lok 53 |.$3 1915 Copy 1 » | Laboratory Outlines for General Botany FOR THE ELEMENTARY STUDY OF PLANT STRUCTURES AND FUNCTIONS FROM THE STANDPOINT OF EVOLUTION FOURTH EDITION BY JOHN H. SCHAFFNER PROFESSOR OF BOTANY, AND HEAD OF THE DEPARTMENT OF BOTANY Ono STATE UNIVERSITY COLUMBUS, OHIO PUBLISHED BY THE AUTHOR 1915 LABORATORY OUTLINES RI 86 GENERAL BOTANY For the Elementary Study of Plant Structures and Functions from the Standpoint of Evolution FOURTH OREVISED: EDITION BY JOHN H’¥ SCHAFFNER Professor of Botany and Head of the Department of Botany Ohio State University COLUMBUS, OHIO PUBLISHED BY THE AUTHOR iQI5 COPYRIGHT, rors BY JOHN H. SCHAFFNER MLW 3 & F'IS. PREFACE. The series of outlines presented in the following pages was first published in the Journal of Applied Microscopy. The outlines as here presented have undergone various additions and alterations in order to bring them up to date and to give a more perfect view of the plant kingdom as a whole. Among other things, an attempt has been made, so far as possible, to use a reasonable terminology. It is presumed that the course can be covered practically as given in one college year with three laboratory periods of two hours each a week. The course is intended for the freshman or sophomore year. The student should have a fair knowledge of language, mathematics and drawing as well as the foundations of chemistry, since the pursuit of the biological sciences calls for considerable independent effort, skill of manipulation, and ability to reproduce and describe what is seen. In case a briefer course is necessary a considerable number of types or parts of outlines can be omitted without seriously breaking the continuity of the subject. According to the author’s views any thoro course should include at least the follow- ing, either complete or in part: eye Vel Ie Woes CONT oa XOX COC Ge OX WOEEX VT EX... DOOGMaX XXIV XXXVIEXXXIX. G), XL-XEILE (e);, (ce), XLIV-XLVII., Pane ii IVE xX LXE XV. (a) LXVIELXVINT., LXX:-LXXIIT.. (a), ema = Cd). Cy. (gy eV I EX I aX XX EX X XT, EXXXV.- meee XC XCVL-Cle Cli, Civ, CVILKCX. -OXt Some of the types which cannot be given in the laboratory period may be used for class demonstrations. The appendix on microtechnique will be found useful where it is possible to have students prepare some of their own slides. The methods given are for the most part such as have been thoroly tested in the class room by the author himself. In this fourth edition a number of changes have been made in order to include recent advances in botany and related subjects. eels Se (3) Eye-piece Draw-tube Coarse adiust- ment: Ra k and pinion Tube Fine adjustment: Continuous Safety Micrometer Movement Dustproof triple nose. piece with objectives Abbe condenser Iris diaphragm Mirror Base PLATE [. COMPOUND Microscope. BAuscH AND Lomp Microscore FF S8, INTRODUCTORY SUGGESTIONS. The following outlines are designed for those who have access to little appa- ratus outside of a good microscope. The course will] probably be more than sufficient for the time usually alloted in most of our colleges and universities. Whatever may be the opinion in regard to the elementary course of botany, it is the writer’s belief that the general college or university course should be largely carried on with the use of the compound microscope; and should cover, in a general way, the whole plant kingdom, so that the idea of the evolution of plants and their natural relationships will be made prominent. The student should have a general grasp of the plant kingdom as a whole, and to accomplish such a result a large number of forms must be studied. Along with this general idea, a considerable knowledge of morphology and physiology may be acquired, since the study should have to do largely with living material. The course should cover a year with at least two laboratory periods of two hours each, a lecture, and a quiz with assignments from a suitable text-book. After such a course the student is well fitted to take up the various departments of advanced work. He will have acquired a sufficient knowledge of biology to carry on intelligently, whatever special studies the may later choose to pursue, as, anatomy, histology, cytology, physiology, ecology, taxonomy, genetics, or advanced work in special groups. It is often supposed that to accomplish good work it is necessary to have on hand an expensive equipment and all the facilities which our leading universities afford. There is, however, a large amount of work that may be done by those who do not have such an equipment, and substantial progress may be made in the general facts of the science with little besides what is indicated below. The student should have the following equipment: l. A good text-book of botany for general reading. 2. A compound microscope, like the Bausch and Lomb FFS®, having a double nose-piece with 16 and 4 mm objectives, and 5x and 12.5x eye-pleces ; the Spencer microscope No. 44 with 16 and 4 objectives, and 6x and 10x eye-pieces; or the Leitz microscope Stand II. U. with objectives 3 and 7 and eye-pieces II. and IV. The same stands with complete substage and triple nose-piece are preferable if one can afford to pay the difference in price. With the Bausch and Lomb stand the third objective may be the 48 mm, and this may be used instead of a dis- secting microscope. A number of slides and cover-glasses. 4. A good hand-lens or a dissecting microscope. In case the microscope has a triple nose-piece it should be fitted with a 48 mm objective. 9. A good note-book with note paper and smooth drawing paper, and also some bristol board drawing paper for the finer drawings. (See “The Laboratory Note Book” in appendix. ) 6. Loose writing paper for making temporary records and calculations. 7. Two good lead pencils a No Sit and a (Nos 6H. Tt is also desirable to have a bottle of India ink or Higgins’ eternal ink and crow-quill or other suitable drawing pen so that the drawings may be finished in ink. ee (5) 6 LABORATORY OUTLINES FOR GENERAL BORA 8. The following instruments are necessary: a. A pair of forceps. b. Several medicine droppers. c. Some needles set in wooden or bone handles. d. -A scalpel. e) -N razor. f. Dishes, watch glasses, butter dishes, and bottles of various sizes. g. Plenty of clean cotton rags and some paper blotters. 9. The following simple reagents will be needed on the table: A small bottle of 50 per cent. aqueous solution of glycerin. A bottle of distilled or pure, boiled water. Iodin solution. Salt solution, saturated aqueous. A bottle of ninety-five per cent. alcohol. Sao oe If a greenhouse is not near, a window garden and aquarium become indis- pensable. Water plants kept in glass jars .with some small water animals, as water snails and water beetles, will usually grow with little or no attention. In most cases the jars should be covered. Many of the specimens may be preserved in various preserving fluids, and some may be dried. These will be found very convenient in case fresh material cannot be obtaained when desired. Microscopic plants may be preserved in water, in homeopathic vials, provided a drop of carbolic acid is added ot each bottle of material. Plants like mosses, liveworts, fleshy fungi, stems, roots, rhizomes, etc., may be preserved in 70 per cent. alcohol. The ordinary filamentous alge are usually well preserved in copper salt solution. (See appendix.) Myxomycetae in the fruiting stage, woody fungi, lichens, some liverworts and many other plants may be kept in a dry condition in ordinary paper boxes. Useful pamphlets on the use and care of the microscope are furnished by the Bausch and Lomb Optical Co., of Rochester, N. Y., and the Spencer Lens Co., of Buffalo, New York. The following suggestions are offered especially for the benefit of laboratory students, altho most of the directions will also be useful to the amateur micro- scopist working at home: The microscope must always be handled below the stage and never lifted by any part above the stage (unless one has an instrument of the new type with a rigid