[Pe fe saad WIE J 10} — .25
MEPs fo be ne Le teadaltestoale clactocloale ale clooles 714s (ea BA ARV | SS 6 ER 6 0
Gee 8 1M As PRE ae) Ea Es ye | 8} + .25
(ae Lt Al Sto The ee eR ie EE ee oe
WD ac. Me). ae Boh ie 6} + .75
oo pape) se | 2} +1.00
Bees: . a fs | ap.) ee 44 +1.25
8.502. weeeee eee aH eee eee Ee 1} +1.50
Dregqueney.ci. 52.00.22. 06 02: 1} 0} O} 1} Of OF OF 1) 1} 1} 3} 4) 8} 8 5) 7} 9} 6 4] 5) O} 1] 1) 66
SIASIRRIRSIES[alsleia/a| [Risicisiaisiis|
from Mean ............. OD JOD LOD ONION ICR i |i Ti ; IN
ees” Fost ean PEPEPOPTPAPTPTPT PUTT] a] 1 UP | eee
USE OF CORRELATION STUDIES IN BREEDING WORK.
- The value of studies in correlation to the practical work of plant
improvement, while not questioned by those familiar with statistical
methods, has been doubted by those not in the habit of doing accurate
263
36 BREEDING ASPARAGUS FOR RUST RESISTANCE.
work. One or two examples will show the possibilities of the use of
correlation. |
The relation of the size of the asparagus stalk in the fall to the
next year’s cut is interesting, as’ it is necessary in selecting rust-
resistant plants in the fall to pick those that will give large-sized
shoots. Studies of several plants in row Al presented in Table X
give a fair idea of the value of large-sized fall growth in determining
large-sized spring growth. In the same way the total yield should be
taken into consideration. This quality is hidden at the time of
selection and must be correlated with total production of stalks in
the field in the fall. This relationship was worked out with the
plants in row Al and the result.is shown in Table XI. Certainly when
the correlation is as high as 0.8 the observer should make an almost
perfect selection by using the correlated character to pick the very
best plants.
TABLE X.—Correlation between the diameters of the largest stalks in 86 hills of row Al in
the fall of 1909 and in the largest stalks cut in the spring of 1910.
[Diameters of 1909, subject; diameters of 1910, relative. Coefficient of correlation 0.575+0.050.]
Spring of 1910 (}-inch units). De-
Fre- parture
Fall of 1909. a i
. 2/314|51]61{7{81] 9] 10] 11] 12] © | mean.
pag A apapentcriee! 2) LS TR ed (oe ha pee He Ae ON tees fe Ye —2
BRD ae ge OF ee ne ee Fy eames es ES FE er 20 ae
tei ty Seay ee ae 1} 4] 3] 6] 42) 6] 11... eee 32 0
hy SUR Ge ees VER Pane 1] 2403) 5457) 4) 1 ae 23 +1
“PRES Pekin SE ae RS A) FG ee EP. 91”, Ba od 1.02 te 7 +2
i a OS Sa gt SEE -¢ anion i VR Gen as 2 9 Sich! Me +h. Biron |. a 3 +3
Braiaomne.. . J--4.-0-t6£ 2)... --- 2] 6|14]16| 22/16] 8] 1| 0| Oj 1] 86
Departure from mean... -. 5.2. --...-- —4 |-3 —2|-1j; 0Oj+1 [+2 |+3 |+4 \+5 eee
TaBLeE XI.—Correlation between cross section of stems in fall of 1910 and cut in spring
of 1911 on 82 producing plants of row Al.
[Fall stem area, subject; spring cut area, relative. Coefficient of correlation 0.8114 0.025.]
. i-?) .
Spring of 1911 (square inches). e 3 a
_—
Fall of 1910. 5 | 48
1/3/5117 /9]{11/13 2g Ba
B&B |/A&
j-inch square units:
pis SERN Se a ee alk: nh =
NR Mi Set Oe RE 23 6 6 7 os |
Dib sok nalt ad kas ae ge es 16 0
WL sch winic ¥en, slot iret ole abs - - i 4] +1
O0a 0 Leek ee ET..|...- 4} +2
A ee Fit ores es) Sy hae ore 4) +3
190;.0.5.48ct dames ne) od $(<3-8
Mts cad as venders. - . 2| +5
ER Ra Fes aa . 1] +6
REA SH \. 0; +7
TAG cos orcs kn cde SRE MMLC tx «| =< [a 0's chy vals pel aaicwelec ale dl: 2 alae Opbimaia coe 1k aan o| +8
BOOK strc wish cog Chee a | op a[ensafeucfesel Bale. wallccclaw alivaehewelli ahaa ca enna 0}; +9
9 aS Se RTT 10 Liat TI | OF Fee Fs les Be» He 1} +10
FAR Mis uy esd enet ekut ade RT ee fered Sap" fata Sis ts Re ede 1] +11
Frequency........-. 16} 14) 13
Departure from mean..... .
ee
BREEDING. | 37
A most mteresting interrelation comes in the size of the seedling
_ to the weight of the seed. It was early noticed that the greenhouse
lots of seedlings showed striking differences between progeny lots
from different female plants. Wherever enough seed remained
unplanted to give a fair average, the seed weight was determined and
a comparison made with the average height of the seedlings in the
progeny rows. Table XII was obtained with a coefficient of 0.780.
Where future size depends on the start the young seedling gets in
the bed, the tremendous importance of the use of large seed is at
once apparent. To further test this correlation, 100 seeds from plant
C13—5-1, open fertilized, were sown in 1910 under uniform conditions
of moisture, heat, and light in a soil of uniform texture. Each seed
was weighed to 0.001 gram, the germination record was made, and
the height of each seedling was measured daily. No effect of size of
seed on germination could be determined, but the size of the indi-
vidual seed showed a very strong influence on the height and rate of
growth. (Table XIII.) Where the individual seed is taken into
account the correlation is lower than where the average of several
is taken, on account of the varying hereditary tendencies in different
seed of the same weight.
TABLE XII.—Correlation between average weight of seed and average height of green-
house seedlings of 1910 for 42 progeny lots on February 11, 1910.
[Height, subject; weight, relative. Coefficient of correlation 0.780+0.042.]
Aver ight of s milligrams). De-
erage weight of seed (milligrams) ie: pare:
Height of seedlings. a a a ih tel CPOE:
ey. | from
19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | 29 cease
Inches:
POM etcetera ao MSM NS ih gli cmPardc DSF Webel 2 ers level cose dtc Oe |e eget eter 1| —1.00
Meera oc nice Few eee okt 1 Le a a 2 Le MA a 5| — .7
NS SS ES SI ee a age ee Pee) | ea TS] Sea PSE SRE Smee ey, OP | pe We mae 2| — .50
ICE eh oes as SAS Sh tues ri) a ee eo a) Seas ee #y 95
he eh Se eS Sate ARR An ee ee rs (am ge Ra LF 2) Pe Wl | ee 9 0
EM ee ae OY Ne eo e GAEL No safe ae | eal: <2)? lS i a cae 10| + .25
Oo ea eee ae eee 1 (Re Pete gO Ne ee i 2 + .50
EMCEE Na Sc fe SU el ECE A aS Pele te ye Aa Ree Eleeiee P24 ee ee 5 + .75
OO cn TS Sp SADT aa ie | | ope get [Ona ele EO Pe bh aes 1 3} +1.00
LD case a i kes ee as SAT BE Teg ET (2) Uhh ho a 1 es ee ee 1 +1.25
121 20 2 1 (Re ee 2 6 9 | 4 | 5 Os | 42
Departure from mean................... Ea eG ORR eer Ofer +e Fe aa +5 +5 |+6 = Bathe.
263
ir? a
- ie
a
j a
4
3
38 BREEDING ASPARAGUS FOR RUST RESISTANCE. |
Taste XIII.—Correlation between height of shoot and weight of seed in 100 seedlings
from weighed seed of C13-5-1, open fertilized, grown under control conditions in green-
house at Washington in 1910.
[Height of first shoot April 9, 1910, subject; weight of seed in milligrams, relative. Coefficient of correla-
a eer
tion 0.41+0.056.]
ie! :
Weight of seed in milligrams.
-
Height of shoot. :
12/13] 14]15} 16 | 17 | 18 | 19 | 20 E 22 | 23 | 24| 25 | 26 ,
Inches
he a ain cha crete pal cea 1 oor caer me eae eee
qb okies Saabs 1).
Boe bar dn eg cnesnoc peNCoiie Gs bie yaa pees ERR ET | RS A
Spy.) NS fare SB Be eS eS 5 Se 23 2 1 He 3} 1).
Se aC ae BER pe eet aes 2} 1 hnu--1 oak: SL ee ee
WAGs St iw n onan SON Moen ebse [telat (ie ee ee oe Ge ee et Re
| Re ee ene Sa See ee” Pe ee eee re | me Nae: Wt eat | Paes fate
“hE ABE eS Pee athe Soe oe RD Oe Be iE. ARSE iL ie | a oe |
ES TE SE ere ee See ae ae ee re at | eee ee) Pa |
G16. [st AE eR : | eee 2 | 3
ERE Fe ee Se ae oa, CF
pf oe eee BS ae Ske d ee eee g :
ADO nao ats Side Su een ge ee et 1 ,
iy: Se SRS 8 SOF poet ee Phe 051 7 hoe 1 1
Mredueney = o25.5 seen cee 2} 3] 2) 7} 13) 12) 12) 10) 12) 9} 8 5| I} 2 2
Departure from mean............ 4-8 —5|—4) —3) —2} —1) 0} +1/+2/+3/+4/+5/+6/+7
Table XIV is introduced in order to show that the size of the young
asparagus seedling is important in determining the future size of
the plant. This table shows the correlation existing between the
average heights of 85 progeny lots while in the seed flats in the green-
house at an age of three weeks and the average of the same progeny
lots in their permanent place in the field. The factor of crowding,
which aids the stronger plants and retards the weaker ones, was
eliminated by placing the plants in individual pots February 13
and so maintaining them until they were transplanted to the field
where they are just beginning to show the effects of crowding in the
fall of 1911.
263
BREEDING. 39
Taste XIV.—Correlation between average height of greenhouse seedlings in progeny lots
February 11, 1910 (in 4-inch units), and their average height September, 1910 (1m inches).
[Height in September, subject; height in February, relative. Coefficient of correlation 0.552+0.048.]
Average height of seedlings February 11, 1910 (}-inch | ¢, = =
units). S eo
Height in September. 3 5, - ;
20 | 21 | 22 | 23 | 24| 25| 26 | 27 | 28 291301 31 32 | 33 | 34 & A&
Inches:
8.2 bine atte o gla t tunics aol Pae Bee Beech: Fete et ed DES ME 9 2) —6
od Zee ee ee ee cee See 1 aifaree arate bo PE Re Se ee ey eee, Beye 3} —5
Ln Pa a EASE ah: Oe Sere nee aera) Urges iN aes | Ba Dita spacchoha® cbs 6 —4
n+ cee CES SAR GER Sei Some ay Seca |e | Yea (ie | fue Zio las Supases £15 Se RES eS 44 —3
Co 2 ES RE a ees Seem ae eee | ee cl 3} «3 Abe Becks 14 —2
te SES SRE RES Se es Soper | aes | ten a OR ee | Be eee 12 abr | |S Paes MaRS vis 8} —1
Oo ge SHAG EES: SEAS Se ie ener | cred ed ae | er b | ees 2 Le? Shy. dhs Bho ees I: -12 0
LA Se RS a ae ey ee eee: a A pope ee Se 1 eS 2 i Soke Ape Bb Peo Eh 13 +1
SE tote goatee ao Smale ible o aps oe le eebeee Desi. sls. yes | ie ge) oe 7) +2
Mi rnd sek toe ih a io fm oft aaloe apa 6 Ueelt- wal dea 1) a a eee Pe Us soe 3} +3
TS I ot Bek ol. Loc as Seis A St <[b dole s dlemelOantews tI oe se ae a Ue | | ae | ee 6} +4
1 he GEC SIE BRR Se as SE Se epee Dea (ne Me | em ed lo el Oe ee a Tio dies seeetaaeee 2) +5
oc cyl RRR FR Rr ane Seas Sgr Se ee | ee Ee ree) 1 Soa} pA RRS ew 3} +6
Ss LES EED Sieh GRRE RES SEE NM GT AE oy ee Gl ee Bee 1}...]--.|---[---| U---] 2) 47
1 a a ee a 1; Oo} 1) 2) 8 2 10) 15) 17) 6 8 6 5 3) Ye 8
Departure from Mea. eee eeeeeese-| “876-5 —4—3 ae —1 O41 +2+3)44)45)+9)....
In continuing the work with the greenhouse seedlings after they _
were removed to the field in May, 1910, further correlations have been
studied. While these progeny lots are not exactly a random sample,
in many respects they answer the purpose of one, the population being
distributed in monomodal and often nearly normal variation curves.
In making selections among the young seedlings a general tendency
among growers is to assume that height is a good index of large-sized’
shoots in future years in the cutting bed. According to the tables
constructed on the data of height in 1910 and stalk diameter of the
greenhouse seedlings in 1911 (Table XV), there is a correlation of
0.634 +0.013. This result is supported by notes from row Al, where
the total area of the cross section of the fall stems in 1910 was com-
pared with the total area of the cross section of the shoots.cut in the
spring of 1911, as shown in Table XI (p. 36).
263
40 BREEDING ASPARAGUS FOR RUST RESISTANCE.
TABLE XV.—Correlation between height in 1910 (inches) and diameter of largest stalk
in 1911 (25-inch units) of the greenhouse seedlings of 1910. (
[Height in 1910, subject; diameter in 1911, relative. Coefficient of correlation 0.6344 0.013.] 5
Diameter of stalks in 1911 (#g-inch units).
Height in 1910.
1)/2131/4/)6 164 74879 11011] 18) Bie
Frequency
parture
om mean.
-
_
o
:
‘
‘
‘
‘
'
'
.
.
eS Va = See
Peequeney 22: fo... Fe:
Departure from mean
Table XVI shows a comparison of height of the greenhouse seed-
lings in September, 1910, and in 1911. This table serves as another
illustration of the opportunity of making selections on seedlings.
Seventy-five per cent of the 1910 lot could have been discarded with-
out losing any of the 10 best plants of 1911. A comparison of the
correlation here shown, with the table presented by Clark?’ in his
studies of timothy in 1910, shows that the conditions at Concord
were more uniform than those at Ithaca.
One striking feature in connection with these studies is that the
reliability of the height correlation is almost equaled by the relia-
bility of height with next year’s size (Table XV).
Data from mature plants in row Al show a high correlation
(Table XVII) between the amount of tops on the plants in the fall
year by year and also a high correlation (Table XVIII) between the
cut of individual plants compared in successive years.
1 Clark, C. F, Bulletin 279, Cornell Agricultural Experiment Station. 1910.
263 i} :
—
> ae
BREEDING. 4]
TaBLE X VI.—Correlation between heights of the greenhouse seedlings of 1910 in 1910 and
in 1911 (inches).
[Height in 1910, subject; height in 1911, relative. Coefficient of correlation 0.642+0.0126.]
. om .
Height of plants in 1911 (inches). | 5
a =
Height in 1910. | [| | 5| %&
3 | 6 | 9 /12,15)18/21/24/27|30|33|36 39] 42 |45 48 51/54|57/60/63/66|69\72|75|78|81184| © 5s
a &
nches
‘ eee ees al fa Peo ON LP 7 ee a) |G al a ha) al ae Pa ey 2) —ll1
SE See 1 + AN (Oe i 8 eae FR ss PE a ee pe _.|..|.-| .5}. —10
I Se ae id a Sich Sra. to cP Yt ot Bl ste hs oho tle alas: bee teak ..|..|--| 12} —9
SS Sa ae Oe ee fe SA) Ho-b 2n-t Gr Gl 4ieart A et a. he bk _.|.-|--| 29} —8
ee te the, 1} 2} 1} 3} 6] 2} 6 1 6) 1) 2)..)..)..)..|-.]--J--. _.|..|..| 31] — 7
peer ate eel Tented be 1| 5} 8} 6] 811) 4) 7) 5) 1) 1) Qt. ef. ef fee] ..|..|--| 59) — 6
et eae Biel id ae) Re 2) 3] 1] 4] 6] 7] 3i1t} 5) 5] 1) 2)--}.-|..}..[. fe. .Jasf--f SOpu— 6
ERT aS ee ait Sie Oe 1} 2} 2} 6/10} 5/12) 9] 17] 3) 3! 3] 3] 2] 1)..]..]..]- ..|..]..| 79} — 4
ee OO Rea PRS id ee a Se ae 1}..} 4110} 817/11) 9} 5! 6| 2) 4i..J. 1. pooped. - [onto te GR 2
RNG Git rae ae Fe ae ee HG DP 2) 2|..) 7/14] 5] 17/10] 8] 3] 7}..|....]..]..]..].-].-|--|..| 75] — 2
eS OCEAGIS SRSAER I AS ES RE CS ee a 1} 3} 4} 6] 5)12) 15] 9] 6) 9.11) 2] 2/._|_.]..|. ..|.-|--| 85) — 1
ESR eee Pi a 1] 1| 2} 7[14) gfi4iz4a: 4} 8] 5]._f. 2]. .|eefe.].ff.| 79 0
ee ee ee ee a hee laches ste 1} 1} 2} 3] 3) 8} 9/10) 81310} 2} 1] 2/.-|..). sch ee 2
ee te look oleate toot dt Sh tobe OF 70! 8) OT St. 2t..t tT _.|.-|..| 56) + 2
eee oe SSF ah ete tele Ie of AF Sb 8} 3) FOP 9112) S$} 6) 3) Biot. .f. I. ..| 60} +3
Sse hoe, oe tnd a ail blde ty 1) Sito! Sf Git Et. 1. -|. ..| 46, +4
me Le sal ci ‘ 1} 2}..} 3] 2) 4) 7| 5) 4) 3] 3)..] 1e.}..}.c}.-1..] 34) 45
y a he ell ek a GA 8 .|.-|--| 1] 2}..] 3] 5} 3] 5) 5} 5] 6}.-] 1]... i ie OF ey ae Sy
: : a - wel. tae St. ot 6) 4 Aa Ot Bie a ee a ee
cn hh xe ap) 2 2| 1] 2] 4) 4/ 2] 2! 3] 1] 2}. ..|.-]..| 23} + 8
i ies 4 F Rie wa) AI SL. SP at Hic: alt ll} +9
oo ae a a : SP OCS Ee) ee a ie i Lal 7; +10
5 V3 let PSPS eR sl OP RP Let [oe te ban ere
eh TS Sore le oa A Pe OR Sk a a tea aad Nel RO li Wt ie etl A sg) gba
ee Pee de Pea ahd te loakeefwole tected hecbtes : Bly ie ..|..[..} 2] +14
eree... e . Z ” 3 et ah 0} +15
ae sis : : 1 ea 1} +16
a Reece We ee OR oe eee SY
Frequency ....... 1} 0} 0} 5} 1] 7} 9/15!35/55169/84/91126/92/93/92|/83/42/28/12 6| 1} O} O} 1) O} 1} 949
Taste XVII.—Correlation between area of cross section of fall stems of row Al in 1910
and in 1911.
[Stems in 1910, subject; stems in 1911, relative. Coefficient of correlation, 0.859+0.018.]
Area oftross section of stems in fall of 1911 (3-inch square units). =
&
: { PB od
Stems in fall of 1910. aq -j/3s
a
cololololel[olo Sy) =e a= a re = me
SAS pete Se Stare se te tS tm te
4-inch square units:
_. 4. Sis Ole UES Sates See Meh bea Woe cl Ah Poe cle 5 18) >= 2
RErEners eer A ae Sha x Pe 5 Gi Anas abe Lee ails esa fata lterete pact urete : 18) —1
i 2 sie SERS SOG Sa a 2 5 se Aloe i Die Disa tata he o 16 0
FUL Ze SER TREE ae ha Oe rey hee a gs RE ie: | Ck Sa et el Me es Fa F 144 +1
ec ac keclels welaiah a [bdtole atalesatote Aa ed | a Dict ial eooh ss Sapecer Leese 44 +2
UL a) Se Se ree ne | Ye Oo Hye.2 1 pa | ae Py SO ate a 44. +3
LeU), - DS) a a. SS a A ae eee ee a US Pe 1 Brad voralotate - 3} + 4
ea re Lt eos Gs oft ok Wide fe. de] cee le cutie abeaatecs Asean obes coke aee 2} +5
eee LEE sD lca af adn ay ofa addadule aatvatheg sferdahatabc@les cit lela cocheaw. 1 1} +6
eee a Sate. Ms aloe Stee ciel tue ebs dab supdbulc alvictan ftecds conte cculaccd 0} +7
ee aa Racial ia «ufos cftne ofeeelesuf sbelaaabarshetatcd le Jatwuc antec ct wattodes 0} +8
ermeneeries ie eae Sa 2 es ooh anti dal eel awtl au akseafseultef dutecs|ecleseclessclecse 0} +9
ree ret). Pe ald, calc nae Mccabe swalu cafesctbeab ch dlc as be wel ance faut 1 1} +10
EE SS UE IRE) aoa | GORA RE | rsa ee | Sel ae” se Bec Wee |S a A Oe Tees 1} +11
Frequency......... LOPES LOA LOS 7 SO pea Dieser tl Ole Or - OL “Ae 82
Departure from mean..... —4| —3) —2) —1) 0)+1/+2/+3/+4|-+-5|/+6/+7/+8)]+9]+10)-+11/4+12).._._.
263
42 BREEDING ASPARAGUS FOR RUST RESISTANCE.
Taste XVIII.—Correlation between the yield in 1910 and in 1911 from the plants in row
Al, given in the sum of the squared diameters of the shoots from each hill.
[Cut of 1911, subject; cut of 1910, relative. Coefficient of correlation, 0.797+0.027.]
Cut of 1910 (square inches). 2
&
e |ed
Cut of 1911. = g s
- al+t}|olol|mloj|lea s aA
Z/SI/Z/B2j/8)/8 (81,8 {8 2 |e
olanlalalwi{o|]o]r |o & 1A
Square inches:
O40: 2s, Abit ep wactetiich as ese 16 |e cce sof ssl ae ee a oe ee — 6
TNA oe ele waren a betas kmiie bis lela alps T | Ob Bese Soles. ee ee eee — 4
PT is edie ae SES RIE RR are Diy ER SO Ee 2) GP B17. Ea Bs ee —2
oD Pe eee eee eee, pee ter Oo) BP Bete Bie ae Ce Rees ees ee 5)
STO 10... os Sento coas a OR A aon eee ae ae 1 p Ae PRE! De Po eg ee) (ees 8 | ee +2
ky + ae er e Seap ee reepere gem Pe 3 3) 2) Adobo ai ee +4
SD Wo 14. hos se tee PL. eee Se oe see eee 1) ABA ae ochre Beas + 6
4 Ger 18s 8. ooo So ohu cendked dbae meee lee geeiaeee 4 Ee See eres eee SE CPt +8
46 £0 18 ceo nec cnn ncaa nwcdn ne He -nblees|e be pie ate bes Pete ae late ane Geren Cea +10
TS GG rt a eR ee SC eee ena - ih Pegnd ae +12
DD GO DD. ood nicotinic an widie'bn on naide Se ou a sfonde] s cee erin ee epee tenon ai ee tne ae +14
DOGO DS oo cannons anne evmenccsa dunn eheh fete sl)s Dials esis SEs hehehe e ns emia a= +16
BPCQUEDCY. nc duo cas conn cep meee ics 23") 1S: fT LS Te ls See ae Sade ee
Departure from mean...2 ....5.2...-.-..-.- —2 |-1| 0 |4+1 |4+2 |+3 |+4 |+5 |+6 |47 [+8 |----.-
Table XIX shows how close a selection can be made for stalk
diameter in 2-year-old plants by saving only the tall plants. This
method, of course, is much quicker than to measure the diameter of
the shoots of every plant in the bed.
TaBLeE XIX.—Correlation between height (inches) and diameter of largest stalks
(45-inch units) in 1911 of greenhouse seedlings of 1910.
(Heights, subject; diameter, relative. Coefficient of correlation, 0.792+0.008.]
Diameter in 1911 (.4-inch units).
Fre-
Height in 1911. quen:
3/4/5|6|71/8 10 | 11 | 12] 13 | 14| © ;
Inches:
PRE a Rama tpey emia (ee ie Mame Meade Abele Hence Akon iba? faci Chan Pte res ee 1] <2
Chi se Bar seb betaine bie oufs ue Pert ie Beds) ks ea eee 0| —36
SEAT INSEE? HIRE RS Wed ARES BES Ae Se Re or a)
TERR OS tee eee, > BE Se FORE fee! led BST aid ied BR Sh SE 5| —30
EL SER OER ee a, MO Re Wy RR REGS PP GRR Kohn lads Me i Bes oe
AB Soues tee. faze us AP eR Fa RE FFs Nera ect | Re 7| —™
SRS, FRE UR EES SIE A, BTS besa thiheNe phe} t~scdoece| otake as 1] 4| 25 | 28 | 22 2.1°:9.ic ea 92} +9
OE, AREER GES EGR VERSES Saal yl? Ma ae 1 | 29 | 23 | 24 41... dcikhoounnl 83 | +412
Bi Pees RS Pe ae CARS ieee a Ra SY. 7) 9|14 6.11.1... age 42] 415
exes ee ca cae Si eas en EEL, , Viale dueBodedheadd 4|14 $1°s] 14a 2] +18
Ee RR SEES We SE o> cote RE A RE, OR NR 3/ 2 2 12} 421
OW SPREE ep ch a ad RR a Ga: GR Cee 2 $3. ...b0 Ce 6| +24
oars cc oaddudaW ences Cherie cule + ca] c> ocd em cab albandaladedlas «ab ibd ibeseeenste ne eae 1} +27
Bo ae Seek oes fac elec aa als « > le s kale coos ants Hace causa lke Ce dite Os SEA a 0| +30
TBS Soe ano che cideccuk sade dlveatss phe os-] «4 nolho, coh Giedanyelacaahd. cs Uiny is sean 0} +33
DR NAS itt Sta hae RR Mia SRG BR Bip oe MEA Ger ee te 1] +36
RPE eR SAE Fee) MN SERN PD ARE Rls KER PRS hare fey a Pye 0} +39
YR REO LE MES ER io} MARGE MEN SR Pm ays CDMA NEE ier me 1| +42
Frequency.........-.-. 19 | 64 [117 |171 J258 |132 |122 | 27 | 19 6] 0] 1] 949)
Departure from mean —3 |—2 |—1 | 0 /+1 [+2 |4+3 |+4 +5 +6 [+7 [+8 ]......
263
}
ns
:
| 3
a
-
4
b
;
oe
a
:
|
:
:
4
‘
BREEDING. 43
All these tables go to show that in asparagus we are dealing with a
stable plant with a permanent individuality ; that an individual char-
acteristic observed one year will persist in nearly unmodified form
in future years. Without a study of this kind any breeding work
would remain an uncertain proposition for several generations.
VIGOR OF SEEDLINGS OF MALE A7-83.
The several lots of hand-pollinated seed with the check lots of open-
fertilized seed which were sown in the greenhouses at Washington in
weer HEIGHT (VV INCHES
FEB. 11 19/0.
2
3
4
Ff
¢€
7
e
9
i)
“
f2
“as
“a
46
“Ss
‘7
1a
xs
@
2
SERN
Fic. 2.—Diagram showing the average height of 87 progeny rows of sbadthius of 1910 in greenhouse. The
measurements were made February 11, 1910, for comparison of open-fertilized and hand-pollinated
lots of seed. ;
January, 1910, were planted to study the effect of the different par-
ents on vigor of the young seedlings. These lots of seedlings varied
markedly in averagesize, and it was easily seen that the open-fertilized
lots of seed as a rule were shorter than the hand-pollinated lot
from the same female plant. The accompanying diagram (fig. 2)
shows the average heights on February 11 of the entire series of
seedlings arranged in classes showing the different lots of hand-
pollinated seed with the sample check from the same female. The
check is shown in black with the different pedigreed lots following
in outline. A study of the diagram shows that wherever male A7-83
263
44 BREEDING ASPARAGUS FOR RUST RESISTANCE.
was used an added vigor is shown in height over the check, which
represents an average of the selected males. Of course, this check
lot is influenced by the proximity of good or bad male plants, but
usually several males would be about equidistant from any select
female. Several other males show an added vigor, but the lack of
rust resistance shown in these lots when exposed to rust in the fall of
1910 removes them from consideration. ‘This difference of vigor is
still maintained in the seedlings of 1910 after growing two years in
the field. Of course, rust has entered into the effect now, but it cer-
tainly has not been the whole cause of the marked increase in the
progeny of male A7-83.
The size of seed being so important a factor in seedling size, it was
thought best to continue this study in 1911 on lots of seed of the same
20
Sie
\)
= /O
Wy On
‘ nvm, |
N 5
ee Eaahen= =
ee -— O- - --O~ ~
()
13 /4 15. ..S6 17 «18 AF 20a ae
CLAS S—HEIGHT 1N CENTWITE TERS
Fig. 3.—Diagram showing the height of 50 seedlings each from A7-25 pollinated with A7-19 (male) and
A7-83 (male), seed weighing between 0.021 and 0.024 gram.
average size. Female plant A7-25 was pollinated with both male
A7-83 and male A7-19 in the summer of 1910 and carefully weighed
seeds of the two lots were planted to show the effect both of seed
size and of vigor from the male, The effect of these two factors on
the seedlings when 10 days old is shown in Table XX. ‘To show the
effect of A7-83 on seedling vigor, 50 seedlings from each lot repre-
senting the different males were selected from seed weighing as near
as possible the same; in each case the weights ranged between 21 and
24 milligrams. In the diagram shown as figure 3 a comparison is made
between the heights 10 days after germination of lots of seedlings
from A7—25 pollinated with these two males. The result shows that
the added vigor of the lot from A7-83 is due to something besides
large seed.
263
ee Se ee ee eee
BREEDING. 45
TaBLe XX.—Comparison of weight of seed and height of seedlings at 10 days of age from
two lots of 1910 peste from A7-25 female, the two lots having different male parents,
_ A7-19 and A7-83.
Height of seedlings (centimeters). Weight fre-
quency.
Weight of seed. SE eet ee See PR a
12 | 13 | 14} 15] 16] 17]18 | 19 | 20} 21 | 22 | 23 | A7-19 | A7-83
ee A7-19| 1 1
13... ---.-----+-++-+--- FL ee NS rage Saat Rd as Sh NR ae VI Was BN aaa Bet Ro
‘ 7X21) Sn et ACRE. chee oh ihe cee ec eee esac
| ama 4 Aden LA bite Nis cae vet Chae Pa Poa ead Taba eters (ooo
15..----------- +--+ +2. Cf Hi es a eee Seale Ms I ie) Sak aa Sed Soe Bay Dee ed So
18 A7-19 ae) fC Sa aed A Pe re Pca ces
Gta BR SO, IOS © sd MN Fa Oa ORR I A Bae ah Geen a “Tae
a 17 A7-19 1 2 Oe ee ee ee 3 eeeecccs
oe eceeeerceeeeeeee eee 7 ey A SR NOS Se net AS Lee a RN Sete Bre eS Die ee es
% 5 em ed na ae i RE eRe A ead ee ee ee is Sitoe ee
er etceretese rere eeeee YES Sg GN RAY OS aS 5 I FRG TARR had Sed DERM AS Kiet oS 1
19 CORT ew eh bay tte ke. Bo aoe | Seen
eo reteeereteeeeeeeee- AT7-83 Re es Coe CEES cls APS Ca TR ER Od Be wie ct i
sis yo ee le WEIS 18 @ BPE bob i oe 5 Bae I
Bi Ua ww ween ee eee eee nese DN ed eee ae ies ercleh Ds nee SN RS AON TES SURI aE De 0
; _ RING es tee No. AN Ms SC Ge ey RE MaRS 3S Sere so? he Bein ts
BM) | Bbstenn---------------- FES SET ED re Se ihe Dedeosa lacs thes, che RN See 6
4 ou se a ae Rian eas bo aeh ee be Ses ee 17h es
: oO Tt oo Bae ee cee ri ey ae Pa a ie ee Py ee 8
a iy Oy eae Sea ad a Baia 4 och tes [toate re a inpeprgon
Bisco citer cn <2 = = >= => -55-- FAY OE Dg ee la ee i a Pr ¥ bodkes. oh eidtoses he 2
; be SS CG AE ig SE AMIR) sei Pea Ga i i eae Ae Sie Ce
Renae benny in> => = 5 s- == = - UGE ,Y SEN 2 7SeS Ree ry Son ae See eee tibet Pie aes 4
- A) all Clie alae a San Se 0 Sl Sa Mal 55 ER Biel pa cS see TS
TE a ea PAE DRG PERE Oe Rae rae MRR ces Sie Waa oom Baw es, Se iis He 2
ee 1a YOR INS PG RSET hae i THI ie, aaa Das ve ctaee is con SE catia! Pe eae.
aka ie) I RY ees heel NPG, a Sel NR MIR VE A SN ep Ua 1
Height frequency:
.. 2 Sa ae 1} 2] 6|/20}26/14/171) 7| 21.11....|.... rw Sapa fas.
ORE Na 2 OE he 217-4 56 bebe losses ee 97
RUST RESISTANCE.
To any one familiar with rusty asparagus fields the injury caused
by rust is apparent, yet the actual damage is hard to estimate on
account of the different seasons affecting the cut of the crop.
Table XXI shows the relation between the percentage of stem
cross section of the crop of 1911 to that of 1909 and the rust resist-
ance of 1910. Several plants growing in 1909 rusted so badly that
they died before 1911, and are therefore excluded from the table.
It is realized that the increase in size of asparagus hills is influenced
by many things other than rust, so that the actual effect of rust is
much higher than the coefficient given shows.
263
46 BREEDING ASPARAGUS FOR RUST RESISTANCE. — ;
~e agit
Taste XXI.—Correlation between the degree of rust resistance in 1910 and the pere reentage
of the stem cross section of 1911 to that of 1909 in row Al.
Rust resistance, subject ; percentage of stem cross section, relative. Coefficient of-correlatiae: camo)
Percentage of stem cross section of 1911 to that of 1909.
Rust resistance
in 1910.
20 | 60
BND Se cs —160]—120|—80]—40
In 1910 a study was made of the rust resistance of the seedlings
started in the greenhouse at Washington and later transferred to
a permanent place in the field at Concord. The rust attacked the
field in August, so that in the latter part of September some of the
nonresistant plants were dead. The seedlings of the several lots
were then judged as individuals, and Table XXII shows how they
ranked in rust resistance. Lots 1, 4, 8, 11, etc., are from open-fertil-
ized seed from the female plants used, while lots 2, 3, 5, 6, etc., are
from seed hand pollinated from male plants mentioned in the second
column of the table. A study of the table shows A7-83 to be pos-—
sessed of great power to transmit rust resistance. The results shown
in this table are those on which our breeding work is now based.
A7-35 is the only other male showing desirable resistance. This
male is being tested further and may be selected as a breeding
plant. But there is now no question as to the desirability of A7—
83 (Pl. X, figs. 1, 2, and 3, and Pl. XI).
TaBLeE XXII.—Rust resistance of individual greenhouse seedlings of 1910 in progeny
lots in the field, September, 1910.
Parent plants.
g Xo ) Ae WR a 4 15 6
SO RL ERS 1\.. Jal (41.6) 6.25
BIB we eke ee 2 ? | By Py Hm Pes Pam BS) age) ee eh MR, 8.50
BIRO 6 555 bcs eink 3\.. 4 ak Slo ee 5.47
EE SE. el Chetek babe ace 4\.. eleel Sakae Bee 5.45
AP-18. dec ae tes 5}. i Pee ee be 5.50
TV 5 We oo 6|.. ssahcol Alesis geal 7.39
APGba cheeses je 2} 1) 4). 1 5.30
AN Ava saitev manent aveds 8}. 1 1) 41.21 @.. 5.00
yO RC RC ay 9)... CORP aie wf 6.35
BTID. Tavendase ce 10}... 1% 21 21 6. 67
ROR ss, cachet een 11 1\..1 3] 1) 9 2... 5.74
y\ ees 12 ea | a 8.19
Ty a ee ra ee 13 23S 6.25
263
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE XI.
N&
: k~ > jhe
eae
PLANT “WASHINGTON,” A7-83, SHOWING THE GENERAL TYPE OF THE BEST BREED-
ING MALE USED IN THE RUST-RESISTANT BREEDING WORK.
(Photograph taken June, 1910.)
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE XIll.
PLANT ' MARTHA,” B32-39, SHOWING THE GENERAL TYPE OF THE BesT BREEDING
FEMALE USED IN THE RUST-RESISTANT BREEDING WORK.
(Photograph taken September, 1909.)
Taste XXII.—Rust resistance
Parent plants.
BREEDING. 47
of individual greenhouse seedlings of 1910 in progeny
lots in the field, September, 1910—Continued.
Individual rust resistance of seedlings (grades).
ber of | Mean.
‘
‘
St i
:
. tc sc
rk
Oo Hw: wrnn: e
Priel as WO te OL acne ee
Phases ie in ay aed al. = lsat ae 10] 5.70
Py Ne pee OG SSA a Ra a 1 | DD 15| 6.07
Giiee Mtb sls ocbe Bewtiad tleoote Aa SS ie 10| 2.70
na BSG oT OS | A a ee a Me 8 a Pe 8| 5.19
3, Sta} Ae Ta ee a 3}..] 1: fe A 7| 5.29
BRL eOCR Ie TE SEN Gia Lt+. Me: te i, Pd ed Be 18} 5.58
Peeks! Hue Sloot’ Bh Ur ab pt es A ri 10} 5.55
5) a Spr SS NU Sm PO ae etl Wa Cal HP 10} 2.00
Sh A RY A a ss le TS 10| 6.10
Peat FW ee Oe St Ole gl ald | es 19| 6.82
Repciat ath Hap al Et Sb OF Pah b 1 15| 6.23
Bay pS) 7) YR ah PSR Ve RG A PM a | 8| 9.50
BEG OP, 28 at Se A 3 SO ek 9| 6.39
Lame: See Phe Bh ato Pte be Peg ey pera 10| 5.40
Ss ee a a a By 3 Sy 9| 5.39
Sey SRY SE 05 a Se pee st Ae} MM Dye et Te 10| 4.90
ss fea ee ee I | | a as ee | | 10| 5.95
i Bete | OM ER OT TN Na | ae ee 11| 5.68
BE ea ay ee SE PE ald ee WRG ah Bll S48 EY 10| 4.20
Se ee ok ak Pah ede ie be 10} 3.60
der ees at ty OC Sp Tee Bh eens 10| 4.85
hee Ga eae AN | at amt se | ee Da yi Natl) St 8 9| 5.78
DPS A Te ate ke cle Shes PY Sb AD 4 Eee 10| 7.55
52 Loy OP abl Fon ER VN Re 1| 6] 2}. -}.c/. 9} 8.05
Be et eee or A Gy i | a any SO PO od a) Hae ils 10| 2.
BP OR Pa | | fe | ee BA ; 11| 3.
eelsch Dee Mak Pa BES oT Ae 10| 3.
MH Te A ae abe 10| 2.
a hao EE |) a Py ee a: a F a 10} 2.
on Be IPI Ry ‘ ede 4.
a a ts 2 in 8 3.
: = cal ae see a 4.
: en 2 x
Bek Se Fe Pa z a al. . i 10| 6.
mae ne | he vie 9| 5.
bike a 1 2 Q|..|. 20] 5.
Feta? ee Hie $2: Pat AB] fy: 14) 3.
ae, OE Se 3 1 i 19} 4.
| an a ; ; 5 10} 3.
ae 5.
4.
ce cos eo Oe
SSENSSSESRREAKSASHIASYSY
'
w
_
(=)
go on ge
bo
or
— || J | | | | | | | | SS | | | ES ef | | — | ————__—
48
Table XXIII shows the height of these greenhouse seedlings at a 3
end of the season’s growth in 1910. Again A7-83 is found standing
out above the other males in the transmission of vigor. As men-
tioned on page 44, in respect to the vigor of seedlings of some of the ©
males while in the greenhouse, some good lots are found, but they
are poor in rust resistance, and the male parents have been discarded.
BREEDING ASPARAGUS FOR RUST RESISTANCE.
TaBLeE XXIII.—Height of individual greenhouse seedlings of 1910, in progeny lots in
the field, September, 1910.
2
Parent plants. Height in inches. EI =
~ a
: ° =
°
a : F
Es | a
013|4 7|8| 9 |10/11/12 13/14
Ber ea 5|6)|8 As |=
J | ne eS 1\.. de 2| 3] 1/..1 3 -| 10} 12.20
‘A7-83| 21... sez ls ieee -| 14] 16.79
A7-85] 3}. | Bech ol ae ae 15| 16.00
0 2 | eee mee ee 4). fa ey a oY FG iia 10} 13.20
A7-19| 5}. . | 2} 2) a) alte. -| 10) 12.50
AT-3| Of. .\.|.012) Boo) tee ht 9] 14. 89
A7-85} 7|..|..| 1} 1]--| 1]--|--| 1] 2)--| 2}. 10) 12.10
YY eae ae | eas HV, Way td fese Eee eH | Ge: -| 10) 12.10
A423) 9). BN RRM UP 10) 15.10
A7-19 10). .|..| 1 ER aed Ped 08 35 ee 15| 16.00
ee ee av)... a EE ae be) 15. 64
A7-83)12). |. BY 6 ay a 9] 18. 44
A7-85|13}..|.. seheltctostaeloe (oa 18.10
Ce Seema ina}. |): 2[22!2.[ 2) a] a a] 14.
‘A7-19|15|..|_.|. 1)..| 1) 3] 1] 2; 3] 3! 12.07
B24-13/16)..|..|..) 1] 1) 2} 3 aj..| a)..| 1). 9.20
pe See Me ee Be 2 ae Ge ee 2)..| 1 1 8} 12.25
A7- 7/18)..|..|..|..|..| 21 1). -| 2)..|.. 10. 57
A7-19'19}..|..] 1]..| 3] 2} 7) 2] 2l..| a)__|. 8) 8.89
BI2- 1120): A. 2) Uo) Hd Ba 1 3 SSIs 10.00
B24-13/21]..| 1) 1] 2/..| 2} 1..| 2i..]} alt. 8.10
BSF-20). a | a My A HS RS, aR, a PR Dee) 15.10
A7-19}23)..)..|..|..|..]--[ 1 1]. 3] 1] 3 | 15. 21
A7-19|24)..|..|..}..]..|.-| 1) 2} 2} 2| 3} 2 5| 12.80
A7-83/25|..|..]..) 1]..) 2)..!.-1..] a 2) 1) 1 8} 11.63
W- 2i26..|..1..)..| a..}.-f..] alc! 21 2 14.22
BO-44).0 5... 27] 4)..)-2).-]..) 2)--l2-] 213) 2) 1 12.00
| Boee-s2iaah..)..|5-| 2) 2.) wes) al ale | 9.17
8 7 eS eee Po)... OO TG hea eG a ha ee 16.70
B12- 1/30|..}..|..|..|..|-.]..1.-[-.| 1} 1} 2} 14.90
Bas-63)/31|..]..|..]..}..].-) 1!..}.-]-.1 0) 2b 18.09
B90-10/32|..}..|..|..|..|..]..!..| 2| 2} 1. 15.00
B114-31/33)..j..|..|..]..|..| 1. 2] 1] a.-| i. 14.80
B84-44)...:..... 34) veh sh shod sd Alec ae 18. 20
AT -TOIBDIs «Wo <1 le los letdeclecl.cl ieelee 9) 19. 67
A7-19/36}... PS lca aa eas 20. 30
re || ry Miya AR SR A PPT Wa fg PB 20. 44
B98-32)38)..}..|../..] 2)..] 1'..|..]..| 2). 14.70
96-4430). .|.-}..J.]..|..]-./ 1/..|..|..1 Ll. 17.40
C©7-5-12)40)..}..). 2). 2] 2f2 202]. 7 Qt 2]02] 15. 60
C13-5-33/40}. foo] e221 a) 222) a. 9) 15.33
W= Wied. h os}. foci. ch...t..| al-gh ak 9} 16.78
Ba6-47).0510.4: Sy Ta 1). 4.3)... 3 15.70
Co a D7 1 | 14. 40
} W- 1/45). as BF, A Fn He i] 3) 2! 16.29
BS4-60!......... 79 Gk OE HD ak Dt 8} 11. 63
‘A7-83/47|..|..|..}..|..|2 3'..| aj. i 5] 12.07
Bgs-70/48}..|../..]..| 1] 3} 2, 2) 1/..|..] a. 9. 40
B92-43)......... 149)... | at ot ayal alt: | 12. 60
A7-19|50)..|..}..]..} 1] 1] 2; 3) 2) g/..].. } 11.07
B98-48)51)..|..|..]..]..}../../..] 1/..} 1] 2 15.00
B114-31)52}..|..]..| )../..| 1!.-}..] 2.1.2 | 13.50
geal Sse be lo). -]:-]-2)-.]--|--| | 2 i] 3} al 4). «|. -Jocleclefesfee} MM) 12.36
B98-32)54)../..}../..1../..]../ 1] 1) 3}..} ai [eee Fe
B132-26)......... ISB)=<|-<]c|aof2)--]--]-=|--|--] dl EETETS) 20) 16:90
A7-83/56}..|..|..|.-Joeleeloo|-ofoo].-foofe 3) il i] Sie 21.30
B&8-63'57!.. es 6 62 se 6's 1 1 ~ 1 Bs\ weectccton eetee eetes ee . 17.80
263
watlirat =.
ee ea a
BREEDING. 49
Taste XXIII.—Height of individual greenhouse seedlings of 1910, in progeny lots in
the field, September, 1910—Continued.
2
8
Parent plauts. Height in inches. rF s
3 S|
Z g| 4
a 1] g| 3
Q Xd || 3/45] 6] 7| 8] 9|10]11]12/13]14/15|16|17|18] 19|20|21|22}23/24)25 26|27|28)29'30/31|32 5 | S
ast: 4): 2... MEL Aa Meh obey a a Lt a (a es abt 9| 13.78
BGS. ohoale =|. [ote |- «fe =]. -fectest 2) LEE 38h .f. fl. Ais 10| 19.10
Ei 8 Te 1 PR a 1 a Ae ev vv B 2s 11| 12.36
7S A SS a a a STA Dt A a Ae 10| 12.10
B136-24)......... TR BV M8 eee? Oa A a a .¥ 10} 14.40
pear oh te. tale te tecke-t OP UT Be P aoc] ck] UY 2)-- |---| cl. ae 10| 15.30
Wi40-95). 25... “OE Sie SE Sa IN (ED We PS Oo a a 9 aS 10! 18.90
SO Re A we WD 1 a PM | a | a I)... 1| 10) 21.80
C13-5-33|66 J 8 as OS a a ie lH ek a el Gos 9| 14.56
B144-16)......... yA eS 8 a wR i YD ee ee ES 10 16.30
A7-19|68 Fg | a My ed a a a I cee 10| 14.10
C7-5-12\69 6 I a ee | a a 10) 12.50
C13-5-33|70 TH OR ee SN EC TT aa 0 ie i Se a ea ct 11| 11.18
C17-1-23)71 a 1 BW 5 9 PT eC 10! 10.20
- 272 Ey ON i pg ee) eae ee ne 2 2 ae a 10; 10.00
8-3-14) 22... Wee ee lec4 Uk 22] 2h.) See eee eee eek. 10, 10.20
A7-1974 SO) 208 eee TS BT Ae aah A a A 9, 9.33
C7-5-12175 OS OE DR rl BS AS SC SE a a Ye Wf a 10} 14.70
C6-3-31|_.......- 7° A Ra Gg) SS sah" GO a J A 9| 15.56
A7-83177]..|..|..|..|..|0.|-2|--}ec|--|-c1 1 1] 2) rie a | 10| 18.80
Se Se 1 A a a 10| 17.50
C13-5- 1]......... SS AR EY | A RD 9| 13. 22
A7-85 eR oc tol ie ef Saar ae a a) ol 20| 18. 80
a aR HR Vv 14) 14. 43
W- 1\82 WAL ts c|o) Hep ef 22) 2 Sf at a} a} 1) 2] a lL 19} 2036
ig 44) OOS Tae wy ee OO 10} 10.10
2% a Vv ee i 15| 12.47
7-85|85 1). -{.-/..| 3] 1] 1] 2] 5].-}..) 1] 2) a} aj ap 19| 13.11
C21-5-33|......... 86)... Bod Hole A BH B.-|- Pha ba 10} 13.90
2 0 ae a Ee et ee | De A
TR a a ee WR a | WY 1 | UY i Ce WT De RW I Dak
Total... =... 1) 5| 9112/2913) 58/50/75 /66)75)84'79)72 57/61/45/35,34/19124.11) 7) 3) 2] 2| 2).. 1) 1952) 14. 42
Among the females, B32-39 (Pl. XII) stands out in the 1910 test
as a good parent for rust resistance. Unfortunately the progeny lot
of seedlings B32-39 x A7-83 was in poor ground and did not show
well in vigor compared to the open-fertilized lot which was in normal
soil at the other end of the field. As was found in 1911, this apparent
lack of vigor was due to poor ground only.
The accompanying diagram (fig. 4) shows a comparison between
A7-83 and A7-19, the two males used most in our pollination work
in 1909, both in rust resistance and vigor transmission on the various
lots of progeny from different females. Accompanying the records
for each progeny is that of the check lot open-fertilized from the
same females. This table shows strikingly the great advantage in
using pedigreed seed of good parents. Attention is again called to
the fact that the male plants available for pollination of the open-
pollinated seed were, with few exceptions, select plants.
263
50 BREEDING ASPARAGUS FOR RUST RESISTANCE,
PERMANENCY OF RUST RESISTANCE. .
A study of rust resistance year by year shows the same permanence
of this character in the plants that was shown in the studies of size,
WERAGE RUST. RESISF-|
wine PF SLEDLINGS
2468
_ |AVERAGE erp ® OF SEEDLINGS
NCHL SF
40 45
SARENT PLANKS
9
A3=73
5.45 WOO
92. XAT-19| S /2. 5.50 <=
A4-73 wee
5.00 DROW
3.8 6.67
: WES
“ak 5.79 ee
A8&-9 an
5./9 LO
R SSE
B32-32 Peed
6.70 RXXsSSs
»» C re aca 6.82 7a
Leer ue ces LA pte ay ES “ 6.23
884-44
Mme re
F. ES ISO
5.78
7.$5
oa
3. TORS
5.07
BE er
4.35 SWS)
3.39
Ee
2.75 So
4.33
a
3.20 SSX
5,27
70
Iw cre
iy |
REE
==
eM ae
BSS SSIS
OPEN | TEVESES .9F OOO
Fia. 4,—Diagram showing the effect on greenhouse seedlings of 1910 of A7-19 and A7-83 with respect to
the average heights and average rust resistance of progeny lots in comparison with the progeny from
open-fertilized seed from the same female plants,
yield, etc. Once the individual plants are learned, their individuality
is recognized in different seasons. The attack of rust on the green-
house seedlings in 1910 was very uniform and satisfactory from the
263
|
|
|
BREEDING. 51
standpoint of selection of rust-resistant plants. In 1911 the rust
came on very much later and did not get started uniformly over the
plat. Some lots were attacked as badly as in 1910, but the ends and
outside rows where exposed to the wind and plants that were shaded
by trees failed to show as much rust asin 1910. In spite of this fact,
the correlation between the rust resistance for the two seasons of the
individuals included in the tests of both years is quite high (Table
XXIV).
TaBLE XXIV.—Correlation between rust resistance in 1910 and in 1911 of greenhouse
seedlings of 1910.
[Resistance in 1910, subject; resistance in 1911, relative. Coefficient of correlation 0.512+0.015.]
Rust resistance in 1911 (grades). Fre- | Depar-
Resistance in 1910. quen- Peas
0 1 2 3 4 5 6 7 8 9 | 10 | °Y- | mean
Grades
eee oe ray no pre ers sib tae ie Le oe Be Gees alee ac fase oe 1 1 sl ates fe 2 —5
LD kate eee 3 1 1 aes 1 2 te | Se ean 12 —4
yee dS 1 3 a 5 5 6 Pe 2 yA rR eR 36 =
i BE cepa) ee 1 2 7 5 13 13 34 34 LOM ae 119 —2
2s doc hint SSANE AAA eee Poe 3 5 i 12 47 38 23 138 —1
1 Ol CE EE enn Delete en on 1 2 1 15 22 40 74 40 13 | 208 0
erterese eee sleaet= 3s loos 2.5 - diay f sraeeet 3 5 13 32 60 39 22 175 +1
eee Pere ste close ale cc's eal tecSu Stswdeas 3 5 20 52 39 19} 1388 +2
ees. (eens stra nt a2 Po ee pe cate ale ates 1 9 19 29 26 84 +3
ec te e sys iL | Shee oi au metameeic wa heeicll ok ee 4 9 14 27 —4
ere eet oe (thats cli sot chee Geaapae was glticenet ts «teed it 3 2 6 +5
Frequency... 1 7 6 18 19 50 69 | 199 | 285] 192 99 | 945
Departure from
eS ee ee - —7); —6|] —5| —4] -—3] -—2] -1 Oop FE Po -pBleecee:
It should be noted that very few plants show greater rust in 1911
than in 1910. If the rust attacks had been of equal severity both
seasons, a much higher value for the coefficient of correlation would
have been obtained.
As was shown in Table XXI, there is a definite relation between
imcrease in size and rust resistance. The young seedlings in the
greenhouse lot show a relation between rust resistance and size,
but to a certain extent the size is dependent even yet on the size
of the seed. In Table XXV the heights in September, 1911, are
correlated with the rust resistance of plants as observed in Septem-
ber, 1910. The first 25 lots of the greenhouse seedlings were used in
this table, as the rust attack in 1911 on this row was more uniform
than on the other rows.
263
52 BREEDING ASPARAGUS FOR RUST RESISTANCE. _ a oo si
TABLE XXV.—Correlation between height in 1911 (inches) and rust resistance in 1910, we a3 ;
row 1 of greenhouse seedlings of 1910.
[Height in 1911, subject; rust resistance in 1910, relative. Coefficient of correlation 0.484+0.032.]
Rust resistance in 1910 (grades).
Height in 1911.
1 eR Se" SSSR. [ieee ee: Memes baRepege ese aes ae pI arse a a ee Es
BO epee ote eee wens ip Autsont i SPM : ie eee ee aces (PS
"ee a rerpien bet eee SEE 1 cp ae: Se
> Se ee. ere 1 BESS Ree yt ee Re Reeve Se
SR Ee ee aa pe yar 3 2 2 6 i ee eee PAE
oS Set a ae 1 5 5 7 § 2
«NES SNE eR Rte. SPO 2 2 2 g 5 8
5 Ss See eee 2 1 1 3 8 11 8
SE ARE Re 3 Le Pe oe 7 11 12
| SRC RE PERE AER Ces Prins, Danie 1 1 7 6 13
LASS Ee Onan Same nee fee ssa fee eee fered a yl SPY 1 5 9
ES EE RNS RES” Sot G us, Bee My! 1 3 2 3
ES ICIS EE a fT bP 2 4 1
rs eaels he Ook hee Doel Edo Penlhe ao Semel eae Cee ete ae een! 4
a nen Pobre bers rami frome = Fl. pS Nee a
ERP a eee tiers, nmin Pa de hot PRS aad 1
MD sacs een gba ess] stele ales oay Shee Maecenas
Frequency........- 5 12 14 16 54 60 61
Departure from mean...-| —5|—4]/—3]—2]|—1 Oj/+1}/+2/+3],+4
SEEDLINGS OF 1911.
In 1910 the work of making pedigree combinations was continued
in the spring months. This work was done before the rust developed
and was naturally of a more or less hit-or-miss character. Male plants
were selected for their individual qualities with the hope that they
would transmit these qualities to their offspring. A7-19 and A7-83
were used as check males to test new female plants. The female
plants that had given the best resistance both in 1908 and in 1909
were also used in making combinations with these males with the
plan that if any of the 1909 combinations showed desirable resistance
the 1910 lots of seed would furnish an additional supply of the desir-
able progeny.
The seed resulting from these pollinations was planted in 1911 at
Concord, and when the rust attack developed in August the behavior
of the different progenies was much the same as in 1910. A7-19
proved to be a failure in point of transmission of rust resistance and
has been discarded. A7-83, however, again performed in a very
satisfactory manner. Its progeny proved highly resistant to rust
and very vigorous in comparison with seedlings from American stock
lacking in rust resistance. Plate XIII shows a row of seedlings from
plants in row B24 (fig. 1) compared with the best resistant progeny
“Martha Washington” (fig. 2). The striking difference in the two
photographs is not so great as the contrast in the field, where the rich
green of “Martha Washington” contrasts with the gray brown of the
dead seedlings from row B24.
263
ig ae
os
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE XIll.
‘\
eo ig lve
Fia. 1.—WAKEMAN SEEDLING STOCK, SHOWING TOPS ENTIRELY KILLED BY RUST.
(Photograph taken September 25, 1911.)
Fia. 2.—" MARTHA WASHINGTON” STOCK (PROGENY B32-39 X A7-83), COMMERCIALLY
IMMUNE PLANTS OF STRONG VI@oR.
(Photograph taken September 25, 1911.)
PEDIGREE SEEDLINGS OF 1911 AFTER A SEVERE ATTACK OF RUST.
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture.
—
PLATE XIV.
oa
Ne
PEDIGREE SEEDLINGS OF 1911 AFTER A SEVERE ATTACK OF RUST, SHOWING VARIABLE RESISTANCE OF STANDARD GIANT ARGENTEUIL, ALL PLANTS
SUFFERING FROM RUST.
5, 1911.)
»)
-
(Photograph taken September
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture.
: aa, a. meme race.
PLATE XV.
‘
\
PEDIGREE SEEDLINGS OF 1911 AFTER A SEVERE ATTACK OF RUST, SHOWING PLANTS FROM STANDARD READING GIANT SEED, SOME NEARLY IMMUNE,
OTHERS RUSTY.
(Photograph taken September 25, 1911.)
PLATE XVI.
263, Bureau of Plant Industry, U. S. Dept. of Agriculture.
i
ul.
B
(LI61 ‘ez toqurieydag uayB} Ydvis0}0yd)
‘"JONVLSISSY SDVYHSAY
4O (€S-ZV) LNV1d JIWW34 V NO €8-ZY DNISSOUD 3O 103449 3HL ONIMOHS ‘LSNY 4O MOWLLY 3YBSASS V Y3LdV LLG] 40
SONITGS3SS 33uDIGad
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture.
PLATE XVII.
-FERTILIZED SEED OF B32-39, USUALLY
PEDIGREE SEEDLINGS OF 1911 AFTER A SEVERE ATTACK OF RUST, SHOWING PLANTS FROM OPEN
QUITE RESISTANT BUT LACKING IN VIGOR.
(Photograph taken September 25, 1911.)
|
Bul. 263, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE XVIII.
FiG. 1.—SEEDLINGS AT THE SOUTH END OF A BED AT CONCORD, MASS., IN AUGUST,
1910, JUST BEGINNING TO RusT.
Fic. 2.—SEEDLINGS AT THE NORTH END OF THE BED SHOWN IN FIGURE 1 ON THE
SAME Day, SHOWING THE DESTRUCTION OF PLANTS CAUSED BY THEIR PROXIMITY
TO A YOUNG BED ON WHICH CLUSTER CuPS DEVELOPED ABUNDANTLY.
EFFECT OF RUST ON ASPARAGUS SEEDLINGS.
BUD PROPAGATION. 53
Photographs do not show the dead color of plants injured by rust,
but in Plate XIV the effect of the 1911 rust attack is shown on
American-grown Argenteuil stock, and Plate XV shows American-
grown Reading Giant stock in an adjoining row. There is no ques-
tion that Reading Giant contains plants of greater resistance than
any to be found in any lot of Argenteuil grown on the station grounds
at Concord. Plates XIV and XV show this difference about as it
occurs in the regular field growth of the two stocks. The seedlings
from A7-83 progenies are superior to these standard strains in both rust
resistance and vigor (Pl. XVI). B32-39 gives very resistant seedlings
even when open fertilized with males of medium resistance (Pl. XVIT).
The small size of the seeds of this plant places the seedlings at a dis-
advantage, and the combination with A7-83 is needed to give vigor.
In the tests of 1911 no new progeny lots showed a resistance or vigor
comparable with that of ‘‘Martha Washington.” Most of the plants
tested will be discarded, a few females being held for a further test.
New selections from Reading Giant and from A7-83 progeny are
included in the pollination work in 1912. Some of these new selec-
tions show such a high individual resistance that it is practically cer-
tain that some of the new combinations with A7-83 will show great
resistance.
BUD PROPAGATION.
In order to increase the product of seed from individual plants it
will be necessary to carry out some methods of vegetative propaga-
tion. Preliminary experiments to this end were undertaken in the
ereenhouse in 1910. Seedlings planted January, 1910, were separated
when they had several shoots; the roots were divided more or less
evenly and the plants repotted. Nearly all of them grew, and in
about a month they were separated again. This was kept up until
in January, 1911, two seedling plants were represented by 60 or more
plants. When properly handled few plants die. In the fall of 1910
about half the crowns of No. 1 Washington, No. 2 Martha, B32—4, and
A2-23 were dug up, shipped to Washington, and planted in the green-
house. These crowns were split into several smaller clusters and
planted in 12-inch pots. New shoots started, but on account of low
temperature did not completely develop, and finally died back.
When the pots were moved into a warmer house the divided crowns
‘started growing again, and some of the plants have been divided a
second time. This method of vegetative propagation will be neces-
sary in breeding and seed-growing work.
j PEDIGREE.
In pedigree breeding work the performance of the parent indi-
viduals is not important in itself, and is only of value as it shows the
ability of the plant to transmit its good qualities to its offspring.
263
4
54 BREEDING ASPARAGUS FOR RUST RESISTANCE,
These plants selected for breeding purposes become valuable only as
their progeny show uniformly high quality and yield. So when a
pedigreed progeny shows a high commercial value its parent plants
become of great importance. They should be increased as fast as
possible by clonal propagation and should be isolated and allowed
to produce as much seed as possible. It is now that records and
history become important. Careful record should be kept of their
original source, etc., and future development.
In carrying out the work on this breeding project and of private
breeding work developing from it the following scheme will be used:
Number.—Each plant that proves of value as a breeding parent
will be assigned a permanent serial number. These numbers will not
be given to a plant until its progeny show it to be of value as a breed-
ing parent. Its preliminary records will be kept under a temporary
number used to mark its location in the testing plat.
Name.—Plants used to produce progenies for commercial planting
will be given names as follows: The male plants will be given sur-
names as Washington, Wilson, Prescott, Wheeler, Moore, etc., the
name assigned to one plant not to be duplicated in the future.
Female plants will be named by assigning them different feminine
names, as Martha, Mary, Edith, ete.
Progenies will take their commercial or trade names from the two
parents. Thus the progeny of No. 2 Martha X No. 1 Washington
will be known to the trade and growers as ‘‘Martha Washington”’;
No. 3 Edith x No. 1 Washington would be ‘‘Edith Washington”;
No. 2 Martha X No. 4 Wheeler would be ‘‘Martha Wheeler.” In
this way each progeny would by its name indicate its parents.
Records.—In keeping pedigree records the loose-leaf record book
will be used. A primary sheet for each parent admitted to registry
will be used, giving its history, description, etc. The performance of
the plants as shown by their progeny records will be filed under the
female parent as secondary sheets. An abbreviated record of these :
progeny sheets will be filed under the male parents as secondary
sheets to show the performance. .
No. 1. Washington ¢.
Pedigree: Q unknown.
$ unknown. ;
History: Original plant found in 1908, location A7-83. New American Concord-
grown stock by Anson Wheeler. Marked as best male in type and rust
resistance. Used in 1909 and 1910 in crossing work. In 1911 used as test
male in all crossing work.
Progeny: Very resistant to rust and showing an added vigor above open-
fertilized progeny no matter what female parent was used.
Propagation: Part of original parent dug up in 1910 for clonal propagation.
263
:
PLANS FOR DISTRIBUTION. | 55
No. 2. Martha 9.
Pedigree: 9 unknown.
g¢ unknown. .
History: Original plant found in 1908, location B32-39. Reading Giant stock.
Marked as best in rust resistance 1909; rather small type; used in 1909 and
1910 in crossing work. In 1911, under cage, crossed with No. 1.
Progeny: Open-fertilized lots of 1909 and 1910 better in resistance than any
other open-fertilized lots tested. X No.1 progeny best for resistance and
type of any seedlings grown.
Propagation: Part of original parent dug up in 1910 for clonal propagation.
When plants from any named progeny develop as good breeding
parents they will be assigned new names and handled as distinct
parents, their history and pedigree being recorded on their original
pedigree sheets.
When by vegetative propagation the original parent plants have
increased so that different growers have lots of the same progeny, in
offering them for sale the grower’s name should accompany the
progeny name for purposes of identification in case any error creeps
in; as, Martha Washington (Frank Wheeler stock), Martha Washing-
ton (C. W. Prescott stock). The registry of new parents for breeding
purposes should be through a central breeding organization, so that
no duplication of names will occur. For the present this work can
be done at the experimental station at Concord. These new progeny
lots must be tested in competition with some standard progeny of
known rust resistance and quality and their general value determined.
