T
3573 3,65 0,08 2,1 3,66 3557 0,09
1,80 1,78 0,02 at 1,97 1,95 0,02
4:57 | 4:46 | 0,tr 2:4 | 4:34 | 4:23 | O,t2
3575 3,66 | 0,09 2,4 | 3,79 | 3,62 0,09
1,92 1,89 0,03 1,5 1,88 1,85 0,03

BEMERKUNGEN

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

322

Lange des Sagschnittes

vom Mark bis zum
Umfang

cm.

Bezeichnung
der

Holzstiicke.

; 5D

a Su

me o 03
o an
£ a= zat ie
3) i 26.9
n E¢ Seog
uw CN o
o 3 Cy)

Eo} GH Qew

Q apes

5 Oobdwd
°o =e)

a A

15 I 23,0
1) 2 22,0
III, 3 22,0

10,6

12,2

10,8

Aeusserer Theil ..
Mittler Thell ....
Innerer Theil......

Aeusserer Theil ..
Mittler Theil .....
Innerer Theil.....

Aeusserer Theil..
Mittler Theil ....
Innerer Theil.,..

Aeusserer Theil..
Mittler Theil ....
Innerer Theil ....

BETULA

Aeusserer Theil...
Mittler Theil .....
Innerer Theil ....

ALBA, var VULGARIS (Siz

Volle Scheibe mit Rinde.

Sehnenlange vor
dem Schnitte
cm,

cm.

Sehnenlange nach
dem Schnitte

Klemmungsgrésse
cm.

Q

1,3

Sehnenlange vor
dem Schnitte
cm.

vakaba)

Sehnenlange nach
dem Sch itte
cm.

Klemmungsgrésse
cm.

Volle Scheibe ohne Rinde.

BEMERKUNGEN.

323

IX. MAGNOLIA HYPOLEUCA (Hohonok:).

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

Volle Scheibe mit Rinde. Volle Scheibe ohne Rinde.
n
Ls 3 & a o <— o
o v B=) hw rs} D S O 5
3 r= 3 52 é 5 Bezeichnung 5 # ES : 4 £ E = °
Poa 288 | See ‘ ee | Ss .| @. , |eeal Peel be
=} q ] iS iy ia O fs] o/ | 1 i fo)
. gs SER 5 owe § der Sas id § a ifs) aa aa § a 7%
seh) SI Eppa RCPS =) ; ee | sé E ee | es 5
3 2qQau og Holzstiicke. a8 Ea g 2 a 8
a A Bie a a M HB B x
iw >
- Ec a
SplintholZeeerrenreelele| oi 4,31 0,00 0,0 4,19 4,16 0,03 0,7
i, 0,3 32,0 12,5 [Zvischenzone all Se) 3,29 0,00 0,0 3533 3,31 0,02 0,6
OPV Gocagbadad) 24s 1,78 0,00 0,0 1,76 1,76 0,00 0,0
Splintholz ........| 4,05 4,05 0,00 0,0 4,15 4,12 0,03 0,7
II. 253 27,5 12,2 {Zwischenzone soda Shor 3,04. 0,00 0,0 2,10 3,08 0,02 0,6
LINO sacogacovnl) 2/85} 1,43 0,00 0,0 1,50 1,49 0,01 0,7
(Splintholz Goooumonl Sick 4,88 0,00 0,0 3,09 3,93 0,06 1,5
III. 4;3 24,5 10,8 Zwischenzone ....| 2,93 3,93 0,00 0,0 2,94 2,90 0,04 1,4
RIN Saocoonadell sy 1,57 0,00 0,0 1,50 1,48 0,02 1,3
Splintholz ........| 4,07 4,05 0,02 055 4,1 4,05 0,06 1,5
IV. 6,3 22,8 954 [Zivischenzone Agoal! Sk 3,04 0,01 0,3 3,19 3,14 0,05 nS
NEXELmOlZisinielelele
Splintholz ....
Zwischenzone
INGTINOl Zia tereteretavereiale

BEMERKUNGEN.

X. PRUNUS CERASOIDES (Mejirosakura).

UEBER DAS KLEMMEN DER HOELZER.

.

KOIDE

324

Volle Scheibe mit Rinde. Volle Scheibe ohne Rinde.
5 3 Ze . 5 2 * 4 2 c)
2 She |88 Se |ay ig S2 |e Gy A
a a 34a Ou Bezeichnung ow cs ae ae aS se) 5
a: 3 52°59 nse 00 Ei oe bp ore v's S09 iw
® | ea |SEpelaude esa/P5e| Se | % |eee|#ee| Be| x | 2%
ie EE OENO|V REO dee eno san § 55 7 eNo|8NG 50 % ica
a ee a een teers ei eecedl fe |Ge | & ce | Ge | 8 a
5 Bay vs olzstucke, ag ae e 68 as 5 a2}
Z Qa & 2 Nn Be) ) wa n 17,
— —— y — —

| Splintholz so0c0R ON] Zhyte 4,34 0,04 0,9 4,22 4,17 0,05 1,2

I. 0,3 22,5 6,7 beeeenzone eve] 3547 3344 0,03 0,9 3344 3,40 0,04 Tepe

STOW, sa acoqooca) {5 1,63 0,02 53 1,99 1,97 0,02 1,0

(Splintholz Soooonod) 2st) 4134 0,05 iigut 4,26 4,20 0,06 1,4

I; 2,3 20,5 7,8 )Zwischenzone ....| 3,54 4,50 0,04 Mepe 3554 3,49 0,05 I,4

(Kernholz eelejsivle\eiolse]| LZ 1,68 0,02 Tar 1,70 1,68 0,02 I,I

(Splintholz Scan0000)- Zs 4,28 0,06 1,3 4,33 4,26 0,07 1,6

NOE 453 19,3 6,9 Ie wiseheazone eoes| 3,47 3543 0,04 1st 3,54 3,49 0,05 1,4

KPNI so Ganooasnl) risk) 1,78 0,02 ify) 1,75 1,73 0,02 Lagat

(Splintholz. SApoaaool “ey 4,28 0,06 13} 3,95 3,89 0,06 1,5

IW 6,3 18,3 8,1 ;Zwischenzone ....| 3,55 3,50 0,05 1,4 3,23 3,18 0,05 1,5

Vvernholziems\acetre | imniyo 1,77 0,02 I,I 1,70 1,68 | 0,02 re

DER HOELZER. 325

UEBER DAS KLEMMEN

KOIDE

No. der Scheibe,

Baumhohe

m,

Durchschnittlicher

Durchmesser

der Scheibe

cm.

Lange des Sagschnittes
vom Mark bis zum
Umfang
cm.

XI. ZELKOWA KEAKT (Keak:).

Volle Scheibe mit Rinde.

Volle Scheibe ohne Rinde.

Bezeichnung
der

Holzstiicke.

Splintholz
Zwischenzone ....
Kern holiziererereistatelarels

(Splintholz cclececes
Zwischenzone ....
Krenn OlZierieterstetstelele

Zwischenzone ....

{aswisehen
ISGrmil1 Ol Zmeretetetetepeterere

Zwischenzone ....
WWwernholZeretetererereretels

Splintholz
Zwischenzone ....
Kernholz ...... 000

{Ziishen soadooGn

Sehnenlange vor
dem Schnitte
cm.

Sehnenlange nach

dem Schnitte

Klemmungsgrosse
ro

Sehnenlange vor
dem Schnitte

Sehnenlange nach

dem Schnitte

cm.

Klemmungsgrosse

BEMERKUNGEN.

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

326

XII. JUGLANS SIEBOLDIANA (Onitkurumz).

Volle Scheibe mit Rinde. Volle Scheibe ohne Rinde.
g fa
: 5 Zé x a) o o oO
3B Gl w & = Sa g Oo a S oO a o 7 Z
ome, o = © Y <a 5 ? 3 cv :0 =) ow Fe) 5
| S = 92 on Bezeichnung (a= 2 by os a8 by
5 S £2'5 ces (te oS =| 80 bo o's 8p
® | e¢ Segal sud Bed|fe¢a| Se | o |asg| Pea] be be
we Ee SER OHSS der —nNO6 WN GS Ets % SH58 WO GO g& ve Q
2 S ao. os oa Eg fe og Eg & =
. ma OBS aS) Holzsticke. £ 50 & ao 20 FSI &
S y Q ep € 7] a s mo} o ov aU o [ee]
fa) Es ae a Mm % a S|
=|

SplimtholZyeeiseieiste +4400 4,66 0,03 0,6 4575 4,70 0,05 1,0

Mn 0,3 28,5 12,6 |Zvischenzone slelelal MARZO) 4,24 0,02 055 4,25 4,22 0,03 0,7

IXernWOliZeeiatetsieleleriate eet eO 2,28 0,00 0,0 2,14 2,13 0,01 0,4

SplintholZz) <\j0.. 1 4573 4,70 0,03 0,6 4,72 4,67 0,05 I,1I

iD, 2,3 24,8 10,5 Zwischenzone ....| 4,24 4,22 0,02 0,5 4,28 4,24 0,04 0,9

ISenmhOlZeretelaiststelatels|| 2s 0.5 2,05 0,00 0,0 222, 2,21 0,01 0,4

(Splintholz sandsoon| Zs 4,81 0,04 0,8 4574 4,68 0,06 I,2

Ill. 4,3 24,0 9,8 (eweceenzone codGl| Zlnshs! 4,35 0,03 0,7 4,30 4,26 0,04 0,9

CANN! ooononc000| ZZ} 2,22 0,01 0,4 1,99 1,98 0,01 0,5

Splintholz ........| 4,78 4,73 0,05 1,0 4,70 4,63 0,07 1,4

IV. 6,3 22,5 9,0 Zwischenzone ....| 4,38 4,34 O O4 0,9 4531 4,26 0,05 righ

OMNI sodaoacoanl Ay27/ 2,26 0,01 0,4 2,25 2,24 0,01 0,4

DER HOELZER. 327

UEBER DAS KLEMMEN

KOIDE

XIII. FAGUS JAPONICA (Jnubuna).

Volle Scheibe mit Rinde.

in Im

No. der Scheibe

Hohe.

Dm. in 1m Hohe
cm.

24

24

24

24

Z |
fea} }
“4 7) 1)
ne 1 OD sO WW G o
lasts s2 Se @y 2 iS
5 a ig as Bezeichnung der Or aoe So M
HO: (8852) oes Bee SE oo
fh icone alleen 0g GOs Ss o%, a
oO von? ee i eAo SN SO 30 = }
iS OS a4 Holzsticke. ae BE iS s)
O WE y Sa = oO co & foc] |
= Woes 20 ov cH uv a) H
i
9 AVELISSErer DNerlcsrcel: 3,00 2,88 0,12 4,0 |
I II 6 Mittlersivhenleercerelsre 3,00 2,92 0,08 2,6 i
3 Innere hele src 3,00 2,96 0,04 153
9 Aeusserer Theil... ... 3,00 2,89 0,11 3,6
2 II 6 Minttlersilverleprercteerete 3,00 2,92 0,08 2,6
3 Innerer helices 3-00 2,96 0,04 1,3
9 Aeusserer Theil...... 3,00 2,89 0,11 3,6
3 II 6 Mittleriiierlircreisjelere 3,00 2,92 0,08 2,6
3 Moree Weyl Goooan 3,00 2,96 0,04 1,3
( 9 Aeusserer Theil...... 3,00 2,89 O,II 3,6
4 II 6 Mittler Dheiliiecc. cl. 3,00 2,93 0,08 2,6
\ 3 Mnnerers Dbene ea. 3,00 2,07 0,03 1,0
( 9 Aeusserer Theil...... 3,90 2,89 0,11 3,6 :
5 Ir {© IMittlerD hetliveysjeree are 3,00 2,92 0,08 2,6
( 3 Tnnererajne merece. 3,00 2,07 0,03 I,0

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

No. der Scheibe
in 1 m Hohe,

bis zum Umfang

Lange des Sag-
schnitts vom Mark

cm.

XIV. ACER PALMASUM (Momiji).

Distanz des Marks
von Messpunkte
cm.

Volle Scheibe mit Rinde.

Bezeichnung der

Holzsticke.

Ill.

IV.

oO oO
2 2
a 5

d (dp) 6
Se eee

oO ovo
t= 15}
&
iB A
30,2 I
30,2 2
30,2 3
30,2 4
30,2 5

13,4

13,4

13,4

Ae, oO —_—— —-—_*

Aeusserer Theil..

While Winetlesas aos

Innerer Theil ..

Aeusserer Theil..
Mittler Theil....
Innerer Theil ..

Aeusserer Theil. .
Mittler Theil....
Innerer Theil ..

Aeusserer Theil...
Mittler Theil....
Innerer Theil ..

Aeusserer Theil. .
Mittler Theil....
Innerer Theil ..

Sehnenlange vor
dem Schnitte
cm.

st cS)
S a
as 9
o's 50
Opte One
Goes op
iu. is) 5 E
o§ 8
ES 5
D M
2,88 0,12
2,93 0,07
2,07 0,03
2,88 0,12
2,04 0,06
2,97 0,03
2,88 0,12
2,03 0,07
2,96 0,04
2,88 0,12
2,93 0,07
2,97 0,03
2,88 0,12
2,93 0,07
3,96 0,04

2,3

BEMERKUNGEN.

29

3

UEBER DAS KLEMMEN DER HOELZER.

KOIDE

XV. BETURA ULMIFOLIA (Yoguso-minebari).

Volle Scheibe mit Rinde.

Bezeichnung der

Holzsticke.

No. der Scheibe
in 1 m Hohe.
Dm. in 1m Héhe
cm.

Dick der Scheibe
cm.

Lange des Sag-
schnitte vom Mark
bis zum Umfang
cm,
Distanz des Marks
von Messpunkte
cm,
Sehnenlange vor
dem Schnitte
Sehnenlinge nach
dem Schnitte
cm.
Klemmungsgrosse

BEMERKUNGEN.

Splintholz ....
Zwischenzone
ING SnolZuererete

Hw

Cnn

Splintholz ....
Zwischenzone
Kernel Zameen

Wao Ww ano
Hpw

Splintholz ....
Zwischenzone
Kernholz)s...

DO
ouw oun

HNnw

Splintholz ....
Zwischenzone
Kernholzy "nv

CAN

pw

BH COW OBWW OOD

Splintholz ....
Zwischenzone .
errilialziem cnc cisleiches

YPHO wand Ww

Onn

UEBER DAS KLEMMEN DER HOELZER.

.
.

KOIDE

XVI. HALESIA CORYMBOSUM (Asagara).

Volle Scheibe mit Rinde.

“4
o o i

eho ea 5 ws ge
og 36 a nas
One 2) aé& &
nr i=l Oe OO)
ee ra 5 o 5 ie. g 5
3 | mS fox 3
og a5 6 fee ey
Ze rat a Bie

i 21,4 I 11,5
II. 21,4 2 11,5
III. 21,4 3 11,5
IV. 21,4 4 11,5

V. 21,4 5 11,5

_—— CSS == == a

Distanz des Marks

von Messpunkte

WOO WDOoO WAO WAO W DO

cm,

Bezeichnung der

Holzsticke.

SPlinthOlZ welete)<-ieielels
Zwischenzone ......
Kernholz

SPlinthOlZMerereistetelelelele
Zwischenzone ......
Kernholz

SplintholZrreretelelelele's
Zwischenzone .....-
Kernholz

Splintholz ..........
Zwischenzone ......
Kernholz

eeeeseesee

Splintholz ....cceees
ZwischenZOne ..vece
Kernholz

Sehnenlange vor
dem Schnitte
cm.

§ 8
oO n
BS 2
voc
NOG « &p fe
aa § a8
& E
oa §&
23 §
D M
2,91 0,09
2,95 0,05
2,97 0,03
2,91 0,09
2,95 0,05
2,98 0,02
2,90 0,10
2,95 0,05
2,07 0,03
2.89 O,II
2,94 0,06
2,97 0,03
2,90 0,10
2,04 0,06
2,07 0,03

BEMERKUNGEN,

33

DER HOELZER.

UEBER DAS KLEMMEN

.
.

KOIDE

XVII. TSUGA SIEBOLDII (7suga).

Volle Scheibe mit Rinde.

BEMERKUNGEN.

o © o “ rs o0 a vo Lal a “
25 | 3 3 |#s3 Ea Sy go a
eS o 3 Wee a5 Bezeichnung der One oe So
2 n§ na, ee a0 D

ne Es nN. Sy Go cos s OD

& wf |oS el] gag So & & @ (2 ie

at fs Lal) go bh eo Seo Eno LSTA (3) 50
ne) ia 45 SG bo 5 aa Holzstiicke. ore 5 fe &
og E Oo aces Sie nae gos &
Tie a olesiga G2) a oO =
Q 9A fay Se n mM

9 Aeusserer Theil...... 3,00 2,90 0,10

Te 25,4 I II,2 6 Mittler Theil........ 3,00 2,95 0,05

3 Morne Wel coocac 3,00 2,99 0,01

9 ANENISSEKer eh etlsererenye 3,00 2,91 0,09

10K 25,4 2 II,2 | 6 Mittler Theil 5066 : 3,00 2,96 0,04

3 Innerersdilrerlierecrciers 3,00 2,99 0,0L

9 Aeusserer Theil...... 3,00 2,90 0,10

Te 25,4 3 II,2 6 Mittler Theil.... 6 3,00 2,95 0,05

3 Innerer Theil .. 3 3,00 2,99 0,01

9 NGUSSELE TDCI rrerersr 3,00 2,90 0,10

IV. 2554 4 11,2 | 6 Mittler Theil.. eretaterets 3,00 2,93 0,07

3 Innerer Dheil) ~~... 3-00 2,99 0,01

9 Aeusserer Theil...... 3,00 2,90 0,10

V. 25,4 5 II,2 6 Mittler Theil........ 3,00 2,04 0,06

3 InnererpD heli ayetersres 3,00 2,99 0,01

or
wa

Wow wWHw

SoPw ONnW

332 KOIDE: UEBER DAS KLEMMEN DER HOELZER.

_— .
= a No. der Scheibe.
5 tad i
Baumhohe
Ww iS) cal m.
S 9 . Durchschnittliche
ae Fa ot Durchmesse
cm.
Sehnittlange des
oe EE SS kerns
z: o a cm.
ae ni Breite des Splintes
bh Rey be cm.
w us wo Sehnenlange
° re) 8 vor dem Schnitte
2 ) cm.
» S rs Sehnenlange nach n
a) No) © dem Schnitte Aes
v . © cm. 5
eH
io) =]
& & © Klemmungsgrésse £78
(e) fo) H 5S =) eH
co lo) ° cm. eS es
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ie)
>
s N w 38 3
fo) [o)) Les) =:
= re r
S na ss Sehnenlange nach te)
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=
i moa
& & & Klemmungsgrosse aes
ro] 4 H 5 SB
) H - cm. e oa)
n
z.
on
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N “NI N cm. @,
o
o
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& g & Klemmungsgrésse =O
° ° ° Be
w w Ww cm. Po
oO
Las!
5
a
Lal Cal cal aw h
° re} ° 9 =

BEMERKUNGEN.

‘TIITAX

“(angauiut-osnsoX) YWITOAINTA VIOLAL

KOIDE: UEBER DAS KLEMMEN DER HOELZER.-

No. der Scheibe.

Baumhohe
ow n ro) m.

Durchschnittliche

is} iS} nN
3 2 & Durchmesser
= cm.
Sehnittlange des
& Sy & kerns
[o)) Lal al cm.
x s w Breite des Splintes bd
N fo) [o-) cm, eH
brs @ we Sehnenlange
°° To) t vor dem Schnitte
° ° fo) em. a
3 re Sehnenlange nach yn i.
o ‘oo ve) dem Schnitte ack fe3|
© ne 2 cm. St ™M
sh >
3
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Bh |) od
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= » Sehnenlange nach 2 fan)
re) ‘bo oS dem Schnitte = mM
© © S cm, 5 q
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cal co 3c: -_—™
ra) 2) fe) cm. an mh
om eR
2)
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op oe iy re g. =
w [o) w 5 8
o 7
S g » Sehnittlange nach D
io © re) dem Schnitte @
“I N (oy) cm. a.
o
2 2 2 Klemmnngsgrésse a, oe
le) ° ° aa
w w + cm. 5
w
= o
a!
3
° a
Se & a eH
° ° & oO E

BEMERKUNGEN.

333

334 KOIDE: UEBER DAS KLEMMEN DER HOELZER.
_
= a 5s = 3 an}
= » 0g ° r=} 5
og 0Q. c 3 Co
» rs) a = e Cc
. Dp g = x N
: . = . : >
5 ° 3 . = 2)
: : aes : <
5 : S 5 : res]
5 : Zs : 5 Z
w w b < x Durchschnittliche
= 3 = = = Durchmesser
~
4 cm.
w w uw a6 w Sehnenlange
° ° ° °° ° vor dem Schnitte
° ° o ° ° cm
s S ~ 3 e Sehnenlange nach S
re) re) Oo S re dem Schnitte ws
. as ec a 4 cm. we
Bg?
2 & 2 & g Klemmungsgrésse oa
° fo) ° [e} H oe
w > ui fo) ° cm. 3
A bs is és & Sehnenlange nach
° re) ° ° ro) dem Schnitte
° _ 4 ° ° cm.
Z
. 3 ° KI ;
— £ 2 2 & emmungsgrosse
° ° ° ° ° 888 bd
°o _ 4 ° ° cm. bd
Ls
he = . Ae -< Sehnenlange nach
re) o re) re) Ne) dem Schnitte
io} ao io) a o cm.
i) i) i) 2 2° Klemmungsgroésse
° ° ° ° ° an
° Nn - n x .
Ss ~ iS S Sehnenlange nach
re) re) re) re) oO dem Schnitte
ve) © ao “N uw cm.
u
2 2 & © 2 Klemmungsgroésse
° ro) ° ° ° oa
= La NS Ww uw .
a “3 S is 3 Sehnenlange nach
re) re) re) xe) io dem Schnitte
co re) a) fe) a —
<
2 2 2 2 2 Klemmungsgroésse
°
8 2 8 8 a cm.

BEMERKUNGEN.,

Prk etimteeaneZ
RP pk te te ime ecm we

memeM ce 86k ool CU

E =£ «x < (Me Series

RIRE BRM SE Ilb ores

& £ & < @HEew kK =
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| i Ma
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pan .
pice z
\& es ath s
pee we ye me:

Lit

Ertragstafel und Zuwachsgesetz

far
Sugi (Cryptomeria japonica).
Zum Gebrauch fur die japanischen Forstmanner,
VON

S. Honda, Ringakushi et Dr. Oec. publ.

A. O. Professor fiir Forstwissenschaft an der Kaiserlichen Universitat zu Tokyo.

VORWORT.

Die Ertragstafel bildet fiir die Forstwirtschaft dasselbe, was
der Compass fiir die Schifffahrt, ein Instrument, ohne das ein
zielbewusstes Handeln undenkbar ist, und einen ‘‘ Betriebs-
plan” erst erméglicht. Wahrend man nun in Europa, Dank
der Arbeiten der forstlichen Versuchsstationen, zahlreiche Er-
tragstafeln besitzt, fehlen solche bis jetzt noch in Japan mit
seinen eigenartigen Waldern und anders beschaffenen Zuwachs-
verhaltnissen, ein Mangel, der sich in praktischer wie wissen-
schaftlicher Hinsicht in héchstem Grade fiihlbar machte. Diese
Liicke beabsichtigte ich seit langerer Zeit auszufiillen, da sie
mir um so fiihlbarer geworden war, als ich die Waldung des
Kiyosum als Schulwaldung zu bewirthschaften habe.

In den Winterferien des letzten Jahres fand ich endlich
Gelegenheit, als ich mit meinen Schiilern, 24 an der Zahl, eine
drei Wochen dauernde Excursion in jenem Waldgebiet unter-
nahm, die Zuwachsverhaltnisse der Cryptomeria japonica naher
zu studiren, und so eine Ertragstafel fiir diese wichtigste der
japanischen Nutzholzer aufzustellen.

In den beifolgenden Tafeln sind die Resultate dieser Arbeit
niedergelegt, welche freilich durchaus noch als unvollstandig
angesehen werden muss. Indessen bin ich eifrigst damit be-
schaftigt, weiteres Material zu sammeln und die Ertragstafeln so
zu gestalten, dass sie wohl allen Anforderungen geniigen diirften.

Im zweiten Teile dieser Arbeit habe ich das Zuwachsgesetz
und- gang der Sugibestande in Kzyosunu mit denen der deutschen

/7 20%

330 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Holzarten verglichen, da eine solche Vergleichung in Japan
noch nicht ausgefiihrt worden ist-

Es sei schliesslich bemerkt, dass diese Abhandlung haupt-
sachlich fiir die japanischen Forstleute bestimmt ist, welche zum
grossten Theile der deutschen Sprache machtig sind. Ich hoffe,
dass sie durch dieselbe veranlasst werden, auch ihrerseits Zu-
wachsverhaltnisse anderer Nutzholzer unter anderen Ortlichen
Verhaltnissen naher zu studiren, und so zu Aufstellungen von
Ertragstafeln fiir die japanischen Nutzwalder beizutragen.

io ETE.
Ertragstafeln fur Sugi.

I. Hinleitung,

Fiir cine geordnete Forstwirthschaft ist die Herstellung von
Ertragstafeln die erste Bedingung. Diese Tafeln sind die quan-
titative Darstellung des Wachsthumsganges normal entwickelter
und bestockter Bestande fiir verschiedene Holzarten, Standorte
und Betriebsformen. Sie geben Aufschluss entweder lediglich
iiber die Holzmassen- und Zuwachsgrossen, oder auch weiter
iiber Bestandeshohe, Stammzahl und Stammgrundflache u. s. f.
pro Hectar fiir die Bestandesentwicklung in verschiedenem Alter.

Es giebt mehrere Wege, Ertragstafeln herzustellen. Doch
scheinen mir nur folgende drei Methoden wissenschaftliche
Beriicksichtigung zu verdienen :

1. Die erste nahliegende Methode ist, bei einem Bestande
von ganz jungem Alter, wiederholt, entweder jahrlich oder
periodisch eine Bestandesaufnahme vorzunehmen und sammt-
liche wichtige Factoren bis zu dessen Haubarkeitsalter fort-
gesetzt zu beachten. Insbesondere wird der Einfluss verschie-
dener Erziehungs- und Betriebsweisen auf den Zuwachsgang
der Bestande nur durch solche genaue Controlle mehrerer ver-
schieden behandelter Bestaénde ceteris paribus mit Sicherheit
beobachtet werden konnen, und diese Methode ist daher auch
in Deutschland bei der Zuwachsermittlung durch Einfiihrung
bestimmter Versuchsflachen allgemein acceptirt,

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 337

2. Um den sonst allzulangen Zeitraum abzukiirzen, kann
man auch mehrere Bestaénde verschiedenen Alters gleichzeitig
in Beobachtung ziehen. Man erhalt so einzelne Stticke jener
Kurve, welche die Holzmassenzunahme eines Bestandes wahrend
der ganzen Umtriebszeit darstellen wiirde, welche Stiicke viel-
leicht oft nicht genau an einander passen, aber doch, wenn z. B.
Bestande von je ungefahr 20 jaihrigen Altersabstufungen gewahlt
wurden, nach Verlauf von 20 Jahren erméglichen wiirden, die
Holzmassen- bezw. Zuwachscurven genauer festzustellen, als
dies nach der bisherigen Methode moéglich war. Dabei ist es
zweckmassig, mehrere Besténde von derselben Alterstufe zu
beobachten, um etwaige Verschiedenheiten auszugleichen.

3. Durch diese beiden Verfahren wird der Zuwachs der
Bestande direkt ermittelt; man kann aber auch aus der ein-
maligen Aufnahme mehrerer Bestande verschiedenen Alters eine
Reihe der Bestandesmassen fiir alle Alterstufen erhalten und
so den Gang der Massenzunahme ableiten.

Ich habe bei meinen Beobachtungen die letzte Methode
gewahlt, um in moglichst kurzer Zeit zu Resultaten zu gelangen.

Hinsichtlich der Literatur wurden hauptsachlich folgende
Werke benutzt :

Holzmesskunde, von Prof. Dr. Guttenberg in Lory’s Hand-
buch der Forstwissenschaft.

Holzmesskunde, von Prof, Dr. Baur.

Die Fichte, von Prof. Dr. Baur.

Ertragstafeln fiir die Kiefer, von Prof. Dr. Weise.

Holzzuwachslehre in Forsteinrichtung, von Prof. Dr. Weber.

II. Allgemeine Beschreibung der Standorts- und
Bewirthschaftungsverhaltnisse.

Der Kiyosumwald, in welchem wir diese Untersuchungen
anstellten, liegt in Mitteljapan, ungefahr 70 Kilometer siid-
6stlich von Tokyo entfernt, an den Ausliufern des Grenzgebirges
zwischen den Provinzen Awa und Kadsusa bei 35° 8’ N. B.
und 140° 10’ Oe. L. von Greenwich. Hydrographisch bildet jener
Theil die Wasserscheide zwischen “ Amatsugawa” und ‘“ Obi-
tsugawa.” Die grosste Anhohe der Gegend ist der 382 meter

338 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

hohe Myokdsan, welcher rings von Kzyosumiwald umgeben ist
und von dem aus viele kleine H6henziige nach allen Himmels-
richtungen auslaufen ; diese schliessen vielfach mit ihren steilen
Wanden tiefe Thaler ein, welche meist mit Wald bedeckt sind.

Der Kiyosumiwald misst von Siiden nach Norden circa 3,2
und von Westen nach Osten circa 2,6 Kilometer. Der Wald
grenzt im Westen an den 1700 ha grossen “ Staatswald zu
Okusan,” nordlich an den 4600 ha grossen ‘‘Staatswald zu
Tsutsumori,’ gegen Siiden und Osten wird er vom stillen
Ocean bespiilt. Bisher gehérte jener Kzyoswmiwald als Staats-
wald zum Forstamt Ofaki, noch friiher aber zum Klostergute
Seichojt. Seit einem Jahre ist derselbe in einer Ausdehnung von
330,4 ha als Schulwald der Kaiserlichen Universitat Tokyo
zugewiesen worden und dient dem forstlichen Studium und
Experimenten. Andererseits soll er auch ein Muster moderner
systematischer Forstwirtschaft reprasentiren.

Der Untergrund besteht aus Tuff und tertidrem Sandstein.
Der Boden ist im Allgemeinen sehr durchlassig, allein der mit
Wald bedeckte Theil weist eine reiche circa 10 cm. tiefe
Humusschichte auf, welche den Boden frisch erhalt.

Der hochste Punkt der oberen Grenze liegt bei circa 350 m.;
der unterste bei circa 50 m. tiber der Meeresflache.

Das Localklima ist sehr mild. Die durchschnittliche
Jahrestemperatur betragt circa 15,5°, die niedrigste T’emperatur
circa 2°, die héchste 32° C; die Luft ist sehr feucht, wegen des
Stidwindes, der tiber den warmen Kuroshiwostrom herauf-
streicht. Die Jahresregenmenge betragt ungefahr 2000 mm.
Der meiste Regen fallt im April, Juni und Juli; Schnee fallt
selten, oft einen ganzen Winter hindurch gar nicht.

In friiherer Zeit soll in dieser Gegend ein gemischter Wald
von immergriinen Eichenarten (Quercus acuta, Quercus glauca,
Quercus cuspidata u. s. w.), Tannen und Thugen vorherschend
gewesen sein. Da die Benutzung der genannten Holzarten,
wegen schwieriger Abfuhr fast ganz unterblieb, so musste nach
und nach mit der ktinstlichen Pflanzung des mehr werthvollen
Sugt vorgegangen werden, so dass jetzt der vorherrschende
Sugibestand nur mit einigen Tannen untermischt ist.

Die gegenwartig angenommene Umtriebszeit ist jene von
100 Jahren. Die Betriebsart ist ein Hochwaldbetrieb wobei

HONDA ! ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 339

der Kahlschlagbetrieb mit kiinstlicher Pflanzung die Regel
bildet.

Die jetzt vorhandenen Swgibestande sind ganz durch kiinst-
liche Pflanzung entstanden; Die Einsetzlinge waren 3 Jahre
alte 2 mal verschulte circa halb Meter lange Pflanzen. Die
Pflanzenzahl pro ha betrug dabei 6000 im Reihenverbande.
Nach der Pflanzung haute man jahrlich cin oder zweimal das
ganze Unkraut ab; aber vom 5ten oder 7ten Jahre nach der
Pflanzung an kamen keine Arbeiter mehr in den Wald bis nach
der Haubarzeit. Nach altem Gebrauch der dortigen Gegend
wurde nicht durchforstet, starkere Baume wachsen iiusserst
schnell, den schwacheren nur eine kargliche Existenz gewahrend.
Die Holzbestande haben jetzt meist 10-100 jahriges Alter.

III. Darlegung des bei den Bestandesuntersuchungen
beobachteten Verfahrens.

A. Auswahl und Aufnahme der Probeflachen.

In moglichst gleichmdssiger Verteilung durch alle Alter
wurde eine moglichst grosse Anzahl normal bestockter und in
jeder Beziehung geeignet scheinender Probetlachen von minde-
stens + ha Flachengrésse ausgewahlt und zwar Bestande der
besten und schlechtesten Standortgiite. Dieses gelang uns bei
der provisorischen Bonitirung nach dem Augenmaase_ unter
Zuhiilfenahme der HOhen. Sogern man auch alle Probeflachen
einen Hectar gross genommen hatte, so war dieses doch nicht
moglich, weil die Bestande selten sind, in welchen man in allen
Teilen ganz normal bestockte Flachen von zr ha Fiachen-
inhalt finden konnte.

Unser Plan war ferner, zuerst von 10 zu 10 bis 100 Jahren
je 8 Probeflachen und zwar je vier fiir die besten Bonitaten als
auch fiir die schlechtesten auszuwahlen. Da das nicht voll-
standig modglich war, weil wir in dem dortigen Waldgebiete
nicht gentigende passende Besténde vorfanden, so mussten wir
mit nur 56 Probeflachen, statt 80, einstweilen zufrieden sein,
und schliesslich 15 Probeflachen aus cinem Privatwalde in der
Umgebung unserer Schulwaldung zu Hiilfe nehmen.

340 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Jede Probeflache wurde mit dem Pantometer gemessen, weil
sie hier zu uneben war, um die Winkelspiegel zu benutzen. Die
Flachenform nahm ich moglichst vicreckig, zuweilen aber fiinf-
oder sechseckig.

Die Auswahl der Probeflachen geschah gemeinschaftlich,
wobei sich die Abiturienten unseres Forstinstituts betheiligten.
Obgleich wir nach einem bestimmten Arbeitsplan arbeiteten, so
revidirten wir doch immer die fertiggestellten Probeflachen, um
Gewissheit dariiber zu haben, ob auch alle Arbeiten correct
ausgeftihrt waren. ;

Die Aufnahmen der einzelnen ausgewahlten Probeflachen
geschah nach dem R. Hartig’schen Verfahren, bei welchem
Probe-Stémme gefallt werden, weil wir dieses fiir das richtigste
hielten. Die Durchmesser der sdmmtlichen Stamme der Ver-
suchsflachen wurden in 1.3 m. Hohe vom Boden in Abstu-
fungen von I cm. zu I cm. kreuzweise genau gemessen. Die
Messhohe, nahmen wir immer an der Baumaxe, das. heisst
bei einem Bergabhang nicht von oben und nicht von unten
gemessen, sondern in der Mitte von beiden.

Je 3 Probestamme wurden von jeder Probeflache ausgewahlt,
welche in der Hohe von 0,3 m. vom Boden gefallt wurden. Der
verbliebene Baumstock wurde in seiner Mittelstarke (also bei
0,15 m. von Boden) dreifach kreuzweise gemessen, um spater
die Stockholzmasse berechnen zu kénnen. Die gefallten Pro-
bestamme wurden erst I m. lang vom Abhieb, dann in je 2 m.
langen Sectionen, deren mittlerer Durchmesser bis auf Millimeter
genau doppelt itiber’s Kreuz abgegriffen wurden, kubirt, das
Astholz jedes Probestammes anfanglich gewogen und xylome-
trisch kubirt und erst nachdem die Verhaltnisszahl zwischen
Gewicht und Volumen festgestellt war, erfolgte die Kubirung
nur noch mittelst Wagung.

Zur Darlegung der Bestandesaufnahme fiihre ich folgendes
Beispiel an:

Probeflache No. 42.

Schulwald zu Kiyosumi; Ortsname: Ippaimidzu; Abth:
I; Unterabth: C; MeereshGhe: 250 m.

Die Probeflache-Aufnahme vom 2 bis 10 December 1894
geschah durch die Herrn Horimoto, Mimura, Okuda, und
Matsudaira ;

8.8984

PROBEFLACHE Ny. 42,

el a 8.3984 ‘
| e= us d=29,0cm, M= Digeren X 9237 Jahrringszahl : 70°
E Yo) 14 qm. ot
6 FOOSiT4 g=0,066052 qm. =117,4463 fm,
- Alter: 7o+2
m=0,9237 fm. = Ss Co s712
d= 28,4 cm. ea fm Letzte 5 jahrige
; a@=0,05712 fm. , : Triebslange : 79 cm.
5 h= 26,3 m.
5
7
uf
2
i
6
16
30 5 | 3534
31 10 | 0,7548
(32 4 | 09,3217
33 8 | 0,6842
34 0,6355
35 5 | %48rr
36 10 1,0179
37 5 | 95376
Sa. | 133 | 8,3984 |
ahs 3 | 0,3226 |
132 | Faye
38 - a es d=41,5 cm. vt =a x 1,7829 Jahrringe : 70
39 Ir 1, 3140 =0,13361 &=0,1353 qm. = 109,1628 fm. Alter: 7042
40 3 90,3779 A= Se er |
ane meniesio2q2z | Sa aaa a x 0,11 70 Letzte 5 jahrige
42 4 | 05542 a=0,1170 fm. =7,1546 fm. Triebslange : 100cm.
43 7 1,0165 6
44 2 | 0,3041 Bae?
45 8 | 1,2723
| ; 46 4 | 0,6648
{i Nae ET | 91735
| |) Si, 62 | 8,284r
ie 4 a :
7 11735 __ 83577 d=53,6 cm. M =3,26009 X 37 Jahrringe: 70
0,7238 3G as
=120,6200 fm.
1,1314 | =0,22588 m=3,2600 fm. Alter: 704+2
0, 4036 Joe A =0,22556 X 37 a oe
0,4247 Biers ce a@=0,22556 fm. =8,3462 fm. Letzte 5 jahrige
Triebslange: 85 cm.

PHP PDK AWARKDND APD

H

h=31,4 m.

M=347.2291 fm.

A= 22,7630 fm.

a, A=Astholzmasse ; m, M=Schaftholz.

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HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 341

Bestandsgrosse: 1 ha.

Probeflachengrésse: 0,2866 ha.

Der Grund besteht aus lockerem Sandstein; Die Boden-
flache ist nach Siidosten um 30° geneigt und ziemlich trocken.
Bestandsschluss: gedrangt.

Der Bestand wurde von jedem Kenner als einer der besten
erklart.

Hieraus ergeben sich pro ha folgende Verhaltnisse :

Alter, das Mittel aus den mittlern Altern der Klassen sowohl,
wie aus den Massenaltern: 72 Jahre.

Baumhohe: oberere 31,5 m. mittlere 28,6.

Baumstarke: starkste 63,0 cm. schwachste 15,0 cm. mittlere
37,0 cm.

Stammzahl: 810

Holzmasse: Schaftholz = 1211,55 fm.

Astholz = - 79,426: ,,
im ganzen = £290,970" ;,

Stammgrundflachensumme: 87,376 qm.

Griines specifisches Waldgewicht des Schaftholzes im
Durchschnitt 0,734.

B. Ergebnisse der Bestandesaufnahmen.

Da die Sugi meist nur auf guten Boden gepflanzt wird, so
war es sehr schwierig gewesen, viele schlechtere Bestande zu
finden. Bei unserer Untersuchung hatten wir neben 32 Probe-
flachen bester Giite nur 24 Probeflachen geingerer Giite, unter
denen allerdings auch solche mittlerer Giite vorhanden waren.
Wir lassen nun das gewonnene Resultat tabellarisch folgen
GBab., 2),

342 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

IV. Konstruktion der Ertragstafeln.

a. LEntwurf der Holzmassen oder Zuwachscurven.

Nach der Aufnahme simmtlicher Versuchsflachen im
Walde und Ausfiihrung der zugeh6rigen Berechnungen schritten
wir zur Konstruktion der Holzmassencurven, wobei Schaftholz
allein oder Schaft- und Astholz zusammen zu betrachten ist.

Zu diesem Behufe wurde auf ein 70 cm. breites und 110 cm.
langes Blatt Millimeterpapier eine horizontal gezogene Linie
(Abszisse) in 110 gleiche Teile geteilt, weil der aufgenommene
alteste Bestand nur 108 Jahre zahlte. Auf den einzelnen
Teilungspunkten dieser lLinie, welche die Bestandesalter
darstellen, wurden Senkrechte (Ordinaten) errichtet, auf diese
die in den einzelnen Versuchsflachen gefundenen wirklichen
1 fm. zu + mm” auf-
getragen und die Enden der Ordinaten mit kleinen Punkten
versehen. Wir erhielten so auf dem Papier, so viel Punkte, als
Versuchsflachen aufgenommen wurden; denn keine zwei Punkte
deckten sich vollkommen. Die einzelnen, die Masse im jugend-
lichen Alter darstellenden Punkte standen natiirlich naher bei

ce

Bestandes-Massen in einem Massstab

einander und entfernten sich mit wachsendem Bestandesalter
von links nach rechts aufsteigend strahlenformig immer mehr,
weil in jiingeren Bestanden die Massen resp. Massendifferenzen
zwischen den besten und schlechtesten Standorten wesentlich
geringer, als in alteren Bestanden von 70 oder 80 und mehr
Jahren der Fall ist.

Um nun bei Ausscheidung von vier Bonitaten die vier
Massencurven zu erhalten, zogen wir zunachst vom Jahre Null
ausgehend, durch die hochsten und ebenso durch die niedrigsten
aufgetragenen Punkte, oder moglichst nahe an denselben voriiber,
aus freier Hand je eine Linie, wobei kleinere Unregelmassig-
keiten, wie sie bei durchschnittlich zu grossen oder kleinen
Massen vorkamen, unberiicksichtigt blieben. Aus diesem
Grunde konnte auch die obere und untere Linie nicht alle
Punkte mit einem Curvenzug durchsetzen. Die obere Linie
stellt dann ungefahr die obere Grenze, die untere Linie die
untere, der in verschiedenen Lebensalterh der Bestande vor-
kommenden Massen dar. Die Genauigkeit dieser Mittelwerthe
wird natiirlich um so grdsser sein, je mehr Versuchsflachen

TAB. I. UBERSICHT

der in den einzelnen Versuchsflichen normaler Sugibestdinde wirklich erhaltenen Ergebnisse. (pro. ha.)