arm), otherwise the fine adjustment may be injured. The microscope is a very delicate instrument. It must not be inclined for general work, as temporary mounts will not stay in the field unless the stage is horizontal. While working, the observer should keep the side of the microscope with the coarse and fine adjustments toward him. The microscope is not to be moved about to obtain the light. This can be obtained from almost any direction by adjusting the mirror properly. Great care must be taken so as not to run the objective down into the diaphragm or onto the cover-glass and slide. The lenses of the microscope must not be touched with the fingers. They must be wiped only with a very clean, soft, cotton cloth or with lens paper.. They must be kept scrupulously clean. The student should learn the different combinations of low and higb powers immediately and how to change from one to the other without difficulty. The wiping rags should always be clean, and the slides and cover-glasses must be kept scrupulously clean. The student should learn at the beginning how to clean the cover-glasses without breaking them To do this, take the cover- LABORATORY OUTLINES FOR GENERAL BOTANY. Fi glass, moistened in water or alcohol, in the rag between the thumb and forefinger and hold it at the edges between the thumb and forefinger of the other hand. In making a mount air bubbles are to be avoided. To accomplish this, after the object has been placed on the slide and covered with a drop of water, hold the cover-glass at the edges between the thumb and forefinger and bring it down obliquely onto a needle held in the other hand, and then withdraw the needle gradually. The cover-glass will then settle down on the object sur- rounded by water. No water or other reagent must be on top of the cover-glass. If too much water has been put on the slide it may be removed with blotting paper. If the study of a good specimen cannot be finished in the given time, it may be preserved for a number of days by running a little fifty per cent. glycerin under the cover-glass. Reagents cost money, and are not to be poured out like water. The same is true of the material for study. This is often difficult to obtain and should be used with economy, and all good surplus material returned to the receptacle from which it was obtained. All objects studied are to be carefully figured and described. The drawings may be outlined with the 3H pencil and then finished with the 6H. If time is at hand, the drawing may be finished in India ink with a fine drawing pen. Learn how to keep the pencils sharpened to a fine point. After sharpening with the knife rub the point smooth on a piece of paper. The drawings are to be placed only on the front side of the drawing paper. The notes may be written continuously on both sides of the note paper, but are always to be taken down in ink. The plates containing the drawings should be numbered in Roman figures at the top, and the name of the plant or object written at the bottom. The separate drawings on the plate may be numbered in Arabic figures, and a proper necond of them is to be kept in the notes. Vhe notes on each plant may be numbered the same as the plate containing the drawings to illustrate it. The drawings should not be crowded, and the number should always be written below. The diameter of the field (the white disk visible when looking into the micro- scope) is usually about eight inches (two decimeters) when projected onto the table. Learn to do this by looking with one eye on the table beside the micro- scope and with the other into the tube. In this way the magnified image may be directly measured. The actual diameter of the area covered can easily be determine for the low powers by examining a millimeter rule. Learn to keep both eyes open when taking only ordinary observations in the microscope. Be sure to use both eyes, else one will be trained for more acute vision than the other. Make the drawings -of small objects of the right proportion, and the actual size magnified. The larger ones’ may have to be reduced to bring them onto the paper. If the object has a definite relation to environment do not draw it upside down. It must also be remembered that motions are magnified as well as the objects themselves. Absolute regard for the truth is the first requirement in scientific drawings and descriptions, and the qualities required for good work are accutacy, cleanli- ness, patience, skill, pesistency, good judgment, and logical ways of thinking. The drawing should be exact in all details; the sketches may be more or less diagrammatic. The notes should be written in the best English at the command of the student. The facts should be stated in concise but complete declarative sentences, without rhetorical ornamentation. The observations must always be recorded at the time when they are taken. One’s memory should not be trusted very much in recording scientific facts. g LABORATORY OUTLINES FOR CENERAL VS Oir inne Finally, it must be remembered that one of the first things to be accom- plished is to educate the hand for delicate manipulations. And it is also well to keep in mind that scalpels and razors are not intended for sharpening lead pen- cils or cutting the table, that oculars, and objectives are never to be dropped, that stoppers should not be laid down on the bare table, that books and note-books are not to be soiled by the wet and dirty fingers, that bottles and tumblers of water are not to be overturned, and that one should understand the objects studied before attempting to draw or describe them. ANTHOPHYTA, oj porn vg CYCADOPHY TA ~ » PTENOPHYTA “6 CALAMOPHY TA ean Aoisel BRYOPHY TA GONIDIOPHYTA fie HYTA A GHAROPHYTA. So » SCHIZOPHY TA Sea Eton ARCHEOPHYIA PLATE IJ. DIAGRAM oF THE PLANT PHYLA. PRELIMINARY STUDY OF THE LIVING CELL. J. Philotria canadénsis (Mx.). Waterweed. This is a very common plant growing submerged in ponds, creeks, etc. It will grow well for a long time if simply pulled up and placed in a covered glass fer. 1. Carefully pull off a few young leaves and mount on a slide with a drop of water and a coverglass. Examine under the dissecting microscope. Sketch the entire leaf under low power of the compound microscope. Make the drawing about five inches long. Describe the shape, margin, color, midrib. Are there any other veins? 2. The leaf is composed of cells. How many across the leaf? How many lenthwise? Is the leaf more than one cell in thickness? About how many cells on the upper surface? 3. Cut cross sections with the razor by holding some leaves between pieces or strips of common carrot either fresh or preserved in alcohol. How many cells in thickness, on the average? 4. Suppose the leaf averages three cells in thickness, about how many cells im the entire leaf? 5. Under high power, draw several adjoining cells, carefully showing details. (Draw the walls as represented in Fig. 1). What is the general shape of the cells? The contents of a cell are protoplasm and sap or water. There is usually some dead food material present. Ss ee iene, Jie == Caren, WONS Oi IP EnicO mien. 6. Draw a cell showing the nucleus. Notice that the protoplasm is made up of cytoplasm, nucleus, and chloroplasts. Where is the green coloring matter? What is the color of the rest of the leaf? The green coloring matter is chlor- ophyll. What is its use? Estimate the number of chloroplasts in a single cell. How many would there be in the entire leaf? How does a green plant get its food? (9) 10 LABORATORY OUTLINES FOR GENERAL BO mye 7. Movement of protoplasm. Describe the motion. Do not be satisfied until the rotation is very striking. The room and water should not be too cold. Does the protoplasm rotate in the same direction in all of the cells? How many seconds does it take for a chloroplast to make the round? Does the nucleus move in the cell? The active agent in the movement is the cytoplasm. The cytoplasm does not move from one cell to another. 