PLANS FOR DISTRIBUTION. -
When sufficient stock of any progeny is obtained to warrant dis-
tribution to interested growers, plans will be made to plant the stock
under conditions favorable to the satisfactory testing of these prog-
enies for resistance to rust. The lots of seed or seedlings issued by
the Department will, as far as possible, be sent to growers who will
be in a position to aid in extending the cultivation of the rust-resistant
strains.
SUGGESTIONS TO BREEDERS AND GROWERS.
In giving advice in regard to asparagus breeding at this time it
must be remembered that our experiments are only just begun.
Later results are liable to change the methods of procedure to be
recommended, but the methods and practices at present followed are
here outlined.
RUST RESISTANCE.
If rust is a factor in the region where the work is to be done, resist-
ant varieties are of prime importance. In order to secure resistant
selections rust must be present in abundance. Unless one can pick
263
~
56 ' BREEDING ASPARAGUS FOR RUST RESISTANCE.
the one superior plant out of a thousand in point of rust resistance
the work will be hard.
Late fall is the best time for making field selections, because at
that season the rust will be developed sufficiently to have marked
the nonresistant plants in the field so that they can be disregarded.
In providing rust for this work in New England it will usually be
sufficient to leave an area of nonresistant plants in one corner of the
field, preferably that from which the prevailing winds come. If
there was plenty of black rust the preceding season, the spring stage
will develop in sufficient abundance to provide rust for infection _
work later in the season. A bed of young asparagus not ready to
cut for market is usually sufficient to provide a lodging place for the
spring rust. Artificial inoculation has not been necessary at any
time in our breeding fields. :
ISOLATION.
After two mated plants have had their progeny tested and have
proved their value as a breeding pair they will be dug up and propa-
gated by crown division to secure a stock for breeding. This stock
will be isolated and used only to grow seed.
Isolation will be secured by building an insect-proof cage over the
field or by planting remote from other fields or wild plants, so that
bees will not be able to carry in foreign pollen. The mesh of any
cage will have to be small enough to keep out the small wild bees.
One of the probable methods will be to grow the plants in the green-
houses in the winter. During the winter of 1910-11 im Washington
we have been very successful in setting seed in the greenhouse by
hand pollination. In making seed plantations a grower will not be
limited to one female plant—any number may be planted with one
male. Whenever it is desired to use two males a separate field must
be used for each.
PROGENY BED.
In planting seed for a progeny test a uniform piece of good land
is necessary. The presence of shade, such as overhanging trees,
near-by buildings, etc., should be avoided. The bed should be set
so as to be uniformly exposed to the attacks of rust from near-by
infection plats. Any marked difference in moisture supply is apt to
interfere with the test.
As it is not the intention in the progeny test to grow large plants,
the custom at the Concord station has been to plant rather late in
May so that all danger of frost and also of the first crop of beetles is
past. About 10 feet of seedling row is sufficient for a fair test. Of
course, many lots of seed will not plant so much as that, but it is a
263
SUGGESTIONS TO BREEDERS AND GROWERS. 57
useless waste of space to take any more. Rows are first laid out
with a line and then made about 2 inches deep with a hoe. The
seed is sown by hand and covered with a rake. Skill in planting is
acquired by experience, the intention being to drop about six seeds
to the foot. A space of 18 inches between rows is ample to allow for
passage and cultivation. The two things to judge in the first year
are height and rust. The rust on the seedlings is closely correlated .
with the rust of the plant in future years and height is correlated
with size and vigor. The first year progeny test will eventually be
the main test of any plant’s value in breeding work.
* The use of a standard or uniform lot of seedlings as a check on
rust infection is desirable, and where accurate results are expected
is necessary. In our work up to 1912 we have used Reading Giant.
Pedigree stock of good quality alternating with rusty stock will be
planted hereafter as a double check.
VALUE OF BREEDING METHODS.
If asparagus growers ever hope to secure reasonably uniform
strains of fixed type, the methods of commercial seed production
will have to be changed from their present unscientific condition.
With few exceptions no attention is now paid to the male parent and
little effort is made to get good female plants, the process of seed
selection consisting largely of going into a field that has made a good
growth and harvesting seed stalks that have well-grown seed.
Mr. Frank Wheeler, of Concord, Mass., has for several years made a
practice of selecting the best male and female plants in regard to
type, vigor, yield, and rust resistance. These plants have been
allowed to grow and bloom during the cutting season. The seed is
saved from only those stems of the female plants that bloomed
before the general field plants came into flower. These seed plants
are the progeny of imported Argenteuil stock and produce a very
desirable quality of seedlings.
In the spring of 1908 about 400 one-year plants of this strain were
planted in comparison with a similar plat of a strain known locally
as “Small”? Argenteuil. The yields from these two plats were kept
in 1910 and 1911, as shown in Table IV on page 26.
This difference in yield is apparently due to the difference in the
strains in which the selection for large stalks by Mr. Wheeler has
been an important factor. No apparent difference was noticed in
the comparative rust resistance of the two lots, so that rust does not
enter as a factor.
If the above striking difference exists iijough the simple selection
methods used by Mr. Wheeler, would not other good farmers be
263
58 BREEDING ASPARAGUS FOR RUST RESISTANCE,
justified in trying pedigree methods in growing seed? The above-
mentioned strain is not pedigreed from either side, the parentage
complex including about 20 individuals of each sex. Mr. Wheeler
-in 1910 and 1911 planted his lots of seed from each female in separate
rows. The difference was so striking that in the future pedigree
methods will receive more attention.
PROTECTION FROM BEETLEsS.!
One thing to be considered in seed production is the effect of the
red or twelve-spotted asparagus beetle (Crioceris 12-punctatus), the
larval stages of which live in and destroy the asparagus berries.
This beetle proved a serious factor in the breeding work last year,
and is liable to become worse as time goes on. The first specimens
of this beetle found in Concord were discovered in the fall of 1908.
The fall of 1910 showed nearly as many as of the ordinary species,
Crioceris asparagi. Paper bags are not sufficient protection, as in
several cases the berries under bags were destroyed. The beetles
had either laid their eggs before the plants were bagged or else
crawled up inside through the open spaces around the stems. Cages
of 16-mesh wire fly screen keep out the red beetle but let in the
smaller specimens of the common asparagus beetle. Both kinds
may be kept out by the use of 18-mesh wire screen, which will be
hereafter used.
PROTECTION OF NONIMMUNE FIELDS.
Spraying methods have been developed by different experiment-
station workers in the past that if carefully followed by the grower will
keep down the rust. The trouble in applying sprays and the high cost
of their efficient application has kept many good growers from using
them. Some farmers have gone out of the asparagus business while
others have secured the best stock they could find and by careful
methods have kept on. The high prices caused by increasing demand
and lessening supply has made the profit in asparagus really higher
than it was before the rust became known in the country.
It is now certain that by proper pedigree breeding work the whole
question of noticeable rust injury in asparagus may be eliminated.
At the same time the pedigree breeding work will make uniform and
vigorous strains, thus greatly increasing the yield per acre. The
elimination of rust as a factor in asparagus growing will render larger
yields possible, so that the market price in many locations where rust
now prevents adequate returns will fall within reach of the large body
of consumers. At present in most regions asparagus is a luxury.
1 For a full discussion of the two asparagus beetles and of the methods to be used for their control, the
reader is referred to Circular 102 of the Bureau of Entomology, U. S. Department of Agriculture.
263
“
y
bg ?
M
—— >. ee ae
SUGGESTIONS TO BREEDERS AND GROWERS. 59
SUGGESTIONS FOR RUST PREVENTION.
Although the breeding work being carried on with asparagus will
eventually lead to the control of rust in commercial plantings, several
years must elapse before this result will become effective. Meanwhile,
it is necessary to take all measures practicable to prevent the destruc-
tion of existing fields of asparagus by therust. To this end the main
factor is to keep the rust away from the fields in summer just as long
as possible.
As pointed out by Smith and others, wild asparagus growing around
the borders of the fields, along fences, ditches, etc., is one of the worst
enemies of the grower. These wild plants act as infection centers and
their influence can be easily traced later in the season when the cutting
beds have grown up. During the summer of 1910 the writer made an
examination of the fields near Concord just at the time the rust was
coming on and in every case of infection was able to trace the cause to
asparagus plants that had not been cut up to the close of the infection
period of the spring rust (Pl. XVIII). When rust was found in a
commercial field by following it up to the northwest, the direction
from which the prevailing winds come, a young bed, an old neglected
bed, or wild asparagus was found in every case and always with the re-
mains of cluster-cup infections. Wild plants wherever found should
be dug up and burned. New beds should be planted only at rare
intervals of time and then if possible where they will be to windward
of a cutting bed. Keep the seedlings out of the cutting bed, at least
let none stay in at the time the bed is allowed to grow up after the cut-
ting season. Allow no poor shoots to grow up in the cutting field.
In other words, keep down every shoot of asparagus until the middle
of June in the latitude of Boston and see that neighboring farmers do
the same. In the fall the tops should be removed carefully from
1-year-old beds that are not to be cut the next year. This will in a
large measure reduce the liability of infection from this source.
The writer does not recommend the removal of tops from a mature
bed in the fall. The ordinary practice in the vicinity of Concord is to
leave the bed undisturbed in the fall so that the tops will act as a
winter cover and prevent the blowing of soil or snow. In the spring
these tops are cut with a disk harrow. Fields in which this treatment
had been used have been examined for spring rust after the bed had
grown up at the end of the cutting season, but in no case have cluster
cups been found. The Massachusetts station has at Concord a 3-acre
fertilizer experimental plat on which plants have been infected during
1909, 1910, and 1911 from young beds near by that were not being
cut. No cluster cups were found in this 3-acre bed except on plants
left for breeding purposes. :
263
60 BREEDING ASPARAGUS FOR RUST RESISTANCE.
SUMMARY.
Puccinia asparagi, the European asparagus rust, was discovered in
America in 1896 and in the next six years spread over the asparagus-
growing regions of the United States, causing great damage. In the
Eastern States no successful remedy was found, although some strains
were found to be more resistant than others. Among the resistant
varieties were Argenteuil and Palmetto.
The Massachusetts Asparagus Growers’ Association, organized in
1906 to obtain a resistant variety by breeding, secured the coopera-
tion of the Massachusetts Agricultural Experiment Station and the
United States Department of Agriculture in establishing experimental
grounds for this work at Concord.
Previous work on the life history of the disease shows that the rust
in all its stages occurs only on asparagus and that the uredo stage is
the most injurious. The injury is due to the mechanical and physio-
logical effect on the summer growth which prevents the storage of
food supplies for the growth during the next cutting season.
A large number of strains from America and Europe have been col-
lected andtested forrust resistance. Novariety proving uniform orsat-
isfactory, breeding work was undertaken to produce a stock that would
be commercially immune. Some wild species have been imported from
the Old World and one or more hybrids have been produced.
In making selections for rust resistance several acres of the best
stock obtainable were used. From the different strains several
hundred plants have been selected for pedigree testing after being
subjected to attacks of rust.
Rust resistance in asparagus seems to be based upon structural
differences. Vigor is not necessarily correlated with resistance.
Breeding work in asparagus is complicated by the fact that the
species is dicecious, so that two parents must always be used in seed
production. Hand pollination is used for pedigree work.
Progeny tests of select plants have been made each season since 1909.
The rust resistance and vigor of these seedlings have determined the
value of the breeding parents. The test male A7—83 and the test fe-
male B32-39 have given a very superior progeny, which has proved
satisfactory as a ‘‘commercially immune” type. This progency has
been named and plans are under way for its production in quantity.
In carrying out the breeding work, studies have been made of the
effect of the weight of seed on seedling vigor, the effect of seedling size
on the plant in the field, ete. Correlations between size of plant, yield,
rust resistance, etc., have been of value in carrying out the work. .
Bud propagation of select breeding parents has been inaugurated
to promote more extensive seed production.
Breeders and growers are advised to take up pedigree breeding to
produce good strains and to use careful methods in keeping rust out
of producing fields.
263
O
»
ase Oe eee ee a eee
Issued January 8, 1913.
U.S. DEPARTMENT -OF AGRICULTURE.
BUREAU OF PLANT INDUSTRY—BULLETIN NO. 264.
B. T. GALLOWAY, Chief of Bureau. |
THE PURPLING CHROMOGEN OF A
HAWAILAN DIOSCOREA.
BY
HARLEY HARRIS BARTLETT.
Chemical Biologist, Drug-Plant, Poisonous-Plant, Physiological,
and Fermentation Investigations.
WASHINGTON:
GOVERNMENT PRINTING OFFIOE.
1913.
BUREAU OF PLANT INDUSTRY.
Chief of Bureau, BEVERLY T. GALLOWAY.
Assistant Chief of Bureau, WILLIAM A, TAYLOR,
Editor, J. E. ROCKWELL.
Chief Clerk, JAMES E. JONES.
DRUG-PLANT, POISONOUS-PLANT, PHYSIOLOGICAL, AND FERMENTATION INVESTIGATIONS.
SCIENTIFIC STAFF. q
Rodney H. True, Physiologist in Charge. \
A. B. Clawson, Heinrich Hasselbring, C. Dwight Marsh, W. W. Stockberger, and Walter
Van Fleet Physiologists.
Carl L. Alsberg, H. H. Bartlett, Otis F. Black, H. H. Bunzel, Frank Rabak, and A. F.
Sievers, Chemical Biologists. . ;
W. W. Eggleston, Assistant Botanist.
Lon A. Hawkins, 8. C. Hood, G. F. Mitchell, James Thompson, and T. B. Young, Scientific
Assistants.
Hadleigh Marsh, Assistant.
G. A. Russell, Special Agent.
264
2
LETTER OF TRANSMITTAL.
U. S. DerarrMent or AGRICULTURE,
Bureau or Puantr INpustry,
OFFICE OF THE CHIEF,
Washington, D. C., September 10, 1912.
~ Sir: I have the honor to transmit herewith and to recommend for
publication as Bulletin No. 264 of the series of this Bureau a manu-
script by Mr. H. H. Bartlett, Chemical Biologist, entitled “ The
-Purpling Chromogen of a Hawaiian Dioscorea.” This paper, sub-
mitted by Dr. R. H. True, Physiologist in Charge of Drug-Plant,
Poisonous-Plant, Physiological, and Fermentation Investigations,
deals with a subject of great interest to both physiologists and ge-
neticists—to the former because of the important function in the oxi-
dizing mechanism of the plant cell, which has recently been ascribed
to the vaguely characterized plant chromogens; to the latter because
of an increasing need for the chemical identification of the “ unit
characters” for color in plants.
Respectfully,
B. T. Gattoway,
Chief of Bureau.
Hon. James Witson,
Secretary of Agriculture.
264 |
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folline jamefeisl (heed doles TE Le
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PALIN oo ee ke
Reerence oF Lhe cnrumomen. oa
inant Lie Chromogen. 2. 2 te ee ee
. | Peeicat and Chemical yproperties x26 01202 leas tae es ot
meenemce-of a second, chromogen... S22. ea et
Nomenclature_-_--------~----------------~---------------------
Possible relationship with the anthocyanins_________-_---+____~
Wheldale’s theory of anthocyanin formation___________--______
the Aa lta a ee a ie at ah Seep a ei 5 2 A Rb _a
T rn r
ILLUSTRATIONS.
PLATE,
PLATE I. Aerial tubers of the Hawaiian bitter yam___-_--_----~-
TEXT FIGURE.
264
Fig. 1. Leaf of Dioscorea, showing petiole enlarged at base and ence im-
mature aerial tuber, and bases of racemes______-_____-__
Page.
ste bern oon a t tas at apes catsghid iy ba
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B. P. 1.—76.
THE PURPLING CHROMOGEN OF A
HAWAIIAN DIOSCOREA.
INTRODUCTION.
In 1903 the Office of Foreign Seed and Plant Introduction of the
Bureau of Plant Industry received through Mr. Jared G. Smith,
Special Agent of the Department of Agriculture in Charge of the
Hawaii Agricultural Experiment Station, specimens of a Dioscorea
under the native name “ Hoi.”! These were inventoried as Nos.
10311 and 10312? and cultivated at the subtropical gardens at Miami
and Gotha, Fla. Because of the confused state of the genus it is not
now possible to assign a specific name to this Dioscorea.* Perhaps
the taxonomy of the group to which it belongs may be treated in a
future paper. For the present our plant may be conveniently re-
ferred to as the Hawaiian bitter yam. It has recently been offered
to the horticultural trade by Childs * under the name of “ air potato,
or giant yam vine.”
The name “air potato” has reference to large aerial tubers which
develop in the axils of the leaves. Many species of Dioscorea have
these tubers, but none exhibits them to greater perfection than the
Hawaiian bitter yam. The tubers (shown in natural size in Pl. I)
are propagative organs. In this country they are the sole means of
reproducing the plant, for all the tubers originally imported seem
to have been obtained from pistillate vines. However, the writer is
informed by Dr. E. V. Wilcox, Special Agent in Charge of the
Hawaii Agricultural Experiment Station at Honolulu, that in
Hawaii both pistillate and staminate vines are found. Many yams
1The name “ Hoi” seems to be of generic application to yams in Polynesia. Hille-
brand, commenting on a species which he describes under the name Dioscorea sativa, says:
“The yam, common in-the forests of the lower zone, was cultivated for the supply of
ships before the introduction of the potato, particularly on Kauai and Niihau. The
species ranges westward over all the regions lying between the Hawaiian Islands and
- Africa, and its native name, ‘ Hoi,’ follows it to Sumatra. The axillary buds are called
‘alaala.’ ’’—-Flora of the Hawaiian Islands, p. 438.
2 Bulletin 97, Bureau of Plant Industry, entitled “ Seeds and Plants Imported during
the Period from December, 1903, to December, 1905,”’ p. 9. 1907.
2In Bulletin 97, Bureau of Plant Industry (loc. cit.), the name Dioscorea divaricata
was used with a question mark. From this species, however, the plant described in this
paper differs in many characters, of which the most striking is the radial rather than
dorso-ventral disposition of the tissues of the aerial tuber.
4Childs’s Combination Catalogue for 1910, p. 3 of cover.
264
“1
8 THE PURPLING CHROMOGEN OF A HAWAIIAN DIOSCOREA.
which, so far as known, have only one sex are propagated exclusively
by vegetative means.
There is a chromogen in the aerial tubers of the Hawaiian bitter
vam which is of unusual interest in that it appears to be related to
the class of ammonia-greening anthocyanins. It has been made the
subject of considerable study in the hope that it might afford a clue
to the better understanding of these compounds.
OCCURRENCE OF THE CHROMOGEN.
The freshly cut surface of an “air potato” is greenish white, but
on exposure to the air it quickly becomes brown by the oxidation of
a chromogen (probably a tannin) which has not been investigated.
If the freshly cut surface is treated with ammonia, the natural green-
ish color is greatly intensified, especially in the region of the cambium
immediately beneath the cortex. Occasional tubers have pale-
purplish instead of greenish flesh, and many greenish tubers become
purplish in part when sprouting. If treated with ammonia these
purplish tubers also color, just as the greenish ones do, to a some-
what intensified green. On the contrary, treatment with acids will
not change the color of a greenish tuber to purplish. If, however,
the juice is expressed from a greenish tuber or from the green tips
of growing stems and acidified with acetic or hydrochloric acid, a
pinkish or purplish color of considerable intensity develops. Never-
theless, the intensity of the green which resulted when a cut tuber
was treated with alkali seemed to be out of proportion to the pink
which resulted from acidifying the juice. Therefore, in addition to
the pigment which was obviously one of the numerous compounds
classed as ammonia-greening anthocyanins, a second substance was
sought, which like the anthocyanin would give a green reaction with
alkali, but unlike the anthocyanin would be colorless or nearly so in
acid solution. After much experimenting, such a compound was
isolated—a chromogen which in dilute neutral or acid solution is
yellowish, which forms intensely wine-red solutions on oxidation
with plant oxidases or weak inorganic oxidants and intensely green
alkali salts both before and after oxidation.
ISOLATION OF THE CHROMOGEN.
As the result of several trials the following method is recommended
for the isolation of the chromogen in a comparatively pure state.
iIn regard to the anomalous morphology of the Dioscorea tubers, see the following:
3ucherer, Emil. Beitriige zur Morphologie und Anatomie der Dioscoreaceen. Bib-
liotheca Botanica, no, 16, 1889.
Dale, Elizabeth. On the Origin, Development, and Morphological Nature of the Aerial.
Tabers in Dioscorea Sativa. Annals of Botany, vol. 15, 1901, pp. 491-501.
Goebel, K. Die Knollen der Dioscoreen und die Wurzeltriiger der Selaginellen, Organe,
welche zwischen Wurzeln und Sprossen stehen. Flora, vol. 95, 1905, pp. 167-212.
264
Te.
a
Bul. 264, Bureau of Plant Industry, U. S. Dept. of Agriculture.
AERIAL TUBERS OF THE HAWAIIAN BITTER YAM.
(Natural size. )
PLATE lI.
PHYSICAL AND CHEMICAL PROPERTIES. 9
The tubers are sliced into narrow strips and dried as expeditiously
as possible at 60° C., or if plenty of material is available the tubers
are pared and the parings only are dried. The dry material after
having been ground is extracted at the room temperature, first with
ether and then with a mixture of equal volumes of ether and alcohol.
To the ether extract a small quantity of alcohol is added and the
ether distilled off, leaving the chromogen in alcoholic solution. This
is mixed with petroleum ether, and water is added to bring about a
separation of a petroleum-ether layer (containing wax, etc.) from
the alcoholic layer (containing the chromogen). The latter is added
to the ether-alcohol extract.
The ether-alcohol extract is distilled until the residue is free from
ether and considerably concentrated. It is then mixed with a cold,
concentrated calcium-chlorid solution (or mixed with water and
saturated with common salt) and shaken iirst with several portions of
petroleum ether to remove various impurities, and then with acetic
ether to remove the chromogen.’ The chromogen in acetic-ether
solution is then shaken with a large excess of concentrated aqueous
lead-acetate solution. The copious gummy precipitate, most of which
remains suspended in the acetic-ether layer, is filtered off on a Buch-
ner funnel and washed with acetic ether. The acetic-ether filtrate,
which is of a clear golden-yellow color, is now shaken successively
with solutions of ferrous sulphate (strictly free from ferric salts and
dissolved in recently boiled water) and bipotassium hydrogen phos-
phate and finally several times with distilled water. After filtering
the solution it is poured into a large excess of petroleum ether. The
chromogen is thrown down as a finely divided, white, opalescent pre-
cipitate, which speedily coalesces to form a soft, brown, resinous
mass. In this condition the chromogen contains much acetic ether.
It may be partially dried in a vacuum desiccator, then pulverized
as much as possible, and dried at 78° C. in a vacuum drying oven in
a slow stream of rarefied hydrogen.
PHYSICAL AND CHEMICAL PROPERTIES.
The chromogen as obtained by the process outlined is a brown
resinous compound soluble in alcohol, acétic ether, and chloroform,
moderately soluble in ether, and insoluble in petroleum ether and
water. Its melting point is not sharp, and it has not been possible
1 The saline alcohol and water solution which remains after this operation soon changes
from yellow, which color is imparted to it by a small residue of unextracted chromogen, to
a fine purple. The purple compound is the same oxidation product of the chromogen
which is mentioned on page 10 of this bulletin. Its formation in the presence of chlovids
is no doubt analogous to the bluing of guaiac resin by chlorids, as described by Alsberg.
(See Beitrage zur Kenntnis der Guajak-Reaktion, Schmiedeberg-Festschrift, 1908.
Supplement, Archiv fiir Experimentelle Pathologie und Pharmakologie, pp. 39-53.) The
oxidation product is precipitated by lead acetate, so that any small quantity which may
contaminate the chromogen after salting out is removed in the next operation.
62097 °—Bull. 264—13——2
10 THE PURPLING CHROMOGEN OF A HAWAIIAN DIOSCOREA.
to obtain it in a crystalline condition. It is fairly stable in neutral
solutions, and in alcoholic solution is unchanged by boiling with zine
dust under a reflux condenser. Prolonged heating in contact with the
air, more particularly when a reducing substance, such as zinc, is not
present, results in its destruction by oxidation. Acid solutions, which
are of the same color as neutral solutions, slowly deposit insoluble
brown compounds, with simultaneous destruction of the chromogen.
The chromogen is the first member of a series of acids. The other
members are successive oxidation products of the chromogen. Each
acid forms a series of salts. The relationship of these derivative
compounds is as follows:
by oxidation with peat = red brown
{ Yellow oxidation [PY further oxidation) |. idation
chromogen (¢———-——_—_—__——_——) product of product of
by reduction with Zn alle as
4 - ada
S| | ol lz =||2
+1] +1 |o 7 &
454 = brown
ammonium
ammonium salt of
green ‘. Phe reen ee
F Ey spontaneous oxidation|ammonium]by further oxidation
aoa. (oo eee brown ?
3 oxidation oxidation
product product of
chromogen
chromogen
The oxidation of the chromogen to its red derivative and the
formation of the intensely green ammonium salt of either the chromo-
gen or its red derivative constitute the two characteristic color reac-
tions which are mentioned on page 8.
The red oxidation product is best observed when the chromogen is
dissolved in acetic ether and shaken with water containing a trace of
fuming nitric acid or ferric chlorid. Under these conditions a beau-
tiful wine-red acetic-ether solution of the oxidation product is ob-
tained. If it is carefully washed by repeatedly shaking with distilled
water the red solution may be preserved for several days, but not
indefinitely, for oxidation will spontaneously proceed until the solu-
tion becomes brown by the formation of a further oxidation product.
Even this brown compound does not represent the final stage of the
oxidation of the chromogen, as a further change is indicated by the
ultimate deposition from solution of a substance insoluble in acetic
ether. The red stage in the oxidation of the chromogen is very easily
overstepped, with the immediate formation of brown compounds, if
the mineral oxidants used are too concentrated or if the chromogen is
not protected from a too violent oxidation by being dissolved in a
solvent, such as acetic ether or chloroform, which is immiscible with
the solvent of the oxidant (water).
The chromogen may be oxidized, in aqueous solutions containing
sufficient aleohol to prevent precipitation, through the agency of plant
oxidases (see p. 12) or of halogen salts (see p. 9). Under such cir-
264
a ie Sel ‘gy
PHYSICAL AND CHEMICAL PROPERTIES. 11
cumstances a purple solution is obtained. On this account the adjective
“ purpling ” has been applied to the chromogen in the title of this
paper. The purple compound which is formed in aqueous solution
is identical with the red compound which is formed in acetic-ether
solution. It may be completely removed from aqueous solutions by
shaking with acetic ether. In water it is purple, in organic solvents
red. The red compound is too unstable to be obtained in a solid state
by the evaporation of its acetic-ether solution,
When the chromogen or one of its oxidation products is treated
with an alkali a salt is formed. In striking contrast to the series of
acids, the salts are all totally insoluble in acetic ether or chloroform,
but very soluble in water. The salts of both the chromogen and its red
oxidation product are deep green in aqueous solution. If a solution
of the chromogen or of the red compound in acetic ether is floated on
dilute ammonium hydroxid the corresponding ammonium salt is
formed at the juncture of the two liquids. After the layers are
shaken together the deep-green aqueous salt solution will settle out,
leaving the acetic ether colorless. By immediately acidifying the
mixture and shaking it the process may be reversed.
The ammonium salt of the chromogen in aqueous solution absorbs
oxygen from the air with avidity, passing first, without visible change,
to the similar salt of the red oxidation product. If the salt is acidi-
fied at this stage the red compound is obtained, and the chromogen
itself may be recovered therefrom by reduction with zinc. On longer
standing, however, the green salt solution becomes brown. If it is
then acidified and shaken with acetic ether a brown acetic-ether solu-
tion is obtained, which to all appearances contains the same brown
oxidation product of the chromogen that results when the chromogen
is directly oxidized beyond the red stage by nitric acid.
Dr. P. G. Nutting, of the Bureau of Standards, has kindly photo-
eraphed and absorption spectra of the chromogen, the wine-red oxida-
tion product, and the two green alkali salts. From an examination
of the negatives he has reported as follows:
The yellow chromogen solution, 1: 2,000 in acetic ether, 21 millimeters thick,
shows complete absorption of the ultra-violet. Transmission begins gradually
at 0.39, just at the limit of the visible violet, and increases steadily to full value
in the greenish blue at wave length 0.52.
The red oxidized chromogen, 1: 2,000 in acetic ether, 21 millimeters thick,
shows complete absorption of violet, blue, and green, gradually increasing
transmission through the yellow and orange, and full transmission of the red.
Transmission begins at wave length 0.55 in the greenish yellow and becomes
full at 0.63 in the red.
The green ammonium Salts, 1: 4,000 and 1: 40,000 in aqueous solution, showed
absorption so nearly nonselective that no determinations were made upon them.
When the chromogen is oxidized by plant oxidases it need not be
protected from the oxidant by solution in a solvent immiscible with
264
12 THE PURPLING CHROMOGEN OF A HAWAIIAN DIOSCOREA.
water. Potato juice (oxidase), prepared by grinding potato parings
and pressing through raw silk, actively oxidizes chromogen, which is
added in alcoholic solution, first, to the wine-red compound and then
to the more highly oxidized brown compound. The oxidation by
potato juice is accompanied by measurable oxygen absorption from
the air. The potato oxidase is much more active when oxygen is sup-
plied by hydrogen peroxid. Horse-radish juice alone (peroxidase)
fails to oxidize, even upon prolonged shaking, as is shown by the fact
that the chromogen may be recovered unchanged in color, and by
the additional fact that there is no measurable oxygen absorption.
The tests for oxygen absorption were conducted with the apparatus
designed and described by Bunzel.*. Hydrogen peroxid alone does
not oxidize the chromogen, but in the presence of hydrogen peroxid
horse-radish juice is a powerful oxidant, carrying the oxidation
almost instantaneously to the wine-red stage, at which it halts. The
chromogen is as sensitive in its reactions to oxidases and peroxidases
as guaiac resin,
Juice expressed from the Dioscorea tubers given rather unsatisfac-
tory results as an oxidase solution because of its highly mucilaginous
nature and because of the large quantity of easily oxidized tannin
which it contains. Nevertheless, juice pressed from parings by a
hydraulic press readily oxidized the chromogen in the presence of
hydrogen peroxid to the same wine-red compound which was obtained.
when other plant juices or inorganic oxidants were used. The pres-
ence of hydrogen peroxid seemed to be essential to the reaction; that
is, to use the terminology of Chodat, the tuber contains a peroxidase
but no associated oxygenase. A moderately active oxidase solution
was prepared from green tips of the stems of tubers which had
sprouted in the laboratory by mashing with water and filtering. This
solution gave (1) no reaction with chromogen alone, (2) an imme-
diate darkening and eventually a deep-purple color with chromogen
plus hydrogen peroxid, and (3) a more delayed but eventually pro-
nounced purpling with hydrogen peroxid alone. In the second case,
the purple-colored substance was of the usual wine-red color when
shaken out into acetic ether. In the third case, the apparent indica-
tion that a chromogen was present in the juice led to a further inves-
tigation of the stems in the hope of finding in what form, if any, the
chromogen of the tubers was translocated to the growing parts.
Several tubers were allowed to sprout in a dark chamber until the
white etiolated shoots were 6 inches to a foot in length. These were
cut into small pieces, discarding only the purple nodes and rudimen-
tary leaves, and dropped into boiling acetic ether. The material thus
1Bunzel, H. H. The Measurement of the Oxidase Content of Plant Juices, Bulletin
238, Bureau of Plant Industry. 1912.
264
EXISTENCE OF A SECOND CHROMOGEN. 13
killed was ground in a mortar with the acetic ether and the golution
filtered. The resulting pale-yellow solution was shaken with water
and the two layers separately examined. The acetic-ether layer con-
tained no chromogen which could be demonstrated by oxidation with
dilute fuming nitric acid or by the formation of a characteristic
alkali salt. The colorless water layer gave an immediate darkening
(green passing quickly to brown) with ammonia, and became rose
pink on the addition of acetic acid. The color obtained when the
solution was treated with dilute fuming nitric acid was of the same
intensity as that obtained with acetic acid, and it was therefore con-
cluded that the substance giving the reactions was the water-soluble
anthocyanin of the stems. The rose-pink compound formed by acidi-
fying the solution could not be shaken out into organic solvents.
Since the chromogen of the tuber is insoluble in water it would be
surprising to find it in the stem. If, as seems likely, the chromogen
is concerned with the formation of the water-soluble anthocyanin,
the change to a soluble compound takes place in the tuber. In this
connection it should be pointed out that no difference could be ob-
served between the water-soluble anthocyanin of the stems and that
of the tubers. (See p. 8.) The purpling of stem juice with hydro-
gen peroxid alone would seem to be due to the slight acidity of com-
mercial solutions of this reagent.
EXISTENCE OF A SECOND CHROMOGEN.
It remains to call attention to a second chromogen which occurs in
the tubers. The precipitate obtained when the ammonia-greening
chromogen was treated with lead acetate (see p. 9) was dried and
triturated with acetic ether slightly acidified with acetic acid. A
small part of the precipitate was thereby decomposed. After filtra-
tion the filtrate was neutralized by shaking. with bipotassium hydro-
gen phosphate and washed with water. Precipitation with lead
acetate and recovery by suspension in acidified acetic ether were re-
peated, and the chromogen precipitated in a relatively pure condition
by adding petroleum ether. It formed a brown resinous mass, similar
in appearance to the ammonia-greening chromogen already described.
Its yellow solution in acetic ether yielded a water-soluble purple
ammonium salt on shaking with dilute ammonium hydroxid. This
salt passed entirely into the ammoniacal aqueous layer, leaving the
acetic ether colorless. On standing, it quickly oxidized to a brown
compound, but if reacidified immediately after its formation the
vellow unchanged chromogen could be recovered in the acetic ether.
This ammonia-purpling chromogen gives an intensely red compound
when oxidized by shaking its acetic-ether solution either with greatly
diluted fuming nitric acid or with horse-radish peroxidase + hydro-
264
14. THE PURPLING CHROMOGEN OF A HAWAIIAN DIOSCORBA.
gen peroxid. The ammonium salt of the red oxidation product is a
purple compound, insoluble in both water and acetic ether.
NOMENCLATURE.
It is proposed to designate the ammonia-greening chromogen of
the Dioscorea tuber as rhodochlorogen. In so doing, the precedent
set by Reinke’ is followed. This author gave the name rhodogen
(formed on the analogy of the word chromogen) to the colorless sub-
stance from which on oxidation he obtained the red coloring matter of
beets. The word “ rhodochlorogen” (from 6ddov, rose, yAwpdc, green,
and yevjc, producing) is formed with reference to the colors of the
oxidation product and the alkali salts. It may be used as a generic
designation for chromogens which appear to be related to the
ammonia-greening anthocyanins, until the chemistry of these com-
pounds is better understood. The ammonia-purpling chromogen of
the Dioscorea tuber is hardly well enough established as a chemical
individual to warrant applying even a temporary designation to it.
POSSIBLE RELATIONSHIP WITH THE ANTHOCYANINS.
In order to fix the characteristics of the two Dioscorea chromogens
in mind, they are contrasted briefly in Table I before proceeding with
a discussion of their possible relationship to the anthocyanins.
TABLE I.—Comparison of two Dioscorea chromogens.
Rhodochlorogen. Ammonia-purpling chromogen.
Solution in acetic ether.:..2.:.:.....--.: Y ellow ack }clvre dun Spade ack 3 - Yellow.
Ammonium salt formed in acetic ether. .| Insoluble green precipitate..... Purple, partly soluble.
Ammonium salt shaken into water...... Deep-green solution..........- Purple solution.
Water solution of ammonium salt acidi- | Purple acetic-ether solution...) Insoluble purple precipitate.
fied after partial spontaneous oxida-
tion and shaken with acetic ether.
Oxidized by peroxidase + hydrogen per- | Purple solution. .......-.....- _ Red-purple solution.
oxid in very dilute alcoholic solution.
Oxidized by nitric acid in acetic-ether | Wine-red solution. ..........-.. Brownish red solution, even-
solution. tually a purple precipitate.
Ammonium salt of oxidation product | Insoluble green precipitate..... Insoluble purple precipitate.
formed in acetic ether.
Ammonium salt of oxidation product in | Deep-green solution........... Insoluble purple precipitate.
water.
The similarity of these chromogens to the anthocyanins at once
suggests itself. There is present in the stems and petioles of the
bitter yam a water-soluble cell-sap color which belongs to the
1 Reinke, J. Ein Beitrag zur Kenntnis leicht oxydirbaren Verbindungen des Pflanzen-
kirpers. Zeitschrift fiir Physiologische Chemie, vol. 6, 1882, p. 263. “* * * in
der lebenden Pflanzenzelle leicht oxydirbare Substanzen vorhanden sind, welche begierig
atmosphiirischen Sauerstoff apzieben, uvd mit demselben Oxydationsprodukte bilden.” In
referring to the chromogen which produces the red coloring matter of beets, Reinke says,
‘“ Weil der Sauerstoff der atmosphiirischen Luft diese Substanz zu einem roten Farbstoff
zu oxydiren vermag, will ich dieselbe als Rhodogen bezeichnen * * = *,”
264
,
.
'
POSSIBLE RELATIONSHIP WITH THE ANTHOCYANINS. 15
apex of the petioles (see fig. 1). It forms green salts with alkalis,
ammonia-greening class of anthocyanins. It is of a very deep purple-
red color and is particularly conspicuous in the enlarged base and
which on standing spontaneously oxidize to brown compounds. It
may be reduced by boiling with zinc dust to a yellow compound,
from which the anthocyanin is regenerated by oxidation. (The
oxidation of the yellow reduction product takes place spontaneously
when its solution is exposed to the air.) This yellow reduction com-
pound of the anthocyanin is
strictly comparable to rhodo-
chlorogen in that it forms
green alkali salts. Rhodo-
chlorogen oxidizes to a purple
or red compound, depending
upon the solvent. The yellow
reduction product reoxidizes
to purple or reddish purple
anthocyanin. Thus far, the
similarity of the two com-
pounds is perfect. Just as
the yellow rhodochlorogen
and its purple or red oxida-
tion product both form green
salts, so the purple anthocya-
nin and its yellow reduction
compound both form green
salts. The two pairs of com-
pounds are strikingly unlike
only in their solubility rela-
tions, rhodochlorogen and its
red oxidation compound be-
ing soluble in acetic ether or
ehlorotorm but not in water, Fic. 1—Leaf of Dioscorea: showing petiole en-
whereas the anthocyanin and laxBerl at base and apex, immature aerial]
; : tuber, and bases of racemes. (Reduced to
its yellow reductioncompound __ one-half diameter.)
are soluble in water but not in
acetic ether or chloroform. The anthocyanin has not been isolated
and is therefore not known to be a glucosid, but it seems probable,
in view of Grafe’s investigations, that the two water-soluble com-
pounds are glucosids of the two which are not water soluble. Even
if they are not related in this way, there is strong circumstantial evi-
dence that the chromophoric nucleus in the molecules of rhodochloro-
gen and anthocyanin is identical and therefore that the two sub-
stances are in some way genetically related in the plant metabolism.
264
-_
16 THE PURPLING CHROMOGEN OF A HAWAIIAN DIOSCOREA.
In this connection attention should be called to the fact that Grafe?
has resolved the ammonia-greening anthocyanin of Althaea rosea
into two components. One, a water-soluble sap color of the usual
type, is a glucosid of the formula C,,H,,O,,.. The other, insoluble
in water but soluble in absolute alcohol, is not a gucosid. Its for-
mula C,,H,,O, differs from that of the glucosid by a molecule of
water and a molecule of glucose. In their color reactions the two
components are similar, and there is little doubt that they contain the
same chromophoric nucleus. The nonglucosid, which is insoluble in
water, greatly resembles the red oxidation product of rhodochloregen
in that it may be reduced to a yellow compound which seems to be
analogous to rhodochlorogen itself.’
Glan*® had already shown before Grafe took up his work on the
mallow pigments that the water-soluble anthocyanin of
‘
aa bia yTier ew Oia
P ‘ 7
’ ,
. 7 A ci 2st
s* ° ¢
wis re aoe Las
x ' if et) ‘
‘ ae _~ o< ~ sy
ei ise
aS rial
ad _
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. la J . «
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Issued December 16, 1912,
U. S. DEPARTMENT OF AGRICULTURE,
BUREAU OF PLANT INDUSTRY—BULLETIN NO. 265.
B, T. GALLOWAY, Chief of Bureau. ;
SOME FACTORS INFLUENCING THE
EFFICIENCY OF BORDEAUX
: MIXTURE.
BY
LON A. HAWKINS,
Scientific Assistant, Fruit-Disease Investigations.
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CONTENTS.
4 Page.
I tests ode ca oa hs oowawm se dean oo ated eat ne ne sees ake ee 7
nent OF DOrdeaux Mixture... 2-6. Gs. 253. 5- oe Go de ee eee anaes 7
@aeparaton of Bordeaux mixture...5.....2.225. 5.25 s05 ee eee ee eee eee nes 8
Suemmcrintente 1 preparation... 2... seeped esses edace we = ee eae 10
Effect of different methods of mixing on the rate of subsidence of the
BORURONBONE. 52 .osiia Sree! ay. ee a aE. DIRS Qe 10
Effect of varying amounts of agitation on the subsidence of the sus-
a Gen 25 oo le saws oo ee Se eee Sot ee Se ae 11
es Experiments with concentrated lime poured into dilute copper
: ‘ Mitpaber reine p iss Lewes BeOS ARO ee 11
a Experiments with concentrated copper sulphate poured into
ee atte Winer se ah 26). pean OL EU ae Ae 13
ce Discussion of effects of agitation.......-......-..-..2.-2.222222-- 13
, Adherence of Bordeaux mixture with and without added adhesives.......... 16
a Historical review of work on adherence............----------+---eeeee eee 16
‘ Peperiments on. adhesiveness. . 2.2.2.2 59802. oye ee 17
7 / Experiments on adhesiveness to grape leaves..........------.------- 18
e Experiments on adhesiveness to grape berries..........-..--.------- 20
a. Surface tensions of mixtures used........-...-.....--..2-0------ 21
A laboratory method for comparing the efficiency of added adhesives. . 23
Discussion of the results of the experiments.....................------------ 26
EM SEESEONGERE SUID cc Es OSs, a a i ere Se 28
265
ILLUSTRATIONS.
Page. .
Fia. 1. Diagram showing the effect of varying the amount of agitation of
Bordeaux mixture when the concentrated lime is added to the
diluted copper-sulphate solution. 5... ene ence ecielee so 12
2. Diagram showing the effect of varying the amount of agitation of
Bordeaux mixture when the concentrated copper-sulphate solution is
added to the: diluted lime... i. .Dodic2.00.. 0. 5¥. eb ope ee 14
3. Sketch of apparatus used for measuring the depression of the suriace
films of Bordeaux mixture by the bloom of the grape...........---- 24
4, Diagram which compares the average depressions of the surface films
seen in the horizontal microscope of Bordeaux mixture without
added adhesive and Bordeaux mixture with 2 pounds of rosin-fishoil
soap to 50 gallons of mixture... ....5--s-s00 56.20.00. see eee 25
265
6
B. P. I.—778.
SOME FACTORS INFLUENCING THE EFFI-
CIENCY OF BORDEAUX MIXTURE.
—
INTRODUCTION.
_ The efficiency of Bordeaux mixture in preventing certain diseases”
which attack the young aerial portions of plants is dependent on
several factors. Not the least among these is uniformity in the dis-
tribution of the copper compound throughout the liquid medium
when the mixture is applied. That this is of importance is very
apparent, for if the copper compound has settled out, even to a limited
degree, part of the plant will receive a heavy coating of the fungi-
cide, while other portions may receive none and thus be liable to
4 infection by the fungus. Another important factor is adhesiveness,
‘: as it is obviously necessary for the fungicide to adhere to the sus-
ceptible portions of the plant if they are to be protected from fungous
disease. With these requirements for efficiency in mind the questions
naturally arise, By what methods can the most uniform distribution
of the copper compound in the medium be obtained, and how can the
adhesiveness of the mixture to the susceptible parts of the plants be
increased? The present investigation deals with these two questions.
COMPOSITION OF BORDEAUX MIXTURE.
. Bordeaux mixture is made up of copper sulphate and calcium
__ hydroxid, and the rate of subsidence of the colloidal suspension of
the precipitate which results from the interaction of these substances
is partly dependent on the manner in which the two components are
brought together. It is not necessary here to go into a detailed dis-
cussion of the chemical reactions that take place when copper sulphate
and calcium hydroxid are brought together. They have been studied
by Swingle,t Chester,? Sostegni,? Pickering,* and others, with various
1$wingle, Walter T. .Bordeaux Mixture: Its Chemistry, Physical Properties, and Toxic
Effects on Fungi and Alge. Bulletin 9, Division of Vegetable Physiology and Pathology,
U. S. Dept. of Agriculture. 1896.
2Chester, F. D. Copper Salts as Fungicides. Journal of Mycology, vol. 6, 1890, pp.
21-24.
*Sostegni, Livio. Sulla Composizione Chimica della Cosi detta Poltiglia Bordolese. Le
Stazioni Sperimentali Agrarie Italiane, vol. 19, 1890, pp. 129-141.
“ “Pickering, Spencer U. Eleventh Report of the Woburn Experimental Fruit Farm.
‘a 1910. ‘
a 265 rs
8 FACTORS INFLUENCING EFFICIENCY OF BORDEAUX MIXTURE. _
conclusions as to the nature of the compounds formed. It is gen-
erally agreed that the insoluble copper compound of Bordeaux mix-
ture, whether copper hydroxid, basic sulphate of copper, or boththese
compounds, is in colloidal suspension in a saturated or nearly satu-
rated solution of calcium sulphate and calcium hydroxid. 7
PREPARATION OF BORDEAUX MIXTURE.
Different authors have recommended various methods for the prep-
aration of Bordeaux mixture, with a view of obtaining the most eco-
nomical and effective mixture. Millardet,1 in describing the making
of Bordeaux mixture for the first time, says:
Dans 100 litres d’eau quelconque (de puits, de pluie, ou de riviére) on fait
dissoudre 8 kilos de sulfate de cuivre du commerce, D’un autre cété, on fait,
avec 30 litres d’eau et 15 kilos de chaux grasse, en pierres, un lait de chaux
qu’on mélange 4 la solution de sulfate de cuivre.
This method of mixing, with the same formula, was adopted in
America, having been first published by Scribner? in 1886. Two
years later Scribner* recommended 4 pounds of copper sulphate and
a, like quantity of lime in 22 gallons of mixture, while Galloway®
the same year recommended a formula of.6 pounds of copper sul-
phate and 4 pounds of stone lime to 22 gallons of water. Waite® in
1893 obtained good results in spraying for pear leaf-blight by using
6 pounds of copper sulphate to 50 gallons of water with just sufficient
lime to react with the copper sulphate. In the same article this
writer recommends the use of a stock solution of copper sulphate and
a stock mixture of lime in the preparation of the fungicide. With
these formulas, the method of preparation was to pour the calcium
hydroxid into the copper-sulphate solution. In 1896 Galloway’ rec-
ommended the use of two tubs, in which the copper sulphate and
lime were separately diluted, each to half the volume of the Bordeaux
mixture required. From these tubs the two solutions were poured
simultaneously into a barrel: In the same article he recommended
>
eth
1Millardet, A. Journal d’Agriculture et d’Horticulture de la Gironda, May 1, 1885.
Nory.—This publication was not at hand and the quotation given was taken from the
same writer’s paper, entitled ‘“‘ Sur l’histoire du traitement du mildiou par le sulfate de
Cuivre,” Journal d’Agriculture Pratique, vol. 49, pt. 2, 1885, pp. 801-805, in which Mil-
lardet quotes directly from his former paper in describing the method of preparing
Bordeaux mixture. :
2Scribner, F. Lamson. Report on the Mycological Section, in the Report of the Com-
missioner of Agriculture for 1886, p. 100.
8 Report on the Fungous Diseases of the Grapevine. Bulletin 2, Section of
Plant Pathology, Botanical Division, U. 8S. Dept. of Agriculture, 1886, p. 16.
‘ Fungicides or Remedies for Plant Diseases. Circular 5, Section of Vegetable
Pathology, Botanical Division, U. 8. Dept. of Agriculture, 1888.
5Galloway, B. T. Treatment of Black Rot of the Grape. Circular 6, Section of
Vegetable Pathology, Botanical Division, U. S. Dept. of Agriculture, 1888, p. 2.
®Waite, M. B. Treatment of Pear Leaf-Blight in the Orchard. Journal of Mycology,
7, 1894, pp. 333-338.
7Galloway, B. T. Spraying for Fruit Diseases, Farmers’ Bulletin 38, U. 8. Dept. of
Agriculture, 1896, p. 6.
265
PREPARATION OF BORDEAUX MIXTURE. 9
6 pounds of copper sulphate and 4 pounds of lime to 50 gallons of
mixture. This last method of mixing Bordeaux has been recom-
mended by the investigators in the Department of Agriculture and
most of the agricultural experiment-station workers in the United
States since that time. Some of the experiment stations, however,
recommend the pouring of one component into the other, as shown
by the publication of Woods and Hanson,’ Green, Selby, and Gos-
sard,? and Smith and Smith.* Kelhofer* in an account of his in-
vestigations on the preparation of Bordeaux mixture says:
Die gréssten Niederschlige erzielen wir demnach bei langsamem (portionen-
weisem) Zusatz der Kupfervitrioll6sung zur Kalkmilch. Annihernd ebenso
giinstige Resultate werden erhalten, wenn man die Kalkmilch rasch zur Kupfer-
vitriolldsung giesst.
The copper sulphate and lime of Kelhofer’s preparations were both
diluted to the same volume. Two series of experiments were carried
out, in one of which this volume was one-half that of the fungicide
required and in the other one-fourth. Kelhofer®-* also added with
good results small quantities of cane sugar to retard the rate of sub-
sidence of the suspension. Kulisch’ repeated some of Kelhofer’s
experiments with like results. Pickering,® in making common Bor-
deaux mixture, recommends the use of calcium hydroxid as dilute
as possible to make the required quantity and the copper sulphate in
concentrated solution. The copper sulphate is poured into the cal-
cium hydroxid with very little stirring. An examination of the liter-
ature of this subject shows that the methods recommended for the
preparation of a colloidal suspension of the copper compound which
‘settles out slowly are rather varied. The problem of making a sus-
pension which subsides slowly then resolves itself into testing the
methods of mixing recommended by the different investigators to
determine their comparative efficiency. Accordingly, to determine
the effect on the rate of subsidence of the suspensions of some of these
1 Woods, Charles D., and Hansen, H. H. Paris Green Bordeaux Mixture. Bulletin
154, Maine Agricultural Experiment Station, April, 1908.
2Green, W. J., Selby, A. D., and Gossard, H. A. Spray Calendar. Bulletin 232, Ohio
Agricultural Experiment Station, 1911.
8 Smith, R E., and Smith, Elizabeth H. Bulletin 218, Agricultural Experiment Station
of the University of California, 1911, p. 1185.
4Kelhofer, W. Versuch iiber die Herstellung der Bordeauxbriihe. Jahresbericht der
Deutsch-Schweizerischen Versuchstation und Schule fiir Obst-Wein- und Gartenbau, vol.
8, 1897-98, p. 65.
5 Kelhofer, W. Versuche iiber die Beeinflussung der Haltbarkeit der Bordeaubriihe
durch Zusitze. Jahresbericht, der Deutsch-Schweizerischen Versuchstation und Schule fiir
Obst-Wein- und Gartenbau, vol. 9, 1898-99, pp. 87-92.
6 Ueber einige Gesichtspunkte bei der Herstellung der Bordeauxbriihe. Zeit-
schrift fiir Pflanzenkrankheiten, vol. 18. Internationaler Phytopathologischer Dienst, vol.
1, no. 3, 1908, pp. 65-78.
7Kulisch, P. Die Darstellung haltbarer Kupferbriihen zur Bekimpfung der Peronospora.
Zeitschrift fiir Pflanzenkrankheiten, vol. 21, 1911, pp. 382-384.
§ Pickering, Spencer U. Op. cit., p. 56.
61566°—Bul. 265—12——2
Se A Boat 3
eh Bey:
10 FACTORS INFLUENCING EFFICIENCY OF BORDEAUX MIXTURE. —
methods of preparing Bordeaux mixture the investigations described
in the first part of this paper were planned and carried out.
EXPERIMENTS IN PREPARATION.
For the greater part of the investigation the copper sulphate and
lime used were what is commonly known as chemically pure. Dis-
tilled water was used in these preparations. Later, a number of the
series were repeated in order to approach commercial conditions as
closely as possible, using a good grade of common stone lime, com-
mercial copper sulphate, and tap water. The mixtures were prepared
in glass-stoppered cylinders of 1-liter capacity graduated to divisions
of 10 cubic centimeters. To prepare Bordeaux mixture by allowing
the two diluted components to flow simultaneously into the container,
two burettes of 1,000 cubic centimeters capacity were placed side by
side, with the outlets connected by rubber tubes provided with
pinch cocks to a single Y tube, the lower arm of which was so ar-
ranged as to project into the neck of the glass cylinder. The proper
quantity of calcium hydroxid, made by slacking 3.75 grams of cal-
cium oxid, was placed in one burette and diluted to half a liter,
while in the other was placed a solution of the same volume, con-
taining 5 grams of copper sulphate.
DISCUSSION OF THE RESULTS OF THE EXPERIMENTS. 27
_ Of the adhesive compounds added to Bordeaux mixture, the rosin-
- fishoil soap proved to be most effective on the grape berries—much
- more effective, in fact, than fishoil soap without the rosin. From this
- fact it seems probable that the adhesiveness is largely due to the
rosin present. It is stated by various writers that the addition of a
- small quantity of soap to Bordeaux mixture could be of no particu-
lar benefit, as the soap would be precipitated as an insoluble calcium
soap by the excess of calcium present in the mixture. Good results
could therefore be expected only when a considerable quantity of
soap was added. This, of course, may be true of certain kinds of
soap, but in this investigation considerable benefit was derived from
the addition of relatively small quantities of soap. Even the fishoil
soap materially increased the adhesiveness over Bordeaux mixture
without added adhesives.
In the treatment of the black-rot. of the grape good results have
been obtained in many cases by using Bordeaux mixture without
added adhesives. When we consider this fact in connection with the
evidence brought forth in the present investigation, that Bordeaux
mixture without added adhesives does not adhere to the grape berry
- im any appreciable quantity, it seems probable that the protection is
_ due to reducing the sources from which infection comes to the berry.
__ By protecting the foliage from infection the possibility of secondary
_ infection from the foliage to the fruit may be eliminated to a consid-
_ erable extent. Covering the stems of bunches of grapes with the
_ fungicide seems to be another means by which infection may be kept
_ from the grapes. The writer has observed numerous instances of
black-rot infection on bunches of grapes which had been bagged six
weeks or more. In these instances spores probably washed down the
stems in drops of water, as the only openings in the bags were imme-
_ diately around the stems.
Though good results were obtained by the addition of glue to
_ Bordeaux mixture, its cost (about 12 cents a pound) prohibits its
__use in commercial work in place of rosin-fishoil soap. When glue is
' added to alkaline Bordeaux mixture, part of the copper combines
_ with the glue, forming a soluble compound bright purple in color.
_ Itis probable that much of the copper found on the grapes from this
plat was in this form. As it is soluble in water, this protective
covering might not remain on the berries as long as the insoluble
_ precipitates in the mixtures with the soap.
It is difficult to see in just what way ferrous sulphate could be
expected to influence the adhesiveness of Bordeaux mixture. On
_ the addition of this compound to a solution of calcium hydroxid,
| ferrous hydroxid immediately precipitates out, and none of the
. 265
a Bc
4 a |
28 FACTORS INFLUENCING EFFICIENCY OF BORDEAUX MIXTURE. =
ferrous sulphate remains in solution. It was found to be worthless
as an adhesive for use on grapes.
In the plats sprayed with 3-2-50 Bordeaux mixture, to which was
added 2 pounds of rosin-fishoil soap, a considerable quantity of cop-
per was found on the grapes. This formula has given excellent
practical result and should prove much more effective than the
mixtures containing more copper but without the adhesive.
a! CER ge tony fo Ceione* Ua, 4
CONCLUSION.
It has been shown in these investigations that a Bordeaux mixture __
in which the suspension of the copper compound setéles out slowly ‘
may be prepared by adding the concentrated calcium hydroxid to
the diluted copper-sulphate solution or vice versa, provided the mix- :
ture is sufficiently agitated. Practically as good results were ob-
tained with these methods of preparation as by diluting the two com-
ponents in separate vessels and pouring them simultaneously into a
third, as recommended by Galloway in 1896.
It is to be remembered that in preparing Bordeaux mixture, by
pouring one of the components in concentrated form into the other __
diluted to nearly the required volume, the resulting mixture must be
thoroughly agitated. The agitation necessary for preparing Bor-
deaux mixture with a low rate of subsidence by this method could
hardly be obtained in practice except by means of a power outfit
provided with a good agitator. This method of mixing is not de-
signed to replace the old gravity method with its elevated platform,
but offers a convenient substitute where for any reason the gravity
method is impracticable.
In the experiments on the adhesiveness of certain Bordeaux mix-
tures and the relative value of certain adhesive compounds it was
shown that by determining the quantity of copper retained on the
leaves sprayed with the different mixtures the addition of rosin-
fishoil soap slightly increases the adhesiveness of the mixture. In
similar experiments on grape berries it was shown that the adhesive-
ness of the fungicide could be materially increased by the addition
of certain adhesive compounds. Two pounds of rosin-fishoil soap
to 50 gallons of spray mixture was the most valuable of any added
adhesive, ground glue was second, 1 pound of rosin-fishoil soap to
50 gallons of mixture was third, and fishoil soap was fourth. Fer-
rous sulphate did not increase the adhesiveness of the Bordeaux mix-
ture, as no appreciable quantity of copper adhered to the grape
berries where the Bordeaux mixture to which the ferrous sulphate
had been added was used. No appreciable quantity of copper was
found on the grape berries from the plats sprayed with Bordeaux
mixture without added adhesives. From the experiments on grape
265
i ein CONCLUSION. 29
berries with adhesives it may be concluded that the use of an adhesive
“compound i is necessary to make the fungicide adhere to the bloom-
overed grapes. Two pounds of rosin-fishoil soap to 50 gallons of
Retixture gives the best results and is recommended as the most eco-
; - nomical and efficient adhesive for use on grape berries. From the re-
4 sults obtained with 3-2-50 Bordeaux mixture, with the addition of
4 soap, it seems probable that a mixture containing this quantity of
_ copper sulphate would be effective when a good adhesive is used. A
- laboratory method of approximating the relative adhesiveness of
_ these fungicides to grapes was developed.
265
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Issued February 21, 1913.
U. S. DEPARTMENT OF AGRICULTURE.
BUREAU OF PLANT INDUSTRY—BULLETIN NO. 266.
B. T. GALLOWAY, Chief of Bureau, \
THE DESTRUCTION OF CELLULOSE BY BACTERIA
AND FILAMENTOUS FUNGI.
BY
I. G. McBETH, Physiologist,
AND
F. M. SCALES, Assistant Soil Mycologist,
Sowl-Bacteriology and Plant-Nutrition
Investigations.
WASHINGTON>
GOVERNMENT PRINTING OFFICE,
1913,
BUREAU OF PLANT INDUSTRY.
Chief of Bureau, BEVERLY T. GALLOWAY.
Assistant Chief of Bureau, WILLIAM A. TAYLOR.
Editor, J. E. ROCKWELL.
Chief Clerk, JAMES E. JONES.
SOIL-BACTERIOLOGY AND PLANT-NUTRITION INVESTIGATIONS,
SCIENTIFIC STAFF.
Karl F. Kellerman, Physiologist in Charge.
y;
1. G. McBeth, Physiologist.
¥. M. Scales, Assistant Soil Mycologist.
.C. Wright, F. L. Goll, Edna H. Fawcett, and L. T. Leonard, Scientific Assistants.
R. Smith, Willis, L. Hurd, and A. P. Harrison, Laboratory Aids.
266
ADDITIONAL COPIES of this publication
may be procured from the SUPERINTEND-
ENT OF ete ren, Government Printing
Office, Washington, D. C., at 10 cents per copy
LETTER OF TRANSMITTAL.
U.S. DEPARTMENT OF AGRICULTURE,
BuREAU OF PLANT INDUSTRY,
OFFICE OF THE CHIEF,
Washington, D. C., September 12, 1912.
Sir: I have the honor to transmit herewith and to recommend for
publication as Bulletin No. 266 of the series of this Bureau a manu-
script entitled ‘‘The Destruction of Cellulose by Bacteria and Fila-
mentous Fungi.” This paper was prepared by Messrs. I. G. McBeth,
Physiologist, and F. M. Scales, Assistant Soil Mycologist, of the
Office of Soil-Bacteriology and Plant-Nutrition Investigations, and
has been submitted by the Physiologist in Charge with a view to
publication. |
New species of cellulose-destroying organisms are described, as
well as special methods and new culture media adapted to their
isolation and identification.
Respectfully, B. T. GALLowaAyY,
Chief of Bureau.
Hon. JAMEs WILSON,
Secretary of Agriculture.
266 8
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CONTENT 3.
Page.
IEIORC 0 oo Ss SRC cat. So a ie el ate ae klga toes eae 7
ES Sino nso do aie < woe Pp Ae Reo on aa a ah 9
Historical review of investigations of the destruction of cellulose...........-- 10
Bacteriological and chemical investigations.............----------------- 10
Peveswations with filamentous fungi*...< 1.2... .. 2-525. 5. i. 2 eke eek 23
_pocs and Culture media employed... -.2.--¢..--.--se272. ----225 2S kee 25
eer PAT cc ke Od Se OE a 0 Ae co 27
MUNN oie Wien Sele a ga eS... 2 28
RRRDEN ess oka es 2S es dare w x wn LS eS oon a a we 28
pe Green SSeS. At a AS. 2 Doe ee a» Ls SU 28
The occurrence and general characteristics of cellulose-destroying bacteria in
RM. Shane EIS 2) LL ee ee 29
Descriptions of cellulose-destroying bacteria........:....-..--..-2.2.2.----- 30
TET a ee aie See a Sm ge PEPE > ee, ELE pg Pa 30
RE gee 0 Ieee BA Gh tea hc cy J = ee ae fe 32
PNG AMR POS Party. Stas hae ae ao. SR ae 35
Peemmarnnonas pupcrebas 2." -. 2250.08. 20 = sapere se 37
EEE MERON a5 Sr cscs fo a icine Pls SI SERIES = ae 3 SR 39
Cellulose destruction by filamentous fungi....................-----------.0-- 41
Products resulting from the destruction of cellulose by bacteria..........-...-- 43
) NE er csr 5. tent cA yee, aA RR se wk a 45
D a Rael URS at ARR age cn sy Sts 9. ee 47
SIMMER NTT OR PAGLOS. ons wcrc’. 5 ~~~ a ood e oe eden ea s+ --- see ee ene edee 52
266 5
[LL ERA aS,
Page.
PuaTE I. Colonies and vegetative cells of Bacterium liquatum and Pseudo-
monhassuberetus ...2 0d les.k. ci pese newts 1ewcurceis eee 52
II. Colonies and vegetative cells of Bacterium fimiand Bacillusbibulus.. 52
III. Colonies, spores, vegetative cells, and involution forms of Bacillus
CYTASETIB nn. on nce cins nisms aos aanne = inane © «sis een 52
IV. Vegetative cells of Bacillus bibulus and Bacillus cytaseus, showing
BG ise aS cn «www annem jninicje ais ee ie a Seam gee 52
266
PROGATOMY WOE.
Of the phenomena relating to the art of argiculture none is more
interesting or more vitally important than the formation of starch,
sugar, cellulose, and similar compounds by green plants. The
action of chlorophyll, as yet unexplained, by which plants utilize the
energy of the sunlight to synthesize carbon dioxid and water into
carbohydrates is logically the most fundamental question of plant
physiology, for it is the enormous quantity of potential energy thus
accumulated that directly or indirectly makes possible the continu-
ance of all vital processes. Not only animal life which, generally
speaking, is dependent upon plants for food supply, but even the
successful growth of crop plants is largely controlled by the decom-
posing carbonaceous material in the soil.
Both in the Eastern and Western States the natural maintenance
of the supply of available nitrogen is seldom considered when deter-
mining the most desirable system of farm management in any
region, yet, as scientific research and experience in the field agree in
showing, the soil itself may fix and render available to the crops
considerable quantities of nitrogen, which is the highest priced of
the plant foods if it be purchased as commercial fertilizer. Further-
more, as far as our experience extends, all of the fixation of atmos-
pheric nitrogen in the soil is dependent upon the growth of micro-
organisms which must have large quantities of soluble carbon com-
pounds for food. As by far the larger part of the carbonaceous
material added to the soil as dried roots, stubble, green manure,
etc., 1s cellulose, a substance which is unusually refractory and can
not be used as such for food by nitrogen-fixing organisms, the bio-
logical phenomena which transform the cellulose to soluble com-
pounds are obviously important. It is not only as a possible food
supply for nitrogen-fixing organisms, however, that a soil requires a
constantly replenished supply of cellulose. Its decomposition
under proper soil conditions and in association with the decay of
nitrogenous compounds makes possible the formation of the so-called
soil humus. The beneficial effects of the presence of indefinite
humic compounds upon the physical character and fertility of a
soil are generally recognized throughout the agricultural regions of
the United States. |
266 7
8 PREFATORY NOTE.
The gradual processes of decay that are depended upon to main-
tain many of the factors of a soil’s fertility are probably as com-
plex as the microflora and fauna of the living soil itself, and it is
chiefly through the discovery and comprehension of the essential
biological phenomena relating to the growth of plants that per-
manent improvement in crop production can be made possible.