3 ; 3 2 9 5 P
3 s g ma Durchmesser, Schaftholz. pebats Bn SEUODZ, Ps alot = cin | fee >}
& aes Exposition 5 nae : a masses ir E She 55 mgt w'| perme eeu eB
I Be sh il Is P 8 a z ee | ee Bl Ben se\Uee\e2 | 5
5 Oe o “cs H $ a a a0 N&e| et. | a@en|/8eS) ae <
Zs ORTSNAME. | 7% § | und Z| » |3 |e 3 E 5 Ea Re 5 Zn 2¢ |22s| SSE /EESIERR 28]
3 3 | & & | 3 & . (Sapam 3 & | seh5| on Nie esi. a0 BOE| APS |S3a 0/45 %5| 9 w Z
4 ue 4 wv | oO S SE las& | se S Re SSE ce Cine) 9agE el a oU5 | o 8 wswoleos] « §
& oe gv Nei wo] 8B | Se eas Zo S10 || Stee 23a a dos SeESte ae recat) she FEn| bao oie (ae
a a je) oe eo eT eee Baie oe Eyes | 22 ieee se Pe |aeel em
E =I ]
|
1 | Matsuba..........| O,11520] 300| S.O. 302)|| TO! 351 2520) meres 2,4 4,9 | 2674 19,57 1,957 17,216 36,786 3,679 5,081 | 3,74 0,0019 | 0,05 | 87,9 | 0,665 | IV
2 ” ee eeeeee e+| O,25000 | 400 | N.O, 15°| 11! 7,5 | 5:53] 19,0] 3,0] 6,0} 5215 64,00 5,818 | 38,591 | 302,591 9,326 14,601 | 1,92 0,0028 | 0,15 | 60,3 | 0,820 I
3 | Tokansawa_ ......| 0,25000] 348 | S.O. 29°| 12] 3,5 | 2,80] 11,0] 1,0] 2,0 | 2080 11,84 0,987 5,870 17,216 1,435 0,676 | 4,85 0,0003 | 0,01 | 45,4 | 0,680 Vv
4 |Imasumi..... + + +++] O,20000 | 350 Ebene I5| 11,0] 9,61] 21,0] 4,0] 11,2 | 2740 136,80 g,120 595539 | 196,339| 13,089 27,123 | 3,65 0,0099 | 0,27 | 43,0 | 0,863 1
Fallen De terrctreate’eletat sic 0,28600 | 300 | N. 5o|| £7) L254.| 7,20)|20;0 2,0 | 11,3 | 2788 92,15 53420 53,368] 145,514 8,560 27,745 | 3,59 0,0100 | 0,28 | 57,9 | 0,791 | III
Ga) Karidoshin terse «are 0,07420| 150] E.S, 12°| 17] 16,0 ]12,00] 21,0 4,0 | 10,9 | 3666 198,90 | 11,699 57,921 | 256,821 | 15,107 34,304 | 2,72 0,0094 | 0,34 | 15,5 0,852 I
7 ” oe ++eeee| O,25000 | 200] N.O. 10°} 20| 15,2 | 11,52| 19,0 5,0 | 12,7 | 3011 214,00 | 1C,700 | 63,592| 282,592] 14,130 37,820 | 3,31 0,0126 | 0,38 | 3255 0,554 I
3 | beeiiebl oo. coon Apdo 0,19820 | 250| N.O. 5° ||| 22)|| 16,0) | 7,10) x750 4,0 | 10,9 | 2472 85,18 3,872 49,745] 134,925 6,133 23,239 | 3,96 ©,0094 | 0,23] 58,4 | 0,781 | IV
g | Kannonminami....| 0,37622 | 270| O. 12°} 25] 20,5 |15,94| 34,0] 5,0] 17,2 | 2299 | 346,19 | 13,848 | 79,261] 425,454) 17,018 52,483 | 4535 0,0228 | 0,52 | 22,9 | 0,850 I
10 | Imasumi........ ..| 0,17820| 340| N. 8°| 26] 12,3 | 7,52] 19,0] 2,0] 11,0] 5612 | 144,55 5,500 | 36,811] 181,361] 6,975 52,396 | 1,78 0,0093 | 0,52 | 26,4 | 0,863 | IV
TT |e Mlatsu bay areteretaisiels ers 0, 38360 | 321 | O. 34°,| 28] 20,4 | 14,60] 40,0 3,0 | 17,0 | 1741 267,91 9,568 | 41,602] 309,508] 11,054 40,518 | 5,74 0,0233 | 0,41 | 15,5 | 0,880 i
120) Gran Min O) estalelel=iere 0,04278 | 288 | S.O 32°} 30] 13,5 | 11,30] 23,0 | 3,0 | 10,2 | 5426 312,92 | 10,431 | 129,437] 442,357] 14,745 45,104 | 1,84 0,0083 | 0,45 | 41,4 | 0,62
13 1 fe eeecnss 0,08g910 | 288 | S.O. 30°| 30] 16,4 | 14,00] 30,0} 3,0] 17,1 | 2330 | 430,95 | 14,400 | 104,319| 535,269| 17,842 53,657 | 4,29 0,0230 | 0,54 | 24,2 | 0,760 iv
14 m sapoeoc .+»| 0,25000 | 280] S. 20°| 35| 12,1 | 9,45] 28,0] 5,0] 12,0 | 3470 220,00 6,286 48,123 | 268,123 7,661 39,126 | 2,88 0,0113 | 0,39] 21,9 ee Hi
15 | Kannonminami....| 0,10140| 273 | O. 20°| 36] 21,0 | 16,30} 36,0] 3,0) 18,4 | 2071 461,48 | 12,819 | 97,557) 559,037] 15,529 54,826 | 5,24 0,0263 | 0,54 | 21,1 | 0,6
5 I
16 | Imasumi..........| 0,28940]| 340| O. 10°} 36] 23,0 | 20,67] 43,0 | 9,0 | 22,0 | 1921 685,60 | 19,044 93,722] 779,322| 21,648 75,704 | 5,21 0,0394 | 9,76] 13,7 | 9,790
17 | Banshomai........ 0, 16800 a O. 20%25°| 36] 23,0 | 19,52| 50,0] 4,0] 23,0 | 1547 | 538,03 | 14,940 | 95,736] 633,766] 17,605 64,442 | 6,46 0,0423 | 9,64 | 17,7 ses ial
18 |‘Tokansawa ...... 0,11288 | 280| S.O. 25°| 36] 14,1 | 12,60] 35,0] 4,0] 16,5 | 2374 | 304,95 | 10,970 | 69,135) 464,085] 12,891 50,0g0 | 4,21 0,0211 | 0,50) 17,5 or775 ;
19 | Yedosili ...... ....| 0,14884 | 276 | S.O. 9°| 38] 22,4 | 20,30] 40,0) 3,0] 21,0 | 1841 | 610,04 | 16,062 | 60,360) 670,400) 17,642 63,585 | 5543 0,0345 ae 9,9 Bi 8 if
20 | Banshomai........ 0,15 442 | 333 | O. 22°| 38] 20,2 | 18,50) 34,0] 3,0 | 19,7 | 2163 | 557,32 | 14,666 | 49,967| 607,287/ 15,981 65,721 | 4,50 | 0,0304 | 0,66) 17,0 | %7
OH 7 86
21 | Okudari Higashi ..| 0,2 220| S.W. 14°-30°|} 39] 24,7 | 20,14] 38,0 | 10,0] 23,7 | 1660 749,59 | 19,220 98,550| 848,140] 21,747 73,359 | 6,02 0,0440 | 9,73 | 4351 By 4
22 | Kudagara ie Aanciod Sayane 200 | E.N : ae 40 eee 11,02] 27,0] 10,0 | 17,1 | 1870 347,74 8,694 | 116,075) 463,815] 11,595 43,312 | 4,31 pee sae a3 yes a
2awiliohobudaremrecrcderiee 0,14820| 288 | N.O 3°| 43] 26,0 | 20,15] 54,0| 6,0 | 31,5 | 1053 | 697,10 | 16,210 | 107,207] 804,307] 18,705 81,749 | 9,50 0,077 es 2 hs Reed 1
2au| Wishimitsul sejetvieteree 0, 08016 | 260| O.N. 15°| 45| 19,0 | 18,02] 23,0] 3,0 | 21,3 | 2146 | 595,53 | 13,230 | 112,969] 708,409] 15,744 75,952 | 4,66 010355 oP | =e Ces I
250|Okuboana) wees cere 0,14284 | 220| O. 39°| 46] 25,0 | 21,06] 48,0] 8,0] 22,1 | 1988 | 767,30 | 17,333 | 134,383 | 931,683 | 20,254 76,033 | 5,03 0,0382 Wi) |e , *
26 | Shobuda.......... 0,11600 | 288 | O.S. 2 | 948 |) 25,0) 8 yum eon mone zere) | 2483.) ©30,49,|/ 43,320 |, 68,801) 7oBs350 | T4757 | b2050 | ae | arte es ae Alec 1
27 | Mukomine ........| 0,13660| 170| W. 4o°| 48] 22,1 | 21,20] 56,0] 8,0 | 23,1 | 2218] 858,23 | 17,464 | 49,322 997,549 18,907 ooes ahs ae 0,69 | 33,0 | 0,780] IL
28 |Sakurago ........ 0,25576| 261 | O.N. 24°| 51| 24,4 | 22,86] 59,0] 3,0] 33,4 | 1572 675,61 |. 13,250 | 228,042] 903,652 ee pees pe pate a tye ny Pe IV
29 | Minamisawa ...... 0,18240| 150| OO.S.29°-30°| 51| 17,0 | 13,80] 30,0} 2,0} 14,8 | 2719 296,27 5,809 47,704 | 344,034 hee 46,629 ate ae 7 aa! na ae IV
BOM |ilimastinaiere eee 0,13840| 320| N.O. 5°| 53] 14,8 | 12,30| 24,0] 2,0] 17,0] 4798 | 428,60 8,087 | 76,798] 505,396] 9553 110,790 , 3023 ) , :
| 2 | 0,81
31 | Banshomai........| 0,13566 | 330| O. 30°| 54] 25,0 | 23,60] 46,0 5,0 | 31,4 | 1887 972,22 | 18,004 118,533 | 1090,753 | 20,199 ae Bee oe | oa wae agae i
32 | Banshoshita ...... 0,37166 | 200| O. 38°| 56] 28,0 | 25,80| 51,0 | 10,0 | 27,6 | 1512 | 1012,54 | 18,081 | 132,132] 1144,672| 20,441 9925 Pee ee 079| 5,9|0,793| J
33 Ailes), aeeise oe 0,21154| 250| N.OO 35°| 57| 28,8 | 25,30] 57,0 | 13,0| 32,3 | 967] go1,05| 15,808 | 52,738) 953,788 ye. 7 be he aes ok 13,2 0,852 II
34 | Gomondimukai ....| 0,14768 | 320] S.O 25°| 58] 22,6 | 21,10} 42,0] 4,0 | 23,4 | 1885 791,58 | 13,648 | 99,743 sone 15,3 ee 4 aos ines 0 88 | 8,9 | o,79r Il
35 | Minamisawa ......| 0,12428| 180] O. 22°| 59| 24,8 | 23,80] 43,0] 6,0] 24,5 | 1865 | 948,47 | 16,076 | 84,438 | 1032,908| 17,507 3079 | 43 3047 , ( ;
0,800
36 0 ahersie 0,20000 | 180 | N. 20°| 61] 30,2 | 26,92] 50,0} 10,1 | 31,0 | 1121 | 1054,12 | 17,281 | 116,512 Melee pose ee ae eee ne as eS eeoyLIN
37 | Matsuba........../ 0,47012| 300/ W. 30°-40°| 62] 26,5 |17,21| 63,0] 3,0] 29,9] 842] 578,69 | 9,334 | 75»743| 9541432 101555 Paene 12,59 | 00855 0,67 | 9,6 | 0750 MI
38 Sl winsadoweus. 0,10776 | 330| S.O. 31°| 64| 27,0 | 19,00] 59,0] 15,0 | 33,0] 780] 721,34 | 11,271 | 68,960 ik ee 6. - 3,20 eee 0,37 | 20,0 | 0,730| V
39 | Riumensawa...... 0,10400 | 280| N.W. 21°| 64) 14,6 | 12,05] 26,0) 2,0) 12,0 | 3173 217557 3,399 445290 ae 2 Veee 4 a2 4,90 0,0312 0,63 17,1 | 0,709 IV
4° ” e+e+..| 0,24266 | 280| N.W. 25°| 68) 17,5 | 15,70] 48,0] 4,0] 19,9 | 2028 491,67 7230 84,454 | 576,124 247 3,253 , , . ae te
Oo, 14, 0,59
41 |Sakurago ........| 0,13340]| 220| S.O. 42°| 68] 19,1 |17,05| 46,0] 6,0] 22,0] 2039] 554,00] 8,150 | 79,870 Sastre Biba gubse ee, ares ote ae G34 1
42 | Ippaimidzu...... ..| 0,28660 | 250 | S.O. 30°| 72]| 31,5 | 28,60] 63,0] 15,0] 37,0] 810] 1211,55 | 17,064 | 79,426 Theres oBee UG I eo 0,0826 | %77| 45,3 | 0,870) Il
43 | Tobikoshi ........ 0,26000 | 300 | $.0. 18° 73 || 2655 |/21;20| Rayos eawioniegg;0)| 520.) 751,03 | 1,288. | 1141575) "Por 5 | Ehree eo sake 0,0879 | %85| 6,1 | %713| It
44 | Koyagao...... +++] 0,29320| 180/ S. 7°| 73| 32,1 | 28,04] 60,0] 18,0} 33,5 | 968 | 1105,70| 15,150 | 67,720) 1173,420) 1 ates ae 11,28 | 0,0897| %79| 90] %740| Il
45 |Ushirosawa ......| 0,15794| 228| N-W 20°| 75] 29,5 | 24,30] 70,0] 9,0] 33,8| 899| 854,79; 11,397 | 76159] 939,949| 12,413 793499 , 5 i Ee leet
46 | Koyagao.......... 0,31528 | 180 | O. 20°| 75| 34,1 | 26,30] 64,0] 6,0] 37,r] 755 | 925,15 | 12,335 | 59811| 984,061) 131133 Oe daeae OES o75| 6,6 | 0,810 | 1V
47 | Jizodobori ........| 0,13374 | 250| E.N. 35°| 75| 20,0 | 20,60] 42,0} 8,0| 24,4 | 1609 | 659,23] 8,789 | 42,459) 701,689| 9,356 | 74,807 | 2! o1056 | 088 | 8,3 | O72 I
48 | Ushirosawa ...... 0,24610 | 230| W. 34°| 77| 327 | 29,61] 50,0] 17,0] 36,7] 832 | 1194,80] 15,516 | 98,512 | 1293,312| 1 a acts 16 0,0457 0,84 9,3 | 0,704 | Ll
49 | Tokansawa ......| 0,15620| 220| S. 35°| 79| 26,0 | 20,68} 50,0] 9,0] 24,0 | 1831 779559 9,867 | 72,503| 852,003 ae es ay 0,0435 0,90 | 13,3 | 0,880] IV
50 | Koyagao.... ssc. 0, 13340 180| SS.0.35°-40°} 81 | 25,2 | 15,00] 54,0] 3,2 | 23,6 | 2068 708,21 8,743 92,314 | 800,524 9,553 9925 4) ’ Bete +
7 0, ) i
51 | Tokansawa_ ......| 0,37600| 220| N.O 42°| 84| 28,0 | 25,20] 60,0| 5,0| 31,6 | 1213 | 1068,54 | 12,721 | 61,421 | 11209,961| 13,452 95,366 soe ied fe ae 0,670 I
52 3 + +++. 0,40000 | 220] N.O. 25°| 84| 33,5 |30,30| 61,0] 28,0] 39,0] 761 | 1246,10 | 14,834 | 93,250] 1339,350 151945 9 a 3) s 0,0516 0,94 | 19,7 | 580 | 1V
53 |Ippaimidzu ......| 0,39600| 320| S.O 32°| 95| 28,0 | 19,90] 67,0] 3,0] 25,9 | 1818 | 797,55 | 8,395 5p Bee Pee ee Ee? site 0.1324 | 0,93| 65 | 0630] 1
54 ” ++ +++] 0,50000 | 320] N.O, 15°] 95| 35,2 |32,11| 72,0] 25,0] 41,1 | 7or | 1303,00] 13,715 | 84,12 3 ora a - Ay a ae 0,0536 | 068 | 12,4 | 750 IV
55 ” ++ ++++| 0,30000 | 320 | S.O. 20°] 99| 24,2 | 21,10) 80,2 | 21,0] 26,1 | 1275 | 745,81 7533 92,451 35,2 i 4 7 ae 9 plies 011788 071 | 757 0,800 | Lik
50 Tobikoshi ........]0,30106 | 300 | N. W. 23° | 108| 31,5 | 25,50] 74,0| 26,0| 48,0] 399 | 1002,82 | 9,285 | 76,300 | 1079,12 9599 715357 | 25)

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HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI 343

bester und schlechtester Standortsgiite aufgenomen wurden und
je gleichmassiger sie sich tiber die einzelnen Altersklassen
verteilen. Da die Massen einiger aufgenommener Versuchs-
flachen auffallend unter der unteren Grenzlinie liegen, so scheint
es mir, als wenn an dem betreffenden Orte wohl noch niedrigere
Werthe vorgekommen waren ; es war aber nicht moglich gewesen,
eine noch geringere Massenklasse, also 5 te Bonitatcurven zu
konstatiren, weil in diesen Gegenden Swgi nur sehr selten in
schlechtem Boden gepflanzt wird. Solche Ergebnisse blieben
daher unberticksichtigt.

Wir wussten nun, in welchen Grenzen sich die Ertrags-
verhaltnisse der Bestande tiberhaupt bewegen. Da es wiinschens-
werth ist, die einzelnen Bonitaéten gleich weit von einander
abstehen zu lassen, so theilen wir jetzt bei der graphischen
Darstellung die Flache zwischen der unteren und oberen
Grenzlinie der Lange nach in vier gleiche Streifen. Die
Theilungslinien laufen natiirlich alle im Nullpunkt zusammen
und die Streifen werden um so breiter, je mehr sie sich von dem
Nul!punkt entfernen wie es auf der P]. XVIII ersichtlich ist.

Alle Punkte, resp. Besténde, welche nun in den obersten
Streifen fallen, k6nnen als zur I., solche im zweiten Streifen als
zur II. etc., diejenige des untersten Streifens als zur IV. Bonitat
zugehorig betrachtet werden.

Wir haben zuerst bei der Bestandesaufnahme nur beste und
schlechteste Bestande ausgewahlt ; jedoch war die Auswahl hier
nicht ganz nach Wunsch, weil in der Regel die Bestande mittlerer
Bonitat reichlicher vertreten waren und weil auch die vorher
nur nach dem Augenmaas in ihrer Bonitaét eingeschatzten
Versuchsflachen sich theilweise bei der Construction der Mas-
sencurven noch etwas andersergaben. Viele in die I Bonitat ein-
geschatzte Bestande fielen nach dem Auftragen in den Streifen
der II, auch viele in die IV Bonitat geschatzte Bestande fielen
nach dem Auftragen in den Streifen der III.

Bei den Streifen der IV zeigen sich noch einzelne Liicken,
welche wir wohl spater bei weiteren Untersuchungen beriick-
sichtigen werden.

Nachdem so die Normalertragskurven konstruiert sind, so
ist es leicht mittelst der Millimetermassstabes auf Pl. XVIII
und XIX die jeder Bonitat und jedem Bestandsalter entspre-

344 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

chende Holzmasse abzulesen und in die Ertragstafeln einzutra-
gen.

b. Entwurf der Hohenzuwachscurven.

a. Die Kurve der Bestandsmittelhohe.

Nach der Ermittelung der Ertragsverhaltnisse haben wir
die zu jeder Bonitat gehorigen Versuchsflachen unterschieden.
Mit der zur I Bonitat gehodrigen Mittel-Hohe construirten wir
die Bestandmittelhéhencurve in der Weise wie bei den Holz-
massenzuwachscurven; in gleicher Weise auch die IV Bestandes
mittelhdhecurven von der zur IV Bonitat gehdrigen Versuchs-
flache.

Wenn in sammtlichen Versuchsflachen bei den Probestaém-
men auch die Hohen der Langentriebe der vorhergehenden 5
Jahre gemessen werden, so erhalt man auf diese Weise auch die
mittleren Bestandeshohen vor fiinf Jahren, so wie durch An-
rechnen der letzten fiinf Langentriebe zur gegenwartigen Hohe
auch die muthmassliche Hohe nach 5 Jahren. Auf diese Weise
schliessen die einzelnen aufgetragenen Ordinaten viel enger an
einander, was namentlich erwiinscht ist wenn gréssere Liicken
in den Beobachtungen vorhanden sind.

Aus der graphischen Darstellung von der I u. IV Hohen-
curve ergiebt sich auf graphische Weise die II und III Hoéhen-
curve wie auf Pl. XX ersichtlich ist. Aus diesen Curven folgt
mittelst Millimetermaasstabs jede Bestandsmittelhohe zu jedem
Jahre und diese wurde in die Ertragstafel eingetragen.

8. Die Curve der Bestandsoberhohe.

Auf ganz analoge Weise fanden wir auch leicht die Bestands-
oberhohe aus wirklich gemessener Bestandsoberhohe.

c. Entwurf der Kreisflachencurven.

Obgleich die Bestandeshdhe als der wichtigste Maasstab ftir
die Beurteilung der Standortsgiite bekannt ist, so ist doch auch
die Kreisflachensumme des Bestandes auf der Flacheneinheit
(Hektar), bezogen auf 1,3 Meter tiber dem Boden, und ermittelt
fiir alle Bestandesalter und Bonitadten, namentlich dann von
Werth, wenn es sich darum handelt, die Holzmasse eines

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 345

konkreten und nicht tiberall normal bestockten Bestandes mit
Hiilfe von Ertragstafeln rasch und ohne Fallung von Probe-
stimmen zu bestimmen. Hatte z. B. der auf seine Masse zu
untersuchende konkrete Bestand nur 0,75 der Kreisflachen-
summe des Normalbestandes der Ertragstafel, Hohe und Alter
waren aber in beiden gleich, so wird auch ersterer nur 0,75 so
viel Masse pro ha als letzterer haben, d. h. die Holzmassen sind
in diesem Falle proportional den Kreisflachensummen. Ausser-
dem ist auch fiir Wirthschaft und Wissenschaft die Unter-
suchung der Kreisflachenmehrung normaler Bestande von deren
Begriindung an bis zur Haubarkeit nicht uninteressant. Ich
brachte daher in Pl. XXI auch die Kreisflachensumme fiir die
einzelnen Bestandesalter und Bonitaéten graphisch zur An-
schauung.

Es wurde hierbei genau so wie bei Bestandshohencurven
durch Auftragen der wirklich gefundenen Kreisflachen verfahren,
welche nach den Bonitaten eingeteilt waren.

d. Entwurf der Stainmzahlcurven.

Bei Begriindung eines Bestandes ist nattirlich die Stammzahl
pro Flacheneinheit in den ersten Jahren am _ groéssten, aber
allmalig breiten sich die einzelnen Baume aus, die Aeste kommen
naher zusammen, es entsteht ein Kampf ums Dasein, der zum
Tod der schwdcheren Exemplare fiihrt. Die Stammzahl pro
Flacheneinheit nimmt deshalb von Jahr zu Jahr ab. Wahrend
wir in einem angepflanzten Sugi-Bestand am Anfange der
Umtriebszeit pro Hektar 6000 Pflanzen gezahlt haben, sind
am Ende derselben kaum noch 500-600 Stémme vorhanden. Es
ist interessant und von praktischer Wichtigkeit, das Gesetz der
Stammzahlabnahme fiir alle Jahre der Umtriebszeit festzustellen.

Nun wurden die Stammzahlencurven fiir jede einzelne
Bonitat aufgetragen nach Altern als Abcissen und die Stamm-
zahlensumme als Ordinaten. Die Werthe derselben lasen wir
fiir jedes fiinfte Jahr ab und trugen sie in die Ertragstafel ein
(Siehe Pl. XXII).

e. Sonstige Bestandteile der Ertvagstafeln.

Unsere Ertragstafeln enthalten weiter noch den laufenden
und durchschnittlichen Hohenzuwachs, den laufenden und

346 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

durchschnittlichen Zuwachs des Schaftholzes, sowie des Schaft-
und Astholzes, Durchmesser des Bestandsmittelstammes, Be-
standsformzahlen, Bestandsrichthohen, Zuwachsprocent vorwarts
und endlich den Normalvorrath und das Nutzungsprocent.

Der laufende Zuwachs ergiebt sich aus den Differenzen der
zwei aufeinander folgenden Gliedern der Tafeln.

also z. B. 80 Jahr, Masse 1229 fm.
85 ” ” 1257 o>
mithin Zuwachs fiir 5 Jahre 28 fm.

Wenn dann angenommen wird, dass innerhalb des Jahrfinftes
der Zuwachs jairlich gleichmassig erfolgt, so wiirde derselbe in
dem vorstehenden Beispiele pro Jahr = 5,0 fm. betragen.

Der Durchmesser des Bestandsmittelstammes wurde
gefunden durch Division der Kreisfléchensumme durch die
Stammzahl.

Die Bestands-Formzahlen ergeben sich aus den bereits
bekannten Werthen:

g Kreisflachensumme,
h Bestandsmittelhohe,
m, Schaftholzmasse,
m, Gesammtmasse.
Die Schaftholzformzahl ist aa
Die Gesammtbestandsformzahl

Mz
7]

Die Bestandsrichthéhen ann man entweder erhalten,
indem man die Hohe und die Formzahl] mit einander multiplicirt
oder die Masse durch die Kreisflache derselben dividirt. Beide
Wege fiihren zum Ziel. Die Berechnung aus h und f diente
dann als Probe fiir die Richtigkeit, denn, sind die auf beiden
m

Wegen erhaltenen Werthe richtig, so muss h. f.=—— sein.

oD?

Bei dem Zuwachsprocent ist als Capitalstock die Masse im
je fiinften, also 5, 10, 15 u.s.w. Jahre angesehen, als Zuwachs
aber die jahrliche Vermehrung des nachsten Jahrfiinftes.

Haben wir also z. B. das Zuwachsprocent fiir den 30 jahrigen
Ort I Bonitat zu berechnen, so ist die Masse im 30 Jahr=443
fm.; vom 30 zum 35 Jahre erfolgt jahrlich 24,6 fm. Masse,
mithin haben wir ein Zuwachsprocent

24,6x100 __ 2460 __
443 443. 9999S"

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 347

Der Normalvorrath ist nach der Pressler’schen Formel
n(a+b+c+ Biers oe berechnet, wobei » die Zahl der
Jahre, hier also 5 bedeutet, in a, 6, c, aber die Massenangaben
der Tafel fiir das 5, 10, 15 etc. Jahr bedeuten.

Die Formel giebt den Vorrath des Normalwaldes fiir das
Frihjahr an. Der Fudeich’schen Auffassung folgend ist dieser
Vorrath als der eigentlich normale angesehen; die Vermehrung,
welche innerhalb eines Jahres erfolgt, ist Jlediglich als die
Verzinsung des Kapitalstockes zu betrachten. Durch die
Nutzung wird sie absorbirt und das Capital stets auf seine
Anfangshohe zurtickgefihrt.

Fiir das 30 te Jahr der Bonitat I berechnet sich sonach der
Normalvorrath an Gesammtmasse, wie folgt :

@ = 20 im. Vorrath int 5 jalhte
i= WA) op if aELOY es
C= ers) Ge 3 aS Car ae
222i, * 59 LOS yp
€ = 327 an ne. 9» 25 3)
SIF = ZA) 9 9» 30 ”

Sa 982,5 fm. Vorrath. Da n=5 Jahre, so ist der
normale Vorrath 5 x 982,5—221,5=4691 {m.

Diese 4691 fm. lassen jahrlich eine Nutzung von 443 zu,
mithin ist das Nutzungsprocent = “= =9,44 %

348 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

TAB. II. NORMAL-ERTRAGSTAFELN

|
; | Bestands-
4 o) Bestands, Holzmasse. :
: Se lives) 2 formzahl.
les} Oo w E =|
E | ee |eee
> eo om
~ < Ts Bc | fm.
a N 5 € OF Re 2 o | N Ke
HH = & air = a =>
za = ee, | oka oe 3 2 Eo
= ree OF 62 v a el z 0 en ee
< mn ie Eas o .& 6s & Re Ss Pe Sg
> ist & OF 35] 2 = re} c S a
Ss So ES = 5 Ss 2 52 =I o&
ca an a 2D p= ~ (S) < = & mM oO
QA n N 7

HONDA : ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

FUR DIE SUGI, BONITAT I.

Bestands-

Laufender

Durchsch-

Zuwachs-

ee ee Mt water e|| wre. |
2 2 2 aE olec
Pee PNeiae jes ie jw be F EE
Peewee) S| 6. | Ss. eel eel Be
R 2 Rh e DR 2 R z 5 BS
G co o Oo Z a
— 1} —| 4,0} 4,0| 4,0! 4,0] 42,0] 82,0 40 | 50,0
MAae2,| Saf tO Alte, 2) 10,2) 22:2) 16,9' || * 224 /|27,4
AeOit (0.9 | 13;8:).10,0) $871 02,3 13371 2L,r| 672)19,5
el 75 E0,0)| 20,0 11,1 14,4| 9,6) 82) 1507) 14,7
Mo 6o,9 121212301 753.1 16,2) 7,1) 6,3) 2824) 11,6
7,0| 9,1) 23,2|25,0|14,8/17,8| 5,6] 5,0| 4691r| 9,4
O75 | 10,0) 24.0 20,8) 16,2 19,1 | 4,2) 3,8) 7152) 7,9
B73)| 50,6 | 23,0)/'25,6 17,1 |'19,9| 3,2] 2,9|ro02t8| 6,7
TOE | 11,0) 21,8) 23,4 17,0) 20,3) 2,5) 2,3\23856| 5,7
MEME X2,2|'20,0 (20,8 | 17,9 | 20,3) 1,9) 2,7| 18021 | 5,0
ite )\E 2,0) 17 ,0|10,5 |17,8|20,0| 1,4) 1,2)22656) 4,3
iO | 3,214.0] £3,0|17,5|19,4| 1,1) 0,9|27686) 3,8
Hes) 239,01 21,0) 10,6 /17,0|18,8| 0,9) 0,7) 33042). 3,3
#237 13,6) 9,6) 8,4|16,5| 18,0] 0,7) 0,6|38668| 3,0
£330/14,1| 8,0) 7,0|15,9|17,3| 0,6| 0,4144518| 2,7
13,3|14,4| 7,0) 5,8|15,4|16,6) 0,5) 0,4)50558| 2,4
13,6/14,6| 5,6) 4,8/14,8|15,9| 0,4| 0,3|56759| 2,2
Bee) 04,7| 5,0] 4,0|/14,2|15,2| 0,3| 0,3/63004| 2,0
14,0|14,9| 4,4) 3,6|13,7|14,6) 0,3) 0,2|69548) 1,9
T4,2/15,0| 4,0) 3,0|13,2|14,0] — | — |76108] 1,7

349

Gesammt-
masse.
alee =
Sale| e
Bn ene [Uns
£ Z
40') 5°; | + 5
304 | 33,0] 10
980} 18,9} 15
204 | £30 ZO
3787] 10,7) 25
6073| 8,8] 30
ON 4 | 30
12607| 6,3} 40
16821) 5,4] 45
BI OR en | Oo
26847) 4,1] 55
2482| 3,6] 60
38418] 3,2] 65
44597| 2,8] 70
599972) 255|| 75
57510| 2,3| 80
64183) 2,1} 85
70908] 1,9| go
77849| 1,8) 95
84814] 1,7| 100

350 HONDA : ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

TAB. III. NORMAL-ERTRAGSTAFELN

: Bestands-
oes o Bestands. Holzmasse. -
«afl: de oo ae formzahl.
a) Boece
* ~_ ome VG eo |
= < ss pole |
= NX Pana Bo 2 : | {m. =
= = Bie) fl Ovesurs 3 | sae as +
pl ee te, || eels. | ee = Es
21 2 1 aa S Pees | eee Slee 3 = | §&
< 5 sceeealiecr oN Se Bo 3 = S
mo ao 0 § = a S z a 5 4 o &
7 So Os = = ro. 3
ue ne 3H = o 3) < 3 & mn oO
Q <A N é
Dil ae a = 127, I.70 163 || = S|) == —
ro} — | .— | ==) 3,51 | 5,r04 — 46]; 90) 0 7b aan

£5) 4075 21-70) Geral eo. 20

20| 3646] 32,08] 10,6] 9,14

30| 2826] 50,18] 15,1 | 13,96
35| 2516] 57,80] 17,2 | 15,87
40| 2244| 64,24| 19,1 | 17,60

45| 2005] 69,30] 21,0 | 19,14

50| 1798| 73,99] 22,9 | 20,52
55, 1616)! 77528) cay ez 7

25| 3197| 41,59| 12,9 | 11,76 | 14,87 | 253| 66| 319) O5n7s eeam
60] 1457| 79,62) 26,4 | 22,91
{

65| 1321] 81,27] 28,0 | 23,93 | 26,82 | 931] 104| 1035] 0,479] 0,532
70| 1208] 82,48] 29,5 | 24,85 | 27,73 | 977] 101|-.1078| 0,477 | 0,526
75| 1115] 83,34] 30,9 | 25,603 | 28,57 | 1015] 98} 1113] 0,475} 0,521
80| 1040| 83,95] 32,1 | 26,37 | 29,36 | 1047] 95] 1142] 0,473] 0,516

85| 982] 84,39] 33,1 | 27,01 | 30,08 | 1074] 92 1166} 0,471] 0,513
g0| 938] 84,69] 34,0 | 27,57 | 30,76 | 1097| 89] 1186] 0,470] 0,508

95| 906 84,89| 34,6 28,09 | 31,38 | 1117| 86| 1203] 0,468] 0,504

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 351

FUR DIE SUGI, BONITAT II.

2 Laufender Durchsch- Zuwachs- 5
Beads Zuwachs nittszuwachs procent Schaftholz. Gesammt
richthohe. pro Jahr. pro Jahr. vorwarts. MASSE.
2 2 D 2 He ae elev all, #64
S a N a aS a S a a Be| 3 re} 2
o & ©) & S & © is i an q an sed
& bs: s s s = s s ° oo ° ta | 4
meee ea PES ah) GE ee) ee ee | S| 2a | 3.) =
= 2 a & a & aes & BS Ne Ss NS
° a B) a By} a i) a & 2s & a
n xh Nn ‘B YD S Nn 8 3 Z (5) (o} Z, (3)
oO oO io) oO A Z.
— | —] 3,9] 3,0} 3,0] 3,0] 41,3)81,.3} 30/50,0] 30/50,0] 5
emis] CO, Zit 2. A,01|, 70 on 6.6 167 | 27,5 227 | 23°5)|| TO

4,4| 6,4|10,0/12,6| 6,4) 9,3/15,0|12,r] 497]19,3| 733]19,0] 15
5,2| 6,9)14,4|16,8| 8,4|11,2/10,1| 8,6| 1121|15,0] \1596\14,0| 20

Sree 77 |i 79 2O,2) 10,1 12,8) 755) 6,0) 273n|1r.9| 2903) 11,0] 25
6,9) 8,4|19,0|21,0|11,6|14,r| 5,9| 5,3] 3586| 9,7] 4708] 9,0| 30

78} 9,3 | 20,0 | 22,4/12,9/15,3] 4,5] 4.2] 5532] 8,2) 7052| 7,6] 35
8,6| 10,1 | 20,2 | 22,0| 13,8] 16,2] 3,5| 3,2] 7989| 6,9} 9952| 6,5| 40

9,3 | 10,8] 19,2] 20,6|14,4|16,6| 2,7) 2,5|10941] 5,9|13388| 5,6| 45
NO) 11,4 | 17,6) 15,6|14,7|16,8) 2,1) 139|14357| 5,2117319| 4,9) 50
£075) £0,0)/15,0).15,8|14,8|16,7| 1,5] 1,4|18193| 4,5|22687| 4,2) 55
EL O)||12,3,|12,6)12,4|14,6|16,4| 4,2) 1,1122389| 3,9|26416| 3,71 60

£155 12,7,| 10,8) 10,4 14,3|15,9| 1,0| 0,8) 26882) 3,5|31435| 3,31 65
11,9/13,1| 9,2] 8,6|14,0/15,4| 0,8] 0,6|31629| 3,1] 36696| 2,9] 70

12,2|}13,4| 7,6] 7,0/13,5|14,8| 0,6] 0,5|36590| 2,8| 42156] 2,6] 75
12,5|13,6| 6.4] 5,8)13,1/14,3| 9,5] 0.4|41729| 2,5|47779| 2,4] 80

12,7|13,9| 5.4| 4,8|12,6|13,7| 0,4] 0,3|47018| 2,3]53537| 2,2| 85
13,0/14,0| 4,6] 4,0)12,2|}13,2| 0,4] 0,3|52434| 2,1|59407| 2,0| go
13,1/14,2| 4,0] 3,4]11,8)12,7| 0,3] 0,2/57959| 1,9/65371| 1,8] 95
13,3|14,3| 3,6) 2,8|11,4/12,2} — | — |63580] 1,8|71414| 1,7| 100

352 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

TAB. IV. NORMAL-ERTRAGSTAFELN

Bestands-
formzahl.

Bestands. Holzmassse.

ALTER.
Stammerundflache
bei 1.3 m. in qm.
standsmittelstam-

mes in cm.

STAMMZAHL.

Oberhohe
Schaftholz
Gesammt-
masse

Durchmesser des Be-
Mittelhohe

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 353

FUR DIE SUGI, BONITAT III.
Scone | 2 [ES | Eee | somos | Same

: 3 ¢ ee ee I Se a
45 @ N @ N 3 sy a S RE | 3 mre|
Pee eee eerie bea) ete |e Lee |) &
SA/EG (HEE (SE | EE SEES | $6 |S) $6 | a5| <
Seer ee eo lee ie NE Ce Se oe |. Se
eee Wee jie (Oe le ee ls ee

C) co) o) Caen Se lew, ihe
cae ace l ECiNeLsO|, 1.0| 158) 44 Aone 18 | 50,0 18|50,0| 5
ea ieee 4 Olle) |)e20)|) 5,022 0,070), | Os, 28.2)|) “145 4 5)) TO
Bron 0 5,7)| 1020 8:8)| 4,11 6,3 6,8 i352) 924:| 19,8) .483)| 10,5 |' 15
Mee Orie KOA 24s 5,7 || 7:8 | EA @s0|| . 728ilus5.7 |) L077)| 14,5). 20
Ep eO,o) | 3, Onns O72) .o-2, | C,Ab eg 3) Mite8 | t25)| 2007), 51,5). 25
Ba 75)|125,0)L0,8:| «8,5 | 10,5] 6,5: 5,8.) 2473\|10,3 3330) 9,5]. 30
6,9) 8,3) 16,6| 18,2) 9,6|11,6| 5,0] 4,5] 3909] 8,6| 5087| 8,0] 35
7,8) 9,0) 16,8 18,2) 10,5 12;4| 3,9| 3,6) 5762] 7,3| 7299|-6,8| 40
PONeds7 | L054 27,8 | EL,2)|13,0.| 352) 2,8! Sogx| 6,3) 9962| 5,9) 45
8,9 | 10,3} 15,4. 16,4|11,6/13,4| 2,4| 2,2|10700| 5,4|13056| 5,1| 50
975) 10,8 | 13,8 114,41 11,8 |23,5| 1,8) 1,6) 123738) 4,7|16540| 4,5) 55
HOWE A NTL,O 12.0) 11,8 113/3| 14) 1,3) 57009) 4,1 |20360| 3,9| 60
TOA 17 10,0] 10,4 WE ONS 1 |) Vie r,0) 2073 4.08,7 | 24404.) 355'| 65
Hes 12,1) .8,6)|. 856 |\11,4 02,8 | 0,9|| 0,8 24605) 3,3| 28810). 3,1| 70
MEA NL2,A\ 7 Ae 7-2 TTL 12,4 | 0,7, 0,0:'28677)| 2:91 39357| 2,8) 75
I1,5|12,7| 6,0] 5,8|10,8/12,0| 0,6| 0,5|/32917| 2,6|38070| 2,5] 80
BEy7 52,9) 5,0) 4,8| 10,5) 11,6) 0,5) 0,41 37297| 2,4|42918| 2,3] 85
CHO )}13,%) 4,2 3,8| 10,1 | 21,1) 0,4) O53 | 45794 ZNAGSIO| 25%) GO
EZ, E) 13,5 |) 3,0) 3,2 |) 9,8) 10,7| 0,3) 0,3 46390) 2,0|52923| 1,9|. 95
£253|13,5| 3.2) 2,8) 9,5|10,3| — | — |51072| 1,8|58046| 1,8|100

354 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

TAB. V. NORMAL-ERTRAGSTAFELN

o) o Bestands. Holzmasse. paren
, Ge | Se ormzahl.
Lo )
aE 7c OO ® Wt 0
Cc bed &
% < we Uy
x SS Ses Bie ss 5 fm. ’ ’
rE = ZE oe | & £ S :
4 SI a0 6E a ° 5 ‘s 3
“ ef jeee| g¢ | ¢ ih 2 epee
= an |Ege| Se] BE 2 A = Ee
isa) an a = Pr] a) a vy a c = % &
op) Pa 3 = $ = fo) s Be 2 2 2 o
a —
QA Dn g

on
|
|
|
Ke)
fen)
oe)
Ae)
(oe)
Oo
aN
|
aS
|
|

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 655

FUR DIE SUGI, BONITAT IV.

Bestands- Laufender Durchsch- Zuwachs- Gesammt-
schthoh Zuwachs nittszuwachs procent Schaftholz.
richthohe. pro Jahr. pro Jahr. vorwarts. NUERBIE:

z zZ Beales Milita st: (See si Mobo tl 2
is a is a AS) aN a S Fel 3s ARE nen
2 Soon eSeMnINE cae E e E = a ile les 2 Sah) on
Selec|2e|ee|/Selee|2e) ee] 22 |P2] 22 | F4] =
S = Swe SG =] RSI is} re] =n r=) t=} ee
GB iS =e & <5 & acs iS Se Sse
Coe emits |S 18 ise) & les) B les
a 3 o oY o 3 5 ZO 6 79

o 1o) Oo 1) Z Zz

356 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

V. Gebrauch der Ertragstafeln.

Die vorstehenden Ertragstafeln enthalten unter Voraus-
setzung normaler Bestockung fiir den Hauptbestand pro Hectar
und fiir die Bestandesalter 1-100;

a. Die Stammzahl,

b. die Stammgrundflache bei 1.3 m. vom Boden in Quadrat-
MELEE,

c. die mittlere- und obere Bestandshohe in Metern,

d. die Holzmasse des Schaftes und des Astes,

e. Schaft- und Astholzmasse,

jf. Bestandsformzahl fiir die Schaftholzmasse und fiir die
Gesammtholzmasse,

g. Bestandsrichthohe fiir die Schaftholz und fiir die Ge-
sammtholzmasse,

h. den laufenden Zuwachs des Schaftholzes und des Ge-
sammtholzes,

z. den durchschnitlichen Zuwachs des Schaftholzes und des
Gesammtholzes,

j- das Zuwachsprocent des Schaftholzes und des Gesammt
holzes,

k. den Normalvorrath des Schaftholzes und des Gesammt-
holzes,

1. das Nutzungsprocent des Schaftholzes und des Gesammt-
holzes ;

I. Sie zeigen, in welchem Verhaltniss mit zunehmendem
Bestandesalter die Stammzahl sich vermindert, Grundflachen-
summe und Hohe desselben aber wachsen, und in welcher Lebens-
periode der grésste laufende jahrliche und grdésste durchschnitt-
liche Flachen- und Hohenzuwachs eintritt.

2. Sie geben Aufschluss tiber die mit dem Bestandesalter
zunehmende Massenmehrung an Schaftholz sowohl, als an
Astholz, ferner tiber den Eintritt des Zeitpunktes, in welchem
der grésste laufendjahrliche und grosste durchschnittlichjahrliche
Massen-Zuwachs erfolgt.

3. Sie zeigen ferner, in welchem Verhaltniss mit zunehmen-
dem Bestandesalter die Zuwachsprocente, die Nutzungsprocente
und der Normalvorrath der genannten Sortimente ab- oder zu-
nehmen.

HONDA : ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 357

4. Sie dienen zur Bonitirung konkreter Bestande und ist
vor Allem die mittlere Bestandeshéhe hierbei entscheidend und
zwar wegen der grosseren Bequemlichkeit. Will man z. B.
wissen, in welche Bonitat ein unter mittleren Schlussverhaltnissen
erwachsener Bestand zu setzen ist, so ermittelt man nur dessen
Alter und mittelst eines Hohenmessers dessen mittlere Scheitel-
hohe. Da nach den Ertragstafeln z. B. ein 60 jahriger Sugi-
bestand I Bonitaét 26,49 Meter hoch ist, so wiirde wenn der
konkrete Bestand ebenfalls 60 jahrig und dieselbe Hohe hatte,
er jedenfalls der I Bonitaét angehoren u.s. w. Ebenso wird
irgend ein Bestand zwischen zwei Bonitaten fallen, wenn seine
Hohe, nattirlich immer gleiches Alter vorausgesetzt, zwischen
beide fallt.

Es wird noch ein weit richtigeres Resultat erhalten, wenn
man ausser der Hohe noch die Grundflache ermittelt, weil beim
Gebirgswalde bei gleichalterigem Bestande mit gleichen Héhen
nicht immer gleiche Holzmasse produciert wird.

Die vorstehenden Ertragstafeln werden speciell bei der
Hinschatzung der einzelnen Bonitaten fiir Zwecke der Feststel-
lung der Grundsteuer nothig sein, wenn man bei Bodenunter-
suchungen die Hohe und die Grundflache als Haupt-Faktor der
Standortgiite betrachtet.

5. Sie ntitzen aber auch bei EHinschatzung der Holzmassen
konkreter Bestande, wenn man keine umstandlicheren Bestandes-
schétzungsmethoden in Anwendung bringen kann oder will.
Besitzt z. B. der abzuschatzende Bestand eine volle normale
Bestockung, so wird er auch dieselbe Holzmasse wie der gleich
alte und gleich hohe Bestand in der Ertragstafel haben. Ist
diese Bestockung jedoch keine vollkommene, sondern betragt sie
z. B. nur 0,8 der normalen, so muss nattirlich auch die aus der
Tafel herauszulesende Holzmasse durch Multiplikation mit dem
Faktor 0,8 reduciert werden. Es sind nun verschiedene Falle
moglich :

a. Der Bestand entspricht genau im Alter in der Stamm-
grundflache und Hohe einem Satze der Tafel. Dann gilt die
dort angegebene Masse ohne weiteres.

b. Der Bestand fallt im Alter zwischen zwei Glieder der
Tafel, Kreisflache und Hohe sind aber so, dass sie den Tafelcur-
ven (Pl. XVIII) entsprechen. Dann ist die Masse des nachsten

358 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

jiingeren Bestandes zu nehmen und so vielmal den jahrlichen
Zuwachs hinzuzuzahlen, wie der Unterschied der Jahre betragt
z. B. ein 68 jaéhriger Ort mit 82,00 qm Stammegrundflache und
24,48 m Hohe gehort genau in die Bonitat IT, denn diese hat fiir

65 Jahr 23,93 m Hohe und 81,27 qm Grundflache
7O 9 24,85 oe) 3) ” 82,48 9 ”

Die Masse im.65 Jahre ist 1035 fm. mithin im 68 Jahre, da der
laufende Zuwachs=8,6 fm ist, 1035+3 x 8,6=1060,8 fm.

c. Der Bestand entspricht in Alter und Grundflache der
Tafel, aber nicht in der Hohe; dann wird das angenaherte Gesetz
bei der Ermittelung zu Grunde gelegt, dass die Massen den
Hohen proportional sind. Es besteht also die Proportionalitat :

m:h=mz th,

worin m und A aus der Tafel, 1, aber durch Messung bekannt ist;
z. B. ein Bestand habe 81,27 qm Grundflache im 65 Jahre, seine
Hohe sei 23 m: Er gehodrt demnach in die Bonitat II. Die
gesuchte Masse ist daher:

1035 : 23,93=x : 23 mx =994,78 {m.
(m:h ist der Factor zur Hohe, den wir auf pag 364 berechnet
finden. Entnehmen wir dort denselben, so haben wir h nur noch
mit demselben zu multipliciren, um die Masse zu finden).

d. Wenn ein Bestand im Alter und in der Hohe, jedoch
nicht in der Grundflache gleich ist, so kann man die Masse nach
dem Verhdltnisse der Grundflache berechnen. Es istm: g=mzx: gy
in welcher Proportionalitat wieder m und g aus der Tafel, g,
aber aus dem Bestande bestimmt werden muss. m: g ist aber
die in der Ertragstafel berechnete Richthoéhe; wir brauchen dem-
nach nur noch diese mit der Bestandsgrundflache zu multiplicir-
en, um den Ertrag zu erhalten; z. B. Ein 65 jahriger Bestand,
welcher der Hohe nach genau II Bonitaét angehort, habe nur
80 qm Grundflache, dann ist Masse=1016 fm, da die Richt-
héhe=12,7 m und das Produkt 80X12,7=1016 ist.

e. Differiren in den unter c und d eben gedachten Fallen
auch die Alter gegen die Tafel, so miissen diejenigen Factoren
zur Hohe resp. Richthodhen genommen werden, die zu dem gefun-
denen Bestandsalter gehdren. Ist der Bestand z. B. nicht 65
jahrig, sondern 68 jahrig, so wird bei abweichender Hohe (von

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 359

23 m) diese mit 43,33 zu multipliciren, mithin die Masse=g96,59
fm. sein. Sie ist niedriger als vorhin, weil ein Bestand, der erst
im 68. Jahre die Hohe 23 m. erreicht, einer niedrigeren Bonitat
angehoren muss, als ein solcher, der schon im 65. Jahre diese
Hohe hat.