8. A cell is a small mass of protoplasm, in typical plants usually differen- tiated into cytoplasm, nucleus and plastids, and surrounded by a cellulose wall. The cell is the unit of plant structure. In some of the lower plants no nucleus has been discovered, and in many plants the plastids are absent. 9. Treat a fresh leaf with alcohol. Does the protoplasm still move? What effect does the alcohol have on the chlorophyll? Treat a fresh specimen with salt solution. What takes placer Explain the cause. Ask fom anwexplanauon or study the subject of plasmolysis in a text-book. These cells have a vacuole (water chamber) inside of the protoplasm and are normally in a turgid con- dition. Treat the specimen in alcohol with iodin solution. Notice the nucleus and nucleolus. Notice the large starch grains stained dark blue inside of the chloroplasts. 10. Ecological note. Does this leaf have stomata? How is it adapted to its environment? II. Allium cépa L. Common Onion. 1. Pull off the inner and the outer epidermis from a living scale of an onion. Mount in water. Compare the cells of the two specimens under. low power as: to shape, size, and contents. Notice the wall lined with cytoplasm; also the nuclei. Draw a number of adjoining cells from the inner epidermis. Notice the absence of chloroplasts. 2. Under high power, draw a single cell showing the wall, cytoplasm, and nucleus. 3. Study the movement (streaming) of the cytoplasm. This can usually be seen best at the ends of the cells. Notice the fine strands of cytoplasm stretching across the cell or across the corners of the cell thru the large central vacuole. Make a diagram of a cell showing the position of these streams, and indicate the direction of the flow by means of arrows. 4. Treat with a drop of iodin solution after killing the cells in alchool. Make a careful drawing of the nucleus under high power showing the nucleoli. What is the normal number of nucleoli for each nucleus in these cells? Is the number constant? Are there any starch grains present stained blue by the iodin?’ 5. Why do the scales of the bulb not have chlorophyll? III. Tradescantia sp. Spiderwort. The flowers of almost any of the wild or cultivated species of spiderwort will be found suitable. Rhdeo discolar Hance, easily grown in greenhouses and window gardens, will also do very well. It blooms almost continuously. 1. Study the stamen hairs. ‘With a scalpel cut off some of the stamen fila- ments containing the young hairs. Mount in water. Be careful to get the hairs wet, but do not injure them. Under low power, notice that the hair is made up» of a chain of cells. Draw. LABORALRORY: OUTLINES FOR GENERAL BOTANY. 11 2. Study a single cell under high power. Observe the position of the nucleus; the cytoplasm, filled with small granules, lining the cell wall; and the large vacuole filled with water thru which granular strands of cytoplasm stretch. 3. Study carefully the streaming motion of the cytoplasm. Are the streams constant or can you see changes going on in their position? Do some of the strands disappear entirely? Watch the position of the nucleus for some time and describe its motion. Select one that is suspended in the central part of the cell. Make a large, careful sketch of a cell showing the streaming to good advantage. Plot all the moving streams visible by focusing up and down, and indicate by means of arrows the direction of the movement. SERIES I— THALLOPHYTA. IV. Pleurocéccus vulgaris (Menegh.). Phylum, Gonidiophyta. Class. Pleurococcee. Order, Pleurococcales. Family, Pleurococcacee. This is a unicellular green alga which very commonly forms a green, pow- dery layer on the bark on the north side of trees, on fences, rocks, etc., and is available at any time of the year. 1. Scrape off some of the green powder from a piece of moist bark and mount in water. Pick out one of the largest single plants and draw under high power, showing the thick cellulose wall and the chloroplasts. 2. Notice that the cells (individuals) ‘have a tendency to hang together for some time after division. Study and draw aggregates or colonies of two, three, four, and eight cells still united. In how many directions do the cells divide? Describe the color, shape, and habitat of the plant. How does it get its food? Notice that it must be exposed to long periods of drouth. Do not forget to look for this plant (and as far as possible all others studied) in its usual habitat out of doors. . he 520 : Os OB a OTe Ne ee Vi - yi b eae Fic. 2.— Lire CYcLe or PLEUROCOGCUS. 3. Its life cycle may be stated as follows: increase in size of the cell, division of the cell into two, separation of the daughter cells. Taking no account of the fact that the cells hang together for some time after division, make a diagram in the notes illustrating this as indicated in Fig. 2, a. 4. Make a diagram showing the ancestors of one individual for ten gen- erations. See Fig. 2, b. 5. Make a diagram showing the descendants of one individual for ten gen- eraitons . Use an entire page of note paper. See Fig. 2, c. 6. Note. All plants and animals, whether high or low (except cenocytic forms) are single cells in the first stage of their life. Therefore, in the higher forms, the egg or spore also passes thru the two, four, etc., celled stages, and in these first stages the cells may also represent a loose aggregate or colony, since in many cases, if the cells are separated from each other by artificial means, two (12) LABOrAnOiK OU PEINES (Ok GENERAL BOTANY. 13 or more plants or animals may be obtained from the egg, which would otherwise have produced only one individual. Pleurococcus shows how it is possible for a plant to pass from a unicellular condition to a colony, and from the condition of a colony to a multicellular plant. By what means could this be accomplished? The mechanical reason for the division of one of these cells may be dependent on the following facts: All food and waste material must pass thru the wall. Now the surface of a sphere is equal to D? and the volume is equal to ¢ 7 D’, therefore, as the sphere increases in size, the surface continues to become less in proportion to the volume. How could a cell increase indefinitely in size and still keep the surface and volume in about the same ratio? What disadvantage or limit would there be to such a process? ‘These plants have potential immor- tality, i. e., they do not grow old and die, except by accident. Natural death of an organism appears to be an acquired character. This plant, with a number of others to follow, is unicellular and without sexuality. It belongs to the lowest sub-kingdom of plants, which for convenience may be called the Protophyta. V. Merismopédia sp. Phylum, Schizophyta. Class, Cyanophycee. Order, Chroococcales. Family, Chroococcacee. This organism can usually be found in the sediment of creeks, ponds, or lakes, especially in shady places where there is some decaying vegetable matter. 1. Mount some of the sediment and examine under high power. Look for minute, blue-green, more or less rectangular plates of cells. Find colonies of various sizes, select a perfect one and draw, showing the arrangement of the cells. 2. In how many directions does cell division take place? How does the colony break up into smaller pieces? Such a flat layer of cells is called a super- ficial aggregate. Neither plastids nor nuclei are visible in these cells. The nuclei are very small. The bluish color is due to the presence of a peculiar coloring matter, phycocyan, in addition to the chlorophyll. Notice the gelatinous nature of the cell wall. Write a careful description of the plant. VI. Filamentous Blue-green Algae. (a) Lyngbya sp. Class, Cyano- phycee. Order, Oscillatoriales. Family, Oscillatoriacee. The species known as Lyngbya wollei Farl., which produces large brownish- black masses in rivers and ponds, or any other large species, may be used. A large species, appearing like a brown or black slimy layer, quite common in greenhouses and other moist situations, is also very good for study. This form can be kept indefinitely in a moist jar of earth. 