The decomposition of cellulose is apparently one of the fundamental
questions of the decay of organic material, and though a subject of
research in foreign countries for many years, it has been but imper-
fectly understood. The investigations, therefore, of which this
bulletin is a progress report, are regarded as of unusual importance.
Karu F. KELLERMAN,
Physiologist in Charge.
OFFICE OF SorL-BACTERIOLOGY INVESTIGATIONS,
Washington, D. C., September 10, 1912.
266
B. P. I1.—779.
THE DESTRUCTION OF CELLULOSE BY BACTERIA
AND FILAMENTOUS FUNGI
INTRODUCTION.
The important functions of fission fungi is to dissolve and again
place in circulation the complex organic substances which have ceased
tolive. Without their activity the cycle of change to which all organic
matter is subject would come to a standstill and the food supply of
plants would soon be depleted. It is well known that through the
agency of micro-organisms all vegetable matter is gradually trans-
formed into the complex mixtures ordinarily known as humus and
that we are at least partially dependent upon the quantity and
quality of the humus compounds for the fertility of the soil.
It is true that numerous chemical researches have added materially
to our knowledge of these organic or humic compounds, but since the
biological processes involved are the dominant factors in determining
the manner in which complex organic substances are split up, a system-
atic study of the organisms which bring about the decomposition
of vegetable matter and the formation of plant food is imperative.
Vegetable substances may be roughly divided into two great classes,
nitrogenous and nonnitrogenous. In the decomposition of nitrog-
enous matter we are concerned chiefly with the fate of the nitrogen,
a part of which seems to be invariably returned to the atmosphere.
It is well known that this loss may be considerable. Fortunately,
however, nature has provided a means of restoring this lost nitrogen
through the activity of certain so-called nitrogen-fixing micro-
organisms. A study of these organisms and the conditions under
which they are able to fix nitrogen has shown that the process is con-
trolled in large measure by the available supply of organic carbon.
On examining plant tissues we find a large percentage of the carbon
content locked up in the celluloses; these are inert compounds which
resist the attacks of the ordinary putrefactive bacteria and until
broken down into simpler compounds are inaccessible to nitrogen-
fixing bacteria. Little is known of the biological processes involved
in the destruction of cellulose. It is true that many foreign investi-
gators have studied cellulose ferments, but generally the work has
62420°—Bull. 266—13——2 9
10 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
been done in a tentative way, and no suitable methods for isolating
these essential organisms have been worked out. Consequently, our
ideas of the number and nature of the cellulose ferments have been
very inadequate. Investigators have also devoted much attention
to the products resulting from the fermentation of cellulose, but they
apparently have been working with impure cultures and their con-
clusions are of doubtful value.
Believing that little progress could be made in the study of cellulose
decomposition until satisfactory methods for isolating these organisms
had been perfected, thus giving opportunity to learn something of
their cultural characteristics, we have endeavored to work out
methods to isolate and study them. The purpose of this bulletin is
to review briefly the work of earlier investigators, point out the
inadequancy of our present knowledge of cellulose fermentation, and
set forth the results obtained from our own studies in the hope that
they may be of value to other investigators.
HISTORICAL REVIEW OF INVESTIGATIONS OF THE DESTRUC-
TION OF CELLULOSE.!
BACTERIOLOGICAL AND CHEMICAL INVESTIGATIONS.
The fermentation of cellulose was first attributed to the activity
of microorganisms by E. Mitscherlich in 1850. He noted that when
slices of potato were immersed in water and held in a warm place
the cellulose, which constitutes the main portion of the cell walls,
was destroyed. First, the cells became separated from each other,
and soon afterward the walls were broken down and the starchy
material fell out. By filtering the solution and dropping in fresh
potato the fermenting process was greatly accelerated. Microscopic
examinations showed no trace of a mold growth, but Mitscherlich
observed swarms of vibrios, which he believed to be the active agents
of cellulose fermentation.
Four years later Haubner showed that it was impossible to recover
from the feces more than 50 per cent of the crude fiber fed to rumi-
nants. He obtained similar results with wood shavings which had
been treated with acid and alkali, and also with thoroughly washed
paper when fed with hay and bran to sheep. Haubner’s work was
soon confirmed by Henneberg and Stohmann. Through the experi-
ments of Hofmeister, Zuntz, Knierem, Weiske, Lehmann, and others
similar results were obtained with horses, sheep, goats, rabbits, etc.
Although undertaken primarily to determine the nutritive value of
crude fiber in foodstuffs, these investigations no doubt did much to
stimulate later investigation which sought to determine the causative
agent of cellulose fermentation.
268 1 For bibliography, see pages 47 to 50.
HISTORICAL REVIEW OF INVESTIGATIONS, 11
In 1865 Trecul undertook a study of microorganisms in macerated
plant tissues. He observed and described three forms, which he
; placed in a distinct genus, Amylobacter, and divided into three sub-
genera. This generic name was selected because these organisms
stained blue with iodin. He believed that starch or cellulose favored
the production of these bodies.
| For our early knowledge of cellulose fermentation we are much
indebted to the work of Popoff, who in 1875 first pointed out the
connection between cellulose fermentation and the formation of
methane. Methane had Jong been known to exist in sewers and
marshes and had been found in fermenting horse manure by Reiset
as early as 1856; however, no successful attempt had been made to
determine the source of the gas. For experimental material Popoff
used slime from the sewers of Strassburg. The material was mixed
with sufficient water to make a thick solution, poured into large
flasks, and preparations made to collect the gas over quicksilver.
An analysis of the gas showed that considerable quantities of methane
mixed with other gases had been produced. The optimum tempera-
ture for the gas formation was found to be 38° C. to 40° C.; at 45° C.
the activity was much weakened, and at 50° C. it came to a stand-
4 still; lower temperatures were also shown to be very unfavorable.
Popoff further showed that the fermentation process could be altered
at will by the addition of antiseptics. The next step was to show
that the formation of methane could result from the destruction of
pure cellulose. With this end in view a quantity of pure Swedish
filter paper was immersed in water and inoculated with a small
- quantity of sme known to contain the methane ferment. The paper
was destroyed and a large quantity of gas was formed, which on
examination proved to be a mixture of carbon dioxid, methane,
hydrogen, and nitrogen. The gases collected during the first two
weeks and again several weeks later were analyzed with the result
shown in Table I.
TaBLE I1.—Analyses of mixed gases formed by the decomposition of Swedish. filter paper.
Collected at the
end of—
Gases found in the mixture.
Two Several
weeks. weeks.
Per cent. | Per cent.
25.70
DTG e oo oe oe | ae ee ee ee ae MM 34. 07
Dat Talat So ect BES REN s BETS BA ge Be es Sop os eR REE BAe od, 14. 42 5 7 fp
SERRE SERRE a a Cie bien ee ee Oe tee 14, 36 1.06
Be Se ESCA Bg as OA Se ee ened: ihe TS Se es Oe 45. 52 27.75
It appears, therefore, that the quantity of hydrogen decreased with
the duration of the experiment, while at the same time there was an
266
12 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
increase in the formation of methane, so that in the end the quanti-
ties of carbon dioxid and the methane were about equal. In studying
other substances Popoff found that a methane fermentation could be
produced from gum arabic as well as from cellulose. He is of the
opinion that the typical cellulose ferment gives rise to carbon dioxid
and methane only, and that the presence of hydrogen in the gas is
due to other fermentation processes.
In 1877 Van Tieghem, in working out the life history of the amylo-
bacter of Trecul, found that it was motile, as Nylander had done
twelve years before, and classified it as a bacillus. He further found
that it was an anaerobic, cellulose-dissolving organism and that it
grew readily in soluble starch and cellulose, first reducing them to
dextrin and then converting the dextrin into glucose, which was fer-
mented with the production of carbon dioxid, hydrogen, and an acid
which inhibited the growth of the organism unless neutralized with
calcium carbonate. No cytase was liberated in solution and the
cellulose was dissolved only when in direct contact with the organ-
ism. He proved the cellulose-dissolving power of the organism to
his own satisfaction by inoculation experiments in solutions con-
taining macerated radish. However, he found that the organism did
not act the same on all plant tissues; in a word, that Bacillus amylo-
bacter could not attack all celluloses.
Results of studies on fermentation processes were published by
Prazmowskiin 1880. He described two species to which he attributed
cellulose-fermenting properties and to which he gave the names
Olostridium polymyza and Vibrio rugula. The former was found to
have only a weak fermentive power in dextrin solutions, but was
extremely active in preparations of cooked potato and lupine seed;
its activity on starch and cellulose is described as very vigorous. An
analysis of the gas formed showed only hydrogen and carbon dioxid.
Vibrio rugula is of especial interest because the description given
is so similar to that given later by Omelianski for his so-called hydro-
gen and methane ferments. In young cultures the rods were unusu-
ally thin, about 8 microns long, and showed a characteristic curved
structure which made it easy to separate them from other species;
later the rods became uniformly thicker, the end swelled up, and a
round spore appeared. The young rods were actively motile and
the organism was classed as an anaerobe. In infusions of plant
tissue the organism was found to surround the cell walls, which were
soon dissolved. Prazmowski also made a study of an organism which
stained blue with iodin; to this he gave the name Clostridium butyri-
cum, although he regarded it as synonymous with Vibrion butyrique
Pasteur, Amylobacter Trecul, Bacillus amylobacter Van Tieghem, and
Bacterium navicula Reinke and Berthold.
266
HISTORICAL REVIEW OF INVESTIGATIONS. 13
In an éxtensive series of experiments inaugurated in 1880 Tap-
peiner has given us interesting data on the disappearance of cellulose
in the digestive tract of-herbivorous animals. Incidentally he made
a study of the compounds produced by cellulose fermentation. Pure
cellulose in the form of cotton and filter paper was placed in flasks
containing a rich nitrogenous solution. In one series of experiments
a 1 per cent neutral flesh extract was poured into flasks; pure cellu-
lose in the form of cotton was added, sterilized, and inoculated with
a drop of material from the stomach. It was observed that the
flesh-extract solution invariably resulted in a fermentation inde-
pendent of the typical cellulose ferment; therefore, a check flask
containing only the flesh extract was held under the same conditions
as the flesh-extract cellulose flask. The result of one such experi-
ment is shown in Table IT.
TABLE IT.— Measurement of gas formed by the decomposition of cellulose and flesh
extract and of flesh extract alone.
e-p i _—*
. cellulose es
Gas formed. and flesh | extract
extract. (check).
Ofe3 Ce
oe EO SS 2 A Se ee a er a ek ae 191. 00 10.10
REESE SEES ¢ nS ee ee eee eee es oe. eee es 2 1.70 3.00
EE a ee eee oe ee ee ee»: 10. 40 8. 60
AGM Eee St OR ee, See see ek ook rate JS Rs 88. 30 4. 20
The fatty acids found in flask 1 amounted to 1.6651 grams and con-
sisted of 2.2 parts acetic acid to 1 part butyric acid, giving a carbon
content of 0.7414 gram: The fatty acids found in flask 2 amounted
to 1.005 grams and consisted of 2.1 parts acetic acid to 1 part butyric
acid, giving a carbon content of 0.4918 gram. Subtracting the
products found in flask 2 from those in flask 1 we have the products
formed in the decomposition of cellulose, with a carbon content as
follows: Carbon dioxid, 0.0966 gram; methane, 0.0436 gram; acids,
0.2496 gram. Total, 0.3898 gram. |
In the fermentation of the cellulose 0.4165 gram of carbon was lost,
being slightly more than the quantity recovered in the by-products.
A separate series of experiments was conducted with Nageli’s
solution ‘ and 0.6 per cent asparagin. Flasks of 360 cubic-centimeter
capacity were filled with this solution and 3.5107 grams of dry cotton
added. The fermentation resulted in the formation of carbon dioxid,
hydrogen, and nitrogen. Fatty acids, including acetic, propionic, and
butyric, were also produced. In the control flask no gas was formed
and only traces of fatty acids. It is noted in this experiment that
1 Potassium phosphate (dibasic), 0.20 gram; magnesium sulphate, 0.04 gram; calcium chlorid, 0.02 gram;
water, 100.00 c. c.
266
14 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
0.9447 gram of cellulose was destroyed without any methane for-
mation, while in the previous experiment a considerable quantity
of methane was produced even in the flesh-extract flask to which no
cellulose was added.
The interesting series of experiments commenced by Hoppe-Seyler
in 1881 did much to confirm the growing belief that cellulose is de-
composed by a methane ferment. The main experiment is as follows:
On December 2, 1881, 25.773 grams of pure filter paper were added
to a 1,101 cubic-centimeter flask inoculated with a small quantity
of slime and sufficient water added to bring the mixture up to about
700 cubic centimeters. The flask was protected from the light by a
double layer of black paper and preparations were made to collect
the gas over quicksilver. The flask was kept in this condition four
years. In the early part of the experiment the gas formation was very
active, toward the end of the year 1883 it became weaker, and in the
second half of the year 1885 was scarcely apparent. The experiment
was discontinued on December 6, 1885. Gas analyses to the number
of 95 were made from time to time, the summary of which showed
3,281 cubic centimeters of carbon dioxid and 2,571 cubic centimeters
of methane. An examination of the contents of the flask at the
end of the experiment showed that 15 grams of cellulose had been
consumed, and as no other appreciable quantity of by-products could
be found the author concluded that cellulose is dissolved according
to the following formula:
(1) The hydration of the cellulose with the formation of a hexose,
C ,H,,0;+H,O=C,H,,0,; and
(2) the destruction of the carbohydrate with the formation of equal quantities of
carbon dioxid and methane,
0,H,,0,=3C0.+3CH,
Hoppe-Seyler says in one part of his work that the formation of
carbon dioxid took place only when he found in his solutions living
bacteria which showed no difference from Bacillus amylobacter of
Van Tieghem, and he is therefore of the opinion that the destruction —
of cellulose was due to the activity of this organism.
Gayon in 1883 and 1884 noted the presence of methane in ferment-
ing manure, and from 1 cubic meter of this material well moistened
with water and held at 35° C. he succeeded in obtaining as much as
100 liters of the gas in 24 hours. This fermentation he attributed
to an extremely small anaerobic organism which was cultivable in
nutrient solutions containing either straw or paper, in which it
attacked the cellulose and liberated carbon dioxid and methane.
The extensive experiments of Deherain in 1883 and 1884 on the
aerobic and anaerobic fermentation of straw and manure showed
that in a pile of manure under natural conditions the gas liberated
266
ee “2
EE eX
HISTORICAL REVIEW OF INVESTIGATIONS. 15
at the bottom of the pile is pure methane and carbon dioxid, while
no methane is produced near the surface unless the manure is wet,
when as much as 10 per cent of the gas produced may be methane.
In hermetically sealed flasks the fermentation soon stopped, but on
opening the flasks and resealing them the fermentation began anew,
though it continued for but a short time. He concluded from this
result that the methane ferment is not a strict anaerobe. Occasionally
a fermentation produced hydrogen and carbon dioxid and gave a
slightly acid reaction, due to the formation of butyric acid, while the
pure methane fermentation was always neutral. A microscopic
examination of the liquid of two such solutions showed in both
cases numerous extremely fine rods, which were almost identical,
and the butyric ferment in addition where hydrogen was produced.
Deherain tried Pasteur’s method of successive cultures, but did not
reach an absolute conviction concerning the differences between the
two organisms. A few drops of a manure infusion inoculated into
dextrin and cane sugar solutions gave hydrogen, while a similar
inoculum in a solution containing paper gave methane. A later
experiment under the same conditions reversed these results in that
9 per cent of gas from the fermentation of cane sugar was found to be
methane, while hydrogen was secured from the decomposition of
paper. This evidence was thought sufficient to show the presence of
two different anaerobic ferments, one hydrogen and the other methane.
It sometimes happened in experiments with straw that an acid fer-
mentation took place and that the dominating gas was methane.
This production of acid he believed to be due to a fermentation of
sugar, producing hydrogen and butyric acid, and that such fermenta-
tion was succeeded by the regular methane fermentation.
The results of a study of the fermentation of manure by Schloessing
in 1889 showed that the anaerobic fermentation was much more
active at 52° than at 42° C., and that methane was the predominating,
if not the only, combustible gas given off. Three years later he and
his son carried on some experiments to determine what part bacteria
play in the aerobic and anaerobic fermentation of manure at different
temperatures. The aerobic work showed that no combustible gas
was produced under these conditions and that the bacteria were very
active up to 72.5° C., but that at 81° C. all action ceased; in an
atmosphere of carbon dioxid he obtained a methane fermentation
at 52° but none at 66° C. Once he obtained methane from cow
manure and hydrogen from horse manure at 58° C. In one experi-
ment 124.4 grams of fresh horse manure containing 76 per cent
moisture were kept in an atmosphere of carbon dioxid for two
months at 52° C., and in that time generated 4,217.5 cubic centi-
meters of carbon dioxid and 4,577.4 cubic centimeters of methane,
266
16 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
which are equivalent to 4.72 grams of carbon. In the first 500 cubic
centimeters of gas produced 15.8 cubic centimeters of hydrogen were
also found. An analysis was made to determine the quantity of each
element in the dry material of 124.4 grams of manure. The results
are shown in Table III.
TaBLE III.—Analysis of 124.4 grams of horse manure (dry).
Hydro- : Nitro- | :
Stage. Carbon. gen. Oxygen. gen. Minerals.
Grams. Grams. Grams. | Grams. | Grams.
Before ‘fermentation .<.2... 67.22 se eee 12.67 1, 653 10.70 0. 453 3.69
Attierdtermentation=.2+.%2 $23 oe eee eee 7.92 A125 7.08 . 392 3.79
Loss (=) OF, £80. (4) asiee tee ak 2 oe eee —4.75 —.528 —3.62 —.061 +.10
The loss of 4.75 grams of carbon corresponds very closely with the
4.72 grams found in the carbon dioxid and methane obtained during
the fermentation and represents 37.5 per cent of the carbon in the
fresh manure.
The methane fermentation of cellulose was likened to the alcoholic
fermentation of sugars by Berthelot in 1889, in that it is determined
by living agents, is accompanied by a fixation of the elements of
water, and has a similar thermochemical mechanism. He repre-
sented the fermentation as taking place so that all the hydrogen
of the water enters into one .of the products (methane) while all
the oxygen goes to form carbon dioxid. The total heat thus liberated
was 41 calories, the products being gaseous.
The effect of alkalinity and aerobic or anaerobic conditions on the
progress of fermentation and mode of decomposition of straw was
investigated by Hebert in 1892. The importance of alkalinity in this
fermentation was tested by adding solutions containing from 5 to 10
per cent of potasstum carbonate, ammonium carbonate, and ammo-
nium phosphate to dry powdered straw of known composition. This
straw suspension was inoculated with several cubic centimeters of
urine and incubated at 55° C. After four days the anaerobic flask
containing 5 per cent of salts had produced the greatest quantity of
gas, which was composed of equal parts of carbon dioxid and hydrogen,
but a week later without any change in conditions this flask began
producing methane. The predominance of either of the carbonates
made no appreciable difference in the rapidity of fermentation, but an
excess of ammonium carbonate gave hydrogen at first and methane
six days later and an excess of potassium carbonate gave methane in
the beginning. The composition of the straw before and after three
months’ fermentation is given in Table IV.
266
“I
HISTORICAL REVIEW OF INVESTIGATIONS. 1
TasBLe 1V.—Composition of straw before and after fermentation.
SSNS —
| Composition of straw
after 3 months’ fer-
}
| Initial
mentation.
Constituent. IGCtO SO Ee
of'straw. Anaerobic | Aerobic
fermenta- | fermenta-
tion. tion.
Per cent. Per cent. Per cent.
pith NG) aa NG RD SIRES sah © SE 8 SMa pet J POPES De ie Coes aes eee ss Cee eee Pee ee ee
Berenouseapenialst) 3s i45. SeE0! 22 L ee a esc OA. 358 4.87 12.50 14. 06
MERINGUE IMTTINUCRIANS | hls ap ce srcig osteo ta spon bitte ats Cae oe Sate hehe ett 93 1.36 . 62
OO UCIT 0 EG ENS FSS eS es ee oe eT ee a 2. 43 . 00 . 00
so ST ETLELS yeh ass See RE Se ep ASE Sc SI Soe oooh ie aed - 05 00 . 00
SRT emnIPMe DATUNTIS HCTOS 2 Stee i a 2 a4 bo eet. Sees erty ide o's o's es . 60 1.26 1.60
I 5 APS 0 SI dae nia OG ew he A ae AER ine ke > inte o 28 af 28. 25 23.30 25. 65
MeereiGsee. | ase pee SEO). A) EEA SEA ee) REE). thas. . SEALS 28. 03 22.70 28. 25
Merry (1MLO ey LOB). 2 rls aks Ld. oe ed eh ae et eee | 20. 00 26. 00 19. 00
Reem men ene bE Reg ee SD Es oo a ae at enhela s aed 7.15 12.80 12. 80
Bie Gene CPE Liste eee eee ECT ODS, SE 25h taco Soe ab | 101. 83 99.92 101. 98
Gram. | Gram Gram.
RAHN SULA Wis hs ye Jc SLE hs ode ee rg Se a hae e ool sooo 0. 4524 0. 2645 0. 2690
MveiPhtior cellulose in the straw... 1.000...) 2 fio f sis se3. . 1412 | 0615 . 0689
- The results show that in both fermentations the straw lost about
half its weight and that this loss occurred principally in the three
elements most abundant in straw, namely, cellulose, vasculose, and
straw gum. The straw first lost all or part of the substances most
easily attacked, as chlorophyll materials, gums, tannins, glucose, and
dextrin, the higher carbohydrates, cellulose, and straw gum disap-
pearing afterwards; the loss of the former amounting to as much as
50 per cent (7 or 8 grams). Finally the vasculose was partly dis-
solved in the solution and partly oxidized to carbon dioxid and water.
The organisms appeared to work as vigorously under aerobic as
under anaeorbic conditions, and Hebert was unable to decide from
his figures what are the most favorable conditions for the destruction
of cellulose.
Van Senus in 1890 published an extensive treatise on the decom-
position of cellulose. He noted the rapid decomposition of cotton,
pieces of bean, potato, etc., when moculated with slime and held
under favorable conditions, and attributed these decomposition pro-
cesses to the joint activity of two organisms. One of them, Bacillus
amylobacter, which he describes as a rod-shaped organism, 0.8 “ wide
and 2 to 10 » long, stains blue with iodine and forms spores when air
is admitted, which then germinate only when air is excluded. He
showed that B. amiylobacter in flesh-extract solution with cellulose
(cotton, paper, crude fiber, etc.) under no conditions could ferment
the cellulose; with sterilized beans, potatoes, etc., the walls were not
broken down but only separated from each other, probably through
the formation of the ferment pectase. The other organism, isolated
from the intestines of the rabbit, is much smaller and, like B. amyio-
62420°—Bull. 266—13——3
18 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
bacter, has no power to ferment cellulose in pure culture; however,
when these organisms were grown in association a destruction of cel-
lulose was secured. The reason for this result, in the opinion of this
author, is the production of harmful products by the fermentation of
the cellulose, and the presence of another organism is necessary to
render these products less injurious. Van Senus is of the opimion
that methane is not formed directly by the fermentation of cellulose
and that the destruction of cellulose always results in the formation
of hydrogen, carbon dioxid, and acetic acid, the action of the hydro-
gen upon the acetic acid reducing it to aldehyde, alcohol, and finally
to methane.
In 1894 Omelianski began an investigation of bacterial cellulose
destruction by inoculating a nutrient solution containing filter paper
with slime from the River Neva, and incubating at 30° to 35°C. The
paper was soon changed into a yellowish, transparent, gelatinous
mass, which later disappeared, leaving only a slight residue. A
long, extremely slender bacillus with a round polar spore was present
in great numbers. In order to get a pure culture of this cellulose-
fermenting organism, Omelianski employed Winogradski’s ‘‘method
of elective culture,” selecting for this purpose a nutrient solution
almost void of organic nitrogen and incubating the culture anaerobi-
cally, as the resuits obtained by Hoppe-Seyler had showed that these
conditions were favorable for bringing about a predominance of the
desired organism. After a sufficient number of transfers had been
made this slender rod, Bacillus fermentationis cellulosae, was almost
the only one present in the culture, and when inoculated into solu-
tions containing cotton and cellulose of cabbage, turnip, and pith of
the elder tree which had been precipitated from Schweitzer’s reagent,
it produced a vigorous fermentation with the liberation of hydrogen
and carbon dioxid. A little methane generated in the first cultures
was believed to be due to another organism which disappeared in
transferring. A 300-cubic-centimeter flask was filled with a solution
made as follows:
EARL FMI. cee ats Ye Sn es cine se Sutaess «tans 1.0 gram.
Mipenemui Pakjemeewe.. .. 1S LSC. OL SOLA .5 gram.
ATHMONIDEEAMEDEBTO. . 2 izle lbake ys Ad ee ss {e321 1.0 gram.
Bodin Cee 4. ob do oS 8k a a Trace.
Dictilled-whtat, oa: ...-sieskons scarce ent, deen made ie 1,000.0 ec. e,
To this solution were added 3.4743 grams of dry filter paper and
5.7698 grams of calcium carbonate. It was then inoculated with the
cellulose ferment, purified by the elective culture method, and ineu-
bated 13 months at 35° C. During this time the volume of carbon
dioxid in the total gas increased from 15 per cent at first to 98 per
cent, and toward the end of the period dropped to 80 per cent. At
266
ih
al F
p.
4
HISTORICAL REVIEW OF INVESTIGATIONS. 19
the conclusion of the experiment an analysis of the solution showed
the presence of 2.2402 grams of acetic and butyric acids, m the ratio
of 1.7 to 1, and of gaseous products consisting of 0.9722 gram carbon
dioxid and 0.0138 gram hydrogen, making the total weight of by-
products produced 3.2262 grams. A loss in cellulose, amounting to
3.3471 grams, was noted, being somewhat greater than the total
weights of the by-products found. Valerianic acid, higher alcohol,
products giving the odor of decaying cheese, and dissolved hydrogen
were not measured. These unmeasured products might account for
the difference of 0.1209 gram between the cellulose added and the
products obtained. According to these figures 70 per cent of the
cellulose used is converted into fatty acids, while hardly 30 per cent
is liberated as gas.
In 1904 Omelianski published his method for the separation of the
two cellulose-destroying organisms, one called the ‘“‘hydrogen bacil-
lus,” formerly Bacillus fermentionis cellulosae, and the other the
‘“‘methane bacillus.” The vegetative cells of the methane organism
appeared to form spores more readily than those of the hydrogen
bacillus, so to obtain a pure culture of the latter he first heated the
inoculating material to 75° C. for 15 minutes in order to kill all the
germinating methane organisms, and after several repetitions of this
process his culture was apparently free from this organism. After
five or six generations the surface of the paper was covered with a
bacillus 4 to 8 » long and 0.5 » wide, the rods later reaching a length
of 10 to 15 » without gaining in thickness. They never formed
chains and took the ordinary anilin stains readily, but would not
color blue with iodin. Slightly curved or irregular spiral forms
were ebundant when fermentation was going on, and in older cul-
tures the rods had a round polar spore 1.5 «in diameter. When the
paper had been destroyed there were many free spores and few rods
in the solution. These spores were found to withstand a heat of
90° C. for 25 minutes, so the cultures were freed of nonspore formers
in this way. After such heating there still persisted in his cultures
a large bacillus with an oval polar spore, while another contaminat-
ing organism with a round polar spore and very much like the true
cellulose ferment was occasionally present. The former grew read-
ily on solid media like agar jelly, filter paper soaked in gummy salt
solution, carrots, summer cabbages, turnips, and potatoes, either
alone or in association with other bacteria, but the true hydrogen
ferment did not grow on any of these media except that in one case
some very small, yellow, liquefying, semitransparent colonies of the
cellulose bacteria appeared on a potato plate; they were scarcely
apparent without a magnifying glass, and this medium was evidently
not a favorable one, as a heavy inoculation gave only a few colonies.
266
20 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
Omelianski asks, ‘‘Was the mother culture not absolutely pure or is
the growth an association of bacteria?”’ Inoculation from these
colonies gave fermentation of cellulose in only one case and that
soon stopped. However, the organism from these colonies had all
the morphological characteristics of the bacteria found in the fer-
menting paper, and he concludes that it was without doubt a pure
culture of this bacillus, although it did not give a satisfactory fer-
mentation.
A test to determine whether it was an associative action with the
bacillus forming the oval spore gave negative results. The methane
ferment was obtained by inoculating a flask containing the mineral
salt solution, paper, and chalk with canal slime or fresh horse manure,
and incubating anaerobically at 35° C. A microscopic examination
of the paper showed that it was covered with an organism similar to
the hydrogen bacillus, but thinner and more delicate in outline. The
culture was purified by transferring and heating to kill nonspore form-
ers until it presented a microscopically pure picture, appearing as a
rod 5 » long and 0.4 » wide, with a round polar spore 1 » in diameter.
Morphologically these organisms might be classed as the same species,
but physiologically they were very different, for one produced hy-
drogen and the other methane.
In later investigations Omelianski points out that methane may
be produced not only from cellulose but from acetates, pentoses, pen-
tosans, butyrates, lactates, and protein bodies, which, he believes,
indicates that the number of reactions in nature which involve the
formation of methane is no smaller, perhaps, than the fermentation
processes leading to the evolution of hydrogen.
Experiments showing the destruction of the mner tissue of the
turnip due to the parasitism of Pseudomonas campestris were reported
by Smith in 1902. The leaves of the plant were inoculated with a
pure culture of this bactert'um. The disease moved downward, and
sections of the root 52 days after inoculation showed the bacteria to
be very abundant in the inner part, although the root was entirely
white and sound externally. Cultures made from the diseased inte-
rior yielded only P. campestris. Carefully prepared sections showed
all stages in the solution of the cell walls, from single cells or vessels
occupied by the bacteria to cavities filling the place formerly oceu-
pied by hundreds of cells and filled with the bacteria and remnants
of the cell walls.
Experiments by Van Iterson in 1904 have shown that the fermen-
tation of cellulose may be caused by aerobic as well as by anaerobic
bacteria. According to his results the anaerobic processes fall into
two groups:
(1) Without the presence of nitrates the cellulose may undergo a.hydrogen or
methane fermentation.
266
HISTORICAL REVIEW OF INVESTIGATIONS. 21
(2) In the presence of nitrates the cellulose is destroyed by denitrifying bacteria
according to the following formule:
C,H,.0;+8KN O0,=4KHCO,+2K,CO,+4N.+3H,0.
The destruction of cellulose under aerobic conditions also falls into
two classes:
(1) Ifthe medium isslightly alkaline, certain aerobic bacteria will play the principal
role.
(2) If the medium is acid, then the molds and higher fungi are the active agents of
destruction.
In a simple nutrient solution containing dibasic potassium phos-
phate, potassium nitrate, and filter paper inoculated with a cubic
centimeter of canal water and kept at 35° C., the process started in
6 days, and in about 15 days all of the nitrate added and all of the
nitrite formed in the early stages of fermentation had disappeared.
Analysis of the gas produced showed only carbon dioxid and nitro-
gen, with no trace of hydrogen or methane. Thus, the process here
would seem to be entirely different from the processes resulting in
the production of hydrogen or methane.
For the study of cellulose destruction by aerobic bacteria the fol-
lowing solution was prepared:
SmC EE hr A a 0) on Re eS 100. 00 c. ¢.
ME SU RURE ALES ALL 2 tty. 4s eRe Gl” 2. 00 grams
eA Ia COP tus. vtee - - bie els ry? - alte: © - 4 . 10 gram
Powmesium phosphate (diabasic).......-..-.--,.4tbie--.- . 05 gram.
NMI CON UONALC one Uy oes cle oo a ye eee, Se - 2. 00 grams.
After inoculating with sewer slime the fermentation starts in five
to six days, after which it goes on very rapidly. The fermenting
paper was found to contain a variety of forms, among which was a
very small bacillus frequently associated with a large micrococcus.
The former was believed to be the active agent of fermentation and
was given the name of Bacillus ferruginus. The micrococcus is
described as having no cellulose-dissolving power in itself but as
stimulating B. ferruginus to greater activity when associated with it.
Experiments by Macfadyen and Blaxall with thermophilic bacteria
in 1899 showed that these organisms may be very active destroyers
of cellulose. A nutrient solution containing pure filter paper was
ineculated with a small quantity of soil and incubated at 60° C.
The result was an active development of gas and odor, and in 10 to
14 days the filter paper was completely broken up. The experiments
were repeated with filter paper and also with films of cellulose hydrate
obtained from the solution of cotton fiber in the form of thiocarbonate.
1 Potassium nitrite, potassium nitrate, peptone, or magnesium ammonium phosphate may replace the
ammonium chlorid.
266
22. DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
The following results were reported on the solutions which showed a
disintegration of cellulose by the thermophilic organisms:
(1) No reduction CuO in original or after boiling with acid.
(2) No other evidence of any proximate products of resolution, i. e., carbohydrates
of dimensions NC,.
(3) On distillation 25 c. c. gave volatile acid=1 c. c. of normal NaOH, containing
acetic and butyric acids. Residue gave traces only of furfurol on distillation with
HCl (1.06 s. g.).
It appears that the destruction has been, for the most part, complete, probably to
CO, and CH,. On further investigation you may be able to get an intermediate stage
or an organism acting less severely.
All the above results were brought about by mixtures of thermophilic bacteria
occurring naturally in the soil, and the action appeared to be of a symbiotic nature.
Their action resulted in a complete disintegration of filter paper, fibrous cellulose, and
_ esparto cellulose.
Distaso in 1911 described an organism isolated from the intestinal
flora of the chicken, to which he gave the name Bacillus cellulosae
desagregans, because it was found to be capable of destroying cellu-
lose. When cultivated in mineral solutions with pure cellulose (Ber-
zelius filter paper) the paper is disintegrated, forming flocci or fibers;
action never goes further and this author is not sure whether the
filter paper is thoroughly transformed. The organism does not stain
by Gram’s method; forms oval subterminal spores; is aerobie in
nature; grows in sugar gelatin; never gives off gas; grows well at 37°
but not at 22° C.; produces no indol; grows only feebly on glucose;
does not assimilate lactose, maltose, or saccharose, but transforms
starch into glucose rapidly.
In 1911 Choukevitch in a study on the bacterial flora of the large
intestine of the horse always obtained a fermentation of cellulose in
Omelianski’s nutrient solution when inoculated with several loopfuls of
the intestinal contents. In the fermenting solutions a small organism
(Bacillus gasogenous) was always found, which morphologically
resembled the hydrogen and methane ferments described by Omeli-
anski. Neither a pure culture of this organism nor any of the others
which he isolated from the intestine of the horse was able to ferment
~ cellulose.
The most recent contribution to our knowledge of cellulose destruc-
tion is that of Kellerman and McBeth, whose report is a preliminary
one of work undertaken by the Office of Soil-Bacteriology: Investi-
gations. In this report special attention is given to that portion of
Omelianski’s work from which he concluded that cellulose undergoes
either a hydrogen or methane fermentation. The impurity of Omeli-
anski’s cultures are discussed, and three new species of cellulose fer-
ments isolated from his cultures (Bacillus rossica, B. amylolyticus, and
Bacterium flavigena) are described.
266
i
4
HISTORICAL REVIEW OF INVESTIGATIONS. 23
INVESTIGATIONS WITH FILAMENTOUS FUNGI.
Early experiments with parasitic fungi indicated that many of
these organisms were able to make their way into plant tissues by
piercing the cell membrane. Such observations were made by Kiihn
in his study of blight-producing fungi, by the brothers Tulasne in
their study of the rusts, and by De Bary in studies with Peronospora.
Some years later Marshall Ward was able to watch the penetration
of the cellulose walls of the lily bulb by parasitic fungi. The walls
became swollen and evidently somewhat softened, which condition
he believed to be due to the production of a ferment drop at the tip
of the hyphez. Miyoshi has recently observed a similar phenomenon
with Penicillium glaucum, Botrytis bassiani, and Botrytis tenula. The
power of fungi to destroy cellulose was also early suggested by Hartig
in his studies on the destruction of woody tissues.
Later, the destruction of cellulose by fungi was observed in studies
undertaken primarily to determine the causative agent of some
common plant diseases. In this connection may be mentioned the
work of DeBary with Sclerotina libertiane, Kissling in his biological
studies of Botrytis cinerea, and Behrens with diseases of fruits.
The work of Van Iterson on the destruction of cellulose by fungi
deserves special mention, as it gave the first indication of the extent
of cellulose destruction by fungi. He was also the first to devise a
method for isolating these organisms. To that purpose two sterile
sheets of pure filter paper were placed in a Petri dish and moistened
with the following solution:
Beem uaa Bit ot be Bio eon in eee, Pees asks og 100. 00 c. ¢.
mimmomum nitrate js. - 2-5 - --.)- »- RE 1 tt. ee . 05 gram.
folassium phosphate (monobasic). . 2... /...--...---.---.- . 50 gram.
For inoculating material, soil or humus was used; however, the
best results were obtained by exposing the dish to the air 12 hours
and then cultivating at 24° C., taking care that the paper remained
moist. After two or three weeks the paper was covered by a rich
mold growth including a large number of species; among them were
several species seldom found on malt gelatin, and on further study
several of these were found to be active cellulose destroyers. The
great abundance of these mold spores in the atmosphere was shown
by the following experiment.
A Petri dish having a surface of 275 centimeters and containing
filter paper moistened with the solution previously described was
allowed to stand open in the garden 12 hours; 152 mold colonies
developed, comprising 35 different species. It is evident from this
experiment that large numbers of cellulose-destroying mold spores
were floating in the air. The fungi found growing upon the paper
were purified by means of malt gelatin. The destruction of cellulose
266
24 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
by pure cultures was then studied by inoculating sterile nutrient
solutions containing pure filter paper prepared as previously described.
In this work the following 15 species were isolated and described:
Sordaria humicola Oud., Pyronema confluens Tul., Chaetomium
Kunzeanum Zopf., Pyrenochaeta humicola Oud., Chaetomella horrida
Oud., Trichocladium asperum Harz., Stachybotrys alternans Oud.,
Sporotrichum bombycinum (Corda) Rabh., Sporotrichum roseolum
Oud. and Beijer., Sporotrichum griseolum Oud., Botrytis vulgaris Fr.,
Mycogone puccinioides (Preuss) Sacc., Stemphylium macrosporoideum
(B. en Br.) Sacc., Cladosporium herbarum (Pers.) Link., and Epicco-
cum purpurascens Ebrenb.
Three years later the work of Appel appeared, which showed that
certain forms of Fusarium can destroy cellulose with great rapidity.
Ten grams of pure, dry filter paper were introduced into an Erlen-
meyer flask, and 50 cubic centimeters of a nutrient solution contain-
ing potassium phosphate, magnesium sulphate, and potassium
nitrate were added. An intensive cellulose fermentation developed,
resulting in the destruction of 80 per cent of the paper in 14 days.
Since all species of Fusarium can not use cellulose as a source of
energy, the writer believes this fact can be made a valuable pomt in
identifying the species of Fusarium. :
In a series of experiments with a comparatively large number of
molds Schellenberg showed that the destruction of the hemicellulose
in plant tissues is sharply separated from the destruction of real
cellulose, and also that molds act differently toward the hemicellulose
of different plants. This difference in action is thought to be due to
the differences in chemical composition of the plant tissues rather
than to differences in solubility of the hemicelluloses. The results
of this work are summarized in Table VY.
TaBLeE V.—-Action of molds on real celluloses and hemicelluloses from various plants.
| Cellulose
from H a from—
Name of plant. ———_—_—__— eee es ae *
Mo- Lupi- tas Phoe- | Impa-
Cotton.| Hemp.) jinia. | ‘nus. nix. | tiens.
/
|
|
Mucor TacemMosus J. cic Eee «3 0k Ra |
AIUCOr DOPIDOUUIS..S.2 nwa cen caee aie - ++ sescsnbe
Mucor pirtiorme si: «Ac pce wees oss 160 bee
MUCOI ZIGDOSIIS: ..cnctc cpueaeeRUnee << =0 06s ster o .
Thammeidium elevans.isiksvsakee- - «> swhxas
FRIZO0US MICTICANG: 2-5. t.ccawee@e- ss cos sacene
Penicillium glaucum 5 Ck a apy /
Penicillium glaucum
Sclerotinia fructigena...........0.-----.--+++:!
Sclerotinia Cinerea.......Sacchwcwaeccesceacecs
Botrytis cinerea... .......seeeeeeee---- see sees]
BOUryuis VIGArIS. |... ck cwms Semen s~ secs ence -
Nectria cinnabarina. ............----..+--++5- _
Cladospouitinn Herbarium: to ewenss- -¢%. 20-505.
Colletotrichum lindemuthianum..............
‘Trichatsieelum TOSCUMIL Ie «dead =. .- chs occas]
wee ee mete eee eee eee eee!
per kat. Lae | ke bet
’
terre, eb Pear ht ot
$++H+t4+G+ 1444441
HVPET PPt bei rad
HEEL EEL Ltt tttet |
a
ST eta oe te tet Ae USF
og Pe el it 8 Pw A ae, a Pe
266
——
METHODS AND CULTURE MEDIA EMPLOYED. 25
The recent work of Marshall Ward suggests the importance of
Penicillium as a wood-destroying fungus. Spores of a pure culture
of Penicillium were sown on sterile blocks of spruce wood; the mold
grew freely and developed large quantities of spores on normal
conidiophores. Sections of the wood showed that the hyphe had
entered the starch-bearing cells of the medullary rays of the sap-
wood and consumed the whole of the starch. In cultures three
months old the hyphe were found deep in the woody tissue passing
from tracheid to tracheid via the border pits. In conclusion Ward
says:
Tt certainly looks as if Penicillium may be a much more active organism in initiating
and carrying on the destruction of wood than has hitherto been supposed, and that it
is not merely a hanger-on or follower of more powerful wood-destroying fungi.
Bourquelot has shown the great versatility of Aspergillus in the
production of enzymes, having found it capable of producing invertase,
maltase, trehalase, emulsin, inulase, diastase, and trypsin, and
Bertrand and Holderer have found that it also produces cellase.
Ward suggests that Penicillium may be equally rich in the capacity
for enzyme production.
Among the higher fungi Schornstein found that Pora vaillanti,
Polyporus vaporarius, Polyporus destructor, Coniophora cerebella,
and Paxillus panuoides are capable of destroying wood, which, as is
well known, is largely composed of cellulose. Polyporus destructor
quickly forms fruiting bodies and never entirely destroys the wood.
Murulius lacrymans and M. pulverulentus appeared on wood soon
after it had been built into position and entirely destroyed it.
Arzberger in an investigation of the fungus which causes root
tubercles on Ceanothus and Eleagnus found that it belonged to the
genus Frankia and secreted an enzyme that destroyed the cell walls.
METHODS AND CULTURE MEDIA EMPLOYED.
In taking up the study of cellulose fermentation the elective
cultural method employed by Omelianski and the method of Van
Iterson of using sheets of filter paper were tried under both aerobic
and anaerobic conditions. Microscopic examinations of the cultures
kept under anaerobic conditions showed the presence of organisms
similar to the hydrogen and methane ferments of Omelianski. In
young cultures these organisms appeared only in small numbers,
but became very numerous as the decomposition process advanced.
In cultures kept until the paper had been completely destroyed, the
spores of this organism became extremely numerous and microscopi-
cally the cultures appeared to bealmost pure, but the presence of many
other species was easily demonstrated by plating on ordinary media.
The cultures kept under aerobic conditions showed no organisms
62420°—Bull. 266—13——4
\
26 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
resembling the hydrogen or methane ferments which were so numer-
ous in the anaerobic cultures, although the inoculum used was the
same. The decomposition of the paper, however, was most rapid
in the aerobic flasks. This was in accordance with Van Iterson’s
observations that cellulose may be rapidly decomposed by aerobic
organisms. Plates from these cultures, like those grown anaerobi-
cally, showed the presence of several species of bacteria, even after
numerous transfers. Our failure with these methods, together
with Omelianski’s admission of the impurity of his cultures after
the most painstaking care to purify them, led us to believe that no
accurate knowledge of cellulose fermentation could be obtained until
a satisfactory plating medium had been secured.
In taking up the question of a suitable medium, a large number of
nutrient solutions were first studied, including beef broth, decoctions
of plant tissues, soil extracts, manure extracts, and numerous syn-
thetic solutions. The following solution was finally adopted as giving
the best results:
Potassium phosphate (dibasic).............--.-------- 1 gram.
Magnesium: sulphate.\. .'. 2. 5. 3.52) 015 a aes ee 1 gram.
Soditimr catbongiey: +6. 4+). nen dob Eb eet eee one 1 gram.
Ammontanmi pilphate, s\...- 2 copied ieee 7. Plate cultures—
| Cellulose agar, 15 days.
Form: Surface and bottom, round; embedded, lenticular or irregular.
Size: Surface and bottom, 1 to 4 mm.; embedded 0.7 to 1.8 mm. on
major axis.
Enzymic zone: 0.3 to 1.6 mm.
Elevation: Raised.
Topography: Smooth.
Consistency: Slimy.
Chromogenesis: Surface and bottom, reflected light, gray with yellowish
or whitish nucleus; transmitted light, brown. At angle of 45° by
transmitted light bottom colonies show interior of colony bluish or
iridescent and white ring around border. Embedded, reflected
light, white; transmitted light, opaque.
Internal structure: Surface and bottom granular with opaque to trans-
lucent granular nucleus, and frequently having finely granular ring
at border; embedded, opaque, often with numerous outgrowths. .
Edge: Entire to undulate.
Potato agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular to triangular.
Size: Surface and bottom, 1 to 6 mm.; embedded, 0.5 to 1.5 mm.
Elevation: Convex.
Topography: Smooth.
Consistency: Slimy.
Chromogenesis: Surface, reflected light, glistening grayish white; trans-
mitted light, opaque or translucent brownish gray, often with
opaque nucleus. Embedded, reflected light, white; transmitted
light, opaque. Bottom, reflected light, gray; transmitted light,
light brown, at 45° bluish and often iridescent.
Internal structure: Surface, homogenous, opaque or finely granular,
often with lenticular or round nucleus; embedded, opaque, some-
times with translucent irregular, finely granular outgrowths; bottom,
homogenous, finely granular, frequently with granular nucleus.
Edge: Entire. ;
Beef agar, 5 days. |
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, 1 to 4 mm.; embedded, 0.6 to 1.5 mm. on
major axis.
Elevation: Convex.
Topography: Smooth.
Consistency: Slimy.
Chromogenesis: surface and bottom, reflected light, yellowish or grayish
white; transmitted light, translucent brown; at 45° iridescent ring
at border. Embedded, reflected light, yellowish or grayish white;
transmitted light, opaque.
Odor: None.
266
lon)
32 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
II. Cultural features—Continued.
7. Plate cultures—Continued.
Beef agar, 5 days—Continued.
Internal structure: Surface, granular, often with lenticular nucleus and
finely granular and hyaline at edge; embedded, opaque; bottom,
granular, sometimes with nucleus.
Edge: Entire.
Starch agar, 5 days.
Form: No surface colonies; embedded, lenticular to irregularly round;
bottom, round.
Size: Embedded, 0.4 to 1.2 mm.; bottom, 1 to 1.5 mm.
Enzymic zone: 1.7 to 3 mm.
Elevation: No surface growth, but agar raised by colonies just below
surface.
Chromogenesis: Embedded, reflected light, white; transmitted light,
opaque. Bottom, reflected light, opalescent or white; transmitted
light, barely translucent, dark gray.
Internal structure: Embedded, opaque and often irregular with out-
growths; bottom, granular, generally becoming finely granular at
edge, and usually with lenticular nucleus.
Edge: Entire.
Dextrose agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, 0.8 to 1.2 mm.; embedded, 0.4 to 1 mm.
‘Elevation: Convex.
Topography: Smooth.
Consistency: Slimy.
Chromogenesis: Surface, reflected light, grayish white, generally with
white nucleus; transmitted light, translucent brownish gray. ‘ Em_
bedded, reflected light, white; transmitted light, opaque. Bottom,
reflected light, gray; transmitted light, smoky brown.
Internal structure: Surface and bottom, finely granular, usually with
round or lenticular nucleus; embedded, opaque, granular at edge.
Edge: Entire.
‘TIT. Physical and biochemical features.
1. Peptone water—
Addition to peptone water.
Test for— Dex- | Lac- |Saccha-] Malt- | Glye- | Mam- | goin
trose. | tose. rose. ose. erin. nite. 3
Gas DNOGURTION ie is uso deen ob ken on 0 0 0 0 0 0 0
Acid production, 6 days.....-........-. 1. 42 -70 1. 60 1.45 . 53 0 1.48
1.63 1.43 . 76 1. 62
Acid production, 12 days............... 1. 60 . 90
2. Dunham’s: Ammonia trace.
3. Nitrate broth: Nitrite +; ammonia —.
4. Peptone-nitrite solution: Indol +-.
BACTERIUM LIQUATUM, N. SP.
: (Pl. I, figs. 1, 2, and 3.)
I. Morphology.
1. Vegetative cells from 24-hour cultures at 30° C. Beef agar, average length
1.7 #, maximum length 2.6 », width 0.4». Potato agar, average length 0.8
#, Maximum length 1.5 », width 0.3 p.
. No endospores.
3. Staining reactions: Methylene blue +: carbol fuchsin +; Gram —.
266
2 bo
Aettee me
Po -<
”
4
ry
4
.
DESCRIPTIONS OF CELLULOSE-DESTROYING BACTERIA. 33
If. Cultural features.
1. Agar strokes, 10 days. General characteristics: Glistening, smooth, moist
os)
co OH.
growths.
Beef agar: Abundant, raised, grayish yellow; agar whitened.
Potato agar: Abundant, flat, watery, pale grayish yellow.
Dextrose agar: Scant, flat, watery, vitreous to pale yellow.
Starch agar: Moderate, flat, vitreous to pale yellow.
Cellulose agar: Moderate, convex, vitreous to pale yellow.
. Potato: After 15 days growth abundant, glistening, smooth, brilliant canary
yellow; potato not colored.
. Agar stab: Growth best at top, papillate.
. Gelatin stab: After 15 days liquefaction napiform; after 50 days stratiform
with liquefied gelatin present.
. Beef broth: Moderate clouding, scant compact sediment.
. Litmus milk: Faintly acid in two days.
. Plate cultures—
Cellulose agar, 15 days.
Form: Surface and bottom, round; embedded, lenticular to irregularly
round.
Size: Surface, 1 mm.; embedded, 1 mm. on major axis; bottom, 1.5 mm.
Enzymic zone: 0.4 to 0.75 mm. wide and slightly depressed.
Elevation: Raised or umbilicate. Embedded colonies just below
surface give umbonate appearance due to depression of enzymic
zone. ,
Topography: Smooth.
Consistency: Soft.
Chromogenesis: Surface, embedded and bottom, reflected light, faint
yellowish gray; surface and bottom, transmitted light, translucent
gray generally with opaque nucleus, surface sometimes showing
opaque ring at edge; embedded, opaque.
Internal structure: Surface and bottom, granular, generally with round
or lenticular nucleus; embedded, opaque, sometimes conglomerate.
Edge: Surface and bottom, irregular, finely granular; embedded, entire.
Starch agar, 5 days.
Form: Round.
Size: Embedded and bottom, 0.5 to 2 mm.
Enzymic zone: 1 mm.
Elevation: No surface colonies. Medium may be raised by embedded
colonies just below surface.
Chromogenesis: Embedded, reflected light, white, opaque. Bottom,
reflected light, light gray with whitish gray nucleus; transmitted
_ light, gray with opaque nucleus or like embedded.
Internal structure: Embedded and bottom, central area opaque, becom-
ing coarsely granular near edge.
Edge: Embedded, irregular, granular; bottom, blending with medium
or irregularly granular.
Beef agar, 5 days.
266
Form: Surface and bottom, round; embedded, lenticular to round.
Size: Surface, 1.5 to 3 mm.; embedded, 0.5 to 1 mm.; bottom, 1 to
1.5mm.
Elevation: Convex to pulvinate.
Topography: Smooth.
Consistency: Slightly viscid.
34 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
Il. Cultural features—Continued.
7. Plate cultures—Continued.
Beef agar, 5 days—Continued.
Chromogenesis: Surface, reflected light, glistening, sebaceous; trans-
mitted light, translucent brown, some with vitreous edge. Embed-
ded, reflected light, sebaceous; transmitted light, opaque. Bottom,
reflected light, gray; transmitted light, translucent brown.
Odor: None.
Internal structure: Surface, coarsely granular, generally showing irregu-
lar granular nucleus; embedded, opaque, often showing outgrowths
in all directions; bottom, granular, with a granular nucleus.
Edge: Entire.
Potato agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular...
Size: Surface, 1 to 5 mm.; embedded, 0.5 to 1.5 mm. on major axis;
bottom, 1 mm., sometimes spreading to 15 mm.
_ Elevation: Convex.
Topography: Smooth.
Consistency: Watery.
Chromogenesis: Surface, reflected light, glistening, opalescent; trans-
mitted light, edge bluish and iridescent. Embedded, reflected
light, cream color; transmitted light, opaque. Bottom, reflected
light, gray; transmitted light, vitreous, sometimes with a brownish
central area.
Internal structure: Surface, granular, often almost hyaline, generally
showing nucleus; embedded, granular, sometimes showing nu-
merous outgrowths; bottom, finely granular, generally with granular
nucleus and often having grumose center.
Edge: Entire.
Dextrose agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, | to 2 mm.; embedded, 0.5 to 0.75 mm.
Elevation: Convex.
Topography: Smooth.
Consistency: Watery.
Chromogenesis: Surface, reflected light, vitreous to gray, with whitish
gray central area often showing white nucleus; transmitted light,
translucent light brownish gray, with vitreous edge and opaque
nucleus. Embedded, reflected light. grayish white; transmitted
light, opaque; bottom, like surface, no nucleus.
Internal structure: Surface, finely granular, usually with opaque len-
ticular nucleus; embedded, opaque, but showing granular at edge;
bottom, finely granular with small granular nucleus.
Edge: Entire. |
IIL. Physical and biochemical features.
|. Peptone water—-
Addition to peptone water.
Test for— / | am 9 Mi
Dex- | Lac- ‘Saccha- Malt- | Glye- =
trose. ) tose. I Ses ose. erin. ~ | Starch
|
ee L NI | ies ey f ieee ee
Gas production... cipal 2. v.540%. 3. 5% 0 0 ie 0 0 0
Acid production, 6 days................ 1. 20 | 50 1.32 1.16 08 0 1.38
Acid production, 12 days.............. 1.27 8 | 1.33} 1.16 .3| 0 | 1. 40
266
DESCRIPTIONS OF CELLULOSE-DESTROYING BACTERIA. 35
Il. Physical and biochemical features—Continued.
2. Dunham’s: Ammonia +.
3. Nitrate broth: Nitrites +; ammonia —.
4. Peptone-nitrite solution: Indol +.
BACILLUS BIBULUS, N. SP.
(Pl. II, figs. 4, 5, and 6, and Pl. IV, figs. 1 and 2.)
I. Morphology.
1. Vegtative cells from 24-hour cultures at 30° C. Beef agar, average length 1.3 p,
maximum length 2.0 », width 0.4 ». Dextrose agar, average length 0.8 4,
maximum length 1.4 », width 0.4 yp.
2. No endospores.
3. Staining reactions: Methylene blue +; carbol fuchsin +; Gram —.
4 Il. Cultural features.
a 1. Agar strokes, 10 days. General characteristics: Glistening, smooth, moist,
5 raised or convex growth.
i Beef agar: Abundant, grayish yellow.
i Potato agar: Abundant, grayish yellow.
i Dextrose agar: Scant, white.
4 Starch agar: Scant, vitreous to gray.
R Cellulose agar: Moderate, yellowish.
2. Potato: After 15 days’ growth abundant, glistening, smooth, brilliant canary
yellow. Potato not colored.
3. Agar stab: Growth best at top, papillate.
4. Gelatin stab: After 15 days’ line of puncture filiform, later echinulate; lique-
faction crateriform. After 50 days deeply crateriform, no liquefied gelatin
present.
5. Beef broth: Slight clouding, scant compact sediment.
6. Litmus milk: Faintly acid in two days.
7. Plate cultures—
Cellulose agar, 15 days.
Form: Surface and bottom, round; embedded, round to irregularly round.
Size: Surface and bottom, 0.5 to 0.8 mm.; embedded, 0.3 to 0.5 mm.
. Enzymic zone: 0.3 mm. in some cases.
Elevation: Convex.
Topography: Smooth.
Consistency: Soft.
Chromogenesis: Surface and bottom, reflected and transmitted light,
opalescent, usually with grayish white opaque nucleus. Em-
bedded, reflected light, grayish or yellowish white; transmitted
light, opaque.
Internal structure: Surface granular, sometimes with clouded radial
areas extending to edge of colony or with a grumose center; em-
bedded, granular, may show lenticular mother growth with numerous
outgrowths; bottom, granular, often with roundish granular nucleus,
and may also be clouded.
Edge: Irregular and granular.
Starch agar, 5 days.
Form: No surface growth; embedded, irregularly round to round;
bottom, round.
Size: Embedded and bottom, 0.3 to 2.5 mm.
Enzymic zone: 1 to 2.56 mm.
a Elevation: Medium slightly raised by colonies just under the surface.
sh _Chromogenesis: Embedded and bottom, reflected light, white; trans-
mitted light, translucent gray to opaque.
266
36 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
Il. Cultural features—Continued.
7. Plate cultures—Continued.
Starch agar, 5 days—Continued.
Internal structure: Embedded, narrow finely granular zone around
edge, remainder opaque; bottom, finely granular zone at edge
wider than embedded; remainder opaque.
Edge: Embedded, entire to irregularly granular; bottom, blending
with medium.
Dextrose agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and embedded, 0.4 to 0.6 mm.; bottom, 0.5 to 1 mm.
Elevation: Convex.
Topography: Smooth.
Consistency: Simy.
Chromogenesis: Surface, reflected light, white to faint yellowish gray;
' transmitted light, barely translucent, dark brown. Embedded,
reflected light, like surface; transmitted light, opaque; bottom,
reflected light, opalescent.
Internal structure: Surface, finely granular, with an opaque round or
granular nucleus; embedded, opaque; bottom, finely granular,
sometimes with nucleus. .
Edge: Entire.
Beef agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, 1 to 2mm.; embedded, 0.5 to 1 mm. on major
axis,
Elevation: Convex.
Topography: Smooth.
Consistency: Soft.
Chromogenesis: Surface, reflected light, light yellow; transmitted light,
translucent brownish yellow, may have opaque nucleus. Embedded
reflected light, yellow to yellowish gray; transmitted light, opaque.
Bottom, reflected light, gray; transmitted light, translucent smoky
brown.
Internal structure: Surface, granular, with dark round or lenticular
nucleus, sometimes with hyalin edge; embedded, granular, occa-
sionally with outgrowths in one or two planes; bottom, granular,
usually with small granular nucleus, and often with edgelike surface.
Odor: None.
Edge: Entire.
Potato agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, 0.5 to 1.5 mm.; embedded, 0.5 to 0.8 mm. on
major axis.
Elevation: Pulvinate.
Topography: Smooth.
Consistency: Soft.
Chromogenesis: Surface, reflected light, yellowish gray; transmitted
light, barely translucent brown. Embedded, reflected light, yel-
low; translucent light, opaque. Bottom, reflected light, gray;
transmitted light, translucent brown.
Internal structure: Surface, finely granular, generally with round or
lenticular opaque or granular nucleus, often with hyalin edge;
embedded, opaque; bottom, like surface except nucleus, which if
present is small, round, and granular.
Edge: Entire.
266
DESCRIPTIONS OF CELLULOSE-DESTROYING BACTERIA. 37
/
Ill. Physical and biochemical features.
1. Peptone water—
Addition to peptone water.
Test for—
Dex- Laec- | Saccha-| Malt- | Glyc- | Man-
trose. tose. rose. ose... par ag nite. Starch.
GAS PLOGUCHION . 2... 2.05240. fee 0 0 0 0 0 0 )
Acid production, 6 days......... 1.75 1,20 1.57 1.22 15 -75 1.90
Acid production, 12 days........ 1.85 1. 28 1.50 1. 47 ao 1.20 2. 07
i. 2. Dunham’s: Ammonia +.
3. Nitrate broth: Nitrites —; ammonia —.
4. Peptone-nitrite solution: Indol, trace.
, ; PSEUDOMONAS SUBCRETUS, N. SP.
: (Pl. I, figs. 4, 5, and 6.)
I. Morphology.
: 1. Vegetative cells: Starch agar, 24 hoursat 30° C., average length 1.2». Maximum
length 1.4 », width 0.3 ». Beef agar, 48 hours at 30° C. (no growth in 24
’ hours), average length 1.4 », maximum length 3.0 », width 0.4 p.
2. No endospores.
3. Staining reactions: Methylene blue +; carbol fuchsin +; gram —,
Il. Cultural features.
1. Agar strokes, 10 days. General characteristics; glistening, smooth, moist, |
vitreous to faint yellow.
Beef agar: Moderate, flat.
Potato agar: No growth.
Dextrose sugar: Scant.
Starch agar: Moderate.
Cellulose agar: No surface growth. Moderate, generally faint yellow growth
in medium, area of growth sunken.
2. Potato: After 15 days’ growth scanty, concave due to slight liquefaction of
potato, white to faint yellow. Potato bleached around growth.
. Agar stab: Growth best at top, papillate.
. Gelatin stab: After 15 days filiform, no liquefaction.
. Beef broth: No growth.
. Litmus milk: No growth.
. Plate cultures—
Cellulose agar, 15 days.
Form: Round.
Size: Average 3 mm.
Enzymic zone: Enzyme acts within the colony; older colonies may show
narrow clear zone about 0.3 mm. wide.
Elevation: Medium concave, no surface growth.
Chromogenesis: Reflected light, transparent gray or yellowish gray:
transmitted hght, smoky brown.
Internal structure: Central area clouded light brown, then a zone like the
medium but less dense, sometimes surrounded by a denser border line
which may be broken.
Edge: Entire to undulate.
Starch agar, 5 days.
Form: Round.
Size: Surface and bottom, 0.3 to 1.5 mm.; embedded, 0.5 to 0.7 mm.
Enzymic zone: 2 to 4mm.
Elevation: Convex.
; 266
“NJ ote» &
}
‘
38 DESTRUCTION OF CELLULOSE BY BACTERIA AND FUNGI.
Il. Cultural features—Continued.
7. Plate cultures—Continued.
Starch agar, 5 days—Continued.
Topography: Smooth.
Consistency: Very soft.
Chromogenesis: Surface, reflected light, yellowish white; transmitted
light, opaque nucleus surrounded by narrow or wide translucent
yellow zone. Embedded, reflected light, yellowish white; trans-
mitted light, opaque. Bottom, reflected light, gray with grayish
white nucleus; transmitted light, translucent yellowish gray with
opaque nucleus. |
Internal structure: Surface, granular brownish nucleus surrounded by
finely granular light-brown zone often almost hyalin at edge; em-
bedded, opaque with narrow, finely granular edge; bottom, opaque
or nearly opaque, brown, granular central area surrounded by light-
brown granular zone, often with narrow lighter ring at edge, the
latter often finely granular.
Edge: Surface and bottom, entire; embedded, irregularly granular or
shading off into medium.
Dextrose agar, 5 days.
Form: Surface and bottom, round; embedded, lenticular.
Size: Surface and bottom, 1 to 1.3 mm.; embedded, 0.4 mm.
Elevation: Convex.
Topography: Smooth.
Consistency: Soft.
Chromogenesis: Surface, reflected light, yellowish white; transmitted
light, translucent light brown. Embedded, reflected light, yellow-
ish white; transmitted, opaque. Bottom, reflected light, gray;
transmitted light, translucent light brown.
Internal structure: Surface, finely granular, generally with small granu-
lar nucleus. Embedded, coarsely granular; bottom, central granu-
lar area usually becoming finely granular around the edge and hay-
ing small granular nucleus.
Edge: Entire.
Beef agar, 5 days.
Form: Irregularly round.
Size: Surface and bottom, 0.7 to 1.4 mm.; embedded, 0.4 to 0.7 mm.
Elevation: Convex, often arising from depression in agar.
Topography: Smooth.
Consistency: Soft.
Odor: None.
Chromogenesis: Surface, reflected light, faint yellowish gray; transmitted
light, translucent light brown. Embedded, reflected light, faint yel-
lowish gray; transmitted light, opaque. Bottom, transmitted light,
light brown generally with small opaque nucleus and ring near edge,
Internal structure: Surface, finely granular, with round or oval dark-
brown barely translucent nucleus; sometimes growth so dense
nucleus can not be distinguished. Embedded, large central area
opaque or occasionally translucent with narrow, translucent, finely
granular edge. Bottom, granular, usually almost opaque at center.