Weicht der 68 jahrige Bestand aber nicht in der Hohe von der
Tafel ab, sondern nur in der Grundflache und ist diese wie vorhin
=80 qm, so ist die Masse 80X13,0=1040 fm. Hat ein Be-
stand nur im Alter, aber auch in diesem nicht mit der Tafel
Uebereinstimung, so ist die Masse nach der Gleichung

Meh. f

zu berechnen, so dass nur f aus der Tafel entnommen werden
kann ; man bestimme diese Grosse jedoch nicht nach dem Alter,
sondern nach der Hohe, weil sie zumeist von dieser, vom Alter
hingegen nicht bemerkbar abhangt. Ist z.B. in einem 65 jahrigen
Bestande die Grundflache=80 qm, die Héhe=23 m, so ist die
Formzahl, wie aus der Tafel fiir Bonitaét II zu entnehmen=538
daher
M=80 X 23 X0,538=989,92 fm.

Einfacher ist es, wenn man f.h direct aus der Tafel entnimmt.
Zur Hohe 23 m. gehort dann bei Bonitat II die Richthohe 12,3 ;
mithin ist die Masse 80 X 12,3=984 fm.

6. Die vorstehenden Ertragstafeln niitzen aber in noch
hoherem Maase bei Massenzuwachsermittlung der Bestande.
Wenn man z. B. feststellen will, um wie viel Festmeter ein
normal bestockter 60 jahriger Sugibestand, welcher nach seiner
Hohe und Grundflache genau der II Bonitaét angehort, in den
nachsten 15 Jahren zuwiachst, so hat man nur aus der Ertrags-
tafel II. Bonitat die Holzmasse des 75 jahrigen Bestandes mit
r113-fm und die des 60 jahrigen Bestandes mit 983 fm. heraus-
zuschreiben, und erhalt in der Differenz beider Zahlen, namlich
in 1113-983=130 fm. den 15 jahrigen Haubarkeitszuwachs pro
Hektar. Fiele der fragliche Bestand, was in der Regel der Fall
sein wird, hinsichtlich seiner Standortsgiite zwischen zwei Boni-
taten hinein und ware auch seine Bestockung eine abnorme, so
miissten dann natiirlich dieselben Reduktionen wie im vorigen
Beispiel vorgenommen werden, um den konkreten Zuwachs zu
erhalten.

360 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

7. Unsere Ertragstafeln sind ferner auch néthig zur Ermitt-
lung des Normalvorrathes aller méglichen Umtriebszeiten. Will
man z. B. den Normalvorrath bei 100 jahriger Umtriebszeit fiir
120 Hektar haben, so braucht man nur die 100 Massenglieder der
betreffenden Bonitat zu addiren.

8. Endlich kénnen die Ertragstafeln noch zur Loésung aller
Fragen der Waldwerthberechnung und der forstlichen Statik
verwerthet werden. Man braucht nur die in denselben stehenden
Holzmassen in Sortimente zu zerlegen, letztere dann mit den
zugehorigen reinen durchschnittlichen Holzpreisen zu _ multi-
pliciren; dann erhalt man in den Summen der einzelnen
Produkte den in Geld ausgedriickten Werth des Holzes pro
Hektar fiir jedes Bestandsalter und auch fiir jede Bonitat.

ie cE Ee
Zuwachsgesetz von Sugibestanden.

mit besonderer Beriicksichtigung der Weber’schen

Zuwachsgesetze.

I. Holzmasse.

a. Fahrlicher Massenzuwachs.

Ich habe, zum Vergleich der aus meinen Untersuchungen
sich ergebenden Massencurve, auch die Curve in _ punktirten
Linien beigefiigt, wie sie sich aus der Formel

I
100 p3 ee
berechnet, welche mein verehrter Lehrer Prof. Dr. R. Weber
in Miinchen abgeleitet hat; in dieser Formel bedeutet die
Grundzahl, welche die Wachsthumsenergie angiebt, x das Alter
(Siehe Pl. XVIII et XIX).

Aus diesen Curvendarstellungen sehen wir, dass auch die

japanischen Sugibestande jenem Zuwachsgesetz folgen, welches

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 361

in europdischen Waldungen Geltung hat. Unsere Massencurven
ergeben folgende Angaben von p:

TAB. VI.
| Bonitat. Jugend-
stadium
| I II III IV im Jahre.
Schaftholz .. .. .. ..| 2,47-2,49| 2,35-2.37 | 2,24-2,25 | 2,09-2,10 15
Schaft- und Astholz .. ..| 2,50-2,52| 2,39-2,41 | 2,28-2,29 2,15 15

Zum Vergleiche nehmen wir Angaben iiber deutsche Fichten
und Weistannen aus Webder’s Forsteinrichtung heraus, weil beide
Holzarten im Zuwachsgang am meisten unseren Sugibestanden

ahnlich sind (Tab. VII).

TAB. VIT.
if II | Ill | 1V Vv Jugend-
Bonitaten. stadium
Werthe fiir der Kurven. Jahre.
Fichten.
Im Harz nach Hartig.. ..| 2,4-2,3 | 2,3-2,2 _— — — 40
In Wirthenberg nach v. Baur| ca.2,2 | ca.2,1 |1,96-1,90| 1,71 — 20
In Sachsen nach Kunze .. 2,3 ApuG | 2,03 1,89 — 15
In Norddeutschland nach
Schwabach.. .. -) o| 24-253, || Ca-2,2) | 2,1—2,0 | 1,9—-1,8 | ca.r,7 20
In Siddeutschland nach
Schwabach.. .. .. .«+| 2,4—2,3 | Ca.2,2 |2,13—-2,10| 2,0-1,9 | ca.1,8 ae

In Gouy. St. Petersburg nach

de Bedemmar .. .. «| 1,9-1,8 |1,80-1,74|1,64—-1,63| ca.1,5 | 1,4-1,3 25

Weisstannen.
25 u. 40
In Baden nach Schuberg ..| 2,4-2,3 |2,25-2,20|2,15-2,05] 2,0-1,9 | ca.1,9
In Wirttemberg nach Lorey.| 2,5-2,4 | ca.2,3 | ca.2,2 — — 50u. 60

Aus beiden Tabellen (VI, VII) sehen wir, dass das von
Schwabach gefundene p fiir Fichten am nachsten unserem Sugi-p
kommt, nur im Jugendstadium sind gréssere Unterschiede be-
merklich, weil unsere Sugz viel rascher wachst, als deutsche
Fichten.

362 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.
b. Laufender jahrlicher Massenzuwachs.

Es culminirt der laufende jahrliche Zuwachs :

bei Bonitét I um das 35 ste Jahr
” ” 100 330 335) 90 ”
” ” i, »» 35-49 5, ”
” ” IV ” ” 45» ”

Es geht aus diesen Zahlen hervor, dass beim Zuwachs an
Gesammtmasse dey Kulminationspunkt bet besseven Standortverhalt-
nissen frither eintritt als bet ungiinstigeren, wie sich auch aus
Weber’s Theorie) ergiebt.

Beim Schaftholz finden wir ebenfalls den obigen ahnliche
namlich :

fiir Bonitat I um das 35 Jahr

” ” II 99 99 35 ”
” 99 III ” 99 40 9°

” 9 IV 99 ” 45 29

Nun wollen wir zum Vergleich unsere Zahlen mit den
Zahlen fiir europaische Fichten und Weistannen in einer Tabelle
folgen lassen: (Tab. VIII).

LAB VIEL

Zeitpunkt und Massenbetrag der Kulmination des laufenden

Zuwachses.

I Bonitat. | II Bonitat.| III Bonitat. |[V Bonitat.] V. Bonitat.

Jahr.| cbm,] Jahr.] cbm.) Jahr. | cbm.| Jahr.| cbm.| Jahr.] cbm.

Fichte.

Nach v. Baur... .. ..|/27-30] 19,2 |]38-39| 13,0 |27-46 | 8,0/31-50) 6,0] — | —
Nach Schwabach
in Norddeutschland ../30-35/ 22,7} 40 |17,2| 55 | 13,2] 60 | 9,8] 65 | 7,5
in Siiddeutschland ..| 40 | 23,2] 45 | 16,6} 60 | 13,1] 85 |10,5] 80 | 8,0

Weisstanne.
Nach Schuberg .. ..| 25 | 22,4] 40 | 17,2 /35-40 | 13,5 40-50! 9,2 |50-80] 7,05
Nach Lorey .. .. ..| 80 | 16,0] 90 | 12,8 /95-105| 10,8} — —|—]—
Sugiin Japan ..| 35 | 26,8] 35 | 22,4 |35-40 | 18,2] 45 |14,8/ — | —

(1) Weber's Forsteinrichtung Seite 249.

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 363

Diese Tabelle zeigt, dass der Kulminationszeitpunkt von
Sugt dem der deutschen Fichte und Weisstanne ziemlich nahe
steht, dass aber der Massenbetrag der Sugz bei der Kulmination
bedeutend grosser ist.

c. Massendurchschutttszuwachs.

Bei der Gesammtmasse ist auch die Culmination des
Durchschnittszuwachses vorhanden: Sie tritt ein
fiir Bonitat I um das Jahr 45-50
55 65 1 an hee 50
» me LEE ee a a5
+ Vie te OOS
Die Zeiten liegen also bet geringeren Bomitdten immer 5 Fahre
spater. Fe geringer die Bonitat, wm so spater tritt die Culmination
ein und um so linger schtebt sich die Abnahme des Massendurch-
schnittzuwachses hinaus.
Beim Schaftholz beobachten wir Culmination
fiir Bonitat I um das Jahr 50
” ” II ” cy) ” 55
En 5 1 et i | pe eae)
4 ip TV "55° 821 1957 60-65
Auch hier gilt dasselbe Gesetz wie fiir die Gesammtmasse,
wahrend die absoluten Jahreszahlen immer um 5 grosser sind.
Da die Umtriebszeit fiir den gréssten Massenertrag im
Kulminationsjahre des Massendurchschnittszuwachses liegt, so
kann man sagen, dass bei unseren Sugzwaldungen der grésste
Massenertrags-Umtrieb ftir 1 Bonitat 50 Jahre betragt und fiir
je eine Bonitatsklasse schlechter um 5 Jahre zunimmt.

d. Verhdltniss von laufendem jahrlichen- und durchschntttlichen
Massenzuwachse.

Ein Blick auf Pl. XXVI lehrt uns, dass der laufende jahr-
liche Zuwachs sehr schnell bis zu einem Maximum ansteigt, um
von da ab wieder schnell herab zu sinken, wahrend der Durch-
schnitiszuwachs viel langsamer steigt und fallt, als jener; der Cul-
minationspunkt liegt bei ersterem viel hoher, als bei letzterem.

Ausserdem sieht man, dass der Culminationspunkt des
Durchschnittszuwachses immer derjenige Punkt ist, wo dessen
Curve mit der Curve des laufenden jahrlichen Zuwachses kreuzt.

364 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Prof. Weber hat fiir den Durchschnittszuwachs die Formel

100p>(I 55x)

itn
abgeleitet. Nach dieser Formel habe ich ebenfalls die Curve des
Durchschnittszuwachses gezeichnet. Wie aus der punktirten
Linie in P]. XXVI zu ersehen, fallt diese mit dem beobachteten
Durchschnittszuwachs fiir unsere Sug? annahernd zusammen.

D+x«=

e. Verhaltniss von Massen- und Hohenzuwachs.

Aus diesen Verhaltnissen habe ich den Factor zur Hohe
berechnet und die Zahlen in Tabelle IX zusammen gestellt.

(TAB. SLX.

Factoren zur Hohe in qm.
Alter.
Bonitat I. Bonitat II. Bonitat III. Bonitat IV.

5 12,50 TL, Ox 9,47 6,35
IO 23,18 21,65 19,08 13,29
15 23,87 22,42 20,17 15,70
20 25337 24,40 22,51 19,07
25 28,17 27,13 25,38 22 oe
30 31,62 30,37 28,58 25537
35 3517 33,77 31,84 28,56
40 38,16 36,70 34,68 31,46
45 40,56 39,13 3718 34,09
50 42,43 41,03 39,16 36,32
55 43,53 42,31 40,57 38,07
60 44,02 42,91 41,39 39,24
65 44,26 43,25 41,93 40,04
70 44,26 43,38 42,19 40,52
m5 44,18 43,43 42,32 40,86
80 44,07 4331 42,31 40,93
85 43.93 43,17 42,21 40,85
go 43,72 43,02 42,02 40,09
95 43,62 | 42,83 41,85 40,51
LOO 43535 42,59 41,65 40,32

ST

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 365

Hieraus erfahren wir, dass die Hohenzunahme so ziemlich
denselben Gesetzen gehorcht, wie die Massenzunahme, dass sich
jedoch kleine Unterschiede zeigen. Es besteht nur aproximativ
die Proportionalitat der Massen mit den Hohen; auch ist der
Factor zur Hohe fiir ein und dieselbe Tafel nicht immer constant,
sondern verdndetlich, was Herr Wetse zuerst in seinen Kiefer-
ertragstafeln erwahnt hat. ;

II. Bestandsmittelhéhen.

Prof. Dr. Weber hat die Hohe h, bei dem Alter a mit dem
Grenzwerth h,,,, algebraisch durch die Formel :

ha = Vind ge) verbunden.

Nach dieser Formel habe ich die Resultate meiner Unter-
suchungen tiber die Bestandsmittelhohe berechnet und zur gra-
phischen Darstellung gebracht. Die stark gezogenen Linien in
Pl. XX zeigen diesen Wachsthumsgang, daneben wurden die
Kurven punktirt aufgetragen, welche man bekommt, wenn man
in der Formel den bestimmten Grenzwerth von Aya, mit 35 m
einsetzt.

Diese Ergebnisse zeigen deutlich dass der Zuwachsgang
der Bestandsmittelhohe bei der Suwgz auch dem der deutschen
Holzarten™ sehr ahnlich ist und dass dze Hohenwachsthumsenergte
mit zunehmender Bodengiite grosser wird, (siehe Tabelle X).

TAB. X.

I Bonitat. |II Bonitat. |III Bonitat./|[V Bonitat.| V Bonitat.

P der Bestandmittelhohe.

Sugi in Japan Sey eke 2575 1,8-2,0 I,3-1,5 I,I-1,2 _—

Kiefer, Fichte, Buche und

Tanne in Deutschland. 2-255 1,5-2 I I-0,7 055

(1) Weber's Forsteinrichtung Seite 157.

366 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Die absolute Groésse von # ist fiir Sugi etwas groésser, als fiir
deutsche Holzer; wahrend des Jugendstadiums von Sugi aber
ist sie viel geringer als fiir deutsche Hélzer (mit Ausnahme
der Kiefer): Kiefer’ 5) Jahre

Fichte 10 5

Buche 15 %5
Tanne 15-25 ,,
Sugt 5 y

Der laufende Ho6henzuwachs culminirt bei

Bonitat I mit 11,35 m zum 20 Jahre (die Masse 35 Jahre)
9 II J) 9,14 oe) 99 20 99 ( 29 ” 35 9 )
” III b) 6,93 ” ”» 20 ”? ( ? 29 35-49 29 )
9 IV ” 6,44 ” ” 25 ” ( ” oe) 45 ” )

Es zeigt diese Tabelle. dass der Héhenzuwachs der Sugt
‘betrachtlich frither kulminirt als der Massenzuwachs, und zwar auf
besseren Standorten etwas friiher als auf geringeren.

Die durchschnittliche jahrliche Zunahme der Hohe culminirt
bei Bonitat I und II um das Jahr 25, wahrend bei schlechten
Bonitaéten etwas spater, namlich bei III um das 30, bei IV um
das 40.

Um Wachsthumsgange des Laufend- und Durchschnitts-
Zuwachses der BestandsmittelhGhe zu veranschaulichen, habe
ich sie in Pl. XXV. graphisch zusammen gestellt.

III. Stammzahlen.

Wir finden hier zunachst, dass die Stammzahl mit steigen-
der Bodengiite in gleichem Alter fallt: ;
Jahr 20 30 40 50 60 70 80 go 100
Bonitat I 3068 2311 1824 1441 1144 932 799 724 690
> II 3646 2826 2244 1798 1457 +1208 1040 938 885
* Ill 4224 334% 2663 2154 770 1483 1282 1151 1081
IV 4802 3856 3083 2511 2083 1759 1523 13605 1276
Nun vergleichen wir unsere Stammzahl von Sugz mit

europdischen Holzarten:

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 367

TAB; XI.

Die Stammzahl des Hauptbestandes auf 1 ha normaler

Waldflache :

Bonitat.

Holzarten, Wuchsgebiete.

Sugiin Japan ..

Fichte.
Wirttemberg nach v. Baur
Sachsen nach Kunze..

Harz nach R. Hartig

Mitteldeutsches Gebirge und Nord-

deutschland nach Schwabach

Siiddeutschland n. Schwabach..

Gouvernement St. Petersburg nach

Wargas de Bedemmar ..
Weisstanne.
Wirttemberg nach Lorey Se
Baden nach Schuberg ..
Kiefer.

Preussen, Bayern und Sachsen nach

Weise ates Vas
Hessische Main-Rhein-Ebene
Schwabach

Hessisches Buntstein-Gebiet

nach

| I | II jn

Beim 40 jabrigen Alter.

- +|1824|224412663/3083

. |2800)3 370|481016760

-|2460/3080)3530/4520

- +[3220]5700, —

+|3053|3947|5080/6543

+ -/1816)25 58/305 4/3909

+/2380)3130/35005070

Schwabach -/2490|3130 3630/3970
Norddeutsche Tiefebene nach

Schwabach -|I740|2370|3070)3980
Pommern uach R. Hartig --|r566] — | — | —

Wirttemberg nach Sfeidel

Gouvernement St. Petersburg nach

Wargas de Bedemmar ..

+ -|2050/2770|1430| —

- - [3060/3520 3980\4590

.|2380 4070|6030|79 1011000

9800)

Shia

775
62

H

4535 461

799

5280) 624 | 799

IV | Vv | I ) 1) ma] rv | Vv
Beim 100 jahrigen Alter.

6g90| 885 |1081|1276| —

950)1250)1600

9351200

886/1050)1370

TLOO h——

759% 942

568) —

638} 815|1070

go8/1115/1420

368 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Aus dieser Tabelle ersehen wir, dass unsere Stammzahl von
Sugt beim too jahrigen Alter immer grosser ist, als die von euro-
paischen Holzarten, wahrend bei 40 jahrigem Alter im allge-
meinen diejenige fiir Suwgi kleiner ist als die der europaischen
Baume; d. f. die Stammzahl fiir Sugi vermindert sich langsamer,
als die von europaischen Holzarten. Diese Thatsache versteht
man indessen leicht, wenn man sich das japanische giinstige
warm-feuchte Klima vergegenwartigt.

Betrachtet man den Verminderungsgang der Stammzahlen
ftir Sugi mittelst der Weberschen Formel fiir die Stammzahl-

verminderung :
I 0000
1,0 p*

so findet man, dass diese Formel fiir Sugz in Kzyosumt zu schnell

abnimmt und daher unbrauchbar ist. Wenn man aber statt
x

1,0 p* 1,0 p2 fiir Sugi benutzt, welcher Ausdruck nach Prof,
Weber erst nach Bestandsreinigung giiltig ist, so findet man
eine gute Uebereinstimmung. Auf Pl. XXII stellen die punkt-
irten Curven

6000

ropes
dar, da in unserem Sugibestand gewohnlich pro ha 6000 Stamme
gepflanzt werden. Es zeigt sich, dass die Stammzahlver-

minderung fiir Sug ungefahr der Formel
I

1,0p #
folgt und zwar liegt der Werth von p
fiir die Bonitat I zwischen 5,5-6,6
eens % II 5 ca. 5,0
Bey 85 34 III 5 ca. 4,0
sh tiat 9% IV 9 3,0-3,6

IV. Grundflachensummen.

Ein Blick auf Pl. XXI lehrt uns, dass die Grundflachen-
summe in dem ersten Dezennium sehr klein ist, dann aber rasch
ansteigt, um zwischen 40-60 Jahren einen Kulminationspunkt

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 369

zu erreichen, von dem an eine allmadlich immer langsamere
Zunahme stattfindet. Je besser dic Bodengiite ist, desto grdsser
wivd die Grundflachensumme, wahrend bet der Stammzahl das
Umgekehrte statt findet.

Die folgende Tabelle dient zum Vergleich.

DAB ci.

I Bonitat. |II Bonitat. |III Bonitat.|1V Bonitat.) V Bonitat.

Die grosste Grundflachensume im Bestande.

Sugi in Japan (beim 100
Valet) oe on “ed ibe 93531 85,03 76,76 68,48 —

Fichte(beim 100-120
\elre os Go al) | (lef) 56-61 52-53 43-46 36

| Weisstanne (beim)
PAGM alte) se) O7—-8x 59-67 53-59 48-55 —
Kiefer (beim 70-140 |
Jahre) .. .. «| 45-53 41-52 36-41 32-33 25-29

In Deutschland

Hieraus ersieht man deutlich, dass die Grundflachensumme
der Sug immer grosser ist, als die von deutschen Holzarten.
Die Sugi besitzen im grossen Durchschnitt 1,5 mal mehr
Grundflachensumme als die deutschen Fichten und Weisstannen.

Vergleichen wir unsere Resultate mit den Weber’schen

Zuwachscurven, deren Gleichung
2

G=t#

a. 1,0 p*

ist, wo G die Stammgrundflachensumme pro. ha. 1,0 * den
Nenner der Stammzahlformel bedeutet. Da wie wir gefunden

x

haben fiir Swgz immer 1,02 besser passt, als 1,0 f*, so benititz-
ten wir hier wieder folgende Formel

2G= jd
Oa
Nach dieser Formel construirte ich die Curven auf Pl. XXI, wo
ich die direct gefundene Grundflachensumme fiir Sugi zum
Vergleich beifiigte.
Man sieht auch hier, dass die Weber’sche, fiir Sugt durch
mich modificirte Formel durch die Beobachtung bestatigt wird ;

ob aber diese Modification fiir Sugi allein néthig ist, oder auch

370 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

fiir andere japanische Waldbaume, das ist eine eben so wichtige
wie schwierige Frage, die nur dann gelost werden kann, wenn
wir gute Ertragstafeln fiir andere Waldbaume Japans besitzen
werden. Abgesehen von diesen kleinen Abweichungen, die ohne
Zweifel durch die klimatischen Verschiedenheit bewirkt worden
sind, kinnen wir das Resultat unserer Untersuchung iiber die
Sugibestande in Kiyosumt dahin aussprechen, dass sie bestatigt
haben, was Prof. Weber in seiner “ Forsteinrichtung © behauptet
hat :

“Je grdésser die durch # ausgedriickte Wuchskraft eines
Bestandes ist, desto rascher nimmt zwar die Grundflache des
Einzelstammes zu, aber desto schneller sinkt auch die Stammin-
dividuenzahl und zwar erfolgt ersteres nach einer Multiplen-
reihe der Quadrate von #, letzteres nach dem umgekehrten
Werthe einer Exponentialreihe mit der Grundzahl t1.of.
Stammzahl und Stammgrundflache stehen demnach in einem
durch diese mathematischen Beziehungen ausgedriickten ver-
kehrten Verhaltnisse.”’

V. Durchmesser des Bestandsmittelstammes.

Bei einem Blick auf Pl. XXIII finden wir, dass bei gleichem
Alter der Durchmesser mit dem Sinken der Bonitat abnimmt ;
ferner, dass der Zuwachs des mittleren Durchmessers auch mit
der Weber’schen Formel D= 4% stimmt wie aus den punk-
tierten Linien ersichtlich wird.

In Folgendem ist ein aus P]. XXIII entnommenes # mit
dem der europaischen Holzarten zusammengestellt.

I II III IV Vv

Der Sugi in Kiyosumi yet: es Ao 1,2 0.9 0,7 a
Der Weisstanne mittleren Sutures: ades

Ti MSLAUDETES 2. (oes han ea 2 1,6 122, 0,9 0,6
Der Fichten in Norddeutschland n.

Schwappach .. .. «. : 2 1,4 0,9 0,6 0,4
Der Kiefern in Norddeutschland n.

wchmappach (5. 3) =. ees 5307 reas 0,8 0,6 0,3
Der Kiefern auf Bundsandstein in Hessen

N.dems, #56 tech (sk) want SS eee 0.9 0,6 0,4 _

Der Kiefern im Gouvernement St. Peters-
burg nach Wargas de Bedemmar .. 1,0 0,7 0,5 0,4 0,3

HONDA : ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. Sips

Hieraus ersehen wir, dass die Durchmesserzuwachsenergie
des Mittelstammes fiir unseren Sugi geringer ist, als diejenige
der europdischen’ Tanne und Fichte, wahrend die Massen-
zuwachsenergie der Swgi viel grosser ist, als die der letzteren ;
vielleicht eine Folge des Schlussgrades resp. der Durch-
forstungsverhaltnisse, ein Frage, welche ich spater zu erledigen
hoffe.

VI. Die Bestandsformzahl.

Ich habe in Pl. XXIV die Bestandsformzahlen nach dem
Alter graphisch dargestellt. Hieraus ergibt sich, dass bei
gleichem Alter fiir bessere Bonitat die Bestandsformzahl fiir die
Schaft-, so wie auch die fiir Ast- und Schaftholz immer geringer
wird, und dass beide Bestandsformzahlen anfangs sehr gross
sind, um dann zunachst rasch bis auf 0,5-0,6 spdter aber weit
langsamer zu sinken. d. h: Die Bestandschaftformzahl ist im
20 jahrigen Alter bei I Bonitat 0,504

We S573
III a 0,645
IV 98 0,671
wahrend im Alter von 30-100 Jahren
bei I Bonitat nur von 0,447 bis 0,439

9 II ”9 ” ” 0,497 ee) 0,467
9 eter ” ” ” 0,552 ” 0,497
” IV a) ” ”? 0,595 9 0,526

Aehnlich gestalten sich auch die Bestandsbaumformzahlen.
Ast und Schaft), ndmlich im

25 jahrigen Alter I Bonitat 0,573
II a 0,651
III A: 0,747
1 er 0,845

wahrend im Alter zwischen 35-100 Jahren
bei I Bonitat von 0,527 bis 0,464
Pa! | gee Pe 3 0,509 Gi ©1500
2 port 8 11 00524; 05543
al Se Os 7203 0,508

372 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

Bemerkenswerth ist es hier, dass bei den Bestandsschaft-
formzahlen besonders fiir I Bonitat im 30 jahrigem Alter einmal
diese zu einem ersten Minimum sinken, dann wieder bis zum
50 Jahre steigen, um von da ab sehr allmalich wieder zu sinken;
jenes Minimum ist aber bei schlechteren Bonitaten nur schwach
bemerkbar. Eine diesbeziigliche Beobachtung habe ich friher
in Deutschland gemacht und hiefiir eine mathematische Begriin-
dung geliefert. (Siehe: Eine Untersuchungsreihe tiber den Ein-
fluss der Hohenlage der Gebirge auf die Veraénderung des Zu-
wachses der Waldbaéume von S. Honda in der Allgemeinen
Forst- und Jagdzeitung 1892).

Der besseren Vergleichbarkeit halber sind nachstehend die
Formzahlen der einzelnen Bonitaten nach Altern zusammenge-
stellt.

(eB. SSIs

Die Bestandsschaftformzahl betragt :

Im Alter. Bei Bon. I. Bei Bon. II. Bei Bon. III. | Bei Bon. IV.
BK) 0,992 —= ——
15 0,628 0,714 0,806 0,794
20 0,504 0,573 0,645 0,671
25 0,461 0,517 0,584 0,620
3° 0,447 0,497 0,552 0,595
35 0,446 0,492 0,541 0,581
40 0,447 0,488 0,533 0,571
45 0,447 0,488 0,529 0,562
50 0,449 0,485 0,523 0,557
55 0,449 0,484 0,521 0,554
60 0,447 0,481 0,517 0,550
65 0,446 0,479 0,513 0,546
TO 0,445 Oar, 0,510 O34
TS 0,444 Dash) 0,507 0,539
80 0,443 0,473 0,505 0,536
85 0,442 0,471 0.502 0,532
go 0,441 0.470 0,500 0,529
95 0,440 0,468 0,498 0,527

100 0,439 0,467 0,497 0,526

a a RS SS

HONDA : ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 373

GAB: XV:

Die Bestandsbaumformzahl betragt :

Im Alter. Bei Bon. I. Bei Bon. II. Bei Bon. III. Bei Bon. IV.
se) 1,632 ===
15 0,887 1,033 1,219 P3807,
20 0,657 0,759 0,884 1,005
25 0,573 0,651 0,747 0,845
30 0,539 0,605 0,084 0,763
35 O527, 0,585 0,652 0,716
40 0,520 0,571 0,630 0,685
45 0,515 0,505 0,616 0,662
50 0,511 0,554 0,603 0,648
55 0,505 0,547 0,594 0,639
60 0,498 0,539 0,585 0,632
65 0,492 0,532 0,577 0,626
70 0,486 0,526 0,571 0,619
75 0,481 0,521 0,505 0,614
80 0,478 0,516 0,560 0,608
85 0,474 0,513 0,555 0,601
go 0,468 0,508 0,549 0,596
95 0,467 0,504 0,546 0,592

100 0,464 0,500 0,543 0,588

Nun betrachten wir die Bestandsschaftformzahlen nach der
Scheitelhohe :

TAB. XV.
Bei einer SeitelhGhe von I Bonitat. | Il Bonitat. | III Bonitat. | IV Bonitat.
5-10 m. 628 644 615 599
II-I5 m. 483 507 542 561
16-20 m. 447 489 523 541
21-25 m. 448 481 504 527
26-30 m. 446 471 — _
31-35 m. 441 — oe =

Die Bestandsschaftformzahlen nehmen also fiir jede Bonitat
mit zunehmender Hohe ab und bei gleichen Hdéhen fiir schlech-
tere Bonitat zu (mit Ausnahme der Unregelmissigkeit unter 10
m. Hohe).

Die Bestandsbaumformzahlen verhalten sich ebenfalls nach
diesem Gesetz, wie aus Tab. 16 ersichtlich:

374 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

TAB. XVI.
Bei einer Seitelhohe von I Bonitat. | 11 Bonitat. | III Bonitat. | IV Bonitat.
5-10 m. 887 895 816 775
tis} Fe 615 628 655 659
16-20 m. 533 574 600 617
SUSE) tle 525 542 558 592
26-30 m. 492 510 — —
S1S35_ a70 i i. ag

VII. Bestandsrichthohen.

Der Uebersicht halber folgen nachstehend die Bestandsricht-
hohen der einzelnen Bonitaten zuerst nach verschiedenem Alter
(Tab. XVIT und XVIII). Die Bestandsrichthéhen sowohl an
Ast- und Schaftholz als auch an Schaftholz, beginnen wie man
daraus erschen wird, niedrig und steigen mit zunehmendem Alter
allmalich an.

TAB. XV

Die Bestandsrichthohe fiir Schaftholz betragt :

Im Alter. | Bei Bon. I. Bei Bon. II. | Bei Bon. III. | Bei Bon. IV.
15 4:9 4,4 3,8 255
20 597 5:2 455 352
25 6,6 ovr 553 4,0
30 7,0 6,9 6,1 4,8
35 8,5 78 6,9 5,6
40 953 8,6 7,6 6,3
45 10,1 953 8,3 7:0
50 10,8 10,0 8,9 7,60
55 11,4 TO, 5 955 8,1
60 11,8 Lil 10,0 Be
65 12,3 L55 10,4 9,1
70 127 TQ 10,8 955
75 13,0 L252 112 959
80 13,3 12,5 11,5 10,2
85 13,6 1p) TLGy. 10,4
go 13,8 8-10) se) Io,7
95 14,0 ee 1255 10,9

100 14,2 13,3 L238 II,O

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. 375

TAB Sv ILL.

Die Bestandsrichthohe ftir Schaft und Astholz betragt :

Im Alter. Bei Bon. I. | Bei Bon. II. | Bei Bon. III. | Bei Bon. IV.
15 6,9 6,4 597 453
20 745 6,9 6,1 ANT,
25 8,3 7> 6,8 504
30 QoI 8,4 75 6,2
35 10,0 953 8,3 6,9
40 10,8 10,1 g,0 7,0
45 11,6 10,8 957 8,2
50 1252 I1,4 10,3 8,8
55 12,8 II,g 10,8 9,4
60 1352 12,3 Ey 10,0
65 13,6 1257 L,7 10,5
70 13,8 4351 R2eT 10,9
75 14,1 13:4 12,4 II,3
80 14,4 £3,6 12,7 rate 8,
85 14.6 13,9 12,9 11,8
go 14,7 14,0 ee ag 12,0
95 | 14,9 14,2 1353 12,2

100 15,0 14,3 ras 12,4

VIII. Das Zuwachsprocent.

Der Verminderungsgang des Zuwachsprocentes ist in PI.
XXVII und XXVIII graphisch dargestellt ; man sieht, dass das
Zuwachs procent des Bestandes sehr gross anfangt, dann sehr
rasch und spater langsamer sinkt.

376 HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI.

IX. Der Normalvorrath.

In Pl. XXIX haben wir den Zuwachsgang des Normal-
vorrathes mit starkgezogenen Linien gezeichnet und daneben mit
punktirten Linien Zinseszinsreihen (1,0h” —1) zum Vergleich.

Es ergiebt sich hieraus, dass die einer jeden Standortklasse
entsprechenden Normalvorrathe mit der Lange der Umtriebszeit
50-70 Jahre lang nahezu wie Potenzreihen zunehmen, und dass
mit geringerer Bonitat p abnimmt.

Dieses f fiir Ast- und Schaftholz ist zum Vergleich mit dem
p von deutschen WaAldern in folgender Tabelle zusammengestellt:

AB. XX

Aut Gundider I Bon. | II Bon. | III Bon. | 1V Bon. | V Bon,
Ertragstafeln fur : Procent p fiir die Reihen 1,0p" —1,
Buchen nach F. v. Baur .. 4-355 357-333 3,3-3 2,9-2,7 2,4

Fichten nach demselben .. 5-455 4,5-4,0 4,9-3,5 353-3 al

Fichten nach Kunze... ..| 5,5-5 5-455 4,5-4 357-3 _
Kiefern nach Schwappach .| 4,5-4 4-355 355-3 353-259 2,5
Sugiin Japan .. .. ..| 6,8-6,0 6,0 595-530 4,6 _

Die Zuwachsenergie der Normalvorrathe fiir Sugz ist also
etwas grosser, als die fiir deutsche Holzarten.

X. Das Nutzungsprocent.

Die Gréssen der Nutzungsprocente sind in Pl. XXVII und
XXVIII graphisch dargestellt, woraus hervorgeht, dass die
Nutzungsprocente mit schlechterer Bonitat grosser werden und
dass sie mit steigendem Alter allmahlig abnehmen.

Um die absolute Grosse der Nutzungsprocente an der
ganzen Holzmasse mit derjenigen deutscher Holzarten zu ver-
gleichen, habe ich in Tab. XX die betreffenden Daten zusammen-
gestellt, woraus man erkennt, dass das Zuwachsprocent fiir
unseren Sugi kleiner als fiir deutsche Fichten und Weisstannen,
aber etwas groésser als fiir deutsche Kiefer ist.

HONDA: ERTRAGSTAFEL U. ZUWACHSGESETZ FUR SUGI. Si,

AATS POX

Umtriebsjahre.

Nutzungsprocent fiir die Gesammte Holzmasse.

I Bonitat.
Fichten nach R. Hartig ..| 21,3]14,2 | 11,3 | 7,98! 5,97] 4,81] 4,07] 3,48] 3,03
|
Weisstannen n. Schuberg..| 14,3 | 12,1 8,29] 5,52 | 4,10 | 3,24| 2,68] 2,30] 1,96
Kiefern nach Schwappach..| 10,5] 6,98) 5,11 | 3,96] 3,20] 2,65] 2,26] 1,95] 1,70
Suge inijapan) |. -. || 13551] 8,8 Gre AS 7a eS) (2504253) E50) II lo

II Bonitat.

Fichten nach R. Hartig ..| 16,7|19,1 | 12,6 | 7,96| 6,02] 4,72) 3,97] 3,44] 3,04

Weisstannen n. Schuberg..| 14,1 | 11,5 8,52] 5,91 | 4,39| 3,41] 2,84] 2,39] 2,06

Kiefern nach Schwappach..| 10,8| 7,10| 5,41 | 4,00] 3,30] 2,74] 2,32] 1,98] 1,74

Sugiin Japan’ 2. «. «.| 24,0] 9,0 G5 aso) 357) "259 | 254 230 | 257,
III Bonitat.

Weisstannen n. Schuberg..| 14,4 | 11,1 8,65 | 6,27| 4,70] 3,70| 3,03] 2,54/ 2,18
5»32| 4,00] 3,24] 2,69] 2,29] 1,98) 1,74

Sugiin Japan .. .. «| 14,5! 9,5 | Sifs} || See | Stele) I yee |] Be | esa alerts}

Kiefern nach Schwappach..| 10,9| 7,52

IV Bonitat.

Weisstannen n. Schuberg..| 14,1 | 10,8 | 8,6 6,75 | 5.05 4,00 | 3,28| 2,74| 2,34
5,66 | 4,39 | 3,51 | 2,85 | 2,39 | 2.04| 1,76

SIS S24 297 | 23307) ESD

Kiefern nach Schwappach..| 11,1 | 7,57

Sugo in japan .% «« =| 16,0) 1055) ||) 735) || 5,0

Ueber den Einfluss wechselnder Mengen von Kalk und Magnesia
auf die Entwicklung der Nadelbaume.

VON

Oscar Loew und Seiroku Honda.

Es ist seit lange anerkannt, dass Kalkboden fiir die Land-
wirthschaft einen vorztiglichen Boden abgibt, und auch im
Forstbetrieb weiss man ihn zu schatzen. Kiefern gedeihen
besonders gut auf demselben. Der Kalk ist stets von mehr
oder weniger Magnesia begleitet, ein weiterer wichtiger
Nahrstoff fiir die Pflanzen, welcher aber unter gewissen
Bedingungen auch schadlich wirken kann, namlich wenn
seine Menge die des Kalks bedeutend tiberwiegt. Bei Experi-
menten mit Ndahrlosungen ladsst sich das leicht beobachten,
besonders wenn der Kalk ganz eliminirt wird. Aber dieser
schadliche Einfluss wird weit langsamer sich bemerklich
machen, wenn wie im Boden die Magnesia als schwerlésliches
Carbonat vorhanden ist. Nichts destoweniger ist auch hier
ein Unfruchtbarwerden durch zu hohen Magnesiagehalt
beobachtet worden. So lange die Menge der Magnesia
geringer ist als die des Kalks ist diese Gefahr wohl aus-
geschlossen wie z. B. beim Dolomitboden, wie Kellner mit-
theilt@) und Voelkey in England’ sowie Muntz und Girard in
Frankreich’) berichten.

Da uns die Frage interessirte, bis zu welchem Grade die
Entwicklung der Nadelbaume eine Storung durch steigende
Mengen von Magnesia im Boden erfahren konnen, stellten wir
einen Versuch mit jungen Pflanzen von Cryptomeria japonica,
Thuja obtusa und Pinus densiflora, den drei wichtigsten Wald-

(1) Wolff, Landw. Versuchs-Stationen 6,218; Raumcery und Kellermann, Ibid.
25,31; O. Loew, Ibid. 41,469.

(2) Adolf Mayer's Vorlesungen uber Agriculturchemie II, S. rrr (3. Aufl.)

(3) Sachsische Landw. Zeitschrift 1895, Nr. 24.

(4) Griffiths, Treatise on Manures 1889, S. 235.

(5) Les Engrais III, S. 333.

LOEW U. HONDA: EIJNFLUSS VON KALK U. MAGNESIA. 379

baumen Japans, an. Die Pflanzen wurden am 4. Mai 1894
in 5 Topfe verpflanzt (je zwei Stiick). Jeder Topf enthielt 5
Kilo Quarzsand, welcher vorher, um alle leicht ldslichen
Mineralsalze zu entfernen, mit concentrirter Salzsdure zwei
Tage lang unter 6fterem Umriihren stehen blieb, was nochmals
mit frischer Salzsdure wiederholt wurde. Dann wusch man
mit destillirtem Wasser, bis sich keine saure Reaction auf
Lakmus mehr zeigte.

Die jungen Pflanzen wurden sadmmtlich von Zeit zu Zeit
mit derselben Hauptloesung begossen, welche pro 100 cc.
enthielt :

Dikaliumphosphat 1g

Chlorkalium Ig
Ammoniumsulfat 2¢g
Eisenvitriol 6,18.

Jeder Topf erhielt von Zeit zu Zeit 50 cc. dieser Loesung,'’)
was vom 5. Mai bis 23. December 1894 34 mal und von 2.
Februar bis 28 September 1895 35 mal stattfand. Ausser
dieser Hauptloesung wurden noch zwei specielle Loesungen
hergestellt, deren eine rx procent Calciumnitrat und die andere
I procent krystallisirtes Magnesiumsulfat enthielt. Von diesen
Loesungen erhielten nach jedesmaligem Begiessen mit jener
Hauptloesung :

Topf Nr. I II III IV V.
Calciumnitrat-Loesung(2) 50 45 25 10 = we,
Magnesiumulfat-Loesung -— 5 25 40 SOMeCGs

Nach einem Jahre war ein ganz erheblicher Unterschied
bei den Pflanzen wahrzunehmen, der gegen den Herbst des

(1) Diese Salzmenge mag auf den ersten Anblick hoch bemessen sein, indessen
wenn man bedenkt, dass der grobe Sand keine Absorptionskraft besitzt, dass die
Pflanzen in der Zwischenzeit oft mit destillirtem Wasser—besonsers reichlich
wahrend der heissen Monate—begossen werden mussten und also die Salze hald in
die Tiefe des Topfes gefithrt wurden, wird man unser Verfahren gerechtfertigt
finden.

(2) Bei dem Ueberschuss an Stickstoff und Schwefel, der in Form von Am-
moniumsulfat in der Hauptloesung gegeben wurde, konnte sicherlich der Umstand
wenig mehr ins Gewicht fallen, dass das Calcium als Nitrat, des Magnesium aber
als Sulfat gegeben wurde. Beide wurden iiberdiess durch die vorher applicirte
Hauptloesung zum gréssten Theil in Phosphate verwandelt.—

380 LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA.

zweiten Jahres noch bedeutend zunahm. Am 5. October 1895
nahmen wir eine nahere Vergleichung der Unterschiede vor und
liessen eine photographische Aufnahme (PI. XXX) _herstellen.
Die Resultate sind in folgenden Tabellen angegeben.

TAB. I.—CRYPTOMERIA JAPONICA (SUGJ).

No. des Topfes. I II III IV Vv

Versuchspflanzen. a b a b a b a b a b

Lange der Keimpflanze..| 9,8] 9,0] 7,1| 6,0| 7,7] 7,5| 7,0] 7,5] 5.5] 85
Hohe im I Jahre .. .. | 11,8]12,5] 9,7] 9,0] 12,5|11,0| 10,0] 11,5] 8,7] 11,2

» soll 4, «2 se | 19,0)]'20,8)| 20,7 | 17,8 | 27,5 |. 17,5) | 22,0)| 1050 |)10,00 mee

Langster Zweig im I
Jahre... .2° ss ‘ss |/X8,0)|/ 225011145) | 10,35) 11,5 |) 750)|/£1,0)| TO;7i//2O, On mers

Langster Zweig im II
Jahre .. oe 2. «. |/£850)] 13,0] 18,4 |/7550 || 28,0)" 957)| 555211 1350/1, ol eee

Zahl der Zweige im I

HEIES 56 66 oo oe 3 5 3 | 4 4 4 4 4 3 3
Zahl der Zweige im I! |
VEIN sa) G5 05 oo) & 5 7 7 9 9 4 5 8 6
j
Gesammt Gewicht beider
Pflanzen in gr. im
frischen Zustande .. 28,2 44,0 40,1 26,5 22,7

Die Exemplare von Topf II und III waren am kraftigsten
entwickelt, die Zweige standen massig nach aufwarts gerichtet,
und die Nadeln waren von frischem Griin; wahrend in Topf
I (ohne Magnesia) und in Topf V (ohne Kalk) die jungen
Triebe schlecht entwickelt waren, die schwachen Zweige nach
abwarts hingen und die diinnen Nadeln eine gelbliche Farbung
hatten.

Ganz dhnliche Unterschiede ergaben sich bei Thya

obtusa :

(1) Ebermayer hat nachgewiesen (‘‘Lehre von der Waldstreu’’), dass zur Aus
bildung der Bldtter in einem Buchen-oder Fichtenhochwalde 5-6 mal mehr Mineral-
stoffe verwendet werden, als zur jahrlichen Holzproduction.

LOEW U. HONDA: EINFLUSS VON KALK

U. MAGNESIA.

TAB. II.—THUJA OBTUSA.