1. Mount a small mass of the slimy material in water, and study under low power. Draw several of the greenish-brown threads or filaments showing how they are interwoven. Notice the disk-like cells which make up the filament. De- scribe the general character of these plants. 2. Under high power study a single filament. Draw part of a filament, show- ing the end cell. Why is the end cell more or less hemispherical and the others disk-shaped? Notice the dark granules. Where are they situated? Notice the thick sheath surrounding the cells. Draw a single cell, showing details as accurately as possible. 3. In how many directions do the cells divide? Where and how does cell division take place? A filament like this is called a linear aggregate. 4 14 LABORATORY OUTLINES FOR GENERAL BOTANY. 4. Reproduction. In old filaments look for the development of hormogones — short pieces of a number of cells broken loose inside of the shéath. Draw and describe. How do the hormogones escape from the sheath? (b) Oscillatoria sp. Family, Oscillatoriacee. Any of the minute, bluish-green forms which produce slimy, membranous layers in ponds, rivers and creeks may be used. They may be kept for an in- definite time in a covered glass jar of water. 1. Mount a small flake in water, study under high power, and draw several of the slender filaments. There is no definite sheath present. Describe the color, shape of cells, and cell contents so far as they can be seen. Are the two ends alike? Compare as to size, etc., with Lyngbya. Draw a single cell. 2. Study the reproduction. Compare with the method of reproduction in Lyngbya. 3. Make a careful study of the movement of the filaments. To get good results the plants should first be placed for some time in direct sunlight and the water should not be cold. Describe the movement. Why can these plants move more actively than the Lyngbyas? (c) Nodstoc commune Vauch. Class,Cyanophycee. Order, Nostocales. Family, Nostocacee. This plant is common on damp ground in meadows, pastures, hillsides, etc. After a rain it appears as dark green gelatinous wrinkled or lobed masses. It may be kept for an indefinite period and will be in good condition after soaking in water. tee 1. Describe the colony, noting its size, shape, and color. Draw. 2. Mount and under low power note the general arrangement of the fila- ments. Look for the limits of the thick gelatinous wall in favorable plants. 3. Under high power note the two kinds of cells composing the filament, ordinary cells and heterocysts. Why are the filaments so crooked? Draw a fila- ment showing both kinds of cells. VII. Beggiatéa alba (Vauch.). Phylum, Schizophyta. Class, Schizo- mycete. Order. Desmobacteriales (Filamentous Bacteria). Family, Beggiatoacee. These plants are usually very abundant in sulfur springs and in shady places in ponds and stagnant water where decaying vegetable matter is present. Beg- giatoa may be kept for years in a covered glass jar filled with water, provided there is a layer of decaying vegetable se\iment in the bottom. 1. With a medicine dropper take up some of the black sediment containing Beggiatoa, mount, and examine under high power. Study the slender, more or less hyaline filaments, and draw one carefully. Draw a single cell showing the large sulfur granules. No chlorophyll is present. Describe the plant in general. 2. Study and describe the movement. Do the sulfur granules move in the cell? How many seconds does it take for the tip of a filament to travel from one side of the field to the other? 3. How does this plant obtain its food, and upon what does it live? How different in this respect from Pleurococcus? To what physiological group does Beggiatoa belong; holophytes, saprophytes, or parasites? Note— These plants are intermediate between the blue-green alge and the bacteria. What relation is there between th lack of chlorophyll and th saprophytic habit ? as i EABOCAROK YT OU LDEINES FOR GENERAL BOTANY. 15 While Bacterta: (Class, Schizomycete. * Order, Bacteriales. There are three common families of bacteria: Coccacee, Spherical Bacteria, containing the genus Micrococcus and others. Bacillacee, Rod Bacteria, containing the genus Bacillus and others. Spirillacee, Spiral Bacteria, containing the genus Spirillum and others. To obtain Bacilli, make a hay infusion by boiling ordinary dry hay for 15 minutes. Keep in a sterilized covered dish for several days. Also boil some beans, and after exposing the broth to the air until cool, cover and set aside for two or three days. Species of Spirillum may be obtained from sewer watr, or by letting water plants decay in a jar of water. Micrococci are common in the air, and may be obtained on boiled potatoes, gelatin, moist bread, etc., by letting these culture media remain exposed for a short time and then covering them to keep in the moisture. The bacteria will soon begin to appear in yellow, pink, purple, or red patches. 1. Mount some of the hay infusion and examine under high power. Notice the minute free-swimming hay Bacteria, and draw several individuals. Draw several still hanging together in a filament after division. Describe the shape, color, and movement. Distinguish between the true locomotion of the Bacillus and the Brownian movement of the foreign particles present in the mount. 2. Draw two individuals with spores. The movement is produced by means of flagella or cilia. 3. Mount some of the bean broth and notice the putrid odor. Study the Bacillus present. Estimate the number of bacteria present in the field of the microscope. Counting the number across the diameter of the field and squaring will give approximate results. 4. Suppose you had one bacterium to begin with, and that it and its descend- ants divided once every hour how many bacteria would there be at the end of each hour for 48 hours? . 5. Mount some material containing Micrococcus. Draw several individuals and describe. 6. Mount and study some bacteria in the zodgleea stage (bacteria in gela- tinous masses.) Draw and describe. 7. Mount some water containing Spirillum. Study the peculiar movement. Draw several individuals and describe. Represent the cell as a real spiral and not simply as a wavy line. 8. Root-tubercle Bacteria. Collect fresh roots of white clover (Trifolium repens L.) or alfalfa (Medicago sativa L.). Sketch a rootlet showing numerous tubercles under dissecting microscope. Crush a large and a small tubercle on the slide, mount in water, and study the bacteria present. Draw a number of individuals, showing the following forms: irregular rods, club-shaped, T-shaped, Y-shaped, and X-shaped. Treat with iodin solution and note the color reaction of the starch and of the bacteria. This symbiosis is a case of mutualism. 9. Mount some hay Bacilli and some Parameecia tagether, and compare them as to size. The Bacillus and the Paramcecium are both single cells. About how much greater in volume is the Paramcecium than the Bacillus? In order to get fairly accurate results, find how many times wider, thicker, and longer the one is than the other. This can be done by projecting the organisms onto the table and measuring them with a millimeter rule. How near would the comparison hold with that of a mouse and an elephant? 16 LABORATORY OUTLINES FOR GENERAL: BOTANY. 10. NoreE—To obtain Parameecia, let a mass of Spirogyra or other water plants decay in a jar of water exposed to the air. The Paracecium is one of the most highly developed and specialized animals belonging to the Protozoa. IX. Slime Molds. Phylum, Myxophyta. Class, Myxomycete. Order, Myxogastrales. The Myxomycetes are a group of organisms which approach very near to the animal kingdom, forming one of the several transition groups from the lower plants to the lower animals. They have developed a very complex life history, altho they are very simple plants. They usually grow on decaying logs and stumps, and may be collected in summer and autumn and kept in the encysted or resting stage for an indefinite length of time, and studied when conveniet. (a) Lycogala epidéndrum (Buxb.). Family, Lycogalacez. 