Edge: Surface and bottom, entire or granular and blending with medium;
embedded, entire.
Potato agar, 5 days.
No growth.
266
DESCRIPTIONS OF CELLULOSE-DESTROYING BACTERIA. 39
%
Ill. Physical and biochemical features.
1. Peptone water—
Addition to peptone water.
Test for— Dex-
Lac- |Saccha-| Malt- | Glye- | Man-
trose. | tose. | rose. | ose. | erin. | nite, | Starch.
RES TOTOCTICUION® 5. pe cee ee on tty} 0 0 0 0 0 | 0 | 0
Acid production, 6 days..........-...-.. 35 - 22 05 52 0 | O .60 |
y 0 | 0 [Oe
Acid production, 12 days... ........... = esse se] s e eeee eee
Medicago hispida apiculata......-. .2s-.2p- s+ 4-0) - 22 -o---e
Medicago hispida denticulata. . 2x--Awegerin 42. bce. 2-222
Medicago hispida nigra... - ..- 0.) S62 oees oe a ee b ond ke
Medicago hispida terebellum.. . 14 42 2452 rd ed nencerscnes ncaa
Further work planned. 2. .200560css25/s50s2s-s00s 6e donee UE oT
XIII.
. Fig. 1—Pods and seeds of Medicago orbicularis.
. Fig. 1.—Pods and seeds of Medicago ciliaris.
ILLUSTRATIONS.
A heavy growth of button clover ( Medicago orbicularis) at Chico, Cal...
. Stems of button clover (Medicago orbicularis), showing appearance
characteristic-of this species._.... .<-ccrvccteresensssvenenee
. .Pods.of ten-species of Medicago. ...........«----.-DULUeStes ae
. Branch of Medicago murex, showing variation in character of pods. .
. Fig. 1—Rows of burs from single plants of Medicago, showing varia-
tion in size. Fig. 2.—Pods and seeds of Medicago radiata.......-.
. Fig. 1.—Pods and seeds of Medicago lupulina. Fig. 2.—Pods and
seeds ol: M. scutellata....cc00cesendnndesancndacuiae anne
Fig. 2.—Pods of two
subspecies of M. orbicularis.....s.00 0. 0.iso0 us ot
. Fig. 1.—Pods and seeds of Medicago rugosa. Fig. 2.—Pods and seeds
of M. tuberculata:.c) ise. J: je aol ali of 2ose Cee
. Fig. 1.—Pods and seeds of Medicago turbinata. Fig. 2.—Pods and
seeds of .M. .muricata. 2. .dccccecccecss MUU: Clee eee
. Fig. 1.—Pods and seeds of Medicago murex. Fig. 2.—Pods and seeds
Of M. rigidula... ...cccecessguccwaiscececens ot blew ie Une
Fig. 2.—Pods and seeds
. Fig. 1.—Pods and seeds of Medicago arabica and its subspecies iner-
mis. Fig. 2.—Pods and seeds of M. hispida and its subspecies
UREA So. are 3. winks 2 Bihan alts Seip Malls nici ok & alle acces ap
Pods and seeds of subspecies of Medicago hispida....:......-. Shawnie
267
Page.
10
10
20
20
24
24
28
28
30
30
32
32
34
bros:
B. P. I.—781.
NONPERENNIAL MEDICAGOS: THE AGRONOMIC
VALUE AND BOTANICAL RELATIONSHIP
OF THE SPECIES.
INTRODUCTION. .
The genus Medicago, as commonly accepted by botanists, includes ~
about 7 perennial species, with about 16 subspecies, of which alfalfa
is the best known and most important, and about 37 annual species,
with about 80 subspecies, one of which, yellow trefoil (Medicago
lupulina), has also a biennial or possibly perennial form. The dura-
tion of several—at least three—species is uncertain. There is con-
siderable difference of opinion among botanists as to the number of
annual species, mostly known as bur clovers. In 1873 Urban? recog-
nized 39 such species, with 64 subspecies, since which time 3 other
species and 17 additional subspecies have been described.
In this paper agronomic and botanical notes are given concerning
14 species and 9 subspecies which have been studied for two to five
years.
Three species are more or less cultivated or established in the
United States, namely, toothed bur clover (Medicago hispida) and
its subspecies, principally on the Pacific coast; spotted bur clover
(M. arabica), mainly in the Cotton States and in California; and
yellow trefoil, or black medic (Jf. lupulina), more or less abundant
throughout the United States. By far the greatest amount of agro-
nomic information at hand concerns these three species, and the de-
sirability of utilizing any of the other species will depend largely on
_ whether they exhibit any points of superiority.
All the annual medicagos grow under natural conditions as winter
annuals, and under cultivation they succeed best when planted in the
fall. Yellow trefoil is the only hardy species; other species can be
successfully grown only where the winters are not too cold.
In the various sections where bur clovers grow somewhat exten-
sively most of the plants are usuaily of one species or subspecies. In
California toothed bur clover (Afedicago hispida denticulata) is most
widely distributed. Jfedicago arabica, M. hispida confinis, and M.
1 Verhandlungen des Botanischen Vereins der Provinz Brandenburg, bd. 15, 1873, pp.
1-85, pls. 1-2.
267 rf
8 NONPERENNIAL MEDICAGOS,
hispida apiculata are also found in that State, but to a more limited
extent. The wide distribution of Medicago hispida denticulata in
California is partially explained by its natural adaptation, but per-
haps more by the fact that it is the most widely introduced species,
whether intentionally as pure seed for sowing for pasturage or green
manuring or unintentionally as a mixture with other seed. Medicago
hispida, M. hispida apiculata, and M. lispida confinis were in all
probability introduced into California along with W/. hispida den-
ticulata, with which they are found nearly everywhere, but in lesser
-quantity.
Spotted bur clover (Medicago arabica) is apparently of more
recent antroduction into California than W/. hispida denticulata and
is far less widely distributed in that State. On the creek pasture
lands on the Bidwell ranch at Chico, /. arabica is more often found
than M. hispida denticulata. To judge from the quantity there, it
was perhaps first introduced at this point and has been distributed
thence to various parts of the Sacramento Valley, where it is found
in small areas. According to Mrs. Katherine Brandegee, as reported
by Mr. Willis L. Jepson,! Medicago arabica is almost as common as
M. hispida denticulata in San Francisco County. Medicago arabica
is the commonest species throughout the South Atlantic and Gulf
Coast States east of the Mississippi River and succeeds exceptionally
well throughout this section. It can stand lower winter temperatures
than the toothed bur clovers (the I. hispida group), and for this
reason is better adapted to this section, in which the toothed bur
clovers more often winterkill. It is practically the only species used
for pasturage or green manuring in the Southern States. J/edicago
hispida denticulata and M. arabica succeed well in Texas, the former
species being the more generally distributed.?
Yellow trefoil (I/edicago lupulina) occurs throughout the greater
part’ of the United States, and on account of its hardiness is adapted
to sections farther north than either 1/. arabica or M. hispida and
its forms.
SOIL AND MOISTURE REQUIREMENTS.
Toothed bur clover and spotted bur clover succeed under varied
conditions as to moisture, soil, ete. In California, as well as in the
South, they grow on all types of soil from nearly pure gravel to
heavy adobe. They do better on the heavier loam soils, but will grow
in almost any soil containing sufficient moisture. They make a fair
growth even under rather arid conditions. In the dry foothill pas-
ture lands of California the toothed bur clover makes a valuable
1 Jepson, W. L. Flora of Western Middle California, 1901, p. 313,
2 Bulletin 108, Texas Agricultural Experiment Station. 1908.
267
ae
_
VALUE FOR PASTURAGE. 9
addition to the native pasturage, and in the dry-land pastures of the
valleys it is generally distributed and does well. In different parts
of Texas it is found growing along the roadsides and in waste places
where the conditions are more or less severe. It will stand a small
percentage of alkali. In California it is found on slightly alkaline
soils, but not on soils heavily charged with salts. While fairly well-
drained lands are the most desirable, spotted bur clover and toothed
bur clover produce good crops on moist lands. On California adobe
lands, which are sometimes poorly drained and lose their moisture
slowly, all three species do exceptionally well. Where there is exces-
sive moisture the crop matures later and remains green far into the
summer. While not particularly adapted to shade, both the spotted
and toothed bur clovers grow vigorously among the timber along
streams. Observation indicates that Medicago arabica is better
adapted to shady conditions than ©. hispida.
Yellow trefoil, or black medic, is somewhat notorious, from the
fact that its seed has frequently been used to adulterate alfalfa seed.
Nevertheless, the plant has agricultural merit not only as forage, but
perhaps even more as a winter cover and green-manure crop when
used in the same manner as crimson clover. The seed is usually
cheaper than that of crimson clover and the plant more hardy. At
the Arlington Experimental Farm the two plants mixed gave excel-
lent results, and yellow trefoil alone compares very favorably with
crimson clover alone.
VALUE FOR PASTURAGE.
The general characteristics of spotted bur clover and toothed bur
clover make them especially valuable for pasturage. They have
high feeding value, spread readily, and make satisfactory growth
under varied soil conditions. In the pasture lands of the South, as
well as on the Pacific coast, they have spread very rapidly after
being once introduced. The tendency of part of the seed to carry
over in the soil for several years before germination insures against
extermination by failure to develop seed in any year—from what-
ever cause, such as overpasturing or unfavorable weather. Whether
the seed germinates or carries over is apparently a matter of depth in
planting. Viable seed sown too deep will not germinate until it is
brought nearer the surface. Spotted bur clover and toothed bur
clover both contain what is known as hard seed that probably will
not germinate the first season, even if other conditions are favorable.
Such seed carries over until the second or third year.
Most bur clovers are admirably provided with means for natural
dissemination. The spiny burs of some species readily adhere to
various animals and in this way are carried long distances. Burs
65122°—Bull. 267—13——2
10 NONPERENNIAL MEDICAGOS.
without spines are distributed by other means. It is commonly ob-
served in California that orchards fertilized with manure from cor-
rals produce a heavy growth of bur clover, evidently from seed that
passed through the animals undigested and that still retained its
power of germination.
In a mixture with grasses, bur clovers are excellent pasturage and
make considerable growth of green feed during the winter and in
early spring before the grasses start. Green bur clover will often
produce bloat in cattle, and care should be taken when first turning
them into such fields. |
A heavy crop of pods and seed afford a large quantity of valuable
feed. In fact, it is the seed and pods that constitute the greater part
of the feeding value of this crop when in the dry state. Their feed-
ing value is high, and stock fed on them fatten rapidly. Pastures
containing large quantities of matured burs of the common bur clo-
vers are especially desirable for fattening sheep. The time at which
the burs are available adds importance to their feeding value, espe-
cially in California, where the dry season usually continues from
May until November. Here the valley pastures are wholly dry by
midsummer, and dry pasturage is all that is available. The bur clo-
vers, being mature, possess their greatest feeding value at such time,
but if not then fed are available later. The burs are mostly eaten
dry, but those with heavy spines are much more readily eaten after
rains have softened them.
The spiny species or subspecies of bur clovers are objectionable as
sheep pasturage on account of the burs getting into the wool. For
this reason the spineless forms are preferable. Medicago hispida
confinis is a spineless form of toothed bur clover and J/. arabica
imnermis is a new spineless form of spotted bur clover. Button clover
(M. orbicularis) is another very promising species (Pls. I and IT),
which, on account of its large spineless burs and heavy yields of seed,
is superior to the more common spotted or toothed bur clovers wher-
ever it will make an equally good growth.
SECURING AND MAINTAINING A STAND IN. PASTURES.
The seeding of pasture lands to bur clover on the Pacific coast is a
very simple process. The seed, either hulled or in the bur, is seat-
tered over the land and, without further attention, is left to itself.
Hulled seed will generally give a better stand than seed in the bur
and is to be preferred when bought. Seed in the bur is also more
likely to contain undesirable weed seeds.
In the Southern States a stand is not so readily secured, owing in
part to the fact that nodule-forming bacteria which supply nitrogen
to the plant are not present, or at least do not develop readily, except
267
of Agriculture
. Dept.
S
.
U
Bul. 267, Bureau of Plant Industry,
“WO “OOIHD LY (SIUVINOIGHO ODVOIGAI)) Y3AOID NOLLNG 4O HLMOUD AAVAH Y
ae
ky
So a ee
cy fe?
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture.
PLATE Il.
STEMS OF BUTTON CLOVER (MEDICAGO ORBICULARIS), SHOWING APPEARANCE
CHARACTERISTIC OF THIS SPECIES.
(One-half natural size. )
re
VALUE FOR HAY. 11
in the more favored and better prepared soils. It is therefore neces-
sary to supply inoculation before a good growth can be secured.
In California seeding may be done at any time during the summer,
or in the fall before the winter rains begin. In the Southern States
summer seeding is not advisable, on account of rains, and seed should
be sown in the early fall, so that it will start and continue growth
during winter. About the first of September is perhaps the best time
for seeding in most sections. When bur clover is once seeded it
persists indefinitely. Species with spiny burs, having the advantage
of this means of distribution, are perhaps more persistent than species
with smooth burs. Spiny burs are less readily eaten by animals.
Thus protected, many of them drop or are knocked from the plants,
are trampled into the ground, and thus reseed themselves. When
not trampled into the ground, the seeds of species with small burs
seem to germinate and take root much more readily than those of
species with large burs. Large smooth-podded sorts must be pas-
tured lightly or stock kept off in the spring until the seed has ma-
tured, else the immature seed is largely eaten, digested, and lost.
PALATABILITY.
As pasturage or hay, spotted bur clover and toothed bur clover are
not as readily eaten by most kinds of stock as ordinary grass and hay.
Especially in the green state, they possess a slightly disagreeable
taste, which at first makes them somewhat unpalatable. The taste
is not a serious drawback, as stock soon become accustomed to it, and
the green plants of all three species are readily eaten where other
feed is not abundant. Whether it is owing to a difference in the
palatability of the different species has not been definitely deter-
mined, but comparison indicates that in the green state most animals
apparently prefer Medicago orbicularis to M. hispida denticulata.
VALUE FOR HAY.
The bur clovers can not be considered as among the best crops for
hay. They are somewhat unpalatable, and their decumbent habit of
growth makes them difficult to handle alone. If grown with oats or
other small grain for support, they can be handled with sufficient ease
to be sometimes profitably utilized in this way. The fact that there
are better crops for hay will always limit the use of the bur clovers
for this purpose. In California, in seasons when the hay crop is
short, considerable quantities of mixed bur clover and wild-oat hay
are put on the market, but such hay is considered inferior by the
trade and sells at a lower price than grain hay. Table I gives the
chemical analyses of bur clover in comparison with alfalfa,
267
12 NONPERENNIAL MEDICAGOS.
TABLE I.—Average percentage composition of bur clover and alfalfa hay.
Kind of hay. geet Water. | Ash.
Bur clover cc. ce eeee cc ct eon 3 10.11 ie
ANgia3i22.. Mere teh: 8: 21 8.4 7.4
: Annual Report, California Agricultural Experiment Station, 1894-5, p. 147; 1896-7, p. 113.
Westgate, J.M. Alfalfa. Farmers’ Bulletin 339, U. 8S. Dept. of Agriculture, 1908, p. 28.
VALUE FOR COVER AND GREEN-MANURE CROPS.
Spotted bur clover is now used to a considerable extent in the
cotton-growing States as a winter cover crop, and its popularity for
this purpose is increasing. Over much of this area it has given better
results than crimson clover. Owing to the scarcity of seed of the
spotted bur clover, that of toothed bur clover is frequently sown;
but the results clearly show that toothed bur clover is less hardy
and in severe winters is destroyed. Toothed bur clover is commonly
used as a cover crop in California orchards, as when once well
established it volunteers from year to year. In China it is often
used as a cover crop on rice fields, and the results of preliminary tests
indicate that it will be excellent for this purpose on rice lands in
Louisiana and Texas.
Yellow trefoil has proved itself an excellent winter cover crop in
Virginia and its wide distribution leads to the belief that it has
greater merit than has been heretofore realized.
Tnoculation is of paramount importance in attaining a satisfactory
stand with any of these species.
SOURCES OF SEED.
Bur clover is little grown in the United States as a seed crop.
Spotted bur clover has been grown to some extent for seed in the
Southern States, but usually only a small acreage is so handled.
In California considerable quantities of seed of toothed bur clover
are secured with the crops of small grain, among which it grows
naturally as a weed. The seeds of bur clover ripen at nearly the
same season as the grain and are of necessity harvested with it.
The raising of grain on large areas by individual farmers, as in
California, necessitates delay in harvesting much of it, and thus
favors the development and ripening of a much larger quantity of
the bur clover than would otherwise mature. The bur clovers ripen
a little later than wheat or barley, and if these grain crops are cut
when first ripe, little bur clover seed is secured. The use of a com-
bined harvester and thrasher in harvesting and thrashing small
grain is especially favorable to the saving of bur clover seed, as in
this way a minimum number of burs is knocked from the vines.
267
eae?
TIME OF SBKEDING. 13
Bur clover seed is also obtained as a by-product from wool waste.
- Sheep running in pastures get the burs entangled in the wool and the
seeds are thus carried to the mills to be separated as a waste product.
Wool from Argentina, South America, where bur clovers are abun-
dant, contains quantities of both toothed bur clover and spotted bur
clover. This wool is shipped to the woolen mills, where the bur
clover is generally taken out as a by-product. Southern European
grazing sections are also sources of bur clover seed, which is carried
in wool as from other countries and separated at the woolen mills.
GROWING FOR SEED.
In growing a crop of bur clover for seed several difficulties are en-
countered. The prostrate growth made by the plants, the failure of
the burs to mature all at the same time, and their tendency to drop
very easily from the stem as soon as ripe make the harvesting of seed
dificult. To grow bur clover as a seed crop on a large scale is most
practical in sections having a continuous dry summer. Rains in
summer are apt to cause the seeds to germinate in the burs, making
them more difficult to handle; but where such rains occur it is both
practicable and advisable to raise seed in small quantity for one’s
own use.
The fact that the greater part of the commercial bur clover seed
is Medicago hispida and its forms, with little WM. arabica, makes it
_ almost necessary for the farmer in the Southern States to grow his
own seed at the present time. As has been stated, I/. arabica, accord-
ing to our present knowledge, is the best species for the South.
Before seeding, the land should be put in as good condition as pos-
sible by plowing and harrowing, and if the seed is to be harvested by
any method such as sweeping, the field should be run over with a
float or roller to leave a smooth surface in order to facilitate the
harvest. If a drill is used to sow the seed the ground should be
especially well firmed.
TIME OF SEEDING.
In sections having a mild winter climate, bur clover should be sown’
in the fall. In California, where dry weather prevails throughout the
summer, the seed may be sown at any time before the fall rains begin.
Where summer rains occur, as in the Southern States, the planting
should be delayed until the first of September. In dry sections, where
it is desirable to start the seed in the fall with irrigation, the plant-
ing should be done about the first of October. The object is to sow
the seed so late that a subsequent irrigation will not be necessary.
Summer seeding in the South is not advised, as the young plants
starting at that time are liable to suffer from drought, and where a
267
14 NONPERENNIAL MEDICAGOS.
heavy growth is made the plants tend to mature and die rather than
continue growth through the winter. When seed in the bur is used,
earlier planting may be practiced than when hulled seed is used.
Germination will be delayed on account of the protection afforded
by the burs, and the result is the same as a later planting of hulled
seed. .
METHOD AND RATE OF SEEDING,
The clean seed may be sown broadcast or by using an ordinary
grain drill with press-wheel attachment. Special care should be
taken to cover the seed thinly. The drill should be used only on well-
firmed soil, as otherwise the seed will be planted too deep. The
press-wheel attachment is necessary for the best success when a drill
is used. In general, broadcast seeding will perliaps be found the most
satisfactory and is the only method that can be employed when the
seed is sown in the burs. A light harrowing is all that is necessary
to cover hulled seed sown broadcast and will usually cover seed sown
in the bur. When the land is left with light furrow markings, such
as are made by a large-toothed harrow, seed not covered by the har-
row at the time of seeding will fall into these small furrow depres-
sions and be covered by the washing of subsequent rains. Good
stands have been secured by this method without covering the seed at
all at the time of sowing, and it probably will be found satisfactory
in sections where a continuous rainy season occurs.
INOCULATION.
In the Southern States inoculation of the land is necessary to grow
clover for the first time. In California the soil apparently is in
most places already inoculated. The best method of inoculation is
perhaps to mix a small amount of soil from an old bur-clover field
with the seed, whether hulled or in the bur. The quantity of soil
used need be only a mere dusting. Sowing seed in the bur seems also
to insure inoculation, and for this reason it is commonly practiced
in the Gulf Coast States. Open and loamy soils are most easily
inoculated, and it is recommended that to establish bur clovers on a
place an old garden patch or other well-prepared and manured piece
of land be selected.
HARVESTING AND THRASHING,
The limited work that has been done with bur clovers has not en-
tirely demonstrated the best method of harvesting the seed but has
at least indicated the difficulties to be overcome and has suggested
improvements on methods used. Several processes have been tried.
The combined harvester and thrasher has been used in an attempt to
267
i
be
i
HARVESTING AND THRASHING, 15
eut and thrash the crop direct from the field at one operation, as
small grains are harvested in the West. The seed of bur clover ripens
continuously through a long period; hence, a large quantity of un-
ripe burs are harvested even if the crop is cut when the yield is at
its maximum. The green burs and accompanying green portions of
the stems, which are gathered with the ripe burs and seed, contain
much moisture and without special drying are likely to heat before
they can be taken to a huller and the seed separated.
If the bur clover is sown with a grain crop which is allowed to
ripen thoroughly, the difficulty just mentioned is largely overcome,
but the yield of seed by this method is small, because the burs
drop from the plant so easily as soon as ripe. It is also necessary
to let the grain crop become overripe in order to allow the bur clover,
which matures a little later, to develop its maximum yield of seed.
The use of a common self-rake reaper has been suggested but not yet
tried. The idea is to cut the crop when a maximum amount of seed
is ripe and then to handle it as the seed crop of red clover or alfalfa
is handled. The use of the reaper should reduce the loss of pods
to the minimum possible with ordinary farm machinery, but whether
the method is practicable remains to be demonstrated. The readi-
ness with which the burs drop from the plants will make this method
difficult at best, but by operating at a favorable time, as on a cloudy
day or early in the morning, the loss of burs will be reduced to a
minimum.
The idea of air suction has been tried in an attempt to overcome
the difficulty occasioned by the burs dropping from the vines. This
process has been tried by Mr. R. W. Jessup, of Oakland, Cal., who
reports it only a partial success. A power suction machine was used
and 20 acres of burs harvested. The vines were allowed to become
thoroughly dry and were then cut with an ordinary mower and
raked into windrows. The ground was thus left comparatively clean
and in shape for the suction machine to operate. In the process of
mowing and raking, all the burs were knocked from the vines, so that
2 maximum yield was obtained. By this method a quantity of other
substances, such as sticks and small stones, were gathered with the
burs and were very objectionable on account of the damage to the
cylinder in the process of hulling. To overcome this objection a
special device for cleaning foreign substances from the burs as they
were harvested was used in connection with the suction machine in
1911. This device made the machine more satisfactory, but the
method at best is somewhat expensive. With a heavy yield of seed
the expense is reduced.
A method of harvesting first employed in the West and South is to
allow the seed to ripen thoroughly and then to cut the vines with an
267 |
,
16 NONPERENNIAL MEDICAGOS.
ordinary mowing machine and rake them into windrows. The burs
are then swept together with large barn brooms and hauled from the
field. The burs gathered in this manner are mixed with more or less
gravel and other foreign substances, which must be removed before
the seed can be satisfactorily hulled or used in the bur. This separa-
tion is accomplished by the use of handbarrow screens and an ordi-
nary fanning mill regulated to blow the burs over; or, if running
water is handy, a quicker and more satisfactory metab is to throw
the burs into the water. All heavy substances sink, and the burs and
lighter substances are dipped from the stream. To facilitate this
method of separation the channel of the stream should be narrowed
in the shape of an open V, which generally aids in collecting the
cleaned burs. To dip the burs from the water a large handbarrow,
with a bottom made of wire netting, has been found very satisfac-
tory. The burs are spread on canvas to dry, after which they are
ready for the huller.
All bur clovers, whether with large hard burs, like Medicago tur-
binata, or small soft burs, like /. hispida denticulata, or large soft
burs, like M. orbicularis, are successfully hulled with an ordinary
clover huller.
YIELD OF SEED.
Few data as to the actual seed yield of the various species of bur
clovers are available. Table II gives the results of tests made at
Chico, Cal.
During the winter of 1907-8 seasonal conditions were rather un-
favorable to the production of heavy yields of seed, and the figures
given (Table II) are umdoubtedly somewhat lower than may be ex-
pected in a more favorable year. In the test referred to, the seed
was sown in the fall before the winter rains began, and the crop was
allowed to develop under natural seasonal conditions, without irriga-
tion.
During the winter of 1908-9 Medicago orbicularis, M. hispida
nigra, and M. hispida confins were again grown for seed in one-
twentieth acre plats. In this test the seed was sown early in October,
with irrigation at time of seeding only. A good winter growth was
thus insured. The yield of seed in the irrigated plats (Table IT)
was considerably greater than in the nonirrigated plats, which
amounted to little.
During the winter of 1909-10 Medicago orbicularis, M. scutellata,
M. hispida confinis, M. hispida nigra, M. turbinata, and M. hispida
terebellum were again grown in one-twentieth acre plats. They were
sown in October, 1909. The plats were irrigated before seeding only.
The seed yields are not entirely comparable, on account of variation
267
RELATION OF WEIGHT OF BURS TO THEIR VOLUME. 17
in the stands germinated. The plat of U/. hispida terebellum had a
very poor stand, and those of UY. turbinata, M. hispida nigra, and
M. orbicularis microcarpa (No. 7738) were thin. M. orbicularis
(No. 10725), MW. hispida confinis, and M. scutellata had good stands.
The large seed yields of Medicago orbicularis, which is a very
promising species, have been very consistent through the four years.
Hulled seed of bur clover weighs about the same as alfalfa seed—60
pounds to the bushel.
TABLE II.—Yields of seed per acre at Chico, Cal.
Yield of hulled seed per acre.
8. P. I. No.! Species tested.
i 1903 1909 1910 1914
Pounds. | Pounds. | Pounds. | Pounds.
Maes ones Medicago orbicularis ................ we Sateen 860 790 1,160 947
i... eas Medicago orbicularis microcarpa.. ...-- ees S204 SOR Vadis - RE Nance aiutiokin 2%
0 ee Medicago hispida terebellum ................- Ce Se en ees Oe eS ee
yy Medicago scutellata..........- a A ee ZAO" fe eer (SU (Bas dete anes
Ve (re Medicago turbinata ........... ety SS gee ae 220 s\ oes 180) tS oes-cemes
Met iae..-.-.-.- Medicago hispida nigra .............-- ec hey 470 440 a Ce
See Medicago hispida confinis ...............-.-.. 320 375 Ce ee ‘
Medicago hispida denticulata ................ BO) he. = a = Shad ae icant hae et =
1 Seed and Plant Introduction number.
RELATION OF WEIGHT OF SEED TO VOLUME AND WEIGHT
OF BURS.
The weight of seed in a given volume of burs varies considerably,
mainly owing to differences in bulkiness of the burs in the several
species. These variations are due not entirely but largely to differ-
ences in length of spines. Species with long spines have less seed in
a given volume than spineless forms, especially when closely related
types are compared.
The weight of seed in a given weight of burs also varies somewhat
in the different species, mainly owing to differences in the texture of
ihe burs. The harder types of burs have the smaller percentages of
seed. Table III shows that the weight of seed in a bushel of burs
in the different species varies from 1.75 pounds in J/edicago arabica
to 4.66 pounds in M. turbinata; and that the weight of seed in 100
pounds of burs varies from 20.89 pounds in J/. turbinata to 33.78
pounds in W/. hispida denticulata.
RELATION OF WEIGHT OF BURS TO THEIR VOLUME.
The great differences in the spines and in the texture of the burs
make decided differences in the weight of burs in a given volume in
the several species. A given volume of a species having very short
or no spines or of those with hard burs is much heavier than of
species having long spines and soft burs. Considerable variation in
65122°—Bull. 267—13-——8
18 ‘NONPERENNIAL MEDICAGOS.
weight is caused by packing the burs, especially of the spiny species,
and for this reason the weights can be only approximated. Table III
shows that the weight of a bushel of burs in different species varies
from 6 pounds in Medicago arabica to 22 pounds in M. turbinata.
TABLE III.—Relation of weight of burs and seed to volume.
Weight Weight of seed.
of 1
8. P.I. No. Species tested. aed lad In 100
Bore: bushel | pounds
of burs. | of burs.
Pounds. | Pounds. | Pounds.
14 3. 83
P11 eee ee ee Medicavoiscntellatay. - . Sosseee. se eases 2 Le eee. F 27.28
07a Se Medicago'orbicularisi.;- 5) $2.2 22 cue. ose ceet eee ee eee 8 2. 66 33.5
ONIGl«. 1. Le Medicago ‘tur binatats2 tee: Sif) eee AB hls 22. 33 4.66 |" 20. 89
7.3 UF § eae Sean: Medicago’ hispida nigra 2.0.2.2... cand eons oes seen 8 2 25
MAD ook Medicavo eiliaris:.}.20&. 2 8 eee ee eee 8. 66 2.16 25
RGR7O ses. 5 oc. Medicago hispida terebellum ........-..-.2<..-2..2--0--2- 11.99 3.18 26. 39
Mediegps deutiowiate .3).. 24. sca. dtc secs dete ees | 6.16 2. 08 33.78
1.75 29.16
Medicago'arabica.:-- ...23.0.. 2. ee ee ae eos 6
RESEMBLANCE TO ALFALFA SEED.
The seeds of a number of species of bur clover resemble alfalfa
seed very closely. The most common are the Medicago hispida group,
M. lupulina, and M. arabica, the seeds of which are of lighter yellow
color, lacking the rich, greenish yellow shade of alfalfa; and all are
uniformly larger except J. lupulina, which is somewhat smaller and
is the only species in which the difference in size is readily noticeable.
Medicago arabica is further distinguished by having a small, well-
developed projection at the end of the hilum.
VITALITY OF SEED.
As mentioned elsewhere, bur clover seed retains its vitality for a
very long time. Seed three years old will generally show delayed
germination, but it is only after several years that the percentage of
germination is noticeably decreased. (Table IV.) Not only does
seed retain its power of germination when kept under dry conditions,
as in a seed room, but it will carry over in the soil for a number of
years in the same way.
TaBLe 1V.—Germination tests of Medicago arabica.*
“ : ioe ae yf Fy ~~
ge o uration tion 0 ion 0
8. Pe. No. seed. of test. unclipped Hard seed. Good seed. clipped
seed. seed.
Years, Days. Percent. | Percent. | Percent. | Per cent.
169180... ccpnenctasinandcasbans 6 12 20 60 80 77
ZIBGO. Joechatdduecedeuewreeuse 4 12 30 65 95 90
WI0G1 . . Scigeccdatentsneyunehle 3 7 26 53 79 77
Rie sekeacecusetebecerackuas 2 7 19 76 95 94
' Results obtained by the Seed Laboratory, Bureau of Plant Industry.
267
-' ( <3. eget ee,
DIFFERENCES IN THE BURS OF DIFFERENT SPECIES. 19
It has been observed in California, in orchards kept free from all
growth during the spring months and no seed allowed to develop,
that a good stand of the toothed bur clover occurs each year for four
or five years after the last crop of seed was allowed to mature in the
orchard. ;
Plantings of a number of species of bur clover were made in pots so
that this point could be more definitely observed and showed that it
was a common occurrence for seed to carry until the second year be-
fore germinating, even when the depth of planting and other con-
ditions were favorable for growth.
INSECT ENEMIES.
The clover-seed chalcis fly (Bruchophagus funebris How.) which
attacks red clover’ and alfalfa? is also common in bur clover. The
small flylike insect lays its eggs in the ovules; the larve develop in
the seed and reach maturity by the time the seed is ripe. The amount
of seed thus destroyed at Chico, Cal., is considerable, especially in
that maturing late. Of the early-maturing seed perhaps 10 per cent
is destroyed, while the loss of late seed may be as high as 75 per cent.
All species tested are subject to its attacks, some more severely than
others. No practical way of controlling this pest seems to be known.
DIFFERENCES IN THE BURS OF DIFFERENT SPECIES.
The pods, or burs, of the different species of annual medicagos
differ very much in size, form, and with regard to the spines (PI.
III). They also vary widely in weight and texture.
In such species as Medicago orbicularis and M. scutellata the burs
are very large and spineless, being decidedly flattened in /. orbdicu-
laris and nearly spherical in UM. scutellata. The pods of both are
soft and somewhat papery. In Medicago ciliaris and M. echinus the
burs are very large and have heavy spines. The spines are erect in
the former and decidedly appressed in the latter species. The texture
of the bur tends to be hard in M/. ciliaris and a little less so in WV.
echinus. ‘The general form in both species is oval. In MU. turbinata
the bur is large, oval, very hard (in the most common type), and has
a few short tuberclelike spines. Medicago rigidula and M. murex
are somewhat similar to M/. turbinata, but they are smaller and com-
monly have longer spines. Of species with smaller burs, some, as J.
hispida denticulata and M. arabica, have spines, and some, as M..
hispida confinis, are without spines. All variations between these
types are found, and there are many other forms which mark
botanical characteristics peculiar to definite species.
1 Circular 69, Bureau of Entomology, U. S. Dept. of Agriculture, 1906, p. 7.
* Farmers’ Bulletin 339, U. S. Dept. of Agriculture, 1908, p. 41.
267
20 NONPERENNIAL MEDICAGOS.
DESIRABILITY OF A BUR WITHOUT SPINES.
For various reasons a bur without spines is more desirable than
one with spines. Spineless burs do not catch in the wool of sheep,
an objectionable feature of ordinary bur clovers. On the other hand,
they may be objected to on account of being deprived of this means
of distribution, as the smooth burs will not hang to stock to be car-
ried about and are a little harder to maintain in pasture, especially
the larger podded varieties. Furthermore, the spineless burs are
more readily eaten by stock. As already explained (p. 10), the weight
of evidence favors the smooth bur.
STUDIES RELATING TO VARIATION IN THE BURS OF DIFFERENT
SPECIES.
Since the fall of 1908 a large collection of bur clovers has been
used in a study to determine to what extent the burs of the various
species and subspecies vary from their normal type. This collection
comprises 202 selections, and includes 20 species and subspecies.
It may be well to mention here a few difficulties encountered in the
work. Like the seeds of many other legumes, fresh seeds of bur clover
do not germinate readily. A common experience has been that the
entire lot of seed of a selection failed to germinate. Some difficulty
has been found in so protecting the plantings as to be perfectly sure
that mice had not carried seed from one selection to another. To
get soil absolutely free from bur clover seed is somewhat difficult
where bur clover is naturally abundant, and it has necessitated extra
care. The first year all the soil used was sifted through screens
sufficiently fine to exclude any bur clover seed that it might contain.
In the second and subsequent years the soil used was taken from 4
feet below the surface of the ground, at which depth it was found
to be free from all germinable seed. The latter method is satisfactory
at, Chico, Cal., where the work has been carried on. Plants grown in
soil from a depth of 4 feet have been found to make a growth quite
as good as plants grown in soil taken at the surface.
In order that the comparison between the original burs selected
and the burs of their progeny might be as accurate as possible, burs
of each selection from which seed was taken for planting (or burs
as nearly like the type as could be found) were saved for future com-
parison. In the descriptive records of the plants the following points
were observed: (1) Pubescence, (2) leaf markings, (3) size and
color of leaflets, (4) size and number of flowers, (5) size, color, and
shape of stems, and (6) general notes. Besides descriptive notes,
typical burs produced in each season were saved, together with her-
barium specimens of most of them, so that comparison of any varia-
tion in the progeny from the original selections could be noted.
267
rer
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. aD PLATE Ill.
PODS OF TEN SPECIES OF MEDICAGO.
Top row, JW. arabica and M. hispida denticulata; second row, M. hispida conjinis and
M. hispida terebellum; third row, M. muricataand M. hispida nigra; fourth row, M.
ciliaris and M. echinus; bottom row, M. scutellata and M. orbicularis. (Natural
size.)
Bul. 267, Bureau of Plant Industry, U. S. Cept. of Agriculture. PLATE IV.
BRANCH OF MEDICAGO MUREX (NO. 0147), SHOWING VARIATION IN CHARACTER OF Pops.
— -_ a
VARIATION IN THE BURS OF DIFFERENT SPECIES. 21
The plants have been grown each year in 77-inch pots. The piant-
ings have been made each year in the fall, and the first year (1908-9)
the plants were carried through the winter in a cool greenhouse. The
second year (1909-10) the plants were carried through the winter
in an ordinary lath house, and the third year (1910-11) in a glass-
covered lath house. Every year in the spring the pots were plunged
in soil to their entire depth and allowed to remain in the lath house
until the seed was mature.
VARIATION IN MEDICAGO MUREX.
From plantings made in the fall of 1908 a very marked variation
occurred in two selections (F. C. I. No. 0147 and S. P. I.t No. 16875)
which had been received under the names Medicago polycarpa and
M. murex, respectively. A single plant of each produced burs vary-
ing not only in shape, but ranging from spineless to forms having
many medium-sized stout spines (Pl. IV). The variation with re-
gard to spines was as great as could be. F.C. I. No. 0147 and S. P. L.
No. 16875, which had been received under different specific names,
proved to be identical, both being J/. murex. In their first season
and in subsequent years the plants of these two numbers were noted
as being identical in general growth, size, shape, color, and markings
of stems, leaves, and flowers.
In the fall of 1909 plantings were again made as in 1908, but this
time seed from the progeny of the 1908 plantings was used. Burs
of F. C. I. No. 0147 and S. P. I. No. 16875, both smooth and spiny
and also including intermediate forms, were planted. Each series
represented all types of burs taken from a single plant. The result-
ing plants from the varying types of burs were all alike, and they
were like the plants of the year before. The following spring (1910)
these plants fruited, and all the early burs were spiny and essentially
like the spiny forms produced in the spring of 1909. To see whether
adverse conditions would produce such a variation in the burs as
occurred in 1909, pots containing the plants were lifted out of the
ground in which they had been plunged, to cause them to dry out
more readily. Abnormal or spineless burs began to form at once
on the plants thus lifted. A week later a hot spell of a few days’
duration dried all the plants severely, and spineless burs began to de-
velép also on the plants not lifted. From that time on, hot weather
prevailed and varying burs continued to form. (See Pl. X, fig. 1.)
In the fall of 1910 plantings were made again as in previous years.
In the spring of 1911 pots were lifted as before, with the same gen-
eral results. The burs without spines produced in 1911 were not as
well developed as in 1909, but showed the same general variations.
FR, €. Fis aa Abbreviation toe’ Wotece Crop Piyedtign tone « S. =, re Foreign Seed
and Plant Introduction.
267
“29 NONPERENNIAL MEDICAGOS.
VARIATION IN MEDICAGO CILIARIS,
In the spring of 1910, at the time of lifting the pots of Medicago
murex, pots of WM. ciliaris were also lifted, to note the effect on burs
of this species. Up to this time all the burs developed had been nor-
mal. In two or three days after lifting the pots the burs began to
show elongation. Burs subsequently produced were decidedly elon-
gated, and the spines were much shorter than normal. In the pots
not lifted the plants continued to produce normal burs until severely
checked by a hot spell some time later, after which abnormal burs
were produced on all the plants. (See Pl. XI, fig. 1.) In the spring
of 1911, pots of this species were again lifted as in the previous year,
with the same general results.
—
VARIATION IN MEDICAGO MURICATA,
The pots of Medicago muricata were not lifted as were those of
M. murex and M. ciliaris, but in the seasons of 1909, 1910, and 1911 it
was noted that the burs formed late in the season varied greatly with
regard to spines. In some instances the burs were nearly smooth
and varied from this type to nearly normal burs. The burs formed
earlier in the season were always normal and spiny. Plantings of
this species had been made in the open field in the fall of 1910 and
were observed to determine the extent of variation under field condi-
tions. The results were practically the same as with the pot-grown
plants. The burs which developed early were normal and with
spines, while those formed late in the season varied from nearly
smooth to nearly normal. (See Pl. IX, fig. 2.)
VARIATION IN OTHER SPECIES.
Besides Medicago murex, M. ciliaris, and M. muricata.the following
species were planted in the fall of 1908 and the two succeeding years:
M. scutellata, M. orbicularis, M. orbicularis microcarpa, M. orbicu-
laris marginata, M. hispida, M. hispida nigra, M. hispida confinis,
M. hispida apiculata, M. hispida denticulata, M. hispida terebellum,
M. echinus, M. turbinata, M. tuberculata, M. rugosa, M. lupulina,
M. radiata, M. arabica, and M. intertexta.
Within each of these species, types and variants were selected
from a general lot of burs and the seed planted, to note variations in
the progeny. In none of these species was there the marked variation
of burs on single plants noted in Medicago murew and M. muricata,
but in every species the burs on individual plants varied somewhat
in size and the spines varied more or less in length. In M. hispida
and its subspecies the variation was so great that the smallest burs
produced in the different subspecies were smaller than the typical
267 |
_* ey
VARIATION IN THE BURS OF DIFFERENT SPECIES. 23
burs of the nearest related subspecies having a smaller bur. (PI. V,
fig. 1.) In all cases the number of burs of the progeny that varied
from the type of the species or subspecies was much less than the
number of those that were typical.
It has been noted that burs of the same species vary in color, rang-
ing from very dark or almost black to straw color. General obser-
vations had indicated that the dark color might be due to moisture
in contact with the burs before and after ripening. That the dark
colors were due to such contact was demonstrated very clearly on a
large scale in the spring of 1911 at Chico, Cal. Until the burs were
ripening the weather had remained clear and dry, and the burs de-
veloped were all light colored. Then a light rain fell during one
night, and the next day all the burs that were fully developed and
- ripening (which included the greater part of the crop) turned black.
No other rain fell, and all the burs that matured later were light
colored and remained so. _
The following species growing in the open at Chico at the time
of the rain referred to were noted as showing a similar. change in
color: Medicago orbicularis, M. orbicularis marginata, M. ciliaris, M.
scutellata, M. turbinata, M. muricata, M. tuberculata, M. rigidula, M.
hispida, M. hispida confinis, M. hispida apiculata, M. hispida tere-
bellum, M. hispida nigra, and M. arabica. The discoloration in I/.
arabica was not as marked as in the others.
To test this phenomenon artificially, burs of Medicago orbicularis
that had developed without becoming wet or discolored were damp-
ened and left over night; the next morning all had become dark,
with the exception of a few burs that were mature and dry. It
would seem from these observations that the dark-colored burs in
all species are probably those that have come in contact with moisture
during the period of their ripening and before they are fully mature.
The color of the burs can not be used as a character on which to base
botanical subspecies.
VARIETAL STRAINS
To determine whether varietal strains exist in different species or
subspecies, a number of selections were made representing types with
regard to length of spines and size and form of burs. Two types of
burs of Medicago scutellata were selected and planted. In one type
the bur was so coiled as to have a definite truncate end, while in the
other the bur was more nearly conical. AJ] the progeny of the trun-
cated type produced truncated burs, and by far the greater part of the
progeny of the conical burs produced conical burs and only a few.
tended to be truncated, as shown by the bulk of burs harvested.
267
94 NONPERENNIAL MEDICAGOS.,
A large number of burs of Medicago orbicularis and M. orbicularis
microcarpa of various types were selected—double convex, convex on
one side only, and burs open (or loosely coiled, as in /. orbicularis
marginata). The seed from the selected burs was grown in the field
in plats. Most of the burs from the progeny plants were like the type
planted, as shown by the bulk of burs harvested.
In the Medicago hispida group, burs representing the different sizes
and lengths of spines were planted. In a number of cases they re-
produced true to the type selected. M. hispida confinis produced two
types, the bur of one having 23 to 3 turns and the other 4 to 44 turns.
Another type approaching that of W/. hispida confinis, but with spines
sufficiently developed to throw it out of that group, reproduced true to
type. In the other groups of I/. hispida the types were not so defi-
nitely marked, but a few showed variations. The other species
showed no definite varietal strains, but the work was not extended
enough to say that they do not exist. Thus far the work indicates
that there exist definite varietal strains within at least several of the
species and subspecies, and that these may be grown as pure strains
by selection. On account of the variations that may occur within
the different species, a type or variety can not be defined by the ap-
pearance of the burs in a bulk lot of seed. As already explained,
an individual plant may produce burs that are as different as the burs
of two subspecies, but when grown under normal conditions most of
the burs on individual plants will be true to type.
~~
POLLINATION IN THE VARIOUS SPECIES.
The flowers of the various species of bur clovers are similar in
form but differ somewhat in size, in the number borne in a cluster,
and in the details of the explosive mechanism. Seven of the species
studied have a tripping mechanism similar to alfalfa, so that after
tripping the stigma is exposed—-Medicago scutellata, M. rugosa, M.
turbinata, M. muricata, M. rigidula, M. ciliaris, and M. echinus. In
M. echinus the flowers are in clusters of six. The other species have
the flowers in clusters of two. Medicago turbinata often appears to
have single flowers in a place, because one of each pair usually dies in
the bud or withers shortly after it opens and fails to develop a pod.
All the bur clovers studied except Medicago echinus seem to be
readily self-fertile. The various species have been grown in a green-
house where little, if any, cross-pollination was probable, and with the
exception of Jf. echinus all set pods freely. Alfalfa plants growing
in the greenhouse beside the bur clovers set no seed except when
artificially tripped. A number of flowers of J/. echinus were tripped
by means of a toothpick to determine the effect on seed setting. These
267
Bul. 267, Bureau of Plant Industry, U. S, Dept. of Agriculture.
Fig. 1.—ROWS OF BURS FROM SINGLE PLANTS OF MEDICAGO, SHOWING VARIATION IN
SIZE.
Upper row, JV. hispida nigra; middle and lower rows, M. hispida denticulata. (Enlarged 23
diameters. )
Fig. 2.—PODS AND SEEDS OF MEDICAGO RADIATA, SHOWING SEED AND VENATION OF PobDs.
(Enlarged 2 diameters. )
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE VI.
Fia. 1.—PODS AND SEEDS OF MEDICAGO LUPULINA, SHOWING VENATION OF PODS AND
PROMINENCE ON THE SEED AT THE TIP OF THE RADICLE.
(Enlarged 9 diameters. )
Fic. 2.—PoODS AND SEEDS OF MEDICAGO SCUTELLATA, SHOWING VENATION AND WINDINGS
OF PODS AND CHARACTERISTICALLY NOTCHED SEEDS.
(Enlarged 2 diameters. )
GENERAL CLASSIFICATION OF THE SPECIES. 25
flowers, as well as the alfalfa flowers, set a number of pods, but the
exact percentage was not determined. Other flowers were tripped
and cross-fertilized with pollen from another plant of YU. echinus,
with the apparent result that more pods were set from these flowers
than from those tripped but not cross-pollinated.
GENERAL CLASSIFICATION OF THE SPECIES.
The nonperennial species of Medicago here considered may for
convenience be divided into six groups. The most of the species in
each group are so nearly alike in flower and leaf characters that they
are distinguished with certainty only when the pods have matured. ©
The first group contains a single species, Medicago radiata, the
leaflets of which are mostly rather small and the stems somewhat
woody, procumbent to erect. The pods are large, flattened, kidney
shaped, and have a row of short, simple or sometimes forked spines
along the back. The seeds have the surface somewhat convoluted
and the radicle as long as the seed.
The second group contains one species, Medicago lupulina, having
stems slightly procumbent or suberect in habit, with rather small,
strigose-veined leaflets and small kidney-shaped pods.
The third group contains three species, Medicago orbicularis, M.
scutellata, and M. rugosa. ‘The stems in this group, as in the follow-
ing three groups, are more procumbent and, unless the stand is thick,
have a tendency to become trailing. The pods are rather large, disk
shaped, consist of several thin spiral windings, are of a papery
texture, and are without spines.
The fourth group contains four species, Medicago rigidula, M.
turbinata, M. tuberculata, and M. murex. The pods of these species
are harder in texture, more closely wound, of an oval form, and vary
from smooth to tubercular and spiny.
The fifth group contains two species, Medicago ciliaris and M.
echinus, which have rather large, closely wound, oval pods, with
windings the edges of which are thickly covered with interlocking
spines. |
The sixth group contains two species, Medicago arabica and M.
hispida, which have smaller, somewhat short cylindrical pods, with
windings the edges of which are usually covered with more or less
erect spines. Forms of both species are found without spines.
In the last four groups the flower and leaf characters are so nearly
alike that the species can be distinguished with certainty only by
the mature pods.
The seeds vary from 2 to 6 mm. (one-sixteenth to one-fourth inch)
in length, and in the species here considered are yellow or greenish
yellow in all but three species, Medicago ciliaris and UM. echinus hav-
267
296 NONPERENNIAL MEDICAGOS,
ing black seeds and MW. orbicularis having yellowish brown seeds. In
all but two species here considered the seeds are kidney shaped and
smooth, the exceptions being I/. radiata, in which they are oval in
outline with a convoluted surface, and M. orbicularis, in which they
are obovoid and the surface papillose.
Many of the species upon which these studies are based were origi-
nally obtained in Algeria by Mr. C. S. Scofield in 1901. Medicago
orbicularis, S. P. I. No. 10725, is also from Algeria, secured in 1902
by Mr. T. H. Kearney. The others are mainly from botanical gar-
dens, especially that at Madrid, Spain, and a few are from miscella-
neous sources.
DISTRIBUTION AND DESCRIPTION OF SPECIES.
The genus Medicago is at present widely distributed over southern
Europe, western Asia, northern Africa, and the adjacent islands.
Its northern limit seems to be southern Scotland, Sweden, and Siberia.
A few species have become naturalized in recent years in Abyssinia,
South Africa, and Chile. In the United States about 60 species and
subspecies have been introduced. for experimental purposes since
1898 through the Office of Seed and Plant Introduction. Some of
these importations may become naturalized. Prior to 1898 five species
had become well established along the eastern, western, and southern
coasts of the United States. These species and two others more
recently introduced, are gradually working their way inland, but
their progress is slow. At present but two species are established in
the Central States north of Texas.
MEDICAGO RADIATA L,
(Pl. V, fig. 2.)
Stems decumbent, pubescent, 10 to 30 cm. (4 to 12 inches) long; leaflets
obovate to cuneate, downy to villous on both sides, 2 to 6 mm. (one-sixteenth
to three-sixteenths inch) wide, 3 to 10 mm. (one-eighth to three-eighths inch)
long, rounded, and toothed at the apex, the base entire; leafstalks equaling or
twice the length of the leaves, pubescent, the stalk of the terminal leaflet five
times longer than the lateral; stipules awl shaped, pubescent, entire, 2 to 4 mm.
(one-sixteenth to one-eighth inch) long; flowers usually axillary, in clusters of
two, 24 to 3mm. (three thirty-seconds to one-eighth inch) long, the stigma not
exposed when tripped, the peduncles and calyx pubescent ; pods papery, brownish
when ripe, sickle shaped to circular, 15 to 25 mm. (one-half to 1 inch) long, 7 to
10 mm. (one-fourth to three-eighths inch) wide, glabrous, netted veined, with
a row of simple, sometimes forked spines about 1 mm. (one-sixteenth inch)
in length along the outer side, and an irregularly toothed, membranous margin
along the inner side, 5 to 6 seeded; seeds oval, flattened, light to yellowish
brown, 2 to 24 mm. (one-sixteenth to three thirty-seconds inch) long, surface
convoluted, the radicle as long as the seed. i
Distribution: Spain to Persia.
267 :
OT ee
DISTRIBUTION AND DESCRIPTION OF SPECIES. 27
This species was received from Madrid, Spain, and Karlsruhe, Germany,
under S. P. I. Nos. 9746 and 16266, respectively. It has been tested only at
Chico, Cal. It makes little growth compared with toothed or spotted bur clover,
and yields little seed.
MEDICAGO LUPULINA L. (YELLOW TREFOIL).
(Pl. Vi) ag)1.)
Stems four angled, pubescent to nearly glabrous, 10 to 80 cm. (4 to 32 inches)
long, decumbent; leafstalks, 2 to 10 cm. (three-fourths to 4 inches) long, pubes-
cent, the leaflets oval to broadly obovate or even obcordate, sometimes wedge
shaped at the base, pubescent on both sides, 3 to 12 mm. (one-eighth to one-half
inch) wide, 6 to 20 mm. (one-fourth to three-fourths inch) long, the base entire.
stalk of the terminal leaflet 3 to 5 times longer than the lateral; stipules rather
large, broad, and few toothed at the base; flowers very small, 13 to 2 mm. (one-
sixteenth inch) long, in rather close, oval, or oblong heads of 10 to 40 flowers,
the stigma not exposed when tripped; pod kidney shaped, about 2 mm. (one-
sixteenth inch) in diameter, netted veined, minutely pubescent, blackish when
mature, one seeded; seed 13 to 2 mm. (about one-sixteenth inch) long, yellow
or greenish yellow.
Distribution: Spain to Scotland, east to Persia, and probably widely intro-
duced into every civilized country.
This species is a semierect, leafy plant, usually biennial, but with annual and
perennial forms. It occurs spontaneously both in the Pacific Coast States and
in the Eastern States. It is hardy at least as far north as central New York
and, in addition to its use as pasturage, is of promise as a green-manure crop.
It is being tested in comparison with crimson clover in the Atlantic States, and
while its actual yield of forage and green manure is not equal to crimson
clover, the occasional high price of the seed of crimson clover makes this species
potentially important. At the Arlington Experimental Farm, Va., it made a
growth varying from 12 to 26 inches in height, depending on the character of the
soil. It will grow on stiff clay soils somewhat too poor for the successful growth
of alfalfa or red clover. It has been recommended as a constituent of lawn mix-
tures, since it remains green during rather severe drought. At Chico, Cal.,
it has been grown under F. C. I. No. 0268, seed received from the Botanical
Gardens af Madrid, Spain, and also under S. P. I. No. 4340, from Naples, Italy.
In hot weather it made considerably better growth than the other bur clovers
under test. It is used to some extent in European pastures and is ordinarily
regarded as being inferior to clover and alfalfa. Its use in the past to adulter-
ate alfalfa and clover seed has caused it to be classed as a weed, but it is not
troublesome in this respect, since it is readily eradicated by ordinary tillage.
Its presence in uncultivated lands is not objectionable, but usually advantage-
ous. Its short life makes it compare unfavorably with alfalfa, and its small
size makes it less valuable than such plants as crimson clover or red clover, with
which it must ordinarily compete in agricultural use.
MEDICAGO ORBICULARIS (L.) ALL, (BUTTON CLOVER).
(Pl. VII, fig. 1.)
Stems procumbent, 10 to 80 cm. (4 to 32 inches) long, sparingly pubescent:
leaflets oval to obovate, sometimes truncate at the base, sparingly pubescent
on both sides, up to 15 mm. (five-eighths inch) wide and 20 mm. (seven-
267
a8 NONPERENNIAL MEDICAGOS.,
eighths inch) long, rounded at the apex, the margin toothed nearly to the
pase, the leafstalks 1 to 10 cm. (three-eighths to 4 inches) long; stipules with
slender teeth about 2 to 3 mm. (one-sixteenth to one-eighth inch) long; flowers
4 to 5 mm. (about three-sixteenths inch) long, in pairs, on an axillary peduncle,
the stigma slightly exposed when tripped; pods papery, straw colored, netted
veined, 18 to 20 mm. (about three-fourths inch) in diameter, twisted spirally
into 4 to 6 thin, flattened turns, the margin often recurved, the central wind-
ing largest, the others gradually decreasing in size; seed yellowish brown,
obovoid, flattened, about 2} to 3 mm. (one-eighth inch) long, the surface
minutely papillose, the radicle as long as the seed.
Distribution: France, Spain, and Algeria; thence east to Persia. In the
United States it has been reported growing spontaneously only in Alabama and
southern California.
This species was received from Algeria and from Brunswick, Germany, under
S. P. I. Nos. 10725 and 16876, respectively. It has been tested most extensively
at Chico and other places in California, but has also been tried in several
localities in the South Atlantic and Gulf Coast States. It promises well for
pastures in California, but has not been sufficiently tested in the Southern States
to determine its value there. In general its growth is about like the spotted
and toothed bur clovers, but in California it yields very much more seed than
either of these species. Its large spineless burs and heavy yields of seed make
it superior to common species for pasturage. It matures earlier and is affected
much less by the clover-seed chalcis than the toothed or spotted bur cloyers.
MEDICAGO ORBICULARIS MICROCARPA ROUY AND FOUC,
(Pl. VII, fig. 2.)
' This subspecies differs from typical Medicago orbicularis only in having uni-
formly smaller pods, varying from 8 to 12 mm. (one-fourth to one-half inch) in
diameter, having the same season of growth.
Distribution: Same as for the species.
This importation was from near Oued Smaar, Algeria, under 8S. P. I. No.
7738. Tested in California and in the Southern States in comparison with the
species it shows no difference in agronomic value.
MEDICAGO ORBICULARIS MARGINATA (WILLD.) BENTH.
(Pl, VII, fig. 2.)
This subspecies differs from typical Medicago orbicularis in having pods with
looser windings, all of the same diameter, and the margins always straight,
never recurved. .
Distribution: Same as for the species.
This importation was from Karlsruhe, Germany, under S. P. I. No. 16265, and
has been tested at Chico, Cal. It makes less growth than the species proper,
yields very much less, and therefore is of much less value.
MEDICAGO SCUTELLATA (L.) WILLD. (SNAIL CLOVER).
(Pl. VI, fig. 2.)
Plants densely glandular pubescent throughout; stems procumbent, 10 to 75
em. (4 to 30 inches) long; leaves oval, oblong, or obovate, rarely broadly
cuneate at the base; leaflets up to 15 mm. (five-eighths inch) wide, and 25 mm.
267
me f=
a
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. Yip PLATE VII.
Fig. 1.—PODS AND SEEDS OF MEDICAGO ORBICULARIS, SHOWING VENATION AND WIND-
INGS OF PODS AND MARKINGS OF SEED COATS.
(Enlarged 2 diameters. )
Fic. 2.—PoDs OF TWo SUBSPECIES OF MEDICAGO ORBICULARIS, MARGINATA (UPPER Row)
AND MICROCARPA (LOWER ROW), SHOWING VARIATION IN WINDINGS AND SIZE.
(Enlarged 2 diameters. )
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE VIII.
Fic. 1.—PODS AND SEEDS OF MEDICAGO RUGOSA, SHOWING VENATION AND WINDINGS
OF PODS AND CHARACTERISTICALLY NOTCHED SEEDS.
(Enlarged 2 diameters.)
Fic. 2.—Pops AND SEEDS OF MEDICAGO TUBERCULATA, SHOWING VARIATION OF WIND-
INGS AND SHORT TUBERCULAR SPINES OF PODS AND CHARACTERISTIC NOTCHES IN
SEEDS.
(Enlarged 2 diameters.)
DISTRIBUTION AND DESCRIPTION OF SPECIES. 29
(1 inch) long, coarsely and sharply toothed nearly to the base, the apex obtuse
to acute, the leafstalk rarely much longer than the leaf; stipules up to 4 mm.
(three-sixteenths inch) wide, and 10 mm. (three-eighths inch) long, sparingly
toothed; flowers about 7 mm. (one-fourth inch) long, in clusters of two, the
stigma exposed when tripped; pods 12 to 15 mm. (one-half to five-eighths inch)
in diameter, straw colored, netted veined, with 5 to 8 thin, spiral, cup-shaped
windings; seed kidney shaped, yellow, 5 to 54 mm. (about three-sixteenths
inch) long, the radicle half the length of the seed.
Distribution: Spain and Algeria to Asia Minor.
This species was received under S. P. I. Nos. 9747 from Madrid, Spain;
16267 from Karlsruhe, Germany; and 16877 from Brunswick, Germany. It
has been grown at Chico and other points in California and at several places
in the Southern States. It matures earlier than any of the other species tested,
and makes less growth than the common spotted and toothed bur clovers. In
comparison with others it makes a heavy yield of seed under California condi-
tions, but has done little in tests in the Southern States. On account of its large.
smooth burs, good yield of seed, and early maturity it is valuable for pasturage
in California. The seeds are little attacked by the clover-seed chalcis,
MEDICAGO RUGOSA DESR.
(Pl. VIII, fig. 1.)
Plants sparingly glandular pubescent throughout, except on the upper surface
of the leaves; stems decumbent, 10 to 60 cm. (4 to 24 inches) long; leaflets up
to 15 mm. (five-eighths inch) wide and 20 mm. (seven-eighths inch) long,
broadly truncate and entire at the base, the apex rounded or retuse, sharply
toothed, the leafstalk up to 5 cm. (2 inches) long; flowers about 5 mm. (three-
eighths inch) long, in clusters of two, the stigma slightly exposed when tripped;
pods 7 to 8 mm. (one-fourth to five-sixteenths inch) in diameter, the windings
23 to 4, somewhat inflated, with conspicuous, radiating, marginal strie; seed
kidney shaped, yellow, 3 to 4 mm. (about one-eighth inch) long.
Distribution: Syria, Mesopotamia, and Palestine.
This species was received from Madrid, Spain, under S. P. I. No. 19442. It
has been tried only at Chico, Cal., makes much less growth than the spotted
or toothed bur clovers, and yields considerably less seed. Its season is prac-
tically the same as the spotted or toothed bur clovers. It is little attacked
by the clover-seed chalcis. The absence of spines makes it desirable for further
testing.
MEDICAGO TUBERCULATA (RETZ.) WILLD.
(Pl. VIII, fig. 2.)
Stem procumbent, 10 to 60 cm. (4 to 24 inches) long, sparingly villous, with
brownish hairs; leaflets pubescent on both sides, obovate to ovate, up to 15
mm. (fiveeighths inch) wide, and 24 mm. (three-fourths inch) long, the apex
rounded to acute, closely toothed, fhe base truncate, the terminal pedicels three
to five times longer than the lateral; stipules not deeply toothed; flowers about
5 mm. (three-sixteenths inch) long, in clusters of 5, the stigma not exposed when
tripped; pod 6 to 74 (one-fourth to five-sixteenths inch) in diameter, twisted
spirally into 4 to 5 turns, a few radiating veins near the center of the turn
surrounded by a smooth band bordered by a vein parallel to the dorsal suture.
The parallel vein and dorsal suture are connected by numerous radiating veins,
these are usually swollen at the base when mature. Seed kidney shaped, yellow,
about 4 mm. (one-eighth inch) long, radicle nearly half as long as the seed.
Distribution: France to Algeria; thence east to Syria.
267
80 NONPERENNIAL MEDICAGOS.
This species was received from the Royal Botanic Gardens, Dublin, under
F. C. I. No. 9229 and S. P. I. No. 17783. It has been grown only at Chico, Cal.
Its season is practically the same as the spotted or toothed bur clovers. but it
makes much less growth and yields little seed. It is worthy of further testing
on account of its spineless burs.
MEDICAGO TURBINATA (L.) ALL,
(Pl. IX, fig. 1.)
Plant more or less densely pubescent throughout; stems procumbent, 10 to
60 cm. (4 to 24 inches) long; leaflets up to 16 mm. (five-eighths inch) wide and
25 mm. (1 inch) long, the leafstalk not much longer than the leaf, the stalk of
the terminal leaflet five to eight times longer than the lateral; stipules not deeply
toothed; flowers about 5 mm. (three-sixteenths inch) long, single, or rarely in
clusters of two, the stigma exposed when tripped; pod 7 to 8 mm. (one-fourth
to five-sixteenths inch) in diameter, twisted spirally into about 5 windings,
which are smooth, woody, and sparingly covered with short, stiff, tubercular-
pointed spines; seed kidney shaped, light yellow, 5 to 6 mm. (three-sixteenths
to one-fourth inch) long. Pods without spines sometimes appear in dry seasons.
Distribution: Portugal and Algeria to Asia Minor.
This importation is represented by S. P. I. Nos. 19447 and 19449, both probably
from the Madrid Botanical Gardens. It has been grown at Chico and other
places in Califurnia. Its season of -maturing is about the same as or a little
later than toothed bur clover, but its growth is not as great, and its hard,
woody, spiny pod makes it less desirable for pasture. It is little affected by
the clover-seed chalcis.
MEDICAGO MURICATA (L.) ALL.
(Pl. IX, fig. 2.)
Plant pubescent throughout; stems procumbent, 10 to 50 cm. (4 to 20 inches)
long; leaflets up to 14 mm. (nine-sixteenths iuch) wide and 18 mm. (three-
fourths inch) long, the leafstalks about as long as the leaves, the stalk of the
terminal leaflet five to eight times longer than the lateral; stipules not deeply
toothed; flowers in twos, about 5 mm. long, the stigma exposed when tripped;
pod 6 to 7 mm. (about one-fourth inch) in diameter, spirally twisted into five
to six windings, the lateral veins parallel to the dorsal suture, bearing nearly
opposite, stiff, sharp spines as long as the diameter of the windings, the edges
of the windings partly obscured by the slightly interlocking prickles; seed
about 3 mm. (one-eighth inch) long, yellow. somewhat kidney shaped or angular
along the back, the radicle half as long as the seed and the end turned up so
as to form a small beak near the hilum.