381

No. des Topfes. I II III 1V Vv
Versuchspflanzen. a b a a b a b

Lange der Keimpflanze.. | 5,0| 4,0] 5,5 Bol) SES] 2x0) VI OrGy) aS
Hohe im II Jahrecm. .. | 15,9] 11,5] 17,8 | 16,4 | 16,4 | 51,5 | 16,7 | 12,7 | 19,6 | 13,9
Langster Zweig im I

Jabrexenisy ec | rr b. i<j] 1050)||- 2501 || 12,0 10,6| 2,5|10,4|] 6,8] 10,5 | 10,0
Langster Zweig im 11

Jabretcmeyed 1 9,6| 7,3| 7,6 14,0) 7;0)|| 855) 7,3)||10;4 | 8,9
Zahl der Zweige im I

EWS co Gal Sa» Gayl 3 7 10 7 9 8 8
Zahl der Zweige im II

are we eh se) Yess) 3 7 8 10 6 5 5
Gesammt-Gewicht beider

Pflanzen in gr. im

frischen Zustande .. 15,6 20,8 19,7 16,4 15,8

|
TAB. III.—PINUS DENSIFLORA (MATSU).
No. des Topfes. IV Vv
Versuchspflanzen. a b a b

Lange der Keimpflanze.. 14,5| 74] — | 85
Hohe im J Jahre|-;. <-. 16,3| 8,6] — | 9,5

” ” II ” .- o. 2595 12,5 _ 18,0
Grésste Nadellange im

WES on “Goll oe 10,9| 6,9} — | 8,4
Grosste Nadellange im

Mp ahineceuers ef its = 10,0] 7,3} — Bai
Gewicht im frischen Zus-

Stand ems ero iets 10,6] 2,3| — | 1,6

In Topf Nr. III wurden hier leider durch Insektenfrass
beide Pflanzen und in Nr. V die gréssere der beiden Pflanzen
beschadigt und daher nicht weiter beriicksichtigt, Eine geringe
Beschadigung hierdurch erfuhr auch die kleinere Kiefer in Topf

382 LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA.

II, und die betreffenden Zahlen sind daher in den Tabellen
eingeklammert.

Gelbliche Farbung der Nadeln, war nur in TopfI und V
zu beobachten; auch waren in beiden diesen Fallen die Nadeln
am diinnsten, bei I zwar noch von betrachlicher Lange, bei
V jedoch auffallend klein, so dass der Kalkmangel hier in
auffalligster Weise im zweiten Jahre sich bemerkbar machte.

In folgenden Tabellen ist das Hdhenzuwachsprocent,
sowie die Zahl der Zweige und die Differenzen der gréssten
Nadellangen bei der Kiefer im ersten und zweiten Jahre
angegeben :

TAB. IV.—CRYPTOMERIA JAPONICA.

Nummer Hohenzuwachsprocent im Zweigzahl im
des Versuchspflanzen. I und II
Topfes. I Jahre. II Jahre. Jahre.
/

a 20,41 61,02 8

I b 38,89 66,40 be)

Durchschnitt 29,65 63,71 9

a 36,62 113,40 Io

I b 50,00 120,00 IL
Durchschnitt 43531 116,70 10,5

a 62,34 120,00 13

III b 46,67 77927 13

Durchschnitt 54251 98,64 13

a 42,86 120,00 8

IV b 5333 50,52 9
Durchschnitt 48,10 88,26 8,5

a 58,18 Q3,10 II

WY b 31,70 54240 9
Durchschnitt 44,07 73276 10,5

Hieraus ergiebt sich, dass im zweiten Versuchsjahre die
Abwesenheit des Kalks sowoh! als der Magnesia das Hohen-
zuwachsprocent sehr bedeutend herabsetzte. Die Zweigzahl
war bei Nr. III am grdéssten, wahrend bei V (ohne Kalk) zwar

LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA. 383

soviel Zweige wie bei II gebildet, aber doch nur dusserst kurz

waren, wie aus der Photographie ersichtlich ist. Siehe PI.
XXX.

TAB. V.—THUJA OBTUSA.

Hohenzuwachsprocent der Keim-

Nummer pflanze bis zum II Jahre. Zweigzahl im I und II Jahre.
des
Topfes. a b Durchschnitt a b Durchschnitt
I 218,09 187,50 202,75 II 14 12,5
II 223,603 248,93 236,28 18 15 16,5
Ill 228,00 228,57 228,29 16 14 15,0
IV 234,00 180,22 207,11 17 15 16,0
V 201,54 208,89 205,22 13 13 13,0

Auch aus diesen Daten folgt eine schlechte Entwickelung
bei I und V. Die Zweigzahl ist am groéssten bei II, III und
IV, also bei gleichzeitigem Vorhandensein von Kalk und
Magnesia.

TAB. VI.—PINUS DENSIFLORA.

Nummer Héhenzuwachsprocent im a Ae parame
des Jahre sind langer
Topfes. II Jahre, cpg :
grossere 64,24 0,70 cm.
I kleinere 43,16 1,40
Durchschnitt 53.70 1,05
grossere 84,57 3,70
II kleinere (16,92) 2,20
Durchschnitt (50,75) 2,95
grossere 56,44 0,90
IV kleinere 4534 0,40
Durchschnitt 50,89 0,65
grossere = =
Vv kleinere 36,84 —4,70 cm.

Durchschnitt — =e

384 LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA.

Das Hohenzuwachsprocent blieb somit hier im zweiten
Versuchsjahre bei Kalkmangel (V) bedeutend hinter den andern
Fallen zuriick. Interessant ist die bedeutende Langenzunahme
der Nadeln in Topf II, sowie die betrachtliche Langenabahme in
Topf Vim zweiten Jahre. Man wird auch an der Photographie
(Pl. XXX Reihe A) die Abhdngigheit der Nadellinge von der
Kalkzufuhr leicht erkennen konnen.

TAB. VII.—GEWICHT IN GR. IM FRISCHEN

ZUSTANDE.

Nummer des Topfes. \
Cryptomeria japonica.. 22,7
Thuja obtusa 15,8
Pinus densiflora gréssere . =

5 A kleinere . 1,6

Die Gewichte im frischen Zustande wurden bestimmt, nach-
dem man die Pflanzen aus dem gelockerten Kies vorsichtig
auszog, den Sand abwusch, und die Wurzeln anhaltend
zwischen Fliesspapier abtrocknete.

Bei Pinus war die Reihe wie schon oben erwahnt nicht
vollstandig; doch bei Cryptomeria und Thuja, ergiebt sich, dass
die Pflanzen der Topfe Nr. II und III am meisten zugenommen
hatten, also da, wo die Menge Magnesia die des Kalks noch
nicht iiberschritt, und dieser Umstand mag wohl bei Bestim-
mung der Bonitatsklassen der Bodenarten in Berticksichtigung
zu ziehen sein.

Die Unterschiede bei Cryptomeria sind bedeutender wie die
bei Thuja, was wohl dadurch erklart werden kann, dass jene weit
mehr Anspriiche an den Boden stellt als letztere, wesshalb sie
auch nur auf guten Boden iiberhaupt gezogen wird.

Noch méchte die Frage gestellt werden, warum in den
Topfen I und V, also bei Abwesenheit von Magnesia resp.
Abwesenheit von Kalk, im zweiten Jahre iiberhaupt noch eine
Entwickelung stattfinden konnte. Darauf mag geantwortet
werden, dass die Versuchspflanzchen aus einem guten Boden

LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA. 385

stammten und daher wohl von beiden Stoffen etwas auf-
gespeichert enthielten.“”

Berechnen wir aus den Mengen der angewandten Magnesia-
und Kalksalze die Mengen der freien Basen Magnesia und Kalk,
so ergiebt sich als Verhaltniss nahezu:

MgO : CaO
fic) Lopi > ii Ee 2 20
As ss III i 8 12
m9 56 IV I 0,5

Wir sehen somit aus den Ergebnissen des Topfes IV, dass
ein nachteiliger Effect producirt wird, wenn die Menge des
Kalks unter die der Magnesia sinkt,'?) wahrend anderseits ein
bedeutender Ueberschuss an Kalk wie bei Topf II nur giinstig
wirkte. Offenbar wird die fiir die Ernahrung notwendige
Magnesia auch dann noch mit Leichtigkeit von der Pflanze
aufgenommen, wenn sie in sehr geringer Menge vorhanden ist,
wie in Topf II.

Noch seien einige Bemerkungen betreffs der Reihe B der
beigefiigten photographischen Abbildung gemacht. In diesem
Falle waren sémmtliche Salze dem Sande direkt einverleibt
worden, und fand die Begiessung nur mit distillirtem Wasser
statt, und nicht mit Salzl6sungen. Hier konnten daher bei dem
mangelnden Absorptionsvermogen des Sandes die Bestandtheile
rasch in den Untergrund gefiihrt werden und daher eine
Aenderung der relativen Verhaltnisse in den oberen Schichten,
aus denen die Wurzeln vorzugsweise ihre mineralischen Baustoffe
entnahmen, stattfinden. Man erkennt zwar auch hier auf den
ersten Blick das Zuriickbleiben der Entwicklung bei Mangel an
Magnesia oder Kalk (siehe I und V); dass aber bei No. IV der
Unterschied von II und III weniger markant ist als in Reihe A
mag auf dem erwahnten Umstande beruhen.

Die wesentlichen Schliisse welche wir aus unseren Beobach-
tungen ziehen kénnen, scheinen uns folgende zu sein:

(t) Ueberdies ist die Entwickelung der Koniferen in den ersten Jahren ja weit
langsamer, als die der Laubholzer und vieler anderer Gewachse.

(2) Nach Rudolf Weber’s umfangreichen Untersuchungen enthalt sowohl
Kernholz als Splintholz 2-4 mal so-viel Kalk als Magnesia, und wird bei der Samen-
bildung die Magnesia aus dem Holz herangezogen.

386 LOEW U. HONDA: EINFLUSS VON KALK U. MAGNESIA.

ifs

iS}

Kalkboden ist auch dann noch als giinstig fiir Wald-
baume zu betrachten, wenn die Magnesiamenge relativ
sehr gering ist ;

Die Bonitat des Kalkbodens nimmt ab, wenn die Mag-
nesiamenge betrachtlich die Kalkmenge tiberwiegt ;

. Kalkmangel macht sich am auffalligsten bei der Kiefer

durch Production kiirzerer Nadeln bemerklich.

— EE

Ueber der Entstehung der Verkrumungen an Yotsuyamaruta.

(Sugi-Stangenholz)
VON

Dr. Seiroku Honda.

In der Umgebung von Tokyo besteht eine ausgedehnte Sugi
Stangenholzwirthschaft von Cryptomeria japonica, japanisch :
Yotsuyamaruta. Diese Bestande werden hier durch Pflanzungen
von circa 80-100 cm. hohen Pflanzen, 6000-8800 pro ha, herge-
stellt, welche meist einmal verschult und 3-4 Jahre alt sind.
Im fiinften Jahre nach der Pflanzung wird Beastung gemacht,
was je 2 Jabre spater starker wiederholt wird, um eine astreine
Stange zu erziehen.

Da dieses Stangenholz meist schon im Alter von 12-20
Jahren gehauen wird, ist es von Wichtigkeit gevadschaftige
Stamine zu produciren. Allein in der Praxis bekommt man
auffallend oft, durchschnittlich 60-70%, am unteren Ende
gekriimmte oder gedvehte Exemplare, so dass diese Kriimmung
oder Drehung die Verwendbarkeit der Stange im hohen Grade
beeintrachtigt. Dieser Theil ist ungefaéhr ebenso lang, als die
Pflanze bei der Umpflanzung gewesen war, nadmlich 60-120
em.) also im Durschnitt go cm. (Siehe Pl. XXXI).

Bei 16 jahriger Umtriebszeit liefert der Bestand pro ha.
im Durchschnitt 221 Festmeter Holzmasse von 5000 Stiick
Stammen. Da aber geradschaftiges Stangenholz mit 8 Meter
Lange 10 sen (ca. 22 Pf.) kostet, gekriimmte aber nur mit 7 sen
bezalt werden, so betragt der Verlust pro ha. 5000 x }X 3=100
yen (ca. 220 Mk.), somit sahrlich pro ha. 6,25 yen, und da das
Yotsuyamaruta-Gebiet um Tokye allein schon ca. 10.000 ha.
umfasst, entsteht im jahrlicher Verlust von 62.500 yen.

Nun umfassen die Suwgi-Waldungen im tibrigen Japan eine
Flache von etwa eine Million ha. Man kann danach den

(1) Die Pflanzung im Walde darf nicht friiher geschehen, weil kleinere Pflanzen,
wie sie in Deutschland verwendet werden, hier durch die wppig aufschiessenden
hohen Grasarten geschadigt wirden.

388 HONDA : SUGI-STANGENHOLZ.

enormen Verlust bemessen, wenn auch die vermehrte Nutzholz-
wirtschaft bei zunehmender Baumstarke den Verlust bei der
Stangenholzwirthschaft einigermassen aufhebt.

Es schien mir desshalb wichtig, der Ursache jener Ver-
kriimmung auf den Grund zu kommen und wenn méglich ein
Verfahren zur Vermeidung derselben aufzufinden, was wohl
nicht nur von wissenschaftlichem Interesse, sondern auch da
niitzlich sein diirfte, wo andere Holzarten zu geradeschaftigen
Stangen gezogen werden sollen.

Was die Ursachen dieser Verkriimmung anbelangt, kann
eben so wenig der Schnee als der Wind schuld sein; denn die
Verkriimmungen finden nach verschiedenen Richtungen statt,
fallen also mit einer bestimmten Windrichtung nicht zusammen.
Zudem zeigten sich jene Abnormitaten auch an solchen Oert-
lichkeiten, welche gegen jeden Wind vollig geschiitzt sind (Siehe
Pl. XXXI).

Es ist auffallend, dass tiberall da, wo der Sugiwald durch
Stecklinge erzeugt wurde oder durch wnattirliche Verpingung
entstanden ist, die Verkriimmung gar nicht auftritt, wie z. B.
auf der Insel Kiushiu, wo 50 cm. lange Aststangen 30 cm. tief in
den Boden eingesetzt zu -werden pflegen, oder in Akita-ken in
Nord-Japan, wo man Sugi-Walder mit natirlicher Verjiingung mit
Fehmelschlagbetrieb findet, welche sdmmtlich gerade Stamme
liefern, wahrend die dortigen kiinstlichen Pflanzungen wieder
die verkriimmten Stamme aufweisen.

Es lasst sich somit vermuthen, dass man die Ursache
jener Verkriimmung in der Pflanzung selbst zu suchen hat,
also die Orientirung des Stammes_ hierbei vielleicht die
Hauptrolle spielt. Um diese Vermuthung naher zu priifen,
habe ich mehrere Versuche angestellt, deren Resultate als
vorlaufige Mitteilung ich hiermit ver6ffentlichen mochte.

Flache A:

Eimjahrige, noch nicht verschulte Pflanzen von 10 cm. Hohe
wurden in 4 Reihen gepfianzt. Die erste Reihe enthielt Pflanzen
von derselben Orientirung des Stammes, in der die Pflanzen
gewachsen waren, so dass’die Siidseite des Stammes wieder
nach Siiden gerichtet wurde, wahrend die zweite Reihe solche
enthielt, welche in umgekehrter Stammorientirung gepflanzt

HONDA : SUGI-STANGENHOLZ. 389

wurden, so dass also die friihere Sitidseite jetzt mach Norden
gerichtet war. Inder dritten und vierten Reihe befanden sich
solche, die man um je go® gedreht ecinsetzte, so dass bei der
dritten die Siidseite nach Ost und bei der vierten nach West
gerichtet war. Die Entfernung der Reihen von einander betrug
20 cm. die Pflanzweite 18 cm.

Flache B:

Diese wurde mit dvewdahrigen, aber einmal im ersten Jahre
verschulten Pflanzen von go cm. Hohe in 8 Reihen zu je 20
Stiick bepflanzt und zwar wurden hier dieselben Anordnungen
wie oben eingehalten, wobei die ersten 2 Reihen die normal
orientirten Pflanzen enthielten.

Die Reihenentfernung betrug 95 cm. und die Pflanzweite
gO cm.

Die beiden Flachen A und B hatten tiberall gleiche Boden-
beschaffenheit und Standortverhaltnisse. Nach der Einpflanzung
habe ich nur Grdser ausjaten lassen und zwar jahrlich zweimal,
namlich im Friihsommer und Friihherbst. Sonst liess ich der
Pflanzung keine weitere Pflege angedeihen.

Resultate :

Auf der Flache A wuchsen die Pflanzen im ersten Jahre nach
der Pflanzung ungefaéhr 17,5 cm. im zweiten um 62,7 cm. im
dritten um git cm., die Hohe betrug also im Ganzen 149 cm.
Wahrend in der ersten Reihe fast alle Pflanzen gerade und nicht
drehend wuchsen, zeigten die der zweiten Reihe fast alle Bie-
gungen und Drehungen, als ob sie in ihre friihere Orientirung
zuriickstrebten. Einige wenige Pflanzen der ersten Reihe
zeigten zwar Biegungen, aber keine Drehung, was bei der
Bastbildung deutlich wahrgenommen werden konnte. Die
Pflanzen der dritten und vierten Reihe zeigten die namlichen
Verkriimmungen und Drehungen wie in der zweiten. PI.
XXXII enthalt die Photographie von einer Pflanze, welche
diese Erscheinungen sehr auffallig zeigt (der Finger zeigt die
Pflanzhohe).

Auf der Flache B wuchsen die Pflanzen in ersten Jahre nach
der Pflanzung ungefahr um 20 cm. im zweiten Jahre um 48 cm.

399 HONDA: SUGI-STANGENHOLZ.

im Dritten um 67 cm., es betrug also die gesammte Hohe 227
cm. Hier ergaben sich nun dieselben Wachsthumsverhdltnisse
wie bei A. Nur die Pflanzen mit beibehaltener Orientirung
wuchsen geradschaftig und insbesondere kein Drehwuchs machte
sich bei denselben bemerklich, wahrend die Pflanzen mit mcht
novmaley Orientirung wieder Drehung und Biegung zeigten wie
aus Pl. XXXII ersichtlich.

Meine Vermuthung, dass die Aenderung der Orientirung
des Stammes beim Umpflanzen die Ursache der Biegungen und
Drehungen sein kénnte, hat sich demnach in unserem Falle
bestatigt, und gibt ums die Vorschrift an die Hand, dass man,
um geradschaftige Stangen zu erziehen, bei der Umpflanzung und
in’s besondere bei der spateren Verschulung junge Pflanzen in
derselben Orientirung einzusetzen hat, in der sie gewachsen
waren.

Ich bemerke nur noch, dass man jetzt leicht eine Erklarung
fiir die bekannte Thatsache finden kann, dass Stecklingsbaume
immer geradschaftig wachsen, da hierbei die Lichtseite des
Astes immer nach Siiden gerichtet gepflanzt zu werden pflegt.

Nachdem diese Mitteilung bereits dem Druck itibergeben
war, fand ich in der diesjahrigen Augustnummer der forstlich-
naturwissenschaftlichen Zeitschrift, herausgegeben von Dr. von
Tubeuf, einen interessanten Artikel von Herrn Prof. R. Hartig,
welcher die anatomischen Verhaltnisse beim Drehwuchs der
Kiefer behandelt. Ich beabsichtige, bei Sugz ahnliche Unter-
suchungen anzustellen.

Besitzen die Kiefernadeln ein mehrjahriges Wachstum ?
VON
Dr. Seiroku Honda,

Wahrend G. Kraus die Ansicht vertrat, dass ein mehrjahriges
Wachstum der Kiefernadeln statt finde, nahm R. Meissner den
Standpunkt ein, dass zwar die Nadeln in verschiedenen Jahr-
gangen eine verschiedene Lange erreichen, sowie am Haupt-,
primdren, und secundaren Seitentrieb Unterschiede in der Lange
zeigen, aber im zweiten Jahre nicht mehr zunehmen.

Da ich mehrere Exemplare von Pinus longifolia zur Verfiigung
hatte, deren Nadeln die aussergewohnliche Lange von 50 cm. er-
reichen kénnen und also zur Entscheidung jener Frage besser ge-
eignet waren, als kleine Nadeln, nahm ich die Gelegenheit wahr,
wiederholte Messung an jener Species und des Vergleiches wegen
zugleich an Pinus Kovainsis und Pinus densiflora vorzunehmen ;
Die Ergebnisse sind in folgender Tabelle zusammengestellt :

Jahr der | Entwicke- | Baumteile der Lange der Nadeln, Differenz
Entwicke- lungszeit Entwickelu Mealy
lung. BR BSZEN: "8: | 12 Marz.| g Sept. | 2 Oct. | Marz-Oct.

I. Pinus Longiforia.

1894 |Spatsommer |Schaft(oberer Teil)| (14,20) 14,40 14,40 -+0,20
Ss Sommer »,(mittlerer ‘Teil)| 32,20 32,10 32,10 —0,10
of Frihjahr », (unterer Teil)| 45,70 abgefallen
- e Zweige 43,60 43,60 43,70 0,10

1893 i Schaft(oberer Teil)| 40,60 40,60 40,60 fo)

” oF ,, (mittlerer Teil) 40,00 39,90 39,90 —)55i(0)
” ” », (unterer Teil)} 43,10 43,10 43,20 +0,10

Il. Pinus Korainsis.

1894 Sommer Schaft 8,90 8,90 | 8,go fo)

55 4 Frihjahr - 12,60 12,60 12,60 fa)
1893 ” 45 7,10 7,00 7,00 =O.10
1892 ss Zweige 9,20 9,20 9,20 o

III. Pinus Densiflora.

1894 Spatsommer |Zweig(oberer Teil)| 10,60 10,70 10,70 +0,10
9 Sommer » (mittlerer Teil)} 12,00 I1,gO 11,90 —o,10
ri Frihjahr » (unterer Teil)| 12,00 12,05 12,05 +0,05

1893 Spatsommer Zweige 8,90 8,90 8,85 —0,05

1892 » ” 12,20 abgefallen —

KS

392 HONDA: KIEFERNADELN.

Da man beim Anblick eines vorjahrigen Triebes sofort
erkannte, dass die unteren Nadeln 2-3 mal langer sind, als die
oberen, spater entstandenen, so musste man selbstverstandlich
vermuthen, dass letztere im zweiten Jahre noch die Lange der
ersteren erreichen wiirden, aber gross war mein Erstaunen, bei
der Messung im zweiten Jahre zu finden, dass sie—mnicht mehr
zunahmen. Offenbar erreichen nur die im Friihjahre entstand-
enen Nadeln eine bedeutende Lange, nicht aber die spater
entstandenen; das Wachstum beider wird im Herbst abge-
schlossen. Wir miissen also wohl Meissney Recht geben, wenn er
das Wachstum der Nadel im zweiten Jahre bestreitet, mtissen
aber noch hinzufiigen, dass nicht nur die Linge an Haupt- und
Seitentrieb oder in verschiedenen Jahren wechselt, sondern auch
sich Unterschiede an derselben Axe eines Triebes ergeben: die
oberen bleiben kleiner als die unteren.™

eee eee SE Ee

(1) Die kleineren Nadeln, die im Spatjahr entwickelt werden, haben, wie ich
mehrmals beobachtete, eine kiirzere Lebensdauer, als die vollentwickelten ; erstere
fallen meist schon nach einem Jahre ab, wesshalb man an den Alteren Baumteilen
keine kleinen Nadeln mehr bemerkt, was zum Glauben verleiten koénnte, sie seien

noch gewachsen.

Bull. Agric. Coll. Vol. tl, Pl. XVIII

ate L {
—l |
7 rt imo | seal cae | Sa =
+ - ; ai + pet He: Be i at
itt if = eT a a a =o
—- == + | Se 4 | 4 | | ale +
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)\
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| | =2.5 APT
r Schaftholzmasse. [—~ F teh Ne
| elt aal| I. Bon.
r | t pol ese
| | = |e [ee We a ea | | | || | | oa om [oe ee |
nian 3B ieee
1200 + ce 5 — 24 -4--T |
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oc
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SAG TT

Lability and Energy in Relation to Protoplasm.
BY
Dr. O. Loew, Professor of Agricultural Chemistry.

I have explained in former Bulletins” that physiological
phenomena compel us to infer the existence of a /adz/e (active)
and a stable (passive) modification of albumin, and that the
former alone,—as such and in the form of active nuclein—, is
capable of leading by ‘‘ organization ” to living protoplasm, while
the latter, or stable, modification would correspond to the condi-
tion of the albumin in dead protoplasm and to the reserve-albumin
stored up in seeds and eggs. I have also described a highly
labile kind of albumin, which is stored up as reserve material
in the vacuoles and sometimes in the cytoplasm of living cells of
various plants,” and which undergoes a chemical change under
conditions that would kill living protoplasm.

I have further attempted to show that in regard to organic
compounds, lability implies for the atoms in labile positions a
distinct kind of energy consisting in continuous atomze motion
which increases under the influence of heat, molecular motion
passing here into atomic motion. Thus the cause of the respira-
tion of protoplasm, and the utilisation of the heat of respiration
for biological chemical functions of a katalytic nature, find a
simple explanation. My incomplete treatment of lability, how-
ever, has called forth critical remarks which induce me to offer
some further remarks. The critic says among other things :

“That the functions of living matter are explicable in mecha-
nical terms, physiologists of all schools alike admit. Dr. LoEw’s
merit lies in his offering us a plausible explanation of the mecha-
nical basis of wztal energy, of that activity in which, as is well
said by BUNGE (one of the foremost of the vitalistic school), ‘‘ lies
the riddle of life.”

(1) Bulletin, Vol. II, No. 2; No. 4.

(2) This remarkable substance was discovered by 7%. Bokorny and myself. Daiku-
hara made further communications on the extent of its distribution through the vege-
table kingdom (Vide Bulletin No. 2 and 4).

(3) Japan Mail, July 30 ; 1896,

3904 LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

“Let us turn to consider a difficulty in connection with Dr.
LoEW’s hypothesis—a difficulty involved in the whole conception
of /ability. The difficulty will be best explained by a concrete
example. ‘‘ One of the most interesting labile atomic groups,”

says Dr. LOEW, ‘‘ is the aldehyde group, —CXy in which the

oxygen exerts an attracting influence upon the hydrogen con-
nected with the carbon atom, this being generally tetravalent,
but sometimes only bivalent. The hydrogen atom is thus ever
oscillating between the carbon and the oxygen, as may be in-
dicated by the following formula :—

6) O

AGVa By Pin) Yes = C265
<r C=02H CoH C20=hH
(1) (2) (3) (4)

‘’ The hydrogen atom may be conceived as oscillating between
the carbon atom and the oxygen atom, as the contact-breaker of
a faradic battery oscillates between the electro-magnet and the
connecting point to the longer circuit. The defect in the analogy
we have chosen serves to point the difficulty of which we have
spoken. The oscillation of the contact-breaker depends on two
forces that alternately become effective in opposite directions, the
force of the electro-magnet overpowering the force of the spring,
and the force of the spring when the electro-magnet has ceased
to act. But can we find any similar play of alternating forces in
our hypothetically constructed labile compound? It does not
seem so. It is certain that the attractive force between two
chemical atoms increases enormously as the distance between
the atoms diminishes. Let us suppose our hydrogen atom to be
placed initially between the oxygen atom and the carbon atom in
such a manner that the pull is equal in both directions. If the
hydrogen atom now moves in the direction of, say, the carbon
atom, the attraction of the carbon atom at once becomes much

greater than that of the oxygen atom, and the-C¢} form should

be permanently assumed.

“Does, then, Dr. LOEW conceive that in a labile group
something takes place analogous to the changes in the faradic
battery when the contact-breaker is vibrating ? that the move-
ment—to return to our example—of the hydrogen atom towards
the carbon atom brings about a state of molecular forces within

LABILITY AND ENERGY IN RELATION TO PROTOPLASM. 395

the group whereby the opposite form of the group becomes pre-
ferable ; and conversely, that when the hydrogen atom has
moved back past the mean position and approximated itself to
the oxygen atom, the opposite phase now becomes superiorily
attractive ? That would seem to be implied, for in that way only
would be rendered possible the persistence in the group of an
oscillatory movement whereby kinetic energy is preserved in the
form of continuous atomic motion. But if that is Dr. LOEWw’s
view, he has not expressed it clearly; nor does he anywhere
indicate his conception of the mechanism by which this changing
play of the intramolecular forces may be supposed to be kept up.
To return to our original rough analogy, unless something can be
imagined more or less similar in its mode of working to the
alternate play of electric and mechanical stresses in the faradic
battery, the hypothesis of the oscillating atom in the labile group
seems difficult to maintain.”

I do not undervalue this criticism, for even specialists in
molecular science have not yet penetrated the mysteries of
lability of organic compounds. Thus, Grant Allen declares :
‘Atomic motion may be separative as in decomposition, or
ageregative as in the act of combining, the continuous or neutral
stage is not at present known though there is reason to think that
it exists.” ‘‘ Atomic motion is not known to have any continuous
form analogous to the orbital motion of a planet, the spinning of
a top, or the regular vibration of heat.” Ostwald argues:”
“Living organisms behave like katalytically acting substances.
The problem, however, of the mode of action of a katalytic sub-
stance is not yet solved. A start is made by the recognition
that hydrogen ions can, in proportion to their number, accelerate
certain reactions. It is physical chemistry to which we must
look to throw a light upon the riddle of life.’ In a later article,
however, Ostwald declares that katalytic actions could only be
expected to receive explanation by new principles which lay
beyond the laws of energy. Before I enter upon a further

(1) Force and Energy, Chapt. VI and VII.

(2) Chem. Centr.-B]. 1894 I. p. 4.

(3) This analogy was first recognised more than forty years ago by the physiologists
Ludwig and Lehmann, Cf, Lehmann, Lehrb. der physiolog. Chem, 1853.

(4) Aula 1895, Nr. 1. To be sure, nothing can be more hopeless than to search for
Ostwald’s “ Neue Principien, welche itber das Energiegesetz hinaus gehen |”

396 LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

discussion of the energy of aldehydes it may be worth while to
give a series of cases which show how much various atomic
groups and their relative positions can influence the stability of
compounds which ranges within very wide limits.” While, ég.,
certain substances easily resist higher temperatures, as benzene
and pyridine, others are decomposed by boiling water, as aceto-
acetic acid ; others again change even at low temperatures, as
cyanic acid or diamido-acetone.

Asa general rule it may be considered that the zxerease in
number of hydroxyl or amido-groups, as also the alternation of
positive and negative groups, depresses the degree of stability.
Glycerol is less stable then propyl alcohol; di- and triamido-
benzenes much less so than mono-amido-benzene ¢.e. aniline.
Diketohexamethylene is less stable than dioxyhexamethylene
(Baeyer), and acetoacetic acid less so than pyruvic or acetopro-
pionic (lzvulinic) acid ; but it becomes more stable by the intro-
duction of one more carboxylic group.

CH; CH,—COOH
Co Co
CH,—COOH CH,—COOH
Aceto-acetic acid | Acetone-dicarboxylic acid™
CHs, CH,
CO-COOH Co
ee i
CH, COO

er ere
Leevulinic acid

The increase in number of carboxylic groups considerably
diminishes the degree of stability in such cases in which two of
these groups are linked to the same carbon atom. With the

(1) In stable chemical compounds the atoms are placed in comparatively fixed posi-
tions relatively to each other and a change is not brought about except by relatively very
energetic actions.

(2) The ratio of positive to negative groups is in aceto-acetic acid 2: 2, while in
acetone-dicarboxylic acid it is 2:3. This difference may account for the greater stability
of the latter,

LABILITY AND ENERGY IN RELATION TO PROTOPLASM. 397

nitro-group, however, we observe a somewhat different behaviour
in regard to the influence of position ; thus I. 3 dinitropropane is
much less stable than the isomeric I. I or 2. 2 dinitropropanes.™
The stability of a compound against chemical attacks grows with
the increase of the number of the CH, or alkyl groups. Thus, form-
aldehyde enters much more easily into various reactions than its
homologues ; acetone dicarboxylic ether is no longer attacked by
phenylhydrazine after ¢wo methyl groups have been introduced
and it even resists the action of phosphorus pentachloride after
four methyl groups have entered.” Also the relative position
exerts an influence; thus, a methyl group in para-position in
diazobenzene leads to a greater stability than one in ortho-posi-
tion.® The 1. 3 keto-aldehydes of the form R—CO—CH,—CHO
are very unstable and undergo spontaneous condensation, while
after the introduction of a second alkylic radical in @—position,
a product R—CO—CHR’—CHO results that may even be dis-
tilled unchanged.” On the other hand we must not omit an excep-
tion observed with diazo-propionic ether, which is less stable
than diazo-acetic ether.”

CH—COOR CH;—C—COOH
a oN
N=N N=N
4 ~ (Ie
Diazo-acetic ether Diazo-propionic ether

The readiness with which an atom is replaced by another
atom or bya radical, may, ceterts paribus, be considered as a
measure of the lability of tts position. Thus, the hydrogen atoms
in a-position to a keto or carboxylic group show a higher
degree of lability than those in f or y-position,” but still more
labile are generally the hydrogen atoms of the amtdo-group.”

(1) V. Meyer, Ber. Chem. Ges. 25, 1709.

(2) Petrenko, Chemikerzeitung, 1895, p. 1880.

(3) “irsch, Ber. Chem, Ges, 24, 325.

(4) Claisen, Bayr, Akad. Ber. 1890 p. 447.

(5) Curtius, Ber. Chem, Ges. 28, 3037.

(6) Very numerous are the observations on varying lability of hydrogen atoms and
their substitutes in the benzene ring. Hydrogen atoms in a high degree of lability
are easily attracting atomospheric oxygen (autoxidations). By the vicinity of negative
groups as cyanogen, phenyl or carboxethyl, certain hydrogen atoms attached to
carbon can acquire in special cases the property of being easily replaced by metals; this
state, however, is not a genuine case of lability.

(7) Also the cyanogen group linked to nitrogen is more labile than when linked to
carbon (Ber. 28, 2074).

398 LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

But not only does the lability of these hydrogen atoms vary con-
siderably in different compounds, that of the entire amido-group
also does so, and the latter is, in some cases, easily split off as
ammonia, as in the case of diacetone-amine, or the pinylamine of
Wallach, and in other cases is readily replaced by hydroxyl, as
in the amides of acids. Of the y-amido-acids the amido-group
is also said to be easily removed with the formation of lac-

tones.

The relative posttion of the amido-group also exerts
a powerful influence upon the special character of a compound ;
thus ortho and para-amido phenol are stronger bases than aniline,
while meta-amidophenol is a weaker one, than aniline.

Of special interest is the ¢ransztion of a labile compound into a
stable modification. Incertain cases the latter can easily be re-
transformed into the former, in others it is difficult. Of nitro-para-
aceto-toluide a yellow and a colourless modification exist ; the
former is more stable at a high temperature than the latter but
can easily be retransformed into it. The oxime of oxynaph-
thoquinone-imide exists in two forms, a red one, stable in alkaline
solution; and a light yellow one, stable only in acid solution.

Claisen infers from his investigations on 1. 3 diketones that
some of them may exist in two modifications, viz., as
. C(OH)> G- CO... saad =,.CO.* CH: CON. Oi edepencdsmaeaeam
the nature of the attached radicals, the temperature, the nature of
the solvent, which of the two forms is the more stable under the
conditions present.” Two isomeric forms of the formyl-phenyl-
acetic ether have been isolated which can easily pass one into
the other, the liquid or a-modification corresponds to the enol
(hydroxyl-) structure, the solid or $-modification to the
aldo-structure.© The former which alone gives a blue violet

(1) Ber. Chem, Ges. 17, 203. The replacement of an alcoholic hydroxyl group
by an amido-group is only in certain cases easily accomplished, as in cyanhydrines, in
nitrosonaphthol (Ber. 1'7, 393) and in the ortho-nitro-derivative of the phenyl-B-
oxypropionic acid (Zizhorn, Ber. 16, pp. 2645 and 2651). Evidently the influence of
a negative group is required for enabling such an exchange.

(2) Lelimann and Gross, Ber. Chem. Ges. 24, R. 107.

(3) Gattermann, Ber, Chem, Ges. 28, 1733.

(4) Kellermann and Hertz, Ber. Chem. Ges. 29, 1145.

(5) Ibid. 29, R. 499.

(6) Wisticenus, Ibid. 29, R. 503. Brith/, Ibid. RK. 484. In this case the aldo-
modification has a decidedly acid character, while in the above mentioned case of
Claisen it is the enol-modification.

LABILITY AND ENERGY IN RELATION TO PROTOPLASM. 399

reaction with ferric chloride, is the more stable modification at
higher temperature and occupies a smaller molecular volume
(¥. Traube, 1896). In alcoholic solution the aldo-modification is
gradually produced while in chloroform the enol-modification is

preserved.
O— Cr HO—C-—H
CH . CsH; C : Gin
COOR . COOR .
Aldo-modification, Fnol-modification.

F. Traube also has shown the probability of the existence of
two isomeric forms of aceto-acetic ether, each of which very easily
can pass into the other.” Labile compounds, however, often
undergo such changes that reversion to the original substance
either requires the expenditure of a considerable amount of energy,
as in cases of polymerisation, or has become entirely impossible,
since the chemical structure has been altered too much, as, ¢.g.,
when amides are formed from oximes, or when ortho nitro-benzene
compounds change, by atomic migration, into amido-compounds,
the lateral chain exchanging a portion of its hydrogen against the
oxygen of the nitro-group.” Also the changes which amido-acetone
and amido-ethyl aldehyde undergo spontaneously under ordinary
conditions, when liberated from their compounds with acids,
belong to this group of phenomena.”

To give in all these cases a satisfactory explanation of the
degree of lability is not an easy matter, for want of clear con-
ceptions of chemical affinity and chemical energy. Ostwald

(1) Ber, Chem. Ges. 29, 1715. ‘The observations of Mried/ender, Gehring, Knori
and others had already shown that this ether can react in two distinct ways.

(2) The rule that oxygen connected with nitrogen in a compound can easily migrate
to carbon, but can never do so in the opposite direction, is easily explained on general
chemical principles.

(3) Of further exampies may be mentioned: By treatment with alkaline solutions
phenylazoxazol changes into the isomeric oxime of benzoyl cyanide (Russanow),; the
closely related ‘furazan carboxylic’ acid into cyan-nitroso-acetic acid (Vo/f and Gans;
Ber. 24), and a-methylisoxazol into cyan-acetone (C/aisen, ibid. 25). Isodiazo-naphthalin
isolated from its sodium combination changes at. once into ordinary diazonaphthalin
(Bamberger; ibid. 2'7), Phenyl hydroxylamine changes in contact with acids into-para-
amidophenol, and the=nitrosamine of that compound changes spontaneously after some
time (Bamberger; ibid. 27).

(4) Outlines of General Chemistry, p. 208.

4cO LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

writes: ‘‘ Chemical energy is to us the least known of all the
various forms of energy, as we can measure neither it nor any of
its factors directly. The only means of obtaining information
regarding it, is to transform it into another species of energy. It
passes most easily and completely into heat.” Later on™ he
declares: ‘‘ Chemical energy can be separated into two factors,
intensity and capacity. The doctrine of the intensity of chemical
energy embraces a large part of those phenomena that were
called affinity.”

I have adopted the definitions of Grant Allen (1. c.): ‘ Che-
mical affinity is the force that aggregates atoms, chemical energy
is motion which separates atoms and resists the aggregation of
atoms.” ‘‘ Force and Energy, the aggregative and the separative
powers are incessantly opposing and antagonising one another in
all bodies, great or small. The amount of aggregation reached
by any system at any point of time depends upon the relative
proportions of its forces and its energies at that moment.”

This holds good also for the complicated molecules of organic
chemistry. Ina very stable compound, the force of affinity be-
tween its atoms preponderates, and only a small amount of energy
is present.” On the other hand, substances that very easily
enter into reactions can hold their components only loosely bound,
and the force of affinity is here counteracted by chemical energy.
Atoms are in an unstable equilibrium when a minute amount of
work suffices to lead to an alteration in the system; such atoms
have chemical energy.

But we must carefully distinguish between several kinds of
znstability, There evidently exist substances, certain atoms of
which merely possess potential chemical energy relatively to the
other atoms in the same molecule, as in the case of oximes and
nitro-compounds where the oxygen linked to nitrogen possesses
‘“energy of position” with respect to the carbon atoms. In cer-
tain other cases, however, atoms may possess, with respect to
others in the same body, k¢vetic chemical energy, being in a
continuous vibrating motion. But doubtless, there are also

(1) Chem, Central-Bl. 1894. 1. p. 4.

(2) The compounds are, of course, here considered merely with regard to their
chemical structure, 7.2. to the relative position of their own atoms ; not in their relation
to other bodies. Everybody knows, e. g., that in relation to free oxygen a// organic sub-
stances contain potential energy.

LABILITY AND ENERGY IN RELATION TO PROTOPLASM. 401

compounds in which both modes of energy exist at the same
time.

Compounds in which much potential chemical energy is ac-
cumulated, capable of being suddenly liberated, are the explosives.
Such cases of instability have no particular interest in connection
with our special case. The instability I have in view, is the state
of lability caused by atoms kept in a fierce state of vibration,
which tend, therefore, to enter into numerous reactions. That such
labile compounds possess energy of the #zvedze kind, becomes
manifest from the fact that certain such compounds can, by
imparting their atomic motion to other distinct compounds, cause
certain chemical changes, without changing themselves in any
equivalent measure (katalysis). I have mentioned as examples
in a former Bulletin (No. 4) the katalytic action of maleic acid
upon ketazines, of ethaldehyde upon free cyanogen, of ethy] nitrite
upon thio-urea, and the actions of the enzymes.”

When a labile substance passes into a stable isomeric one,
its kinetic chemical energy either diminishes or wholly disappears,
and assumes, in equivalent ratio, the form of molecular motion,
z.¢., heat. Inthis transformation the molecular volume decreases,
while the melting and boiling points are raised. This is an in-
teresting fact, and in full accordance with Stohmann’s observa-
tions ; for the labile compounds have a greater thermic value than
the isomeric stable ones.”

Atomic motion of the hydrogen atoms in benzene has been
supposed by Keku/é to account for the existence of only one
modification of ortho-compounds, and by ZLaar™ for the hydrogen
atoms in tautomeric (absolute pseudomeric) compounds, to ex-

(1) We must always keep in mind that heat, z. ¢., molecular motion can under
certain conditions easily pass into atomic motion, and that the atmosphere is an immense
storehouse of heat energy ; thus the continuity of katalytic action is easily explained in
such cases where an exhaustion of energy would seemingly have to be expected.

(2) Journ f. prakt. Chem, 46, 530. ‘There seem to exist exceptions in which the
larger molecular volume is not coinciding with the larger thermic value, as eg, a
comparison of aldehydes with the isomeric alkylene oxides would indicate (Ber. Chem.
Ges, 24, 652). But we must keep in mind that we have in this case two compounds
of considerable difference in character before us, which stand to each other by no means

in such a relation as we have under consideration.

(3) The hypothesis of Zaaz has recently been declared improbable by Zraudbe (Ber.
Chem. Ges. 29, 1723), while Aeku/e’s view has,—at least to a certain extent—, been
replaced by that of Baeyer.

402 LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

plain their ability to react in two different ways; but both
these authors have failed to explain how the continuity of these
oscillations is kept up.

How, then, can the /adzity of aldehydes be explained by a
continuous atomic motion ?. The mechanism is in all probability
as follows: In the first place there is the hydrogen of the
aldehyde group attracted from two sides, that of the oxygen and
that of the carbon, the attraction of the former being, under the
given conditions, a little superior to that of the latter.

In the second place, the orygen atom moves closer to the
carbon atom as soon as the doubly linked state in the aldehyde
group(a) has passed into the simply linked state (b) of the hydroxyl

(1)
group.

This sudden motion of the oxygen atom influences of
course also the attached hydrogen atom, whose vibrations in
conjunction with the two free valencies of the carbon atom, bring
it again close to the carbon. At this moment the oxygen becomes
again doubly linked and moves off into greater distance, and the
former play commences anew. The condition, therefore, which
the above mentioned critic believes to be missing, is actually
present, namely: when the hydrogen atom has moved back,
passed the mean position, and approximated itself to the oxygen
atom, the opposite phase becomes superiorily attractive.” Both,
the hydrogen atom and the oxygen atom are continuously in a
fierce state of vibration, as may be indicated by the following
formule :

ZO 2O O
es x | la / |
= —C Vy = Cees
\ Ss
H H
(a) (b) (a) (b)

The lability, ze. the kinetic chemical energy in an alde-
hyde group, under certain conditions, may reach a much higher
intensity. Such a condition is, ¢.g., the presence of an amido-
group in the same molecule. The continuously moving oxygen

(1) Oxygen, in the form of hydroxyl occupies a volume of 2.3; while in the car-
bonyl form one of 5.5. Also doubly linked carbon atoms havea larger molecular volume
(and higher thermic value) than simply linked ones.

(2) Attention may here be called to the inference drawn by Zhomson (Ber. 19, R.
76) from thermo-chemical observations, that aldehydes behave in certain points like com-
pounds containing a hydroxyl-group besides unsaturated carbon ; this would correspond
to the phase (b) of our inference drawn from a very different contemplation.

LABILITY AND ENERGY IN RELATION TO PROTOPLASM. 403

atom of the aldehyde group sets the hydrogen atoms of the amz-
do-group into rapid motion, as,—to give a rough analogy
moving magnet held beneath a sheet of paper moves iron
particles spread upon the sheet, which cannot unite with the
magnet, the sheet holding them back. These moving hydrogen
atoms of the amido-group, again, will exert an influence upon the
labile hydrogen atom in the aldehyde group which, in con-
sequence of the repulsion it suffers, is compelled to increase
the amplitude of its oscillations.” Thus we can understand why
the lability of an amido-aldehyde is very great and the inclination
to spontaneous changes much more marked than with common
aldehydes. The degree of lability, however, is much influenced
by the more or less saturated condition of the compound, the re-
lative position of the labile atoms, and the relative number of the
CH,-groups. Thus, amido-ethylaldehyde is much more labile
than the 6-amido-valeraldehyde ; the latter remaining even un-

a

changed on distillation under diminished pressure, while the
former changes soon after it is liberated from its combination
with acids. The amido-benzaldehydes, again, are less labile,
although reacting with great readiness and readily changing
when brought in contact with dilute hydrochloric acid.