1. Make a sketch showing the naked eye characters of individuals in the resting stage (zthalia), and how they are situated on the wood. Describe. 2. Moisten some of the downy material (capillitium) and a piece of the outer enveloping layer (peridium) in alcohol; and mount in water. Examine under high power. Is there any cell structure in the capillitium or peridium? Draw a part of the capillitium, showing the peculiar markings. 3. Draw a few of the individuals (scattered thru the capillitium) in the rest- ing stage (spores), and describe. (b) Hemitrichia clavata (Pers.). Family, Trichiacee. 1. Mount one of the sporangia and sketch under low power, showing the stalk of the sporangium, the broken peridium, and the mass of capillitium threads. Describe shape, color, etc. 2. Under high power draw some of the capillitium threads, showing all details carefully. 3. Draw some of the individuals in the spore stage. (c) Stemonitis fisca (Roth.). Family, Stemonitacee. 1. Mount and draw one of the plume-like sporangia under the dissenting microscope, showing the hypothallus, stalk, columella, and capillitium. 2. Under high power, draw part of the capillitium, showing how it is attached to the columella. 3. Draw some of the spores. (d) Plasmodium. 1. .Examine under the dissecting microscope, and describe the plasmodium or a myxomycete in the moist living condition. This can usually be found on decaying logs during the spring, summer, and autumn. If living material is not at hand, examine pieces of plasmodium preserved in alcohol. 2. The flagellate stage of many species of myxomycetes may be obtained by simply making hanging drop cultures with water, or water in which decaying wood has been soaking. Fresh spores of Lycogala will germinate in a day or two, and the preparation can be examined from time to time under the high power. LABORATORY OUTLINES FOR GENERAL. BOTANY. 17 (c) Amoéba sp. Protozoa. Class, Rhizopoda. Order, Amcebida. Fam- ily, Ameebide. If the student has not studied the Amceba in a general course in zoology, it should be taken up at this point, since the amoeboid form probably represents one of the most primitive type of cells with which we have to deal. Amoebas can gen- erally be found in the ooze at the bottom of ponds and creeks. To obtain Amcebas in large quantities, pack a glass jar rather tightly with Ceratophyllum or with pond lily leaves, and cover with water. The dish should be covered up. After a week or two, when the plants begin to decay, Amoebas will usually be abundant. 1. Scrape off some of the sediment from the Ceratophyllum leaves and mount in water together with some of the brown scum present at this time in the jar. Under high power search for transparent, naked cells of irregular shape, which are slowly changing in outline by thrusting out pseudopedia. Sketch the outline of an individual six times successively, at intervals of ten seconds. 2. Describe the amceboid movement of the animal, and the formation of the pseudcopodia. 3. Make a careful diagrammatic drawing of a large Amoeba, showing the outer limiting layer (ectoplasm), the inner more fluid granular part (endoplasm), the nucleus (if distinguishable), the contractile vacuole, and the various ingested foreign bodies, as diatoms, desmids, etc. 4. Note.—In the form following, a return will be made to a typical plant related to Pleurococcus. X. Scenedésmus quadricatda (Turp.). Phylum, Gonidiophyta. Class, Pleurococcee. Order, Pleurococcales. Family, Scenedesmacee. Scenedesmus is very widely distributed, and may be found in the sediment in the bottom of ponds, creeks, etc., along with diatoms and other microscopic plants at any season. It usually consist of a colony of four, more or less spindle- shaped, green cells. The two outer cells have four slender, pointed, prong-like projections extending diagonally outward, one at each corner of the colony. 1. Mount some of the sediment containing Scenedesmus, and examine under high power. Draw and describe. 2. Compare a number of colonies as to size, shape of cells, and appearance of the projections. 3. In the sediment with Scenedesmus a simpler and much smaller green laga, Ankistrodesmus falcatus (Corda), belonging to the same family will probably be present. This consists of very slender, bent or doubly curved cells either separate or in masses. If present draw and describe. 4. Notre.— The thallophytes following are more highly developed forms and possess some type of sexuality or are forms supposed to be descendants from ancestors having a sexual process. They may be called the sub-knigdom Nema- tophyta. bez Wii WAN TP ASHLe CHIL WALLS -OR WITH: COMPLICATED CHROMATOPHORES, XI. Diatoms. Phylum, Zygophyta. Class, Diatomez. Order, Diatomales. This order contains a large number of genera and species both living and fossil. Diatoms can always be found forming brown scums or sediments on the bottom of ponds, creeks, ditches, etc. 2 * 18 LABORATORY OUTLINES FOR GENERAL BOTANY. 1. Mount some sediment or water containing diatoms and study the different species present. 2. Under high power, draw six different species, representing them from two to four inches long. They are unicellular plants with .two cilicified valves or shells which fit together like the lids of a pill-box. Represent carefully the mark- ings on the shell. In some species the ends and central portion of the valves are marked by nodules and these points are connected by a rib or suture called the raphe. These can be seen from the valve view. 3. Notice the greenish, yellow or brown chromatophores, the nucleus, and the cytoplasm. How are the cell organs arranged? 4. Look for chains or filaments of diatoms; also for stalked forms. 5. Study dividing forms. Some species conjugate. Look for such forms. 6. Study the movement. Does it have any relation to the field of the micro- scope, or the intensity of the light in the field? Describe. What is the cause of the motion? Remember that the motion is magnified under the microscope. How long does it take a diatom to pass across the diameter of the field? 7. Isthmuia sp. Scrape specimens of Isthmia from dry, red or brown alge or study from mounted slides. Isthmia can usually be obtained -from dry alge collected on the California coast. Draw a specimen from the girdle view, show- ' ing the valves and details of the markings. Notice that the individuals are of very different sizes. Draw one showing the valve view. Draw an individual in process. of division. Describe how the valves fit together, how new valves are formed, and what is the character of the valves of the two individuals resulting from a division. Explain the cause of the difference in size. 8. Fossil diatoms. Study material from the Tertiary deposit of Richmond,. Va. Place a fragment of the diatomaceous earth in a small bottle of HCl, crush gently and mount in water, or use prepared mounts. Draw three different species. 9. Note.— Diatoms, on account of the great number of forms, make a good study in variation. There is a great variety of patterns without very much advance in structure or life cycle—horizontal evolution. Is there any special advantage in the great variety of fantastic markings on the valves? XII. Clostérium sp. Phylum, Zygophyta. Class, Conjugate. Order,, Desmidiales. Family, Desmidiacee. Desmids are quite common in ponds and lakes and species of Closterium can usually be found in the sediments at the bottom, on submerged water plants, or in large masses floating on the surface. Sometimes Closterium is very abundant in watering tanks, forming large, green, floating flakes. 1. Mount in water and observe the large bright green, unicellular plants. which are more or less curved or crescent-shaped. 