Distribution: Canary Islands, both shores of the Mediterranean, and east to
Syria and Egypt.
This species was received from Madrid, Spain, under 8S. P. I. No. 9748. It
has been grown only at Chico, Cal. The season of maturing is the same as
toothed bur clover, but it makes much less growth. Its yield of seed is light,
and its hard burs, with sharp, heavy spines, make it undesirable for pasturage.
MEDICAGO MUREX (L.) ALL.
(Pl. X, fig. 1.)
Stems procumbent, glabrous, 10 to 50 cm. (4 to 20 inches) long; leaflets
glabrous above, pubescent beneath, the leafstalk about as long as the leaf, the
stalk of the terminal leaflet about 5 times longer than the lateral; stipules
267
eee
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE IX.
Fic. 1.—PODS AND SEEDS OF MEDICAGO TURBINATA, SHOWING WINDINGS OF Pops,
SPINY AND SPINELESS PODS, AND SEEDS STRAIGHT AND NOTCHED ON THE UNDER
SIDE.
(Enlarged 2 diameters. )
Fia. 2.—PoDs AND SEEDS OF MEDICAGO MURICATA, SHOWING WINDINGS OF Pops,
SPINY AND NEARLY SPINELESS PODS, AND ANGULAR SEEDS WITH PROMINENCE AT
THE TIP OF THE RADICLE.
(Enlarged 2 diameters. )
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE X.
Fig. 1.—PODS AND SEEDS OF MEDICAGO MUREX, SHOWING VARIATION OF PODS OFTEN
FOUND ON ONE PLANT.
(Enlarged 2 diameters. )
Fic. 2.—PoDS AND SEEDS OF MEDICAGO RIGIDULA, SHOWING WINDINGS OF Pops, VENA-
TION OF PODS HIDDEN BY. PUBESCENCE, AND PROMINENCE AT THE TIP OF THE
RADICLE
(Enlarged 2 diameters. )
ee
DISTRIBUTION AND DESCRIPTION OF SPECIES. 31
rather deeply toothed, the teeth 3 to 4 mm. (one-eighth inch) long; flowers in
twos, about 4 mm. (three-sixteenths inch) long, the stigma not exposed when
tripped. The pods are very similar in size and appearance to those of the
preceding species, but have 7 to 9 turns, and the spines are more erect and do
not interlock. Pods with few or no spines are produced in dry weather. The
seeds are kidney shaped, about 3 to 4 mm. (one-eighth inch) long, and the
radicle is nearly half the length of the seed.
Distribution: France, Italy, Turkey, and Algeria.
This species is represented by F. C. I. No. 0147, from Jamaica, and S. P. I.
No. 16875, from Brunswick, Germany. It has been tested only at Chico, Cal. It
has practically the same season as toothed bur clover, but makes much less
growth. Its spiny burs make it objectionable for pasturage,
MEDICAGO RIGIDULA (L.) DESR.
(PL xX. fe 2.)
Plants pubescent throughout; stems procumbent, 10 to 50 cm. (4 to 20 inches)
long; leaflets up to 12 mm. (one-half inch) wide and 24 mm. (1 inch) long, the
leafstalk often equaling the leaves, but mostly shorter, the stalk of the terminal
leaflet 3 to 4 times longer than the lateral; stipules not deeply toothed; flowers
in twos, about 5 mm. (three-sixteenths, inch) long, the stigma exposed when
tripped; pods 7 to 8 mm. (about five-sixteenths inch) in diameter, similar to
the preceding species, the windings not so thick and covered with a fine pubes-
cence, the spines somewhat hooked at the tips; seed about 4 mm. (five thirty-
seconds inch) long, yellow, kidney shaped, the radicle about half the length of
the seed, the tips slightly raised.
Distribution: France and Spain; thefice east to the Caucasus, Persia, and
Kgypt.
This species was received from Madrid, Spain, under F. C. I. Nos. 0373 and
0377, and from Strasburg, Germany, as S. P. I. No. 16288. It has been tested
only at Chico, Cal. It makes much less growth than the toothed or spotted bur
clovers and yields little seed. Its comparatively hard pod and stiff spines make
it less desirable for pasturage.
MEDICAGO CILIARIS (L.) ALL.
(Pl. XI, fig. 1.)
Stems decumbent, glabrous, 10 to 100 cm. (4 to 40 inches) long; leaflets up
to 17 mm. (five-eighths inch) wide and 30 mm. (13 inches) long, pubescent be-
neath, the leafstalks slightly pubescent, about equaling the leaves, the stalk of
the terminal leaflet 3 to 5 times longer than the lateral; stipules not deeply
toothed; flowers about 7 mm. (five-sixteenths inch) long, in twos, the stigma
exposed when tripped; pod pubescent, 7 to 12 mm. (five-sixteenths to one-half
inch) in diameter, 10 to 22 mm. (three-eighths to seven-eighths inch) long, 7 to
10 spiral windings thickly covered with stiff, somewhat interlocking spines
about 2 to 3 mm. (one-eighth inch) long; seed 5 to 6 mm. (three-sixteenths to
one-fourth inch) long, kidney shaped, black.
Distribution: France to Madeira; thence east to Asia Minor and Mesopo-
tamia.
This species includes S. P. I.’ Nos. 7742, from Oued Smaar, Algeria, and 9747
and 19435, from Madrid, Spain. It has been tested at Chico and other points in
California. It has practically the same season as toothed or spotted bur clover
and makes equally good growth. Its very spiny pods make it less desirable for
_ pasturage, though it produces a comparatively good crop of seed.
267
32 NONPERENNIAL MEDICAGOS.
MEDICAGO ECHINUS DC. (CALVARY CLOVER).*
(Pl. XJ, fig. 2.)
Stems procumbent, glabrous, 10 to 100 cm. (4 to 40 inches) long; leaflets up
to 15 mm. (five-eighths inch) wide and 24 mm. (1 inch) long, pubescent be
neath or only along the midrib, usually marked with a small reddish spot in the
center of each leaflet, the leafstalk about as long as the leaf, the stalk of the
terminal leaflet 3 to 5 times larger than the lateral; stipules not deeply toothed ;
flowers about 7 mm. (five-sixteenths inch) long, in clusters of 6, the stigma
exposed when tripped; pod ovoid to spheroid, glabrous, 10 to 15 mm. (three-
eighths to five-eighths inch) wide, 15 to 20 mm. (five-eighths to three-fourths
inch) long, 7 to 9 spifal windings, thickly covered with slender, rigid, closely
interlocking spines 5 to 7 mm. (about one-fourth inch) long; seed 5 to 6 mm.
(three-sixteenths to one-fourth inch) long, kidney shaped, black.
Limited observation indicates that it is essential that the stigma be tripped
as in alfalfa before the seed will set, and that crossing is advantageous to
seed setting.”
Distribution: Spain and the Canary Islands; thence east to Italy and Algeria.
This species was selected from S. P. I. No. 7742, from near Oued Smaar,
Algeria. It has been tested only at Chico, Cal. It has practically the same
season as the toothed and spotted bur clovers and makes equally as good
growth. Its very spiny pods make it less desirable for pasturage, though it
produces a comparatively good crop of seed.
The subspecies Medicago echinus variegata (Urban) Ricker (M. interterta
echinus variegata Urban) differs from typical M. echinus in having a large, tri-
angular, dark-reddish spot extending from the base to near the middle of the
leaflet. The stems are less decumbent than those of the typical form, and the
season of maturing is a little later. The name is a new trinomial.
Distribution: Same as for the species.
This importation was received from Brunswick, Germany, under S. P. I. No.
16874, and has the same agronomic value as the species. It perhaps can be
used to advantage in California as an ornamental plant for winter border or
bedding work.
MEDICAGO ARABICA (L.) ALL. (SPOTTED BUR CLOVER).
(Pl. XII, fig. 1.)
Stems procumbent, pubescent, 10 to 100 cm. (4 to 40 inches) long; leaflets
up to 22 mm. (seven-eighths inch) wide, 27 mm. (17s inches) long, pubescent
beneath, a dark-red spot in the center of each leaflet, the leafstalk often 4 to 5
times longer than the leaf, the stalk of the terminal leaflet not more than 2 to 3
times longer than the lateral; stipules not deeply toothed; flowers 4 to 5 mm.
(about three-sixteenths inch) long, in clusters of 5 to 10; pods 34 to 5 mm.
(one-eighth to three-sixteenths inch) in diameter, rather soft, twisted into
83 to 5 spiral windings, the edges bearing numerous interlocking grooved spines
about as long as the width of a winding, the veins inconspic:ious; seed about
24 mm. (three thirty-seconds inch) long, kidney shaped, the radicle somewhat
1 Also called ‘“‘ crown of thorns,” the name being derived from the suggested resem-
blance of one of the windings (PI. XI, fig. 2).
2 Proceedings, Cambridge Philosophical Society, vol. 8, 1894, pp. 142-143. Bulletins,
Kansas Agricultural Experiment Station: No. 151, 1907, p. 101; No. 155, 1908, p. 319.
Circular 24, Bureau of Plant Industry, 1909, p. 8.
267
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE Xl.
Fig. 1.—PODS AND SEEDS OF MEDICAGO CILIARIS, SHOWING VARIATION IN SHAPE OF
PODS, CHARACTER OF WINDINGS, HAIRY SPINES, AND NOTCHED SEED.
(Enlarged 2 diameters. )
Fic. 2.—PODS AND SEEDS OF MEDICAGO ECHINUS, SHOWING INTERLOCKING SPINES, A
SINGLE WINDING (LIKE A CROWN OF THORNS), AND NOTCHED SEED.
(Enlarged 2 diameters. )
Bul. 267, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE’ Xd
Fic. 1.—PODS AND SEEDS OF MEDICAGO ARABICA (UPPER ROW) AND ITS SUBSPECIES
INERMIS (LOWER ROW), SHOWING CHARACTER OF WINDINGS, LACK OF VENATION,
AND PROMINENCE AT THE TIP OF THE RADICLE.
(Enlarged 2 diameters.)
Fic. 2.—Pops AND SEEDS OF MEDICAGO HISPIDA (UPPER ROW) AND ITS SUBSPECIES
NIGRA (LOWER ROW), SHOWING VENATION OF Pops, DIFFERENCE IN NUMBER OF
WINDINGS, LENGTH OF SPINES, AND SHAPE AND SIZE OF SEEDS.
(Enlarged 2 diameters. )
DISTRIBUTION AND DESCRIPTION OF SPECIES. 33
more than half the length of the seed, the end slightly raised, forming a small
beak near the hilum.
Distribution: Ireland to Algeria; thence east to Asia Minor. Established
throughout the Atlantic, Gulf, and California coasts and extending rapidly into
the interior States. It is reported from Colorado.
This has been extensively tested in the South Atlantic and Guif Coast States
under §S. P. I. Nos. 16218, 21550, 23661, and 25878, all from American sources.
It is common throughout this region, and on account of its hardiness has
proved the best bur clover for the region designated. It also does well in
California, where the spiny form is common. Its season for maturing is
medium as compared with other species. It is usnally in bloom from early
until late spring. Wherever it does well it affords a large amount of pasturage
both in the green and the dry state.
MEDICAGO ARABICA INERMIS RICKER (NEW SUBSPECIES).
(Pl. XII, fig. 1.)
This subspecies differs from typical Medicago arabica only in the absence of
spines on the pod. Many seeds of this subspecies have been plan‘ed at Chico.
Cal. In every instance they have produced pods true to type and without spines.
Distribution: Seed received without definite statement of locality (S. P. I.
No. 23284) from Mr. José D. Husbands, of Santiago, Chile. The type specimen
grown from these seeds and collected by Mr. Roland McKee at Chico, Cal.,
June i, 1910, has been deposited in the United States National Herbarium and a
duplicate in the Economic Herbarium of the United States Department of
Agriculture.
This importation has been grown only at Chico, Cal. In growth and season
it is the same as typical Medicago arabica. On account of its spineless pods it
gives promise of being especially serviceable in the Middle Atlantic and Gulf
Coast States, where spotted bur clover in particular does well.
MEDICAGO HISPIDA GAERTN. ( TOOTHED BUR CLOVER).
(Synonym, UM. hispida lappacea (Desr.) Urban.)
Crip ZL fig. 2.)
Stems decumbent, glabrous, 10 to 110 cm. (4 to 44 inches) long, and up to
38 mm. (one-eighth inch) in diameter; leaflets broadly obovate to obcordate,
glabrous above, sparingly pubescent beneath, up to 22 mm. (seven-eighths
inch) wide and 28 mm. (1% inches) long, often containing very small scat-
tered whitish and dark-red spots, which disappear with age or drying, the
apex finely’ toothed and emarginate, the base interruptedly toothed to entire,
the stalk of the lateral leaflets very short, the terminal leaflet about five times
longer than the lateral; stipules with linear teeth up to 3 mm. (one-eighth inch)
long; flowers 4 to 5 mm. (one-eighth to three-sixteenths inch) long, in loose
clusters of 6 to 9 on axillary peduncles, the stigma not exposed when tripped;
pods netted veined, 7 to 10 mm. (one-fourth to three-eighths inch) in diameter,
twisted spirally into 13 to 4 windings with a conspicuous vein on each side of
the winding parallel to the dorsal suture, and a double row of nearly opposite,
stiff, erect, or slightly divergent spines connecting the dorsal suture and its
parallel vein, the length of the spines being from one-half to full width of the
windings; seed from light to brownish yellow, about 3 mm. (one-eighth inch)
long, kidney shaped, the radicle half the length of the seed.
267
34 NONPERENNIAL MEDICAGOS,
Distribution: The species and all its subspecies are natives of the northern
Mediterranean regions, and are now found throughout the region from Spain
to southern Germany and east to Centra] Asia and India. The species has been
introduced extensively in Chile and Argentina and is becoming widely dis-
tributed in California.
This species was received under F. C. I. No. 0301 from Cambridge, England,
and under S. P. I. No. 9736 from Madrid, Spain. It occurs commonly in Cali-
fornia, and has been grown in comparative tests at Chico and other places
in that State. It makes a good growth and is one of the best of the bur
clovers in that respect. Its season for maturing is medium as compared with
the other species, being in bloom in California from March until June, though
in damp places it is found in bloom throughout the year. It is one of the forms
of toothed bur clover already discussed (p. 8). Where it does well it affords
a large amount of pasturage, either green or dry. The seed of this species and
all its subspecies are badly attacked by the clover-seed chalcis. On account
of its spiny burs it is less desirable than the spineless forms for pasturage.
MEDICAGO HISPIDA CONFINIS (KOCH) BURNAT.
(Pl. XIII, fig. 1.)
This subspecies differs from typical Medicago hispida in the general absence of
leaf markings, in having rarely a small spot at the base of the leaf, and in
having 14 to 34 windings and no spines. Short veins connecting the dorsal
suture and lateral veins replace the spines.
Distribution: Same as for the species.
This importation was received under F. C. I. No. 0309 from Berlin, Germany,
and under S. P. I. No. 9737 from Madrid, Spain. It has been tested at Chico
and other places in California and in several localities in the Southern States.
It has the same season as the species proper. It makes a good growth and is one
of the best of the bur clovers in this respect. Its pods are spineless and, for this
reason, it is especially desirable for pasturage. This form of the toothed bur
clover is more fully discussed elsewhere (p. 20).
MEDICAGO HISPIDA RETICULATA (BENTH.) URBAN.
(Pl. XIII, fig. 1.)
This subspecies differs from Medicago hispida confinis only in having a pod
with five windings. It has been occasionally found as a mixture with other
species tested at Chico, Cal. Its growth and season are the same as the typical
M. hispida. On account of its spineless bur it is desirable for pasturage.
Distribution: Same as for the species proper.
MEDICAGO HISPIDA APICULATA (WILLD.) URBAN,
(Pl. XIII, fig. 1.)
This subspecies differs from Medicago hispida reticulata only in the presence
of short spines about as long as the thickness of the windings of the pod.
It was received under F. C. T. Nos. 0266 and 0372 from Madrid, Spain, and under
S. P. I. Nos. 16873 from Brunswick, Germany, and 19431 and 19434 from
Madrid, Spain. It has been tested at Chico and other places in California. Its
growth and season are the same as the species. It is preferable for pasturage
to the more spiny form.
Distribution: Same as for the species proper.
267
Bul. 267, Bureau of Plant Indusiry, U. S. Dept. of Agriculture. PLATE XIII.
4
u
Fic. 1.—PODS OF THREE SUBSPECIES OF MEDICAGO HISPIDA, SHOWING DIFFERENCES
IN WINDINGS, VENATION, ABSENCE OF SPINES, ETC.; ALSO SLIGHTLY NOTCHED
SEEDS OF ONE SUBSPECIES.
Upper row, M. hispida conjfinis; middle row, M. hispida reticulata; lower row, M. hispida apicu-
lata. (Enlarged 2 diameters. )
/
FiG. 2.—PODS AND SEEDS OF TWo SUBSPECIES OF MEDICAGO HISPIDA, SHOWING DIFFER-
ENCES IN WINDINGS, IN LENGTH OF SPINES, AND IN SIZE AND SHAPE OF SEEDS.
Upper row, M. hispida denticulata; lower row, M. hispida terebellum. (Enlarged 2 diameters.)
FURTHER WORK PLANNED. a0
MEDICAGO HISPIDA DENTICULATA (WILLD.) URBAN.
(PI. XIII, fig. 2.)
This subspecies differs from Medicago hispida apiculata in having longer
spines, about as long as half the width of the windings of the pod. This form
has been received from various sources under F. C. I. Nos. 0149, 0151, 0262,
0271, 0272, 0273, 0280, 0874, 0382, 0884, and S. P. I. Nos. 19444, 19450, 19452,
19458, 19455, 20715, 22649, 24596.
This form is the most common of the toothed bur clovers already discussed.
Its growth and season are the same as the species. On account of its spiny
burs it is less desirable for pasturage than the spineless forms.
Distribution: New Brunswick to Florida; thence west to California and
Washington. :
MEDICAGO HISPIDA NIGRA {WILLD.) BURNAT.
(Pl. XII, fig. 2.)
This subspecies differs from typical Medicago hispida in having 4 to 6 wind-
ings of the pod and stout rigid spines equaling or exceeding the width of the
windings.
Distribution: Same as for the species.
This form was received under F. C. I. Nos. 0264, 0269, 0379, and S. P. I. Nos.
9739, 19439, 19448, 26072. All were originally from the Botanic Gardens at
Madrid, Spain.
This importation has been tested at Chico and other places in California
and at severa! localities in the Southern States. Its growth and season are
the same as for the species. On account of its very spiny burs it is less
desirable for pasturage than the spineless forms.
MEDICAGO HISPIDA TEREBELLUM (WILLD.) URBAN.
(Pl. XIII, fig. 2.)
This subspecies differs from Medicago hispida nigra in having pods with
more compact windings, the spines being absent or reduced to rudiments. The
pod is comparatively large and somewhat harder than in the other subspecies.
It was received under F. C. I. No. 0274 from Madrid, Spain, and under S. P. I.
No. 16879 from Brunswick, Germany, and under Nos. 19446 and 19456, probably
also from Madrid. It has been tested at Chico and other places in California
and at several localities in the Southern States. Its growth and season are
the same as for the species proper. On account of the large size of the pod
and the absence of spines, this form is desirable for pasturage. Nos. 0274 and
19446 have matured earlier than the other numbers tested and may be of
greater value, as in a dry season late-maturing varieties do not yield as well.
Distribution: Same as for the species.
FURTHER WORK PLANNED.
The comparative testing of the bur clovers is being continued
with the species discussed in this bulletin, together with a number
of others more recently introduced. The newer introductions will
be tested at various stations throughout the bur-clover sections of
267
36 NONPERENNIAL MEDICAGOS.
the United States, to determine their relative agronomic value in
comparison with the spotted and toothed bur clovers.
It is the intention to bring together as complete a collection of
species as possible. Species and subspecies not discussed in this
bulletin are shown in the list that follows. Species marked with a
star (*) have been recently introduced for this work. Attempts
are being made to obtain the others.
List of nonperennial species and subspecies of Medicago not discussed in this
bulletin.
*Vedicago aschersoniana Urban.
*M.
M.
*VM.
. coronata (L.) Desr.
. daghestanica Rupr.
. disciformis DC.
. galilaea Bois.
blancheana Boiss.
bonarotiana Arcang.
carstiensis Wulf.
M. globosa Presl.
M. orbicularis applanata ( Willd.) A.
and Gr.
M. orbicularis biancae (Tod.) Urban.
M. orbicularis canescens Urban.
M. orbicularis glandulosa Urban.
M. pironae Visiani.
*M. praecor DC.
M. rigidula cinerascens (Jord.) Ur-
ban.
M. granatensis Willd. M. rigidula eriocarpa Rouy and Fouc.
M. hispida microdon (Ehrenb.) M. rigidula morisiana (Jord.) Rouy
*V. hispida reticulata (Benth.) Urban. and Fouc.
. intertexrta (L.) Mill.
M. rigidula timeroyi Boreau.
M. intertexta decandollei (Trin.) Ur- | *M. rotata Boiss.
ban. M. rugosa incisa (Moris) Urban.
M. intertexrta panormitana (Trin.) | *M. soleirolii Duby.
Urban. M. tenoreana Ser.
*VM. laciniata (L.) Mill. *M. tuberculata (Moris) Urban.
*V. litoralis Rhode. M. tuberculata aculeata Moris.
M. litoralis breviseta DC. M. tuberculata apiculata (Bast.) Ur-
M. litoralis pentacycla Urban. ~ ban.
M litoralis tricycla (DC.) Urban. M. tuberculata chiotica Urban.
*M. minima (L.) Grufb. M. turbinata inermis Aschers.
*l1. murex sorrentini (Tin.) Urban. M. turbinata neglecta (Guss.) Urban.
M. muricoleptis Tineo. M. turbinata olivaeformis (Guss.)
M. noeana Boiss. Urban.
M. obscura Retz. M. truncatula Gaertn.
*M. obscura helix (Willd.) Urban. M. truncatula breviaculeata (Moris)
M. obscura lenticularis (Desr.) Ur- Urban.
ban. M. truncatula longeaculeata (Moris)
M. obscura muricata (Willd.) Urban. Urban.
M. obscura tornata (Willd.) Urban. M. truncatula tentaculata (Willd.)
M. orbicularioides Cand. Urban.:
SUMMARY.
The nonperennial species of Medicago consist principally of bur
clovers, mostly annual plants native to the Mediterranean region.
Spotted bur clover (Medicago arabica), toothed bur clover (A.
hispida denticulata), and yellow trefoil (Jf. lupulina) are the only
species now widely distributed in the United States.
267
OO eee
ea
i$
SUMMARY. 37
Spotted bur clover is the species best suited to and most commonly
grown in the Middle Atlantic and Gulf Coast States.
Toothed bur clover is the most common bur clover in California,
but spotted bur clover does equally well there.
Yellow trefoil is quite generally distributed throughout the United
States and makes good growth in practically all sections in which it
occurs.
Yellow trefoil promises to be of value for green manuring, not only
in sections of the eastern United States where crimson clover is
grown, but especially farther north.
The bur clovers are adapted for general use only in sections hav-
ing a very mild winter climate, such as the Southern and Pacific
Coast States. |
Toothed bur clover, spotted bur clover, and yellow trefoil are suited
to varied conditions with regard to soil and moisture.
The stronger growing bur clovers make good pasturage and green-
manuring crops.
The bur clovers sometimes cause bloat in cattle when fed in the
green state.
Yellow trefoil, toothed bur clover, and spotted bur clover seem
to be somewhat unpalatable to stock not used to them.
The feeding value of bur clovers is good, as indicated by general .
experience and also by chemical analyses.
Bur clovers without spines are the most desirable for pasturage.
Medicago hispida confinis is a spineless form of toothed bur clover
that is especially desirable for this reason.
Medicago orbicularis is one of the more recently introduced species
that has large spineless burs and is very promising for pasturage in
California.
Medicago arabica inermis is a new subspecies that has a spineless
bur and is promising for use in sections where spotted bur clover does
well.
Fall seeding of bur clover is necessary for the best results in all
sections having mild winters.
In the Southeastern and Gulf Coast States the first of September
is usually about the right time for seeding.
In California seeding may be done any time during September or
October, but if the land is irrigated before sowing October is best.
In the Eastern and Gulf Coast States it is necessary for the best
results to inoculate the soil in which bur clover is grown for the first
time, but in the Pacific Coast States inoculation is not necessary.
One hundred pounds of burs of either spotted or toothed bur clover
contain 25 to 30 pounds of seed.
267
88 NONPERENNIAL MEDICAGOS,
In seeding any of the species of bur clover 15 pounds of seed
(hulled) per acre should be sown when a thick stand is desired, either
in pastures or cultivated fields.
To handle bur clovers as a seed crop is somewhat expensive, but
not impracticable.
The farm machinery used in harvesting grain and hay crops,
although not suited for handling bur clover, can be used to some
extent. : ;
Most bur clover seed on the market at the present time is obtained
as a waste product from woolen mills, where it has been carried in
the wool, and from screenings of small grains, with which it grows
as a weed in California.
Twenty-three species and subspecies of bur clover have been studied
in connection with the results here presented.
The plants of the various species are very similar in habit of
growth and appearance of stems and leaves.
The burs of the species differ more or less, and their botanical
classification is based largely on these differences.
Most of the species and subspecies studied are made up of definite
types or forms which may be selected and grown as pure strains.
Environmental conditions may cause a wide variation in the burs
of an individual plant, and must be taken into consideration in the
identification of species.
267
prErnonaL COPIES of this publication
may be procured from the SUPERINTEND-
ENT OF DOCUMENTS, Government Printing
Office, Washington, D. C., at 15 cents per copy
Issued January 24, 1913.
U. S. DEPARTMENT OF AGRICULTURE.
BUREAU OF PLANT INDUSTRY—BULLETIN NO, 268,
B. T. GALLOWAY, Chief of Bureau.
;
~ TOBACCO MARKETING IN THE
: UNITED STATES.
a
BY
E. H. MATHEWSON,
Crop Technologist, Tobacco Investigations.
ee SSS SA, ~
ci \S wo
D> PS Mo ’ : Pad
Ys p'aer |
P i age
WASHINGTON:
GOVERNMENT PRINTING OFFICE.
19138.
Wi heey
a ae
. H. Mathewson and G. W. Harris, Crop Technologists.
. A, Allard and C. L. Foubert, Scientific Assistants.
. M. East, E. K. Wibshman, W. W. Green, R. P. Cocke, B. G. Anau True Houser, and
BUREAU OF PLANT INDUSTRY.
Chief of Bureau, BEVERLY T. GALLOWAY.
Assistant Chief of Bureau, WILLIAM A. TAYLOR.
Editor, J. E. ROCKWELL,
Chief Clerk, JAMES E. JONEs.
TOBACCO AND PLANT-NUTRITION INVESTIGATIONS.
SCIENTIFIC STAFF.
W. W. Garner, Physiologist in Charge.
G. Beinhart, D. E. Brown, W. M. Lunn, E. G. Moss, and Otto Olson, Assistants. —
F. Scherffius and J. S. Cunningham, Laperts.
W. Bacon, Assistant Physiologist. »
E. Blohm, Special Agent.
G. T. MeNess, Coleerine rT
268
2
LETTER OF TRANSMITTAL.
U.S. Department or AGRICULTURE,
Bureau or Puanr Inpusrry,
OFFICE OF THE CHIEF,
Washington, D. C., September 14, 1912.
Sir: I have the honor to transmit herewith a manuscript entitled
“Tobacco Marketing in the United States,” by Mr. E. H. Mathewson,
Crop Technologist, Office of Tobacco Investigations, and to recom-
mend that it be published as a bulletin of the series of this Bureau.
It is believed that the information which is here brought together
in a systematic and complete form will be of value to the tobacco
interests of the country.
Respectfully, B. T. Gatitoway,
Chief of Bureau.
Hon. James WItLson,
| Secretary of Agriculture.
268
CONTENTS.
Page
RR ea eee ee rare ee Gs ee ch oes Cenk t a ee 7
The Maryland or Baltimore system of marketing .................----------- 8
RET eRLee CT IAGEEG.., 2. aeune cee Sade ooo. Le oS ek a Te ea 8
mmrmmore as 4 TOUACCO-WIAGe CeNTer. 86.22 sl oe ie ee ee cee ce cee eae 12
ie inose-lear tobacco auction-sales system..........--.-.------.--.--ceceeee 13
The markets of Virginia, North Carolina, and South Carolina.........--. 13
Advantages of the loose-leaf tobacco auction system............-...-.--- 17
E Organization of the loose-leaf tobacco trade....................--------- 18
Danville, one of the largest loose-leaf tobacco markets and centers of the
Se aS ts ee ee a a me! Senne ae 19
Other loose-leaf tobacco markets of Virginia, North Carolina, and South
eR ee eee el a Re ys ad ea Sen oe So ae ed Vek 22
Summary of sales in the loose-leaf bright-tobacco markets for twelve years. . 26
Some historic Virginia tobacco trade and manufacturing centers..........-.--- 27
nner Sheer ea Re Ie I eee ares. oo Soe eat ew oot 27
Eyachbure and Petersburg as market:centers... ........:..2-.2.0.5--222- 30
peeeeminent cl the western marketsis.c25..0./.. 0... .. ee dee ees 31
State regulations of inspections in the western markets .............--.-- 31
| eeeay period. betore. the: Crval Wars .cl stesso. ie RAR 33
New Orleans the principal export point for western tobacco. .......-- 35
Importance of the development of means of transportation.......-.-.-- 35
Beginnings of the more important market centers...........:-.-.----- 36
Development of the western markets since the Civil War.............--- 38
Change of trade from New Orleans to New York. ...............---- 38
Sromin of the Lowsville markets ec ei... Set ok 39
Prose of “the Cincinnati markets yo cote... ose eee fee 39
Saeat-tegt-predics:in Cincinnati sil «sa bt A 41
Grew of the Clarksville:market....g05...-.0-..5 02. Sebi ee 41
Causes for the decline in receipts since the late eighties.............- 44
eee ciatureot the western markets). -...2.).....-..--..-.2- 065 sees secon 45
Development of the direct-buying tendency. ..............--.--------- 45
Influence of the pooling movement on the larger markets.............-.-- 47
ame a tine aarm for loose delivery < tn .c=. .- << =<... - 2.52 -s00--t 20% 48
Loose-leaf tobacco sales from the wagon on the street............--------- 48
The Owensboro loose-leaf tobacco auction system. ............-.--.----- 50
Loose-leaf tobacco auction markets in the West....................---.-- 50
Leal SD (a ean, a a ne aan re iat 51
Success of the loose-leaf tobacco auction system................----- 53
Decline of auction sales of tobacco in hogsheads............-...--------- 53
Louisville and Cincinnati the only distinctive hogshead inspection and ~
De EEE Ud Gopi Ra beats BrP pce > 7 a a hmm ane PS 53
Clarksville, Tenn., the most important dark-tobacco market............-- 54
emririgtieid. Tenmm,, Market... ...50c. ... ere cock ee ee oe bes ce nieee 55
Et WIT RO oe ae so nn we ee woe ce oe aba eee 56
; rr amnrenG MATEO POMS "0. cons... 2 8 ea enn cn ees ene oe 57
‘ Important receiving points in the one-sucker district. .........-.-..--. 57
‘- 268
5
6 CONTENTS.
Present status of the western markets—Continued. . Page. Z
Trade organization and market regulation in the western markets. -.......- 57 ,
Organization of the Louisville tobacco trade.......................-- 58
Warehouse fees... .2.: .-.22 2a. =4- 5 ee ee eT ee 60
Consolidation of warehouse interests:.....2.-.....2-...2.ce.0enueee 61
Private warehouse inspections: ...+.2. 2.25. . ..-¥--- 2-2 seas eee 61
Summary of the receipts at the important hogshead-tobacco markets for ten
YOars. . . f9-- 2 -- + eee eee wee Oe wee ie 61
Method of estimating the average annual production of tobacco in the United
States based on statistics of the Treasury Department. ....................-- 63
ILLUSTRATIONS:
Page
Fic. 1. Loose-leaf tobacco auction warehouse, Danville, Va................... 15
2. Interior of a loose-leaf tobacco auction warehouse, Danville, Va., dur-
ing a sale. .... 0st eee ee 16
3. Interior of a cigar-tobacco auction warehouse, Cincinnati, Ohio, before
a Palen. oc ax cee eee Gn a cin Wia es pin SOM DE Set eee 42
4. Loaded wagons, showing the common method of delivering loose
tobacco, either for private or auction sale, in nearly all export and
manufacturing tobacco districts. ...:.....)./s...2.522 2.00. Se ee 49
5. Wagons waiting to unload during a congestion in the loose-leaf to-
bacco market, Lexington, Ky.............-.- Vue oits aoslee ee 51
6. Exterior of a loose-leaf tobacco auction warehouse, Lexington, Ky. --. 52 $
7. Interior of a large tobacco auction warehouse, Lexington, Ky., during
an off seasons. ; 2c eet. Sas Ss SS ewe CTS oe 52
8. Breaking a hogshead and drawing a sample at a tobacco inspection, b
Louisville, Ky: . ag te 27 , 072 1,988 29,060 1,324.39
cee only Maryland tobacco, but the reclamations on the eastern Ohio type were very
The total number of hogsheads of Maryland tobacco received in
this period was 472,220, an average of 31,481 yearly. The reclama-
tions amounted to $13,602.10, an average of $906.81 yearly. The
value of the average hogshead of Maryland tobacco in this period
has been probably something less than $50. The value of the
reclamations, therefore, are equal to the value of only 18 or 20 hogs-
heads of tobacco annually, which is in the ratio of about 1 to 1,500.
Of course, there were reclamations in a greater proportion than 1
hogshead in every 1,500, because probably in no case would more
than a fraction of the total value of the hogshead be allowed in
the reclamation. Reclamation is not usually resorted to except in
extreme cases. As a matter of fact, mixed and careless, though
not necessarily dishonest, packing is much more prevalent than it
should be and is a constant source of complaint from the buyers.
So-called local or home buyers are scatiered through the growing
sections. They buy scattered crops here and there at a round or
268
12 TOBACCO MARKETING IN THE UNITED STATES.
average price and ship to Baltimore, expecting to make a profit by
the transactions.
In 1910 the French Government changed somewhat its system
of buying Maryland tobacco. Purchases are now made direct
through agents, instead of letting the contract to the best competi-
tive bidder for the major part of the requirements, as heretofore.
Maryland tobacco is naturally of a dry and gumless nature.
Conditioning for shipment abroad is done by the growers them-
selves before putting their tobacco on the Baltimore market. P=
oe g's
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 13
accuracy the total annual movement of tobacco through Baltimore.
This places this city very close to the lead among our centers of
tobacco trade in total movement, and it is probably surpassed only
by New York and Louisville.
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM.
THE MARKETS OF VIRGINIA, NORTH CAROLINA, AND SOUTH CAROLINA.
From colonial times until 1877 the sale of leaf tobacco in Virginia
was based on a system of compulsory State inspection by samplers
appointed by the governor. These samplers were appointed to the
numerous warehouses located at the various central market points
in the State. Tobacco for export was subject to seizure if found in
transit out of the State without the seal of inspection. By a State
law passed in 1844 tobacco that came from the West—that is, from
Kentucky, Tennessee, or Ohio—and which was inspected in Virginia
was branded “ Western,” as tobacco from Maryland was branded
“Maryland,” in order to prevent, so far as possible, their sale as
Virginia leaf, which was in greater favor in both the domestic and
the foreign markets.
This compulsory system of State inspection, although of great
advantage in the colonial period, when the export trade was in its
infancy, had long since outgrown its usefulness and had become so
contaminated with politics in the appointment of the samplers that
it was no longer satisfactory to the tobacco trade. The farmers, how-
ever, feared a change to a private inspection system, and several
vigorous legislative campaigns were necessary before the final over-
throw of the system was accomplished in 1877 by the aid of the
legislature.
The law as revised still provided for the appointment of State
samplers and for the official inspection of all prized hogshead tobacco
belonging to farmers, except as removed from the package and sold
loose, but under somewhat modified restrictions permitting private
inspections. Although the modified laws for the State inspection
of tobacco still stand on the statute books of Virginia, they are as a
matter of fact practically a dead letter, because private inspection
has completely superseded State inspection of prized tobacco,
Meanwhile the practice of selling tobacco loose at auction had
started on a small scale in some of the market towns, particularly
of southern Virginia and North Carolina. The act of 1877 gave
an added impetus to this method of selling by changing the law so as
to permit the sale of loose tobacco on the warehouse floor without
inspection. So far as the sales of loose tobacco are concerned the
Virginia law merely requires the weigher to take a prescribed form
of oath to keep the warehouse scales properly standardized and to
268
14 TOBACCO MARKETING IN THE UNITED STATES. 2
properly weigh the tobacco. The warehouseman must issue a state-
ment to the seller vovering in detail all charges connected with the
sale of the tobacco.
Another agency working for the rapid spread of this system is
the manifest preference of the largest manufacturers and exporters
to purchase leaf tobacco by it rather than by the hogshead and
inspection method.
At the present time, so far as the first-hand sale of tobacco from
farmers is concerned, the loose-leaf tobacco system of selling at public
auction is almost universal in the tobacco districts of Virginia,
North Carolina, and South Carolina.
The system has been brought to a high state of perfection and
efficiency and in its general convenience both to buyers and sellers
surpasses any other method of marketing tobacco or other farm
product that has come under the writer’s observation.
Sales warehouses are located in nearly all the centers of any im-
portance in the tobacco-growing sections of Virginia, North Carolina,
and South Carolina. Most of the important producing counties
have at least one tobacco-market town, and in some cases there are
several markets in a county. The most important exception to this is
in the few counties producing tobacco north of the James River in
Virginia. Except Richmond, there is no tobacco market in Virginia
north of the James.
Most of the larger towns have from two to four sales warehouses
each, and in Danville, Va., which is a very large market, there are
generally six and in some seasons more warehouses open for busi-
ness. The sales warehouses are often substantial brick structures
with a great expanse of floor space, sometimes covering from 25,000
to 30,000 square feet in area, with as few posts or other obstructions
as possible. The roof construction, covering such large expanses
without center supports, is a feat of considerable engineering skill.
An exterior view of the great Acree warehouse in Danville, Va., one
of the largest and most substantial of these loose-leaf auction ware-
houses, is shown in figure 1. In front are some of the farmers’
tobacco wagons lined up after being unloaded.
Competition for business among warehouses is very keen, and con-
veniences and facilities for both sellers and buyers are as complete
as possible with a view of attracting patronage. The effective light-
ing of the larger sales floor is accomplished by skylights and side
windows. Usually the warehouse is so arranged that farmers can
drive inside and unload. Bunks and lounging rooms for the farmers
and stables for the horses are generally provided.
In some sections, as, for example, in the eastern North Carolina
markets, there are basements for grading and tying tobacco, so that
268
:
i
&
é
;
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 15
farmers can have this work done for them at a reasonable price if
they so desire. Generally, however, the farmers themselves grade
and tie up the tobacco in small hands at home, then pack the different
grades one after the other neatly in the wagon bed, and cover the load
with sailcloth, bed blankets, or some other protection from the
weather.
At the warehouse each grade is carefully and neatly piled on a
warehouse truck, carried to the scales and weighed, and a ticket is
attached showing the owner’s name, the number of the lot, the num-
ber of piles or grades in the lot, and the weight of the pile. In ad-
dition there are blank spaces left for filling in at the time of sale,
showing the buyer’s name and the price. Stubs are usually provided
ic. 1.—Loose-leaf tobacco auction warehouse, Danville, Va.
also so that these data may be entered on each part, one to be re-
tained by the warehouseman and the other to go to the purchaser.
Each grade is then placed on the floor in rows, usually allowing about
18 inches each way between piles.
On days when the sales are heavy there will often be from 500 to
1,000 or more piles on the floor at a time. The piles are sometimes
very small, often weighing less than 50 pounds, but sometimes reach-
ing 1,000 pounds or more. They will usually average 150 to 200
pounds each.
The ringing of the warehouse bell gives notice of the beginning of
the sale, which proceeds rapidly, generally at the rate of from 150
to 200 piles per hour. The scene is an interesting one, enhanced by
the quick, snappy crying of bids by the auctioneer, with an occasional
268
16 TOBACCO MARKETING IN THE UNITED STATES.
joke thrown in and a moment of merriment, the entreaties and plead-
ings for a better bid on the part of the sales manager, and the rapid
though sometimes silent bidding by a nod or wink on the part of the
buyers.
The crowd moves slowly along from pile to pile, the ticket
marker immediately putting down the price, the buyer’s name, and
private marks indicating the grade. Close behind the ticket marker
follows a clerk, who calculates the value of the pile and places it on
a slip containing the grower’s name and the number of the pile.
_ In the larger markets, after the sales have continued about half an
hour, these calculations are carried to the office, where other clerks
immediately begin computing the gross and net proceeds, and the
Fic, 2.—Interior of a loose-leaf tobacco auction warehouse, Danville, Va., during
a sale.
payment to the seller, generally in cash, is often begun long before
the sale on the floor is finished and before the warehouseman has
received the purchase price from the buyer. In figure 2 is shown a
section of the floor of Acree’s warehouse in Danville, Va., during the
progress of a sale, which is going on at the farther corner of the room.
The wagons of the buyers are on hand soon after the sale starts,
and the removal of the purchases immediately begins. Large shal-
low baskets are used, which are piled high on the wagons, tier on
tier, to be hauled at once to the buyers’ handling houses. The piles
of tobacco are frequently nearly all removed by the time the sales
are over, and are entirely removed soon afterwards, or at least before
the day is ended.
268
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 17
ADVANTAGES OF THE LOOSE-LEAF TOBACCO AUCTION SYSTEM.
A number of advantages of the loose-leaf tobacco auction system,
both to seller and buyer, are apparent.
The seller is conveniently brought into contact at once with the
buyer under as fair conditions as it is possible to create, so far as a
mere system of selling is concerned. He can see for himself what
his tobacco brings as compared with the general market for similar
grades. The sale is prompt—within the day after he arrives with
his tobacco—and his money is ready immediately in cash or its
equivalent. If he is not satisfied with the prices obtained, he may
reject the sale within a reasonable time limit, and he can then imme-
diately reoffer his tobacco for sale or he may take it home, or to some
other market, or to another warehouse, without charge of any kind.
From the buyer’s standpoint there are also important advantages
connected with the loose-leaf tobacco system. He can look over
carefully, although rather hurriedly, all the tobacco, not merely a
sample, that he buys. In order that tobacco may keep free from
damage by mold, etc., either during storage prior to resale or manu-
facture or during shipment to distant countries, it must be thor-
cughly dried out and conditioned—that is, put into safe keeping ~
order. When sold in hogsheads at first hand, the tobacco comes to
the buyer in all sorts of conditions, good and bad, and some of it
must be redried, a matter to be determined by the purchaser. All
loose tobacco is sold in soft condition, the buyer taking the responsi-
bility of redrying and putting it in safe and uniform keeping order,
which is the most satisfactory way.
Of course there are drawbacks connected with this method of sell-
ing, and it does not give satisfaction in every case, either to the seller
or to the buyer. On the whole, however, it is the most generally
satisfactory method of sale yet devised, and it seems to be able to hold
its ground against the hogshead and inspection method wherever
introduced, so far as first-hand sales from farmers are concerned.
Under this system there is no necessity for official inspection or sam-
pling. The warehouse proprietors in conducting the sales merely
employ an auctioneer and such other clerks, weighers, bookkeepers,
and laborers as are necessary. For this they make a charge to the
seller, which in the case of the larger markets usually consists of
three items, about as follows: (1) An unloading and weighing fee
of 10 cents per 100 pounds; (2) an auction or selling fee at the rate
of 15 cents for each pile of 100 pounds or less and 25 cents for each
pile of more than 100 pounds; (8) a commission of 24 per cent on
the gross proceeds. Some of the markets charge 3 per cent commis-
sion and some have no commission charges whatever. In small mar-
kets the charges are usually less in the aggregate than in the larger
65602°—Bull. 268132
18 TOBACCO MARKETING IN THE UNITED STATES.
markets, as an extra inducement to come. to the smaller market.
Other inducements to patronize the smaller markets are those of
convenience and shorter haul for the immediately surrounding terri-
tory and local pride in building up the business interests of the
home town. Buying interests, however, are usually more fully rep-
resented in the larger markets.
The selling charge for a 1,000-pound load of tobacco divided into
five grades bringing an average of 10 cents per pound, based on the
charges just enumerated, would be as shown below. These charges
are those in effect in most of the “ flue-cured ” tobacco markets and
are the maximum allowed by the State law of North Carolina.
Weighing, at 10 cents per hundred pounds____---_________ $1. 00
Auction fee, at 25 eents-per pile: eee 1.25
Commission, 24 per cenf ‘on $100.2. eee 2.50
Total _ 2.42.21 S25 eee ee 4.75
These charges are considerably higher than they would be if the
same tobacco had been sold in a hogshead by sample, either at the
present scale of prices or those in force under the old State system
of inspection; but there are offsetting advantages, such as lack of
expense for hogsheads and the obtaining of immediate returns.
Furthermore, the tobacco is sold in soft order instead of more or
less dry, which means an increased selling weight because of the
higher moisture content.
ORGANIZATION OF THE LOOSE-LEAF TOBACCO TRADE.
In practically all of the larger loose-leaf tobacco markets the mem-
bers of the tobacco trade, including warehousemen, leaf-tobacco
dealers, and manufacturers, are organized into trade organizations,
which have established market rules or regulations in the interests
of the trade as a whole and in the interests of fair dealing and the
avoidance or settlement of disputes between members.
The supervisor of sales is usually one of the more important
officers of the trade organization. It is this officer’s duty to see that
the rules of the organization are carried out, particularly as regards
the running of the sales, such as looking after the correctness of
scales, the proper spacing of the piles on the floor, the number of
piles sold per hour, and the rotation of sales from warehouse to
warehouse.
The arbitration committee in its function of settling disputes
between members also has most important duties.
The privilege of bidding on or purchasing tobacco at the sales is
usually restricted to members of the organization. The minimum
raise between bids is generally the subject of regulation, as, for
example, a minimum of 10 cents a bid up to $6 per 100 pounds:
25 cents from $6 to $15; 50 cents from $15 to $25; and $1 a bid over
$25 per 100 pounds.
268
eee are wl
ps enn
+
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 19
In most of the larger markets there are from two to four or more
warehouses open for business.. These take turns by mutual agree-
ment with the trade organization as to the order in which the sales
shall be held. The first sale of the day is the most popular with
the farmers, and the schedule is so arranged as to alternate in regu-
lar order among the different warehouses on different days. At
the large markets it is necessary to have two sales going on simul-
taneously, each with its corps of buyers, etc., in order to complete
the day’s work. On the Danville market the sales are run in
triplicate through much of the season.
Competition among warehousemen in the larger market centers for
the farmers’ patronage has been very keen and expensive. This
has resulted in the warehousemen pooling or consolidating their
interests in some of the larger markets and placing the manage-
“ment of all the warehouses in town under.a joint management. Dan-
ville and Lynchburg, Va., are notable examples of this consolidation
of interest and management.
On the other hand, selling charges to the farmer are rather high,
usually amounting to more than 4 per cent of the total value of the
crop, and the profits of the warehousemen have been liberal. This
has led in some instances to a movement toward the joint ownership
and management of the warehouses by the farmers themselves.
The most notable and successful movement of this kind is that
of the Farmers’ Consolidated Warehouse Co., of Greenville, N. C.
This company has established branches in many of the other im-
portant new belt markets, including Wilson, Kinston, and Wil-
liamston in North Carolina and Mullins in South Carolina. It has
also invaded the western field in Maysville, Ky., with the establish-
ment of a loose-leaf market at that point in the fall of 1909. The
farmers of the locality in each case subscribed to a majority of the
stock.
DANVILLE, ONE OF THE LARGEST LOOSE-LEAF TOBACCO MARKETS AND
CENTERS OF THE LEAF-TOBACCO TRADE.
With the development in the production of flue-cured tobacco since
the Civil War, Danville forged rapidly to the front as the pre-
eminent market for the sale of bright leaf, and since the eighties of
the past century for many years ranked as the largest loose-leaf
tobacco market of the world. The first-hand sales at one time ran
over 40,000,000 pounds of loose-leaf tobacco yearly. Other large
markets have also rapidly sprung up in the flue-cured tobacco section,
with which Danville has had to share patronage, although this has
not as yet by any means endangered its preeminence in the seaboard
States as a leaf center. The annual sales of loose-leaf tobacco now
run in the neighborhood of 35,000,000 pounds.
268
20 TOBACCO MARKETING IN THE UNITED STATES.
The tobacco trade of Danville had become of considerable impor-
tance early jn the nineteenth century, and warehouses were established
for the inspection of tobacco under the State system. The panic of
1837, however, struck the town with disastrous effect and its tobacco
trade went to pieces, not to be revived until just before the breaking
out of the Civil War. It was the custom to ship the tobacco produced
in this section to the Lynchburg, Petersburg, and Richmond markets.
In 1858 interest had greatly revived, and Neal & Graves erected a
large wooden building for the sale of leaf tobacco at auction, known
as Neal’s warehouse. The opening of this warehouse is generally
recognized as the beginning of the Danville tobacco market upon its
present basis. In 1860 the legislature estabiished an official inspection
of tobacco at this warehouse. The sales were practically all of loose
tobacco, and the venture proved successful. The growth of Danville
as a city has been intimately connected with its growth as a tobacco
market. In 1850 the population of Danville was 1,760; in 1860 it
was 3,500. The. tobacco trade of Danville was, of course, greatly set
back during the Civil War period, but it immediately revived after
the war was over. The rapid development of the tobacco trade also
meant the rapid development of Danville as a business center. In
1900 the population of Danville was 16,520.
The following record of the yearly sales of loose tobacco on the
warehouse floor from 1869, the earliest date at which reliable records
are available, to 1909 will be of interest as illustrating the growth of
the greatest loose-leaf tobacco market in the world.’ The figures are
tukori from the annual report of Mr. A. B. Carrington, president of
the Danville Tobacco Association.
TABLE IIl.—Record of the yearly sales of loose-leaf tobacco at auction on the
Danville (Va.) market from 1869 to 1909, inclusive.
| Average o ‘ ——
“he Quantity price , uantity price
Year. sold. per 100 Year. sold. per 100
| pounds. pounds,
| Pounds. Pounds.
S008... Tete ee eee 10, 621, 557 $12.95 || 1800..<.-..-- ssid SOR $11.95
rr ES EE GS Yd 13,191, 406 12.00 || 1901s... .2......2.2-.....5) ~ ST 8.87
Ws isd cinta kebeeceae 14,065,639 12,34 }| 290%. 0.2.22 ca Sa 8.23
1 | EES ART RES NS,’ 15,827,846 1} 06: 20h. Bete : 42,050,141 6.46
7 RE ERAS 8.5 MS TS 16,147,715 Tee e ! Aa Se Se . 39, 206,789 7.96
PS PR CAPA | 14,679,421. 9045 |] WMO occa cc cused | 40,160,999 7.79
TOT Darn. as natn a oan 23,466,413 Ch de) Me oS 6.46
BIG. ok vices Wade ee 16,624, 296 19-901 3007 ss. cas cca chet ected ' 49,464,741 7.81
To RR alae, ROME Seat ica 5 27,698,125 8.80 || 1808. .c.--.---co-cca.-d @O;0Oeeee 6.64
OIG os snitch =te emai 26,8 11.01-1|"1900...2....02..--.i. 45.247 | 0 6.74
py a SE PEI aD 83,151,247 11.39 |; 1900- : : Re DS a 3 > | 28,808,846 | 8.75 || 1900-..-2.-.:--.2-.-.--—+4 86,897, 6id 10.62
WO 6.05 cis eee 24,925,076 | 13,22 ,
1Since 1910 the loose-leaf sales in the Lexington, Ky., market have equaled or
exceeded those at Danville.
268
-
i. ee ~ nn ei
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 21
This table shows that the high-water mark in the sales on the Dan-
ville market was reached in 1899, when more than 50,000,000 pounds
were sold. The decline, however, does not indicate a decline in the
crop produced. The real reason is found in the development of a
large number of small markets throughout the producing territory.
Formerly, the Danville sales included much tobacco shipped from
distant points. Now, however, this business is much smaller and the
sales represent almost entirely tobacco brought in on wagons from
the territory immediately surrounding the city and within hauling
distance, usually not exceeding 30 miles.
A marked change has been noted in late years in regard to the
time of marketing tobacco. Farmers formerly delayed marketing
heavily until after the Christmas holidays. Recently, however, there
has been noted a tendency to market early, and the bulk of the crop
is now often marketed in the first few months of the selling season.
In 1908 more than a fourth of the entire crop marketed at Danville
was sold in the month of October.
In the New Belt the crop is sold earlier and more rapidly than in
the Old Belt. In South Carolina the crop is harvested in July and
sales begin immediately; August and September are usually the
heavy months for sales and the market is practically closed by No-
vember 1.
Table IIT shows the monthly sales on the Danville market for the
years 1876, 1890, and 1908, illustrative of this change in time of sell-
ing the crop. The data were kindly furnished by Dibrell Bros.,
Danville, Va.
TABLE III.—Record of the monthly sales, with average price realized, for tobacco
sold on the Danville (Va.) market in 1876, 1890, and 1908.
Sales.
onthe: Crop of 1876. Crop of 1890. | Crop of 1908.
Quantity, | AVeT@8e | Quantity, | AVeT@S" | Quantity. | Average
‘| price ‘| price. : ce
Pounds. Pounds. Pounds.
APE eee Rn PA ye ee 900, 902 $18.01 4,155,750 $11.62 | 10,107,125 $9.77
TTPO ae a 959,711 11.79 3,883,935 10.56 6,401,013 9.77
(eeerarerenyalee ste Ne 107,655 10.80 1,488 , 763 10.01 4,719,917 9.89
1 OLR DHT 5 ey eo 696,448 12.16 2,019, 562 12.05 4,813,471 10.41
pirate re are fe ee ie 1,176,839 14.35 5,479,977 11.62 4,815,587 10.36
01 0 UD as eS ee ee 1,457,988 12.43 4,669,455 11.88 3,761,626 9.62
CLL LLL ss I ee a 1,808,872 12.08 4,551,081 12..59 1,770,595 9.11
SL 5 Dl er 2,562,190 10.75 3,368,621 13.68 785,733 8.68
CTE. 0 ss Dg edn ee 1,960,338 12.30 3,199,692 12.35 327, 503 3.31
AR 5 ee Te 1,664,759 11.04 3,605,113 13.04 @) ()
Oo Gna ask ee 2,044,124 11.94 2-40 tee 11.84 1 613,259 15.46
Oneal ee es 1,221,470 11.58 1,399,617 11.08 1 916,224 18.75
ieee eee oe 16,624,296 12.23 | 40,099,289 11.95 | 39,062,004 9.77
1 Market closed in July. The sales year now begins on Aug. 1 instead of Oct. 1, in response
to this change in time of selling. August and September sales in this column should stand
really at the top, representing the first sales of the 1908 crop, consisting mostly of primings,
and thus accounting for the low average price for these months.
268
yA TOBACCO MARKETING IN THE UNITED STATES.
The commanding importance of Danville in the flue-cured leaf
trade is not adequately measured merely by the pounds of leaf sold
on its warehouse floors. Danville is the one great central receiving
and distributing point for all types of bright leaf as produced in
the Piedmont or western portion of the flue-cured belt, the great
middle district in. which Danville is situated, and the New-Belt
Coastal-Plain section of eastern North Carolina and South Carolina.
In Danville are located the greatest array of sales warehouses, redry-
ing plants, stemmeries, and establishments of tobacco-leaf dealers
and commission merchants of any market in the flue-cured tobacco
belt.
The total movement of tobacco through the Danville market is
greatly in excess of the sales of loose tobacco. Aside from the leaf
sold through the auction warehouses, from ten to fifteen million
pounds of leaf come into this market yearly through the purchases
of dealers or through consignments to them from other markets.
This brings the total volume of the leaf trade of Danville to upward
of 50,000,000 pounds per annum. We have spoken of Danville as one
of the largest loose-leaf tobacco markets in the country. There are
other markets that handle a much larger volume of leaf than Dan-
ville, notably Louisville, Ky., whose annual sales generally run more
than 100,000,000 pounds. A great but uncertain portion of this
larger trade of other markets consists of dealers’ tobacco, 1. e., tobacco
bought up speculatively by country buyers who travel through the
growing districts picking up individual crops here and there. If the
truth could be accurately determined, it might be found that Danville
would be a leading market not only in the sales of loose tobacco but
also in first-hand sales from farmers. Practically all the tobacco sold
loose at the Danville warehouses is first-hand sales direct from
farmers.
OTHER LOOSE-LEAF TOBACCO MARKETS OF VIRGINIA, NORTH CAROLINA,
AND SOUTH CAROLINA.
There were 23 loose-leaf tobacco markets in operation in Virginia
in the sales year 1908-9. One of these, the Richmond market, sold
“sun cured ” tobacco almost exclusively, while another, Amelia, sold
the olive-stemming type exclusively. Twelve markets sold tobacco
of the dark-fired type, either chiefly or exclusively, and nine were
principally or entirely flue-cured or bright tobacco markets.
In North Carolina there were in the same year 45 loose-leaf
tobacco markets which sold the flue-cured type of leaf, 20 being in
the Old-Belt and 25 in the New-Belt section.
In South Carolina during the same period there were 12 markets
in operation, all selling the New-Belt flue-cured type of leaf.
268
nites. ast Lui tatiana aan ga peta
THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM.
for all sales per hundred pounds. The returns for the South
lina markets are for the year 1909, based on the reports made
returns to the respective commissioners of agriculture of those
under the State laws. The price averages for each market in Vi
were furnished through the courtesy of Dibrell Bros., Danville,
South Carolina for the years specified.
VIRGINIA DARK-TOBACCO MARKETS SELLING 1908 CROP.
23
A complete list of these markets is given in Table IV, with the
amount of their first-hand sales for producers and the average price
Caro-
under
the law to the commissioner of agriculture of that State. Those for
Virginia and North Carolina are for the 1908 crop, based on the
States
rginia
and North Carolina were obtained from the different warehousemen,
as determined by the actual sales records kept from day to day, and
Va.
TABLE I1V.—Record of the sales of leaf tobacco and the average prices obtained
at the loose-leaf tobacco auction markets of Virginia, North Carolina, and
Principal type of| Quantity | 4VeTase
Market. tobacco sold first} sold first per'100
hand. hand. pounds.
Pounds
NI ne nS cs eed nage man Gured.2222.025552 9,371,576 $8.06
OTe a dios ote fe aE te a pe nS, ee Olive stemming”__-__-- 829,070 8.50
6 jy TIES 5 3 aaah ae eS Re a oy ae ee ae Dark fired 2222 226 20,178,700 7.20
ater ee ta ee A Se En eS ee COG25 5. —- So ae 6,340,260 7.14
UE SES TEPC cee a a ee lee el ee GOL. 24 ee al Sea 15
OT SS Sp aS ae ee RO ee (alo aa ke = eg a 4,184,208 7.02
RRS TNR Beste een he 2) ee TS a ee | ee Oss eee See eee 3,796,476 7.00
eNESRAuCh REC eer ocr ee SER. eee A ta Soe Ske. (Oe ers ee 2,456,357 7.30
US SAS 5 Se eee Se eae ee eee gos Ne Ts Sere |) ae Cee ee eee 1,904,147 6.73
REPU Teg Sil eee Se A ae eg eee (aaa Set eae cen CROs 1,125,000 6.80
rena ee? tc of) 2 a Se ey NS ri Re aot 687,190 | 6.75
TELE on - Stipe aR SSSR SR eer lr ae es ema tee) Oe 2 a ee ee 400,000 6.75
PII 2s oe oon ten hale ae on "0 ig teen ug Se 51,698 6.00
TESS Hee le a Se ee I eS iy Ret | ee ee ree 56, 543 , 402 T.a2
VIRGINIA OLD-BELT BRIGHT-TOBACCO MARKETS SELLING 1908 CROP.
ee. IE fre bee Pa baw en ee gee Fige eured.......<-.-.- 34,348,914 | $9.80
x BeRRTURESEYAUO TIMES ee hoe oe a ee ee 0 _| 15,556, 807 9.38
nt IEEE Arne Sar. ee me 9 eS ie ke ee joo ohhh ae 3,251,408 9.50
~ ELEN: SS SE Se Se eee eee alien SIMPY SOE eee ee ee, | ae ee Oss eeo 2252 sh Ba OE So 8.75
a 3 TEC iy Re Sete eae 1 ste Ee Se ee | 2,498,485 8.00
a NPR URDM sea. Shee a ee oe a oe Ske pe ou 0 2 ee eee | 2,411,994 8.50
p | (2 TTS SS GR a eas aN sa ae es Somers op A Re Oe oe ee a OOD 8.25
‘ ES pemretn Orne re a Se SB eo ee Oe oe eee LB Tlb08 8.00
RINE IE sant Sees eee a Sok Se cae See oe LL SG ee 821,989 7.64
IE a on ed ee cee ee Lae MOR) eo SL ORE OUS 9.25
= So SSS Soe re ee) |) re 65,769,317 | 9.50
rs NORTH CAROLINA OLD-BELT BRIGHT-TOBACCO MARKETS SELLING 1908 CROP.
‘
me Re WGN eens. f= oe te 8 i cacccasakheee ue Hlge@eured. 2... 2-222: 20,939, 200 $10.23
* (TR st ER a I (ne Riga oF eo 10.87
a OL see 55 22 ee cine ae ere ames Week 25. = LP 6 eeG- ere 12.40
RIE a eee oe oe yt be BS ot Seal ae Ost. oe Se ee Tae 9.46
E EDS) SEES Sa ea era ee es oe) eer ae Cae eS ee 5,279,709 11.28
Roxboro. wes an. DLE SS Sa ee ee ee ee |e eee ee 4,760,322 10.41
=. bore DE Sac eh sa 3, eS ne || Roose 2 Sk Oe Sr ee 10.00
is DE Ts a). OSE ee SO eee ne ee (er ee, eit t A Oo AS AOR SG 9.85
- SLO eae eo OS ee ee a ea PC WA SS ae 2,214,698 9.00
S; Wiener eh riPunrennney ete Se ok Fee at Bee oo ef ime: te es eae ar 9.08
1 Farmville sales only partially reported; should have been over 7,000,000 pounds.
268
24 TOBACCO MARKETING IN THE UNITED STATES.
TasBLe 1V.—lecord of the sales of leaf tobacco, etc.—Continued.
*
‘NORTH CAROLINA OLD-BELT BRIGHT-TOBACCO MARKETS SELLING 1908 CROP—Con.
Quantity | Average
Market. Type. sold first | _ Price
hand. per 100
pounds.
Pounds. ,
Oresdmore. ...- Sa... 255 Sif52 eee Fine curred... -_.-_--.2: 1,732,500 $15.50
Madison. - < Js. ~ Sosa ee 1 ae NE = 1,636,691 9.00
Greensboro: . 22 0222.5 sce eae MG aes sates toc 1,140,811 9.04
Burlington. -...: .. -.22. - 2a eee ee | Ce rene 1,071, 558 9.00
AMOK 2: oe ovale ~-ecdes es eee eee ce ee 1 ph Mee : 15.63
Voungsville:.--222... 4 So. ee eee eee do 2s Be 848,891 10.38
Belelgh.wsc.: ia ook oS ee fe Lt Ww teen, 2s ee ee 497 ,429 11.00
Statesville. .:.-. 2-12-35 Seeger ce [+ oe Sere 286,101 8.50
Lieaksyille-2 i222. ssi0 io Sac a er es eee 5 paar - Sea op 225,718 10.00
Pilotmountain..\..2.-. 22.2020 eee G02 =e ee 205, 456 10.00
Total. . 2... 20-2 oo oes De ee See ee ee el ee 71,785,788 10.00
Wilsons ts.2 22.0 eee LCA Uh 6 eee 16,435,712
Greenville: 222-2. 22.8 oe eee eee O02 eee ee eS , 030,
Kinston... sso Le 3 22. eee Gis 2a sg hee 9,298,021
Rocky Motint:2: 222627. 2 _ ee See ee ele OSs 22 ees es 9,183,146
Marmy ile. 2223's ot 2 ba eo ee eee ede (6 (¢ eS
Goldsboro... 2-2-2 20S eee re (0 ee eee BR Me ae 2,458,458
Smithfield_..--22- 2.2.2. 22: 2 eee See 62222 eee 2,150,618
La Grange. i222. Lee ee ee eee des < "ss as 2,113,604
Hair. Blatt eo a oe <0 (cea aides ae 1,707,774
Robersonvillew: 2... 42=-00 2 2 ee eee ee oho aioe a ie Sa , 563,
Willisimst oni 8 025 2 ee eee ee ee eee no eee eee 1,488,352
Mairmont J. 22S 2 tee eee ee 2 eee ae Ss 0 ar EDE ey PEs
Richlands: 25} ee Se eee Se LO an ote ee 1,385,470
Snow. GM. oes} si, oe. ta Be ae. Ses ene ee Pe Ss Bok SOY Se | 1,284,
ASV Gan 8 ts Pe ee ee ee a eee, ens eee ip eS 5 | 9 Ra Ke i. Sipe oe Ne | 1,181,533
ZSOWMON: 6. eo ee ee ee ce ee 2. SSGe 22 Se eee 950,641
Volos | a fc epee ee ea SE Rm BOSSE 2 Se yee ee EE APL (oye RE ene See wed ge 783,218
Mnield: 5435. Ss we RES eS ee tee Rien eee Be eae 5 (0 Pee oe Oy ine SRE 721,793
"Wierdeall. 2. oO". 2 seo 0 Wee ae ne oe ee tO ee Ce ee 665,
WeATSS Woe. oct Noe ee ees | ee aera Press {, Met eS ete 607,174
Gumaberton 233-2. 2 eae ee ee eee ee eee | cae pao ae oe 454,003
OUOtOR 3s aise sk ed ee eee ee eee ee 22 eS ee al
DUM sso Sone ee Ee See ee eae |e PACS So 349, 592
Muduasy Springs. .<.2-) 2 Sa ee eee dor ee ee 230,291
Olarkton =: cee wk oo ee ee ee ee ee rs (2 Re a eae arora 106,124
Totals £......6 Soh G ccs tere ee de re ee ae ee ic ee 70,433,517
Bee wukeyeekeuerpurypepeese
© 0 “IAT AT AT ATaT~1~T-1-1-1 00 HH HO ~IHMMoOMOoB
_
i—}
Quantity yet
Market. Type. sold first 109 Total value.