Such a state of kinetic chemical energy was foreseen by.
me to exist in the active albumin, which I consider a pro-
duct of the condensation of the di-aldehyde of aspartic acid (cf.
Bulletin II, No. 2). The active group a which, by atomic mig-
ration, easily passes into the passive group f, would, according
to my deduction, have the following structure :

—CH—NH, —CH—NH
| | Us,
=C—CHO =C—CHOH
—_—_—_—— ———
a p
Aldo-amido-structure. Hydroxyl-imido-structure.

The one essentia! to my theory is the constitution of the labile
(active) group and the conclusion that the active albumin is a
product of the condensation of aspartic aldehyde, which may be
built up in the living plant cells of formaldehyde and ammonia; a

(1) Hydrogen atoms of a high degree of lability behave sometimes somewhat like
nascent hydrogen, leading to powerful reductions.

ry

404. LABILITY AND ENERGY IN RELATION TO PROTOPLASM.

non essential is the supposed relative number of the active
groups (twelve for Lieberkuehn’s formula), as unforeseen atomic
migrations may occur. Compounds with more than two aldehyde
groups” have not thus far been prepared but this fact in itself
gives no ground to infer that molecules with more than two
aldehyde groups are impossible. The actual preparation of a
compound consisting of six keto-groups, vzz., the triquinoyl,
appears in itself hardly more striking than, e.g., that of a ben-
zene-ring would be to which six aldehyde groups are attached, cor-
responding to the as yet hypothetical aldehyde of mellitic acid.

When, in the year 1880, I explained before the Munich
Chemical Society my theory of the formation of albumin in plants,
and [had expressed the opinion that amido-aldehydes, an unknown
class of bodies then, would certainly be prepared at a future time,
some of the chemists present denied the possible existence
of such bodies, as they would change at the moment of prepa-
ration. Later, however, amido-aldehydes were prepared, and it
was found that the degree of lability varies considerably with
different amido-aldehydes. Moreover, the toxicological facts
described (Bulletin II. No. 1), give colour to my deduc-
tion that the lability of the living protoplasm is caused by the
co-existence of aldehyde and amido-groups in the active albumin.

Lo recapitulate: Labile (kineto-labile) compounds, which
readily change into an isomeric stable modification contain certain
atoms loosely bound, which condition is caused by a certain
amount of kinetic chemical energy counteracting the force of affi-
nity existing between the atoms of the compound. Every energy
of the kinetic kind is motion, and motion can be conveyed from
one body to another. Kinetic chemical energy (atomic motion)
may, by being conveyed to atoms of other easily changeable
compounds, lead to chemical changes in them (katalytic pheno-
mena). I have attempted to give an explanation of the contz-
nutty of such motions in aldehyde groups, according to our pre-
sent state of knowledge. But even those who may consider
that explanation still incomplete will at least admit the cor-
rectness of the deduction that labile atoms are in a special state of
continuous vibration.

For his convenience, the reader may now follow a juxta-
position of inferences of my theory and of actual observations:

{(1) Cf. On phthalic aldehyde, Ber. Chem. Ges. 20, 509.

LABITITY AND ENERGY IN RELATION TO PROTOPLASM.

Theory.
We
Albumin is formed by con-

densation of the still hypothe-
tical aspartic aldehyde which
in plant cells either is produced
from asparagine or built up of
form-aldehyde and ammonia.

2re
There is a chemical differ-

between the albmmnin of
the living and that of the dead
protoplasm.

ence

3.
The labile, active
leads by organization to living
matter, as such and inthe form

of nuclein and nucleo-albumin.

albumin

4.
The lability of the albumin

of the living protoplasm is
caused by the presence of alde-
hyde and amido-groups.

5.

The conversion of the albu-
min of the living to that of the
dead protoplasm, presents a
remarkable analogy to the
change of a labile substance
into a stable modification.

405

Facts.
Ne
There exist intimate physi-
ological relations between as-
and the
former is an excellent material

paragine albumin ;

for building upthe latter. The

formation of albumin often

takes place with great rapidity.
BD

The living protoplasm shows
a chemical behaviour totally
different from that of the dead.
be
There frequently occurs, as
reserve-material, in
highly labile kind of albumin
of aldehyde character, whose

plants a

chemical nature is altered by

the same influences, as those by

which the protoplasm is killed.
4.

Compounds which react upon
aldehydes, and such as react
upon labile amido-groups with
great energy, are poisons for
all organisms.

5.

The transition of living pro-
toplasm into dead is accompa-
nied by contraction and deve-
lopment of heat.

The unbiassed reader will perhaps find here some coinci-

dence between theory and facts.

(1) A full account of theoretical views and actual observations is contained in my

treatise :

Tritoner & Co:

The Energy of Living Protoptasm, London, 1896; Kegan Paul, Trench,

On the Formation of Mannan in Amorpho-
phallus Konjak.
BY
Michito Tsukamoto, Nogakushi.
Professor of Chemistry in Harris’ Science School, Doshisha, Kyoto.

In regard to the formation of carbohydrates, A morphophallus
koigak is of special chemical interest. The tuber of this araceous
plant, from which different articles of food are prepared in Japan,
contains, as Z’szyz has shown, no starch, but a very large pro-
portion of mannan. Afterwards K7znoshita found that the
tuber contains two mannans, a slimy soluble one and another
insoluble in water. The question suggested itself to my mind : is
this mannan produced from glucose by transformation in the
tuber, or is the soluble mannan already formed in the aves from
glucose, or finally is mannose a direct product of the assimilation
of carbonic acid in the place of glucose in this plant? Thus far,
mannose has never been found in plants except in the form of
anhydrides, and it would be no doubt of great physiological in-
terest if it could be shown that there exist chlorophyll bodies
which produce mannose instead of glucose.

I examined, therefore, the leaf of this plant and found but
very little starch,” besides globules not coloured by iodine, and
raphides. Further I observed that all portions of the leaf
contain a very slimy substance which proved to be an anhydride
of mannose. The extract prepared with warm water (40°-50° C)
was mixed with a large proportion of alcohol and the resulting
flocculent precipitate was purified by again dissolving it in water
and precipitating once more with alcohol. The aqueous solution

(1) This Bulletin, vol. II., No. 2. (1894).

(2) This Bulletin, vol. II., No. 4. (1895).

(3) I have examined small fragments of the leaf-blade, coilected one clear sunny
afternoon, with iodine solution under the microscope and found but very few granules
were coloured blue. In the ribs there was also only a small amount of starch but
relatively more than in the blade-cells. The cellulose walls in the blade gave the
ordinary cellulose reaction with sulphuric acid and iodine.

(4) The leaf stalk and larger portions of the ribs were separated from the leaf blade,
including smaller ribs, and both were separately investigated.

FORMATION OF MANNAN IN AMORPHOPHALLUS KONJAK. 407

of this precipitate had a slimy consistency, and showed the in-
teresting property of Josing its slimy character on prolonged
boiling, whereby the slimy compound separated as zuxsoluble floc-
cult, The freshly prepared solution of the mucilage yields a
white flocculent precipitate with basic lead acetate upon the ad-
dition of some ammonia, and a thick blue precipitate with either
Fehling’s solution or copper sulphate solution in presence of
sodium hydrate. On boiling with dilute sulphuric acid of 3 °/,
the’mucilage was, after a few hours, transformed into a sugar,
which, after the removal of the sulphuric acid by barium carbon-
ate and evaporation of the filtrate, at once yielded, on the addition
of phenyl-hydrazine acetate, the characteristic precipitate of
mannose-phenylhydrazone.” To test whether galactans or
pentosans were present in stalk and blade, the residue, which
remained after the extraction of several hundred grams with
alcohol and warm water, was boiled for several hours with dilute
sulphuric acid of 4°/,. The syrup obtained after neutralizing
with barium carbonate and evaporating, was tested for the
presence of pentose with phloroglucin and hydrochloric acid, and
for galactose by evaporation with nitric acid, but neither a pentose
reaction nor mucic acid were obtained ; therefore wetther pentos-
ans nor galactans were present. Only a little mannose was
obtained from this insoluble part.

The question whether mannose as such is present in the
stalk and blade I tried to answer by extracting these objects
with alcohol of 50°/, whereby the mannans would remain in-
soluble, while the sugar would be dissolved. These extracts were
evaporated,” dissolved in a little water, and basic lead acetate
added to remove tannin and other impurities. The filtrate freed
from lead with hydrogen sulphide was evaporated after neutrali-
zation with sodium carbonate. Upon the addition of phenylhydra-
zine acetate, only the extract of the stalk yielded a sufficient quan-
tity of the precipitate of mannose-phenylhydrazone, while there
was a doubtful trace in the case of the extract of the blade. The
filtrate of the above precipitate yielded, upon further addition of
phenylhydrazine acetate and heating on the water bath, sucha

(1) This mucilage of the stalk and blade agrees therefore in its essential properties
with the soluble mannan which Azxoshita obtained from the tuber (loc. cit.),

(2) Since in the case of the stalk an acid reaction of the extract was noticed, it was
neutralised with sodium carbonate.

408 PORMATION OF MANNAN IN AMORPHOPHALLUS KONJAK.

considerable quantity of phenylglucosazone that I must conclude,
there was present in the stalk, besides mannose, glucose or
fructose,” or both to some extent.®) The mannose-phenylhy-
drazone first obtained was recrystallized and showed the proper
melting point of 195°C and the characteristic tabular crystalline
forms. It was, like pure mannose-phenylhydrazone, hardly
soluble in absolute alcohol and warm acetone, almost insoluble
in ether and benzene, and reduced Fehk/ing’s solution very
strongly upon warming.

The fact that the slimy mannan is present in the cells of the
leaf makes it highly probable that it plays to some extent the
role of starch in this plant, but whether mannose is really the
first product of assimilation, can not yet be answered. I hope
however to settle this question by further investigations. On the
other hand, the presence of mannose as such in the stalk ts evia-
ently of high physiological interest as this ts the first time that tt
has been found as such in plants. 1 am indebted to my most
honoured former teacher, Dr. OSCAR LOEW, Professor in the
Imperial University, for kind suggestions.

(1) A portion of the above syrup obtained from the extract of the stalk was tested for
fructose with resorcin and concentrated hydrochloric acid, whereby a cherry red colou-
ration resulted. Another portion of the syrup was treated with a mixture of ether and
alcohol ; on evaporation of the extract I obtained the proper red colouration with the
above reagent. It is therefore, probable that some fructose was present in the stalk.

(2) C. A. Lobry de Bruyn (Rec. Trav. Chim. 14 (1895) 201-206 and Ber. Chem.
Ges. 28 (1895) 3078-3082) has recently discovered the highly interesting fact, that by
simply treating with alkalies these three sugars can be transformed into one another.

On the Formation of Asparagine in Plants under
Different Conditions.

BY

U. Suzuki, Nogakushi.

Although the question of asparagine production in plants
has often been the object of investigation, many points are still
left unsettled, and in certain respects, even contradictory state-
ments, not only in regard to its production, but also in regard to
its transformation into proteids, are met with. The production
of asparagine from proteids during the germination process has
been quantitatively followed up by E. SCHULZE, but thus far, all
the explanations attempted have proved unsatisfactory ; therefore
Dr. O. LOEW has proposed a new view which seems to accord
better with various facts.” According to this view asparagine is a
synthetical product, into which the ammonia of the decomposed
proteids is transformed. Recently some experiments have been
made by Mr. KINOSHITA in the laboratory of the Agricultural
College, Imperial University, which show that the ammonia
taken up by the roots may also pass into asparagine ; but as these
experiments were made with only two kinds of Graminee it
appeared to me necessary, in view of the importance of the ques-
tion, to carry out a larger number of experiments, to test whether
this transformation takes place in different families and in dif-
ferent states of development. Furthermore, I desired to compare
quantitatively the asparagine production from different ammonium
salts” and from nitrates, and to determine how the results are
affected by the increase of sugar. These experiments are des-
cribed in the following pages, while the analytical data follow in
an appendix.

I. Experiments with sun-flower (Helianthus annuus ).

The young sun-flower plants, 30-40°™ high, were taken very
carefully from the field, and after washing the roots, placed in
the following solutions kept in a glass house :

(1) Cf. The Energy of Living Protoplasm, London, 1896. p. 38.
(2) In some cases I also applied urea.

410 SUZUKI ;

a, 0.1 % solution of ammonium nitrate.
b, i " ,, ammonium chloride.
Ci O!2ae be ,, sodium nitrate.
d, distilled water.
Time :—S8 days. (Oct. 27—Nov. 4.)
Temperature :—Min. 10.5°C.; Max. 40°C.
Neither the solutions of sodium nitrate, nor the plants grown
in the different solutions, gave any reaction for ammonia ; only a
few leaves withered during the experiments, and these were re-
moved. The result of the analysis was as follows.
Table I :—In 100 parts of dry matter :

Plants in
Original() plants Control(?) plants Ammonium Ammonium Sodium
(Oct. 27th) (Nov. 4th) nitrate chloride nitrate
Asparagine nitrogen 0.14 0.29 0.78 0.99 0.39
Asparagine(?) 0.64 1.38 3.67 4.67 1.85

This result shows that ammonium chloride produced much
more asparagine than ammonium nitrate, and this again double as
much as sodium nitrate, while an increase of asparagine in the
control case, compared with the original plants, taken from the
field, must evidently be due to the gradual transformation of
nitrates that had been stored up in the stem and roots, whose
presence had been shown by the diphenyl-amine test.

II. Experiments with yellow-lupine (Lupinus luteus).
Young plants 20°" high were taken from the field ; 4-6
plants were placed in 300° of the following solutions :—

a, 0.1% solution of urea.
; . 3 ,, ammonium phosphate.
CF 102% a ,, sodium nitrate.
d, distilled water.
and kept in the glass house for 6 days, from Nov. 7th to 13th.
Temperature :—Min. 8°C. ; Max. 35°C.
After drying, the entire plants were used for analysis.
Table II :—In 1009 parts of dry matter.

(1) “Original” means the plants which were analyzed at the beginning of the
experiments,

(2) “Control”? means the plants kept in distilled water.

(3) The water of crystallization of asparagine is not included in these calculations.

FORMATION OF ASPARAGINE IN PLANTS. 4II

Plants in

ald = P IEEE A eS
Original plants Control plants Ammonium Urea‘!) Sodium

(Nov. 7) (Nov. 13) phosphate nitrate
Asparagine nitrogen 0.38 0.40 0.62 1.18 0.72
Asparagine 1.76 1.91 2.90 5-55 3-35

It is to be remarked in this case that urea, which might have
been transformed into ammonium carbonate in the plants, was
much more favourable than sodium nitrate for asparagine pro-
duction, while the ammonium phosphate was less favourable than
sodium nitrate.

Ill. Experiments with ‘“‘ sendan” (Alelia Faponica).

The young branches of this plant were put into flasks con-
taining about 250° of the following solutions :—

a, 0.1 % solution of ammonium chloride.

|5), ea C ,, ammonium phosphate.
CaO 0% af ,, sodium nitrate.
d, distilled water

and kept in the glass house for 6 days (Nov. 7—Nov. 13).’?
Temperature :—Min. 8°C. ; Max. 35°C.
After drying, the leaves and 20°" of the upper parts of the
stems were analyzed.
Table III :—In 100 parts of dry matter :

Plants in

a
Original plants Control plants Ammonium Ammonium Sodium

chloride phosphate nitrate
Asparagine nitrogen 0.11 0.13 0.37 0.29 027
Asparagine 0.52 0.59 1.76 1.38 1.25

IV. Experiments with squash (Cucurbita melo peppo).

Squash seeds were distributed in three large pots containing
sea sand washed first with hydrochloric acid, then with common
soft water ; the pots were kept in the glass house, in which the
temperature ranged from 22°C. to 44°C. Four days after sowing,
the germination had fairly set in, and when 3°™ high, the shoots of
one pot were treated with 0.05 % solution of ammonium nitrate,

(1) The nourishing solutions did not show any development of bacteria ; the plants
remained healthy. About 4/, of the total urea of the solution was here transformed
into asparagine.

(2) At the end of the experiments the plants began to suffer, the temperature having
been too high.

412 SUZUKI ;

of the second with 0.1% solution of sodium nitrate, while those
of the third served asa control case. Ten days after the treatment,
the plants were removed from the seed bed,” dried, and only
the stems and roots used for analysis, because it has been found by
others that cotyledons and leaves always store up less asparagine
than stems and roots.

Average height of the control plants® 133
a ,, plants in ammonium nitrate 10.1°™"
ie ,, plants in sodium nitrate 6.2c
Average fresh weight of control plants 2.36 grams.
2 =, * of the plant in ammonium
nitrate 1.79 grams.
ma i ¥ of the plant in sodium
nitrate 1.20 grams.

The total ammonium nitrate solution applied was 318°°=
0.0557 gram nitrogen.

The total sodium nitrate solution applied was 322°°=
0.053 gram nitrogen.

The aqueous extract of a portion of these plants did not show
a trace of ammonia, while the reaction for nitrates was obtained
in the plants in ammonium nitrate and sodium nitrate with Kxop’s
solution.

Table IV :—In 100 parts of dry matter free from ash.

Plants in
Control plants Ammonium nitrate Sodium nitrate
Albuminoid nitrogen 2.49 2.58 2.53
Proteids 15.59 16.13 15.83
Asparagine nitrogen 1,38 3.94 3933
Asparagine 6.53 18.57 15.69(3)

The same experiment was repeated later in the autumn with
shoots a little more developed than in the first case.

(1) Some of the plants had been attacked by afungus, Fusarium Lateritiumt, especi-
ally in the lower parts of the stems, and in the cotyledons; such plants were, of course,
discarded.

(2) The better growth of the control plants in this case is evidently due to the very
early stage of the shoots, which contained a large quantity of soluble nutrients from the
cotyledons, Thus an increase of soluble nutrients had a retarding effect. Ina 10 °/)
cane sugar solution, e.g , young plants will not develope so well as in one of I 9/, only.

(3) The cause of the considerable production of asparagine from nitrates may in
this case be due to the high temperature, favouring the reduction of the nitrates by the
increased amount of sugar present.

FORMATION OF ASPARAGINE IN PLANTS. 413

Time of experiments (duration). Sept. 19th—Oct. 8th.
Beginning of germination. Sept. 24th.
Solution applied. Oct. 2nd—Oct. 8th.

Temperature in the glass house :

Min 17o@ar Wax 44°@:

Average height of control plants. Siar
he if ,, the plantinammonium nitrate. 10.11%
- » 5, the plant in sodium nitrate. O35 0:
Average fresh weight of a control plant. 1.07 gram.
Bs ue st ,, the plant in ammonium
nitrate. 1.13 gram.
i “ si ,, the plant in sodium
nitrate. 1.18 gram.
Ammonium nitrate solution applied. 508°" =0.178 gram.
Sodium “, - 4a 608° =0.201 gram.

Table V :—In 100 parts of dry matter (free from ash).

Pants treated with ;

Control plants Ammonium nitrate Sodium nitrate

Albuminoid nitrogen 2,82 2.99 Brin
Proteids 17.64 18.66 19.44
Asparagine nitrogen 1.65 2.57 2.33
Asparagine 7-77 12.10 10.98

V. Experiments with potato plants (Solanum tuberosum).

Well developed potato plants were taken from the field, and
placed in the following solutions :—

a, 0.1% solution of ammonium nitrate.
b, 0.29 solution of sodium nitrate.
c, distilled water.

The plants were kept in a glass house.

Time of experiments. Oct. 12th—Oct. 21th.

Temperature:— Min. 16°C.; Max. 40°C.

At the end of the experiment, the plants began to suffer,
and a portion of the roots commenced to decay. Of course, all
the unhealthy parts were removed ; only the lower portion of the
stem 10-15°™ in length served for analysis. No ammonia was
found either in the original plants or in the other three cases, but

(2) In the second experiments with squash, the plants were also atiacked by a
fungus; hence I could not continue the experiment.

414 SUZUKI ;

a moderate quantity of nitrates was found in all cases. <A few
drops of the juice of the plants in ammonium nitrate were left to
dry up on an object carrier, and yielded besides some long
needles (probably potassium nitrate), numerous asparagine cry-
stals readily recognized under the microscope by their insolu-
bility in cold saturated asparagine solution.

The result of the analysis was as follows :—

Table VI. In 100 parts of dry matter.

Plants in
SS
Original plants Control plants Ammonium Sodium
(Oct. 12) (Oct. 21) nitrate nitrate
(stem) (roots) (stem) (stem) (stem)
ns
Asparagine nitrogen 0,20 0.22 0.75 1.51 0.98
Asparagine 0.92 1,02 3-54 7.06 4.61

We observe here that ammonium nitrate was much more
favourable for asparagine production than sodium nitrate, and
that the control plants kept in water contained much more aspar-
agine than the original plants taken from the field, which fact may
be due to the transformation of nitrates, already present in the
plants, into asparagine.

Another experiment with potato plants was made on an open
farm. The plants 30-40°™ high were irrigated with :—

a, 0.1% solution of ammonium phosphate.
by, 0.2% 33 ,, sodium nitrate.

There was no rain-fall during the experiment, but sometimes
the temperature dropped down to 3°C. at night. After 6 days

(Nov. 7th—Nov. 13th), the plants were removed and the entire
plants were used for analysis.

Total ammonium phosphate solution applied. 300°°
,, sodium nitrate sd 300°°°

Table VII. In 100 parts of dry matter.
Plants treated with

ee

Original plants Ammonium phosphate Sodium nitrate
Asparagine nitrogen 0.57 0.37 0.53
Asparagine 2.68 1.76 2.53

In this case we observe no increase of asparagine after the
treatment with sodium nitrate, and even a decrease by the treat-
ment with ammonium phosphate. The conditions for the forma-
tion of protezds from asparagine were evidently very favourable
and especially promoted by ammonium phosphate.

FORMATION OF ASPARAGINE IN PLANTS. 415

VI. Experiments with potato shoots.

A. On April 2nd, etiolated potato shoots which had grown
in moist saw dust in the dark, were placed in the following solu-
tions :—

@, 02% urea solution.

6, 0.2% sodium nitrate solution.

c, 0.2% urea and 2% sugar solution.

d, 0.2% sodium nitrate and 2% sugar solution.

é, distilled water.

The plants were kept in a dark room for 6 days (April 2nd
to April 8th).

The shoots were 12-20° long, and weighed 2 grams on an
average in the fresh state.

Although they had not grown for 6 days they were sfill quite
healthy ; the solutions had remained almost entirely clear.
Table VIII :—In 100 parts of dry matter :—

Plants in
ae oe

Original Control Te Sodium Urea and Sodium nitrate

plants. plants. nitrate. sugar. and sagar.
Albuminoid nitrogen 2.18 1.90 I.gI 1.79 2.11 1.80
Asparagine nitrogen 0,68 0.98 1.65 0.97 1.38 1.18
Asparagine Py 4.62 7.78 4.58 6.51 5-50

B. On the 8th of April, the same experiment was repeated but
this time, in full day light.

Length of shoots, 12-24°"

Average fresh weight, 1.848""™

Time of experiments, April 8th—April rsth.“?
Table I1X.:—In 100 parts of dry matter :—

Plants in

va

Original Control Sugar- Urea Sodium — Sodinm nitrate

plants. plants. nitrate. and sugar.
Total nitrogen 3.14 3.29 2.74 ae 3.48 3.04
Albuminoid nitrogen 1.80 1.86 1.45 1.97 1.89 1.80
Asparagine nitrogen 0,55 0.70 0.75 1.70 0.78 0.56
Nitrogen in nitrates fo) oO fe) fo) 0.21 0.16

As the application of sugar causes a remarkable increase
of dry matter in the plants, the percentage of nitrogen is lowered
considerably. Therefore, for the sake of correct comparison, I
calculated the following table :—

(1) The lower parts of a few shoots were removed, having suffered somewhat.

416 SUZUKI ;

Table X:—In 100 parts of total nitrogen :—

Plants in _
———
Cine, pak See) Ure eee
Total nitrogen 100 bfere) 100 100 100 100
Albuminoid nitrogen 57.3 56.5 ze 53-3 54.6 59.2
Asparagine nitrogen 17.5 213 27.4 45.5 22.4 184
Nitrogen in nitrates 0 o fe) fe) 6.0 5.2

We observe here no considerable decrease of albuminoid ni-
trogen during the experiments, but a little increase of asparagine
nitrogen takes place in the control and sugar case; but as this
increase does not correspond with the decrease of albuminoid
nitrogen, it must be due to the transformation of other amido-
compounds into asparagine. We observe here alsoa very great
difference of asparagine nitrogen in the urea and sodium nitrate
plants. Sodium nitrate does not here seem well suited to be-
come asparagine. Sugar had no great influence upon the
increase of albuminoid nitrogen; this must be due to the want
of some otlier necessary conditions for the protein formation
(want of sulphates ?).

Isolation of asparagine crystals: 3.27°°"* air dry plants
treated with urea yielded 0.15%" by crystallization, while the
calculation requires about 0.27°%". In this case, I could not
separate the slimy mother liquor which prevented the cry-
stallization of all the asparagine.

VII. Experiments with “ £eyasagara” (Holesia hispidum).

Young branches covered with leaves, 25-30°" in length,
were placed in the following solutions :—

a, 0.1% solution of urea.

b, 3 5 wy.» 2nd 10% cane sugar

c, 0.1% solution of ammonium phosphate,

a, = ne = if and 10% cane
sugar.

e, distilled water.

The objects were kept in the dark, for 7 days (Oct. 31st—
Nov. 7th). No ammonia was found stored up after the treat-
ment and the leaves remained generally very healthy.

After drying, the leaves only were subjected to analysis :-—

(1) As the total nitrogen in some of the above cases was increased absolutely, the
albuminoid nitrogen was apparently decreased.

FORMATION OF ASPARAGINE IN PLANTS. 417

Table XI :—In 100 parts of dry matter :—

Plants in

4

= <
Original Control Amm. Amm. phosphate ie: Urea and
plants. plants. phosph. and sugar. ¥ sugar.

Asparagine nitrogen O.11 0.15 0.16 0.10 0.23 0.12

Asparagine 0.51 0.72 0.73 0.48 1.10 0.55

In this case it is very evident that the introduction of sugar
prevented the increase of asparagine.”

VIII. Experiments with buckwheat plants (Polygonum
Jagopyrum).

Experiments were made with buckwheat plants taken from
the farm in full blossom and dcaring seeds. There was only a
trace of asparagine and a moderate amount of nitrate present,
but no trace ofammonia. Preliminary experiments showed further
that in this case, some of the ammonia offered as ammonium
nitrate in 0.05% solution, was found afterwards as such in the
stems to the extent of 0.1796” of the air dry matter.

The plants were now placed in the following solutions :—

a, 0.1% solution of ammonium nitrate.

6, 0.1% solution of ammonium chloride,

¢, 0.2% solution of sodium nitrate.

dad, distilled water.

The plants were kept in the glass house for 9 days (Oct.
12th—2Ist.).

Temperature :—Min. 16°c ; Max. 40°%c.

All the plants were healthy and the solutions remained
clear.

The analysis for which only stems and leaves were used
yielded the following results :—
Table XII. In 100 parts of the fresh plants :—

Plants treated with

7
Control Ammonium Ammonium Sodium.
plants. chloride. nitrate. nitrate.

Asparagine nitrogen and

nitrogen in ammonia. 0.05 0.13 0.08 0.06

Nitrogen in ammonia. fo) 0.08 0.04 0)

Asparagine nitrogen. 0.05 005 0,04 0 06

Asparagine 0.25 0.22 0.18 0.29

(1) A small amount of sulphates was found in the leaves which might have been
utilized in the production of new proteids from asparagine and sugar.
(2) Cf. Analytical data in the appendix.

418 SUZUKI ;

The reason why so little asparagine was formed here and
the ammonia taken up remained as such partly preserved in the
plant, may be due to the fact that the development of young
seeds required most of the sugar produced by the leaves, for
the production of starch, and therefore the necessary amount
of sugar for the production of asparagine was not available. In
the following experiments, therefore, all the flowers and seeds
were vemoved before the treatment commenced ; the plants
were placed in the following solutions ;—

a, 0.1% solution of ammonium nitrate.

b, = a va - and 10% sugrr.
C, 3 x5 ty chloride and 10% sugar.
a, * a es carbonate.

e, D ss ia rf and 10% sugar.
Ws 55 i - phosphate.

Ee x 7 ie ts and 10% sugar.

These were kept in the glass house for 11 days (Oct. 24th—
Nov. 4th).

Temperature :—Min. 10.5°c.; Max. 37°c.

In this case, the result differed considerably, no trace of am-
monia being found in any of them.

The analysis for which only the stems and roots were used,
yielded the following results.

Table XIII :—

Plants treated with
ee ——— SS

mm.
Ammo- Ammo- Amm, Ammo- Amm Amm.
Ammo- —. : = - phos- °4
. » Niumni- nium carbon- nium chloride
nium ni- z phate
trate and carbon- ate and phos- and
trate. and
sugar. ate. sugar. phate. J sugar.
‘ : sugar.
Asparagine nitrogen, 0.39 0.10 0.48 0.27 0.26 1.10 0.10
Asparagine. 1.82 0.48 2.25 1.27 1.20 0.46 0.50

We sce from these results, that the formation of asparagine
from ammonia was considerable. ‘The sugar produced from the
leaves evidently yielded the carbon for the asparagine but
we see also on the other hand, that, in those cases where sugar
was added to the solutions, the amount of asparagine was again
less, which finds a natural explanation in the fact that, by a larger
excess of sugar, the protein production was so stimulated that

FORMATION OF ASPARAGINE IN PLANTS. 419

the asparagine was again for the greater part consumed.” Sugar
inthe nourishing solution therefore not only prevents asparagine
production from proteids, as Monteverde has ascertained with
Syringa but also lowers the amount of asparagine produced
from ammonium salts offered to the roots !

IX. (4.) Experiments with “d@zkawashima-na”
(Brassica campestris).

This experiment was made on the field, the plants about
30° high with as yet very small roots were irrigated with the fol-
lowing solutions :—

a, 0.1% solution of ammonium phosphate.

ie TOY 7, ,, sodium nitrate.

Duration of experiments, 8 days (Nov. 20th—Nov. 28th).
Total solutions applied for each case, 600 c.c.
The entire plants were used for analysis.
Table XIV. In 100 parts of dry matter :—

Plants in

7x

— -—_—_—_—.
Control Ammonium Sodium
plants. phosphate. nitrate.

Asparagine nitrogen 0.85 0.55 0.83

Asparagine 3-99 2.60 3.90

(5.) Experimets with Brassica Campestris, var,
‘* Swedish turnip.”
Full grown plants with large round roots, were removed
from the farm, and placed in the following solutions :—

a, 0.1% solution of ammonium nitrate.

Gre O02 im 15; ,, ammonium carbonate.
GE OL2% me ,, sodium nitrate.

d, distilled water.

The plants were kept in the glass house.
Time of experiments :—7 days (Oct. 16th—Oct. 23rd).
Minwivjcer. Max. 40°.

Temperature :

At the end of the experiments, the plants had suffered very
much, the temperature being too high, and a portion of the roots
died. After washing the roots and removing the dried lcaves,
only the healthy parts were analyzed which yielded the following
results :—

(1) The presence of sulphates, which I found in the leaves and stems, no doubt,
rendered this process possible.

420 SUZUKI ;

Table XV. In 100 parts of dry matter :—

Plants treated with

ee
Original Control Ammonium Ammonium Sodium
plants. plants. nitrate. carbonate, _ nitrate.
Asparagine nitrogen 0.65 1.51 1.69 1.69 2.04
Asparagine 3-07 7.11 7.99 7.97 9.62

Here, in the control plants, very much asparagine was
formed, this can only be due to the transformation of nitrates,
originally stored up in the plants, aided by the presence of much
sugar. Indeed in a second experiment the decrease of nitrates
stored up was plainly observed during this asparagine produc-
tion; and in this case even less asparagine was noticed in the
plants nourished with ammonium salts. In these experiments,
five plants of similar size,” were placed in the following solu-
HONS ==

a, 0.19% solution of ammonium chloride (1.5 litre).

OM as »» 3 ammonium phosphate (1.5 litre).
Ee 10.2% a ,, sodium nitrate C - oeee
d, distilled water. C Se ate

They were kept in the glass house.
Time of experiments :—7 days (Nov. 6-13).
Temperature :—Min. 7°c.; Max. 36°.
After drying” only the healthy stems and leaves were ana-
lyzed, with the following results.
Table XVI. In 100 parts of dry matter:
Plants treated with

er

Original Control Ammonium Ammonium Sodium

plants. plants. chloride. phosphate. nitrate.
Asparagine nitrogen 0.55 1.47 1.25 1.15 I.II
Asparagine 2.57 6.92 5.90 5-40 5 21
Nitrogen in nitrates. 0,83 0.41 -- — —

(c). Experiments with Brassica campestris,
var. ““Zamna’™©
Three experiments were made with this variety.
I. Experiment (made on the field).

The young plants growing on the farm about 20™ high were
irrigated with the following solutions :—

(1) The roots weighed 200 grams on an average; the leaves were 30-50 cm. long.

(2) At the end of the experiments the plants i sodium nitrate and ammonium
phosphate began to suffer. The other plants were quite healthy.

(3) This variety has very soft juicy leaves and very small roots.

FORMATION OF ASPARAGINE IN PLANTS. 421

a, 0.2 % solution of ammonium phosphate.
6, 0.4% e ,, sodium nitrate.

Time of experiments, Nov. 23rd-28th.
Total solution applied :

Ammonium phosphate 200 cc.
Sodium nitrate 200 CC.

After drying, the entire plants were analyzed.
Table XVII. In 100 parts of dry matter :-—

Plants treated with

—————————_———————_——————
Control plants Ammonium phosphate Sodium nitrate

Asparagine nitrogen 0.39 0.47 0.42

Asparagine 1.85 2.21 2,00

i Experiment.

The plants were carefully removed from the field and placed
in glass cylinders, each containing 500°" of the following solu-
tions, and kept in the glass house :

a, 0.2% solution of ammonium chloride.
3 ,, sodium nitrate.

C, distilled water.
Duration of experiment: 7 days (Nov. 30th—Dec, 7th).
Temperature :— Min 32> Max. 33°C.

Total nitrogen absorbed by ;—

a, the plants in ammonium chloride was 0.0462 grams=
0.5% of dry matter.”

b, the plants in sodium nitrate was 0.0437 grams=0.59%
of dry matter.”

The plants remained normal and the solutions quite clear.

The analysis, for which the entire plants were used, yielded the
following results :—

Table XVIII. In 100 parts of dry matter :—

Plants treated with

——[—[{S=_——
Original Control Ammonium Sodium
plants plants chloride nitrate

Total nitrogen 4.60 444 4.83 4.78

Albuminoid nitrogen 1.65 1.63 1.86 1.35

Asparagine nitrogen() 0.28 0.71 0.86 0.95

Nitrogen in nitrates 1.04 0.36 0.62 0.88

(1) Total dry matter of the plants in ammonium chloride (at the end of the ex-
periments) was 9.328 grams.

(2) Total dry matter of the plants in sodium nitrate (at the end of the experiments)
was 7.47 grams,

(3) As I had suspected the presence of some organic bases like arginine, etc , I em-
ployed first phospho-tungstic agid to remove them. Indeed, the result for asparagine was
then considerably lower than by the usual method, while in most of the other cases, I
found no essential difference by the previous application of phospho-tungstic acid.

422 SUZUKI ;

Here also it is seen that the increase of asparagine nitrogen
is accompanied with the decrease of nitrates in the plants, and
that in this case the asparagine is not a decomposition product of
proteids.

The relation of these numbers will become still clearer, if
we add the following table :—

Table XIX. In 100 parts of total nitrogen.
Plants treated with

AES a

Original Control Ammonium Sodium

plants plants chloride nitrate
Total nitrogen 100 100 100(1) 100/1)
Albuminoid nitrogen 35-9 36.7 42.9 322
Asparagine nitrogen 69 16.0 20.0 22.5
Nitrogen in nitrates Py | 8.1 14.3 21.0

III. Ixperiment.

Here, the influence of sugar and that of temperature upon
the result was to be observed. The plants were taken from the
same farm as before, and placed in glass cylinders, containing
each 500° of the following solutions :—

a. 10% solution of sugar.

bz 402s ,, ammonium chloride.
CG 55 Sp 2 55 and 10% sugar.
d. Bi sat ,, sodium nitrate.

e re as rr i re and 10% sugar.
f. distilled water.
e 10% sugar solution.
h. distilled water.
a, b, c, d, e, f, were kept in the glass house, while g, and h,
were kept in the laboratory, where the temperature was much
lower than in the glass house.”

Duration of experiment, 8 days (Dec. 10th—18th).
Temperature in the glass house :—Min. 1°C ; Max. 38°C.

a ,, 5, laboratory :—Min. 5°C ; Max. 15°C.
Plants treated Ammonium Ammonium chloride Sodium Sodium nitrate
with chloride and sugar nitrate and sugar
Total nitrogen absorbed Z i
inverame 0.042 0.038 0.041 0.037
Percentage of absorbed
nitrogen in the dry 0.53% 0.31% 0.54% 0.34.%
matter($)

(1) The nitrogen absorbed during experiments was subtracted from the total
nitrogen and the remaining number (=total N. before experiment) was calculated as 100.

(2) Toward the end of the experiment, the plants kept in the sugar solution in the
glass house began to suffer. The top of the leaves, dried and turned yellow, but those
kept in the cooler laboratory remained healthy.

(3) Total dry matter of the plants treated with ammonium chloride= 7.901 grams.
» ” ” 7 sodium nitrate= 7.619 ,,
” + Pi ammonium chloride and sugar = 12.393,
sodium nitrate and sugar=10.936__,,

” ” ”

FORMATION OF ASPARAGINE IN PLANTS, 423

After careful washing and drying the entire, plants were

analyzed.
Table XX. In 100 parts of dry matter :—
TEMP. 1-38°C TES
MP. 1-38°C. 515°C
Plants treated with.
= : as
= 2
| | & = wu So
eis Be os | sole silo a |=
fe | Seite | P| ae eS) ee | ee
ois NS ©.) 0S S |coee esac: ee stil a o8
HE, | OS : eS ac Os Ao) 7ey, |]
GS Omnmeemte cs | Sa lca) 2 a fo etna
| < Sas oa |
| < <9)
= == ae — =
Motal nitrogen’ ..........-. 4.28 | 3.64 | 248 | 4.07 | 2.94 | 5.00 | 2.81 | 431 | 2.63
Albuminoid nitrogen...... HOLS || ee! || sey | TOG | Sees || degen |) ase | cGy
|
Asparagine nitrogen’) ... | 040 | 0.72 | 018 | Ogg | 0.27 | 0.82 | o21 | 0.48 | 0.22
|
Nitrogen in nitrates ...... 0.49 | 0.09 O | 0.23 © | 0.76 | 0.14 | 0.16 fo)

As the dry matter had increased very much in those plants
which were treated with sugar, the percentage of the nitrogen
compounds is very much lowered. I calculated therefore the fol-
lowing table to show the relation more clearly :-—

Table XXI. In 100 parts of total nitrogen :—

= ~ TEMP.
EMP. 1-38°C,
Temp. 1-38°C 5-15°C
Plants treated with ;—
= ema = =e
2) | | 2g
| & | eS} a S 3, =
_ | fa bon } iol
elim || Gshe Pee 1 os fo lS ron ls.
ge \22| 6 |e l|s2|2e| 22/58/28
bs 3 s 0) UN |o# (7) S33 ao
Hg, | O's] 2 | Sia | eco tos! su |Sa)iSa
5 O |lS8c|8stn4|B8e]0 ae
| a Ea iS cd
<q nN
Votallmitrogen, Fo). .....+6 100 |100 | 100 | rool2)) r00l?)) rool2)| TOO0"2)} Too =| 100
Albuminoid nitrogen...... 46.3| 38.1] 55.0] 497| 57-8] 392| 53.0| 49.4| 62.4
Asparagine nitrogen ...... Gish UGS) |ieeze3 || 25.0)|) 103 x8.4)) 8-2) nin 3-4
Nitrogen in nitrates ...... DUS |) 22-5) 0 6.5| oO L720} 5-7) Ses 0

(1) Asparagine nitrogen was here determined without previous removal of the bases
by phospho-tungstic acid; therefore the result is somewhat too high.

(2) For the calculation the absorbed nitrogen was subtracted from the total nitrogen;
otherwise a true comparison is impossible,

424 SUZUKI;

It is seen from the above tables that the decrease of aspara-
gine and the transformation of nitrates are enhanced when sugar
is offered, and that on the other hand, an increase of albuminoid
nitrogen takes place, as was to be expected.”

X. Experiments with wheat (Z77tccum sativum).

Young plants 6-10°™ high, were irrigated on the field with
the following solutions :—

a. 0.1% solution of ammonium phosphate.
bo) - ,, sodium nitrate.

Time of experiments :—24 days (Nov. 7th—Dec. Ist).

The temperature was sometimes very cold and fell to
3°C. Total solutions added =about 1000°" in both cases. (two
times irrigated). Rainfall occurred three times during the
experiments.

Table XXII. In 100 parts of dry matter.

Plants treated with

Y ERE War pre S an e ,
Control plants Ammonium phosphate Sodium nitrate
Asparagine nitrogen 0,32 0.42 0.47
Asparagine 1.51 1.07 2.21

Here no noticeable quantity of asparagine accumulated,
very probably because all the conditions for a rapid formation of
proteids were favourable.

Second experiments with wheat.
Young plants 15-20°™ high, grown on the same farm, were

carefully removed, washed, and cultured in the glass house in
glass cylinders, containing :—

a. 500°° of 0.1% solution of urea.

bape, mer os 5 ,, ammonium chloride.
Ge 58 3 0205.05, be ,, ammonium carbonate.
d.= 4,5 ROee x ,, sodium nitrate.

e. distilled water.
Duration of experiment :—11 days (Feb. 3rd—14th).
Temperature :—Miny2°C.; Max. 359°C:

(1) Note on the assimilation of nitrates :—According to Zveud the first product of
assimilation of nitrates in the leaves of Pangium edule, (a tree of Java), is prussic acid.
(Cf. Chem. Zeitg. Feb. 1896). In my numerous experiments, however, on the assimila-
tion of nitrates, the transformation of nitrates to ammonia, asparagine, or proteids, so

quickly proceeded that it was impossible to discover any intermediate product between
nitric acid and ammonia.

FORMATION OF ASPARAGINE IN PLANTS. 425

The solutions were once renewed. The plants were very
healthy, and rapid growth was observed : especially was this the
case with the ammonium chloride plants. The analysis, for which
the entire plants were used, yielded the following results for 100
parts of dry matter :—

Table; XXII

Plants treated with

aS ee
Original Control Ammonium Ammonium Urea Sodium

plants plants _—_ chloride carbonate nitrate
Total nitrogen BaP Byi8 4.48 4.51 4.63 4.60
Albuminoid nitrogen 1.98 1.81 1.64 1.73 1.96 1.99
Asparagine nitrogen 0.19 0.99 1.48 1.33 1.55 1.13
Nitrate nitrogen fo) fo) fo) fo) oO 0.21

Here, a considerable increase of asparagine nitrogen in the
control case was observed. This increase may be chiefly due to
the transformation of other amido-compounds into asparagine,
as there was no nitrate stored up in the plants, and no de-
composition of proteids was observed.

Isolation of asparagine crystals from the plants treated with
ammonium chloride :—1o0 grams of the finely powdered substance
were repeatedly extracted with warm water, and the extract,
after the addition of some tannin and basic lead acetate, was filter-
ed. To the clear filtrate was added an excess of mercuric nitrate
solution, whereby a white flocculent precipitate was formed,
which, after washing, was decomposed with hydrogen sulphide ;
the filtrate from the mercuric sulphide was concentrated on the
water bath, the solution being kept always neutral by the addition

(1) The hot aqueous extract of the wheat prepared with boiling water, gave with
phospho-tungstic acid a turbidity, and upon further addition of nitric acid, a flocculent
precipitate probably due to peptone-like bodies was produced ; but when this extract was
first mixed with basic lead acetate until no more precipitate was obtained, the phospho-
tungstic acid produced in presence of considerable nitric acid, a smaller amount of
pulverulent precipitate, probably due to traces of a base like choline or betain. ‘This
filtrate of the lead precipitate did not give any reaction whatever with caustic potash upon
the addition of a trace of copper sulphate (biuret reaction); therefore no peptone was
present.

Neither ammonia nor urea was stored up as such in the treated plants. A finely
powdered dry sample was extracted with hot absolute alcohol, the alcohol was distilled
off, and the residue after removal of impurities by basic lead acetate, was precipitated
with mercuric nitrate solution, the solution being kept almost neutral, the precipitate was
decomposed by hydrogen sulphide, the filtrate evaporated (keeping it always neutral by
the addition of baryta solution) and the evaporated residue again extracted with warm
absolute alcohol and evaporated again ; hereby xo crystals of Urea, were observed nor
any crystallization upon the addition of nitric acid or oxalic acid.

426 SUZUKI ;

of dilute dammonia. The concentrated solution yielded an abund-
ant quantity of large asparagine crystals which were washed
with water and then with absolute alcohol, and dried over sul-
phuric acid. The amount thus obtained was 0.51 gram,” the an-
alysis proved the crystals to be pure asparagine.

Calculated Observed

Nitrosen, oe. -.. 18.000) era qin
Water of enystallization... 12:0) 9) 12.2%

(lost at 100°C.)
XI. Experiment with barley (Hordeum distichon).
Young barley plants of an average hight of 32° were removed

from field and placed (in the glass house) in the following solu-
tions :—
a. 0.2 9% “urear

b. 96 yeeeand 2 9 sugar.