2. Draw an individual under high power, showing the cell wall with trans- vrese striations in the central region, the two large chromatophores (chloroplasts ) with highly refractive bodies (pyrenoids), the large nucleus with nucleolus in the central, clear space, and the peculiar vacuoles at each end. Notice the dancing, crystaline granules of calcium sulfate in the vacuoles. (Brownian movement). Descibe in detail, noting especially the symmetrical halves of the cell. 3. Notice the streaming of the cytoplasm between the large chloroplast and’ the cell wall. Trace the current around the end of the cell. 4. Look for dividing specimens. Draw and describe. ®. Search for conjugating individuals and for zygospores. LABORATORY OUTLINES POR GENERAL.BOTANY. 19 XII. Spirogyra sp. Phylum, Zygophyta. Class, Conjugate. Order, Zygnemales. Family, Spirogyracee. Spirogyra grows in stagnant water or slowly flowing streams, forming floccu- lent, floating masses of a bright green color which are slimy to the touch. It may be collected at any time but more commonly it conjugates in late summer and autumn. Some of the species will conjugate if brought into the laboratory and placed in an open dish of pure water. Metal is very injurious to Spirogyra. 1. Study naked eye characters, noting that the mass is made up of slender free threads or filaments. 2. Mount a few filaments in water and examine under low power. Notice the cells with spiral chromatophores (chloroplasts.) Shape of the filaments and cells? Count the number of cells across the cover-glass (# inch across). How many? Measure a long filament and estimate the number of cells it contains. 3. Draw part of a filament under low power showing ends, cells, and chro- matophores. Any difference between the two ends? Describe. 4. Under a high power, draw a cell showing the wall with mucilaginous sheath, spiral chloroplasts, pyrenoids, nucleus, and nucleolus. How is the nucleus connected with the other parts? 5. Draw part of a chloroplast showing details of the margin and the pyre- noids. 6. Treat with salt solution. Draw and describe what takes place. 7. Study the conjugation from fresh material, or if this is not at hand, from material preserved in copper salt solution or from mounted slides. Notice two filaments side by side and that all the zygospores are in the cells of one filament, while the cells of the other filament are empty. This indicates_a slight differ-_ entiation of sex individuals. Draw a piece under low power, showing a number of conjugated cells. 8. Draw two conjugated cells showing all details carefully, especially the zygospore or zygote and the conjugation tube. 9. Draw two cells in which the contents of one cell are passing thru the tube. 10. Draw two cells in which the two rounded processes from the side have just ete. oi met. 11. Draw two cells in which the’ two No processes are just beginning to develop. [le = 12. Describe fully the process of conju- Seer gation as observed above. C 13. Look for cases of parthenogenesis; a | we either with a spore in one cell and a distorted Co. protoplast in the other, or with a spore in each of the conjugated cells. 14. Make a diagram in the notes show- > ing the life cycle by means of diagram- fFyg 3.— DracRAM or ANCESTRY. matic figures of the plant, cells and spores. 15. Make a diagram showing the ancestors of one spore for five generations; take no account of vegetative propagation or of the possible close relationship of conjugating individuals (see Fig. 3). Compare with Pleurococcus. How many ancestors have you yourself had in twenty generations or about 600 years? 20 LABORATORY OUTLINES FOR (GENERALE SB OW Nae ‘A SERIES OF FORMS TO. ILLUSTRATE THE EVOLUTION Of XIV. Sphaerélla pluvialis (Flotw.). (Hematococcus). Phylum, Gonidiophyta. Class, Protococcee Order, Volvocales. Family, Chlamydomona- dacez. | Spherella may be found growing in rain water, drain tiles, roof gutters, pools, or ponds. It is unicellular and green in color or sometimes a bright red. If a culture is once obtained, it may be preserved on a limestone rock or in a glazed earthen jar. Put the rock into the water containing the alga and after some time take it out and lay it away. Whenever material for study is required the rock need only be placed in fresh rain water, when a new crop will soon: appear. ; 1. With a medicine dropper mount some water containing Spherella and examine under low power. Under high power study the large, green or red spherical cells in the resting condition. Draw. Notice the green and red coloring matters — chlorophyll and hematochrome. 2. Draw an individual divided into two, and one divided into four or eight cells. How does the division take place as regards the cell wall. Compare with RLO-O-G O-@<& Ms sk ae p oe | i 9-8 Fic. 4.— Lire Cycle oF SPHRELLA. Pleurococcus. Look for an individual in which the four cells are ready to break thru the old cell wall. The four cells form four free-swimming Sphzrellas which have very loose cell walls. 3. Study the active individuals. Describe the shape, color cell contents (especially the chloroplasts and pyrenoids), and the flagella. The flagella branch out from a clear body in the pointed end of the cell and pass out thru two ex- tremely minute openings in the cellulose wall. 4. Study and describe the movement. Which end is directed forward in swimming? How long does it take an individual to pass across the diameter of the field? Suppose the diameter is three-tenthes of a millimeter, how long would it take the plant to travel thirty centimeters or one foot? Is the motion rapid or slow? How many times its own diameter does an individual move in one second? 5. The flagella and other parts may be seen more clearly by adding a small drop of iodin solution to the water at the edge of the cover-glass. What hap- pens? How long are the flagella, compared with the size of the body? Look for the nucleus. Notice the protoplasmic strands which pass from the cell-body- to the wall. 6. Notice the division of labor in the organism and designate the function of the following organs: a, cell wall; b, flagella; c, chloroplasts. LADOMANOKY OCULEINES FOR GENERAL BOTANY. Oil 7. Make a diagram in the notes showing the life cycle of Spherella when reproduction takes place by the formation of non-sexual zoospores. (See Fig. 4a.) 8. Sexual stage. Look for individuals divided into eight or more cells. Draw. When these escape they are smaller than the four and at first have no cell walls. These zoospores are said to conjugate and form zoozygospores. Care- ful observations should be made in order to discover such a process. In case conjugation takes place the life cycle during this stage may be represented as in Fig. 4b. XV. Pandorina morum (Muell.). Order, Volvocales. Family, Volvocacee. Pandorina occurs in small pools of water, and is often very abundant in summer, coloring such pools a bright green. The individuals consist of a free- swimming colony of sixteen cells, and are more or less globular or oval in outline. 1. Mount some of the colonies in water and examine under low power. Notice the active movement. Draw a colony under high power. If they cannot be followed because of their active movements, add a drop of carbolic aicid water. 2. Notice the details of an individual cell of the colony; the two flagella, the red eyespot, the transparent spot in the outer end of the cell, and the chlor- oplast with a pyrenoid. 3. Study and draw colonies in stages of division. Each of the sixteen cells divides until each forms a group of sixteen new cells, then the gelatinous envelope dissolves and the sixteen daughter colonies are set free. This is the normal method of vegetative propagation. 4. Sexual reproduction. . Look for colonies in which the cells are separating as isolated zodspores. These are the gametes which are very much alike, but are of various sizes. 5. Watch for conjugating forms. Conjugation takes place between two gametes of equal size, or between a larger and a smaller one. The process is complete in a few minutes. Draw stages observed, and also mature zygospores. The difference in size of the conjugating gametes is of special importance, since it is the first step in the evolution of two specialized gametes, the odsphere and spermatozoid. 6. Note.