: hand. —
pounds
Pounds.
RUINS oe eA 2 seat aes a eee emma Flue cured-_-.-.--..- 6,805,899 $6.89 $468, 982.85
WALID COD eit oe oh wk ee ee ee Caen ened Cnn ck eka. eps ce 7.94 377,045.43
Dike Oly 22-0. = = 02 Sel oats ee ee ee eae 0c 5 4,448, 299 7.97 354,770.61
"THIN ONS VIG: «... < nuviem uciee Geeioewene eee De laasee re | ¢ as eee RS RE DT Ry! 7.68 310,905.23
WiIONGHGGs &. Sacss 2 OE a ee eee WO cnn te et a ee 7.34 166,971.30
1) tins (0) 0 ag ee aaa a) SY SN Dene (ES eh r: (pone es ver figs RR wg 6.54 118, 966.96
Nicholas... 228d ha oO eee ee ro {? ees fees Une 6.66 96,380.91
Kingstree. 42. ot ee ee ead 40... 2.0343. 4|° ae 7.67 108,749.24
Manting’ 4% oh cebu. as dae eee te ee ee ee ee G0 ol eee 6.89 80,316.77
0vig. 2... telah e Seed. See eee eee eee do. St..-223622) . 1, 078480 5.45 58,774.47
TOMI OE. 2 oc he we cee ae eee et ee Eee OO ee eee 1,049,961 5.91 62,068.04
Latta ee ee ee ee ee 5 | 5 ee a 984, 782 7.00 65, 428.83
O60 W AY.cc0.csevar Cates meee eenokeira an ae Cet aae ry Neate BE ace 741,711 6.17 45,791.69
TOtalel cscs See re cueneednst ba cece een] aos bieeee Cee el ee 7.27 | 2,815,168.338
268
’THE LOOSE-LEAF TOBACCO AUCTION-SALES SYSTEM. 25
Of the 13 markets operating in South Carolina in 1909, 7 (Mullins,
Nichols, Loris, Dillon, Latta, Marion, and Conway) were in the
section east of the Pedee River. The combined total of their sales
amounted to 13,875,050 pounds, selling at an average of $6 per 100
pounds, while the 6 markets on the west side of the Pedee sold
17,945,469 pounds, at an average of $7.78 per 100 pounds. Prac-
tically no tobacco is carried across the Pedee by farmers, so that
the sales of the markets on either side of the river closely represent
the tobacco produced on that side. The markets of the western side
of the river, however, do not always make a higher average over
those on the eastern side. As previously stated, the land on the
eastern side is, on the whole, sandier than that on the western side.
The 1909 crop season was particularly unfavorable to sandy-land
tobacco, as compared with that grown on stiffer land. In some
seasons this.condition is reversed.
Sometimes proximity to a body of land producing a type of leaf
above the average in quality will have a material effect in raising
the average price of a market. The averages of Lake City and
Darlington, S. C., can both be explained in this way. The Creed-
moor (N. C.) market is another striking illustration of this point.
Creedmoor draws largely on the famous Dutchville section of Gran-
ville County for the tobacco sold in its warehouses.
The South Carolina law requires that each warehouseman report
the grade of tobacco sold, together with the price. This grade, of
course, can only be approximated, as the warehousemen have no way
of knowing the grade except as a matter of judgment within broad
lines. As reported, however, the 1909 sales of the South Carolina
‘3 markets, which may be of interest, were divided as shown in Table V.
.
& TABLE V.—Record of the sales of tobacco on the loose-leaf tobacco market of
South Carolina by grade and price for the year 1909.
Average
* : price
P Grade. Quantity. per 100
-" pounds.
2 Pounds.
ee ee eee ee 158,386 $12.05
UR Ts oe ee ee ect Se ee 8 ec ek 2,425,101 8.06
anes nnn NCa! eee SD ee eo asa See eee ee ee 24,101,811 7.44
ee ne a peas res Oe Ses se NE - oe eee. 513,746 7.19
DRMinerrmrinemeciunninl edt 258. 8 So ee oe Scud oe eee eee nk 1,212,801 6.00
ENEMA OO COs wees 20 eke on. oe so aa oa San OR eee Ha nek Sel 1,077,141 5.79
7 PL SUT ESSl. oe Ss eee ES ok ee eRe me Sera ek ee 2,322,518 4.52
nn 2a en ae Kt ee 8,997 | 1.06
eo ae ie oe. || 31,820,501 7.27
Evidently most of the warehousemen were inclined to put all the
leaf grades into one class, as leaf. The returns as a whole show
entire lack of a system in reporting the grades.
268
26 TOBACCO MARKETING IN THE UNITED STATES.
The sales by months in the South Carolina markets, 1908 crop,
were as follows:
; Pounds.
July ____ a a ee aa 4, 529, 833
August —__o2 22-2 3522 See 12, 870, 419
September —_ =~. 2-52 ee
October _~.- 223 2 ee eee 3, 976, 807
November ___~. 4) 22222 eee 303, OS5
Total ___ s3. 2" 2 ee ee ee ee ee 31, 820, 519
It is not to be supposed, of course, that these lists of markets are
identical from year to year. The larger markets are generally fix-
tures, but the smaller ones are coming and going from year to year
under the influence of various local causes. .
SUMMARY OF SALES IN THE LOOSE-LEAF BRIGHT-TOBACCO MARKETS FOR
TWELVE YEARS.
The sales of tobacco in the markets of the Old-Belt sections of Vir- -
ginia and North Carolina and in those of the New-Belt sections of east-
ern North Carolina and South Carolina as a whole for the 12 years,
from 1898 to 1909, inclusive, are shown in Table VI. The figures
were kindly furnished by Dibrell Bros., of Danville, Va., from the
returns obtained from warehousemen in all the markets, based on
actual daily transactions. However, they include resales, so in order
to get at the first-hand sales with approximate accuracy, they should
be reduced by about 10 per cent. The correction is unnecessary, of
course, in connection with the -price averages.
.
E
E
;
TABLE VI.—Sales of tobacco with the average price obtained on the Old-Belt and
New-Belt flue-cured markets for 1898 to 1909, inclusive.
Average
Crop. Market. Sales. price per
100 pounds.
Pounds.
1898... | Old Belt (Virginia and North Oarolina)...-2. 22 eee 116,000,000 $6.50
1806... New Belt (eastern North:Garolina) 5. 24020 Jee cease 47,000,000 6.75
1896:..| New Belt (South Oar): - --
4
ol
Y
Dg
«
‘
PRESENT STATUS OF THE WESTERN MARKETS. 51
been greatest in the Burley district, and since 1908 the loose markets
have rapidly multiplied in that section, extending also into the upper
Green River section of the dark one-sucker district, with large sales
warehouses at Glasgow and Bowling Green. In figure 5 a scene in
the warehouse district of Lexington, Ky., is presented, showing an
accumulation of wagons waiting for a chance to unload, the ware-
house floor space being insufficient to hold all the tobacco at a single
sale.
The selling charges in the western loose-leaf tobacco markets are
on the average somewhat lower than in the Virginia and North Caro-
lina markets. Instead of three items of charge, only two are made.
The so-called auction fee is omitted. The charges at Lexington, with
which those of the other western markets are substantially uniform,
are 15 cents per hundredweight and 2 per cent commission.
=.
Loa iy
Fic. 5.—Wagons waiting to unload during a congestion in the loose-leaf tobacco
market, Lexington, Ky.
The warehouses, particularly as established at Lexington, are fully
equal in size and construction to any that can be found in the East.
Some of them have concrete floors and walls, and the immense roof
expanse is often of steel trusses and the framework of real architec-
tural interest.
THE LEXINGTON MARKET.
Lexington, Ky., has already become one of the great loose-leaf to-
bacco auction markets of the country. In the season 1909-10, selling
the 1909 crop, the sales of leaf at Lexington in these auction houses
were fully 20,000,000 pounds. Only Danville, Va., and Winston, N. C.
exceeded Lexington in the volume of such loose sales) Many mem-
bers of the trade confidently predict that it will be only a few years
before Lexington will outdistance all markets, even including Dan-
268
52 TOBACCO MARKETING IN THE UNITED STATES.
ville, and become the greatest loose-leaf tobacco auction market of the
country.!. The railroads have modified their freight charges so that
tobacco can be transported loose for a considerable distance at reason-
able rates.
Fic. 6.—Exterior of a loose-leaf tobacco auction warehouse, Lexington, Ky.
In the sales season of 1909-10 six large warehouses were in opera-
tion at Lexington and a seventh was constructed for 1910-11. A
number of dealers and commission merchants have established head-
quarters there. The trade has been organized under the title of the
47
2 ’
at wv
cm 5c!
Was
|
VAN 25 7
;
e
>L>
Fic, 7.—Interior of a large tobacco auction warehouse, Lexington, Ky., during an
off season.
Lexington ‘Tobacco Association for its better control and supervision
in the best interests of all. Figure 6 shows an exterior and figure
7 an interior view of one of these large loose-leaf tobacco warehouses
at Lexington.
1Sece footnote on p. 20.
|a2
~65
PRESENT STATUS OF THE WESTERN MARKETS. 53
SUCCESS OF THE LOOSE-LEAI TOBACCO AUCTION SYSTEM.
The loose-leaf tobacco auction system of first-hand sales now seems
to have a firm foothold in the West, particularly in the Burley district.
Such markets, on the Virginia plan, are already in operation or in con-
templation in Lexington, Danville, Maysville, Frankfort, Springfield,
Carrollton, Glasgow, Bowling Green, Hopkinsville, and Paducah,
Ky.; Madison, Ind.; Clarksville, Tenn.; Huntington, W. Va.; St.
Joseph, Mo., and elsewhere.
DECLINE OF AUCTION SALES OF TOBACCO IN HOGSHEADS.
Accompanying the development of very unusual conditions, which
have beset the old-established trade methods of the western tobacco
markets, assisted no doubt by generally advancing prices, there has
been a marked tendency to handle an increasing proportion of the
inspected hogshead tobacco by private sale rather than at public
auction. All of the regular markets have exhibited this tendency
strongly and, indeed, with the exception of Louisville, Cincinnati,
and Clarksville, the public auction sales of hogshead tobacco may be
said to be almost a thing of the past in the markets of the West,
just as they have already passed out in the eastern markets. Even
at Clarksville hogshead tobacco auction sales have become almost
nominal in volume, and in Cincinnati for 1909 such public offerings
amounted to only 5,381 hogsheads, as against 70,855 hogsheads so
offered in 1906. In addition, however, to the 5,381 hogsheads sold
publicly in 1909 there were 12,821 disposed of privately.
In Louisville the same tendency is also distinctly shown. The
annual trade reports issued by the Louisville Leaf-Tobacco Exchange
are not published in such form as to make possible a distinction be-
tween the auction and private sales. Weekly reports are issued in
this form, however, and this tendency for an increasing preponder-
ance of private rather than auction sales is distinctly shown in these
reports. It is a matter of common knowledge also that for many
years nearly all the Green River tobacco handled in Louisville has
been sold privately.
LOUISVILLE AND CINCINNATI THE ONLY DISTINCTIVE HOGSHEAD INSPEC-
TION AND AUCTION MARKETS.
To sum up the main features concerning the present position of the
western tobacco markets, we may note that Louisville and Cincinnati
are the only remaining distinctive hogshead inspection and auction
markets.
‘Except the relatively unimportant cigar-leaf breaks, Cincinnati
deals only in Burley !eaf. Its final position as a leaf market, there-
fore, seems to hinge largely upon two main factors, the continuance
of the pooling movement in the Burley district and the permanent
2 ae
o4 TOBACCO MARKETING IN THE UNITED STATES.
success and spread of the loose-leaf tobacco auction system in the
country market centers.
Louisville is the one great central market of the West where all
types of western leaf, Burley, dark-fired, Green River, one-sucker
tobacco, etc., are dealt in. The changed market conditions as they
exist have seemed to accentuate its distinctiveness as the one great
public hogshead market of the West of the clearing-house type.
Figures 8, 9, and 10 show familiar market scenes in Louisville, illus-
trating characteristic stages in conducting the inspection and auction
sale of tobacco in the West.
lig. 8.—-Breaking a hogshead and drawing a sample at a tobacco inspection,
Louisville, Ky.
Neither Louisville nor Cincinnati is situated directly in the im-
portant tobacco-producing territory, and the trade of these centers
is entirely in hogshead tobacco.
In every other western market a-large part of the trade consists
in loose-leaf tobacco purchased directly from farmers, after which
the leaf is rehandled and prized into hogsheads, and a portion of it
may then appear among the hogshead receipts of the same or some
other market for storage, inspection, and sale.
CLARKSVILLE, TENN., THE MOST IMPORTANT DARK-TOBACCO MARKET.
Clarksville, Tenn., continues to be by far the largest distinctive
dark-tobacco market, and in total volume of leaf handled, counting
268
t
PRESENT STATUS OF THE WESTERN MARKETS. 55
none twice, it probably now ranks ahead of Cincinnati and next to
Louisville. More than 30,000,000 pounds of leaf are handled there
annually, a large part of which is received loose. Although its hogs-
head auction-break sales have shrunk to insignificant proportions, it
remains, nevertheless, a very important hogshead storage and in-
spection market. Sales however, are generally conducted privately.
From time immemorial Clarksville has been justly proud of the dis-
tinction, both at home and abroad, of representing the highest
erades of dark leaf produced in the West. This high-grade tobacco,
however, was drawn largely from the adjoining county of Robert-
son, Tenn., and the southern part of Logan County, Ky., particularly
along the course of the Red River and its tributaries. In this sec-
Fie. 9.—An auction-break sale, Louisville, Ky, The sale is based on the official
sample shown on the top of each hogshead of tobacco.
tion is produced the largest percentage of really fine dark leaf to be
found anywhere in the West, such as the fine, rich tobacco suitable
for wrappers for plug, the fine leafy Austrian and Italian cigar-
wrapper grades, and the fat, rich but fine German and English
spinning leaf. |
THE SPRINGFIELD, TENN., MAKKET.
Prior to 1904 Springfield, the county seat and natural trade center
of Robertson County, Tenn., was merely a receiving and rehandling
point, and the tobacco received after being prized was shipped else-
where, principally to Clarksville, for inspection and sale.
In 1904, however, the Planters’ Protective Association established
at Springfield one of its more important inspection and _ selling
268 .
56 TOBACCO MARKETING IN THE UNITED STATES.
agencies, and Springfield to-day has become a really important in-
spection market, ranking probably next to Clarksville in the volume
of its hogshead trade, and because of the fine grade of leaf produced
in the territory which it serves outranks Clarksville and all other
dark-tobacco markets of the West in the average quality of its
offering.
Over 5,000 hogsheads of the 1904 crop and about 14,000 hogsheads
of the 1908 crop were handled at Springfield. Four large storage
warehouses have been constructed there for the accommodation of
the trade.
Fic. 10.—Recoopering hogsheads of tobacco after inspection, sampling, and sale,
Louisville, Ky.
THE PADUCAH, KY., MARKET.
Paducah is an important western dark-tobacco market, standing,
historically at least, as the leading market center for the Padueah
or western district. In the total quantity of leaf handled, however,
it is generally equaled and sometimes surpassed by the Mayfield, Ky.,
market, in the same district but in the adjoining county of Graves,
Paducah, however, remains more of a center for hogshead tobacco
shipped from all points of the district, while Mayfield is more dis-
tinctly a loose-leaf tobacco market and its receipts are more local
in origin. The total receipts of tobacco of all classes at Paducah
amounted in the trade year 1909-10 to about 17,000,000 pounds, of
which about 10,000,000 pounds came in loose and 7,000,000 pounds
in hogsheads.
268
PRESENT STATUS OF THE WESTERN MARKETS. 57
OTHER IMPORTANT MARKET POINTS.
As already noted, Mayfield and Hopkinsville, although handling
u large quantity of tobacco, are now principally loose-leaf tobacco
receiving and rehandling centers rather than important hogshead
markets, as are also Owensboro and Henderson.
Next to Clarksville, but some distance behind in total amount of
tobacco trade, there are six market centers, all handling about the
same gross volume of business. They are Owensboro, Henderson,
Springfield, Hopkinsville, Paducah, and Mayfield, each handling
approximately 15,000,000 to 20,000,000 pounds or more of leaf an-
nually. There are no other western dark-tobacco markets that
handle as much as 10,000,000 pounds of leaf yearly, although there
are many small receiving and handling points doing a strictly local
business, scattered here and there throughout the entire producing
territory. Some of these minor local centers, however, do quite a
large business in the aggregate, amounting annually in many cases
to 5,000,000 pounds or more of leaf.
IMPORTANT RECEIVING POINTS IN THE ONE-SUCKER DISTRICT.
Perhaps special mention should be made of the leading receiving
centers of the one-sucker territory in Kentucky. Glasgow, Bowling
Green, and Scottsville are the principal points, each handling, respec-
tively, about 6,000,000, 4,000,000, and 3,000,000 pounds of tobacco
annually. Practically all the leaf tobacco however, of this one-
sucker district, except such as goes directly into the manufacturer’s
or exporter’s hands, or such as is controlled and sold by the growers’
pooling organization, is sent to Louisville for inspection and sale.
TRADE ORGANIZATION AND MARKET REGULATION IN THE WESTERN
MARKETS.
When the first inspections were established in the western markets
the method followed in nearly all cases was to have the inspectors
appointed by some public agency, as, for example, the city council,
the city or county courts, the mayor of the city, the governor of the
State, or, as in Cincinnati, by the direct vote of the people of the
city. Statutes were enacted for regulating the trade in the interests
of fair dealing between the members of the trade and farmers, par-
ticularly for the purpose of giving the inspection a better standing
abroad than any system of private inspection and regulation would
have done at that time. The laws of Virginia and Maryland natu-
rally served as the basis of the earlier warehouse and inspection
laws of the West.
268
58 TOBACCO MARKETING IN THE UNITED STATES.
In the decade 1850 to 1860 nearly all the larger inspection markets
had begun to feel the need of trade organization, and such organiza-
tions were established in the more important of these markets during
that period.
As in Virginia, it was not many years, particularly in that strained
period of readjustment of the Government in the States of the South
immediately succeeding the close of the Civil War, before the public
appointment of tobacco inspectors became such a mere political plum,
with but little regard to the fitness of the appointees, as to render the
system exceedingly unsatisfactory to the trade both at home and
abroad.
In the early seventies this dissatisfaction with the State inspection
system had reached such a point as to result generally in the taking
over of this function by the organized trade, with sufficient changes
in the existing statutes where it was necessary to render this action
legal. Under this board of trade system of inspection the general
plan was to elect the inspectors by the vote of the individual members
of the entire organized body. This trade organization now became
in effect the guarantor of the integrity of the inspection and sample,
and its inspectors were placed under bond for the faithful perform-
ance of their duties.
This new system of the semipublic nature in turn has sometimes
fallen into disfavor, particularly among growers, who have claimed
they were not’ given sufficient consideration and guarantee of fair
treatment by the board of trade system, and on several occasions, par-
ticularly after periods of low prices, there have been vigorous move-
ments in nearly every interested State to reestablish official State
inspections on a compulsory basis. Thus far, however, the organized
trade has possessed sufficient influence to prevent the enactment of
such measures.
The general plan of organization and the rules under which these
chartered tobacco-trade bodies in the western markets operate are
quite uniform in their essential points. A brief general description
of the Louisville trade organization, or tobacco exchange, as it is
called, perhaps will be sufficient to give a general idea of the way in
which the tobacco trade is conducted in the western markets.
ORGANIZATION OF THE LOUISVILLE TOBACCO TRADE,
Membership in the Louisville Leaf-Tobacco Exchange is obtained
by vote of the members and by purchasing a share of stock in the
exchange and the payment of annual dues.
The members are divided into buyers and warehousemen, the class
of membership being designated on the certificate of stock. An
interesting point in the organization of the tobacco trade of Louis-
268
‘ee Pagid
PRESENT STATUS OF THE WESTERN MARKETS. 59
ville and most other western markets is that on nearly every question
the buying interests and the warehouse interests, that is, the selling
interests, have equal votes. Thus, the executive committee has three
members who are warehousemen and three who are buyers. The
members of this committee are elected, in turn, by a two-thirds vote
of the warehousemen and a two-thirds vote of the buyers. Similarly,
the committee on arbitration and the committee on by-laws consist
of six members each, three of whom are buyers and three warehouse-
men, but in the first-named committee the buyers elect the warehouse
members and the warehousemen elect the buyers on the committee.
In the committee on by-laws each side elects its own members. The
quotations committee, consisting of two buyers and two warehouse-
men, with the secretary of the exchange, is appointed by the president.
On the first Monday in December of each year a joint committee
on inspection is elected, as follows: Each warehouse member is rep-
resented by one member of his firm and a corresponding number of
buyers are elected by the buyers. On the second Monday in Decem-
ber this joint committee elects an inspector to serve for four years
to succeed one of the four inspectors whose term will expire that year.
For any neglect of duty this jomt committee of inspections may
remove an inspector at any time by a majority vote and elect his
successor.
The important committee on reclamations consists of eight mem-
bers, four of whom are buyers and four warehousemen. The ware-
house members of this committee are elected by the warehousemen
and the buyers are elected by the buyers. Two warehousemen and
one buyer constitute a quorum on this committee, and when the buyer
and one warehouseman shall agree in adjudging a case coming before
it on any given day their decision is final. If these two fail to agree,
however, they refer the question to the president of the exchange,
whose decision is final.
Claims on tobacco shipped to points in the United States are not
considered after six months from the date of inspection nor those to
foreign countries after seven months from the date of inspection.
The inspectors and their sureties are jointly hable for such reclama-
tion as the committee may allow. In making a claim the claimant
deposits $1 with the committee. If the claim is allowed, the in-
spectors and owners of the tobacco are assessed $1 in excess of the
amount of damage allowed to be reimbursed to the claimant. To
substantiate reclamations the original sample, properly sealed and
tied, together with a sworn resample of the same hogshead, must
be returned to Louisville. Ifthe hogshead complained of is in Louis-
ville, the inspector must be notified before the tobacco has been taken
from the hogshead and he must promptly examine such hogshead.
268
60 TOBACCO MARKETING IN THE UNITED STATES.
Inspectors have the right to prosecute claims against the sellers of
any tobacco against which damage has been assessed.
In sampling tobaccos no uniform method is prescribed, but the
hogshead is generally broken in at least three nae and a sample
not to exceed 10 pounds in weight is obtained. Usually about 12 or
16 hands are taken. The layers of the sample are tied in the same
order in which the tobacco stands in the hogshead. When a hogshead
of tobacco is damaged in any way, the character of such damage is
marked on the tag in ink. On the tag there also appears in ink the
packer’s name, the number of the hogshead from which the sample
was taken, the date of inspection, and the gross and net weight.
The tag and sample are tied with strong cord and sealed with wax,
using the registered seal of the exchange.
In selling tobacco at auction the hogshead must be inspected on the
day of sale and the sample placed on top of the open bulk of the
hogshead in full view of the buyers. -
The tobacco is weighed before sampling and after sale, and on /
the Louisville market the warehouseman collects from the buyer by
first weight and settles with the seller at 10 pounds less than this
veight. A gain on the part of the warehousemen of over 5 pounds in
sampling, however, is not allowed. The buyer, of course, gets the
sample. .
In selling tobacco at auction the time consumed on each hogshead
must not exceed one minute. The minimum bid on tobacco up to $6
is 5 cents per 100 pounds; up to $10, 10 cents per 100 pounds, except
to round up; and over $10, 25 cents per 100 pounds; and over $20,
50 cents per 100 pounds.
An owner may reject the sale of any hogshead of tobacco by serving
notice on the buyer of such rejection within two hours after the
closing of the sale, but the buyer in turn may proceed to reject an
equal number of hogsheads within two hours after the receipt of
such notice, unless the owner accepts or rejects each hogshead at
the time it is knocked out.
Warehousemen must furnish good storage for the tobacco, the
president of the exchange appointing two members, a buyer and a
warehouseman, who, acting unanimously, have authority to condemn
premises unsuitable for tobacco storage. After a sale the ware-
housemen must recooper each hogshead into first-class shipping
condition,
WAREHOUSE FEES.
The fees collected by the warehousemen are not strictly uniform
in all the markets. The usual charges, however, are either $1.50 or $2
per hogshead and 1 per cent commission to the seller and an outage
fee of $2 per hogshead to the buyer. The Louisville rate to the seller
268
RECEIPTS AT THE IMPORTANT HOGSHEAD-TOBACCO MARKETS. 61
is $1.50 per hogshead and 1 per cent commission. Storage is free to
the shipper for 4 months and to the buyer for 15 days. After
four months the seller is charged 40 cents per month or fraction
thereof for storage, in addition, of course, to such general charges
as insurance, freight, drayage, cooperage, etc. If the sale of a hogs-
head of tobacco is rejected, $1.50 is charged unless the hogshead is
removed, when the rejection fee is $2.50 per hogshead and storage is
charged from the date of receipt.
Reduced selling fees are charged buyers who are members of the
exchange on tobacco purchased from members of the exchange. Thus
buyers may resell tobacco at Louisville within 30 days for $1 per
hogshead or for $1.50 per hogshead after 30 days.
CONSOLIDATION OF WAREHOUSE INTERESTS.
At the present time there are in Louisville 12 warehouses in active
operation and a number of others once active but now used only for
storage purposes. Of these 12 actively operated 8 are under joint
ownership, though operated separately under the control of the
Louisville Warehouse Co. One of these 12 is not even a member of
the exchange.
In Cincinnati hkewise, although there are five houses in actual
operation, four of them are consolidated under the ownership of the
Cincinnati Warehouse Co. In 1890, in fact, a movement was re-
ported looking toward the combination of both Louisville and Cin-
cinnati under single ownership and control.
PRIVATE WAREHOUSE INSPECTIONS,
In the last few years, owing perhaps to the general disturbance
within the regular channels due to changing conditions, the trade
bodies in some instances have been allowed to fall somewhat into
decay, and except as a matter of custom exert very little influence
over the trade.
In some of the markets the inspectors, instead of being appointed
and controlled by the trade organizations, are simply employees of
the warehouse company, and the warehouse, acting without outside
regulation by the organized trade, adjusts its own claims for recla-
mation.
_ In Tennessee the law specifically provides for the appointment of
warehousemen as inspectors under bond to the State.
SUMMARY OF THE RECEIPTS AT THE IMPORTANT HOGSHEAD-
TOBACCO MARKETS FOR 10 YEARS.
In the case of the Mayfield, Paducah, Hopkinsville, and Clarks-
ville markets the figures in Table XI of the receipts of hogshead
tobacco are not to be taken as indicating anything like their total
268
62 TOBACCO MARKETING IN THE UNITED STATES.
a
general leaf and brokerage trade. A considerable proportion of the _
trade of these markets, and the larger portion in some cases, is in
loose tobacco bought by manufacturers or exporters direct from the |
farmers and does not appear in the regular market receipts of any
market, as already explained. Some of the figures in Table XI were
obtained from the compilation made by the Western Tobacco Journal.
TABLE XI.—Receipts of leaf tobacco in the principal hogshead markets of the
United States from 1900 to 1909, inclusive.
Market. | 1s | 1901 | 1902 | 1903 | 1904 | 1905 | 1906 | 1907
Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs-
heads. | heads. | heads. | heads. | heads. | heads. | heads. | heads.
Bomisville..: 22 2222-2 106, 827} 123, 279] 124,213] 80,051] 84, 104} 100,335] 105,973) 107,525
Cmeinivsil: . ...2- 2: 56,070} 60,318] 51,638) 52,093] 21,022) 45,419) 55,380) 37,317
Clarksville: _.... 27-222. 20,501} 22,322) 21,791) 20,843) 21,220) 22,980) 9,847) 11,533
Hopkinsville. ....... 14,165} 12,465) 11,975) 11,350} 14,930) 9,715) 5,450) 4,655
Sprimefield ....-:.~..--|s% <.<200)osaeee eae eee eeeee eee ee 4,500} 7, 8, 700
Paaurale. 222): 1c. 9,987; 7,273} 8,697) 11,000} 8,690] 5,996) 5,381; 6,311
Mavhdld. - 22222055 12,518} 9,780) 10,594; 7,995) 10,770) 8,039) 5,481) 4,569
Richmond. ....-.--: 27,663) 21,522) 20,096) 21,150) 17,487} 23,330) 20,404) 19, 636
Baltimore.:...-..2-. 38,023} 35,881] 39,480} 40,051] 40,734) 38,563) 37,055, 25,594
Total:...s2ncs 285, 754| 292, 840} 288, 484) 244, 533] 218,957| 258, 877| 252,471, 225, 840) 216, 286) 176, 785
TABLE XII.—Jnventory of the stock of tobacco on hand on Jan. 1 in the prin-
cipal hogshead-tobacco markets for each of the 10 years from Jan. 1, 1901,
to Jan. 1, 1910, inclusive.
Market. 1901 1902 1903 1904 1905 1905 1907 1908 1909 1910
Louisville. 22-2 13,031 | 15,627 | 12,266 | 11,796 | 18,001 | 16,215 | 17,906 | 27,078 | 23,978 15,737
OiMneinnat 223 2 9,391 | 12,287 | 10,084 8,781 6,238 9,080 8,955 | 11,278 | 14,234 4,312
Clarksville__......| 2,662 581 } 1,583 | 3,348] 1,049 | 3,560 579 329 | 3,780 740
Hopkinsville_....| 3,124 808 381 | 2,485] 1,392 | 1,671 374 210 | 2,000 109
Springfield 1__.=..|...2..5.|22c-225-|---- elec eZ) oe ee ne ee
PAGUCHOE~—.=s2cs. 824 PON Teil || 2Aao 976 517 251 376 | 6,500 225
Mayfeld-.....-<...| “1,587 222 | 1,624 825 291 727 144 | ee 1,400 423
Richmond._.._...-- 12,020 | 9,839 | 9,326 | 9,326 |- 8,230 | 10,659 | 13,065 | 12,506 | 13,799 | 17,181
‘Balimores - 30 2_* | 6,425 5,521 4,989 / 7,159 |\. 5,240 4,925 6,111 3,300 3,891 3,065
Total... .:.2| 56,074 | LP, O5G | 41,414 | 45,819 | 41,417 | 47,354 | 47,385 | 55,077 | 69,582 | 41,783
| |
1 No record.
The total stocks on hand on these principal markets have averaged,
according to the figures in Table XII, not far from 50,000 hogs-
heads of tobacco on January 1 of each of the 10 years:specified. This,
of course, does not include that stored privately in dealers’ hands
or stored privately in the hands of manufacturers. The latter figure
would reach a very large total, as it is customary for manufacturers
to carry from one to two years’ stocks, and many of the larger firms
own private storage plants.
These stocks held privately by manufacturers amount probably to
at least 400,000,000 pounds at any given time, and often, no doubt, —
to very much more than this, against something like 60,000,000 to —
80,000,000 pounds generally held in the public warehouses of the —
regular markets.
268
a pees
NET Sorted Ba eee y
ESTIMATING THE AVERAGE ANNUAL PRODUCTION. 63
The Louisville tobacco trade is well organized, with a salaried
secretary, and publishes very complete annual statistics of the trans-
actions on that market, including the classification of sales. Since
Louisville is the largest and most important hogshead-tobacco market
in the country, the Louisville leaf-trade statistics are given in Table
XIII for each of the past 10 years. They throw considerable light
on certain trade features not before mentioned, such as differentiation
between offerings, sales, rejections, reinspections, etc.
TABLE XIII.—Statistics of the Louisville market for 10 years, from the report
of the secretary of the Louisville Tobacco Exchange.
Item. 1900 1901 | 1902 1903 1904 1905 1906 | 1907 1908 1909
Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs- | Hogs-
heads. | heads. | heads. | heads. | heads. | heads. | heads. | heads. | heads. | heads.
Offerings............ 145, 339] 156,788] 164, 696] 104, 241] 107,007] 140, 112| 143, 784] 125, 673| 118,612] 95,967
Rejeciions....... ...| 29,123] 28,851] 32°159| 17,989] 12,967] 18,831] 14,986] 13,559] 9,341] 8,77
Actual sales......... 116, 216] 127, 937] 132,537| 86,252] 94,040] 121, 281] 128,798] 112,114] 109,271] 87,197
Original inspection..| 105,046] 130, 693| 112,966] 87,656] 82,875] 96,200] 118,014] 98,290) 71,289] 66, 494
Heceipts............ 106, 827| 123, 279| 124,213] 80,051] 84,104] 100,335] 105,973] 107,525] 97,099] 75,190
Total stock Dec. 31..} 13,031] 15,627| 12,266] 11,796] 18,001] 16,215] 17,906] 27,078] 23,978] 15,737
Unsoldstock Dec.31.| 9,612} 12,733] 9,920] 10,622] 14,835] 10,717] 12.580] 22,306] 14,898] 9,446
Unsold Burley..... . 3,803} 9,160} 7,832} 1,869} 1,158) 2,096] 3,688) 9,767) 12,676} 2,233
Warsold’dark._...... 5,809} 3,573] 2,063) 2,353} 6,934) 2,296] 1,870) 1,243) . 1,915) 4,484
Wusold Green River.|........\.....-..- 25| 6,400) 6,743] 6,325) 7,022) 11,296 307| 2,729
Classification of sales:
: Ln 99, 969} 116,094] 120,733} 70,279} 43,554) 86,512} 90,345) 71,484) 77,547] 51,752
likh) aa 45,370} 40,694) 43,963) 33,962} 63,453] 53,600) 53,439) 54,189) 41,065} 44,215
Rejections |
elt) 21,375} 21,984) 24,132); 11,726} 4,963] 13,326) 12,075} 10,344) 5,481) 3,609
ih 7, 748) 6,867) (8,027) 6,263) 8,004) 5,505; 2,911)° 3,215) 3,860} 5,161
There is a small error in the foregoing figures, since they fail to
show the full volume of the tobacco-leaf trade of Louisville. Of the
12 warehouses in operation in the city in recent years, one is not a
member of the exchange and its transactions are not included in the
report.
METHOD OCF ESTIMATING THE AVERAGE ANNUAL PRODUCTION
OF TOBACCO IN THE UNITED STATES BASED ON STATISTICS OF
THE TREASURY DEPARTMENT.
On their face, the fully itemized records of the offices of Internal
Revenue and Customs of the Treasury Department, specifying with
substantial accuracy the quantity of raw tobacco imported and ex-
ported and the quantity that is consumed in manufacture, should
furnish the data for estimating the actual farm production of tobacco
in the United States on a very reliable basis.
It would appear as though a statement of the quantity of raw to-
bacco used in manufacture plus the exports and less the imports of
raw tobacco for a year or for a series of years would be equal to the
farm production of tobacco in the country. |
There are, however, a number of important sources of error to
which production figures derived in this way would be subject, in so
268
64 TOBACCO MARKETING IN THE UNITED STATES.
far as they purported to serve as a check on the actual weights of
barn-cured tobacco reported by farmers.
The factors of importance producing this error are the following:
(1) The shrinkage to which leaf tobacco is subject while it is in
the hands of dealers or manufacturers because of the redrying to put
it in good keeping order and while it remains in storage. This shrink-
age in weight from the time the unstemmed leaf tobacco leaves the
farmers’ hands till it enters the manufactory or is cleared for export
ranges from 5 to 20 per cent, according to the type or tobacco and
its moisture content at the time of purchase from the farmer.
(2) In the case of leaf tobacco which is stemmed before it is
weighed in the manufacturing plant or is declared for export, there
is an additional loss due to the weight of stem removed. The loss in
weight to which stemmed tobacco has been subjected, including the
drying as well as the stemming, is estimated to be from 28 to 35 per
cent,
(3) There is also considerable tobacco not recorded, which is con-
sumed in its natural state locally by producers, dealers, farm hands,
laborers, and others, destroyed by fire, etc. This source of error
all told is believed to amount to about 1 per cent of the total produc-
tion, and, while small in respect to the whole, it is of considerable
importance in the aggregate,
The total of these three sources of error makes so large an error as
to render an uncorrected compilation of the statistics of the Treasury
Department of but little service in estimating the farm production.
The question which at once presents itself is to what extent and with
what degree of accuracy can corrections be introduced into the re-
corded figures to make them of such service. As a matter of fact,
this seems comparatively easy to do with approximate accuracy and
to such a degree, the writer believes, as to render them the most accu-
‘ate basis for estimating the average annual crop of tobacco produced
in the United States for any period of years, preieee not less than
three.
The shrinkage of leaf tobacco in redrying or stemming and in
storage after it leaves the farmers’ hands is a matter of regular esti-
mate by leaf dealers and manufacturers who make direct purchases
of leaf, and these estimates are accurate to a very small percentage of
error where a sufficient number of pounds for each of the different
types produced in the country are involved. The actual figures, of
course, vary somewhat for the different types of tobacco, but a general
average correction applicable to all types would doubtless give an
average result of sufficient accuracy for general purposes.
The most important missing item in the figures of the Treasury
Department is the fact that exports of stemmed and unstemmed Jeaf
are not reported separately, but are classified together under one
268
f
<
=
5
_ ESTIMATING THE AVERAGE ANNUAL PRODUCTION. 65
head. It so happens, however, that a very large proportion of the
stemmed tobacco exported from this country goes to the United
Kingdom, and the trade reports of the United Kingdom separate the
stemmed leaf from the unstemmed, so that an approximately correct
statement is available as to the quantity of stemmed leaf tobacco
which is included in our export figures.
Production estimates based on compilation of these figures of the
Treasury Department, with proper corrections, of course, would not
give much of an idea of the size of the crop for any particular year.
The exports of leaf tobacco are generally made during the year suc-
ceeding its production, but stocks of tobacco for domestic manufac-
ture are generally held from one totwo years in order that the tobacco
may improve and mellow with age before it is manufactured. To be
of value the compilation, as suggested, must represent the average
of figures covering three to five years, and as such they should give
an accurate average estimate of the rate of production for a period
of three to four years, dating back one year from those figures from
which the compilations were made.
Such a compilation, based on the statistics of the Treasury Depart-
ment for the three years 1907 to 1909, inclusive, is presented below.
In considering the quantity of tobacco consumed in producing the
so-called manufactured forms of tobacco, namely, plug, twist, fine-
cut smoking tobacco, snuff, and cigarettes, and the quantity of
exported leaf, we have to deal almost entirely with the so-called
export and manufacturing types. We will assume that, as a general
average, these types shrink in redrying and handling about 10 per
cent if unstemmed and about 30 per cent if stemmed. In the cigar
types the shrinkage is probably somewhat greater and will be cal-
culated at 15 per cent for the unstemmed leaf. The correction to an
approximate equivalent of unstemmed but dry-leaf is made by the
‘office of Internal Revenue in the case of the stemmed leaf used in the
manufacture of cigars and cigarettes, so this factor does not need to
be considered here.
We will first consider the leaf used in producing manufactured
tobacco and cigarettes and for export. In the 3-year period, 1907 to
1909, inclusive, there were consumed in manufactured tobacco and
cigarettes 283,550,157 pounds of stemmed leaf, 631,736,223 pounds of
unstemmed leaf, and 99,785,876 pounds of scrap. The exports of
domestic leaf for the same period amounted to 957,080,408 pounds.
The import statistics of Great Britain show that at least 147,145,070
pounds of this latter total was stemmed leaf. We also export some
little stemmed leaf to countries other than Great Britain. It would
be well within the facts, therefore, to assume that not less than 150,-
000,000 pounds of stemmed leaf were exported during the 3-year
period. Combining this with the stemmed leaf reported by the office
65602°—Bull. 268—13———5 7
66 TOBACCO MARKETING IN THE UNITED STATES.
of Internal Revenue, we have a total of 433,550,157 pounds of
stemmed leaf of the export and manufacturing type to which the 30
per cent correction should be applied. We find this to have been
equivalent to 619,357,367 pounds of unstemmed leaf, based on farm-
ers’ weights.
The scrap tobacco reported by the office of Internal Revenue con-
sisted largely of cigar clippings and other material, much of which
perhaps had already been corrected for loss of stem. For our pur-
poses, therefore, it may be classed with the unstemmed leaf, to be
corrected for loss simply for shrinkage from drying and age.
Making this correction of 10 per cent, we find that this total of
1,689,328,514 pounds of unstemmed leaf of the export and manufac-
turing types used in domestic manufacture and exported to have
been equivalent to 1,877,031,682 pounds of tobacco at farm weights.
The farm weight of the combined total, stemmed and unstemmed,
manufactured, and exported, was, therefore, during the 3-year period
about 2,496,389,049 pounds.
The quantity of unstemmed leaf consumed in the manufacture of +
large and small cigars during the three years amounted to 414,636,193 a
pounds. Allowing for a shrinkage of 15 per cent this was equivalent
to 490,160,227 pounds, farmers’ weight.
The export factories under the supervision of the Customs Service
consumed 25,462,093 pounds of leaf, not included in the foregoing
estimates. It is uncertain how much of this was stemmed weight,
but as the quantity involved is small, it is not very material. It is,
however, subject to the 10 per cent correction for shrinkage in
drying. Making this correction we find this to have been equivalent
to 28,291,214 pounds.
Combining these three totals we have a grand total of leaf con-
sumed in manufacture and export, reduced to farmers’ weights of
unstemmed leaf, as follows:
Export and manufacturing types exported and reported Pounds.
by office of Internal Revenue___-_------------------~ 2, 496, 389,,049
Consumed in clwars. 225. 5- > > Ae ee > 490, 160, 227
Consumed in export factories reported by the customs
office uo. jue ck eee ee ith shige SS She aS RE 28, 291, 214
Total for three years....__/-i__.- Saba Sa
From this total must now be deducted the quantity of leaf imported
for consumption during the 3-year period, including that from
Porto Rico.t| This imported leaf, however, has been included in the
tobacco to which the foregoing corrections to a farm- weight basis
were applied. In subtracting the imports as recorded by the Cus-
‘The deduction of imported leaf from Porto Rico was for fiseal-year periods; all ota
figures were for calendar-year periods. ;
268
7
}
3
ESTIMATING THE AVERAGE ANNUAL PRODUCTION. 67
toms Service, therefore, the figures must also first be raised to a
farm-weight basis. About four-fifths of this imported leaf was of
the cigar type and the remainder of the Turkish type. A compro-
mise figure of, say, 14 per cent instead of either 10 or 15 per cent,
as used in correcting the manufacturing types and cigar types, would
give only a small error either way.
The imports for consumption, including those from Porto Rico.
during the 3-year period amounted to 119,062,219 pounds. Making
the correction for the estimated shrinkage of 14 per cent, we find the
computed farm weight of this imported leaf to be 138,569,051 pounds.
Subtracting this quantity from the total consumed and exported,
as shown, we have a total of 2,876,271,489 pounds.
Adding to this the 1 per cent estimated to have been consumed on
the farm, destroyed by fire, and otherwise unaccounted for, we have
a grand total, corrected to an approximate unstemmed farm-weight
basis, of 2,905,324,686 pounds of domestic tobacco consumed in the
United States and exported during the three calendar years 1907,
1908, and 1909. Dividing this total by 3, we have 968,441,562 pounds
as the indicated average annual production of leaf tobacco in farm
weights in the continental United States within the approximate
period, say from 1906 to 1908.
These computations have been submitted mostly as an illustration
of the method by which the official Treasury records might be utilized
in estimating the average annual production of tobacco in the United
States on a farm-weight basis. Possibly there are still some im-
portant sources of error in the method that have failed to receive
proper consideration. The percentage basis upon which the correc-
tions were based, 10 per cent for unstemmed leaf of the export and
manufacturing types, 15 per cent for the cigar types, and 30 per cent
for the stemmed leaf of the export and manufacturing types, are, as
stated, assumed figures of a somewhat arbitrary character.
268
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Issued April 24, 1913.
U. S. DEPARTMENT OF AGRICULTURE.
BUREAU OF PLANT INDUSTRY—BULLETIN NO. 269.
B. T. GALLOWAY, Chief of Bureau. \
EXPERIMENTS IN WHEAT BREEDING:
EXPERIMENTAL ERROR IN THE NURSERY AND
VARIATION IN NITROGEN AND YIELD.
BY
E. G. MONTGOMERY,
Experimental Agronomist of the Nebraska Agricultural Experiment Station
: and Collaborator of the Office of Cereal Investigations,
Bureau of Plant Industry.
WASHINGTON:
GOVERNMENT PRINTING OFFICE.
1913.
BUREAU OF PLANT INDUSTRY.
Chief of Bureau, BEVERLY T. GALLOWAY.
Assistant Chief of Bureau, WILLIAM A. TAYLOR.
Editor, J. E. ROCKWELL.
Chief Clerk, JAMES E. JONEs.
CEREAL INVESTIGATIONS.
SCIENTIFIC STAFF.
Carleton R. Ball, Acting Cerealist in Charge. P
Charles E. Chambliss, H. B. Derr, H. V. Harlan, and C. W. Warburton, A gronomists.
E. L. Adams, Assistant A gronomist.
Clyde E. Leighty, Expert. :
Cecil Salmon, Physiologist.
John F. Ross, Farm Superintendent.
A. A. Potter, Assistant Pathologist.
L. C. Aicher, P. V. Cardon, Manley Champlin, J. A. Clark, N. C. Donaldson, J. Mitchell Jenkins, E. M.
Johnston, Jenkin W. Jones, F. A. Kiene, jr., Clyde McKee, J. D. Morrison, B. E. Rothgeb, T. R. Stanton,
and L. Wermelskirchen, Scientific A ssistants.
F. R. Babcock, Assistant.
L. R. Breithaupt, L. C. Burnett, and H. H. Love, Agents.
D.E. Stephens, Executive Assistant.
269
2
LETTER OF TRANSMITTAL.
U.S. DEPARTMENT OF AGRICULTURE,
BuREAU OF PLANT INDUSTRY,
OFFICE OF THE CHIEF,
WasuHineton, D. C., October 16, 1912.
Sir: I have the honor to transmit herewith and to recommend for
publication as Bulletin No. 269 of the series of this Bureau the
accompanying paper entitled ‘‘Experiments in Wheat Breeding:
Experimental Error in the Nursery and Variation in Nitrogen and
Yield,” by Prof. E.G. Montgomery. This paper contains the results
of special experiments in wheat breeding conducted by the Nebraska
Agricultural Experiment Station in cooperation with the Office of
Cereal Investigations of this Bureau, during the years 1905 to 1910,
inclusive. In part, the work is a continuation of that recorded in
Bureau of Plant Industry Bulletin No. 78, by Dr. T. L. Lyon, under
whose direction the experiments were conducted from 1902 to 1906,
inclusive. From 1907 to 1910, inclusive, the work was under the
charge of Prof. Montgomery, experimental agronomist of the Ne-
braska experiment station, who has since become professor of farm
_ crops in the College of Agriculture at Cornell University.
The paper is concerned chiefly with the nature and extent of
experimental error in the wheat nursery in connection with breeding
experiments in the inheritance of nitrogen content and yield in wheat
plants. The standardization of agronomic experiments has been
receiving much attention in recent years and is regarded as of fun-
damental importance in agronomic research. The results contained
in this paper are presented as a contribution to this subject as well as
to the improvement of wheat.
Respectfully, B. T. GaLtoway,
Chief of Bureau.
Hon. Jamrs WILSON,
Secretary of Agriculture.
269
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CONTENTS.
I. Experimental error in the nursery and variation in nitrogen content......-
PP AMIIRC MINS AAS ESS hate UGE oR Sa ih fois es atk he p die’ > Sax cine Melee sR
Relation of yield of grain to nitrogen content................---------
heauring the experimentat-error: st . 125... oe a oe eel cee
ep MCalign OL pita PIA Wae toy. 33 2: 4). o 3 oe's ab tue ela 2 wine
ep ICdeION OF 2-10 OWS Au cise b- o oes cnn bed cal ke eee
Pr emlcaMOmrOL LGAdob FOWHe si. Oh. Vasc AA Loos. f cdeuw nelle ee OS
Peano PaYit biiGie RON. een ays oe ek 6.26 2 e's Soins cota. bei
increasing the. sizeof, plat... 05 -2.-2-2..-. Joo ae eiucas Sicyeiagine ee
fee nd Of experimental errar.o. 42 20.7. 422. 2d ons pe eh oes een oot
MRR 0! Caran UREA a, hone UH ORD. oe aaa pa. aie Se eS ee
II. Experimental error in the nursery and variation in yield................--
MIE IC RUE Sea ia, 2 hoe hs NS © a'h! che’ ON Ys olen 2 ome oe Qe
Seamiaiton. m yield from check Tows.-...:. 2-22 4-.206-1-he6- epee: tS
Maranomin yield from repeated rows-..". 5.2.2... 62.- 420s. - s<-- se tbe
amaiion im yield from small blocks: ....:........-...1....02. Yad aoe
Effect of repetition in reducing error..........-.- Nes eI c's apts eeaiaee See
Poel a Oh RG OL DING LOVATIALION..\.. 2.22405 --a5<080 2-5-2 amen eee
Constancy of variation on,the same plats.............-.-.......-
Wanation in yields irom centgener plats... 22-2 22.-.5-. i. 5+--.0500 02
Alternating check rows as a means of obtaining comparative yields. . .
feet OF MINCKOSAINIS TENGEN OT TOW 04+... 6t cote Pe nae eae es wn dee Se
maomence.of Pate Of planting om Yield. ...<4. 22... 2.2465. 00----00-seneue
Effect of competition between adjacent rows.......-.--.----...-------
Variation in pure strains and relation of data in centgener nursery
0 OS VN EOP SE ce So a
Pere PITS SURAN oe oe ee. as. kn on lows aaa so et
Perret npe OF MAEOMOM 82h erste tase bn hk 4 Sole ase da ocageln =e wee
PUPOMOMLOL CUPGIW 2) as, 2) 22 Nuts oan whet ee vis Sa he chat
(DESO. TS a Ra GS SB Ts ee Se ee We Eee eo
Rio Ot CONES ede oe pane wee ia en tee eek de eee Ok ee
mize or kermely 2. oi -2. 525% Nitta chad eign eee 2 ahs Bes ay i Nee Nags 5h
AST VS 1191) CRS FES SAR Laie 5A aS Paes anne me, eee
PeRmePeMCIUR CLMEPMETDS M7 fe tytn . Ua cate tila g hs Wh aS ae 5 ne
Comparison of rows, centgeners, blocks, and field plats...............-
Cost of planting and harvesting centgeners, rows, and blocks. ........
RMR OOO N ete Toe. Ula en tees Se ON wie wg noo sa wane oem oe
Me IN ok ae oS as Ge wane tia Pe owt se nh anime a oe
Piet precautions AGalNst Crror i ss... oc. os oh be eke eee eee eens:
Accidental injury to plats................. SEIS SE OR SRM ee
apa RCE ENE Ps een. le Sag ets" Se a Sam nw ne tae ws SO
en are eet he ee er fhe) hee 2 hee es
269
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TABLE I.
Il.
IE.
XI.
XII.
mT.
XIV.
XV.
XVI.
».O' 85
XVIII.
XIX.
b..@
XXI.
XXII.
TABLAS
Results of a study of transmission of nitrogen content in wheat ker-
nels in 1902 and: 1903... .. 2s. soso. el eee
Nitrogen content of 57 wheat plants in 1903 and of their progeny in
1904 and 1908, ..0..2...2 52.0... soe bee ee
Data from six wheat plants, showing irregular variation in yield
and in nitrogen content of grain. --o.:5.- 02.2.0 see
. Nitrogen content and yield of grain from families Nos. 42 and 48,
from 1902 to 1905, inclusivé..2.2..2...0..1. 2 te
. Nitrogen content of 29 centgeners and corresponding rows from
family No. 831 in: 1908.2... .4...2500..5.2002_0 2) er
. Nitrogen content and yield of grain from 180 wheat plants, arranged
in inverse order of percentage of nitrogen, in groups of 10........
. Nitrogen content of 90 plants of Turkey wheat from 1 centgener
and of 840 plants variously combined into groups to show devia-
tion from meéan......-..s.e.0.. .2tc ALe eae es
. Nitrogen content of 110 2-foot sections of drill row of Turkey wheat. .
. Nitrogen content of 100 16-foot rows of Turkey wheat............--
. Summary showing degree of error due to variation in environment,
according to several methods of comparison.............---.-----
Nitrogen content of Turkey wheat grown in 224 block plats in 1909
and JOLO. 020s cect een bt eec wack ae tep ecw ee ne on
Yield of 47 14-foot check plats of Turkey wheat in 1909...........--
Yield of thrashed grain from 100 rows of Kherson oats...........-.--
Yield of grain from 500 16-foot rows of Turkey wheat, systematically
repeated in various ways to show experimental error............--
Yield of Turkey wheat grown in 224 block plats in 1909 and 1910...
Yield, in grams, of Turkey wheat grown during the season of 1910 in
500 rows, each 16 feet in length?..... 3. <..2..... 0. So
Summary showing coefficients of variability under various systems
of arranging block plats and row plats.....................---.---
Results of rate-of-seeding test on 100 16-foot rows of Red Rustproof
Results of rate-of-seeding test on 60 block plats of Kherson oats... . .
Relations of certain characters of 24 strains of Turkey wheat grown
in nursery and in field and tested during 4-year periods ........
Yields of grain from 11 varieties of oats grown in field plats, cent-
geners, rows, and blocks. . .<: «0.5... .<.25 200s.» ss» ae ee
Comparative number of plats of different types that can be planted
or harvested in 10 houra.: . af. an. sc csncc eas ec y= 3 55 enn
269
6
Page.
10
*
, ©. 7. = bo
—_—
*,.
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A
ILLUSTRATIONS.
PLATES.
Pirate [. Fig. 1.—Head-to-row nursery, in which 25 grains from a single head
Fig.
are planted in a row 20 inches long. Fig. 2.—Row-plat nursery,
in which the rows are 16 feet in length with a 4-foot alley between
beds, thus making the beds 20 feet in width. . y
II. Fig. 1.—Increase plats of one-thirtieth acre Seek. Fi ig 2, tneeae
Piaes harvested anu ready to thrash... 2)... 25. el ade
III. Fig. 1.—Type of road grader or drag used in grading a nursery into
beds. Fig. 2.—Grains of Turkey wheat, showing variation in
NDC oat Re her Pa IS ate eos il onde ain oe ns 2s, sPL Oe « & Sec, Oe a
IV. Fig. 1.—Representative kernels from four strains of Turkey wheat,
selected to show variation in appearance. Fig. 2.—Representa-
tive kernels from four strains of Turkey wheat, selected from a
series of 80 strains to show variation in quality. ..........----.---
TEXT FIGURES.
1. Diagram showing the transmission of nitrogen content in 57 wheat
mints of 1903 fo progeny in 1904 and 1905.......-:-./.-..---..2-
2. Centgener nursery, Nebraska Agricultural Experiment Station........-
3. Wheat centgener just after growth has started in spring, showing about
ager cent of the plants winterkilled:)........-...-.$...5....)---
4. Wheat centgener of 100 plants, showing variations in yield of grain
peur Tigi neon WY PUN oo ee 5-68 5 ine es oo ba iwc co a7 an Soe cease oe
5. Diagram of a portion of the wheat nursery in 1907, Lig variations
in nitrogen content in centgeners and families. . : :
6. Diagram of 10 centgeners and 10 oe ee rows, oe ing varia-
tions in nitrogen content for individual plants, for each centgener,
for each corresponding row, and also for their parents. .
7. Diagram of plat of Turkey wheat containing 224 blocks, ‘ghowing thé
location of each block and variations in the percentage of nitrogen in
Tee RS Soni SEE De les Bal ar esters Leen Uae Apr ae
8. Diagrams of plats of Turkey wheat, showing the arrangement of 224
blocks when combined in groups of adjacent blocks, with average
minoccn- content Jor Gach sroup.-5 05 -s[)- 2s sos ose ee tae
9. Diagram showing the method of selection for nitrogen content when
imeenmermenian error id KNOWN: 2's.) 5.0.4 gs oo 8-50 oe eee
10. Diagram of plats of Turkey wheat, showing the arrangement of 224
blocks and the yield of grain for each block ..................---.-
11. Diagrams of plats of Turkey wheat, showing the arrangement of 224
-blocks when combined in groups of adjacent blocks, with the aver-
age yield for each group..-.........-...-
269
Page.
42
42
52
52
10
12
13
14
15
18
22
38
12.
13.
14.
15.
16.
Ad,
18.
19.
20.
21.
22.
Diagrams showing Turkey wheat grown in 224 blocks, combined in
four groups of 56 adjacent blocks to show variations in yield and
nitrogen content......
Field plats of pure strains and check plats of original seed of Turkey
wheat, 1910....-2.s..-
ILLUSTRATIONS.
Wheat nursery plats, showing variations in winterkilling........-
Field plats, showing variations in winterkilling between two nlite
strains of Turkey wheat....2-. 2.2253. 00 5.2055 -50h >see se
Increase rows of Turkey wheat, showing variations in the time of head-
ing in different strains... 2. - 22+ on ost e tye ob e- oe
Field plats of Turkey wheat, showing variations in stiffness of straw
in two strains ........
Cereal laboratory, showing the method of taking notes on quality... ..
Block nursery, showing blocks 4.2 by 16 feet im size. ..............--
Five-row nursery drill used for planting row plats and block plats.....
Row plats at harvest time.......2..00.5 S-0s0ade soe s05, be oe
Diagram showing the method of selection for yield when the experi-
mental error is known
269
B. P. I.—785.
EXPERIMENTS IN WHEAT BREEDING: EXPERI-
MENTAL ERROR IN THE NURSERY AND VARIA-
TION IN NITROGEN AND YIELD.
I—EXPERIMENTAL ERROR IN THE NURSERY AND VARIATION
IN NITROGEN CONTENT.
INTRODUCTION.
The investigation of the variation of plants of winter wheat in
relative nitrogen content when grown under field or nursery condi-
tions was begun by Dr. T. L. Lyon, formerly agronomist of the
Nebraska Agricultural Experiment Station, in collaboration with the
Bureau of Plant Industry of the United States Department of Agri-
culture. His results were published as a bulletin of that bureau.t
Since 1907 the investigation has been continued by the writer and
his assistants.’
One of the striking features of the data obtained by Dr. Lyon was
the variation in nitrogen content of the kernels from different plants
of wheat grown under apparently similar conditions. For example,
800 spikes of Turkey wheat were selected and half of each spike
analyzed for proteid nitrogen, the lowest having only 1.12 per cent
while the highest contained 4.95 per cent.
In 1903, 288 plants which were the progeny from 119 of the spikes
analyzing above 3 per cent proteid nitrogen in 1902 were analyzed
and found to vary from 1.20 per cent to 5.85 per cent in nitrogen
content. In most of the families only a single plant was selected
for analysis, but in the remainder two to six plants were selected.
Even where all the plants were grown from a single parent the varia-
tion was quite as great.
1 Lyon, T. L. Improving the Quality of Wheat. Bulletin 78, Bureau of Plant Industry. 1905.
2 The writer wishes to acknowledge with thanks the assistance of a number of men who have contrib-
uted to the production of these data. Dr. T. L. Lyon, now of Cornell University, planted the first wheat
nursery in 1902 and conducted the work until 1906, being assisted by Prof. Alvin Keyser, now of the Colo-
rado Agricultural Experiment Station. They left an excellent set of records, from which data previous
to 1906 have been prepared (Table II). Mr. L. L. Zook, now of the Bureau of Plant Industry, assisted
with the work in 1907 and 1908, as did Mr. Erwin Hopt in 1908 and 1909. Prof. T. A. Kiesselbach had
charge of the records during the seasons of 1909 and 1910 and has prepared much of the tabular matter for
publication. The chemical analysis has been under the direction of Dr. F. J. Alway, who devised a rapid
method especially for this work.
69826°—suL. 269—13———2
10
EXPERIMENTS IN WHEAT BREEDING.
Dr. Lyon noted this variation, as follows:
For instance, the plants numbered 21205 to 21212, all of which come from the same
parent, vary from 2.16 to 5.23 per cent in proteid ‘nitrogen content, while plants
69805 and 69806 vary from 5.82 to 1.66 per cent in this constituent.?
In addition to the 119 ‘‘highs”’ preserved in 1903, progeny were
analyzed also from 20 ‘‘mediums” and ‘‘lows.” When these data
were summarized it seemed that there had been some tendency to
transmit the character, as shown in Table I.
Tasie I.—Results of a study of transmission of nitrogen content in wheat kernels in
1902 and 1903.”
1902 1903
Range in percentage of proteid nitrogen. Number : Number F
of analy Phat S. of analy- oo
ses aver- |") Grnois, | eS aver- | neon me
aged. . | ged. erneis.
| | Per cent. Per cent.
DO S225 2 ek 3 oo eek eke a ee 20 z 70 2.59
BWV: oon 5 52 u nse he hep oe oe Ce en ee 119 | 3. 39 288 2.92
Ww
PEP CENT OF NITROGEN
N
Fig. 1.—Diagram showing the transmission of
nitrogen content in 57 wheat plants of 1903 to
progeny in 1904 and 1905. The vertical lines
_represent successive years. The horizontal
lines represent the percentage of nitrogen
found, and the figures in parentheses show the
number of plants in each group analyzed for
nitrogen content in 1903.
269
Summaries of the results obtained
in 1904 and 1905 show very little
tendency to transmit this charac-
ter.
In 1906, after four years of selec-
tion of extremely high fluctuates,
and later, after two more years of
selection, by taking a composite
sample of all the progeny of a plant
it was found that no gain had been
made in the nitrogen content of the
crop.
In Table II is a summary of data
obtained in the years 1903 to 1905
from 57 of the original plants. Fig-
ure 1 is a graphic: presentation of
the same data, in which the hori-
zontal lines represent the percent-
age of nitrogen, the vertical lines
represent successive years, and the
figures in parentheses show the num-
bers of plants in each of the seven
groups analyzed for nitrogen content
in 1903.
1 Lyon, T. L. Op. cit., p. 99.
2 Lyon, T. L. Op. cit., table 26, p. 98.
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 11
TaBLE II.—Nitrogen content of 57 wheat plants in 1903 and of their progeny in 1904
and 1905, arranged in groups according to the percentage of nitrogen.
Record of 57 orm harvested in Record of progeny plants in 1904. | Record of progeny plants in 1905.
Kernels per | Nitrogen in Kernels per | Nitrogen in Kernels per | Nitrogen in
plant. kernels. plant. kernels. plant. kernels.
Bl sg Boiog Bot g
gi cate 14
2 Ss = be -
36 | & é- |. 8 S A
= oO . ~_ iol oO — be fad) ~ me
Pee | Se Se | ee eel gt a | Be | Se eS
| fs A = = 5 = Eo =| 5 = oy
3 > 2 5 o = > o i <0) 5 5 i) 5 i)
va < = a = 4 <4 = ov = 4 < = ay =
Gms. Gms. Gms. Gms. Gms. Gms.
Cae et 555 | 10. 82 | 1.68 |0.1709 | 35....] 641 | 9.12 | 2.69 |0. 2491 | 112...] 1,052 | 17.71 | 2.68 | 0. 4606
655-77 654 | 14.66 | 2.20 | .3040 | 35....| 790 | 12.63 | 2.63 | .3262 | 99....] 1,040 | 19.13 | 2.84 | .5207
1Seeo-) GOL | 11. 26 | 2. 74‘) .3068 | 90....| 745 | 11.42 | 2.64) .2985 | 285...] 1,165 | 21.44 | 2.69 | .5716
1Otes=| 308 | 5.87) 3:27 | .1919 | 53....| 694 | 10.59.) 2.74.) .2947 | 180...) 1,057 | 19.49 | 2.82 | .5486
55ce 5. 306 | 6.44") 3.68 | .243] | 23....] 723 | 10.56 | 2.74 | .2875 | 51...-.] 1,093 | 23.16 | 2. 71 - 6154
Geese) e800 | 6:62 | 4.25 | -.2837 | 30.-..] 779 | 11.54 |-2.65 | .3137 | 98....] 1,245 | 22:02 | 2.80 | .6153
2.59 ) 2710 | 8ls..-| 1,183 | 20:12°| 2264 1 S25s7
Bee 274 | 5.68 |.5.13 | .2773 | 25....] 716 | 10.56
It is difficult to explain why such great variations exist when there
seems to be little or no tendency to transmit them. It seems ap-
parent that the variations must be due to differences in environment.