CG: - ammonium chloride.

d. sn ‘ * and 2 % sugar.
e, 2 96 ssuear

f. distilled water.
Duration of experiment :—7 days (March 26th—April 2nd).
Temperature :—Min. 10°C.; Max 45°C.
After drying, the entire plants® were analyzed with the
following result :—
Table XXIV. In 100 parts of dry matter :—
Plants treated with

F a
Original Control Sugar Urea Ureaand Amm. Amm. chloride

plants _ plants sugar chloride and sugar
Total nitrogen 5.13 5.08 4.60 565 4.98 5:73 5.00
Alb. nitrogen 2.16 1.86 1.47 1.50 1.28 1.60 123
Asp. nitrogen 0.93 1.84 U.94) — r- Si) eet 3.21 2.09
Nitrate nitrogen 0.58 0.37 0.35 0.49 0,24 0.28 0,23

Table XXV._ In 100 parts of total nitrogen :—
Plants treated with

ne a aE te
Original Control Sugar Urea Ureaand Amm. Amm. chloride

plants _ plants sugar chloride and sugar
Total nitrogen 100 100 100 100 100 100 100
Alb, nitrogen 42.3 36.6 32 26.6 25.7 27.9 24.6
Asp. nitrogen 18.1 36.2 A422) | 3351 () eqaee 55:3 41.8
Nitrate nitrogen 11.4 73 7.6 8.6 4.8 4.9 4.6

(1) A moderate quantity of asparagine was left in the mother liquor, which was not
considered.

(2) Toward the end of the experiments some leaves or tips of leaves began to dry
up, owing to the high temperature. Such leaves were, of course, removed,

FORMATION OF ASPARAGINE IN PLANTS. 427

In this table we see that the ratio of albuminoid nitrogen, etc.
is very low, but this does not indicate the decrease of these nitro-
gen compounds. The albuminoid nitrogen is only relatively
lowered, the total nitrogen being considerably increased.

I sought to prove further that asparagine accumulates, even
in presence of much sugar, when some of the other necessary
agents for protein formation are wanting. For this purpose, the
young barley plants of about 32™ > high, were first cultured in the
2% sugar solution. After 7 days, a portion was directly dried
and analyzed, while another portion, was divided into three
equal parts, and cultured in the following solutions :—-

a. 0.2 % solution of urea.

loy * Bs ,, sodium nitrate and 2 % sugar.
CG as fe ,, urea and 2 % sugar.

The plants exposed to ordinary daylight, were after 7
days (March 26th, April 2nd) washed, dried, and analyzed :—
Table XXVI. In 100 parts of dry matter :—

Original®) Unen Sodium nitrate Urea and
plants. and sugar. sugar,
Total nitrogen 4.60 6.09 5.64 5.98
Albuminoid nitrogen 1.47 1.52 1.52 1.58
Asparagine nitrogen ——_1.94 2.97 2.44 3.15
Nitrate nitrogen 0.35 0.24 0.53 0.25

In this case, itis seen that by offering sugar together with
urea, the asparagine nitrogen was increased considerably. By
the preliminary culture for a week in the sugar solution, some of the
conditions for protein formation had been removed, and, I suppose,
it was the sulphates, that were used up. Thus, all the am-
monium compounds had to be stored up as asparagine and could
not be transformed any farther.

SUMMARY AND CONCLUSION.

(1). Asparagine in plants has two sources :—
(a). Itis derived from the decomposition of proteids.
(0). It is a synthetical product of other nitrogenous
compounds :—

(1) The quantitative determination of asparagine by the crystallization method was not
very satisfactory, as some slimy substance prevented a considerable portion from crystal-
lizing.

428 SUZUKI ;

1. Ammonium salts, (and also urea).
2. Nitrates.

(2). Asparagine is formed not only by keeping full-grown
plants in the dark, but also can be formed in full daylight under
certain conditions,

(3). Synthetic formation of asparagine is only possible,
when sugar is present in the plant, and at the same time some
condition for protein formation is wanting. Excess of sugar
prevents the asparagine formation from proteids, but it does
not prevent the synthetical formation of asparagine ; it even sti-
mulates its formation.

(4). Ammonia is never stored upas suchin plants ; it disap-
pears quickly forming innocuous compounds; but when the ne-
cessary amount of sugar is wanting, the ammonia can not be con-
verted and remains as such (experiments with buckwheat) to a
small extent in the plant ; a larger amount is noxious.”

(5). Ammonium salts are generally better than sodium
uttrate for asparagine production.

(6). Among the several ammonium salts, ammonium chlo-
ride is the best, while the ammonium phosphate is always less
favourable for the formation of asparagine ; very probably the
formation of nuclein and new cells are stimulated very much by
phosphates; the asparagine once formed is thus easily trans-
formed into proteids.

Urea is generally better than ammonium salts for asparagine
production (except with barley experiments).

(7). For the conversion of nitrates, a high temperature and
the presence of sugar are necessary, otherwise they remain as
such stored up in the plants for some time.

(8). The conversion of asparagine into proteids is only pos-
sible when all conditions are fulfilled. One of the most es-
sential conditions is of course the presence of sulphates.

(9). In etiolated shoots sodium nitrate can not be con-
verted into asparagine but urea is capable of yielding it.

(10). In eticlated shoots the application of sugar increases
the amount of asparagine, when sodium nitrate or ammonium
salts are offered.

(1) Mr. Aoyama in our laboratory made some experiments which show very clearly
the poisonous action of ammonium salts, when the necessary amount of sugar is not pre-
sent to transform them into asparagine.

FORMATION OF ASPARAGINE IN PLANTS. 429

ANALYDICAL DATA.

For the determination of total nitrogen cited in the fore-
going pages 0.5 or 1.0%" of air dry sample was mixed with 30°
sulphuric acid, 12°" sugar, 2®™° benzoic acid, 18°" mercury
and 0.5"" copper sulphate, and decomposed and distilled as
usual.

For the determination of proteid nitrogen, Stutzer’s method
was used.

For the determination of asparagine nitrogen Sacchsse’s
method was generally used, but in some cases, when the presence
of organic bases was suspected, phospho-tungstic acid was pre-
viously applied.

For the determination of nitrates, Zzemann and Schultze’s
method was used.

HELIANTHUS ANNUUS.

ASPARAGINE NITROGEN.

Dry matter | Baryta() Nitrogen See
Plants in used in water re- found in a ce
grams. _| placed in c.c. gram, ee ae )
I. 4.535 18.8 0.0178 )
Ammonium nitrate ............... 0.39
2. * 18.6 0.0176
j I. 4.485 23.7 0.0224 }
Ammonium chloride ............ 0.50
2. 4 23.4 0.0221 i
: I. 4.550 9.4 0.0089 }
DOGMUMMILATCN ce sneee senile cnias, 0.20
2. 9 9.4 0.0066
TAP Ss 7.0 0.0064
Controliplantsmer.sesys-caeseedes see 0.15
Ze a 6.8 0.0030
I. 4.405 3.2 0,0029
OriginaleplantSienwearsaaceadenes 0.07
Zs 3 3.0 0.0029

(1) 10 ¢.c. of the standard sulphuric acid were equivalent to 48 c.c. baryta solution,
and yielded 0.374 gram barium sulphate. Hence tc.c. of the baryta solution =0.0009447
gram nitrogen.

430

SUZUKI ;

YELLOW LUPINE.
ASPARAGINE NITROGEN.

Dry matter | Baryta Nitrogen es egy
Plants in used in water ()) re- found in |, a eee
grams. |placedinc.c.}| grams. pares:
( I. 1.148 3.8 0,0036
Ammonium phosphate............ 4 0.31
| 2. ” 3-7 0.0035
j I. 1.829 11.6 OOIIO
Utes Sseneaicccn. ste otooe eet eeeee 0.59
(a = 11.4 0.0108
{ I. 1.381 Bez 0.0048
Sodiunnmi (tate <.- cesta a eeeeee 0.36
(ee, 5.2 0.0048
I. 0.919 2.0 0.0019
Controliplantsye-n-era-eca eee 0.20
2. 7 1.9 0.0018
Tis igs 3.8 0.0036
@xiginaliplants) ce--ceeese-.ase eee 0.19
2. 33 3.6 0.0034
MELIA JAPONICA.
ASPARAGINE NITROGEN.
Dry matter Baryta Nitrogen peteece
Plants in used in water (2) re- found in a ee se
grains. placed in c.c. grams. spare
nitrogen.
(I. 4.568 3.0
Ammonium phosphate ......... 0.0067 O.15
7 5s Be
I. 4.368 4.0
Ammonium chloride ............ 0.0082 0.19
2. 3”? 3-7
: | I. 4.373 2.7
Sodium nitrates...................-. 0.0058 0.13
2. ” 2 7 ;
( I. 4.420 1.2
Controliplantss penn-cuseer ee eee 0.0028 0.06
2. ; 1.3
(et 4.073 a
Original ‘plants)sccsse-csse-seeeaeeee 0.0026 0,06
ee 5 ia

(1) 1c.c. of the baryta solution=0.0009447 gram nitrogen.

(2) 10 ¢.c. of the standard sulphuric acid were equivalent to 64¢.c. baryta solution
and yielded 0.572 gram barium sulphate. Hence 1 c.c. baryta solution=0.002148 gram

nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 431

SQUASH (L)

ALBUMINOID NITROGEN,

Plants in Peet ae Baryta water()|Nitrogen found} Percentage of
EAE rain replaced in c.c. in grams. nitrogen.
gram.
Loy Ox 16.7
Ammonium nitrate ... 0.0181 2.58
Z 53 16.9 }
( 1. 0.788 19.2
Sodium nitrate ......... 0.0208 2.53
2. a 19.2
I. 0.694 15.9
Control plants ......... 0.0173 2.49
oo = 16.1

ASPARAGINE NITROGEN.

Percentage of

Plants in Ce ee Soda water 2) |Nitrogen found} nitrogen (=4
Fon) a aeiie replaced. in grams. asparagine ni-
g “ trogen.)
Eee OF7 TK 3.2
Ammonium nitrate ... 0.0140 1.97
2. » 3:3
1. 0.788 2.9
Sodium nitrate ......... 0.0131 1.66
Ze = 3.1 |
I. 0,694 ToL
Control plants ......... 0.c048 0.69
24, 5 1.1

(1) Ioc.c. of the standard sulphuric acid were equivalent to 66.8 ¢.c. baryta solution
and yielded 0.600.603 gram barium sulphate, Hence 1 c.c. of the baryta solution = 0.001094
gram nitrogen.

(2) 10 ¢.c, of the standard sulphuric acid were equivalent to 19.75 ¢.c. soda solution,
and I¢.c. of the soda solution =0,004364 gram nitrogen.

432 SUZUKI ;

SQUASH (IL)

ALBUMINOID NITROGEN.

Plante in ec aE Baryta water/Nitrogen found} Percentage
used in grams, replaced inc.c.| in grams. of nitrogen.
; f I. 0.799 25.5
Ammonium nitrate ... 0.0241 2.99
2. » 25-5
; : (I. O1753 24.9
Sodium nitrate ......... 0.0236 3.11
25.1
12.4
Control plants ......... 2,82
12.5

ASPARAGINE NITROGEN,

Dry matter vin; Percentage of
Plants in (free from ash) cathe vee el Sean nitrogen (3 asp.
used in grams, | "P" i alters nitrogen.
F : I, 0.799 II.I 0.0105
Ammonium nitrate ... 1.28
2s os 10,7 0.0101
I. 0.752 9.4 0.0089
Sodium nitrate ......... 1.17
2. 5 9.1 0.0086
I, O.411 3.6 0.0034.
Control plants ......... 0,82
2. ” 35 0.0033

(1) 1¢.c. of the baryta solution=0.0009447 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 433

POTATO (L)

ASPARAGINE NITROGEN.

ele Percentage of
c Dry matter |Baryta water(2/Nitrogen found] - g
Plants in used in grams. | replaced in c.c.| in grams. Was
c ‘ I. 6.955 55.8 0,0527
Ammonium nitrate ... 0.76
2. ” 55-4 0.0523
I. 5.981 30.8 0.0290
Sodium nitrate ......... 0.49
2, a Br.2 0.0295
I. 7.049 27.8 0.0263
Control plants ......... 0.38
2. as 28.4 0.0267
2 I. 2.097 2.5 0.0024.
Original] plant, roots... O.1I
2, 3 2.3 0.0022
2 I. 2.558 2.6 0,0025
Original plant, stems... 0.10
By 3 2G 0.0026
POTATO. (II).

ASPARAGINE NITROGEN,

Dry matter

Plants in moot

Grams.
4.492

1.797

Ammonium phosphate

1.156

Sodium Nitrate .........

Control plants .........

(1) 1¢.c. baryta solution =0,0009447 gram nitrogen,
(2) 1¢.c, baryta solution =0,0009447 gram nitrogen.

Baryta water(?)
replaced.

Percentage of

ak Se nitrogen (4 asp.
‘ nitrogen.)

Gram.
0,0082

0.19
0.0034
0.0032

0.27
0.0030
0.0065

0.28
0.0066

434 ; SUZUKI ;

POTATO SHOOTS. (1).

ALBUMINOID NITROGEN.

Planisan Dry matter |Baryta water() Nitrogen Percentage of

used, replaced. found. nitrogen.
Gram. aCe Gram,

Wier & Sates caneocacouannden 0.834. : 0.0159 1.9!

Sodium nitrate ......... 1.288 : 0.0231 1.79

Urea and Sugar......... 0.770 : 0.0162 2.11

Sod. nitrate and Sugar 0.635 X 0.0116 1.80

Control plants ......... 0.641 : 0.0122 1.91

Original plants ......... 0.645 : 0.0141 2.18

ASPARAGINE NITROGEN.

Percentage of

Si ( * E
Dry matter |Baryta water() Nitrogen nitrogen (J asp.

Plants in

used. replaced. found. nitrogen.)

Gram. Gilc: Gram.
WOR Car Sele ncurcs aoceennte 0.834 22 0.0069 0.82
Sodium nitrate ......... 1.288 2.0 0.0062 0.48
Urea and Sugar......... 0.770 1.7 0,0053 0.69
Sod. nitrate and Sugar 0.635 1.2 0.0037 0.59
Control plants ......... 0.641 se) 0.0031 0.49
Original plants ......... 0.645 0.7 0.0022 0.34

(1) 5 ¢.¢c. of the standard sulphuric acid were equivalent to 22 ¢.c. baryta solution and
yielded 0.5695 gram barium sulphate. Hence 1Ic.c. baryta solution=0.0031245 gram
nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 435

POTATO SHOOTS. (II).
TOTAL NITROGEN.

Plants i Dry matter |Baryta water™)| Nitrogen Percentage of
oe used. replaced. found. nitrogen,
Gram. Cues Gram,
( I. 0.456 4.0
Sogarineccecacecsssce 0.0125 2.74
| 2. +, 4.0
) I. 0.454 5-3
Weaeermeas..nsetedssosscss 0.0169 oui
2 oy 55
I. 0.440 4.8
Sodium nitrate ......... 0.0153 3.48
2 3 5.0
Sodium nitrate and ( I. 0.472 46 |
SUGATceecaresccisncw eos 4 0.0144 3.04
| 7. Pe 4.6 j
I. 0.447 4.7 |
Control plants ......... - 0.0147 3.29
Po P| 4.8 \
I. 0.478 4.7 }
Original] plants ......... - 0.0150 3.14
2. ” 4.9
ALBUMINOID NITROGEN.
- Dry matter |Baryta water() Nitrogen Percentage of
ENS oe. used. replaced. found. nitrogen.
Gram, cc. Gram.
I. 0.912 4.2
Sugary. scnescccsess-necens 0.0132 1.45
(2. ” 4.4
I. 0.908 ise)
NY LCA ys ce aiitace-weseasjroness 0.0179 1.97
| 2. ” 5-7
\ I. 0880 5.2
Sodium nitrate ......... 0.0167 1.89
(2a, 5.4
Sodium nitrate and|({1I. 0.944 5.3
SLUCEh dl cenebansesenapadee 0.0170 1.80
2. ” 5.4
I. 0.894 5.3
Control plants ......... 0.0170 1.86
3- ” 5.3
> j I. 0.956 5.4
Original plants ......... 0.0173 1.81
2. » 5.6

(1) 1¢.c. baryta solution =0,0031245 gram nitrogen.

436 SUZUKI ;

ASPARAGINE NITROGEN.
— een

‘ Dry matter |Baryta water Nitrogen found Percentage et
HBOS used in grams. | replaced in cc.| in grams, _|Togen (3 asp.
nitrogen.)

I. 1.824 22

SEEMS stisggacenqousaga00c0 0.0069 0.38
2 es 2.2
I. 1.816 Bin

Urea’ ain ccsstencotes ses as 0.0162 0.90
2 5 53
I. 1.760 2.1

Sodium nitrate ......... 0.0069 0.39
2 as 2.2
Sodium nitrate and I. 1.888 1.6

SUGAM) corccteseaiescesee 0.0053 0.28
2. ki 1.8
I. 1.788 2.0

Control plants ®......... } 0.0062 0.35
2, 3 2.1
I. O12 1.7

Original plants ......... 0.0053 0.28
2 3 et

NITRATE NITROGEN.

Height of Volume of| Nitrogen

: Dry matter ‘Tempera- Bi - | Percentage
Plants in : barometer = NO gas in| found in :
in grams. | 5, inches, | tre C ae of nitrogen,

0.880 0.21

17
18 2.6

Sodium nitrate... 30.12

Sodium nitrate
and sugar......

0.16

0.944

”

(1) 1¢.c. baryta solution =0,0031245 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS, 437

HOLESIA HISPIDUM.

ASPARAGINE NITROGEN,

F Percentage of
. Dry matter | Baryta water) Nitrogen = 8
Plants in used. replaced, found. ieee ee
Gram. Oe Gram.

I. 3.664 3.0 |

Ammonium phosphate 0.0028 0.08
2s 3.0 j

Ammonium phosphate Pa sea 9 0.0019 0.05

angisuganiens. earn. lain ues 2.1

\ I. 3.668 4.2 )

(Wreag mars .posmicnaressene 0.0042 O11
| 2. ” 45
I. 1.790 1.0 )

Urea and sugar......... 0.0010 0.06
2. 9 1.2 {
—een( I. 3.692 3.0 )

Control plants ......... - 0,0028 0.08
a ess 3.0 |
{ Te) 2.60% 15 )

Original plants ......... - 0.0015 0.06
| 2s 1.6 |

BUCKWHEAT. (1.)

DETERMINATION OF AMMONIA.

One gram of air dry matter of the ammonium nitrate plant
(lower stems and roots) was extracted with warm water and to
the filtrate, to which so much sodium bicarbonate added as
necessary to render the liquid slightly alkaline, yielded on
distillation 0.0017 gram nitrogen (=1.8cc. baryta solution”)
=0.17% nitrogen.

Air dry samples were mixed with some caustic lime and
moistened with water. The mixture was kept under a bell jar.
The ammonia liberated was absorbed by a known quantity of
standard sulphuric acid ; thus I obtained the following results :—

I gram of air dry sample of the control plants yielded
0.0004 gram nitrogen (=o.4cc. baryta solution™”)=0.04.% nitrogen.

(I) 1¢.c. of the baryta solution = 0.0009447 gram nitrogen.

438 SUZUKI ;

I gram of air dry sample of the plants in ammonium nitrate
yielded
0.0019 gram nitrogen (=2.0cc. baryta solution”) =0.19% nitrogen.
I gram of air dry sample of the plants in sodium nitrate yielded
0.0008 gram nitrogen (=0.8cc. baryta solution) =0.08%
nitrogen.©

BUCKWHEAT (IL)

ASPARAGINE NITROGEN.

Percentage of

| |
E |Baryta water) Ni :
Fresh sample |Baryta water@) Nitrogen found nitrogen (=4

Plants in

used in grams.| replacedincc.} in grams. asp. nitrogen.)
Ammonium chloride... 5.0 : 0.0051 0.10
Ammonium nitrate ... 5-0 : 0.0030 0.06
Sodium nitrate ......... 5.0 4 0.0015 0.03
Control plants ......... 5.0 é 0.CO13 0.03
AMMONIACAL NITROGEN.
: Fresh sample |Baryta water()|Nitrogen found] Percentage of
Plants in 3 : ; :
used in grams. | replaced in cc. in grams. nitrogen.)
Ammonium chloride... 7:45 6.0 0.0057 0.08
Ammonium nitrate’... 6.20 Qu 0.0026 0.04

(1) Ic.c. of the baryta solution=0.0009447 gram nitrogen.

(2) There are doubtless also traces of asparagine decomposed by calcium
hydrate at the ordinary temperature; the small quantities of ammonia obtained from the
control plants and the plants in sodium nitrate seem to be due to this source. Control
test with Nessler’s reagent (added to the cold extracts of the control plants and plants in
sodium nitrate) lead me to this inference.

FORMATION OF ASPARAGINE IN PLANTS. 439

BUCKWHEAT (IIL)

ASPARAGINE NITROGEN.

: a f
Phat in Dry matter Baryta water Nitrogen found Heer aa
4 used in grams. replaced in cc. in grams. asp. nitr ogen.)
Ammonium chloride |
Ke)

AiG! FRPEERS A paapndoe f 1.840 0.95 Ope09. 205
Ammonium nitrate ... 1.910 3-9 0.0037 0.19
Ammonium nitrate and |

SUGAR rani eras ( 1.864 12 ono a)
Ammonium carbonate. 1.902 48 0.0046 0.24
Ammonium carbonate 1.826 | aC 0.0025 0.14

Euaval Gelecto: mecuosbaabee ; i
Ammonium phosphate. 1.852 2.5 0.0024 0.13
Ammonium phosphate

andiSUCalNe.csef..-5- 1.755 29 CPEO? 295

BRASSICA (W/KAWASHIMA.)

ASPARAGINE NITROGEN.

d Percentage of
nitrogen (} asp.
nitrogen.)

Dry matter

Baryta water l)|Nitrogen foun
used in grams,

Plants in : :
replaced incc,| in grams.

Ammonium phosphate 0.866 0,0024 027
Sodium nitrate ......... 0.885 0.0037 0.41
Control plants ......... 0,864 0.0037 0.42

(1) Ice, baryta solution = 0.0009447 gram nitrogen.

440 SUZUKI;

BRASSICA, SWEDISH TURNIP. (1)
ASPARAGINE NITROGEN.

Percentage of

: Dry matter |Baryta water() Nitrogen nitrogen
Plants in i
used, replaced, found. (% asparagine
nitrogen.)
Gram. Cc Gram.
L710 68.8

Ammonium Nitrate ... | 0.0653 0.85

2. PA 69.4

Ammonium carbonate

Teton 74.2
0.0608 0.85

2s am 74.6

2.3 78.5
I. 9.814 78.2
Be 3 78.6

Control plants

0.0741 0.75

Original plants

0.0123 0.32

} | I. 7.257 78.3
Sodium nitrate ......... 0.0741 1.02

| I. 3.774 12.8

2. 13.0

BRASSIG@ OW EDISH TURNIP (1)
ASPARAGINE NITROGEN.

Percentage of

- Dry matter |Baryta water() Nitrogen nitrogen
anc used, replaced. found. (4 asparagine
nitrogen, )
Gram, CAG: Gram,

(I. 3.591 23.6

Ammonium chloride 0.0225 0.63
Psy 24-0
I, 2,968 18.1

Ammonium phosphate 0,0170 0.57
2. gt 17.9
I; 31673 21,3

Sodium nitrate ......... 0.0203 0.55
25 aie; 27
I. 3.669 28.3

Control plants ......... 0,0269 0.73
= 2: ess 28.7
I. 3.651 10.4

Originaf plants ......... 0.0100 0.27
2 10.6

i LL

(1) 1e.c. baryta solution =0,0099447 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 441

NITRATE NITROGEN,

. : Percentage
Dry matter y ‘ Tempera- | Volume of | Nitrogen 8
used, {S'S Mirae, NO gas, | found. seed
nitrogen,
Gram. Inch. ec: CuGs Gram.
I. 0.913 30.36 9 11.9
Control plants ... 0.0076 0,83
2. ” ” ” 12.I
I. 0.917 i * 6.0
Original plants... 0.0038 0.41
2. ” ” ” 6.0

BRASSICA. (TAINA). (1)

ASPARAGINE NITROGEN,

Percentage of
Plantsin Dry matter |Baryta water‘) Nitrogen nitrogen
sr: used. replaced. found. (% asparagine
nitrogen.)
Gram. Gics Gram,
Ammonium phosphate 1.840 2,0 0,0043 0.23
Sodium nitrate ......... 1.830 1.8 0,0039 0.21
Control plants ......... 1.858 7, 0.0037 0.20

BRASSICA. (TAINA). (II)

I. Nitrogen determination of ammonium chloride solution
applied to the plant (before experiments). 5°° of the ammonium
chloride solution were diluted with water to 250°%, and from
this 20°" and 100°" were distilled after addition of soda solu-
tions.

I. 20°° yielded 0.0160 gram nitrogen (=16.9°° baryta
solution”) =0.08% nitrogen.

2. 100°° yielded 0.0792 gram nitrogen (=83.8°° baryta
solution) =0.08% nitrogen.

This calculated to the original solution gives

I. 3-996
2. 3.96%. Average 3.98%.

(1) Icac, baryta solution =0,0009447 gram nitrogen.

442 SUZUKI ;

Hence 1°” of the ammonium chloride solution contains 0.0398
gram nitrogen.

15°° of the ammonium chloride solution contains 0.5963 gram
nitrogen,

II. Nitrogen determination of the ammonium chloride solu-
tion that had remained unabsorbed, (after experiment). Total
unabsorbed solution were filled up to 500°°, from which 20°%
and 30° were distilled as above.

I. 20°* yielded 016220 gram nitrogen “(=23'3"— Jbanyta
solution) =0.1% nitrogen.

2. 30°° yielded 0.0330 gram nitrogen (=34.9"" baryta
solution) =0.19% nitrogen,
which, calculated to the original solution, give

if 3.67%
2 3.66%. Average 3.67% nitrogen.

Therefore, total nitrogen remained unabsorbed=o0.5501 gram.
Total nitrogen originally applied =0.5963 gram.
Total nitrogen absorbed =0.0462 gram.
III. Nitrogen determination of sodium nitrate solution ap-
plied to the plant (before experiment.)
10° of the sodium nitrate solution were diluted up to 100°,
from which each 10°* were taken for analysis.
I. 10°° yielded 0.0150 gram nitrogen (temp. 6 °C., pressure
29.84 inches ; NO=24.5°“).
=1.50% or 15.0% of original solution.
2. 10°° yielded 0.0150 gram nitrogen (temp. 6.°C., pressure
29.84 inches ; NO=24.5°").
Hence 30°° of the sodium nitrate solution contain 0.4497
gram nitrogen.
VI. Determination of unabsorbed nitrogen in the sodium
nitrate solution.
Total unabsorbed solution were concentrated to 1000°* from
which 50°* were analyzed.
I. 50°° yielded 0.0203 grams nitrogen (temp. 14.5°C., pres-
sure 30.18 inches ; NO =33.8°* ).
2. 50°° yielded 0.0203 grams nitrogen (temp. 14.5°C., pres-
sure 30,18 inches; NO=32%6-~).
Total nitrogen remained unabsorbed =0.4060 gram.
Total nitrogen originally applied =0.4497 gram.
Therefore, total nitrogen absorbed by plants=0.0437 gram.

FORMATION OF ASPARAGINE IN PLANTS, 443

TOTAL NITROGEN.

Plants i Dry matter |Baryta water() Nitrogen Percentage of
enon used. replaced. found, nitrogen,
Gram. Cae. Gram.
I. 0.472 10.5
Ammonium chloride... 0.0228 4.83
0p 5 10.6
2 : j I. 0.454 10.0
Sodium nitrate ......... r 0,0217 4.78
| 2s 10.2 \
T. 0.455 94 )
Control plants ......... 0.0202 4.44
2 ” 94 j
. I. 0.462 9.8
Orginal plants ......... 0.0213 4.60
2 es 10.0
ALBUMINOID NITROGEN,
A
; : Dry matter | Baryta water(?) Nitrogen Percentage of
Plants in used, replaced. found. nitrogen.
Gram CHC: Gram.
I. 1.888 16.1
Ammonium chloride... | - 0.0350 1.86
| Zs 16.5
; ; I. 0.908 5.6 )
Sodium nitrate ......... 0.0122 1.35
| 2 ~ 5.8 j
I. ©.g10 6.9
Control plants ......... 0.0148 1.63
Pe 9 6.9 )
oh I. 0.924 7.0 }
Original plants ......... > 0.0153 165
2h Ve 7.2

(1) 1c. baryta solution=0 002148 gram nitrogen.

444

SUZUKI ;

ASPARAGINE NITROGEN.

Percentage of

F Dry matter | Baryta water() Nitrogen nitrogen
Eapisnm used, replaced. found, (} asparagine
nitrogen.)
Gram, G:C; Gram.
I. 1.888 3.8
Ammonium chloride... 0.0082 0.43
2. ” 3.8 /
I. 1.816 4.1
Sodium nitrate ......... 0.0086 0.47
20. Gs 3-9
I. 1.820 3-1
Control plants ......... 0.0064 0.35
2 2.9
1. 1.848 1.2
Original plants ......... 0.0026 0.14
2) ae 1.2
NITRATE NITROGEN.
Plants Dry matter Presente. Tempera- Volume of| Nitrogen Rene ee
used. ture. NO. gas. | found. nitrogen
Gram Inch. cc C.\G: Gram.
( I. 0.944 20.7 14.5 10.0
Amm, chloride... 0.0058 0.62
l 2. P ” ” 9.8
I. 0.908 - sf 133 i
Sodium nitrate... 0.0080 0.88
2. ” ” 7 13.7
\ I. 0.910 35 16. 5.4 )
Control plants ... - 0.0032 0.36
| 2. ” ” > 5 6
Bs I, 0.924 es ns 16.3 )
Original plants .. 0.0096 1.04
2. ” ” ” 16 4 f

(1) 1e.c. baryta solution= 0.002148 grain nitrogen,

FORMATION OF ASPARAGINE IN PLANTS. 445

BRASSICA CAMPESTRIS VAR. TAINA. (III.)

I, Examination of ammonium chloride solution that remained
unabsorbed. The total solutions were diluted to 500°", of
which 200"* and 100°* were analyzed.

(a). 200°* of the ammonium chloride solution yielded
0.0627 gram nitrogen (=66.3°" baryta solution).

Hence total nitrogen in total solution=0.1568 gram,

(b). 100%° of the ammonium chloride and sugar solution yielded
0.0321 gram nitrogen (=34°" baryta solution).

Hence total nitrogen in total solution=0.1606 gram.
Total nitrogen originally applied =0.1988 gram.

Therefore, 0.1988 —0.1568=0.042 gram nitrogen was absorbed by
ammonium chloride plants.

0.1958 — 0.1606=0.0382 gram nitrogen was absorbed
by ammonium chloride and sugar plants.

II. Examination of sodium nitrate solution that remained
unabsorbed. The total solutions were concentrated to 100°,
from which 20 and 10° were analyzed.

(a). 20° of sodium nitrate yielded
0.0218 gram (temp. 15°C., pressure 29.66 inch., vol. of
INO 3614552),
Total nitrogen in total solution=0.1090 gram.

(b). 10°° of sodium nitrate and sugar solution yielded
0.0113 gram nitrogen (temp. 15°C ; pressure 30 inches ; vol.
Of NO! 1825¢°°):

Total nitrogen in total solution=o0,1126 gram.
Total nitrogen originally applied =0.1499 gram nitrogen.

0.1499—0.1090=0.0409 gram nitrogen was absorbed by
sodium nitrate plants.

0.1499 —0.1126=0.0373 gram nitrogen was absorbed by
sodium nitrate and sugar plants.

446 SUZUKI ;

TOTAL NITROGEN,

- | Dry matter Baryta water() Nitrogen Percentage of
Plants la | used, replaced. found nitrogen,
|
Gram. (So Gram,
I. 0.477 5.6
Sugar e.cocssscaccoetseeses 0.0118 2.48
| 2. ” 5-4
| j I. 0.465 8.7
Ammonium chloride... 0.0189 4.07
| 2° a 8.9
Ammonium chloride I. 0.482 6.5
ANGISUGAT teeter ae 0.0142 2,94
2 % :

0 0129 2.81

Sodium nitrate and |( 1. 0.459 | 6.0
GUIEANS . sshascooodtp00a00¢

| (I. 0447 | 10.4
Sodium nitrate ... . ... | 0.0223 5 00

(1) 1¢.c. baryta solution =0.co2148 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 447

ALBUMINOID NITROGEN,

. Dry matter |Baryta water‘! ) Nitrogen Percentage of
Fuse used. replaced. found. nitrogen.
Gram. ene: Gram. |
I. 0.954 6.0
SUGAN ee cccaees enc seceeecs | 2 | 0.0131 1.37
| 2 A 6.2
Ammonium chloride plea i 0.016 1.76
Cree i | 0103 7
2. oD 7 7
Ammonium chloride \ I. 0.964 6.8
and sugar ............ 0.0146 1.52
2 pee 7.0
| I. 0.894 73 | }
Sodium nitrate ......... | 0.0157 1.75
2 ” 74 |
Sodium nitrate and { I. O9Ig 56 |)
SUQAE Ct cer csncnowacsns | j 0.0120 1.31
(2 » ens!
Control pl 5 ee e |
Dias see ace 2 0 0133 1.43
2 + e
- \ I. 0.964 go |
Original plants ......... 0.0193 1.98
(% ” 9.0
— eee ee

(1) 1¢¢. baryta solution=0.002148 gram nitrogen.

448 SUZUKI ;

ASPARAGINE NITROGEN.

= | hese Percentage of
Plantsun Dry a Bary ay eee ae Seon
used, rep aced, ound, asp. nitrogen).
Gram, cxc Gram.
a | I. 1,908 0.7 |
(or Vers pondoeeaanaadconede i ( O.COI7 0.09
2 ¥ 0.9
I. 1.860 4.2 |
Ammanium chloride... 0.0092 0.50
a: a4
Ammonium chloride | I, 1.928 1.2 F
ANGISUS AT elessesceeee | ( 0.0026 O13
2s ; 1.2
eae f I. 1.788 3:3 )
Sodium nitrate ......... | 0.0073 0.41
2. 5 3-4
Sodium nitrate and { I. 1.838 0.8 )
SOG al io. Secceecssen se | ( 0.0019 0.11
2a 1.0
| I. 1.840 Bat )
Control plants ......... i ( 0 0067 0.36
20 3-1
— | I. 1.928 1.7 ]
Original plants ......... ] f 0.0039 Chale
23 an 1.9
NITRATE NITROGEN.
Plantsan Dry matter Preseare: Tempera- Volume of} Nitrogen ee
used. ture. NO gas. | found. nitrogen
Gram. Inch. oc: Cics Gram.
Sugar’ sicpoceecs 0.954 29.84 | 6 fo) fe) fo)
Amm. chloride... | 0.930 59 7 a5 0.0021 0.23
Amm. chloride
and sugar...... | one4 a 7 % a *
Sodium nitrate... | 0.894 29.7 16 11.5 0.0068 0.76
Sodi itrat
adieugar ae t 0.919 30.17 13 2.1 0.0013 0.14
Control plants ... 0.920 29.84 6 1.4 0,0009 0.09
Original plants... 0.964 i | 7 Tel 0.0047 0.49

(1) 1¢.c. baryta solution=0,002148 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS. 449

BRASSICA. (TAINA). (IV.)
TOTAL NITROGEN.
Dry matter |Baryta water( Nitrogen Percentage of
used. replaced. found. nitrogen,
Gram. Cc, C. Gram.
{ I. O491 6.1 |
Plants in sugar ......... , - 0.0129 2.63
| 2. ” 5 9 |
| I. 0.489 9.8 )
Control plants ......... r 0.0211 4.31
| 2. » 99

ALBUMINOID NITROGEN.

Plants in sugar

Control plants

Plants in suger

Control plants

Dry matter
used.

Gram.

(I. 0.981
ye

{ I. 0.978

Weiss ot 5

Baryta water() Nitrogen Percentage of
replaced. found. nitrogen.
cic: Gram.
7.6 )
0.0161 1.64
a)
9-7 l
0.0208 2.13
a7 |

ASPARAGINE NITROGEN.

Percentage of

Dry matter |Baryta water) Nitrogen anes
used. replaced. found. waste Ce (OSCE 2
nitrogen),
Gram, 7 Gram.
I. 1.962 1.0 )
1 0.0021 0.11
2 > I.I f
I. 1.956 2.1
| 0.0047 0.24
DP oe 2.3

(1) 1¢.¢. baryta solution =0,002148 gram nitrogen.

450 ; SUZUTI ;

NITRATE NITROGEN,

7 ee Percentage
Dry matter Tempera- | Volume of} Nitrogen 8
used. | SaESSHTE. ture. | NO.gas. | found. me
nitrogen,
Gram. Inch. °c Cac. Gram,
I. 0.981 °
Plants in sugar...
2. 5 fo)
( I. 0.978 20.7 16 2.7 )
Control plants ... | 0.0016 0.16
| 2. ” ” ” 2.8 (
WHEAT (I).
ASPARGINE NITROGEN.
- Percentage of
- Dry matter |Baryta water) Nitrogen :
HES aa replace found, Poneto
nitrogen).
Gram. CAC. Gram.
1. 1.866 ey
Ammonium phosphate 0.0039 0.21
2) es 1.9
I. 1.836 2.0
Sodium nitrate ......... 0.0045 0.24
Dy 2.1
I. 1.890 1.4
Controliplants) eas... 4- 0,0030 0.16
2. 65 1.4

(1) 1 ¢.c. baryta solution =0.002148 gram nitrogen.

FORMATION OF ASPARAGINE IN PLANTS.

WHEAT (1).

TOTAL NITROGEN,

451

. Dry matter |Baryta water()
Plants in cea! replaced.
|
Gram. Crc.
I. 0.472 9.7
Ammonium chloride...
Zs 3 10.1
\ I. 0.467 9.7
Ammonium carbonate
ido. 10,0
i I. 0.465 10.1
WEAN petrasessanetrc'ee fete
| 2. 3 10.0
I, 0.465 10.1
Sodium nitrate .........
2. #) 98
j I. 0.476 8.4
Control plants .........
| Ze 3 8.2
I. 0.484 8.5
Original plants .........
oy is 8.3

Nitrogen
found.

Gram.
0.0206
0.0217
0.0206
0.0215
0.0217
0,0215
0.0217
0.0211
0.0180
0.0176
0.0183

0.0178

Percentage of

ST ea Cees ieee

nitrogen.

4.48

451

4.63

4.60

3-73

3:72

Plants in

ALBUMINOID NITROGEN,

Dry matter

Ammonium chloride |} -

Ammonium carbonate

Sodium nitrate .........

Control plants .........

Original plants .........

used,

Gram.

| I. 0.944
2. ”

( I. 0933
| 2. ”

I. 0.930
f 2. es

| I. 0.929
2s -

I. 0.952
4, <

{ I. 0.968
(aes ie

Baryta water)
replaced,

Gc;
7.0
7.2
7-4
75
8.4
8.6
8.5
8.7
8.0
8.0
9.0
8.9

Nitrogen Percentage of
found, nitrogen.
Gram,

0.0151

1.64
0.0155
0 0160

1.73
0.0161
0.0180

1.96
0.0185
0.0183

1.99
0.0187
0.0172 )

1.81
0.0172 /
0.0192

1.98
0.0190

(1) 1c. of the baryta solution = 0,002148 gram nitrogen.

452

SUZUKI ;

ASPARAGINE NITROGEN.

- Percentage of
; F Dry matter |Baryta water(!) Nitrogen F 2
PEAS used, replaced. found. nitrogen (f Asp:
nitrogen).
Gram. Cx: Gram,
{ I. 1.888 6.6
Ammonium chloride | - 0.0140 0.74
| 2 5) 6.4
I. 1.866 5.6 l
Ammonium carbonate | - 0.0125 0.67
liar ie 59 j
I. 1.860 66 )
OS g=A ae Namen ben eneet Gacdar 0.0144 0.77
2. “5 6.7 f
(1. 1,858 5.0
Sodium nitrate ......... | 0.0105 0.57
2. ” 4.7
I. 1.904 45
Control plants ......... 0.0095 0.50
2. ” 4.3
PI. 1.936 1.2
Original plants ......... 0.0028 0.14
2 - 1.4 )
NITRATES NITROGEN.
: Dry matter} Height of | Tempera- | Volume of| Nitrogen REISE AES
ES used barometer, ture NO. gas found e
: : eee: : nitrogen,
Gram, (Inch.) te. Cuics Gram.
I. 0.929 30.2 15°C 3.2
Sodium nitrate... 0,0020 | 0.21°/,
2. 0.929 30.2 15°c 3.4

(I) 1e¢.c. baryta solution=o 002148 gram nitrogen,

FORMATION OF ASPARAGINE IN PLANTS. 453

BARLEY (I).

TOTAL NITROGEN.

: Dry matter |Baryta water?) Nitrogen — | Percentage of
BENS used. replaced. found. nitrogen,
Gram. Gs & Gram,
I, 0.467 11.5
SIRVEETT cancogacednsocecooces 0.0215 4.60
Zs 3 11.5
I. 0.460 13.6
Wea ree as specie seen cone 0.0260 5.65
By, FF 14.0
I. 0.46 12.4
Urea and sugar........, : 0.0232 4.98
2. 55 12.2
( I. 0.470 14.1 }
Ammonium chloride 0.0270 5-73
| 2 ” 14.4 j
Ammonium chloride |(1. 0.459 12.1
and sugar .....,...... 0.0230 5.00
2 55 12.3
{ I. 0.4705 12.7
Control plants ......... 0.0239 5.10
| 2 Bs 12.7
I. 0.477 12.9
Original plants ......... 0.0245 5.13
2 5 13.1

(1) 5¢¢. standard sulphuric acid were equivalent to 36.5 ¢.c. baryta solution and
yielded 0.5695 gram barium sulphate. Hencetrc.c, baryta solution=0.001883 gram
nitrogen.

454

ALBUMINOID NITROGEN.

SUZUKI ;

- Dry matter |Baryta water(1)
FERS used, replaced.
Gram Gic
I. 0.467 Ze
S)TEEV co nadbobagoI0031 000000
2. 3 Pape
I. 0.460 2a
MITCa cacecneececeatmosenes :
Py, + 2.3
I. 0.465 2.0
Urea and sugar.........
2. % 2.2
I, 0.470 De
Ammonium chloride
2. + 2.4
Ammonium chloride |(1I. 0.459 1.8
and sugar ......... 300
(2% 5. 1.8
I. 0.471 277
Control plants ....,....
@, + 2.9
ae | I. 0.954 6.5
Original plants ..... ...
lao 6.6

(1) tcc. of the baryta solution =0,0031245 gram nitrogen.

— — —_—_—o —~’” —o —~ —_os

Nitrogen
found,

Gram.

0.0069

0.0069

0.0065

0.0075

0.0056

0.0087

0.0206

Percentage of
nitrogen.

1.47

1.50

1.39

1.60

1.23

1.86

2.16

FORMATION OF ASPARAGINE IN PLANTS. 455

ASPARAGINE NITROGEN.

“2g Percentage of
: Dry matter |Baryta water‘) Nitrogen z
Plants in used, replaced. found. paa Aa
Gram. Cyc: Gram,
I. 0.634 4.7
SLEVEZN 2 GeucuncontedtRenoone 0.0090 0.97
2. ” 49
I. 0.920 4.6
Wirteaterestesseserecess cc 0.0087 0.95
2. ” 4.7
{ I. 0.930 5.2
Urea and sugar......... 0.0100 1.08
le 3 5.4
I. 0.940 8.0 )
Ammonium chloride 0.0151 1.60
2. rs 8.1 \
Ammonium chloride | (1. 0.918 5.0
ADGsSUGar ssesc<ce.cs 0.0096 1,05
2. 33 5.2
| I. 0.941 4.6
Control plants ......... 0.0087 0.92
| 2 x 46
I. 0.954 2.2
Original plants ......... 0.0044 0.46
2. ” 2.4 |

Dry matter

Plants in weed)
Gram.
Sheree te epecmemenes 0.934
Wreaitteacccser.-<s 0.920
Urea and sugar 0.400
Amm., chloride... 0.940
Amm. chloride
and sugar...... t Bone
Control plants ... 0.941
Original plants... 0.954

NITRATE NITROGEN.

Height of | ‘!'empera-

Inch.
29.8

29.9
29.8
30.12
30.12
30.12

30.12

Barometer. ture.

oC
18

16
18
18
19
18

16

Volume of | Nitrogen | Percentage
NO gas.

c.c.

5-5
7-7
1.6
4.4
3-5
5.8

9.2

found. _ jof nitrogen,

Gram.

0.0032 0.35
0.0045 0.49
0.0009 0.24
0.0026 0.28
0.0021 0.23
0.0034. 0.37
0.0055 0.58

(1) 1ec. baryta solution=0,001883 gram nitrogen,

456

SUZUKI ;

BARLEY (1)

TOTAL NITROGEN,

Plants on Dry matter /Baryta water() Nitrogen Percentage of
4 used. replaced. found. nitrogen.
Gram. Cacs Gram,
( I. 0.474 9.2
Urea ieacarsece espe 0.0291 6.10
| 2. ” 9.4
I. 0.474 8.6
Sodium nitrate ......... 0.0269 5.64
2. 3 8.6
I. 0.476 9.0
Urea and sugar......... 0.0284 5-98
2s 5 9.2
i I. 0.467 6.8
Original plants ......... 0.0215 4.60
2. x 7.0
ALBUMINOID NITROGEN.
: Dry matter | Baryta water) Nitrogen Percentage of
sens used. replaced. found. nitrogen.
Gram. CC: Gram,
I. 0.474 2.3
Wreamnrce sincera 0.0072 1.52
2 55 2.3
I. 0474 2.2
Sodium nitrate ......... 0.0072 1.52
2. 7 2.4
I. 0.476 2.4
Urea and sugar......... 0.0075 1.58
25 24 Ee)
I. 0.467 2.1
Original plants ......... 0.0069 1.47
2 3 2.3

(1) 1¢.c. baryta solution=0,0031245 gram nitrogen.

tm

FORMATION OF ASPARAGINE IN PLANTS. 457

ASPARAGINE NITROGEN.