— Pandorina is well preserved in water with carbolic acid, and large quantities may be collected at the proper season showing the various stages of the life cycle. Cultures can also be obtained in the laboratory from dry zygospores. XVI. Eudorina élegans, Ehrb. Family, Volvocacee. Eudorina frequently occurs in summer in pools of rain water, in ponds, and in marshes. The colonies are hollow, free-swimming bodies, more or less spheri- cal in shape, usually consisting of thirty-two cells which are considerably sepa- tated from each other. 1. Mount a drop of water containing the organism and examine under low power. Under a high power draw a single colony, showing the arrangement of the cells. 2. Draw a single cell, showing the two flagella, the red eyespot, and the chloroplast with a pyrenoid. 3. Vegetative propagation. The individual cells divide into sixteen or thirty- two new cells, and these escape as daughter colonies the same as in Pandorina. Draw a colony showing daughter colonies, and describe. 2 29 LABORATORY OUTLINES FOR “GENERAL, BOPANNe 4. Sexual reproduction. The colonies are either unisexual or hermaphrodite. Draw a colony showing antherida (spermaries), consisting when mature of plates of sixty-four small cells each, which develop into male gametes (spermatozoids). Draw and describe free-swimming spermatozoids. 5. Draw a colony containing female gametes (odsperes). The colony with oospheres differs very little from the ordinary vegetative colony. Watch for spermatozoids swarming about the female colonies. 6. Draw and describe the ripe, red-colored odspore. 7. Note.—Eudorina shows a considerable advance in sexual development over Pandorina. The female gamete (odsphere) has become stationary, but still retains its flagella at first, and does not divide. The male gametes (spermatozoids) are formed by the repeated division of the cells of the colony. They are very small in comparison with the female cell, swim about freely in the water, and have lost their chlorophyll. XVII. Vélvox globator L. Family, Volvocacez. This alga is of such size that its spherical, free-swimming body can easily be seen with the naked eye. In summer and autumn it can frequently be found in fres water ponds and lakes. 1. Take up some of the spherical colonies with a large-mouthed medicine dropper or a glass tube and, having formed a little chamber on the slide with a xylonite ring or with paraffin, mount and study under low power. Note the rotating movements of the hollow, spherical organism. 2. Draw a colony showing the numerous cells and some daughter colonies, which appear as darker green spherical masses of various sizes. 3. Under high power, study a single colony. About how many cells in a colony of average size? Draw a few cells, showing the cell walls, the proto- plasmic strands, connecting the cells (protoplasmic continuity), the chloroplast, the red eyespot, the pulsating vacuole, and the two flagella of each cell. The ~ flagella will be more distinct after staining with iodin. 4. Describe the devleopment of a daughter colony from one of the cells of the mother colony. Look for an opening (the pore) in one side of the young colonies. What advantages may there be in the hollow spherical form? 5. Sexual reproduction. The colonies are hermaphrodite, developing both sexual organs—the ovaries or oo0gonia and the spermaries or antheridia —in late summer or autumn. Draw an antheridium. This represents an enlarged cell of the colony which has divided into a large number of elongated cells arranged like a bundle of asparagus shoots. 6. Draw an odgonium, projecting into the cavity of the colony, showing the enlarged odsphere. Draw a ripe odspore, showing the thick wall with peculiar angular points on the surface. 7. Note.—In Volyox complete sexuality has been attained with the normal conditions of the sexual cells (gametes). It will be noticed that the plant is hermaphrodite, and this is the more usual condition in the lower plants. XVIII: Vauchéria séssilis Vauch. Phylum, Gontdiophyta, =9@lass, Siphonez. Order, Vaucheriales. Family, Vaucheriacee. This alga grows as a lax, green, felt-like layer on the surface of moist soil, and is especially common on the surface of pots in greenhouses, and may here be in fruit at any time of the year. Other species may be found in ponds. EABOKAN OKVAOULEINES FOR GENERAL BOTANY. 22. 1. Describe the naked eye characters, noting the coarse cylindrical filaments. 2. Under low power draw an entire filament showing the branches, the tips, and the clear or decaying part of the back end. Note the absence of transverse walls. Such a plant body is called a cenocyte. The protoplasts are not separated by walls. How does the filament grow? 3. Under high power draw a short piece of a young filament, showing details. —shape, vacuole, arrangement of protoplasm, chloroplasts, and oil drops. Posi- tion of chloroplasts? How can you tell that the filament is cylindrical without. seeing a cross section? 4. Draw several chloroplasts. Shape? Draw some in stages of division. Describe. Look for movement of the protoplasm. ‘Numerous nuclei are present in the cytoplasm, but these are not visible without special staining. 5. Study the sexual organs, the antheridium (spermary), and oogonium (ovary). They are usually side by side. Draw carefully and describe. Notice the septa which separate the sexual organs from the main filament. Vaucheria is hermaphrodite, having male and female organs on the same individual. 6. Draw the oosphere (unfertilized egg); also some spermatozoids in the autheridium. Look for free-swimming or escaping spermatozoids; also for sper-— matozoids entering the oogonium. - 7. From the union of the two gametes an oOspore or zygote is formed. Draw a ripe odspore showing the thick wall and more or less hyaline contents. Describe. Note that fertilization, in this case, is not the stimulus for further development but is followed by a resting stage. 8. Contrast the two sex cells (gametes) as to size, motion, and nutrition. How is an oospore different from a zygospore?. Would there be any advantage in this? 9. Special vegetative propagation by means of volvox-like colonies (com- pound zoospores), produced in the ends of the filaments, may be obtained as fol- lows: Place a mass of Vaucheria in a porcelain dish, in water, and expose for a. few days in the window until small Vaucheria plants are found floating on the surface. Examine very early in the morning and the volvox-like colonies may be seen escaping from the swollen ends of the filaments. In order to observe the colonies later in the morning, cover the dish, the evening before observation is to: be made, so that the plants will be in absolute darkness until shortly before the material is to be studied. Study and draw. Describe in detail the formation of the compound zoospores and how they develop into new Vaucheria plants. Might: this process indicate some relation of the ancestors of the Vaucheria to the Vol- vocacee ? 10. Make a diagram in the notes, showing the life cycle of Vaucheria. (See Fig. 5.) 11. In the notes, make diagrams of the two gametes of Spherella, Pan- dorina, Eudorina, and Volvox, and describe how these may indicate stages in the evolution of perfectly developed odspheres and spermatozoids. 12. Notre.— The odsphere and spermatozoid are highly specialized cells, the first representing nutritive qualities, the second the active qualities. A union of the two must result in a very perfect reproductive cell. The development of sexual individuals appears to be along the same lines as indicated in the sexual cells. Maleness or femaleness is not an hereditary character or factor, but a condition and often depends on the environment present during the germination of the spore or the development of the embryo. In some of the intermediate plants the sexual’ development can be controlled while in the higher groups the sex of the gametophyte: is always determined in the spore. D4 LABORATORY OUTLINES FOR GENERAL BOI TWO PECULIAR CENOECY DIC COLONiS: XIX. Pediastrum pertusum Kuetz. Phylum, Gonidiophyta. Class, Hydrodictyee. Order, Hydrodictyales. Family, Hydrodictyaceze. This beautiful alga is found along with other species of the same genus in the sediments at the bottom of ponds and creeks, and is especially abundant in the plankton of fresh water lakes and bays in summer and autum. It is a flat colony of cells which develop into cenocytes. 