Since the ordinary factors of environment, as sunlight, warmth,
moisture, and apparent fertility of the soil, are constant for all
plants under our nursery conditions, we must conclude that there
are factors profoundly influencing the growth of plants beyond the
ordinary range of observation. :
Figure 2 shows the general appearance of the centgener nursery,
each centgener containing 100 plants 6 inches apart each way.
Figure 3 shows a single centgener just after growth has started in
the spring, about 40 per cent of the plants having winterkilled. The
great variation in the size of the remaining plants is probably due to
the effect of environment and is not hereditary. This environmental
variation is usually noted even in centgeners where most plants have
survived and is often interpreted as indicating real hereditary
differences.
A number of interesting problems are suggested. Why should one
plant, growing under practically the same environment as another,
collect from the soil two or three times as much nitrogen? Or why
should two plants yielding different quantities of grain collect the
same quantities of nitrogen? Table III illustrates these variations.
269
12 EXPERIMENTS IN WHEAT BREEDING.
TasLe III.—Data from six wheat plants, showing irregular variation in yield and in
nitrogen content of grain.
Kernels. Nitrogen in kernels.|| Kernels. | Nitrogen in kernels.
Plant = Plant
No = No. ae
= : a Total | 1 re otal
Y 117 oT ay Yay P > 19 > : ™
Number.| Weight. | Per cent. weight. | Number.| Weight. | Per cent. weight.
|
Grams. | Grams. Grams. Grams.
7A Le 1,058 22.879 2.45 0. 5605 || 2800522562 776 18. 507 3.57 0. 6607
ZITOS 3. = - 1,030 16. 679 2. 59 - 432451)" 23907. se 1, 167 23. 018 2. 86 | . 6583
i) Ona 927 | 16.026 1.74 | . 2789 || 24005... .- 1,495 | — 30.064 2.19 6584
The three plants, Nos. 21107 to 21109, are from the same mother
crowing in a single centgener, probably less than 2 feet apart, yet
the actual grams of nitrogen gathered differ more than 100 per
Fic. 2.—Centgener nursery, Nebraska Agricultural Experiment Station. Each centgener contains
100 plants.
cent. This difference is not inherited, as these plants rarely trans-
mit this quality. It therefore seems hard to explain on a difference
in the root development or in the functioning parts of the plant. As
plants growing only 6 inches apart commonly exhibit such differ-
ences, it can not be ascribed to a difference in soil solution. Differ-
ence in vigor of growth is not a satisfactory explanation, as plants
Nos. 23905, 23907, and 24005 illustrate. These three plants under
uniform conditions yielded different quantities of grain, yet the
heaviest yielder produced no more nitrogen than the lowest. Such
differences are not only common among plants from the same cent-
gener, but quite marked variations are also noted between cent-
geners from the same mother plant.
~69
>
i
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 13
Some of theresults obtained from thestudy of this problem are shown
in figures 4 and 5. Figure 4 is a plat of a single centgener (1907),
with the plants 6 inches apart each way, making the entire area 5
feet square. All plants in this centgener are from the same parent.
Each square represents a plant. Where no figures occur the plant
was missing. ‘The upper number shows the percentage of nitrogen,
the central number represents the kernels borne by the plant, and
the lower number the weight in grams of the good kernels. Each
plant was harvested separately, the kernels counted and weighed,
and the percentage of nitrogen determined. Two wave lines indicate
plants analyzing above 3 per cent nitrogen and one wave line those
Fic. 3.—Wheat centgener just after growth has started in spring, showing about 40 per cent of the
plants winterkilled. Note the great variation in size of the remaining plants.
analyzing between 2.8 and3 percent. A tendency to group is noted.
Those containing between 2.56 and 2.80 per cent are not marked.
One straight line indicates plants with between 2.55 and 2.40 per
cent of nitrogen; two straight lines, less than 2.40 per cent.
Figure 5 shows a section of the wheat nursery in 1907. The small
squares represent centgeners 5 feet square and the heavy lines out-
line family groups; that is, all the plants and centgeners within a
heavy line came from the same original plant. The percentage of
nitrogen was obtained by taking a composite sample from all plants
on the centgener. Variation is quite marked, although there is
some tendency for certain families to run high or low; as, for example,
family 339.
269
\
14 EXPERIMENTS IN WHEAT BREEDING.
An illustration of the irregularities in number and weight of ker-
nels, in percentage of nitrogen, and in total yield of nitrogen per
plant is afforded in Table IV, pedigree records of two families. The
wide variations were supposed at first to represent natural fluctua-
tions which would be in some degree transmitted, but the selection of
these high fluctuations has had no apparent effect in modifying the
mo
ony
ESS
1-0
| Ww
w Nv
od
N
N
oie
N
ity) -
0)
N
©
0)
2.48 ; 2.60
7/6 | $76 | 4/0 | $40
19.6 14.4 12.1 | 14.6 7.0
KV?
rapes
@ !
©
Fig. 4.—Wheat centgener of 100 plants, showing variations in yield of grain and of nitrogen in 1907. Upper
figures, percentage of nitrogen content; middle figures, number of kernels produced; lower figures, weight
of good kernels in grams. The various underscorings of the upper figures indicate five groups having
successively higher nitrogen content as follows: (1) Figures underscored with two straight lines lie between
2.15 and 2.40 per cent; (2) those underscored with one straight line lie between 2.41 and 2.55 per cent;
(3) those not underscored lie between 2.56 and 2.59 per cent; (4) those underscored with one wave line
lie between 2.80 and 3 per cent; (5) those underscored with two wave lines lie above 3 per cent.
nitrogen content of plants in a family, as there always seemed to be
a mean content for each family, to which the types returned,
As examples of variation, note that No. 35809 in family 42 has a
low yield of nitrogen, yet the yield of nitrogen found in its progeny
is practically equal to that of other members of the family. In
family 48 (Table IV) note that in 1903 the three plants selected
analyzed 3.82, 4.43, and 5.48 per cent of nitrogen, respectively,
269
:
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. ED
while the family as a whole contained 3.53 per cent. The progeny
of these plants returned to normal in percentage and total yield of
nitrogen, except No. 21909, in which the yield of grain was above
the average. Just why these wide fluctuations occur when every
precaution is taken to grow the plants under uniform conditions is
not very apparent.
In 1908 a more thorough investigation of this point was made.
Twenty-nine plants from the 1907 crop were all selected from a single
centgener, and therefore all came from a single plant in 1906. From
each of these 29 plants a centgener was planted, and also a row 14
feet long. The 29 centgeners were planted side by side, also the 29
rows. At harvest time all the plants in each of the 10 adjacent
2.6@ |2.65 3.041209 ( 2.7312.56 |2.59 1289) 2.77] 2.62) 2.55 42.77| 2.71) 2.55 | 2894248\2.77
fee sac lowrlan [set onebeee coer loin are adel eo
272 2.72|2.75 [2.71 |2.72 2.83} 2.63|2.58
«STE By aR i re Di ha Fae a
2.93 ]2.92|2.57 2.82 2.5313.00] 2.53|2.49 3.1) 2.68] 2.50|2.36
17 | as0|oa/fooe|ses|sec| se [xos|-r7 wr |seeleze|sse ese | = | 2
a
9
2.761249] 2.69] 2.59 | 2.63] 2.66 |2.95]255
Sa vende avs| 7 [ose ces coool
260] 316 [260] 3.02 |2.c6] 2.93] 3.06]2.67]275] 2.71 |2e9]202] 239 |263 Fe 6
: i ia
8713.17 |z.69|2.49|\2.6012.71 [2.48]2.60] 2 co|268]z.69|z2c8]292\269 2.68
385| 385]390] 39) | 407] 417| 417] s22}5ssIsse6
90 13.09 |2.79] 2.60 |2.89]2.15]277] 2.92] 3.00]2.77] 3.00]28312.83| 263 ex
0 | CaP i fed ora Fe
2.45] 2.63 |2.5¢ ]2.76 [2.ca] 2. 242.7) athe iee 2.79 (2. ae
fivelaae| aus [oaslass | casiaaalnt [ast nes wor bene
: 68] 200]2.68|2<9 |2 89] 2.66 |272 |3.02|2.73]2.76 |2eS] 2.68
2.63|2.4¢]2.682] 2.73 2.86 2.63)2.52|263]2.77| 2.73] 268|2.71 | 266
3
@
a3
o w
a
@®
eg oO
—- WwW
y »
on
® G
Nie Yio Yio YP
Ol allies Ole) OF
@w/~ WI of/% a
0
2.68
385
ft
9 5 408}425/4251555 |555 1 556|556] 3
i , : .19123442.66 |249] 265 | 268] 2.79|/287]2.90|2.62)3.16|284)272/1.76 | 268
314] 316 | 317 | 877 390 408) 408 556 |55逢] 3
2.7 3.20]2.73| 2.73] 2.72 |2.17]264/3-02] 2.66 |2.65]2.79|2.90]262/ 2.11 | 2.9
390/390] 408/408 Reali 556 |\556] 3
Fig. 5.—Diagram of a portion of the wheat nursery in 1907, showing variations in nitrogen content in cent-
geners and families. Each square represents a single centgener, and each area within heavy lines shows
the centgeners belonging to a single family. The upper figures represent the percentage of nitrogen;
the lower figures are the family numbers.
WwW N ® NIw Vin P/O Nig Vw ~v
CESUS SE aE Ge ak Pea
YO Vig NIB Vio nlw Mo DP
nv
0.)
oO
i‘)
10)
3°
nN
a)
Nn
Nig Mis » wo Vio nr
a %R o o gio 3
to
w N
&
a)
centgeners were harvested in order and analyzed. The 10 dupli-
cate rows were grown in a manner similar to field conditions; that is,
sown at the rate of 5 pecks to the acre and the rows 8 inches apart.
The plan was to see whether the same sort of variation would be
found among plants under field conditions. To secure a uniform
sample from the rows, 7 plants were harvested from each foot of
row, 98 plants being harvested and analyzed from each row. In the
centgeners the results were similar to those obtained in 1907. The
plants in the rows, being planted close together, yielded only about
one-tenth as much grain per plant, but the variation in yield and in
percentage of nitrogen per plant was even greater than in the
centgeners.
269
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17
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN.
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EXPERIMENTS IN WHEAT BREEDING.
18
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The 10
The rows
were more uniform in com-
position than the centgeners,
series.
but they show a variation in
duplicate rows are plotted in
end of the
as
imilar manner.
Also
there seems to be no con-
composition from 2.58 to 2.84
sistent relation between the
rows and the corresponding
:
jon
co)
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oa
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EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 19
plants. The variation in both centgeners and rows seems to be
due to local effects, and does not appear to be hereditary. We
may eliminate any possibility of hereditary effect by adding to-
gether the short rows in’ the centgeners which come end to end,
thus making 10 long rows with each centgener equally represented.
The results of such composite analyses are shown at the right of the
centgener plat, indicating a variation ranging from 2.40 to 2.81 in
percentage of nitrogen in the 10 rows. In the same way we may
divide the 10 original rows into blocks having each row equally
represented. Variation in the 10 blocks thus formed is shown, at
the bottom of figure 6, to be from 2.47 to 2.85 per cent of nitrogen.
RELATION OF YIELD OF GRAIN TO NITROGEN CONTENT.
Since some centgeners yield more grain than others, the 29 cent-
geners from the same parent plant, of which the 10 centgeners just
considered were a part, were arranged according to percentage of
nitrogen and summarized in groups of 5 centgeners. This summary
(Table V) shows some relation between yield of grain per centgener
and nitrogen content, the yield varying inversely with the nitrogen
content, but when the 29 corresponding rows were arranged in the
same way no such relation was shown.
TaBLE V.—Nitrogen content of 29 centgeners and corresponding rows from family No.
S351 in 1908.
? Arranged according to nitrogen in centgeners. Arranged according to nitrogen in rows.
_ — — —
& Z Centgener Corresponding a Row Corresponding | 8
S - row. a ; centgener. ' =
So a L | &
~_ FF = "
5 = gs ° is |e
: Do * — = ° : "> a =
S meres | be | : 2 ae on Re ete kek
= q 2 o 8 =| = ao hy = = I 2 i}|eS| ae
= oS See ee a 2 |8|>| 8] | fale
g 2 a es 2 a : = SN ie Goa eee ee = al
3 = =) > oe = ° = S = iS) = o | reals
, Z a = < 4 a 4 Z Z = Z eB |4° 124
: at oe =v
P.ct.| Grams.| Grams.| P.ct. | Grams.| Per cent. P.ct.|G@rms.| P. ct. | Grms.|Grms.| P. ct.
ae 2. 83 616 7.19 2. 67 194 eo ODsly On eee 2.78 | 189} 2.63 | 652] 7.52] 2.65
; oe ete 2. 63 614 (ell: 2. 66 200 2688 | bee cs 2.72 | 196 | 2.57 | 763] 9.03] 2.73
: Deteraisi< 2.56 851 9. 60 2. 65 173 BSOS | Oecens 2.65 | 194] 2.52 | 817] 9.14] 2.72
] Behan 2 2.52 767 9. 01 2. 57 179 La [as a ee 2.60} 195 | 2.57] 789 |11.13 | 2.62
: aes sc 2.44 799 9.01 2. 52 189 DHL ORS ee 2.55 | 188 | 2.55 | 777 | 8.72| 2.64
s ean | 2.34 916} 10.50! 2.64 209 2 687 | 4c 250°} 176 12550 |. 729 |8.50) | 268
‘ SSF
269
>
20 EXPERIMENTS IN WHEAT BREEDING.
TaBie VI.—Nitrogen content and yield of grain from 180 wheat plants, arranged in inverse
order of percentage of nitrogen, in groups of 10.
NINETY PLANTS FROM CENTGENER NO. 41801,
Plant No. cto Yield. Plant No. | she Yield. | Plant No. ng) Yield.
Per ct. | Grams. Per ct. | Grams. Per ct. | Grams.
A eae 3:44 1° A S5dGA eos: 8.62 | -UNS14 MGT. hoe ee Bee pe 7.49
7, Nei ca 32 OA BB ea ook 627] 46. S841 ap oS 2. 7a.
9 OES 3:21.) TRB Os. Sn wee ae 2 G2 tC AOLae Wl Bete cee a eee 2.43 6. 60
oe Cee Sa hat3.19-)) OA SGI) GaP aie. rene B.6l'-|, BSG Bas eee 2.43 7.39
er aes 3.19.4: : Gab SL, Go ee ies 2.61 1305-2627 ee 2.41 7.48
Ti gle ee i BPA ta Ui aie As ee 2.60 | '--7. 0041 Ca oxo. rlecoeee 2.41| 14.83
A” FE ore 8.131: © 4. R0GN Bee a oe ad 9. 60) Senta A ee 2.40 | 10.37
Tee a at 3.00 | 13-4 lao hee ee eee eee a eae ee 2 2.39 8:82
Bs fds $c at se 2.98 |- 19r6G Wel6. Loe Se tS BS )S a ah oe! ee Se 2.39 1.44
Bid: fie soot 2.98°| WebE a4. 8. 22) SSS Tae eee 2.39 8. 40
A verage..... 3.15 6. 52 Average. .... 2.60 8.87 Average... 2.41 7.99
lentes ewes 2.99.) = 2: A BB eas |. 2688 4° BOGE G8 ooo sare: oaeaee 2.37 8.00
I pe se cafe ok 999 130 88. BG es pe oe] 3.57 | +10. 5B BB cain tee 2.37 6.07 ’
eee dee 80) TORE se. as nee ete ae ee Se eee 2.36 8.30 $
aes ee 2.85 7.1 Teo ees oe 2.56 ee Sib, See eS © 2.34 11.07 4
y 7 RE a re ee 2.82 PFT, Teo aes eae, ae Ode |, oe Ne es ee 2.30} 9.59 ¥
Sane ae pee 2.82 ASI Oba s eee ae 2.51 |, G2. 26 Was woos 2.29} 13.16 ‘:
et See Se ae a te Ce a (\ 2250), Pr Bee. ol seh 2.29 8.19 .
ana erase 9.46) )\-) BaeMise heer ee eg [LMT Raat Set oe cere a 2.27 7.90 :
“(ite «Sains DAFT | TSE BOs Cte con 9:49) 6.16 1-565. Ceo eon 2.27 8.60 =
yee ee a os 3.974 ° 4 ARO). 2 ae S48.) 2.00" ll 44s) eee ee 2.27 6.85
Average... .. 2. 83 7.43 Average. 2.52 9.41 Average.....| 2.31 8.77
Sy BY A 2 710) PUSS, WO ie eae nee 2,48] 19:018496.-... Se eee 2.26 8. 66 =
APS Se cuit see 2.70 4+ 6; 8a Moa. ce tee 2.47 | 21.10 eee 2.25 9.85 1
axe emis “aie 2. 68-1 330, G0 Sa or ey 9:47 | 1218 1-30. eee 2.23 9.17
ih et Reo eae a 2-85. | ED Hoge a. eae ek 3 | — “S47 | 38°35: AB ce 2. 22 7.24
Woteria: Ne ceo he 9265 Bee Oe ole ne eel 2:48" 8d Gi s35 eo epee | 2.20 | 10.01
By haan ke tee 2.65} “A426: WAR, Fe So 7 cee es | "2346 | 19 O8 ADS 7iia. eee | 2.20| 14.78
ROSS BE 2.669): ABW ai eenes oe ee | “2246 }o SOT Bert: ea eee 2.20; 13.50
ER a 265 BOSe. WOOL E eeseek | 2G" 32 Ri TT. Aa eae ee 2.11} 10.31 .
Beha te bape © 65 |, 16-941). sce coe | 2.4k| B98 Bes... te eee 2.11 10.40
| ESD Ab 2 62.\9 Orit Ve Boia sabes p 249) (16, 20138." 3. =~ cee 2.09 6.90
Average..... 2. 66 9.47 Average..... | 2.46 ) 11.47 Average... .. 2.19 10. 08
NINETY PLANTS FROM Row No. 141801
Ps es Sea 8.31 a, ak: CAR ae} Te: | 2.77) 20.56 |
i oth sti Ls 3.29 EO eet, oe 2.76 | 1.69
Bho Asie 3.25 BO AB eR oie ee 2.76.) JA OB |
lp fo tek oe aa 3.20 {Sbupict os tou es 2.76 | 3.41
” ee ee 3.19 PiEOn ADar a eee ee 2.75 | 1.29 |
i, i gee ee oe 3.18 Ak WB ioe en 2.74) 2.51
1 SO ae Ree 3.18 Pay. eet pee 2.74 | 1.30 |
SRG Ne ae 3.17 4A ee tne 2.71 53
ee APES eee 3.10 co Re Pe a ee 2. 69 . 63 |
it Mp eoge 6 area 3.09 96 thad@. es. kee 2. 67 18 |
Average | 3.20 70 Average 2.73 1.32 |
3) Aa AS pie aye 8 3.09 30 NN BOLD osweace cotede 2. 67 65
1d RS oo en f 3.08 Ce. eee aA Se 2.66) 1.01
i etn SP nae ht Ag 3.07 ae RE 2.66; 1.21 |
eat ea ae 3.05 OB Tease ok etc s. ck. | 2.66] 1.29 |
yl ee ey 3.084." Sar ikGteuet cde. be 2.65] 7.76 ||
i ee ea Psa 3.00 721) BT ow ce xtienc ne | 2.64 .70 |
OIE tees ie Rak os 2 2.905). 2,27 WaBbas. acces Se 2. 64 45 ||
(Spe) shee RES Says 9.06) « 108 WER. ols : Tee .74 ||
oy Pak ah 2.90 198 A Os. ocece oka ok 2. 62 .57 ||
7) ea es SS Fa a 2.88") :3.02:1), BL goui sheen swans 2.61] 2.01 |
Average 3.00 1.27 Average..... 2. 64 1. 64
Ee ey ee 2.87 1 db A ic oehe ee 2.61 1.07
Se RE ee Seek a ee a 2.60} 1.18
MO: Situs oc aeeeu 2. 82 LOT BARE ko tare 2. 60 10 |
C2 Haris an oh 2.80 ORL > aaty necan ete 2. 60 94
EIA: seach Ss. 08k 2.80 7h Ow ee ee 2.58 99 |,
Te ee he 2.79 BS ih Bike, «tea seeks 2.58 .38
Fi gle 2 tee 2.90.1)" EE IP Ob, 5 satan tee 2.58 90
I Se See 2.77 BET Ss cn ceees 2.58| 2.50
ee 2.77 1 GET Boos stints ese 2.57| 2.16
SOU vo cwenec ait 2.77 .52 hee ica Sonate 2.56} 2.31
Average... .. 2.80 1,14 jj Average. . 2. 59 ) 1.25
269
3
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 31
Tasie VI.— Nitrogen content and yield of grain from 180 wheat plants, arranged in inverse
order of percentage of nitrogen, in groups of 10—Continued.
SUMMARY OF AVERAGES.
commonly varies as much as 500 per cent.
Groups from cent- Groups from row | Groups from cent- | re: roups from row
gener No. 41801. No. 141801. gener No. 41801. | No. 141801.
Group Group -
No. No.
. Grain per) x; Grain per . Grain per | »;; | Grain per
Nitrogen. plant. Nitrogen. plant, Nitrogen. plant. Nitrogen.) plant.
Per cent.| Grams. | Per cent.| Grams. Per cent.| Grams. | Per cent. | Grams.
ese. 2 3 3.15 6.52 +3. 20 OS RON uGiscte shied: 2 2.46 11.47 2.59 | 1.25
ETN cal 2 2. 83 7.43 3.00 17 4 eat ee eae 2.41 7.99 22D | 1.63
St oena: 2. 66 9. 47 2. 80 1d ee eae le 2.31 | 77 2.42 2.13
eect te 2.60 8. 87 2.73 13 71s | a ne 2.19 | 10. 08 2.00 | . 85
ae | 2.52 9.41 2.64 1. 64 | | /
The fluctuation in yield between plants within a centgener or row
To note whether this
wide variation bore any relation to nitrogen content, the results from
all the plants in centgener No. 41801 and from its corresponding row
were tabulated according to nitrogen content (Table VI). While
the summary shows a marked variation in nitrogen content, there is
no corresponding change in yield, although there is a slight irregular
tendency for yield to increase as nitrogen decreases.
While the foregoing data deal almost entirely with variations under
nursery conditions, it would be interesting to know whether such local
variations also occur under field conditions. That individual plants
vary in this way is without doubt true, as the records of the row plats
just cited show. To determine whether local variations were found
in field plats, two sets of data were secured in the following manner:
In one of the field plats a drill row 224 feet long was selected and
divided into 2-foot sections. The soil was of average fertility and
uniformity. The results are shown in Table VIII.
Also a plat of land 77 by 88 feet with a 5-foot margin outside of this
was sown to Turkey winter wheat, using a drill 5.5 feet wide. A very
uniform stand and quite uniform growth were secured. The plat
would have yielded about 30 bushels per acre. At harvest- time it
was divided into blocks 5.5 feet square, making 224 such blocks. A
composite sample was made from the harvest of each block and each
sample was analyzed for total nitrogen. Figure 7 is a diagram of this
plat, showing the yield of grain and percentage of nitrogen in each
block. The same variation is here found that has been noted hereto-
fore in the centgeners and nursery rows. For example, in the first
series of 16 blocks, Nos. 2 and 13 average 1.74 per cent of nitrogen,
while Nos. 3 and 8 average 2.07 per cent. This variation seems large
in view of the fact that each block has an average of 600 to 800 plants.
In this case the variation must be due to.some local soil condition.
When different numbers of blocks are grouped, the resulting areas are
of considerable size, as illustrated by figure 8. The plats shown at B
are 11 by 22 feet in area and each was sampled 8 times, yet they show
a variation in nitrogen content ranging from 1.81 to 1.97 per cent.
269
-
99, EXPERIMENTS IN WHEAT BREEDING.
REDUCING THE EXPERIMENTAL ERROR.
So great is the fluctuation in individual plants, due to variation in
environment, that there would seem to be no hope of improving the
Fablei ive ean
20 176| 160 12’ 12. 80
Bie aaa ara
223 191 12 ul 63 15
pa ae
222) 174, 142) 12 110)
189) 1.96 \492\/86 aos 486 \ 179 to oe
22) 205, 189] 173 'Al
00, 2.01 | .E9\ 477 (37 485 197 210)/ “
204; 188] 172 C)
- 8/3
x
XN
nt
BL 8h8
xO ly @ ly
NEN
@ lo 8 ln S fa
NEES S
ad a& o
S
N
x
-
BEEEEESS OCD S
2! 203 171 139 123 10 SI 7. 59 27 u
189| 2.// 199 187 186 |1-90| -90| .62| 18/|197|479|189
218} 202 170 “138 y 104 90 74 58) 42 26 10
217) “15a “137 12) 105 89 7. 57) 4! 2 3
pea age a
216 200 168 1 136 {20 10 88; 6 40 2: R
ieee agente
21s 199 i 151 1 19 10 8 7i 5 39 ws
190 194 194) 177, 189 186| 182] 187 | 180\/84\ /87\ 2Z04\/94\ 189
182) (66 150 i 118) 102) 86 0 4 8) 22 é
pag lem
213 197 18) 16S 149 133) 117 10! 85 69) 53 3
emgage
212 184 16 132 116| _100 52
PO reladg aes
2il 195 179 16 14 131 Hs
ra ee alee eee
210 178] 16 130) 50
438) 786 7 198) 189) 183|162|/96 199\199|206
0S i93 17 “161 a” ae 3 8!
Fic. 7.—Diagram of plat of Turkey wheat containing 224 blocks (each
5.5 feet square), showing the location of each block (lower figures) and
variations in the percentage of nitrogen in the grain (upper figures).
percentage of nitrogen
through the continu-
ous selection of high
fluctuates. Nocumu-
lative results can be
expected where the
error is larger than the
expected variation.
The problem of find-
ing a high-nitrogen
wheat seems to resolve
itself into the isolation
of pure strains and the
comparison of these for
nitrogen content. To
effect thisisolation and
comparison, a method
must be found that will
reduce error to the
minimum. Replica-
tion according to some
systematic method
seems the most prac-
tical way. A num-
ber of examples have been worked up from data at hand to illus-
trate the effect of replication in reducing error.
tN
%
FRCS
SSaNaS
ims
oe] eT ie
N
ST iat 8
Ss
IN]
§
Kd
i]
SeSms
i
NEN
S|
1aN
oT
x
.
N
xD
oy
492 £97 | 282 £86) 284) 493] 294)
SREHRERRGT AST Se
%
r
i
8
r
z
€
a
&
ed
re
A$
MRM: ee
Fig. 8.—Diagrams of plats of Turkey wheat, showing the arrangement of 224 blocks (each 5.5 feet square)
when combined in groups of adjacent blocks, with average nitrogen content for each group: A, Groups of
4; B, groups of 8; C, groups of 16.
REPLICATION OF SINGLE PLANTS.
In the first case the 840 centgener plants from the same parent,
heretofore referred to (fig. 6), were grouped in various ways. Starting
269
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 293
with the first plant and taking every forty-second plant thereafter
gave a composite group of 20 plants. Taking the second and every
forty-second thereafter in the same way gave a second group, and
so on in the same manner until 42 groups of 20 plants each had been
made. Ina similar manner 21 groups of 40 plants each were formed.
Also, each of the 10 centgeners, being 10 plants square, was formed
into 10 rows and the rows numbered 1 to 10. Each row would have
10 plants if the stand were perfect, but in this case it averaged only 8.4
plants per row. By combining the first rows, second rows, etc., in the
10 centgeners, 10 groups were made of 10 rows, or 84 plantseach. The
results of the above combinations are shown in Table VII. Where
20 plants, uniformly distributed, were combined, the variation in
nitrogen content was from 2.40 to 2.67 per cent. Where 40 plants
were combined, the variation ranged from 2.47 to 2.60 per cent, but
the 10 groups of 10 rows each varied only from 2.49 to 2.59 per cent.
Just what should constitute the limits of error in any case will depend
on the minimum limit of the variations which are to be detected.
In this case 0.1 per cent of nitrogen might be considered such a limit.
TaBLeE VII.—WNitrogen content of 90 plants of Turkey wheat from 1 centgener and of
840 plants variously combined into groups to show deviation from mean.
849 single plants variously combined into groups.
4
90 single plants in 1 centgener
(united when having same nitro-
Combination 1.—Four groups,
each composed of every 42d
C om bination
2.— Two
groups, each
composed of
Com bination
3.—A group
composed of
every 10th
row in 10
gen content). plant in 10 centgeners, or 20 every 21st Ph ees
plants. plant in 10 or 10 poo ane
centgeners, equaling a4
or 40 plants. plants.
ota ia aT ete . d ; g ¢ =
Sse} 8 her 8 = a a e S S =
e |3/ 8 > |8| 3 om 3 o 8 = 3 = 3
Reese Se Be) Bt Bee ee ee Boh ee ee
Zz |&| A Sethe 2 Oye A Z A Z A Zz A
Eick: Per ct. Per ch: Per Chi Per'ct: Per ct.
2.09) 1/—0. 482 2.58 | 4/+0.C008} 2.47 |—0.043 2.44 |—0.089 2.48 |—0. 044 2.49 |—0. 042
2.11] 2)— .462! 2.60 | 3)+ .028) 2.47-|— .043 2.51 |— .019 2.49 |— .034 2.51 |— .022
2.20} 3)/— .372 2.61 | 2+ .038) 2.48 |— .033 2.51 |— .019 2.50 |— .024 2.52 |— .012
2.22) 1j— .352 2.62 | 4+ .048) 2.50 |— .013 2.51 |— .019 2.52 |— .004 2.52 |— .012
2.23) 1)/— .342 2.65 | 5)+ .078} 2.51 |— .003 2.51 |— .019° 2.52 |— .004 2.53 |— .002
2.25} 1j— .322 2.68 | 2)+ .108) 2.51 |—..003 2.51 |— .019 2.53 |+ .006 2.53 |— .002
2.26} 1/— .312 2.70 | 1+ .128) 2.53 |+ .017 2.52 |+ .009 2.53 |+ .006 2.53 |— .002
2.27; 3)/— .302 2.71 1+ .138} 2.54 ;+ .027 2.59 |+ .061 2.53 j|+ .006 2.54 |+ .008
2.29} 2;)— .282 2.77 | 2)+ .198} 2.54 |+ .027 2.59 |+ .061 2.55 |+ .036 2.56 |+ .028
2.30) 1!— .272 2.78 | 1/+ .208} 2.58 |+ .067 2.60 |+ .071 2.59 |+ .066 2.59 |+ .058
2.34, 1!— .232 2.79 | 1/+ .218 ; ——_——
2.36] 1!— .212! 2.82! 2)+ .248} 2.513 0276 2. 529 0386} 2.524) .023 2.532 0188
2.37} 3|— .202/} 2.85 | 1/+ .278 <= = =
2.39) 3/— .182 2.89 | 1j+ .318} 2.46 |— .092 2.40 |— .128 2.47 |— .07
2.40) 1|— .172 2.92 | 2)+ .348} 2.51 |— .042 2.48 |— .048 2.48 |— .06
2.41) 2/— .162 2.98 | 2)/+ .408) 2.52 |— .032 2.50 |— .¢28 2.48 |— .06
2.43) 5|— .142 3.00 1j+ .428} 2.52 |— .032 2.52 |— 008 2.50 |— .04
2.44} 1/— .132)| 3.13 | 1]+ .558] 2.54 |/— .012 |} 2.53 |4 (o92|] 2.53 |— .01
2.46} 3|— .112)) 3.17] 1/+ .598} 2.54 |— .012 || 2.54/4 ‘979] 2.53 |— .01
2.47| 4/— .102|/| 3.19 | 2)+ .618} 2.54/— .012 |] 2.54/4 ‘o19| 2.54] .00
2.48) 2)/— .092 3.21 1}+ .638} 2.58 |+ .028 2.56 |4. 939 2.54 . 00
2.50} 1/— .072 3.22 | 1/+ .648| 2.64 |+ .088 2.56 | 4 "032 2.56 |+ .02
2.51] 1)— .062|| 3.44 | I)+ .868| 2.67 |+ .118 || 2.57 |4 ‘ogo | 2.60 |+ .06
2.53) 1)/— .042 -— —|| 2.57|4 ‘949 } 2.61 |+ .07
2.56} 1/— .012|) 2.572 214] 2.552| .0468/} 2.58 | 4 “gs5
2.57] 2/— .002 —| 2. 54 | 0364
2.528). 0365
bo
for)
©
94 EXPERIMENTS IN WHEAT BREEDING.
Where the plants were repeated 20 and 40 times the error was
almost what we might reasonably expect the actual variation in
pure strains to be, but where 10 centgener rows, or 84 plants, were
‘ combined the extreme error was within bounds. The data would
indicate that single plants would have to be replicated nearly 100
times to bring the variation within the limits of error.
REPLICATION OF 2-FOOT ROWS.
Table VIII Ulustrates the variation to be expected by replicating
2-foot rows. For these data a single 220-foot drill row in the general
wheat field was divided into 2-foot sections and a composite sample
made of each section. The sections were combinéd in two different
ways. The first combination was composed of every twenty-second
section, making 22 groups of 5 sections each, and the second com-
bination was composed of every eleventh section, making 11 groups
of 10 sections each.
Here, again, the extremes are rather wide, but if these are excluded
the results would be called satisfactory. If a comparison of pure
strains of wheat was being made under similar conditions, it would
be necessary to take for further trial the entire best half of the strains
tested in order to be within the limit of error. (See p. 30 and fig. 9.)
TaBLeE VIII.—Nitrogen content of 110 2-foot sections of drill row of Turkey wheat,
arranged in order of percentage of nitrogen and also in groups of 5 and 10, to show devia-
tion from mean.
Five rows in a | Ten rows inagroup
Single 2-foot rows (those with same nitrogen content united). group composed composed of every
of every 22drow.| llth row.
Nitro- Fre- Devia- Nitro- Fre- Devia- Nitro- Devia- Nitro- Devia-
gen. quency. tion. gen. quency. tion. gen. tion. | gen tion
Per cent. Per cent. Per cent. | Per cent.
1. 76 1} —0.271 2. 04 5 | +0.009 1.91 | —0.109 | 1. 96
1.79 1} — .241 2. 05 2/ + .019 1.96 | — .059 | 1.98
1, 81 1} — .221 2. 06 4| + .029 2.01 | — .009 ) 1.98
1.85 1; —.181 2.07 5] + .039 2.01 | — .009 2. 00
1. 86 2} —.171 2. 08 4] + .049 2.01 | — .009 2. 02
1. 87 1}; —.16l 2. 09 3 | + .059 2.02 | + .001 2. 03
1. 88 1; —.151 2. 10 6| + .0€9 2.02 | + .001 2. 04
1.89 1} —.141 2.11 1} + .079 2.03 |! + .O01] 2. 06
1. 90 Le) = 2481 2. 12 2] + .089 2.06 | + .041 2. 06
1.91 1} —.121 2. 13 2| + .099 2.09 | + .071 2. 06
1. 92 7); —.lll 2.14 3] +-. 109 2.09 | + .071 2. 13
1.93 5} —.101 2.15 1 | + .119
1.95 5 | — .081 2. 16 1} + .129 2.019 . 0355 2. 029
1.96 3| — .071 2.17 Ll + «138 =
1,97 2; — .061 2.19 2| + .159 1.96 | — .077
1.98 5} — .051 2. 20 3]; + .1€9 1.98 | — .057
1.99 2} — .041 2. 26 4| + .229 1.99 | — .047
2. 00 4); — .031 2. 28 ._ 1| + .249 1.99 | — .047
2. 01 7) — .021 2. 37 1 |}. + .339 2.03 |.— .007
2. 02 2/ — .011 |—-—— 2.03 | — .007
2. 05 6] — .001 22031 + .093 2.04 | + .003
2.05 | + .013
2.08 | + .043
2.09 | + .053
2.17 | + .133
2. 037 - 0443
weds ies
av aia.
ae
py o£
=
¢ f—
f
.
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 25
REPLICATION OF 16-FOOT ROWS.
Since 16-foot rows are frequently used as test plats, a determina-
tion was made of the variation in a series of these. One hundred
check plats from the 1909 field nursery were grouped by fives and
tens in the same manner as in the two cases just cited. The results
are shown in Table [X. The coefficient of variability and also the
extreme variation are less than in the cases just considered.
TaBLE I1X.—Nitrogen content of 100 16-foot rows of Turkey wheat, all from the same seed,
arranged singly in order of percentage of nitrogen and also in groups of 5 and 10, to
show deviation from mean.
Five rows in a |Tenrows ina group
Single 16-foot rows (those with same nitrogen content united): group composed composed of every
ofevery 20throw.| 10th row.
Nitro- Fre- Devia- Nitro- Fre- Devia- Nitro- Devia- Nitro- Devia-
gen. quency. tion. gen. quency. tion. gen. tion. gen. tion.
Per cent. Per cent. Per cent. Per cent.
1. 82 1 —0. 316 2.18 6 | +0. 044 2.09 —0. 056 2.10 —0. 037
1. 86 2 — .276 2. 20 1| + .064 7A | — .036 ya | — ,027
1.87 1 — .266 OLA 1|+ .074 2-13 — .016 AS — .017
1.89 1 — .246 Zee 4} + .084 2.13 — .016 OS — .007
1. 92 2 — .216 228 1} + .094 2h — .016 2.14 + .003
1, 93 2 — .206 Zao 2;}-+ .114 2.14 — .006 2.14 + .003
1.94 1 — .196 rH) 2) + .134 2.16 | + .014 2.15 | + .013
1.96 3 — .176 2. 28 2/}+ .144 2.17 | + .024 20.15 + .013
1.97 Z — .166 2. 29 3 | + .154 2.19 + .044 y Aa 5s + .013
1.99 1 — .146 Pew 3 | + .174 PAL + .064 2.18 + .043
2.00 3 — .136 2.34 2) + .204 j;—————_
2.01 1 — .126 2.35 1! + .214 2. 146 0292 251387 .0176
2. 03 5 — .106 2.36 1} + .224 = ee
2. 04 3 — .096 2. 38 1! + .244 2.07 — .058
2. 06 6 —. .076 2. 42 1 | + .284 2.08 | — .048
2. 07 Z — .066 2. 43 1} + .294 2EO9) | | —=s.038
2.08 6 — .056 2. 45 1| + .314 2.09 | — .038
2.10 3 — .036 2. 48 1| + .344 2.10 | — .028
2 AL 2 — .026 2.50 1 | + .364 2.13 | + .002
213 a — .006 PAs ve 1 | + .384 2.17 + .042
2. 14 2 + .004 ||————— ———_—_ 2.18 + .052
2ald 4 + .014 2. 136 . 1178 2.18 + .052
217 4 + .034 2.19 + .062
2. 128 . 0420
Where the 16-foot rows were repeated 10 times the extreme dif-
ference in nitrogen content was only 0.08 per cent. To further test
this question, 500 rows, each 16 feet in length, were planted in the
fall of 1909 under quite uniform conditions. These rows were har-
vested and combined in groups of 5, 10, 15, and 20, as in the previous
case. The results are summarized at the bottom of Table X. When
the rows are repeated only 5 times the error is too wide for satisfac-
tory results, but when repeated 10 times the error is small enough
for experimental purposes. Repeating 15 and 20 times gave only a
small further reduction in variation. |
A point of interest is the fact that in 1909, where rows were repeated
5 or 10 times, the experimental error was less than it was in 1910.
In fact, there is no way of establishing a set rule as to the number
of repetitions necessary, since the experimental error is influenced by
69826°—Bul, 269—13—4
26 EXPERIMENTS IN WHEAT BREEDING.
all the factors affecting the growth of plants, such as soil fertility,
climate, or insects. In order to know what this error is in a par-
ticular case it would be advisable to grow a sufficient number of
check plats in each system of plats to determine the error by actual
test.
Table X is a summary of results with the systematic repetition of
plants and rows. The column under “Coefficient ef variability”
shows that repeating 20 single plants in a systematic way has given
about as great accuracy in determining nitrogen content as 2-foot
rows repeated 10 times or 16-foot rows repeated 5 times. For deter-
mining comparative nitrogen content, repeating single plants 20 to
40 times in a systematic method seems to give quite satisfactory
results.
TABLE X.—Summary showing degree of error due to variation in environment, according
to several methods of comparison.
Number | Mean Average Standard | Coefli-
Classification. of nitrogen oe devia- | devia- ree os
groups. | content. d tion. tion. a
bility. =
Per cent. Per cent.
90 single plants (Table VII) .......-...-- 90 2.572 | 2.09-3. 44 10. 85
840 single plants (Table VII): ms a ial i ates ca
a 3 .47- t
Every 42d plant, 20 plants in a | b . 2 ee : ae = 2.32 ;
STOUD ES 2 we aeeas eee ee ees c . . 1.87 :
d 12 2.528 | 2. 40-2. 58 1.91
AVEIARC cae we eee ewe eee 10.5 2.531 | 2.442. 61 1.86
Every 2ist plant, 40 plants in a | { a 10 2.524 | 2. 48-2. 59 1.22
PYOUD! 3. oes cc-o Pee eee ee bill 2.540 | 2.47-2.61 137
Avverage:s.. a2 stese eet dae 10.5 | 2.532 | 2.47-2.60 1.50
As 10 centgeners, every 10th cent-
gener row, 10 rows in a group...... 10 2.532 | 2.49-2.59 1.03
110 2-foot rows, single rows (Table VIII). 110 2.031 | 1.76-2.37 5.35
Every 22d row, 5 rows ina group.... { 4 us - oo get. eo!
AVOISEOi: cco cteue os cn eee ll 2.028 | 1.94-2.13 2. 65
Every 1ith row, 10 rows in a group.. 11 2.029 | 1.96-2.13 2. 28
100 16-foot rows (Table IX) ....:........ 100 2.136 | 1.82-2.52 6.98
Every 20th row, 5 rows in a group... { b . . a: -- a. a iS
AVOTAQG. «iin dehnt es Paateees 10 2.137 | 2.08-2. 20 1.87 i.
Every 10th row, 10 rows in a group.. ae ES ase ~2.10-2.18. 10176 | -.0219 pea a
500 pa rows (in 1910): . 5a
ADUOWS Stace Sociale = We Gaels otoeen (oka ia eee 1.905 | 1.68-2.28 ;
a 25 1.904} 1.81-1.99 2. 46
Every 100th row, 5 rows in a group.. » = Dene . ae 9 3
d 25 1.8 1. 81-1. 98 2.42
A.VOTOBO. 00's ds eriswen tate led aR phe tik 1.905 | 1.82-2.00 2. 49
Every 50th row, 10 rows in a group.. { 4 e oe oe et
Every 33d row, 15 rows in a group a ry Seay es 1.905 | 1.86-1.96 1.45
Every 25th row, 20 rows in a group “s| pAtea'cin sek 1.905 | 1.85-1.96 1.27
269
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN. 27
THE SMALL-BLOCK TEST.
Figure 7 (p. 22) ulustrates the method of making the small-block
test and also shows the percentage of nitrogen in the grain from
each block. Table XI shows the result of repeating these blocks 4,
8, and 16 times in a systematic method, i. e., taking every fifty-sixth,
twenty-cighth, or fourteenth block (fig. 8). The experimental error
varied inversely with the number of repetitions, .but was only within
the limit of error when the repetition was 16 times.
TABLE XI.—Nitrogen content of Turkey wheat grown in 224 block plats (each 5.5 feet
square) in 1909 and 1910.
SYSTEMATICALLY REPEATED TO FORM GROUPS OF 4, 8, AND 16 BLOCKs.
Eight blocks in a group | Sixteen blocks in a group
Four blocks in a group composed of every 56th composed of every composed of every
block. 28th block. 14th block.
Nitrogen. Deviation. || Nitrogen. | Deviation. | Nitrogen. | Deviation. | Nitrogen. | Deviation.
Per cent. Per cent. Per cent. | Per cent.
1.78 —0. 128 1.81 —0.115 1.82 —0. 096 1.84 —0. 06
1.84 -— .068 1.85 — .075 1.87 — .046 1.86 — .04
. 1.85 — .058 1.87 — .055 1.88 — .036 1.87 — .03
1.88 — .028 1.87 — .055 1.90 — .016 1.87 — .03
1.89 — .018 1.90 — .025 1.90 — .016 1.89 — .01
: 1.90 — .008 1.93 + .005 1.90 — .016 1.90 + .00
| 1.91 + .002 1.93 + .005 1.92 + .004 1.90 | + .00
1.91 + .002 1.96 + .035 1.92 + .004 1.91 | + .01
1.93 + .022 1.96 + .035 1.94 + .024 1.9%) + .01
1.93 + .022 1. 96 + .035 1.94 + .024 1.92 + .02
1.94 + .032 1.96 + .035 1.94 + .024 1.92 + .02
1.95 + .042 1.97 + .045 1.95 + .034 1.93 + .03
1.98 + .072 1.97 + .045 1.95 + .034 1.94 + .04
2.02 + .112 2.01 +. 085 1.99 + .074 1.94 + .04
Av’ge..1.908 . 0439 1.925 0464 1.916 032 1.90 024
1.83 — .041 1.80 — .094 1.83 — 051
1.83 — .041 1.86 — .034 1.85 — .031
1.83 — .041 Les — .024 1.86 — .021
1.84 — .031 1.87 — .024 1.86 — .021
1.84 — .031 1.87 — .024 1.86 — .021
1.85 — .021 1.88 — .014 1.87 — .011
1.85 — .021 1.89 — .004 1.88 — .001
1.86 — .oll 1.90 + .006 1.88 — .001
1.87 — .001 1.90 + .006 1.88 — .001
1.87 — ..001 1.90 + .006 1.89 + .011
1.88 + .009 1.90 + .006 1.90 + .021
" 1.94 + .069 1.90 + .006 1.92 + .041
1.95 + .079 1.94 + .046 1.92 + .041
1.95 + .079 2.04 + .146 1.93 + .051
Av’ge. .1.871 . 034 1.894 . 0314 | 1.881 . 0231
28
EXPERIMENTS IN WHEAT BREEDING.
\.
» *
Taste XI.—Nitrogen content of Turkey wheat grown in 224 block plats (each 5.5 feet
square) in 1909 and 1910—Continued.
COMBINED IN GROUPS OF 4, 8, AND 16 meg BLocKs, TO SHOW THE EFFECT OF SIZE OF PLAT ON
ARIABILITY.
Four sets of 14 groups, with 4 adjacent blocks in each
group. blocks in each blocks in each
group. group.
Stand- | Stand- Stand- Stand-
iineon Devia-| ard || Nitro- | Devia-| ard | Nitro-| Devia-| -ard | Nitro-| Devia-| ard
wag tion. | devia- || gen. | tion. |devia-| gen. | tion. |devia-| gen. | tion. | devia-
tion. - tion. tion. ion.
Per cent. Per ct. Per ct Per ct.
179 (SO eee 1536: |=0-05, >|-2 - ee 0) 2. ST 0066 ee se 1.82 |—0.077 |.......
184: |= OB 7 Ae ae oe 1,587 \—=2 04. ee 1383 |=. (66 1s-2 455 1.85:1— 204874 -e
1. 86 -\—. 0487 |e ee LeSv i SOky eoeee es 1. Sh 26264522 ee: 1.85; |= 20487 42225 2
S84 — 027 ese ceSee 1; 88) 08 ele 15:88 31 —- S016 ees eee 1. 88 |. nOheids ee
191 5 008. |b S255 1.88 — 0S see es 1 88 OOF ae 1.89 |— .007 |......-
1 OV 322 Os: | es eee 1°89) =» 302 b... shes | BD: le DOS ieee =e 1.90 j}+ .008 j.......
108 Po ES le ‘cpg Ming hee Ss 9.90 1 2008 Js oS a! 1.90 |+ .008 [......-
1592 | OS ese ue 15901 — SOL, ie $926 4=' 02442-2520 1.90 |+ .003 j......-
108) | -eAO2S ee ae 1.91 Ox ( wile oe 1. SRA SO2A sc. 1.90 i= see
103" |e 2023 Ne stee s- 1.91 0. Mee aes 1592 t= s ORAL eee 1:91 J-- ..01S 1. Se 2
193 °\4=- F023 te aoe 1.95 |42°. 08 ie 1.92 |+ .024 |....... 1.93 |4-.. 083 [2
194 |---. 083 |. 2-228 195 202 ee 1. OSE 2 O34: =e 1.94 |+ .043 |,......
1.96: \4-. 05342. - £07 |= 06a) aoe 198 i4- 305¢.J5.. 1. 94: 1+ 30463 | .53-
1.98073). ten 198 ao Ove Ne See | 1.94 |+ .044 |....... 1.95 |-- . 068.12. 2--<-
Average. .1. 907 . 037 0. 048 1.91 . 0314] 0.0376) 1.896 0299) 0.0372) 1.807 0. 0363
182 |= 3072" sucess eiee eens ee 1:Sh. | Se. ce
18h)" 2 oo ee 181. |= 072: |b 1.8) 41—>. O81 S222
1.84 e052 Le eee L814 — 002 iE 1°37'|—=0. 021... See
1.85 es. 042 4 eee L8b 2 O72 he eee 187 |\— A021 Ik
185 |= 042. je. dete 1582) — S062: 52 sS5 1.88) |— S081 Io tcxces
1.88: SOU ee eee 16k |= VG ere 1-88: |—.. 01}: 22
RE Ee oe eS 1 of Al ee eh A apne, |S ee Sa a
LONE SOS. aoe. L871. .012 Nees 188 }|— 2OLT +]... 23255
eS) ei erg 0 be ia Pe LBS l= 1 O02 1s Scere 1: 86+] =. OLL 1... 2 oe
1 PL ec Sas | FS a a 1.89 }+ .008 |....... 1.90) | Ces ts Oe
LDA Ss ULS sleae eee 1: 809=— -O1S aS Saas. 1.90 |+ .009 |.......
WB PSA 2 Nee | Ls Oly |45.028 12 1.91 |+ .019 niger mag
1.94 |-+- . 048 |..222k.- 1:08 01- > O88 eee 1 PARIS roa 7-3 el (ee
1. 95-|4> 6058.2. eee 1.98 |+ .098 |....... 1.07 i> CRMs
1596" ==. 068) 1 ees 23038. 4-5 SI8- i. saees 200) 4-2. 108 toes
Average. .1. 892 . 0390 os 1. 882 . 0496} .0636) 1.891 . 0358} . 0492
269
.
Two sets of 14 groups,
with 8 adjacent
One set of 14 groups,
with 16 jacent
EXPERIMENTAL ERROR AND VARIATION IN NITROGEN.
29
TaBLE XI.—WNitrogen content of Turkey wheat grown in 224 block plats (each 5.5 feet
square) in 1909 and 1910—Continued.
SUMMARY SHOWING EXPERIMENTAL ERROR WHEN BLOCKS ARE ASSEMBLED IN VARIOUS WAYS..
Season of 1909.
Season of 1910.
a
=
S qd ' 1 ' _ [=| ' ' ' —
oh o {2 A= LS OF oO “es 4 = © a
Se Bt toe b| & E |S b
Classification. Ss | ee ee s j bse | o8 eo a dG jes
2a oq (A (eo | Bs] Ss og givgvo| 8s
he ao o Syd oO ao o eae os
v aS 9.9 wa | as | -a2 | ge £.2 wo | 8S lest
a g oo SsSicSia8 | ge ot seios | es
Assi 8° | 3°) er |) 881338) 2° | s*) er} Se
5 | @ K Bove tL Beh og i BS) oe
a |e SY 453 Z
PELLET CELLED Ce
Fic. 11.—Diagrams of plats of Turkey wheat, showing the arrangement of 224 blocks (each 5.5 feet square)
when combined in groups of adjacent blocks, with the average yield for each group: A, Groups of 4; B,
groups of 8; C, groups of 16.
when the blocks are repeated 4 times the range in yield varies from
an average of 595.2 grams per block to an average of 786.5 grams
per block, a difference of 32 per cent. If varieties were being tested
by the same system, this variation would be more than we might
expect in the yields of different kinds.
When the blocks are repeated 8 times the average variation ranges
from 627.5 to 717.8 grams, a difference of 90 grams, or 14 per cent.
By repeating 16 times the extreme variation is reduced to 47 grams,
or 7 per cent. However, with the exception of the extremes, the
variation here is small. The question now to consider is the mini-
mum number of blocks which will insure comparable results. If
comparable results are to be secured the first season, the blocks
should be repeated 15 to 20 times. If it were desirable to carry
the strains for a period of 3 years, repeating 8 to 10 times would
probably be sufficient, since this would give a total of 24 to 30 blocks
for the 3 years.
269
‘\
RELATION OF
EXPERIMENTAL ERROR AND VARIATION IN YIELD.
SIZE OF PLAT TO VARIATION.
39
It is very desirable in plant-breeding work to determine the
minimum size of plat that it is practicable to use, since with hundreds
of strains to try each year it would be impossible to handle them
in large plats.
Taking the above series of 224 small blocks, the
adjacent blocks could be combined to give a continuous series of
larger and larger blocks.
Figure 11 shows how these combinations
were made and Table XV gives statistical results, showing also in a
summary for two years the comparative effect of increasing the
size of the block and of repeating small blocks.
TaBLE XV.— Yield of Turkey wheat grown in 224 block plats (each 5.5 feet square) in
1909 and 1910.
SYSTEMATICALLY REPEATED TO FORM GROUPS OF 4, 8, AND 16 BLOCKs.
| Classification.
Single blocks
Every 56th block, 4
blocks in a group.
Average
q Every 28th block, 8
; blocksin a group. .
Average
Every 14th block, 16
blocksin a group. .
Four adjacent
blocks in a group.
Average
Eight adjacent
blocks in a group. .
Se oe
4
Average
Sixteen adjacent
blocksina group...
269
a
p=)
fo}
tH
oD co
S | Sa
E 2
POW tua:
Q Q
= aie
ss) oe,
7 Vo
2241680. 38
a #4/665. 82/613.
b 14/689. 96/645.
c 14/680. 95
d 14/683. 36
3. 21/459. 41/441.
2. 71/468. 01/433.
a 14/674. 20
b 14/686. 66
Season of 1909.
Extreme va-
riations.
373 -995
25-721. 25
75-786. 50
595. 25-728. 00
627 -744.75
Average de-
viation.
Standard de-
viation.
Coefficient of
variability.
eect:
81.98] 13
35. 79
yield
per block.
Mean
463.71
5. 35/468. 23/441.
5. 77|467. 09)412.
4. 42/450. 59)3¢7.
4. 69/468. 93)403.
Season of 1910.
Extreme va-
riations.
Average de-
viation.
Standard de-
viation.
300 -809
75-502. 7
25-504.
50-490.
75-581.
627. 50-704. 88
660. 38-717. 88
14|680. 43/653. 25-700. 38) 7. 24
former}
Voor}
ee
680
14/680
VARIABILITY.
529. 75-725. 25
617. 75-748. 25
624. 50-825
488. 25-801. 50
565. 06-775. 00
636. 13-829
586. 50-706. 75
611. 31-767. 87
603. 75-797. 06
37.36
30. 84
53. 14
70. 96
48.07
24. 57
55. 93
40. 25
35. 43
49.08
35. 75
61. 46
85. 80
58. 02
31.57
50.93} 7.42
48.87) 7.20
4.74,
70.30) 10.11)
1. 53/463. 721450. 86-490. 56
443.18
465. 30
504. 64
441.71
71/304. 31-538. 44] 32.65| 39.34
38-472. 50
38-515. 38
387. 25-497
389. 50-576 -
413. 75-587. 50} 31.84) 41.90
386. 75-493. 25| 26.37) 31.49
\398. 62-510. 25) 27.86) 37.97
894. 12-545 31.55} 40.01
‘396. 37-527. 62) 29.70) 38.99)
406. 69-509. 38) 21.67) 26.43,
Coefficient of
variability.
a ays %
62. 62} 13.50
17.88} 3.82
COMBINED IN GROUPS OF 4, 5, AND 16 ADJACENT BLOCKS, TO SHOW THE EFFECT OF SIZE OF PLAT ON
40 EXPERIMENTS IN WHEAT BREEDING.
TaBLe XV.—Yield of Turkey wheat grown in 224 block plats (each 5.5 feet square) in
1909 and 1910—Continued.
SUMMARY BASED ON THE AVERAGE OF BOTH SEASONS, SHOWING THE EFFECT ON COEFFICIENT OF VARI-
ABILITY OF INCREASING THE SIZE OF PLAT AS COMPARED WITH DISTRIBUTING THE SAME AREA BY A
SYSTEMATIC METHOD.
Coefficient of variability.
2 Four adja-
: Number of Plata cent blocks
Number of blocks combined in each group. groups increas Plats combined
averaged. ee distributed | and combi-
blocks systemati- | nation dis-
“as Zager times=16
blocks.
1 PRR tee eliotn an Pe ts ey tew a ote as EL ee 224 13. 25 13. 25
is hw oe eo Seek tes Set tt See Be eee eee ee 56 8. 48 5.44 3. 42
Beech Moe he hae Sack nee ce ae cia ee a 28 7.91 3.03
WO6dsc2 2s: = cto. 2 cow aie tect Reese oo eet ae eee 14 6. 45 1.80
SUMMARY BASED ON THE YIELDS OF 1909, ARRANGED TO SHOW THE RELATION BETWEEN SIZE OF PLAT AND
AVERAGE DEVIATION.
Baek i deviation _—
plats are made up by
Number} Total . i e
Shape of plat. of blocks | number | Average | Systematte method
in plat. | of plats. |2¢viation.
Every— Per cent.
Per cent.
Lio ou cerita Oa Soe oe een etal eo pee eee 1 9:76: |. 2 chmeues one ee
Uby 2.222 sees Bi taee 2 12 8.57 eee | ccs
1 Moy Cac: eeeow nn Uc aie ane ree ae 4 4
By Beat ce ee tae : 36 "3g |}56th block... . 5.05
2 by t eich Rae Ste ees a ee 8 . 6.05
V by GARE Se teat he. ae a, 8 28 6.08 |s25eh block... . . 2.96
Dhey Snad cede rapt aoe 5 ies ie Cee Aa het 16 14 5:30 | 14th block..... 1.53
By. BAe as pak oe a ee 28 8 5.37 |... > house ae
7by8 5.20. o's apn om arate ee ae
Starting with a coefficient of variability of 13.25 per cent, it is
decreased to 8.48 when the block is made 4 times as large, to 7.91 per
cent when 8 times as large, and to 6.45 per cent when increased 16
times in size. Table XV gives the result of repeating the same num-
ber of plats equal distances apart. Here we see that where the plats
are repeated 16 times the average variability for the two years rapidly
decreases to 1.8 per cent.
It might appear from a study of the first part of this table that if
the size of the plat were constantly increased the variability would be
constantly reduced. However, increasing the size of the plat beyond
a certain point does not continue to remove the cause of variability,
namely, variation in soil. The last part of the table, which contains
the data for 1909, is arranged to show the effect of increasing the size
of the plat. It indicates a rapid decrease in variability up to plats
16 blocks in size, but no decrease in the next two cases. While acre
plats are probably less variable than tenth-acre plats and tenth-acre
plats less variable than hundredth-acre plats, yet plats of this size
are too variable for direct comparison and they are much too large
269
9x
EXPERIMENTAL ERROR AND VARIATION IN YIELD. 4]
for practical plant-breeding work. On the other hand, repe eating
the plats in a systematic way constantly removes thé cause of varia-
tion as the number of repetitions increases. It then appears that the
most practical method of removing error is to repeat series of small
blocks a-rather large number of times.
CONSTANCY OF VARIATION ON THE SAME PLATS.
' Table XV gives statistical data for 1909 and 1910 on the same
blocks arranged in the same way both years. Figure 12 shows the
average yield of each section and the average percentage of nitrogen
in the two years for /909. 19/0.
sections a,6,c,andd. wezo W/TROGEN WELD NITROGEN.
The yield per block (SAMS oem 6a eee
varied about the same AF i's; BLK HAGE
SSRSRSMOSALe SS
for the two years, be-
ing highest in section c
in both seasons. The
variation in nitrogen
was not as regular,
section ¢ being highest
in nitrogen in 1909,
whilein1910therewas COLFFICIENT OF VARIABILITY.
mall but. redular COL eo. er eee
increase in nitrogen
mm--¢ to a. ~The
second part of figure 12
shows the coefficient
of variability in both
yield and percentage
of nitrogen when the
small blocks are com-
bined in sets Fig. 12.—Diagrams showing Turkey wheat grown in 224 blocks,
t of four combined in four groups (Table XV, a, b, c, d) of 56 adjacent
(Table XV). Section blocks to show variations in yield and nitrogen content in 1909
a was highest in varia- "71"
bility of both yield and nitrogen content in 1909 and low in 1910.
Section 6 was low in variation in yield in 1909 and high in 1910.
This would indicate that different seasons do not affect equally all
parts of the plat,’and illustrates the difficulty of ‘‘standardizing”’
plats by the system of sowing all plats to one crop for a season in
order to determine relative yield.
BAUCSRVRSFoALo
EF +-_ £ GF
£45 Za-za
6 2RBw
RIN
oe
gS
Zig
Gus
Gm
Ries
4
43r
Be
mil
Ol A
VARIATION IN YIELDS FROM CENTGENER PLATS.
The centgener method consists of planting 100 plants in a centgener,
6 inches apart each way, making blocks 5 feet square. In 1908, 178
centgener blocks were compared for variability with an equal number
- 269
49 EXPERIMENTS IN WHEAT BREEDING.
of duplicate row plats 16 feet in length. The variability was found
to be practically the same. Other data confirm this conclusion,
although under unfavorable ~conditions centgener plats are quite
variable, owing to the fact that the individual plants are so far apart
that the missing plants are not compensated for by the tillering of
neighbors, as is the case where the planting is at the normal rate.
ALTERNATING CHECK ROWS AS A MEANS OF OBTAINING COM-
PARATIVE YIELDS.
In order to test the value of the method of alternating check rows,
the 500 rows before referred to (pp. 36-37) were used as a basis for
data. It was assumed that every odd-numbered row would repre-
sent a check row, while every even-numbered row would represent a
strain being compared with a check row. Thus, row No. 2 would be
considered a strain to compare with rows 1 and 3 as checks. It is
apparent from data heretofore presented that the error would be too
great if only a single row were compared with its adjacent checks.
For example, there are numerous cases in the 500-row plats where the
even-numbered row would be 20 to 30 per cent higher or lower in
yield than the average of the adjacent odd-numbered row plats.
Table XVI shows the result of averaging five odd rows and the
adjacent five even rows, i. e., rows 1, 3, 5, 7, and 9 are averaged to
compare with rows 2,4,6,8,and10. Inthe first 10 rows, for example,
the five odd rows averaged 235.6 grams per row and the five even
rows 226.4 grams per row, or 9.2 grams less than their checks. In
the next block the even rows yielded 9 grams more than the checks.
Out of the 50 cases here cited the extremes vary from — 26.6 to
+ 32.8, with an average deviation of 10.14 grams or 4 per cent. In
selective work it is the unusually high variants that are sought after,
but with an experimental error greater than the expected variation
they would be difficult to locate. Table XVI also shows the result
of dividing the 500 rows into blocks of 20 and 40 rows and comparing
the yield of odd and even rows in each case. While in most cases the
average deviation is small, yet there are a number of quite wide
variations. For example, when 50 series of five odd rows are com- |
pared with five even rows, 17 series, or about one-third, show a
deviation greater than 5 per cent of the mean; when 25 series of 10
odd rows are compared with 10 even rows, 6 series, or about one-
fourth, show greater than 5 per cent average deviation; and when
12 series of 20 odd rows are compared with 20 even rows, none show
5 per cent deviation.
EFFECT OF INCREASING LENGTH OF ROW.
To increase the length of the row will decrease the error in about
the same way as to increase the size of the block. In the 500-row
plats just discussed most of the rows were in series and end to end,
269
Bul, 269, Bureau of Plant Industry, U. S. Dept. of Agriculture. ; PLATE I,
}
.
FiG. 1.—HEAD-TO-ROW NURSERY, IN WHICH 25 GRAINS FROM A SINGLE HEAD ARE
PLANTED IN A ROW 20 INCHES LONG.
The second year the seed from each 20-inch row is planted in a 16-foot row.
ie ert
aR eee ee ee ee yuo as i)
LUPE APOOREITTE Pa se CEP Ur,
PORMARRLRUSDLLL S mere yret ap ct. fe)
‘Erte ae leg ay Peeks
’
CBS 5
ripped y
Fic. 2.—ROW-PLAT NURSERY, IN WHICH THE ROWS ARE 16 FEET IN LENGTH WITH
A 4-FooT ALLEY ADJACENT, THUS MAKING THE BEDS 20 FEET IN WIDTH.
These beds are slightly rounded, to give perfect drainage.