: Percentage of
- Dry matter |Baryta water) Nitrogen E
EUS en | eticed eran Dau Opens (a asP:
nitrogen.)
Gram, CHC: Gram,
I. 0.947 4.5
(WIKGZ.. “issoehasoossdaecnoans 0.0141 1.49
2. FF 4.6
\ I. 0.948 | 3.5 i)
Sodium nitrate ......... | - 0.0112 1.19
ae P53 CS
I. 0.952 4.1 | )
Urea and sugar......... - 0.0131 1.38
2) A as |)
: I. 0.934 2.9
Original plants ......... 0.0090 0.97
2. 3 2.9

NITRATES NITROGEN.

Planiein Dry matter) Height of} Tempera- | Volume of | Nitrogen Peceneee
used. Barometer.| ture. NO gas. found ns
nitrogen.

Gram. Inch. MG CHC: Gram.

MWiedase. cee racceses 0.947 29.8 18 3.8 0.002233 0.24

Sodium nitrate... 0.948 30.12 16 8.4 0.005023 0.53

Urea and sugar 0.952 29.8 18 4.0 © 002350 0.25

Original plants... 0.952 29.8 18 5.5 0.003232 0.35

(I) 1¢.c. baryta solution =0,0031245 gram nitrogen,

Can Old Leaves of Plants Produce Asparagine

by Starvation ?
BY
T. Miyachi, Vogakushi.

It has been asserted that only young plants or growing parts
of plants, as germinating seeds or buds, can produce asparagine
by decomposition of proteids.

As it seemed to me of some interest to test the validity of
this assertion, I selected, on the 1st October, some old leaves of
Peonia albifiora, which by some brown spots clearly showed
incipient decay.

A portion was directly dried at 100° C, and served for the
determination of total nitrogen, protein nitrogen, and asparagine
nitrogen. On the same day, another portion was very loosely
placed in a glass vessel, containing a little water, and covered
by a glass plate. From time to time fresh air was introduced.

The microscopical examination, repeatedly carried on, re-
vealed a gradual disappearance of the starch granules from the
mesophyll (the epidermis cells were free from it at the beginning),
and also a gradual decrease of the active albumin, stored up in
the vacuoles, as the test, made with caffeine, indicated.”

On the 15th October, the leaves were dried, all brown
dead portions being first removed. Active albumin was still pre-
sent in the healthy parts, although in much smaller quantity than
at the beginning; but soluble passive albumen could not be
found.

The results were as follows :—

Fresh leaves. Starved leaves.
Total nitrogen. 1.364% 1.462%
Protein nitrogen. Teg 02 0.801

(1) The mean temperature was 17.68°C; the maximum 25.4°, and the minimum
16 b=

(2) Cf. this Bulletin vol. I]. No. 2 and 4.

(3) I crushed some of the leaves with some water, filtered and heated the filtrate
with a little nitric acid, but no precipitate was obtained.

(4) From this relative increase of nitrogen, it follows that 6.70 per cent of the origin-
al dry matter were consumed in the respiration process.

CAN OLD LEAVES OF PLANTS PRODUCE ASPARAGINE. 459

Fresh leaves. Starved leaves.
“Asparagine nitrogen. 0.037 0.200
Amido-nitrogen except asp. nitrogen. 0.015 0.455

In 100 parts of total nitrogen.

Fresh leaves. Starved leaves.
Protein nitrogen. 96.19 54.79
Asparagine nitrogen. 27a 14.09
Amido-nitrogen except asp. nitrogen. 1.10 Bitz

We observe, therefore, the proportion between asparagine
nitrogen in fresh leaves and that in starved leaves is 1: 5.2."

- The second experiment was made with tea leaves.

On the 19th October I collected old leaves of Thea chinensis
for the determination of total nitrogen, protein nitrogen, and as-
paragine nitrogen. At the same time numerous small branches
(about 10 cm. long), taken from the same individual plant, were
placed in a large vessel containing some fresh water, which was
placed in the dark for 24 days™ (Oct. 19th to Nov. 12th). All
the young leaves had been first removed. While the branches
were kept in darkness, the water in the vessel was renewed
several times.

Microscopical examination repeatedly made, revealed, as
usual, the gradual disappearance of the starch granules and the
decrease of active albumin, which was present in large quantity
at the beginning. Soluble fass¢ve albumin was not present at
all, as the test of the cold aqueous extract with nitric acid showed.

Since some leaves commenced to show brown spots, on the
12th November and further exposure was likely to end soon in the
death of all the leaves, I dried the still healthy ones, and sub-
jected them to the same examination as the control leaves at the
beginning.

(Bull. Coll. of Agr. II. No. 2) 2.222% total nitrogen, and 2.648% after 25 days’ starvation
by being kept at 25°—30°C in darkness in a covered glass vessel containing some water,
hence the loss by respiration amcunts to 16.09% in 25 days which when calculated to 15
days become 9.65%. The intensity of respiration in these young leaves was therefore
about one and half times as much as in the o/d leaves. The asparagine nitrogen was
there at the beginning 9.86% of the total nitrogen ; after 25 days, however, it became
45-73%- ‘The proportion between asparagine nitrogen in fresh leaves and in starved
leaves was there 1: 4 6.

(2) ‘Ihe mean temperature was 13.31°C, the maximum 23.7.C. and the minimum
3.2°C,

460 MIYACHI ;

The result was as follows:
In 100 parts of dry matter ;

Fresh leaves. Starved leaves.
Total nitrogen. 3.475% 3.987 %™
Protein nitrogen. 2.850 ZaG iar
Asparagine nitrogen. 0.201 0.867
Theine nitrogen. 0.283 0.358
Amido-nitrogen except asp. nitrogen. 0.141 0.251

In 100 parts of total nitrogen.

Fresh leaves. Starved leaves.
Protein nitrogen. 82.01 62.98
Asparagine nitrogen. 5.78 20.75
Theine nitrogen. 8.14 8.98
Amido-nitrogen except asp. nitrogen. 4.07 6.29

We observe, therefore, the proportion between asparagine
nitrogen in fresh leaves and that in starved leaves to be 1 : 3.7.

My investigation, therefore, places it beyond doubt, that
even old leaves can produce asparagine from protetds.

Some additional remarks may be made in regard to the be-
haviour of theine in the metabolism of plant cells.

In order to decide whether theine could be utilized as a source
of nitrogen and carbon for the formation of proteids, I prepared
50° of a solution containing 0.5% theine, 0.19§ dihydropotassium
phosphate and 0.019§ magnesium sulphate, sterilised it, and in-
fected it with spores of Aspergillus orizae. After standing from
the 8th May to the 18th June, only a very small amount of my-
celium covered with some spores, had developed.

In a second case the solution was infected with Penzcecliium
glaucum ; but no development was here observed.

Now, if theine proves to be sucha poor nutrient for lower
fungi, which possess great chemical energy, it may safely be in-
ferred that it will also fail to be a nutrient for tea-leaves. Indeed
its quantity is zzcrcased by the decomposition of proteids, as not

(1) From this relative increase of nitrogen it follows that 12.84 percent of the dry
matter was consumed in the respiration process.

(2) The high percentage of fatty matter present in tea leaves is evidently the reason
why the decomposition of proteid and consequently the formation of asparagine was less
energetic than in /7aeoz/a leaves.

(3) On shaking the solution with chloroform and evaporating, a considerable quan-
tity of crystallized theine was egain obtained, easily identified by the principal reactions.

CAN OLD LEAVES OF PLANTS PRODUCE ASPARAGINE. 461

only the above analysis but also the following placed beyond
doubt.

Old leaves of Thea chinensis, were analysed again both in
the fresh and in the starved condition. Small branches were
kept in water from the 23rd May to the 8th June at 19—23° C. in
a covered vessel in darkness.

The analytical results were as follows :—

Fresh leaves. Starved leaves.
Total nitrogen. 2.575% 3.187 %
Protein nitrogen. 2.149% 2070,
Theine nitrogen. 0.288 9% 0.438%
Amido-nitrogen. 0.139% 0.570%

The relative increase of total nitrogen from 2.575% to 3.187%
indicated that 19.21% carbohydrate, fat, and perhaps some carbon
and hydrogen from the decomposed proteids, had been consumed
by respiration.

A correct judgement can, of course, be obtained only when
the amount of theine-nitrogen in both cases is compared with that
of total nitrogen, which we may assume as a constant quantity in
both cases ; thus we obtain for 100 parts of total nitrogen 11.15 of
theine-nitrogen in the fresh leaves, and 13.74 in the starved leaves.
Assuming that no loss of carbon and hydrogen by respiration had
taken place, we obtain for the same amount of leaf-weight, i.e.,
for 0.996 gram. of theine in the original leaf, 1.227 grams of theine
in the starved leaf, showing that xo consumption of thetne had
taken place by starvation ; on the contrary an tncrease ts evident.

This result, compared with the observation on fungi above
mentioned, makes it highly probable that theine cannot be con-
sidered as a valuable nutrient.” It remains now to be seen,
whether zz presence of much carbhydrate and absence of any other
suttable source of uttrogen, theine (caffeine) can be used as a
source of nitrogen for building up proteids in tea leaves.

Analytical Data.

In the following analyses, the total nitrogen was determined
by AKjeldahl’s method, the protein nitrogen by that of Stutzer,
and the asparagine by that of Sachsse, while theine by that of

Mulder.

(1) This inference agrees with the observations of Errera, Maistriau, and Clau-

triau on the behaviour of alkaloids during germination (Bull. Soc. Belge de Microscopie,
1894).

462 MIYACHI;

Il. Peonia albifiora.

Total nitrogen in fresh leaves.

a). 1.8416 g. of dry matter contained 0.02512 g. of nitrogen
(=13.4°°™ baryta water) equivalent to 1.364%. b). 1.8416 ¢. of
dry matter contained 0.02512 g. of nitrogen (=13.4°° baryta
water) equivalent to 1.364% ; average 1.364%.

Protein nitrogen in fresh leaves.

a). 1.8416 g. of dry matter contained 0.02407 g. of nitrogen
(=12.84°-* baryta water)=1.3079%. b). 1.8416 ¢. of dry matter
contained 0.02426 g. of nitrogen (= 12.94°°™ baryta water)
= 1.397%, average 1.3120

Asparagine nitrogen in fresh leaves.

a). 4.604 g. of dry matter contained 0.000937 x 2“ =0.001874 ¢.
of nitrogen(=0.5 x2=1.°“ baryta water)=0.0407%. b). 4.604 g.
of dry matter contained 0.007490 x 2=0.0014¢8 g. of nitrogen
(=0.4 xX 2=0.8"° baryta water) =0.0325%. Average 0.0366%.

Total nitrogen in starved leaves.

a). 1.8584 ¢. of dry matter contained 0.02719 g. of nitrogen
(=14.5°° baryta waterJ=1.462%. b). 1.8584 ¢. of dry matter
contained 0.02719 g. (=14.5°° baryta water)=1.462% ; average.
1.462%.

Protein nitrogen in starved leaves.

a). 1.8584 ¢. of dry matter contained 0.015074 g. of nitrogen
(=8.04""" baryta water)=0.811%. b). 1.8584 g. of dry matter
contained 0.014700 g. (=7.84°°" baryta water)=0.790; average
0.801%.

Asparagine nitrogen in starved leaves.

a). 4.646 g. of dry matter contained 0.004875 x 2=0.009750 g.
of nitrogen (=2.6 xX 2=5.2°° baryta water)=0.209%. b). 4.646 ¢.
of dry matter contained 0.004687 x 2=0.009374¢. (=2.5x2=5° ©
baryta water)=0.2039% ; average 0.200%.

Il. hea chinensis (a).

Total nitrogen in fresh leaves. (Nov. 1895).

(1) 10% ¢m. of the standard sulphuric acid contained 0.237548 g. of sulphuric acid and
corresponded to 0.067873 g. of nitrogen; 10.¢-°™. of the sulphuric acid corresponded to
36.2¢- cm. of baryta water, 1¢-¢m. baryta water =0.001875 g. of nitrogen.

(2) A molecule of asparagine contains two atoms of nitrogen and as by boiling it
with hydrochloric acid only one atom is split off as ammonia, the result must be doubted.

CAN OLD LEAVES OF PLANTS PRODUCE ASPARAGINE. 463

a), 1.8182 g. of dry matter contained 0.063372 g. of nitrogen
(=33.8"° baryta water)=3.485%. b). 1.8182 g. of dry matter
contained 0.062997 g. (= 33.6" baryta water)= 3.465% ; average
3-475.6-

Protein nitrogen in fresh leaves.

a). 1.8182 g. of dry matter contained 0.05182 ¢. of nitrogen
(=97,64°* baryta, water)=2:6507¢. b). 1.8182 ce of dry matter
contained 0.05182 ¢. (=27.64°° baryta water)=2.850% ; average
2.850%.

Asparagine nitrogen in fresh leaves.

a). 9.0910 g. of dry matter contained 0.0091670 x 2=0.018334¢.
Of nitrogen, (=4.9 X 2 = 6.Saammaparyta water) = 0.202%: b):
9.0910 g. of dry matter contained 0.009000 x 2=0,018000 g. (=4.8
x 2=9.6°° baryta water)=0.199% ; average 0.201%.

Theine nitrogen in fresh leaves.

a). 4.5455 g. of dry matter yielded 0.046 g. theine(=0.013278g.
theine-nitrogen) =0.292% nitrogen. b). 4.5455 g. of dry matter
yielded 0.043 g. theine (=0.01241 ¢. theine-nitrogen) =0.273%
nitrogen ; average 0.283% nitrogen.

Total nitrogen in starved leaves.

a). 1.81730 g. of dry matter contained 0.072586 g. nitrogen
(=387"° baryta water) =3.992/p. b). 1.81730 g: of dry matter
contained 0.072371 g. (=38.6°°™: baryta water) = 3.983°/) nitrogen.

Protein nitrogen in starved leaves.

a). 1.81730 g. of dry matter contained 0.045823 g. nitrogen
(=24.44°° baryta water)=2.521%. b). 1.81730 g. of dry matter
contamed: 0.045448 9. (= 2ameaee baryta water) —2:5or °/,;
average 2.511 °/.

Asparagine nitrogen in starved leaves.

a). 4.54325 g. of dry matter contained 0.00294 x 2= 0.00788 "/y
mitogen) (= 10,5%2=21. ciemparyta water) 0.8666°/). bb).
4.54325 g. of dry matter contained 0.00394 x 2=0.00788 g.= (10.5
X2=21. c.cm. baryta water) = 0.8666 %; average 0.8666 %
nitrogen.

Theine nitrogen in starved leaves.

a). 4.54325 g. of dry matter yielded 0.058 g. theine
(=0.016742 g. theine nitrogen.) = 0.3689 nitrogen. b). 4 54325 ¢.
of dry matter yielded 0.055 g. theine (=0.015777 g. theine nitro-
gen) = 0 348 % nitrogen average 0.358 % of theine nitrogen.

464 MIYACHI.

Il. Lhea chinensis (b)'”

Total nitrogen in fresh leaves.

a). 0.9435 g. of dry matter contained 0.0242983 g. of nitrogen
(=14.2 c.c.. baryta water®)=2.575°/. b): 0:6435 c.noiednm
matter contained 0.0242983 ¢. (=14.2 c.c. baryta water)
= 2.575 "/o; average 2.575 Jp nitrogen.

Protein nitrogen in fresh leaves.

a). 0.9435 g. of dry matter contained 0.0203627 g. of nitrogen
(=11.9 c. c. baryta watenJ=2.158°/,. b). 0.9435 oof dryamat
ter contained 0.020191570 g. (=11.8c.c. baryta water) =2.140"/o.
average 2.149°/), of protein nitrogen.

Theine nitrogen in fresh leaves.

a). 4.7175 g. of diy matter yielded” 0045 2.) iiheme
(=0.0130412 g. theine nitrogen (=0.276°/, nitrogen. b). 4.7175 g.
of dry matter yielded 0.049 g. theine (=0.0141958 g. nitrogen)
=0.301°/); average 0.288 "/) theine nitrogen.

Total nitrogen in starved leaves.

a.) 0.8951 g. of dry matter contained 0.0285762 g. of nitrogen
(=16. c.c. baryta water)=3.192 9%. b). 0.8951 g. of dry mat-
ter contained 0.0284906 g. (=16.65 c.c. baryta water) =3.183 % ;
average 3.187 °/).

Protein nitrogen in starved leaves.

a). 0.8951 g. of dry matter contained 0.0194216 g. of nitrogen

= 11.35 ¢.c. baryta® water)=2.169°/, nitrogen.) ~~ b)y7e@:8o5 mes
of dry matter contained 0.0195927 g. (=11.45 c.c. baryta water)
=2.189 °/) ; average 2.179 "Jp.

Theine nitrogen in starved leaves.

a). 4.4755 g. of dry matter yielded 0.0635 g. theine
(=0.019195 g. theine nitrogen)= 0.428 °/) theine nitrogen. Db).
4.4755 g. of dry matter yielded 0.0695 g. theine (=0.0200618 g.
theine nitrogen) =0.448 °/, theine nitrogen ; average 0.438 °/o.

(1) These old leaves were collected in May 1896.

(2) 10 c.cm, of the standard sulphuric acid corresponded to 21.6 c. cm. of the stand-
ard baryta water and I.c. cm. of the baryta water corresponded to 0.00171115 g. of
nitrogen.

On the Relative Value of Asparagine as a Nutrient
for Phaenogams.
BY

T. Nakamura, Nogakushi.

It has repeatedly been shown that asparagine and other
amido-compounds applied to roots of plants are assimilated and
can serve as valuable sources of nitrogen. But quantitative
comparisons have not yet been carried out, which would no
doubt be of great value; above all it appeared desirable to
compare the relative value of asparagine as a source of nitrogen
with those of other closely related compounds. I made several
experiments to obtain some light in this direction.

In my first experiment I compared the action of asparagine
with ammonium succinate upon young barley plants, which were
measured from the base of the stem to the tip of the plumula at
different intervals. The solutions were prepared by mixing 2 %
solutions of asparagine and of ammonium succinate with equal
volumes of a saturated solution of gypsum.

After five days, the plants were placed for one day in an-
other solution containing mono-potassium phosphate and magne-
sium sulphate (0.1 99 of each).

Twenty four plants were placed in narrow beakers containing
200 c.c. of these solutions and kept at a temperature of 129°— 20°C.
After four days, the plants in ammonium succinate solution com-
menced to show a yellow colouration on the tips and this colour-
ation increased gradually so that on the tenth day all the leaves
had turned yellow, but they were still alive as the full turgor was
still preserved.

On the other hand, the plants in asparagine solution had
remained green up to this time. The great difference in growth
after nine days is seen from the following table :

(1) This separation of solutions is necessary to prevent bacterial growth, or to resist
it as much as possible.

466 NAKAMURA ; ON THE RELATIVE VALUE OF

Asparagine Ammonium succinate

5 cm. cm.

Average height, March 12th 20.27 19.92
Sy 34 pez tth 22.60 20.33
Difference Ne 0.41
Percentage of increase 11.49 2.06

In the next experiment, the solutions contained only one-
fifth as much asparagine and ammonium succinate as before, all
other conditions being the same. The result was after 14 days

as follows :
Asparagine Ammonium succinate

cm. cm.
Average height, March 23th 2a 21.06
6 - April 6th 28.13 24.08
Difference 6.82 3.02
Percentage of increase 32.00 14.34

In the third experiment, onion plants were treated with the
same solutions as in the first experiment, five plants being placed
in each of the solutions, after all the side leaves had been cut off.
Although in this case the central leaves in both solutions reached
the same height, there was, nevertheless, a great difference in
the namber and length of the newly developed branches.”

PLANTSeaN ASPARAGINE:

|

—
=
laws COT
TOIT rr,
—

(1) The leaves kept in ammonium succinate solution commenced to show yellow
colouration on the seventh day.

ASPARAGINE AS A NUTRIENT FOR PHAENOGAMS. 407

PLANTS IN AMMONIUM SUCCINATE.

—————
pn
un ane at pee
pi

Shaded branches are those newly developed.
Length of branches after 13 days.

Asparagine Ammonium succinate.)
cm, cm.
10.0 14.5
2220 | 10.0
9.0 23.5
5.0 (with a flower.) 7.0 (with a flower.)
27.0 17.0
GOI 45. py 8.0
19.0 23.0
10.0 103.5
TEOW st ort a
ZO
FO: sn oe
20.5
167.5

It thus appears that the total length of branches amounted to
50 °/, more with asparagine than with ammonium succinate.

(1) The difference in chemical structure is explained by the following formule :
CO-NH,

CO:O'NH,
CH, CH
CH-NH, ° bu,
COOH CcO:0'NH,
Asparagine

Ammonium succinate.

On the Relative Value of Asparagine as a nutrient
for Fungi,

BY

T. Nakamura, Nogakushi.

In order to arrive at a reliable judgement in regard to the
value of asparagine for fungi as a material for production of
proteids, it is necessary to offer it in conjunction with another
organic material which is free from nitrogen and is able not only
to supply a great deal of carbon which is necessary to form
proteids from asparagine, but which also would easily support
respiration and serve for the formation of cellulose. But such
very favourable nutrients as glycerol or glucose have to be
avoided, because these would somewhat obscure the relative
value of the nitrogenous compounds inasmuch as they yield a
very good result even with such simple nitrogenous compounds
as ammonia. I selected therefore as less favourable material
ethyl and methyl alcohol. The amount of nitrogenous and non-
nitrogenous compounds was such that the number of carbon atoms
was four times that of the number of nitrogen atoms. 500 c.c. of
-ach solution were taken and sterilized by boiling before the
alcohol was added.” The fungus used for these experiments
was <lspergillus orizae. In order to distribute as closely as
possible an equal number of spores upon the different flasks some
spores were suspended in 10 c.c. sterilized water and after well
shaking each time I c.c. was added to each solution. The flasks
remained in a room whose temperature (from 27th January to 15th
February) ranged between 5° and 15°C. The fungoid mass was
collected after 18 days on a weighed filter and after being
well washed, was dried at 100°C. The results were as fol-
lows :

(1) Each solution contained further 0.1 % monopotassium phosphate and 0,02 %
magnesium sulphate.

NAKAMURA ; ON ASPARAGINE.AS A NUTRIENT FOR FUNGI. 469

TABLE I.

Nourishing material in grams. for 500 c. c. Weight of fangus in grams.
Ammonium tartrate 6.33 + ethyl alcohol 3.06 0.015

# malate 5.00 + ,, 33 eS | 0.011

= succinate 5.00 + ,, 5 0,008

i lactate 7.13 + ,, FP, os 0.006

A acetate 5.13 + > 5 oF 0,000

. nitrate 2.66 4+ ,, 2. | 0,002
Asparagine 5.00 + ,, a a 0,026

“A 5.00 alone 0.016

Second experiment.

Here methyl alcohol was applied in addition to the nitro-
genous materials which were asparagine, glycocoll, urea, betain,
ammonium chloride, sodium nitrate, ammonium tartrate, am-
monium malate, and ammonium succinate. All the solutions
contained 1 °/, methyl alcohol, while the sources of nitrogen were
applied in such proportion that the number of nitrogen atoms to
those of carbon showed the ratio of 1: 8. This time the amount
of monopotassium phosphate was increased to 0.5 °/) and to all the
flasks ferrous sulphate,” sodium sulphate, and magnesium sulphate
(0.1 °/) of each) were added. All other conditions were the same
as before except that the volume of the solutions was only 200 c.c.
and the temperature somewhat higher, ranging from 9°— 19°C. The
flasks, after two weeks, exhibiteda considerable difference; in be-
tain, ammonium succinate and ammonium malate, no fungoid de-
velopment was yet noticed™; in ammonium tartrate only a moder-
ate quantity was observed, much however in asparagine and glyco-
coll. The flasks had been frequently shaken in order to prevent
the formation of spores on the surface which would have again
given rise to a great increase of the mycelium. Only during the
last 4 days shaking was dispensed with, allowing now spores to

(1) Certain fungi are only developed in presence of iron salts (A/o/isch) ; according
to Kau/in also presence of zinc salts will promote the development of mould fungi.

(2) They were infected again and left to stand for several months, whereupon a
moderate development was observed.

470 NAKAMURA.

be formed, the solutions containing asparagine and glycocoll
then showed more spore development than the other nutrients.

TABLE II.
Nutrient in gram for 200 c.c, solution Weight of fungoid mass in grams,
Ammonium tartrate 0.79 + methyl alcohol 2 g. 0.012
a chloride 0.42 + > e Z 0.025
Sodium nitrate 0.66 + 3 a os 0.015
Urea 0.25 + 5 3 Pr | 0,028
Glycocoll 0.79 + 3) # % 0.063
Asparagine O78 =e 4 + 0.073
= , Without methyl alcohol 0.047
Summary.

It is seen from these two experiments that for mycelium
fungi asparagine is a superior source of nitrogen, even far superior
to such a closely related compound as ammonium succinate.
Protein-production must then be much more easily possible from
the former than from the latter, z.e., the way from asparagine to
proteid is much shorter than that from ammonium succinate al-
though this is so closely related to asparagine, This conclusion
holds good also for phaeznogams as shown above.

On the Quantities of Nitrates Stored up in Plants
under Different Conditions.
BY

T. Ishizuka, Nogakushi.

The amount of nitrates stored up in plants varies not only in
different parts of the same plants, but it is subjected to great varia-
tion in the entire plants, which depends on the one hand upon the
relative amount of nitrates present in a soil and on the other upon
the state of development of plants, as, e.g., by rapid development
of plants all the nitrates, otherwise deposited in stem and root,
would be utilized for the formation of proteid.

I suspected, however, that, by gradual reduction, the amount
of nitrates would also decrease on keeping objects, especially
roots, for several months in a cool place.

A series of qualitative tests with diphenylamine and sulphuric
acid was first made to acsertain which objects are rich in nitrates;
this reaction failed, however, with the roots of Batatas edulis,
Nelumbo nucifera, Lilium tigrinum, Solanum tuberosum, Helian-
thus tuberosus, Capsicum longum, Eutrema Wasabi, Colocasia
antiquorum, and with the fruit of Cucurbita Pepo. The quanti-
tative determinations of nitrates were always made with fresh
objects, which were cut into small pieces and extracted with
water ona water-bath for half an hour; the residue obtained
by the evaporation of the filtrate was then extracted with alcohol
of 60°); this extract after evaporation to dryness served for
the determination of nitrates by the method of Tzeman and
Schulze.

The results are shown in the following table A.

(1) In some of the objects the qualitative test already revealed a decrease of nitrates
and in some cases an entire disappearance after several months, as in a variety of A//inm
Jéstulosum, On the other hand no decrease was observed with the root of Zappa major,
kept from gth October to 4th Nov., but here the examination showed at once tha! the
cells of the roots had died oft.

472 ISHIZUKA ;

Nitric anhydride in

Objects. Date of analysis. 100 parts of dry
matter,
Sept. 21. 0.13
Solanum Melongena.........0.0.0+
. Sept. 30. O.I1
2
3
a :
ei Oct. 24. 0.78
BCHUNCOSA) CHAS CT. Qian ne
Nov. 28. 0.52
Oct. I9. O.II
DD AUCUSN CAT OL O meanecen = iseeseee
Oct. 31. 0.06
Oct. 3. 1.35
| Altium fistulosumt..... cece
g Nov. 9. 0.17
2
3 Nov. 22. ay.
3 v 3:25
2 RaAPhANUS SAUVUS 0.0.0. cceceneen Febr. 24. : 2.85
Apr. 7. 2.50
Nov. 22. 1.16
Brassica CAMPECSUTIS.. 0.0 .ccceceeee
Febr. 29. 0.90
Nov. 9. 1.00
Brassica oleracea (sprouts)......
. Noy. 31. 0.95
zg
2
a Sept. 24. 4.14
&¢ Raphanus sativus . ...............
2 Oct. 21. 2.44
>
3
a
Sept. 24. 2.81
Brassica campestris ...............
Oct. 21. 2.12

From thts table it becomes quite evident that the amount of
uttrates in the different cases decreases more or less on keeping
the objects.

I directed my attention further to the amount of nitrates
found in the same plants at different seasons, especially in such
plants, as serve generally as food, viz., the root of Raphanus
sativus and Brassica campestris, commencing the determination

ON THE QUANTITIES OF NITRATES STORED IN PLANTS. 473

in October and taking samples from the field from time to time.
The differences here found were not so large as tn the other case,
where the objects were kept tn a state of rest after they had been
harvested.

Nitric anhydride in
Object. Root of Dates of collection. 100 parts of dry
matter.
ROP CHUSES QUUULS wecennasetecet ase 15 Oct 4.80
+ ms’ ch uehhiswaieseto ne stneles Di 4.13
53 ast Haisas wiemiinaceccseswe cael 28y 5, 4.77
6 Soe eceenecans desea lssieasias 4 Nov. 5.16
a pp MiessacsUinenacevaasapiter 1. 5 4.06
5 + eae esa eee ee as ae 3.26 (1)
os FR OSCR EE ea EE OS eaRnOSe 9 Dec. 2.50
3 SO ore ore ee 20 ,, 3 34
” fy dadadtinsodangaboancanptc 27 Jan. 2.66
ay Friis “WaROCLORedOCOnCEaE a ticad 12 Feb. 3.50
i. OS EERO RAE CEE RES 10 March. A 2.21
or RA ean derkas cotseisate neuer 29 Apr. 3.95 (2)
a9 Pee Cheroccbocoseeuaa occ: 22 May. 3.70
Object. Root of Dates of collection. Nitric anhydride in 100
parts of dry matter.
WS ROSSUCONCOIUPESTL US) eee aisnatnanasaetr 21 Oct. 2.27
rr aye: Cckrenmestenscanratereners 25) ,, 2.45
re Pe a ea seclle an nens ease 4 Nov. 2.36
3 i aa eee Reece: 1S: ,, 2.44
‘6 %) _-eBorebescosacppsvonte 9 Dec. 0.67
“ Perle caienicine deileacuicchien 20m, 1.06
i Ao, | hatcisalelote'sorersiarcie sietorare 27 Jan. 1,09
a7 Pe archer brace npannon 12 Feb. 1.21

(1) Frost had set in.
(2) This determination was made with a young plant.

474 ISHIZUKA3 ON NITRATES STORED IN PLANTS.

I paid also some attention to the products of transformation
of the nitrates. Ifall conditions are favourable, then of course
the nitrates are assimilated in the building up of proteids; if,
however, all conditions for this process are not fulfilled other pro-
ducts might be formed and I suspected above all a gradual trans-
formation into asparagine in such cases. To compare the
amounts of nitrates and asparagine in the roots of Raphanus,
Brassica and Daucus three determinations were made: first just
after they had been harvested, then atter being kept for sixty
days in the dark in moist saw dust at the ordinary temperature,
and again forty days later.

eee Risies of In dry matter.
‘J : examination. Percentages of Percentages of
nitric anhydride. asparagine.
22 Nov. 3-25 4.35
Raphanus Satvus.........0.00.000- 24 Feb. 2.85 5.66
7 Apr. 2.56 6.18
22 Nov. 0.048 5-47
Daucus Carota .......: Reh d 7 24 Feb. 2 6.44
7 Apr. 0.046 7.41
: 22 Noy. 1,16 6.20
Brassica CAMPESUVIS occ ccccee
24 Feb. 0.90 10.35

We here indeed observe a gradual decrease of nitrates
and increase of asparagine, but the latter increased so much
more that we must assume the larger portion of the asparagine
was produced etther by decomposition of protetds or by the trans-
Formation of other nitrogenous organic compounds.

On the Significance of the Nitrates Contained in
Plants for Animals and Men,
BY

T. Ishizuka, NMogakushi.

As the presence of nitrates in the food is of more influence
upon the well-being of animals and men than is often supposed, it
is well to discuss such circumstances as relate to the amount of
stored up nitrates in plants ; these are principally:

a). Intensity of nitrification in the soil.

b). The amount of rain removing the nitrates from the soil.

The amount of nitrates formed in the soil depends, of course,
above all upon the amount of ammonia present; in the second
place upon the condition of the soil.

Deherain noticed a great influence of the kind of soil upon
the intensity of nitrification: a porous soil formed more than
double as much of nitrate as a less porous one; a clayey soil
would admit less aération, therefore would be also less favourable
for nitrification (Ann. Agronom. 21, 353, 1895).

In the third piace the development of the microbes of nitri-
fication (Nitrosomonas and Wtromonas) depend not only upon
climate, and mechanical condition of the soil, but also upon the
presence of certain chemical compounds; thus it was found by
Dumont that the increase of potassium salts in a soil and the
simultaneous presence of humus and calcium carbonate favour
the development of the nitrifying microbes, and therefore the
production of nitrites from ammonia which very soon pass into
nitrates in the soil.

Furthermore, the amount of rain must have a great influence
upon the amount of nitrates, as, these are not absorbed by the
soil. Heavy rains combined with thorough drainage of the field
therefore deprive the soil well nigh completely of the nitrates
present. On the other hand, frequent small showers will merely
promote the energy of nitrification, and a larger amount of
nitrates will accumulate, because of the rain water in this case
not dratning away, but simply evaporating again from the soil.

476 ISHIZUKA; ON THE SIGNIFICANCE OF THE NITRATES

Moreover, shallow soils are more easily deprived of the
nitrates by the same amount of rain than deep soils ; and finally
a great influence will be exerted by the temperature, as in sum-
mer the nitrification is more intense than in winter. Thus we
observe a /ocal and temporal influence upon the amount of nitrates
present in the soil, and the great differences Berthelot” observed
in regard to the quantity of nitrates in plants thus finds a simple
explanation.

Countries with regular and copious summer rains will show
also less nitrates in the plants than countries with rather dry
summers, while again in desert-soils nitrates are not found at all,
because of no bacillus being able to thrive in the absence of
water.

The noxious qualities of vegetable food, rich in nitrates, for
animals have been recognized by Lawes and Gilbert, the illu-
strious investigators of Rothamsted, England. I quote from the
lecture of Henry Gilbert delivered in America in Nov. 1893 the

following passage :

“Then, again, as generally more or less of the
nitrogen in root will exist as nitrate, it will so far not
only have no food value, but it may be posttively injurt-
ous. It may be added that, other things being equal,
the higher the percentage of nitrogen in roots the lower,
as arule, will be the proportion of it as albuminoids,
and the higher that as amides and as nitrate, etc.
Further in direct experiments at Rothamsted with sheep
feeding on roots alone, it was found that while the
animals even gained in weight on 72fe roots, /ow in
nitrogen, they actually /os¢ on roots that were /ess ripe,
high tn nitrogen, and doubtless containing a larger pro-
portion of their nitrogen as non-albuminoid compounds.”

It is true that mztrates by themselves have not a very noxious
effect on animals as it requires about 2.5 g. of potassium nitrate for
1 kilo of body-weight of an animal to bring on death, but there

(1) Berthelot found that the amount of nitrates may vary from o to 15 % in potato,
from 0 to 2.8 % o in wheat, from 0 to 15 % in Amarantus. (Chem Centralbl. 1884,
639).

The amount in turnips and beets was found to vary between 0.5 —3.5 % of the dry
matter (Z¢ermayer, Physiologische Chemie der Pflanzen). j

CONTAINED IN PLANTS FOR ANIMALS AND MEN. 477

exist numerous kinds of bacteria which can reduce the nitrates to
the poisonous zz¢rztes while other kinds of bacteria again reduce
them directly to the less noxious ammonia.”

The dangerous character of nitrites is clearly elucidated
by the observation of Az¢kinson, that 0.2 g. sodium nitrite pro-
duces heavy intoxication in men.” Guinea-pigs are killed by
0.5 g. of sodium nitrite no matter whether administered through
the stomach or subcutaneously, under the phenomena of paralys-
is and kyanosis.

Of those bacteria which produce nitrites from nitrates, the
bacillus of Cholera asiatica acts most energetically, and Emmerich
and 7Zswbot have therefore propounded the theory that the
symptoms in cholera disease are due to the xztrztes formed by
thts bacillus in the intestines of men from the nitrates in food.
Indeed, this theory explains thus far alone the temporal, local
and zxdivtdual disposition for cholera.” Pettenkofer pointed out
that cholera in temperate zones makes its appearance as a great
epidemic always late in summer or at the beginning of autumn,
further that certain cities are never visited by this epidemic, and
that drinking water has no influence upon the spread of cholera,™
and inferred that there must exist a second principle besides the

cholera bacillus to make the genuine cholera possible.

(1) In this regard an observation made by Azchder is of great interest ; he discovered
nitrites in the urine in a case of acute affection (catarrh) of stomach and intestines, and
elucidated farther that the action of a coccus upon the nitrates of the food had given rise
to the production of the poisonous nitrites. (Fortschritte. d. Med. 13. 478).

(2) Certain animals as rabbits require more to affect them seriously ; perhaps there
exists cogditions in them by which the nitrous acid is prevented from being set free so
easily.

(3) Miinch. Med. Wochenschr, 1893. That theory was attacked by A7emperer and
Pfeiffer, \yat not disproved. By means of the reaction of Gries, nitrous acid would no
doubt be discovered more often than formerly in the feces of cholera patients.

For the production of the cholera-red reaction indol is necessary, but as Govini
(1893) has shown this is not formed in presence of much sugar and is absent sometimes
in cholera-feces.

(4) Semerad came to the same conclusion as Pettenkofer, that besides meteorological
conditions the soil has also great influence, as his investigations on the cholera-epidemic
in Fungbunz/au left no doubt on this point.

(5) fhe nitrates are present, if at all, in too small quantities to be taken into
account ; thus, the sum of nitrous and nitric acid in 10,000 parts of drink water of Tokyo
(1885) was found to be:

Jan. Feb. March April May June July Aug. Sept. Oct. Nov. Dec.
0.002 0.004 0.158 0.173 0.211 0.128 0,156 0.163 0.162 0.152 0.110 0,009

478. ISHIZUKA; ON THE SIGNIFICANCE OF THE NITRATES

He has also called attention to his observation, that great
cholera epidemics appear in those years in which the rain fall
of summer remains far below the average, which fact would
appear very strange, if the cholera bacillus alone were the
cause, as abundance of moisture would be most favourable to
the development of that bacillus.“ This fact as well as the
gradual decrease of the epidemic towards winter,” and further its
breaking out at the time when the new vegetables are harvested,
is in best accordance with the theory of Emmerich and Tsuboi,
for which my own investigations above described bring further
support. Finally I must point out that the law, discovered by
Pettenkofer in regard to the influence of rain, is also confirmed
by the phenomena observed in Japan: I compared the intensity
of the four cholera epidemics during the last 13 years with the
amount of rain-fall from May to October, and from this it becomes
evident that in those years in which the rain fall was considerably
below the average, the epidemic was more serious than in those
years in which the rainfall was more copious, as the following
tables show :

(1) I do not assert here that the nitrites produced are the exc/esive poison in cholera.
Indeed Ziipfe and Scholl have discovered in cholera cultures also poisonous proteids,
There exist cases of a milder form of cholera in which the formation of nitrites is not the
leading factor, as, e.g., also the cholera produced in Guinea-pigs by subcutaneous injec-
tions of comma bacilli. Cf. also the interesting article of //iippe, Ler). Klin. Wochenschr.
1894. No. 17.

(2) Montefusco tries to explain this by the decreasing virulence in great cold, but
this is not satisfactory, as cholera stops even in warm winters.

CONTAINED IN PLANTS FOR ANIMALS AND MEN. 479

NAGASAKI. POPULATION 71,600.

AMOUN? OF RAIN FALL MM. Absolute
number of

deaths from
June July August | Sept. E cholera.

92.1 aigtons 167.3 209.4
192.7 | 2314 | 358.2 | 309.9
985.4 | 149.6 | 159.2 | 117.4
369.1 135.1 192.0 253.5 (1)
324.0 | 154.8 192.8 322.7,
259-7 191.1 97-1 48.5
457.0 798.4 78.6 128.6

393.0 206.0 214.0 214.8

298.9 255.3 203.3 207.6
338.9 52.0 250.6

319.5 35.0 | 476.8
43-7 110.4 25.3
337.0 216.0 27.9

347.0 288.3 190.2

NAGASAKI.

TFMPERATURE (AVERAGE). °CELS.

May June July | August Sept. Oct.
1883 17.7 21.6 25.7 27.0 22.7 Sigs. |

4 sai Ss ) giartomat 25.6 25.6 24.1 168

5 13.2 | 21.7 25.1 27.4 24.1 18.7

6 18.2 ee 26.0 Zar 23.3 19.0

7 17.6 21.1 26.1 26.6 23.6 19.1

8 eae | | .. 262 27.8 23.4 Wey)
9 18.1 23.2 26.0 26.9 22.5 "om =
go 18.7 22.5 26.7 26.5 Cheon | a

I 18.3 21.8 25.5 26.6 25.2 19.2

2 Raney > a2 — one 24.4 17.6

3 18.5 21.6 27.9 | 26.8 25.4

4 17.8 24.5 27.8 29.1 24.3 18.5

5 Fame | open 24.1 27.0 24.3 17.6
[Mean i gasess Lemuonas 26.1 27.1 23.5 aaa ed

(1) Epidemic most violent.

480 ISHIZUKA; ON THE SIGNIFICANCE OF THE NITRATES

HIROSHIMA. POPULATION 94,300.

AMOUNT OF RAIN FALL MM. Absolute
desta tea
May June July August | Sept. Oct. cholera.
~ 3883.—O«|:S 51.6 10.7 204.6 115.9 116.4 50.2 ~~ e
4 163.0 198.3 328.6 126.2 167.6 59.2
5 251.6 563.7 111.9 57-3 1298 85.2 13
6 249.9 131.4 135.1 192.0)} 253.5 IQI.I 5229
7 162.4 224.2 139.3 38.6 153.6 145.8
8 86.8 181.1 255-4 50.4 176.9 51.8
9 107.1 329.3 610.7 62.8 139.8 126.2
90 | 185.3 | 245,0 | 1609 | 809 | 165.5] 99.0 | 1325
I 178.2 252.6 210.8 79.3 163.2 io8- |) an
2 | 2279 | 2yseliees | 344 | Grea [Jer
3 igorom 1420 6.3 214.0 166.7 308.1
4 78.0 "T30g0n 153.5 66.3 328.4 ae
5 242.9 244.7 127.3 50.6 (4) 3049
Mean 165.9 225.9 203.4 95-7 163.2 112.7

HIROSHIMA.

TEMPERATURE (AVERAGE). °CELS.

June July August Sept.

26.0 27.0 23.0

25.0 25.1 23.6

24.3 23.5

26.1 - 23.1

22.6

21.8

21.5

24.0

24.4

(1) Epidemic most violent.

CONTAINED IN PLANTS FOR ANIMALS AND MEN. 481

OSAKA-FU. POPULATION 490,000.

Absolute
number of
deaths from
June July | August | Sept. Oct. cholera.

AMOUNT OF RAIN-FALL MM.

6.4 107.9 84.6 121.9 Wie

215.0 307.8 48.2 265.0 25.1

867.4 112.2 26.7 65.9 166.101)

144.9 8.6 24.0(2)| 187,1 138.0

211.1 733 98.6 97.2 221 4

119.4 145.8 86.1 120.1 92.8

165.8 231.3 193.8 156.5 163.0

232.4 118.1 50.7 129.3) 154.1
222.9 132.8 68 6 132.7 155.2

262.6 169.4 590 1688 143.1

112.4 14.7 1165 182.5 1850

97.1 95.1 81.7 109.3 84.0
279.2 208.1 87.9 147.7

Mean 228.2 131.1 78.9 144.9 133.2

OSAKA.

TEMPERATURE (AVERAGE), CCELS.

May June July August Sept. Oct.

16.5 21.3 26.2 26.7 22RT 17.6

16.8 21.0 25.2 25.4 23.4 15.5

16.7 21.9 25.1 27.3 23.9 17.9
17.4 21.8 26.7 27.6 23.7 18.0

16.3 21.2 25.7 Die 22.8 18.6

18.0 20.4 26.3 27.3 22.6 16.1

16.4 22.3 25.2 27.1 21.6 16.3
17.9 22.7 25.8 27.2 25.0 17.0
18.3 21.3 25.6 26.5 25 0 16.6

16.7 22.0 27.0 y/o 24.5 17.0

16.6 20.7 27.5 27-4 24.6 L7au
17.5 24.1 28.2 28.4 23.4 16.4
18.2 21.5 244 27.8 23.8 17.4

17.2 20.9 26.2 Pfc! 22.9 17.8

(1) Epidemic most violent.

482

AMOUNT OF RAIN-FALL MM.

TOKYO-FU. POPULATION 1,342,000.

ISHIZUKA ; ON THE SIGNIFICANCE OF THE NITRATES

Absolute
number of

deaths from
May June July August Sept. Oct. cholera.
1883 92.4 199.0 90.8 i 1.9 | 135.4 | 163.5
4 134.9 177.5 102.9 90.5 193.2 78.6
5 Wee 331.0 182 6 103.2 72.0 296.9 112
6 164.1 80.5 48.5 87.1 254.6 190.9 9879
7 147.4 | 216.2 g12 97.6 102.3 | 223.5
8 144.4 174.0 135.9 81.0 184.5 Patt toy
9 195.3 68.5 259.9 96.2 187.3 109.3
90 141.5 188.8 120.1 106.0 214.3()| _ 198.0 3307
I 347-5 180.3 130.1 105.3 212.5 IQI.7
2 267.9 285.9 109.1 20.9 288.9 136.6
3 145.3 95.5 54.9 95-3 90.8 ere a
4 84.3 | 575 | 616 | 1995 | 1449 | 192.7
5 177.6 299.8 129.3 83.6(1) 2500
sy flor) 109.0 101.8 166.4

TEMPERATURE (AVERAGE),

BOK YO:

June

July

August

°CELS.

Sept.