Bul. 269, Bureau of Plant Industry, U. S. Dept. of Agriculture. PLATE II.
FiG. 1.—INCREASE PLATS OF ONE-THIRTIETH ACRE EACH.
Selected strains from the nursery are tested in these plats for 3 years.
Fic. 2.—INCREASE PLATS HARVESTED AND READY TO THRASH,
The plats in this field averaged 60 bushels per acre,
220. 6} 229. 6 9 | 225.6) 239.8) 14,2) 244.6) 230.4) —14,2) 222 226. 2 4.2| 242
EXPERIMENTAL ERROR AND VARIATION IN YIELD. 43
with only a narrow alley a few inches in width between the ends.
By adding together the rows end to end, longer rows could be made.
(Pl. 1.) A total of 84 rows 64 feet in length was made in this man-
ner, and the yields calculated. Table XVI gives the variability in
the original 500 rows, each 16 feet long, in comparison with the same
rows when combined into lengths of 64 feet. By increasing the
length four times the deviation and variability are reduced not
quite one-half. The longer rows are also less variable than blocks
of five adjacent 16-foot rows, but more variable than five rows dis-
tributed in a systematic way throughout the plat.
The best length of row to use must be determined by circum-
stances. If sufficient uniform land is available and it is more con-
venient to make long rows, to do so would lessen the number of
‘repetitions of plats necessary to reduce the error within proper
limits, but it would always take a larger area to secure the same
degree of accuracy with the long rows.
TaBLE XVI.— Yvreld, in grams, of Turkey wheat grown during the season of 1910 in
500 rows, each 16 feet in length.
ARRANGED BY ODD AND EVEN ROWS AND AVERAGED IN GROUPS OF TEN.1
{ |
Rows 1 to 100. Rows 101 to 200. Rows 201 to 300. | Rows 301 to 400. Rows 401 to 500.
|
Differ-| Odd. |Even.| Differ-
Differ- Differ- ‘Differ. |
Odd. | Even. Odd. | Even. Odd. | Even. od
214 | 231 FA be, 218 235 | 229 216 | 216 284 | 276
235 | 208 235 | 238 235 | 271 222 | 204 200 | 217
234 | 316 247 | 297 pe all 245 | 236 316 | 285
275 | 240 247 | 255 221 | 242 231 | 194 194 | 243
272 | 298 267 | 296 273 | 269 200 | ‘218 363 | 343
256 244.6) —11.4) 253.4] 245.6] — 7.8} 239.8] 244.6 4.8) 222.8] 213.6) — 9.2] 271.4) 272.8) 1.4
27 239 269 259 219 255 228 200 248 | 294
275 215 250 289 270 280 227 228 380 | 338
224 237 278 246 27. 191 218 208 326 | 287
220 240 222 256 282 249 242 257 265 | 315
182 263 308 276 268 237 221 230 314 290
234.6) 238.8 4,2) 265.4) 265.2) — .2| 262.8) 242.4) —20.4) 227.2) 224.6) — 2.6) 306.6) 304.8) -— 1.8
156 252 © 297 280 213 246 215 252 328 | 273
227 263 230 310 255 269 269 210 324 | 315
_ 254 224 323 263 235 242 171 228 300 | 279
~ 226 188 317 261 247 268 230 207 252 | 249
246 255 305 268 303 242 222 200 331 | 27%
221.8} 236.4) 14.6) 294.4) 276.4) —18 | 250.6) 253.4 2.8] 221.4) 219.4) — 2 |°307 | 277.8) —29.2
! Average difference in yield between odd and even rows, grams, 10.4; per cent, 4.
269 -
44 EXPERIMENTS IN WHEAT BREEDING. ee Sy es
TasLe XVI.—Yield, in grams, of Turkey wheat grown during the season of 1910 in”
500 rows, each 16 feet in length—Continued.
ARRANGED BY ODD AND EVEN ROWS AND AVERAGED IN GROUPS OF TEN—Continued.
Rows 1 to 100. | Rows 101 to 200. | Rows 201 to 300. | Rows 301 to 400. | Rows 401 to 500.
Differ- Differ- ‘Differ- ) Differ- iffer- 4
Odd. | Even. | Gnoe, | Odd. | Even. | ence. Odd. Even. | oxi Odd. | Even. “ence, Odd. |Even.” 08.
297 | 247 275 | 249 257 | 225 | 282 | 246 | 224 | 303
230 285 241 269 190 204 269 240 7 349 320
263 281 285 245 227 250 250 230 27 283
255 205 249 222 222 174 233 268 291 331
280 280 246 223 208 216 213 200 208 271
251 269. 6 18. 6} 259.2} 241.6] —17. 6] 220.8} 213.8) — 7 249.4] 236. 8 —12. 6} 268. 8} 301. 32.8
275 | 237 232 | 298 229 |. 187 200 | 207 269 | 203
264 P| 200 228 200 215 219 233 7! a
268 227 282 242 238 238 225 207 257 263
204 218 227 239 242 202 205 230 254 244
237 202 246 222 190 245 243 216 250 311
249.6} 232.2! —17. 4; 237.4| 231.8] — 5.6; 223.8] 217.4) — 6.4; 218.4] 218.6) Z| 264. 6] 287.6 23
—————d = “ | =—_—|-———
Pe el eet, 266 287 218 212 | 250 208 286 | 243
204 210 351 312 212 184 276 254 297 403
257 312 265 300 234 267 235 245 316 285
294. | 295 322 | 313 211 213 243 | 255 297 } 239 r
313 315 293 285 232 284 257 281 211 247 i
ashy aid Fae, * eS aI oh
260. 6} 272.2 11. 6] 299.4} 299.4 0 221.4) 232 10. 6} 252.2) 248.6) — 3.6] 281.4) 283.4 2
233 | 265 248 | 273 230 | 219 298 | 316 _ 208 | 219 e.
276 214 233 243 236 228 261 208 255 1 2565 ‘9
327 300 280 261 290 314 225 248 . 243 =| 289 $
BBL 316 259 263 208 195 233 226 247 275 3
370 290 248 Dis 225 219 205 214 249 266
303.6] 277 | —26.6] 253.6] 262.4 8.8] 237.8} 235 — 2.8] 244.4) 242.4) — 2 240. 4) 260. 20.4 *
271 236 241 250 232 211 “250 230 217 215
251 246 266 254 185 PSY 254 211 235 285 -
269 251 310 263 242 242 205 269 252 | 235 z
217 242 245 245 279 270 237 216 222 | 289 4
298 308 2f9 269 265 270 242 192 271 288 2 E FE |S >
ya fae ° O < ND me 4 Z
P.ct. |Grams.| Gms.| Grams. | P ct.|Days P. &.
TSG a He sank ome tite Eee 2. 66 13. 38 764 0. 02192 80 3 22 | 2.60
eT er eee EO re ne ey 2.52 12. 38 704 . 01864 81 35 23 | 2.49
a of tials gi yy in lee acpi or lanes eee . 2.68 11. 58 622 . 02105 63 35 23 | 2.48
BUG Mera. . fe CC comes see kek ae | 2.61 12. 27 646 »02344 68 36 31 | 2.55
WOO eas Mee. ses ae ee 2. 70 9. 83 612 01824 67 35 35 | 2.56
Total or average...... 2.63 | 11.89] 670 02058 | 72] 35 ) 134
ee RE om 2.72 | 12.36] 593 . 02267 55 36| 38
BR ce NS gta ne ala Ane wl teen eee 2. 53 11.12 623 . 01984 65 35 | 20
MUD Fs Oicnerd 6 Ja ha ven Rote ee eee 2. 63 11.14 605 . 02064 70 34/ 48
a ake oi ae ead Rao woraeca at ee ohne 2. 60 12. 26 664 . 02013 63 36 123
Be, Meee ts cade s aes he 2.70 11.01 620 . 02168 63 34 18
Total or average......... 2.64 | 11.58 | 621 . 02099 63 35 | 256 | 2.53
ee ed
1 Averages of check plats: Nitrogen content, 2.61 per cent; yield, 35.18 bushels.
269
EXPERIMENTAL ERROR AND VARIATION IN YIELD. 49
TABLE XX.—Relations of certain characters of 24 strains of Turkey wheat grown in
nursery and in field and tested during 4-year pertods—Continued.
RANKED IN GROUPS OF FIVE IN ORDER OF YIELD IN THE FIELD—Continued.
Data from field plats
(average for four
years, 1907-1910).
Data from centgener nursery (average for four years,
1906-1909).
a U n
Family No. 3 Yield per— cs : 5 “ 3
an = 3 |e0 oie
(The numbers in italic indicate = = ee ob 7 = 5 % € a
the five highest yielders.) 9 © of Ss S s gis - 5 r=) *
60 s 1 ao oO o— Ln 5D ° 2B
2 e = Bo 5 ae = A =
5 ee ee Pa de a ee
“4 a Ss) <* nD & | 4 z Ce 7,
P.ct. |Grams.|Gms.| Grams. | P. ct.\Days. | P.ct.| Bush.
SUle ob ooo eee 2. LO}, AOD93 10 SLO 02185 64 34 28 | 2.51 | 37.96 5
OPN 2665 CE ee 2.63 12.46 | 647 02295 64 34 59 | 2.59 | 36.57 12
CID. - no ee 2.50°| 11.95 4} 597 02118 68 36 114 | 2:53 | 36. 27 5
CNT oo CS 2. 81 11.05 | 563 02016 60 34 20 | 2.70} 36.13 5
OND. «cy jargon ee 2.59 | 11.08 | 558 01889 65 34 26 2.58 | 36.12 5
Total or average........- 2565) | 49e| eit 02111 64 34| 247| 2.58] 36.61 32
ZU 4 ieee ee eee 2. 68 9.70 | 560 02020 66 34 52 | 2.63. }) 35.22 8
WO oo ee 2.63 | 10. 70° | “512 02247 63 34 23 | 2.60 | 34.84 8
(Mh. 2 oe ee 2. 76 10.03 | 614 02029 66 34 18 | 2.73 | 34.20 5
>. toa Se 2.84] 10.79 | 620 02057 62 24 20, (12-40 eeanae 5
FINO ou Eo ee 2.02 || 12:00 | 594 02108 66 35 43 | 2.48 | 33.46 8
Total or average......... 2.69 | 10.67} 580 02089 65 34 | 156 | 2.63 | 34.26 34
AND on. ta S one eee ee 2.49 |) 11.38 | 570 . 01915 82 34 37 | 2.45] 32.93. 8
Bl nc Jo doe eS 2. 87 11.27 | 526 . 02270 54 34 48 | 2.75 | 32.58 5
2h Stee oe See ee eee 2.56 9.86 | 548 . 02180 58 36 33 | 2.55 | 31.88 8
BV Oso 3 See nee 2. 62 11. 89 508 . 02522 62 33 30 | 2.73 | 28.88 5
Total or average........- 2.63 | 11.10] 538 02228 64 34 | 148 | 2.62) 31.56 26
SUMMARY OF RESULTS, ARRANGED IN GROUPS OF FIVE STRAINS AND RANKED IN VARIOUS WAYS.
CENTGENER TESTS.
In order of nitrogen content:
Ghd! Ith 020; 000.02 ~~. - 2 2.80 | 11.10] 583] 0.02128 59 34] 144] 2.69] 35.07 25
425, 391, 3; 314, 42 Pe Pete ote 2.69 10. 61 585 . 02060 65 34 156 | 2.54] 38.15 49
48, 521, 168, 215, OSS Sassen 2. 63 TT e607 . 02224 68 34; 182] 2.61] 35.94 42
312, 47, 206, 25 295 psa San oe 2,58: | de 32} "608 . 02082 64 35 | 242] 2.55] 36.90 71
287 016, 313, 509 O08 Lee are 2.51 11.95 | 616 . 02001 74 35} 217 | 2.49 | 35.81 33
In order of ‘strength of straw:
209, 287, 48,215, 812.........- 2. 58 12.11 | 658 . 02076 76 35 161 | 2.53 | 38.42 46
313, 425, 314, 526, DING ok Sy. 2 2. 63 10.72 | 595 . 02020 67 35 262 | 2.59 | 35.67 34
200,225,091; 221 3.-.....-.- PAN os} 11.32 | 592 . 02104 64 34 160 | 2.55 | 37.58 55
ZANOS; 4% ,009,028----- 55. 23:67 11.44} 585 . 02189 63 34] 219] 2.61] 35.14 62
377,2, 556, OU es aaa eee 2:74}, TVA34 558 . 02183 57 35 139 | 2.63 | 34.86 23
In order of yield per plant:
48,221, 287 5000, O12; ..5----- 2.630) 27a Grek . 02192 i 35 Lis) 2.05) ,oor20 47
47, 216, 313, 328, 42 te Te 2.58 | 11.96} 597 . 02173 64 35 | 333 | 2.55 | 35:40 62
UO alsa, 229,200) 2.2. 005% 2. 62 11.20} 576 . 02024 67 34 188) 2:57 § 35. 8I 36
Si (i pSPOSE HOU) Gy! aa eee 2.76 10.76 | 585 . 02091 63 34 104 | 2.64] 36.07 43
168, 2 425, 314. 5h 5 2.64] 10.02} 558 . 02068 63 35 143 | 2.58 | 35.28 32
In order of yield per centgener:
Marea gAl, 221,012... ote 2.60 | 12.55.]° 685 . 02041 7 35. |. 258 | 2.55. | 39.12 81
995, 49'3, 379, 596.......-... 2.70 | 10.91] 620] .02068} 64] 34] 108] 2.60] 36.98 48
Yeo, 210,313, 216, 556...2..-.- 2.61 11.48 | 600 - 02076 65 35} 278 |. 2.53.) 37.28 34
209, 377, 314, 206, 76 SR ee 263 |e LOLOLsly 560 . 02040 66 34] 168 | 2.58 | 34.46 34
37, 168, 391, 398 ee cate © ates > 2.70; 11.20] 514 . 02306 61 34} 129] 2.65] 33.56 23
FIELD-PLAT TESTS.
In order of nitrogen content:
37,526, 328, 379,377..------- 2.78 | 11.01} 566 . 02179 61 34} 136 | 2.7 33. 07 25
; 14,78, 168,221,206. ........ 2.64] 11.46] 608] .02129| 68] 34] 182] 2.60] 36.70 42
425, 2, 225, 312, 556. Bee ee 2.62} 11.09] 604 - 02120 63 36 | 166} 2.55 | 37.58 40
215 3,313, 391, Al Sere 2.63 | 11.46 | 604 - 02120 66 35 | 331 | 2.52 |] 37.97 80
, 287; » 42, 216, 209 HS ene eereee 2.55 | 11.86] 622 - 01998 73 35 |} 126 | 2.47] 36.72 33
In order of yield per acre:
. YES 281 hoy Olay flO ne cn nese 2.63 | 11.89} 670 . 02058 72 35 | 184 | 2.54] 39.93 43
556, 225, 215, 47, eis sire, a cfs 2.64] 11.58} 621 . 02099 63 35 | 256 |} 2.53 | 38.65 85
3 391, 221; 313, 377, UO RS Cars = 2.65 | 11.49] 575 . 02111 64 34 | 247] 2.58] 36.61 32
314, 168, 526,379, 216........ 2.69| 10.67] 580] .02089| 65] 34] 156] 2.63] 34.26 34
, 2A SI) 20. ee ae 2.63 | 11.10} 538 - 02228 64 34 | 148 | 2.62] 31.56 26
}
269
50 EXPERIMENTS IN WHEAT BREEDING.
To sum up, the 24 pure strains have varied in the centgeners from
2.49 to 2.87 in per cent of nitrogen, from 54 to 82 per cent in strength
of straw, from 9.70 to 13.38 grams in yield per plant, and from 508 to
764 grams in yield per centgener. They also have shown a variation
Fig. 13.—Field plats of pure strains and check plats of original seed of Turkey wheat, 1910. The upper
numerals are family numbers; the lower, 4-year average yields. Two of the poorest yielders out of 26
strains came adjacent to two of the best. The difference in yield would not have been suspected from
the appearance of the plats.
Fia. 14.—Wheat nursery plats, showing variations in winterkilling. Pure strains were alternated with
the original Turkey wheat from which the strains were isolated. The original was mostly winterkilled
while many of the select strains withstood the winter well.
in average weight of kernel ranging from 0.01824 to 0.02522 gram. In
the field plats the percentage of nitrogen varied from 2.45 to 2.75 and
the yield per acre from 28.8 to 40.7 bushels—a difference of about 12
269
EXPERIMENTAL ERROR AND VARIATION IN YIELD. al
bushels. Some of these plats and the check plats noted below are
shown in figure 13. It is interesting to note that the check plats of
original unselected Turkey winter wheat averaged 35.18 bushels per
acre, or about halfway between the highest and lowest pure strains.
More strains surpass the check in yield than fall below it, but this is
probably because a large percentage of the poor-yielding strains were
discarded after the first field test in 1907. It appears that neither the
original selection of the 800 heads, nor the discarding of centgeners in
the nursery, nor the continuous selection of high-yielding plants within
the centgeners had any effect on eliminating the poor yielders. There
was a marked difference in the appearance of the pure strains, some
Fic. 15.—Field plats, showing variations in winterkilling between two pure strains of Turkey wheat.
Strain No. 377 isshown at the right and No. 102at the left; No. 377 withstood the winter almost perfectly.
having short grains and others long grains. They also varied in color,
lodging, and general appearance in the field, both in fall growth and
spring growth. Figures 14, 15, 16, and 17 illustrate these variations
better than they can be described.
Table XX also shows the data from Table XV grouped in series
of 5, and arranged in various ways to illustrate relationships. The
principal considerations in this work were the improvement of wheat
in nitrogen and yield. Records were kept of many characters of the
plant in the nursery, but evidence points to six that are of interest,
namely, (1) nitrogen content, (2) yield per plant, (3) yield per cent-
gener, (4) weight of kernel, (5) strength of straw, and (6) length of
fruiting period.
269
52 EXPERIMENTS IN WHEAT BREEDING.
PERCENTAGE OF NITROGEN.
The percentage of nitrogen is in inverse ratio to strength of straw
and length of fruiting period, but has no direct relation to other char-
acters. It is transmitted in the field plats as indicated in the summary
of Table XX. Astriking example of this conclusion is seen in a com-
parison of families Nos. 209and 37, Table XX. These familiesrepresent
the two extremes in percentage of nitrogen and strength of straw, with
an inverse relation, but are nearly the same in all other characters.
From 1903 to 1906, records were kept of individual plants selected
from the nursery. When these plants were classified according to
Fic. 16.—Increase rows of Turkey wheat, showing variations in the time of heading in different strains,
each from a single plant. Four rows of each strain are grown.
percentage of nitrogen or size of kernel, regardless of the family from
which they came, there was a marked inverse relation, the percentage
of nitrogen increasing as the size of kernel decreased. It seems prob-
able, however, that the individual plants having small kernels may
have suffered some degree of arrested development, since this relation
disappears when the pure strains are so classified. Percentage of
nitrogen and yield per acre in field plats vary inversely.
STRENGTH OF STRAW.
Strength of straw varies inversely with percentage of nitrogen and
directly with yield per acre and yield per centgener.
269
Bul. 269, Bureau of Plant Industry, U. S. Dept. of Agriculture. r*) PLATE III.
Fic. 1.—TYPE OF ROAD GRADER OR DRAG USED IN GRADING A NURSERY INTO BEDS
20 FEET WIDE TO AFFORD UNIFORM DRAINAGE.
Fic. 2.—GRAINS OF TURKEY WHEAT, SHOWING VARIATION IN APPEARANCE.
Nos. 51 and 60 are typical kernels from two pure strains and represent the shortest and longest
kerneled types out of 80 strains. No. 76 isa hard, vitreous kernel, somewhat approaching
the durum wheat in type. No.75isa soft wheat. The plants of thisstrain are typical Turkey
in appearance but the grain is larger and almost white. Notwithstanding the white color,
this strain was the highest in nitrogen content of 80 strains in 1910.
Bul. 269, PLATE IV.
Fic. 1.—REPRESENTATIVE KERNELS FROM 4 STRAINS. OF TURKEY WHEAT, SELECTED
TO SHOW VARIATION IN APPEARANCE.
No. 48 is a large plump-kerneled strain, while No. 287 has a rather small kernel. No. 328 has
a large dark-colored kernel, while No. 3138 hasa decided yellow colorand is long and pointed.
Fic. 2.—REPRESENTATIVE KERNELS FROM 4 STRAINS OF TURKEY WHEAT, SELECTED
FROM A SERIES OF 80 STRAINS TO SHOW VARIATION IN QUALITY.
On the basis of a perfect wheat, grading 100, these strains grade as follows: No. 77 grades 50,
No. 51 grades 70, No. 27 grades 80, and No, 42 grades 95.
EXPERIMENTAL ERROR AND VARIATION IN YIELD. 53
YIELD PER PLANT.
The yield per plant shows some correlation to yield per acre and
yield per centgener, but this correlation is not high, as it is only in the
first class that the correlation is marked. One of the best-yielding
strains (No. 425) had a small plant yield in the nursery.
YIELD PER CENTGENER.
The yield per centgener shows a high correlation with yield per acre
and strength of straw, but not a close relation to other characters.
SIZE OF KERNEL.
The size of kernel (PI. ITI, fig. 2) appears to have no fixed relation-
ships; as a character of a pure strain it seems to be independent of
other characters. An exampleof this is shown in Table XX. Families
Fig. 17.—Field plats of Turkey wheat, showing variations in stiffness of straw in two strains. Each strain
originated from a single plant.
Nos. 287 (Pl. IV, fig. 1) and 425 have small kernels, but they are
among the best in yield, while No. 328 (Pl. IV, fig. 1) is poorest in
yield, but has the largest kernel. Nos. 48 and 287 are the best
yielders out of the 26 strains (Table XX), averaging 40.7 and 40.6
bushels per acre, respectively, in a four-year test. No. 48 has a large,
plump kernel, while No. 287 has a rather small kernel. No. 328 has
averaged 28.9 bushels under the same conditions, yet this strain has
a large, dark-colored kernel. Our records do not seem to show a rela-
tion between the appearance of the berry and the yield. No. 313 has
averaged 36.3 bushels per acre, but the kernel has a decided yellow
color, and is long and pointed in shape, approaching a rye grain in type.
QUALITY OF KERNEL.
As already noted, there does not seem to be a definite relation
between the appearance of the berry and the yield of the strains.
269
54 EXPERIMENTS IN. WHEAT BREEDING.
Plate IV, figure 2, illustrates four strains of Turkey wheat selected
from a series of eight strains to show variation in quality. On the
basis of a perfect wheat grading 100 these strains grade as follows:
No. 77 grades 50, No. 51 grades 70, No. 27 grades 80, and No. 42
grades 95. ‘These grades indicate the variation in quality found in
pure strains and show the great possibility of improving quality
(fig. 18).
To sum up, high yield in the field is associated with high yield per
centgener and strong straw, has a slight relation to size of plant, no
relation to size of berry, and varies inversely with percentage of
FIG, 18.—Cereal laboratory, showing the method of taking notes on quality. Comparisons of 80 strains
of Turkey wheat are being made. There were 10 plats of each strain, making 800 in all, but the 10 samples
of each strain are arranged together. Notes are taken on each sample separately, then an average is made
of the results. To facilitate note taking, a set of ‘‘standard samples’’ representing different qualities is
kept in jong, 2-ounce vials. A set of these vials is plunged into the sample, and by comparison very
accurate data are obtained.
nitrogen. A high or alow nitrogen content as indicated in the nursery
gives correlated results in the field. High nitrogen content is antag-
onistic to high yield: However, an occasional strain, such as No. 48,
combines a fair percentage of nitrogen with high yield.
SUPERIORITY OF STRAIN.
From the point of view of yield per acre there would seem to be
little choice among the five best strains (Table XX), but in some
ways No. 48 is outstanding in desirable qualities. With a high yield
269
EXPERIMENTAL ERROR AND VARIATION IN YIELD. 5d
per acre, it still is above the average in nitrogen content, has a strong
straw and a large berry of good appearance. It is also very winter
resistant, as was shown in the severe winter of 1909-10, when it
came through with much less winterkilling than standard varieties,
such as Big Frame and ordinary Turkey wheat.
COMPARISON OF ROWS, CENTGENERS, BLOCKS, AND FIELD PLATS.
In 1909-10 the 26 strains of Turkey wheat were sown in field plats
and duplicated in rows, centgeners, and blocks, but this portion of
the nursery was winterkilled. However, very good results were
obtained with 11 varieties of oats sown in the spring of 1910 in all
four ways. The field plats were one-fifteenth of an acre in size and
Fic. 19.—Block nursery, showing blocks 4.2 by 16 feet in size. The beds are slightly elevated, as in the row
nursery.
were repeated three times. Each variety was repeated 10 times in
centgeners, rows, and blocks. The centgeners were each 5 feet square
and contained 100 plants 6 inches apart each way. The rows were
12 feet in length and the grain sown in them at the rate of 10 pecks
per acre, the usual rate of seeding in this region. The blocks were
each 4.2 by 16 feet, or 5 drill rows wide (fig. 19), and sown at the
usual rate of seeding. The results summarized in Table XXI show a
high degree of correlation between the yield of the field plats and the
rows and blocks, but practically no correlation in the case of cent-
geners. Except for the Lincoln oat, which yielded exceptionally
high in the nursery, the correlation in the rows and blocks would be
very high.
269
56 EXPERIMENTS IN WHEAT BREEDING.
Taste XXI.— Yields of grain from 11 varieties of oats grown in field plats, centgeners,
rows, and blocks.
[The field plats were repeated 3 times, the others 10 times.]
a ————————————
Average yield of thrashed grain.
Variety. Field
plats, cane Rows. Blocks.
per acre. 8 ¢
Bushels. Grams. Grams. Grams.
60. 7 729. 2 Pe
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82 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
A number of other facts were observed which have not been
incorporated in the table. Thus it was found that in the cultures
on Raulin’s medium the tartaric acid soon disappeared. At least
none could be isolated as the acid potassium salt. Indeed, the
tartaric acid seems to be attacked at the beginning more rapidly
than the sugar. Pfeffer’ has reported analogous observations upon
Aspergillus. Reichel? found very recently that Penicillium when
grown in the presence of acetic acid begins to destroy the acid until
the unfavorable acidity is reduced. A concentration of acetic acid
equivalent to the tartaric acid of Raulin’s medium was found to
inhibit the development of Penicillium puberulum. That is why
in one of the experiments of Table IV sodium acetate was used.
There seems to be a general tendency for molds to reduce the acidity
of a very acid medium. This may be done by destroying the acid, or,
if the acid can not be attacked, by neutralizmg it with ammonia
when this can be formed by deamidization.* At any rate in the
present instance the sugar disappeared more slowly than. the tartaric
acid. By the end of the fourth week, however, the medium was no
longer optically active‘ or fermentable. Because penicillic acid ren-
ders the solution antiseptic the fermentation test is not in this instance
reliable. Nevertheless the medium reduces Fehling’s solution, 25
cubic centimeters yielding 78.1 milligrams of cuprous oxid.? The
reduction is caused by penicillic acid.
Alcohol was determined by taking the specific gravity of the recti-
fied distillate. The cultures contained 0.1 per cent as early as the
end of the first week. The low concentration of alcohol found
might not be the result of a scanty alcohol formation, but of the
further oxidation of the alcohol formed, since alcohol is a good food
for Penicillium.’ Indeed, it has been said that “P. glaucum” does
not produce alcohol at all.7_ Dox states that alcohol of a concentra-
tion not over 0.1 per cent is produced only when the air supply is
insufficient. All cultures of Penicillium puberulum tested con-
tained alcohol, even those grown in the flat bottles above described,
in which the aeration was certainly good.
1 Pfeffer, W. Ueber die Election organischer Niihrstoffe. Jahrbiicher fiir Wissenschaftliche Botanik,
Bd. 28, p. 205-268, 1895.
2 Reichel, J. Ueber das Verhalten von Penicillium gegentiber der Essigsiiure und ihren Salzen. Bio-
chemische Zeitschrift, Bd. 30, p. 152-159, 1911.
8’ Butkewitsch, Wl. Umwandlung der Eiweissstoffe durch die niederen Pilze im Zusammenhange mit
einigen Bedingungen ihrer Entwickelung. Jahrbiicher fiir Wissenschaftliche Botanik, Bd. 38, p. 198, 1902.
4 The determinations were very kindly made by Dr. C. 8. Hudson, of the Bureau of Chemistry.
5 This determination was very kindly made by Dr. H. Hasselbring, of the Bureau of Plant Industry.
6 Hasselbring, Heinrich. The carbon assimilation of Penicillium. Botanical Gazette, v. 45, p. 170-193,
1908.
7 Brefeld, Oscar. Ueber Gihrung. III. Vorkommen und Verbreitung der .\lkoholg#hrung im Pflanzen-
reiche. Landwirthschaftliche Jahrbiicher, Bd. 5, p. 315, 1876.
§ Dox, A.W. The intracellular enzyms of Penicillium and Aspergillus, with special reference to those of
Penicillium camemberti. U. 8. Department of Agriculture, Bureau of Animal Industry, Bulletin “
p. 33. 1910, ’
270 ,
PENICILLIUM PUBERULUM. 33
No glycerin could be detected in the culture fluid when 200 cubic
centimeters, rendered weakly alkaline with sodium carbonate, were
concentrated to a sirup, the sirup extracted with alcohol, and the
alcoholic extract after evaporation tested with the borax bead.
Though the Raulin medium cultures remained distinctly acid to
litmus, no nonvolatile acid other than penicillic acid could be ex-
tracted with sulphuric ether, petroleum ether, or acetic ether, even
after acidifying with phosphoric acid. No insoluble lead, copper,
calcium, barium, or zinc salt could be obtained. Great care was
taken to detect oxalic acid, but in young cultures none could be
found. Even in old cultures none could be detected by the ordinary
method of extraction with ether. A small amount was isolated in
the following manner: Seven hundred cubic centimeters of culture
medium about two months old were concentrated to a sirup, acidified
with phosphoric acid, and mixed with clean sand and plaster of Paris.
When the plaster had set, the mass was ground and extracted with
ether in a Soxhlet extractor. Oxalic acid, identified by its melting
point, crystallized from the extract, which also contained other
material, as shown by the evolution of gas and the odor of nitrous
oxid. Apparently some nitric acid passed from the medium into
the extract and there caused decomposition. The extract contained
a substance soluble in chloroform and giving a bright-green color
with ferric chlorid. An alkaline solution of penicillic acid when con-
centrated to a sirup does not yield oxalic acid, but it does give a
green color with ferric chlorid. Fumaric acid was absent. As
already indicated, the culture medium contained unidentified sub-
stances. This was further shown by the fact that by the method of
Griess, using sodium nitrite, sulphanilic acid, and acetic acid, an
azo dye of a beautiful carmine color was produced. This reaction
was obtained by Raciborski! with a number of fungi.
Only very small quantities of volatile material other than alcohol
were detected in the culture medium. For the purposes of this
examination a culture medium from which the penicillic acid had
been removed as thoroughly as possible with chloroform to avoid
obtaining its decomposition products, was used. The medium was
then distilled and the distillate extracted, first with chloroform and
then with ether. The residue from the chloroform consisted of a
few small white crystals with a melting point of 112°C. The residue
from the ether consisted of a few fine hairlike crystals. None of the
erystals gave the ferric-chlorid reaction. Distillation of this liquid
with mineral acid yielded no other products. Distillation with
1 Raciborski,M. Uber die Assimilation der Stickstoffverbindungen durch Pilze. Bulletin International
de l’Académie des Sciences de Cracovie. Classe des Sciences Mathématiques et Naturelles, ann. 1906. pb.
733-770. 1907.
270
84 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
alkali yielded a few white crystals obtained by extracting the dis-
tillate with ether.
All these experiments refer to the unmodified Raulin’s medium.
The mycelium was also studied. From mycelium of varying age
grown under different conditions no toxic material could be ex-
tracted with boiling alcohol. An oily, waxy residue remained after
the alcohol was removed on the steam bath. When this residue was
extracted with water and the extract injected subcutaneously into
mice no serious toxic effects were observed.
Mannitol has long been known as a constituent of Penicillium.
Trehalose and trehalum, or substances resembling them, have also
been described. It is possible that in fungi some investigators may
have mistaken trehalum for starch or glycogen.? Cramer *® found
that by treating the spores of Penicillium with boiling water no
carbohydrate material precipitable by alcohol passed into the extract.
When, however, the spores were exhausted with ether before the
extraction, a carbohydrate was obtained giving a deep blue color
with iodin. This carbohydrate, Cramer thought, resembled hemi-
cellulose, but it may have been similar to trehalum. Moreover, a
direct relationship between mannitol, trehalose, and trehalum has
been demonstrated, trehalose being formed only at certain stages
of growth, while later only mannitol occurs.’
Mannitol was readily detected in the mycelium of Penicillium
puberulum by extracting the mycelium dried in air with boiling
alcohol. On cooling, sweet, fine, white, silky needles separated, sol-
uble in water and alcohol, insoluble in chloroform, and with a melting
point of 162° to 163° C., uncorrected. Cholesterol reactions were
negative. They did not reduce Fehling’s solution, though they did
so after oxidation with nitric acid. They rotated polarized light
slightly to the left.
In the mycelium dried in air neither trehalose nor trehalum could
be detected. When fresh mycelium was immersed in boiling alco-
_ hol, as soon as removed from the culture flask and the boiling extract
filtered, no trehalose separated on cooling. This extracted myce-
lium, boiled with water and filtered hot, gave, on cooling, a small
quantity of gummy material which iodin colored intensely violet
and which was not easily inverted by hot dilute hydrochloric acid.
This substance is plainly trehalum, in no way mistakable for glyco-
gen or the more readily soluble and easily inverted starch.
1 Zopf, Wilhelm. Die Pilze, Breslau, 1890, p. 125.
2 Lippmann, E. O. von. Die Chemie der Zuckerarten, Aufl. 3, Halbbd. 2, Braunschweig, 1904, p.
eon! E. Die Zusammensetzung der Sporen von Penicillium glaucum und ihre Beziehung zu der
Widerstandsfiihigkeit derselben gegen iiussere Einfliisse. Archiv fiir Hygiene, Bd. 20, p. 197-210, 1894.
‘Lippmann, E.O. von. Op, cit., p. 1427.
270
PENICILLIUM PUBERULUM. 85
Both the culture fluid and the mycelium were examined for oxi-
dizing enzyms. The former contains an abundance of catalase,
though no oxidase detectable by guajac, aloin, or benzidin. A very
faint peroxidase reaction was found, due perhaps to the presence of
chlorids.: The statement of Loew ? that filtered Penicillium glaucum
cultures contain only catalase is therefore amply confirmed. Fresh
and air-dry mycelia were ground in a mortar with distilled water and
allowed to digest at room temperature for several hours. The
extract, filtered through paper, contained far more catalase than
the culture medium, but neither oxidase nor peroxidase could be
detected by the color tests. In performing these tests great care
was taken to vary the reaction, for this has been shown to influence
these tests greatly.
The mycelium was then tested for oxidizing power by the method
of oxygen absorption as developed in this laboratory by Dr. H. H.
Bunzel.* The air-dry mycelium was ground and the dry powder
obtained used directly in the oxidase apparatus in the presence of
pyrogallol and of tyrosin. No oxygen absorption was observed. To
make certain that neither the drying nor the acid of the medium was
accountable for the negative results, the organism was grown on a
medium containing monosodium phosphate and disodium phosphate,
which, as shown by Henderson and Webster,® remains neutral. This
medium was Raulin’s solution, to which was added 5 per cent of a
mixture of two parts Na,HPO, and one part NaH,PO, On this
medium few spores developed in 10 days. The mycelium remained
colorless. The medium contained alcohol and only traces of peni-
cillic acid. Ten grams of perfectly fresh mycelium of 12 days’ growth
were ground with clean sand and then transferred to the absorption
flask, together with 4 cubic centimeters of 10 per cent pyrogallol.
This concentration of pyrogallol was selected because of its antiseptic
action. Under these conditions no oxygen absorption was observed
for more than an hour. Absorption then gradually began. This
phenomenon is still under investigation.
Some of the results obtained with the variously modified Raulin’s
medium require further comment.
In the distillate from the succinic-acid culture aldehyde was
detected by the power to reduce ammoniacal silver, and acetic acid
by the formation of the ethyl ester with its characteristic odor.
1 Alsberg, C. L. Beitriige zur Kenntnis der Guajak-Reaktion. Archiv fiir Experimentelle Pathologie
and Pharmakologie, Festschrift Schmiedeberg, Supplementband, p. 39, 1908.
2 Loew, Oscar. Catalase, a new enzym of general occurrence, with special reference to the tobacco plant.
U.S. Department of Agriculture, Report 68, 1901.
3 Alsberg,C. L. Op. cit.
4Bunzel, H. H. The measurement of the oxidase content of plant juices. U.S. Department of Agri-
culture, Bureau of Plant Industry, Bulletin 238, 1912.
6 Henderson, L. J., and Webster, H. B. The preservation of neutrality in culture media w ith the aid of
phosphates. Ji ournal of Medical Research, v. 16, p. 1-5, 1907.
270
86 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
The observation made on the 1.5 per cent ethyl-aleohol culture’
that mycelium failed to develop from spores, whereas inoculation
with pieces of mycelium resulted in growth, is in harmony with the
statement of Duclaux ! that while alcohol restrains or arrests the ger-
mination of mold spores it is utilized almost as abundantly as sugar by
the adult plant. These observations were confirmed by Clark.? The
mycelium of Penicillium puberulum developed very slowly from
mycelium inoculation on 1.5 per cent alcohol. In the course of a few
weeks a delicate, thin, green growth spread over the surface of the
medium. Except for the green color it had the appearance of the
scum of lead oxid that forms on the surface of molten lead exposed to
the air.
The peptone cultures turned exceedingly dark. When the alkaline
medium was extracted with chloroform, the residue of the extract
consisted of a little oil which was for the greater part soluble in acid.
The acid solution gave a decided precipitate with Meyer’s reagent for
alkaloids. The small quantity available was not toxic to mice.
When the medium was acidified before extraction a little nontoxic
oil passed into the chloroform. By extracting with warm water no
penicillic acid was obtained. However, on long standing with ferric
chlorid the extract developed a faint rose color.
The purpose of the leucin and tyrosin cultures was to ascertain
whether Penicillium puberulum is able to deamidize amino acids as
yeast does. If this were the case amyl alcohol would have been
found in leucin cultures and tyrosol or tyrol in tyrosin cultures.*
Amyl alcohol was sought by the method of Beckmann,‘ and tyrol
and tyrosol by the method of Ehrlich.’ Raciborski ° first observed
differences in the manner in which different molds decompose tyrosin.
He found that P. glaweum grown on tyrosin agar produced a sub-
stance reducing silver. Since both leucin and tyrosin are destroyed
by P. puberulum, it is evident that either their decomposition differs
from that to which these amino acids are subjected by yeast, or else
amyl alcohol, tyrol, and tyrosol are merely intermediary products.
1 Duclaux, E. Surla nutritionintracellulaire. Annales del’Institut Pasteur, ann. 3, p. 97-112, 1889.
2Clark, J. F. On the toxic effect of deleterious agents on the germination and development of certain
filamentous fungi. Botanical Gazette, v. 28, p. 385, 1899.
8 After the completion of these experiments, Ehrlich and Jacobsen reported that Penicillium glaucum is
able to decompose amino acids to simpler compounds of lower molecular weights. Still more recently,
Herzog and Saladin published similar results. See Ehrlich, Felix, and Jacobsen, K. A., Uber die Umwand-
lung von Aminosiiuren in Oxysiiuren durch Schimmelpilze. Berichte der Deutschen Chemischen
Gesellschaft, Jahrg. 44, p. 888-897, 1911; Herzog, R. O., and Saladin, O., Uber das Verhalten einiger Pilze
gegen Aminosiiuren, Zeitschrift fiir Physiologische Chemie, Bd. 73, p. 302-307, 1911.
4 Beckmann, Ernst. Zur Bestimmung des Fuseldlgehaltes alkoholischer Flissigkeiten. Zeitschrift fir
Untersuchung der Nahrungs- und Genussmittel, Bd. 10, p. 143-152, 1905.
6 Ehrlich, Felix. Uber die Vergiihrung des Tyrosins zu p-oxyphenylithylalkohol (Tyrosol). Berichte
der Deutschen Chemischen Gesellschaft, Jahrg. 44, p. 139-146, 1911.
6 Raciborski, M. Uber die Assimilation der Stickstoffverbindungen durch Pilze. Bulletin Internatior
de l’Académie des Sciences de Cracovie. Classe des Sciences Mathématiques et Naturelles, ann. If
p. 767. 1907.
270
,
J
7
Vi
.
;
4
J
u
PENICILLIUM PUBERULUM. 37
That the latter is probably the case is indicated by the fact that
penicillic acid is formed abundantly only under conditions of imper-
fect aeration. Perhaps under these conditions tyrosin and leucin
would be less completely oxidized. It certainly is significant that
yeast which grows anaerobically produces tyrosol or g-oxyphenyl
alcohol from tyrosin, a substance which resembles penicillic acid in
bitter, toxic,! and certain chemical properties. Another observa-
tion of Raciborski? suggests a different explanation of the results.
He found that Aspergillus mger in a condition of carbon hunger
decomposes tyrosin in a different way than when it is plentifully
supplied with sugar. It is possible that since P. puberulum was
allowed to grow more than a month it was, during the latter period
of its growth, in a state of carbon starvation because all the sugar
had been consumed.
The amyl-alcohol cultures were designed to learn whether this
alcohol could be oxidized by P. puberulum. It was thought that
in this way some indication might be given showing whether
it was an intermediary product in the decomposition of leucin.
Unfortunately, as inspection of Table IV shows, amyl alcohol is so
poisonous that the question can not be definitely settled in this
way. The percentage tolerated is so small that the quantity can not
be determined with sufficient accuracy. It may, however, be stated
that there is no evidence that amyl alcohol is oxidized by P. puberu-
lum, since after several weeks’ growth it had not disappeared from
cultures containing as little as 0.05 per cent. In these cultures it
was separated by the method of Beckmann, the characteristic odor
being recognized in the final extract. These observations probably
have little bearing on the question of the intermediary formation of
amyl alcohol from leucin. As far as they indicate anything they are
not in favor of it.
In both leucin and tyrosin cultures an appreciable amount of vola-
tile acid was found, possibly formic acid, since the silver salt was
rapidly reduced. Because of this property the acid from the leucin
culture was not studied. That from the tyrosin culture formed a
erystalline barium salt. The quantity was too small for analysis.
The tyrosin culture presented a number of peculiarities. While
the musty odor of the growth on ordinary Raulin’s medium was per-
ceptible only when the mycelium was held close to the nose, that on
tyrosin had a typical and extremely musty odor which permeated the
room when the flask was opened. The odor of ordinary cultures be-
came more perceptible when they were distilled, since the odoriferous
1 Ehrlich, Felix. Uber die chemischen Vorgiinge des pflanzlichen Eiweissstoff wechsels und ihre Bedeu-
tung fiir die alkoholische Girung und andere pflanzenphysiologische Prozesse. Landwirthschaftliche
Jahrbiicher, Bd. 38, Erginzungsbd. 5, p. 306-307, 1909.
2 Raciborski, M. Op. cit., ann. 1906, p. 765. 1907.
270
38 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
principle seemed to accumulate in the distillate. Thom? observed —
that the production of odors varied greatly not merely from species to —
species, but also in some individual species according to the culture —
medium. It would be interesting to investigate whether in the ©
present case the production of the odorous principle was actually
dependent upon the presence of an aromatic compound or upon some
other condition. Such factors as these may be involved in the pro-
duction of the flavor of cheese.
The rate of growth was very much more rapid at first than in the
controls, in spite of the fact that little more than half the tyrosin was
consumed. Moreover, spore formation began before the fifth day
and was very abundant. In less than a week the entire surface of
the liquid was covered with mycelium uniformly green with spores.
It was more delicate and less stiff and woody than that of the controls.
It was smooth and even, and lay flat on the surface, whereas in the
controls it was convoluted and twisted so that some of it was pushed
below the surface, resulting in the formation of new mycelium above.
In the tyrosin the growth, after the surface had once been covered,
seemed less bulky, although the old mycelium was gradually over-
grown with new mycelium.
Certain of the facts here recorded have some theoretical interest.
This is particularly true of the peculiarities of growth of the tyrosin
culture and of the absence of easily detectable oxidizing enzyms in
al] the cultures tested.
The peculiarities of the tyrosin culture which are of interest in
this connection are the thinness of the mycelium and precocity of
the spore formation. Tyrosin contains the aromatic ring. Perhaps
a large amount of aromatic derivatives is required so that spore
formation can not take place until the organism has had time to
manufacture these aromatic compounds from the sugar and other
straight-chain carbon compounds offered. When an assimilable
aromatic compound is offered, this latent period is perhaps bridged
over. Certainly, on the tyrosin the spore formation is at least as
rapid as on corn-meal mush, which, in its proteins, contains an —
abundance of aromatic compounds. The importance of various —
amino acids for microbic growth has recently been brought out in
studies on the cultivation of the leprosy bacillus.?, Another explana-
tion may, however, be based on the assumption that spore formation —
is rapid in an exhausted or unfavorable medium. That this is
actually true is still an open question, though Tiraboschi* presents |
1 Thom, Charles. Cultural studies of species of Penicillium. U.S. Department of Agriculture, Bureau
of Animal Industry, Bulletin 118, p. 90, 1910. -
2 Duval, C. W. Cultivation of the leprosy bacillus from the human tissues, with special reference o-
the amino acids as culture media. Journal of Experimental Medicine, v. 13, p. 365-373, 1911. ‘
8 Tiraboschi, C. Studi sugli ifomiceti parassiti del granturco guasto. Atti del Terzo Con
Pellagrologico Italiano, Milano, 24-25-26 Septembre, 1906, p. 138. 1907.
270
PENICILLIUM PUBERULUM. 39
evidence for this view. If this hypothesis be accepted it would be
necessary to assume that tyrosin is an unfavorable source of nitro-
gen, either because of its chemical properties or because of its insolu-
bility. That its chemical properties are the cause seems hardly
likely, since leucin, which also contains amino nitrogen, gives such
an abundant growth, although Raciborski! states that tyrosin is a
much poorer source of nitrogen for Aspergillus niger than ammonia.
That its insolubility is the cause is quite possible. Tyrosin is so little
soluble that the amount in solution at any moment would be very
much less than in the unmodified Raulin’s medium, for most of the
tyrosin contained in the culture had crystallized out on the bottom
of the flask, at least 5 centimeters below the mycelium. Hence, it is
possible that the rapid sporification was a sign of nitrogen starvation,
which is the more likely, as the culture was very little handled or
agitated. Under these conditions diffusion is very slow indeed,
particularly for substances of great molecular weight like tyrosin.
That diffusion does not keep pace with consumption of material is
evident if any flask culture which has remained undisturbed for
some days be examined. If such a culture be held between the eye
and the light and gently rotated, convection currents can be seen
near the mycelium, showing that the specific gravity of the liquid in
immediate contact with the mycelium is very much less than that of
the deeper layer. It may therefore be that after the first few days
the mycelium was in relative nitrogen starvation. This hypothesis
would account for the rapid initial and slow subsequent growth.
A perusal of the literature on the metabolism of the fungi shows
that this question of the rate of diffusion has not received adequate
attention. If the organism is grown on a thick layer of culture fluid
the rate of diffusion of the dissolved substances is undoubtedly an
important factor. A substance of small molecular weight will diffuse
upward more rapidly to' replace that consumed than one of great
molecular weight. Thus the substance of small molecular weight
might appear to be a better food relatively than it really is. The
same logic applies to metabolic products. When these products are
substances like alcohol, of light specific gravity, they will accumulate
at the surface, particularly if the viscosity of the culture fluid is
great, owing to‘the presence of much sugar or peptone. If, then, the
alcohol concentration be determined for the whole fluid the values
obtained may be spurious. Actually the mycelium may have been
in contact with a much higher concentration of alcohol. This may
be one of the reasons why the crop of mycelium for a given amount of
sugar is said to be greater when the organism is grown on a thin
1 Raciborski, M. Uber die Assimilation der Stickstoffverbindungen durch Pilze. Bulletin Inter-
national de l’ Académie des Sciences de Cracovie. Classe des Sciences Mathématiques et Naturelles, ann.
1906, p. 764. 1907.
270
40 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
layer with great surface than when grown on a thick layer of culture —
fluid withsmaller surface.1. The largest yields were obtained in this way.
The absence of easily detectable oxidase is significant in connec-
tion with the discovery of Euler and Bolin that the oxidase of alfalfa
(Medicago sativa) is a mixture of the calcium salts of simple oxy-
acids.2 It is believed by some that many molds do not require
calcium.? It is tempting to imagine a connection between the
absence of easily detectable oxidase, the observation of Euler and
Bolin, and the absence of calcium.
GENERAL CONSIDERATIONS.
Since it has been definitely shown in the present paper that a
distinct species of Penicillium produces a substance of moderate
toxicity, the question very naturally arises, has it any pathological
significance? At present it can only be said that it is too early to
answer this question. All that can be done is to discuss the possi-
bility and to indicate further work to be done.
In acute intoxications, alleged to be due to molds, penicillie acid
alone can hardly be of significance. The lethal subcutaneous dose
for mice, as has been shown, is about 0.3 gram per kilogram of body
weight. Assuming the same susceptibility for the average man of
about 70 kilos, the dangerous dose would probably be about 21 grams.
Hence, an acute intoxication from penicillic acid would require that
an inconceivably great quantity of moldy food be consumed in a
single day. Even herbivorous animals could hardly eat enough
moldy fodder in a day to be acutely affected by penicillic acid. It is
quite out of the question that in the natural course of events penicillic
acid is likely to produce acute intoxication.
However, the results of this investigation of Penicillium puberulum
indicate the possibility of acute intoxication by moldy food. As
already stated, the different species of Penicillium differ radically in
their biochemical behavior. If there is so much difference in the
ordinary products of metabolism, it is altogether likely that a series
of toxic substances may be produced by different species. Some of —
these substances might very well be far more toxic than penicillic
acid. Indeed, Italian investigators have shown this contingency to —
be very probable. Gosio, Di Pietro, and Sturli have obtained from —
pure cultures of Penicillium toxic extracts far more poisonous than
1 Nikitinsky, J. Ueber die Beeinflussung der Entwickelung einiger Schimmelpilze durch ihre Stofl- _
wechselprodukte. Jahrbiicher fiir Wissenschaftliche Botanik, Bd. 40, p. 43, 1904. d
Raciborski, M. Op. cit., p. 733-734. 4
2 Euler, H.,and Bolin,I. Uber die chemische Zusammensetzung und die biologische Rolle einer —
Oxydase. Zeitschrift fiir Physikalische Chemie, Bd. 69, p. 187-202, 1909. a
3 Loew, Oscar. Uber die Giftwirkung von oxalsauren Salzen und die physiologische Funktion "a
Caleciums. Biochemische Zeitschrift, Bd. 38, p. 226-243, 1912. a
Robert, Mile. Influence du calcium sur le développement et la composition minérale de l’ Aspergil gy
niger. Comptes Rendus del’ Académie des Sciences [Paris], t. 153, p. 1175-1177, 1911. FA
270
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PENICILLIUM PUBERULUM. 4l
anything hitherto obtained in the new work herein recorded. Because
none of these investigators have isolated the toxic principle in a
state of purity their researches have not been given the serious con-
sideration that is their due. It will be the task of this laboratory to
extend these investigations to other species of Penicillium in the
hope that other toxic substances, perhaps more active than penicillic
acid, may be isolated.
In the matter of chronic intoxication the situation is quite different.
Continued use of moldy food containing penicillic acid might produce
symptoms. The quantity of badly spoiled corn-meal mush which a
man would consume at a single meal mighi contain as much as 0.1
to 0.5 gram of penicillic acid. As this acid has a toxicity of the same
order of magnitude as phenol, resorcin, or salicylic acid, and as such
substances are believed by many to be undesirable as food preserva-
tives, it seems reasonable to demand that great care be exercised in
eliminating moldy corn from the diet. Owing to the difficulty of
procuring material, it has not been possible to conduct long-continued
feeding experiments. Therefore it is impossible to say whether
penicillic acid has cumulative action. Should it prove to have such
action chronic intoxication might be brought about by comparatively
small doses. For this reason it is very desirable to learn the consti-
tution of penicillic acid in order to be able to make it synthetically.
This is the most promising way to obtain larger quantities of it.
While the finding of penicillic acid indicates that the relation of
moldy corn to pellagra’ deserves renewed attention, this discovery
does not materially strengthen the maize theory of the etiology of
pellagra. Penicillic acid itself is not sufficiently toxic. It is quite
possible that penicillic acid or a closely related substance may have
been responsible for the toxic effects following the administration of
‘‘nellagrozein,”’ the poison obtained from spoiled maize, with which,
according to the experiments of Lombroso,? the disease could be
produced artificially. ‘‘Pellagrozein”’ itself Lombroso did not regard
as anything but a mixture. He believed it contained two alkaloids,
which accounted for the toxic action. Neither alkaloid has ever
been obtained in a state of purity, so that it is impossible to form
any definite opinion about them. Indeed, other investigators have
not been able to find alkaloids at all in spoiled maize.? It is quite
1Cf. Marie, A., Pellagra, authorized translation from the French, by C. H. Lavinder and J. W. Bab-
cock, Columbia, S. C., 1910.
2 Lombroso, Cesare, and Erba, Carlo. Sulle sostanze stricniche e narcotiche del mais guasto. Reale
Istituto Lombardo, Rendiconti, s. 2, v. 9, p. 123-147, 1876.
8 Monselise, G. Ricerche chimico-tossicologiche intorno ad alcuni campioni di mais per la studio della
pellagra, Mondovi, 1881, 58 p. (Cited by Gosio. )
Selmi, Antonio. Delle alterazioni alle quali soggiace il granturco (Zea mais) e specialmente di quello
che ingenera la pellagra. Atti della R. Accademia dei Lincei, s. 3, Memorie della Classe di Scienze Fisi-
che, Matematiche e Naturali, v. 1, dispensa 2, p. 1099-1141, 1877.
Di Pietro, Melchiorre. Sui veleni dialcune muffe. Annalid’Igiene Sperimentale, v. 12 (n.s.), p.314-
365, 1902.
270
42 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
possible that these alkaloids were either normal constituents of maize —
or ptomainelike bases, as was pointed out by Pelloggio,’ for the inves-
tigators who found alkaloids seem to have often allowed the maize —
to spoil to an extreme degree. In one case the maize was actually
allowed to rot until it stank.? If the maize used contained either
penicillic acid or some similar substance the method of preparation
was such that these substances would have passed into the “ pellagro-
zein,’’ which is even less toxic than penicillic acid. The lethal sub-
cutaneous dose varied from 1.5 grams per kilogram for frogs to 7 to
10 grams for cats.* Hence, it is not impossible that ‘‘pellagrozein”’
contains substances of this type. Whatever evidence there is for
the relation between ‘ pellagrozein”’ and pellagra would apply equally
well to penicillic acid. The discussion of this question lies beyond
the scope of the present paper.
PENICILLIUM STOLONIFERUM.
In a former publication * it was stated that most of the samples of
spoiled American maize examined in this laboratory failed to give
Gosio’s® phenol test with ferric chlorid. In this respect American
spoiled maize seems to differ from that found in Italy, where the
ferric-chlorid reaction is regarded as a reliable test for the deterioration
of maize.®
Since the publication of the above-cited studies of the deterioration
of maize the test has been improved in this laboratory so that in its
new modification it is more delicate. The procedure as now con-
ducted consists in extracting 50 grams of ground corn or meal in a
stoppered flask, with sufficient chloroform to cover the mass. After
two hours the chloroform extract is filtered off and concentrated to a
bulk of 10 to 15 cubic centimeters. This concentrate is transferred
to a small separatory funnel or test tube and covered with 5 cubic
centimeters of water containing a trace of ferric chlorid. If sub-
stances like penicillic acid are present, the characteristic color
develops in the aqueous layer.
1 Pelloggio, Pietro. Materia reagente quale alcaloide, trovata nell’ estratto.del mais guasto preparato
dall’ Erba. Reale Istituto Lombardo, Rendiconti, s. 2, v. 9, p. 118-121, 1876.
? Lombroso, Cesare, and Erba, Carlo. Loc. cit.
Biffi, S. Sulla nota del prof. Cesare Lombroso: I veleni del mais e la pellagra. Reale Istituto
Lombardo di Scienze e Lettere, Rendiconti, s. 2, v. 9, p. 282-288, 1876. ;
8’ Lombroso Cesare. I veleni del mais e la loro applicazione all’igiene ed alla terapia. Rivista Clinica —
di Bologna, s. 2, ann. 7, p. 109-112, 1877.
—— and Erba, Carlo. Op. cit.
‘Black, O. F., and Alsberg, C. L. The determination of the deterioration of maize, with incidental —
reference to pellagra. U.S. Department of Agriculture, Bureau of Plant Industry, Bulletin 199, 1910.
* Gosio, B. Ricerche batteriolugiche e chimiche sulle alterazioni del mais, Rivista d’ Igiene e Sanita
Pubblica, ann. 7, p. 825-849, 869-888, 1896. 1
® Gosio, B. Alterazioni del granturco e loro profilassi. Italy, Direzione generale dell’ Agricoltura,
Annafl di Agricoltura, no. 261, 1909. ;
270
PENICILLIUM STOLONIFERUM. 43
When the tests are conducted in this way the number of samples
of obviously deteriorated maize showing the reaction is greater than
when the unmodified Gosio test is employed. Nevertheless, a
positive result seems to be less frequent in American maize than in
Italian maize. Moreover, the colors obtained with American spoiled
maize have always been found to be red or red brown, while in Italy
tests of spoiled corn are most commonly described as showing violet,
blue, purple, and greenish tints. None of these tints have been en-
countered in American maize in this laboratory.
Since this sharp difference apparently exists between American
and Italian deteriorated maize, it is desirable to compare samples of
Italian spoiled maize with American ones. Opportunity to make this
comparison was offered by Dr. C. H. Lavinder, of the Public Health
Service, who while on a visit to Italy very kindly secured samples
of condemned maize.
From one of these samples of maize Dr. E. F. Smith, of this Bureau,
isolated two species of Penicillium. One of these species was iden-
tified by Dr. Charles Thom, of the Storrs Agricultural Experiment
Station, as P. stoloniferum Thom.'
This organism when grown on Raulin’s medium gives the very
strong and characteristically violet ferric-chlorid reaction of Gosio.
It is certainly a remarkable fact that the first sample of spoiled
Italian corn examined gave the violet color described by Italian
authors, whereas no American sample has been found giving a similar
tint.
It was therefore decided to isolate, if possible, the substance
responsible for the ferric-chlorid reaction. For this purpose the or-
ganism from Italian spoiled corn was grown in “ Long Blake”’ bottles
on Czapek’s medium and on Raulin’s medium in the manner above
described. It was found that the organism grew more rapidly upon
Raulin’s medium. Therefore, for the preparation of material
Raulin’s medium only was used.
The substance responsible for the ferric-chlorid reaction was
isolated by the following procedure: The culture fluid and the myce-
lium were transferred to an evaporating dish and rendered weakly
alkaline with sodium carbonate. The contents of the dish were
then heated to boiling and filtered hot. The mycelium remaining
on the filter was thoroughly expressed. The mass was then again
extracted with water rendered weakly alkaline with sodium car-
bonate. The combined extracts were evaporated to a small bulk
over a free flame and filtered hot. To the clear filtrate a slight
excess of hydrochloric acid was ddded. An abundant precipitate
1Thom, Charles. Cultural studies of species of Penicillium. U.S. Department of Agriculture, Bureau
of Animal Industry, Bulletin 118, 1910.
270
44 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
was produced, which consisted of a mixture of needle clusters and
amorphous material. The precipitate was separated by filtration —
and washed with cold water. After drying spontaneously it was —
extracted with hot toluene and the hot extract filtered. Only the ©
crystalline portion of the precipitate dissolved. The amorphous
dark-brown material which remained on the filter was discarded,
for it did not give a color reaction with ferric chlorid. On cooling
and evaporating, the toluene extract spontaneously precipitated in
the form of needles, the material giving the ferric-chlorid reaction.
These needles, which were still slightly colored, were finally obtained
white either by decolorizing with boneblack in hot toluene solution
or by dissolving in alcohol and adding alcoholic potassium hydroxid
to form the potassium salt, which is insoluble in alcohol. This salt
was then washed free from color with alcohol. From the potassium
salt the free acid was recovered in the form of white needles by
dissolving the salt in water and precipitating with hydrochloric acid.
The substance thus obtained consists of white needles with a
melting point of 140° C., uncorrected. The name mycophenolie acid
is provisionally suggested for it. It is almost insoluble in water,
but freely soluble in alcohol, in ether, and in chloroform. It is some-
what less soluble in benzene, only moderately soluble in cold toluene,
and very soluble in hot toluene. With ferric chlorid it gives a violet
color in aqueous solution, though its solubility in water is not suffi-
cient to render the color intense. In alcoholic solution it gives a
bright-green color with ferric chlorid. It does not react with Millon’s
reagent. It does not give Lieberman’s reaction and could not be
diazotized. It does not reduce Fehling’s solution nor ammoniacal
silver nitrate. It is fairly resistant to sodium, ammonium, and
potassium hydrates and to hydrochloric, sulphuric, and acetic acids,
being unaffected by boiling in 10 per cent solutions of any of these
reagents. It does not contain water of crystallization. Its salts
of potassium and sodium are very soluble in water. The salt of
potassium is soluble in dilute alcohol, but insoluble in absolute
alcohol. The salt of sodium is soluble in absolute alcohol, but may
be precipitated in crystalline form by adding ether. The salt of
barium is only very slightly soluble in water and forms clusters of
minute needles. The copper, lead, and silver salts are amorphous
and insoluble in water. In characterization of the substance the facts —
collected in Table V were ascertained by analysis of the free acid, by
titration of the alcoholic solution of the free acid with n/10 sodium ~
hydroxid, and by the determination of the barium content of thes
salt on ignition in platinum with sulphuric acid. |
270
PENICILLIUM STOLONIFERUM. 45
TaBLeE V.—Analyses of mycophenolic acid.
Weight of
substance.
Gram. Gram. | Gram. | Per cent. | Per cent.| Gram. | Per cent. C.¢.
9/216. isk 0.5419 | 0.1315 | 63. 81 Cd TES Ream, Nay SPT
Db 5... .4770 | .1161| 63.64 TRS ERS SERENE AIRES co ty ce
NOTED BE Ga la PN CT ATS ee 12 SG a 0. 1256 WIG He io. t1e
AES SCENES SGD 8 EN SE EN CELE 11.53
FS C3 a [oy OR ol eae 63. 725 CAS Sy EA 2 pal Re SUR RSEE S| Speen ee Ae
Calculated for C,,H»0,: carbon, 63.74 per cent; hydrogen, 6.25 per cent.
RE ye carbon, 63.72 per cent; hydrogen, 6.30 per cent.
Calculated for Ba (C,,H,,O,): barium, 29.15 per cent.
SS a barium, 30.09 per cent.
A molecular weight determination by the elevation of the boiling
point in chloroform solution gave the results shown in Table VI.
TasBLeE VI.—Ebullioscopic determination of the molecular weight of mycophenolic acid.
Weight of Weight of Roileis Molecular
substance. solvent. point. weight.
Gram. Grams. Degree C.
et eee 30. 32 0. 065 308
Gor. a 30. 32 060 321
PA GTAGR Te Byer hc cieicoe | oust aereeee Ss 314.5
Molecular weight calculated for C,,H.,O,........-...------------- 320
Molecular weight found from titration......... svi ALU. DOSER
Molecular weight found from barium eid mp at ucmetiaz hess 328
Molecular weight found from boiling-point elevation.........-..-- 314. 5
The formula C,,H,,O, may therefore be assigned to mycophenolic
acid. It does not readily decompose carbonates at ordinary tempera-
tures. It is apparently a dibasic acid, or, at any rate, combines
with two atoms of a monovalent base. Whether the base combines
entirely with carboxyl groups or with phenol groups has not been
determined.
The acid seems to form two series of salts. Presumptive evidence
on this point was obtained by the following experiments: Two deci-
grams of free acid were suspended in water and one equivalent of
potassium hydroxid added. Unfortunately, this quantity was not
sufficient to dissolve the substances completely, so that a slightly
greater quantity of the alkali had to be used. This solution was then
treated with one equivalent of barium chlorid. On standing in the
desiccator a crystalline barium salt formed. This salt was evidently
different from the normal barium salt, which is so insoluble that it
precipitates at once. It was also of different appearance under the
micwscope, consisting of a fewsmall needles in clusters, which appar-
" 270
46 CONTRIBUTIONS TO THE STUDY OF MAIZE DETERIORATION.
ently were the normal salt, and more abundant larger single needles, —
apparently the acid salt. The presence of the normal salt in small —
quantities under the conditions of the experiment was probably due
to the fact that an excess of alkali had to be used in dissolving the ©
substances. The barium content of this preparation was deter-
mined, 0.207 gram yielding 0.0692 gram of BaSQ,, equivalent to a
barium content of 20.2 per cent.
Calculated for Ba.( Op Mb asec tempi - |