19.6

230

24.8

21.6

19.5

23-3

20.3

2351

23.8
25-5

20.8

25.0

26.5

203

23.6

18.6

24.5

25.3

21.4

22.1

23.3

21.1

25.6

2T0O

20.9

23.3

25.8

22.0

235

254

20.3

24.9

21.1

25-7

25-5
26.3

25.2

26.1

23.6

26.8

27.0

22.1

25-5

24.2

25.6

Epidemic most violent.

CONTAINED IN PLANTS FOR ANIMALS AND MEN. 483

The circumstance, that Japan has, as a rule, much rain in
summer, may account for the fact that the cholera-epidemics, so
frequent in Japan, rarely assume such frightful proportions as
are observed in certain cities in Europe, as Munich, Hamburg,
Marseilles, Naples or Palermo.

One of the violent epidemics in Japan was that of 1886 in
Osaka and just in that summer the rains in June, July, and
August were very far below the average, while they were
much less so in the case during the three other milder epidemics
of 1885, 1890 and 1895. The same rule holds good further for
the epidemics in Tokyo, while in Nagasaki and Hiroshima the
four epidemics reached but small dimensions, and here the
meteorological tables again show that the average amount of
rain in June, July, and August never sank so far below the
average as was thercase in the year 1886 in Osaka. We may
therefore conclude that Pettenkofer’s conclusions are confirmed
by the observations in Japan.

(1) As in spring and summer of 1896 numerous and heavy rains fell in Tokyo,
I predicted that a cholera-epidemic would not take place in the following autumn.
And indeed during the months of August and September the number of cholera cases
for every week were restricted to an average of three or four, gradually disappearing
ontirely in October. O, Loew.

On the Physiological Behaviour of Maleic

and Fumaric acids.
BY
T. Ishizuka, Nogakushi.

The stereo-isomeric fumaric and maleic acids,

HOOC—CH CH—COOH
I] and || show not only in chemical,
CH—COOH CH-—-COOH,

but also in physiological respects highly interesting differences.
Mould fungi can easily utilize fumaric acid as source of carbon for
building up their protoplasm, but not maleic acid.” Lew found
a similiar difference for bacteria; fumaric acid is an excellent
nutrient, while maleic acid an exceedingly poor one.;

Moreover, Pfeffer found that maleic acid exerts an attracting
influence upon spermatozoids of ferns, while fumaric acid shows no
such action.

Again, recent investigations show that maleic acid is more
poisonous for the higher animals than fumaric ; if per kilo body-
weight of a dog are injected 1.94 grams of maleic acid in the form
of the sodium salt into the veins it suffices to kill the dog, while
with fumaric acid it will not.

It appeared to me of some interest, to investigate, whether
analogous physiological differences exist in regard to chlorophyll-
bearing plants and the lower aquatic animals.

Preliminary trials showed that highly diluted solutions of the
neutral sodium salts of both acids did not exert any noxious in-
fluence upon plants; but it may be possible that maleic acid is
transformed by the vital activity into fumaric, before it can
exert a noxious action. The fact that there is never found in
the vegetable kingdom maleic, but fumaric acid is,” renders this
supposition indeed admissible. Therefore, the neutral sodium
salts of maleic and fumaric acids were applied in higher concen-
trations, and here indeed it became evident that maleic acid acts
more noxiously than fumaric acid.

(1) £. Buchner. Ber. D. Chem, Ges, 24, 1163.

(2) Central-Bl. f. Bact ; 12, 361.

(3) Hodera, Chem, Ztg., Dec. 1895.

(4) Fumaric acid was found in “umaria officinalis, Corydalis bulbosa, Glaucium
Zuteum, farther in different kinds of Agaricus and in Cetraria islandica.

PHYSIOLOGICAL BEHAVIOUR OF MALEIC & FUMARIC ACIDS. 485

I. Experiments with Leaves.

Young leaves were placed on the surface of 1 % solutions of

the neutral sodium salts of both acids and at the same time in
common water for control.

In fumaric
acid.

In maleic

: n water.
acid. I te

Leaves of

weet eee eterecce ” ”

11)

Experiments with Whole Plants.

Barley plants were placed in 2°/) solutions of the sodium
salts ; after 20 hours they were found to be killed by the sodium
maleate but were still quite normal in the fumarate, just as in the
control case with water.

Ill. Experiments with Branches.

Small young branches 10-15 cm. long bearing leaf-buds, or
flower-buds or both, were placed in 1°/) solutions of the neutral

sodium salts and at the same time in common water for control.

Branches of In maleic Tn fumaric Control.
acid. acid.
Oenanthe stolonifera ...... Killed after 4 days. | Killed after 10 days. Alive.
Acanthopanax spinosum.... : Py ay, 5 3 Or, 53 oe
Brassica campestris... 53 5 Sa es 0 ends os
Evonymus Thumbergianus iy i Sees 5 moe ”

Prunus pseudocerasus var.
Spontanea 12

buds,

bearing

Frunus mume bearing 5

flower buds.

Killed after 2 days,
no bud had open-
ed at all,

No buds opened,
after 3 days the
ends of the buds
turned brown and

died.

After 1 day 10 buds
had opened, and
on the 3rd day 2
more buds ;
the 4th day how-
ever they died.

on

After 1 day one bud
opened, but after
4 days all were
killed,

All buds opened,

and
alive.

remained

After 2 days all

the
opened.

buds

had

486 ISHIZUKA. ~
IV. Experiments with Seeds.

Seeds of barley and radish, six in number were soaked in
2 °/y solutions of the neutral sodium salts for 2 days, then placed
on moistened blotting paper under a bell-jar for germination.
After 10 days two barley grains had germinated of those that had
been treated with the maleate, three of those with the fumarate
and five of the control grains soaked in common water: of the
radish seeds all had germinated within 8 days, but the control

seeds already within 4 days.
V. Experiments with Algae.

Analogous experiments were made with filaments of Spzro-
gyra which were placed in 1°/, neutral solution of the sodium
salts of both acids: the microscopical examination after 4 hours
showed that about half the cells had been killed by the sodium
maleate, while only very few in the fumarate: after 18 hours all
cells in the former solution had been killed, their chlorophyll
bodies had lost their normal shape, and the cytoplasm had been
much contracted, while in the latter solution about half the cells

were still alive ; it took here 40 hours to kill them all.

VI. Experiments with Aquatic Animals.

The organisms principally observed were infusoria, rotatoria,
and copepoda.

All these remained alive in 1 p.m. solutions for several days;
but in 5 p. m. solutions they were killed in the sodium maleate after
1 hour 20 min., while in the fumarate after 8 hours : most copepoda
and rotatoria died in the former in 45 min., in the latter after
2 hours 3 min. We observe, therefore, in all the cases described
here, that szalete acid shows amore poisonous action than fumaric,
another interesting instance demonstrating the sensibility of the
protoplasm towards stereo-isomeric bodies. .

On the Physiological Action of
Amidosulphonic Acid.
ae

N. Maeno, Nogatushi.

Amidosulphonic acid has been subjected to extended chemic-
al investigations by Dr. Edward Divers, Professor in the Imperial
University of Japan, and it was he who proposed to test whether
this substance would prove just as good a source of nitrogen. for
plants as the ammonium salts or whether they would be noxious,

Dr. Loew then made some experiments which showed that,
while bacteria and mould fungi could utilise the sodium. and
calcium salts of this acid as soures of nitrogen, and algae are not
noxiously affected even by 1 per cent solutions of these two salts,
higher plants, on the contrary, are noxiously affected by them.
As this fact was so contrary to expectation, a larger series of ex-
periments appeared desirable, to decide whether such an action
would generally take place in different groups of phanerogams.

I therefore made experiments not only with entire plants and
young branches but also with isolated leaves and with the seeds
of various species. It is well known that ammonium salts in a
certain concentration act noxiously upon plants as well as upon
animals, and that this poisonous nature either decreases or in-
creases in intensity when we substitute one hydrogen atom with
other groups; thus, hydroxylamine and diamidogen are more
poisonous than ammonia, while on the other hand amido-acetic
acid or taurin are not poisonous :

NH, Weak poison.
NH,—NH, .

St s.
NH,—OH rong poisons
cae ae eae { Not poisonous.
NH,—CH,—CH,—SO,0H

Nenckt has found that carbamic acid exerts a poisonous effect
upon warm-blooded animals, which is different from the effects of
the equivalent amount of ammonia. This proves that hydrolysis
into ammonia and carbonic acid is not readily accomplished by

488 MAENO.

the living cells, and such may also be the case with amido-
sulphonic acid taken up by the plants.”

NH,COOH NH,SO,OH

Carbamic acid. Amido-sulphonic acid.

In ail my experiments with whole plants, young branches
and isolated leaves, I used a I p. m. or 0.5 p. m. amido-sulphonic
acid solution in the form of the calcium salt.” To these solutions
were further added 0.05 9% mono-potassium phosphate, 0.05 °/,
magnesium sulphate and 0.2°/, calcium sulphate. The control-
solution contained in place of amidosulphonate, an equal amount
of ammonium sulphate.

At the same time one or more of the plants were also placed
in distilled water.

We will call for the sake of abbreviation the plants in the
solution containing the amidosulphonate (A) ; those in ammonium
sulphate (B), and the plants in distilled water (C).

I. Experiments with Whole Plants.

Barley plants were taken carefully from the field on the 19th
February and after carefully washing the roots, they were placed
in the solutions mentioned. The result is seen from the following
table:

Length of plants at | Length of plants Water absorbed in
the beginning. after 9 days. 6 days.
A 24.5 cm. 24.5 cm, 46 c. c.
B | ZA‘OL ;; 25.0" 5, 105 5,
Cc 2 Our 23.0 ,, 120 ,,

The plant (A) appeared so much damaged by the withering of
the leaves on the gth day that complete death set in two days
later ; the chlorophyl of the younger leaves had turned yellow
before they succumbed. While no traces of new rootlets were to
be observed in the plant (A), the control plants which remained
healthy for a long time had developed them in great numbers.

(1) According to an observation of Dy. Divers to whom thanks are due for having
provided our laboratory with a large amount of amidosulphonic acid, only the am-
monium salt of this acid is easily hydrolysed in aqueous solution,

(2) In a few cases I also used the sodium salt with the same results.

PHYSIOLOGICAL ACTION OF AMIDOSULPHONIC ACID. 489

For the next experiment young plants of Brassica Rapa
14. cm. in height were selected. Although the amount of amido-
sulphonic acid here did not exceed 0.5 p.m: the plants (A) died
within 7 days, while the control plants B and C remained
healthy.

In the third experiment plants of Alium fistulosum about
30 cm. in length were placed in 0.5 p. m. solutions.

A noxious effect was here noticed after 14 days, the tips of
the leaves became yellow, lost their turgor, and withered. Com-
plete death had set in after 29 days with the plants A, while the
control plants B and C still exhibited a healthy appearance.

The plants A had grown during the first 7 days 9 cm. on
the average, but the plants B more than double that height.

For the fourth experiment young plants of Soya beans
were used. The seeds were soaked for several days in water and
then left to germinate on moistened filter paper. The shoots
were placed in 1 p.m. solutions. In this case not only the amido-
sulphonate but also the ammonium sulphate exhibited a decided-
ly noxious action.

Length of the shoots.
= Death set in:

At the beginning : After 11 days:
A 8 cm, 8 cm. In 11 days.
B 9 » 9 » Vim Hit 2
€ 8 1S) —

Il. Laperiments with Branches.

In the 1 p. m. solutions above mentioned were placed on the
3rd of March, small twigs of Prunus domesticus 12-15 cm.
length and bearing flower-buds; the buds of those twigs that
were kept in water opened on the next day. One day later, the
buds of the branches kept in ammonium sulphate followed and 4
days later, those of the branches in amidosulphonate (A). On

(1) Diamidogen in a dilution of 0.5 p.m. killed the plants in about the same time as
amidosulphonic acid.

490 MAENO ;

the 10th of March, however, the 4 blossoms (A) had dried up
while the still closed buds died in this state. The branches in
water, however, had 14 and those in ammonium sulphate 11
healthy blossoms. In other experiments, the solutions were ap-
plied in half the concentration as before but the result was essen-
tially similar ; after 8 days, all the 14 buds on the branches (A)
had withered and become brown, while the branches (B) and (C)
with all their opened buds remained healthy.

Ill. LAxperiments with Leaves.

Young leaves of Aesculus turbinata 15-17 cm. in length were
placed in 1 p. m. solutions.

After 4 days, a loss of turgor could be observed with the
plants A, and several days later black spots appeared, especially
along the veins, which spread gradually and extended over
all the leaves after 11 days. The leaves B and C, however,
retained their normal appearance. This experiment was repeated
with solutions of half the concentration with essentially the same
results. In the next experiment, leaves of Prunus cerasus
(5 pieces) were placed in 0.5°/, solutions. After 5 days, brown
spots developed on the leaves A, which after 8 days had spread
so much that the leaves might be considered as completely killed.
Soon afterwards, the solution in which these leaves were kept be-
came reddish from extracted organic matters.

The leaves B showed at this time only very small and few
brown spots, while leaves C showed only one such spot of very
small size.

IV. Experiments with Seeds.

Seeds of rice, barley, soya bean, and turnips, 20 of each,
were placed for 58 hours in 0.5°/) solution of calcium amido-
sulphonate (A); in ammonium sulphate (0.5 °/)) (B) and again in
pure water (C).

The seeds were then placed on moist filter paper, covered
with a bell jar, and the number of germinated seeds counted
every evening for twelve days. The result is given in the follow-
ing table :

PHYSIOLOGICAL ACTION OF AMIDOSULPHONIC ACID. 4091

OYaDeaN -o.-.0--2---

TRS gcone ooeeneeee B Onl 2a aers | 27 | 27) esa ou! x9) 19) |) 9)/ 19

G o| 6| 15-| 18} 18 | 18 | 18} 18 | 18 | 18] 18 | 18

These results demonstrate again the noxious effects of the
amidosulphonic acid, but how the differences in poisonous intensity
are to be accounted for, [am unable to say. Perhaps the calcium
salt of that acid penetrates more easily into the embryo of one
kind than into those of the other. Thus it might also be explained
why seeds of buckwheat and sunflowers were not damaged at all
by the same treatment as killed all the soya germs completely
(see table).

V. Experiments with Yeast.

It appeared to me of some interest to see whether the amido-
sulphonic acid would also show a noxious influence upon yeast.

For this purpose, I distributed 10 cc. of thick beer yeast in
distilled water and diluted the mixture to 100 cc. After well
shaking, I took 10 cc. of the mixture immediately after shaking
and added gocc. of a glucose solution containing 6.856 grams
pure glucose, 0.1 gram magnesium sulphate, 0.2 gram dihydro-
potassium phosphate, and 0.1 gram. sodium amidosulphonate, A.

In the control case instead of the last, ammonium sulphate
was used, B.

402 MAENO;

10 cc. of the diluted yeast applied corresponded to 0.0613
gram dry matter.

After 5 days’ fermentation, the mixtures were filtered off and
on the one hand, the dry matter of the yeast, on the other, the
amount of sugar still present were determined.

The yeast A weighted 0.165 gram, the yeast B 0.198 gram.
The volumetrical sugar determination still showed in the flask A
3.52 gram, in B, however, only 3.07 gram, of sugar.

1
| ix B
|
= = = —
Imcrease of yeast in ‘erams) 2. psseseeeeeeeeeereee | 0.1037 | 0.1367
Increase of yeast in percentages of dry yeast. | 169 %. 223 %.
Sugarifermented inigyarms) eee seeeeeeeeeee 3 3.3360 3.7860

Sugar fermented in percentages of dry sugar. | 48.80 %. 55:20 %.

We see, therefore, that the amidosulphonic acid does not
prevent the growth and fermentative power of the yeast, but it is
a less favourable source of nitrogen than ammonium sulphate,
which again is a surprising fact, as the chemically powerful yeast-
cells should in our opinion be capable of easily bringing on the
hydrolysis of the amidosulphonic acid.

VI. Experiments with Mammata.

I made in this regard but few experiments. Into a white
mouse was injected subcutaneously 0.5 cc. of a 1°/) solution of
sodium amidosulphonate. Since I could not observe any noxious
effects after 48 hours, I injected once more 1 cc. of the same
solution. Soon afterwards a considerable increase of the respira-
tory activity was noticed, but 2 days later the mouse was ina
normal state again.”

In another experiment, I soaked bread in a 1°/) solution of
sodium amidosulphonate and fed a mouse with it. This animal
became gradually very weak and somnolent and died after
76 hours. In this case perhaps such a large quantity of the

(1) This result agrees with those obtained by Prof. Takahashi in the Imperial
University in Tokyo, The observations of Prof. O. Zoew on lower aquatic animals,
which remained alive in 1 p. m. solution of calcium amidosulphonate, are also in accord-
ance with my results.

PHYSIOLOGICAL ACTION OF AMIDOSULPHONEI ACID. 493

amidosulphonate was introduced into the body that no safe con-
clusion as to its poisonous character can be inferred. Mencki had
observed that 0.6 gram of sodium carbamate per kilo of body
weight upon injection into the blood of a dog produces tetanical
convulsions and sometimes death, 0.3 gram per kilo will produce
somnolence” and catalepsy. As in the above first mentioned
case the relative amount of amidosulphonate was nearly the same,
it becomes evident that amidosulphonic acid is not so noxious
to animals as the related carbamic acid.

To summarise:

Amidosulphonic acid occupies an exceptional position among
the poisons: it is neither poisonous to higher or lower animals,
nor to fungi and algae, but it is poisonous to all kinds of phzno-
gams. Although no poison for fungi it is not so favourable a
source of nitrogen for them as ammonium salts.

(1) Carbamic acid continuously produced in the body, is rapidly transformed into
urea, nevertheless it may possibly exert some influence upon the causation of the normal
sleep. Cf. on this point also the interesting publication of Zeo Lrreva, Sur le Mecanisme
du Sommeil, Brixelles, 1895.

Investigations on the Mulberry Tree.
BY

N. Maeno, Nogakushi.

I. Lmprovements tu the Quality of Mulberry
Leaves by a Special Manure.

It is a fact that in certain provinces of Japan a better sill is
produced than in others, although the silk worm is sometimes of
the same variety. It may be surmised that the nature of the soil
exerts much influence on the quality of the leaves used as food
by the caterpillar. The relative amount of digestible and indi-
gestible material in mulberry leaves must naturally have a
certain bearing upon the well-being of the silkworms and hence
also upon the quality of the thread produced. This led me to in-
stitute some experiments with the intention ofattaining a decrease
in the amount of woody fibre and especially an increase in the
amount of proteid and fat. I supposed that manuring with lime,
calcium sulphate, and sodium nitrate would be especially well
adapted for the production of such a superior quality of leaves,
and thought it best to apply lime in the form of slaked lime, hop-
ing thereby also to destroy the mycelium of a very noxious
fungus,” found in our mulberry plantation at the College of Agri-
culture in Tokyo.

The soil consists here principally of volcanic ash mixed with
sand, and contains from 7-8 % humus. It is rather poor in lime
and sulphates and had received, one year before I commenced
my experiment, a moderate dose of night soil as manure for the
trees. I manured a mulberry tree about 1$ meter high with 500
grains lime, 400 grms. sodium nitrate and 200 grms. calcium
sulphate” in the beginning of March (A), while another tree was
manured with 500 grms. lime alone (B); a neighbouring tree re-
ceived no manure and served for comparison (C).

(1) Helicobasidium Mompa, studied by Tanaka, Journal of the College of Science,
Tokyo, Vol. IX.

(2) These materials were well mixed with the soil to a depth of about 30 cm. and to
an extent of one square meter around the tree. I wanted to have phosphoric acid,
potassa, and magnesia in the minimum, and supposed there was some left in the soil
from the year before.

INVESTIGATIONS ON THE MULBERRY TREE. 495

On the 20th May, a number of leaves were collected from
each tree for analysis ; each leaf measured on the average 9 cm. in
length and weighed on the average (of 50) in case (A) 0.320 gram ;
in (B) 0.304, in(C) 0.303 gram. On analysis, I obtained the follow-
ing results.

A B C

WW AEST 9,5 tiene, Fis seers 2 EE © + 80.82 80.85 80.04
Diyematter ~...2.. 20425» 19.18 19.15 19.66
Organic matter (in the dry substance)91.63 | 90.98 Ol 19
AAG) Se ae ees... a 8.37 9.02 18.81
PMDC i te ake stock ee: 13.10 13.68 18.11
At Tot esas sGi tS... 5.34 4.56 4.49
Motal carbohydrate. ...> 2: - - 23.14 22.92 23.44
Crude protein: 2... ....¢ eee - -28,56 23.31 23.25
(otal nitresen\....... 2 eee... 4.25 3.73 B52
Albuminoid nitrogen .....) eee 3.47 3.29 3.28
mMido=nitrosen .... . .<5 2 eee: - 0.78 0.44 0.44
Non-nitrogenous extract (except

Canbohydrate) &..<%.a. see +» 21.49 26.51 21.90

We observe here that by liming alone, the percentage of
woody fibre decreased from 18.10 % to 13.68, while the non-intro-
genous extract had increased from 21.90 to 26.50 % ; further by
liming and manuring with sodium nitrate and calcium sulphate;
not only had the amount of fibre deceased from 18.11 to 13.10 %
but the proteid had increased from 23.25 to 28.56 % and the fat
from 4 49 to 5.34 %.

It can not be denied that this special manuring produced
leaves of superior nourishing quality and it remains to compare
the effect of these different leaves upon the silk worm.

Il. Ox the Amount of Reserve Material in the Bark

of the Roots and Pranches of
the Mulberry Tree.

It appeared to me of some interest to determine the extent
to which the reserve material deposited in root and branches is
consumed in spring time.

On the 25th January roots about 1 cm. in diameter were col-
lected whose bark was separated for analysis (a). This was re-

(1) In September of the same year the tree B had gained in height one fourth and
the tree A about one third compared with the control tree C.

496 MAENO ;

peated with roots of the same tree and nearly of the same diameter
on the 27th April (8), when the leaves were developed. The
analysis shows that a decrease of proteids and non-nitrogenous
extract had taken place on the one hand, while on the other hand
an zverease of starch; fat and fibre had remained nearly constant
while lecithin disappeared almost completely.” In connection
with the decrease of proteids, an increase of amido-nitrogen and
especially of asparagine nitrogen is of interest.

Also the dark of the branches was subjected to analy sis at
two different periods.

The sample (a) was collected on the first of March before
the leaf buds opened and the sample (f) after the development
of the leaves on the 27th April.

Here we observe not only a decrease of proteids (with in-
crease of asparagine) but also a decrease of fat, lecithin, and total
carbohydrate.

The result is seen from the following table :

Root. Branch.
(a) (B) (a) (B)

Watt os jones cats a 67.01), 70.385 53:2%/nn ys -ooun
Dry.matter =. saa aes 2-00 29 62 40.80 26512
Organic matter = 27>... eeeo0-43 g6.78 94.10 94.02
ASH: a0 Sos ke oo eee 8.57 2.22. 5.90 5.98
Bibte\ 29). Ae eee B1.130 32420. 35-770 AG5Co
Bat> fl de:. sk. Bao. ees ee 6.700 6.300 7.290 6.700
Ibectthin . 20 et, ee 0.999 trace 1.076 trace
Fotalcarbohydrate-:2seeee 15.040 26.500 19.800 17.200
Sugars and dextrin 25 0eaem 6.600 10,500 ~13:300 9.070
Starch: 2.64.40. he02 eee 8.440 16.000 6.500 8.130
Crude protein \....255 See 9.450 FAST 2 Ossie 9.956
Total nitrogen 32 eee 1.512 1.150 1.650 1.593
Albuminoid nitrogen........ 0.898 0.394 1.294 1.040
Amido-nitrosen’..-.. 425 eee 0.094 0.100 0.005 0.003
Asparagine nitrogen........ 0.520 0.656 PAS 0.550
Non-nitrogenous extract (ex-

cept carbohydrate). 7 -.eee 33-111 27.592 26.827 19.644

(1) I may here draw attention to the recent observations of S/okéasa upon the
formation of lecithin in the leaves in daylight and its disappearance in darkness.

INVESTIGATIONS ON: THE MULBERRY TREE. 497

Analytical Data.

1. Determination of fibre.

Dry matter in grms. Fibre in grms. Percentages.
ROGt(@)i nc.c hws oe h. 2, 2IZRO 0.84g0 B13
Seat (DB) oom 2 det Ras ee eee 0.9250 32.42
Ba (O2)! = oo Oboe Die ane OOO) 0.0940 35 77
pee (Vf) Scions ete d cn ce OO 1.2605 46.50
Beate CA)icc. 0.263 ae 1.8505 0.2425 13.10
So (8) eRe ke Bae 1.8385 0.2515 13.68
WE) Barnet 2 eats ns ool ae 1.8270 0.3210 18.11
2. Determination of fat.
Dry matter in grms. Fat in grms. Percentages.
LGo 71/0) ee a ee 3.6360 0.2440 6.70
(/ SUS eee = 3.8048 0.2440 6.30
535.16]! \(0'0) eee ee ee 3.7000 0.2710 7.29
2500?) ee eae 003 0.2420 6.70
ILGEIE AWAY eet ey Beal een 1.8505 0.1085 5.34
PMT) Susie es ca ichc, te xe ge at 1.8385 0.9840 4.56
a (UC) Seen’ _ . S270 0.0820 4.49
3. Determination of lecithin by A. Schulze’s method.
Dry matter in grms. Lecithin in grms. Percentages.
TNOOE (Aue = oo atee 20 tee 3.6360 0.03635 0.999
=ssgpts( (23) RI ae ea i 3.8048 trace
1526 al (09 eae: set eee ee 3.7160 0.03998 1.076
MA (3) Red ee ee Gk eore thee 3.6088 trace

4. Determination of total carbohydrate.

Total carbohydrate

Dry matter in grms. in germs. Percentages,
LK (CCOEL(2'3 he Se oo ee ae 1.3635 0.2050 15.04
nce) A) oie A aa ar act a 1.9025 0.5048 26.50
LS BR 2((0') ae ee ae 1.8580 0.3678 19.80
BMPR Sho. on wlohe he 1.8044 0.3152 17.20
LA! (A iste a 1.3878 0.3212 Ta!
FP) ieee ole ave. as oe sO 0.6320 22.92
PC) tisdc swiss «/02 » sscete Dees 0.6424 23.44

5. Determination of sugars.

Dry matter in grms. Sugar in grms. Percentages.
LACM AUC heehee si aicies. oes 4.5450 O. 3004 6.60
“3 NYS) ae ae Os an ee 1.902 0.2010 10.50
1B E2(0o') Ee ae Pee a 1.8580 0.2490 3.30
noi) 2) at oe EE a 1.8045 0.1640 9.07

All sugar determinations were made by A//hn’s method.

498 MAENO.

6. Determination of total nitrogen.
Dry matter in grms. Baryta-water in cc, Percentages.

Root. (0)... ae ee 1.8180 12.8 Enz
7 GB) eae ee eee 0.9512 3:5 1.150
Bark (Qc. 25.2 See 0.9290 [a 1.650
S208) ie eee pie 019022 4.6 1.563
Ikeat(A))) ..s25.ne eee 0.9252 12.6 4.250
(B)) a;t its chee 0.9192 II.0 3.730

(Cy. sce. See eee eee 0.9135 10.9 3.720

7. Determination of proteid nitrogen by Stutzer’s method.
Dry matter in grms. Baryta water in cc. Percentages.

Root(@)) 2... 35 26 ee 0.90g0 3.8 0.898
55, SBA: Scene ee 0.9512 1.2 0.304
Barks(@). <. 22265: see 0.9290 5.6 1.294
(By. cxckeiee See 0.9022 3.0 1.040
eal (Athos ane 0.9252 18.2 3.470
(B)> etowen oe eee 0.9102 9-7 3.290

(CG) Sactete stare 0.9135 9.6 3.280

8. Determination of asparagine nitrogen by Sachsse’s me-
thod.

Dry matter Baryta water Percentages of

in grms. in cc. nitrogen x 2.
Root (@) ...°5.4065 <2 oc 4.5450 5.5 0.520
3) UP): oie 2.8536 3.0 0.656
Bark (@) 254 <2... eee 4.6450 3.8 0.351
(P) en oranee se oe 2.7066 oA 0.550

In the case of root (a) and bark (a) I cc. baryta water
=2.14806 mgm. nitrogen and in the case of root (f), bark (A),
leaf A, Leaf B, and leaf-C, Icc. baryta-water=3.12445 mgm.
nitrogen but only in the case of leaf A in proteid nitrogen deter-
mination I cc. of baryta-water=1.77115 mgm. nitrogen.

Notes on the Metabolism in the Cherry Tree.
BY

S. Aoyama, Nogakushr.

Numerous investigations of Sachs, Node and others have
shown that the bark and wood of trees store up during the autumn
a considerable amount of reserve-material which is transported
there from the leaves and partially prepared in the living bark
itself. This material not only serves for the nutrition of the cam-
bium but also is partially transported to the buds of the leaves and
flowers during their development in spring. Robert Hartig has
found that in the wood of the trunk of the red beech and oak,
starch is deposited toa considerable extent. With old trees of red
beech the fifty exterior annual rings contain reserve-starch which
is used up in those years in which seeds are produced. In other
years, however, only the last two rings show in summer a decrease
in starch, while in October starch is again deposited in them.

This author therefore infers that for the nutrition of the leaf-
buds in spring the reserve-material from the branches is mainly
used. Rudolph Weber has especially examined the rate of de-
crease in proteid and mineral matter in the old and young parts
of the trunk of the beech tree during a year of seed production,
and has found that a large amount of nitrogenous matter and
magnesium salts migrate from the wood to the growing seeds.“’

Russow and A. Fischer found that conifers store up the re-
serve-material free from nitrogen principally in the form of fat,
while many other trees, especially hard wood trees, store it up in
the form of starch. The amount of reserve-material, further, will
probably vary under different conditions, especially in different
climates.

It is an interesting fact that in the central and southern parts
of Japan the cherry trees generally bear no fruit” and in certain
cases in which the development of fruit takes place the fleshy
part remains imperfectly developed. On the other hand these
trees show in spring a most luxuriant development of flowers,

(1) Dr, v. Tubeufs Forstlich-naturwissenschaftliche Zeischrift, vol. I. (1892).
(2) Only one variety forms an exception.

500 AOYAMA ;

which form a dense cover of the branches before the leaves
appear.

It is evidently the peculiar climatic condition of central and
southern Japan which prevents the production of normal cherries
and causes the fruit to fall off in an unripe condition. This cir-
cumstance must naturally lead to the accumulation of a great
amount of reserve-material in the bark and wood, that would
otherwise have been consumed by the ripening fruit, and this is
clearly also the cause that leads to the development of such an
extraordinary and astonishing abundance of blossoms in the fol-
lowing spring. I therefore believed it of some interest to deter-
mine the amount of reserve-material in winter and to compare it
with the extent of the consumption of this reserve-material in
spring when flowers and leaves have been formed.

Branches I—1.5 cm. in thickness were collected on the 20th
January (A) and, again of the same tree, branches of the same
size on the 13th April (B) on which day also I collected from the
same tree young leaves and flowers. .

Only the leaving part of the bark, containing more or less
cambium, served for analysis.” The results I obtained are the
following :—

Bark (A) Bark (B) Leaf. Flower.

Total water... 4.2.) eee; 52.64 53.41
In 100 parts of dry matter.

Crude "protein 4.0... 152 eee - 9.50 6.69 34.94 ZO
Crudé fat) 2) 2 ice eee - 6.84 5.34 8.99 10.04
Crude fibre... 52... aa. ae. 34.79 38.98 14.49 20.06
Carbohivdtatem eee me. 27.13 18.06 11.82 12.50
Crudesash:. <2204¢. 0 eee - 776 7.93 6.51 72s
Non-nitrogenous extract ..... 12.46 21.93 17.66 26.80
Total nitrogen*..7- eee Eas 1.07 5.59 3.22
Albuminoid nitrogen 2) ee. 1.16 0.84 4.55 2.40

If we compare the precentage composition of bark (A) with
that of bark (B), we can obtain no clear distinction between re-
lative and absolute numbers, zc. between the apparent and real
decrease or increase of the different constituents; evidently a

(1) The wood was not examined. Some trees contain more starch in March and
April than in January (Rosenberg, Bot. Centrbl. 66, 337). Cf. also the above examina-
tion of the root of the mulberry tree, by Maeno. It may he that in such cases fat and
proteins contributed to the formation of starch.

NOTES ON THE METABODISM IN VME CHERRY TREE. 501

more correct estimation as to the changes can be obtained if we
take the fibre as a constant quantity, which may here safely be
done, and recalculate the numbers for proteids, fat, and carbo-
hydrate. We obtain now the following results :-—

(A) (B)

FE iDrei eee cle. soe =. 100.00 100.00
Grude, Proteid:. 7s accom ce 2730 17.16

FOC a eee: .. an 19.67 13.70
Carbohydrate. 2... ..seee: -« - 77.98 46.33
lencerthe:protetds decreased ........ seamen ee. B7 slog,
atidecreased. 2 tenn: -.... «4.5 aera 5 205355,
Carbohydrate decreasediiem.... --.: .. merase en 40559),5

This shows that the bark of the twigs plays a very important
part as a reservoir for the development of the buds in spring.
Analytical Data.

Determination of total nitrogen.“’
Percentage of

Dry matter, g. Baryta-water, cc. nitrogen.
Lares) (VAN) aes ee 0.940 6.70 1.52
cgi (18) lee, Saas ie 0.8¢0 5.30 ACY
Westerner nda tet sauce. oe 0.88 3 27.90 5.59
PPV Cli. Vacve er shane oss es eos 0.879 16.00 Bi22

Determination of albumoid nitrogen by S7utzer’s method.
Percentage of

Dry matter, g. Baryta-water, cc. aihyeereay
| ysl) (2a) see ONCE, SPER Bice eee 0.946 5.40 1.18
no 18) ahs eee eee . 0.8g0 4.20 0.84
11 (SRBF sete oe oS ERR en ae 0.883 22.70 4.55
PONE Te eesti a sia) a te ee nes 0.879 16 Golo) 2.40
Determination of crude fat.
Dry matter, g. Crude fat. Percentage.
SAGAN ena sts chek tee sual ee 4.728 0.3235 6.84
nj OA LO he Ne ee EP 4.449 0.2377 5.34
ILENE ct eR to ae ae ere 4.415 0.3970 8.99
1 SUE Ge ec ok, CERT Rec 4.390 0.4415

10 04

(1) All the nitrogen determinations were made by A7e/dah/’s method.

10 cc, of sulphuric acid were put in the receiver.

In the case of bark (A), 1 cc. sulphuric acid =3.16 baryta-water solution, and 1 cc.
baryta-water = 2.14806 mgr, nitrogen.

In the case of bark (B), 1 cc. sulphuric acid=2.16 baryta-water solution, and 1 cc.

baryta-water =1.77115 mer. nitrogen.

502 AOYAMA.

Determination of crude fibre.

Dry matter, g. Crude fibre. Percentage.
Bark (A). vac. shinee 4.728 1.644 34.79
wy. CB) era a 2.070 1.041 38.98
Leaf ..shslccs nee 2.649 0.381 14.49
PIOWER. 5.4.5 i244 on eee 2.637 0.529 20.06

Determination of carbohydrate...
Total

Dry matter, g. carbohydrate, g. Percentage.
Bark (A). .... 1255 se 4.728 1.283 27.13
CB) 2s..3* Lo eee 2.670 0.482 18.06
eat onc. Pt.. Se 2.649 0.313 11.82
FlOWeéf.. 3.238. 2 eee 2.637 0.330 12.50

(1) This determination was made by the acid-alkali method.
(2) This carbobydrate consists of starch and sugar; the determination was made

by 4d/hn’s method.

Physiological Observations on Lecithin.
BY

T. Hanai, Vogakushi.

It is a well known fact that lecithin occurs widely distribut-
ed in the vegetable and animal kingdoms, forming in various pro-
portions an admixture with fatty matters. Only a limited num-
ber of observations have however been made in regard to the
physiological relations. It was found by Maxwell,” that the
.amount of lecithin increases during the germination process of
plants, and later decreases again. S. Frankfurt observed that
during the germination of Helianthus seeds the amount of leci-
thin increased from 0.44 to 0.85 % while the amount of fat de-
creased from 55.32 to 24.54 %. It seems very probable that in
reality much more lecithin had been formed during the germina-
tion process than was actually found, and that a part of it again
was consumed.

O. Loew made some experiments with a diluted solution of
lecithin in regard to the capability of nourishing lower fungi, and
observed that Pentctllium could not, in the absence of other or-
ganic material, develop inao.1 % solution of lecithin containing
the necessary mineral nutrients, but only bacteria to a moderate
extent; this vegetation made the impression of a pure culture,
although the infection was made from putrid meat containing
various kinds of microbes.“

Seeds rich in starch generally contain much less lecithin
than such as are rich in proteid, thus barley grains contain less
than half the amount of lecithin that soja-beans do.

Probably there is also a larger proportion of lecith-albumin®
in the seed of soja and lupin than in those of squash and barley.

(1) Chem. Central-Blatt., 91, I. 365.

Hefter olyserved a decrease of the amount of lecithin in the liver during starvation.

(2) Landw. Vers.-Stat., Vol. XLII, 143.

(3) Recently Stok/asa (Wien. Akad. Ber., 1895) has found that lecithin forms a suit-
able source of phosphoric acid when offered to the roots ; he also observed its formation
in green leaves as well as its consumption in darkness.

(4) Bull. Coll. of Agr. of the Imp. Univ. of Japan ; Vol. II. No. 2.

(5) Schulze and Steiger, Zeitschr. physiol, Chem., 13, 386.

(6) Cf. Leo. Liebermann, Pflitg. Arch., 1893.

.

504 HANAT;

E. Schulze found further that during the germination of seeds
the quantity of choliz increases, and that in wheat cholin and
betain which are closely related to each other, are localised in
the germ of the grain but not in the endosperm, This is certain-
ly of physiological interest because the young developing germ
must Carry on an energetie respiration and therefore be capable
of easily forming lecithin, in which process the presence of cholin
is required. It may further be mentioned in this connection that,
according. to Mintz, the amount of free fatty acids increases
during germination.

For my investigation I selected the leaves of Thea chinensis,
an evergreen plant, and the bark of Prunus: Cerasus, which ob-
jects contain during winter time much reserve material ; especial-
ly the tea leaf is rich in fatty matter. Kelluer, Makino, and
Ogasawara” observed that while in May and June young tea
leaves contain 6.32-6.82 % of fat, in November they contain as
much as 22 % inthe dry matter. I determined the lecithin after
the method of &. Schulze by which after the extraction with ether,
a second extraction is made with absolute alcohol.” In these
united liquids the phosphoric acid is determined in the usual way,
after heating the evaporation-residue with a mixture of sodium
carbonate and some potassium nitrate.

I made. four determinations, one with the o/d leaves in
November (1895), the second with the o/d leaves in May 1896,
the third with the young leaves at the beginning of April 1896
and the fourth with the young leaves at the end of May (1896),
with the following result :

(1) Landw. Vers.-Stat.. Vol. 46, 23.

(2) Landw. Vers.-Stat. 1886. p, 370.

(3) The mixture of these two extracts contains, of course, besides fat and lecithin
a certain amount of other compounds.

PHYSIOLOGICAL OBSERVATIONS ON LECITHIN. 505

Watesaathe In 100 parts of the dry matter.
No. Dates of collection. fresh leaves,
per cent. Ethereal and eae
alcoholic extract. Lecithin.
Old leaf
T- | 20 November (1895) 67.57 26.18 2.54
Old leaf
2 26 May (1896) 61.88 18.19 oO
Young leaf
3: 1 April (1896) 77.06 9.44 0.21
Y leaf
4 26 ar 1896) 1252 18.67 1.11

This result shows that the old tea leaves lose the reserve-

lecithin in spring, while the amount of it increases gradually in
the young leaves. The decrease and the increase of lecithin here
goes parallel with that of fat although not in a fixed proportion. -

The bark of Prunus Cerasus was collected on the 23rd
October 1895, when the leaves had mostly fallen from the tree,
while the second collection was made on the 5th April 1896 when
numerous flower-buds were formed, and the third time on the oth
April 1896, when the flower-buds had opened.

In the three cases the bark was taken from the same tree
whereby special attention was paid, that branches of equal thick-
ness were Selected ; the results were as follows:

| In 100 parts of the dry matter.

No. Dates of collection, |
Ethereal and alcoho- Lecithi
lic extract. =
ema le2 SO ctober (LS95))) +..,maseeeee | 10.53 1.88
Zz BEADLE (TSQG)) sn. saateen et eer nee meee: | 10.97 0.96
3: QRAPTUNCLSOGOS |. dacnsss acest eee, | 9.52 0.71

nnn een FR

It is also here quite evident that lecithin is a reserve material
which is consumed in spring.

506 HANAI.

Analytical Data.

For the determination of the lecithin always 10 grm. of dry
matter were taken; the amount of magnesium pyrophosphate
found was as follows : =

Ethereal and} Magnesium

alcoholic pyrophos- Lecithin,

extract. phate.
Old leaf of Thea chinensis. 20. Noy. (1895). 2.618 0.035 0.254
OlditeaileahS (26) Mayi(1896) eaneennee teeter - 1.819 == --
Young tea leaf. 1 April (1896) ............... 0.944 0.003 0.021
Young tea leaf. 26:May (1896) ............... 1.867 0.015 O.1II
ee AR
Bark of Prunus Cerasus. 5 Apr. (1896) ... 1.097 0.013 0.096
Bark of Prunus Cerasus. 9 Apr. (1896) ... 0 952 0.009 0.071

On a Compound of Albumin
with Phenol.

BY
M. Shimada, Nogakushi.

Finely powdered dry egg-albumin dissolves gradually when
heated with 10 times of its weight of phenol for several hours on
the water bath. From this solution alcohol precipitates a floccu-
lent mass, which, after washing with alcohol and water, repre-
sents a compound of albumin and phenol.”

This compound is without taste or smell, insoluble in boiling
alcohol and water, and in solution of potassium carbonate, easily
soluble in hot phenol. In concentrated acetic acid it swells up
gradually, while potassium hydroxide, even in dilution of 0.5 per
cent, dissolves it; from this solution acetic acid precipitates it
again.

It is not attacked in the cold by hydrochloric acid of 10 per-
cent, but gradually dissolved by one of 35 per cent. Nitric acid
of 5 per cent has no effect on it at the ordinary temperature, but
on boiling with concentrated nitric acid, a yellow colouration is
obtained. It gives the biuret and JZ7//on’s reaction like common
albumin.

I germ. of this product was digested with 20 cc. of concentrated
hydrochloric acid at 100° C. for several hours, and then the liquid
was subjected to distillation in order to see, whether phenol was
hereby liberated, but the distillate obtained did not give a trace of
turbidity with bromine water. The same negative result in regard
phenol was observed in the following experiment, which was made
to see whether that new compound would also yield leucine and
tyrosine, like the common albumin.

I heated 3 grm. with 15 cc. of sulphuric acid of 30 per cent
for seven hours on the water bath and then for a short time on the
sand bath.

(1) If the amount of alcohol is too small, then instead of a flocculent mass, a tenace-
ous mass is separated, from which the adhering phenol can be removed only by prolong-
ed washing. (2) Peptone behaves similarly to albumin, O. L.

(2) This did not show the biuret reaction but yielded a very copious precipitate
with phospho-tungstic acid, pointing to a considerable amount of basic compounds.

508 SHIMADA ;

Upon dilution with water, boiling with barium carbonate,
evaporation of the filtrate and extraction with alcohol containing
some ammonia, a characteristic crystallisation of leucine and
tyrosine was obtained. The leucine was also recognised by the
odour of amylamine on heating, while it showed tyrosine by
Millon’s reaction.

Whether also arginine, lysine, aspartic acid, glutamic acid
and phenyl-amido-propionic acid was formed, I hope to decide
later with larger quantities.

For analysis the product was dried at 100° C.

I. 0,230 grm. gave 0.4863 grm. CO, and 0.149 grm. H,O.

=5 705 % C and 7:2 % H.
II. 0.1636 grm. gave 0.3500 grm. CO, and 0.103 grm. H,O.
=—meee % C and.7.0 9% H:
I. 0.5000 grm. gave after A7e/dahl’s method
68.03 mg. N=13.60 % N.

II. 0.5000 grm. gave 67.22 mg. N=13.45 9% N.@

I. 1.0000 grm. substance gave after heating with a mixture
of sodium carbonate and some potassium chlorate 0.117 grm.
BasO,— 1.60% s:

II. 1.0000 grm. yielded by the same method 0.090 grm.
BaSO,== 1.30.9) 9;

These results would correspond approximately to an albumin
in which three hydrogen atoms have been replaced by three pheny!]
eroups, When Lieberhiihn’s formula is taken as a foundation.

The product must then have been formed according to the
following equation :—

Cp Ay2NisSOx + 3(C35H30 A) = C72 Hy9(C5H5)3N1sSO.2 + 3 HO.

Experiment.
Theory.
if Il.
Cc 58.69 57.65 58.34
H 6.74 7.20 7.00
Nie 13.70 13.60 13.45
SS) 1.74 1.60 1.30
O | 19.13
100.00 |

(1) Another sample yielded Swzk/, of this College, 14.44 9 nitrogen.

ON A COMPOUND OF ALBUMIN WITH PHENOL. 509

I prepared the compound several times and always observed
essentially the same properties.

As it appeared to me of some interest to test whether this
product would show, in the absence of air, antiseptic properties on
account of the introduction of phenyl groups, I dissolved 1 grm.
jn potassium hydroxide solution of 1 per cent and added dilute
acetic acid until a precipitate commenced to be formed, diluted
to 100 cc., and infected the solution from putrified meat. The
filled flask was provided with a stopper, carrying a U-tube con-
taining some mercury to exclude the air.

After seven weeks standing at the ordinary temperature,
the liquid appeared turbid and contained a flocculent sediment.

Upon opening a putrid smell was noticed and the microscope
revealed a rich bacterial vegetation.

Triphenylalbumin is therefore a good nutrient for microbes
and is subject to fermentation like the ordinary albumin.

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