1. Mount some of the sediment containing Pediastrum in water and study under high power. Draw two of the plate-shaped colonies—one with sixteen cenocytes and one with thirty-two or more. Notice the difference between the marginal cenocytes and those in the interior. Note also the chloroplast and one or more pyrenoids. 2. Look for colonies in which the cells in each cenocyte have separated, preparatory to the formation of a new colony. 3. Draw a colony in which some of the cenocytes are empty, each empty shell having a slit-like opening thru which the daughter colony escaped. Fic, 5.—LireE Cycle oF VAUCHERTA. XX. Hydrodictyon reticulatum (L.). Water-net. Family, Hydrodictyacee. The water-net forms a large body, which is common in summer and autumn in ponds and canals. It may often be collected in great quantities along the grassy banks of ponds in city parks. The body of the alga is made up of a very great colony of cylindrical cenocytes arranged in the form of a sack-like net. 1. Examine a large plant and describe the naked-eye characters. If fresh material is at hand, place the plant for some time in direct sunlight and note the - bubbles of gas collecting in the nets. Of what use? What is the gas. 2. Draw a small portion of a young net under low power, showing how the meshes are formed by the joining of a number of cenocytes. Describe. 3. Under high power draw a single cenocyte, showing the chloroplast and numerous pyrenoids. 4. Vegetative propagation. Study and draw a large cenocyte of an old net in which the cells are developing a daughter net. PABORALORY QULEINES POR GENERAL BOTANY. 25. A NUMBER (OF COMPLEX. ATTACHED FORMS. XXI. (a) Caulérpa crassifolia (Ag.). Phylum, Gonidiophyta. Class, Siphonee. Order, Caulerpales. Family, Caulerpacee. This is a large, much branched, cenocytic plant which reproduces by the branching of the thallus. The creeping, rhizome-like portion ‘develops leaf-like, lobed branches, and continues to grow in front while it dies behind. It grows. within the tropics on sandy ocean shores at a depth of several meters. Preserved material will be necessary. | 1. Sketch the entire plant showing the creeping “rhizome,” the leaf-like lobed branches, and the branching root-like hold-fasts. 2. Mount part of a leaf-like branch and examine under low and high power. Note that there are no septa or walls in the plant body. Note the chloroplasts. Draw. Ba 3. Examine a cross section of the cylindrical “rhizome” under low and high power. Note the numerous branched and anastomosing cellulose braces stretched across the cavity. Draw. 4. Note.—Caulerpa represents a plant which has developed the extreme type of a cenocytic body. The body takes on a shape suitable to its activities, by its branching habit affording sufficient surface for absorption and photosynthesis. The rigidity of the body is secured thru the complicated system of braces rather than by cell walls. Compare Note 6 on Pleurococcus. (b) Botrydium granulatum (L.). Phylum, Gonidiophyta. Class, Siphonee. Order, Botrydiales. Family, Botrydiacezx. This alga is common in summer and autumn, especially after heavy rains, on wet ground in fiields and around ponds, where it forms little green pearshaped bodies with rhizoids extending into the soil. 1. Draw a thallus under dissecting microscope or low power, showing the pear-shaped zrial part and the branching rhizoids. Describe, noting the division of labor in the plant body and that it is a cenocyte. 2. If material is at hand study some plants which are producing zoospores. Examine under high power and describe. How many flagella? XXII. Cladéphora sp. Class, Siphonee. Order, Cladophorales. Family, Cladophoracee. Species of Cladophora are commonly found in flowing water. They appear as large, dark green extensively branched tufts, attached to rocks and pieces of wood. These alge are also abundant along the shores of lakes, where they may often be found attached to objects which are exposed to the action of waves. They may be obtained at any season. i. Describe the naked eye characters; size, mode of growth, color, habitat, etc. Note the differentiation of base and apex. 2. Mount a small branch of the thallus in water and study under low power. Draw. 3. Under high power study the stages showing the development of a branch Make four drawings showing four general stages: (1) a small bulging out of the cell wall on one side below the septum; (2) a short branch with the proto- plasm still connected with the parent cenocyte; (3) the branch cut off from the parent cenocyte by a septum; (4) a branch divided by a transverse septum into two cenocytes. 26 LABORATORY OUTLINES FOR GENERAL BOTANYS 4. Draw a single cenocyte or division between two cross walls, showing the irregular chloroplasts and the pyrenoids. Notice the large central vacuole. Apply salt solution and note the effect. Numerous nuclei are present, but these can probably not be distinguished from the pyrenoids without special staining. Stain with iodin solution and draw. It is a Cenocytic plant with numerous transverse walls, but the wall do not represent cell divisions. How and where do the branches always originate? 5. Draw and describe an empty cenocyte or zodsporangium from which zoospores have escaped. Where is the opening (ostiole) always formed? 6. At certain times zoospores may be seen forming and escaping into the water. This frequently occurs in material which has been kept in water in a warm room. Draw a cenocyte in which the zoospores are developing. 7. Draw a cenocyte in which the zoospores’ are fully formed. Look for zoospores in the act of escaping thru the ostiole. 8. Study and draw free-swimming zoospores, showing the chloroplast and red eyespot. To make the two flagella visible treat with iodin solution by placing a drop on the slide beside the cover-glass and letting it mix slowly with the water of the mount. 9. Draw a zodspore which has begun to. develop into a new filament. The embryo developing from a free spore is called a sporeling. 10. The zodspores (planogametes) of some species are said to conjugate. Look for such a process. XXIII. Ulothrix zonata (W. & M.). Phylum, Gonidiophyta. Class, ‘Confervee. Order, Confervales. Family, Ulothrichacee. This Ulothrix is a small, unbranched, filamentous, green alga which usually grows in running water, attached to sticks and stones. It may be found in slow- flowing streams, in watering troughs, or in fountains. Collect the material and place it in a shallow dish in about two inches of water, and in a day or two, after the water has evaporated somewhat, large non-sexual zoospores and sexual gametes will probably be forming. Study the fresh material and preserve some for further use. 1. Mount some of the filaments containing the basal cells (hold fasts) and study under low power. Draw. 2. Under high power draw the holdfast, the terminal cell, and two or three of the central cells; showing the wall, the chloroplast, and the nucleus. Describe these parts. 3. Non-sexual zoospores. Examine a filament in which the cells are form- ing either one, two, or four zodspores each. Observe how they escape by a lateral opening in the cell-wall. Draw and describe. These spores have four flagella and a pulsating vacuole. Draw an empty cell. 4. Sexual reproduction. Study a filament in which the cells have developed a large number (eight, sixteen, or thirty-two) small gametes of equal size (iso- gametes). Draw part of the filament showing some cells empty and some with gametes. The gametes have only two flagella. 5. Observe the conjugation of the planogametes to form zoozygospores. Draw and describe. In order to bring out the flagella more clearly, stain with iodin solution. If the gametes do not conjugate some may round themselves off and become resting spores. This is a case of parthenogenesis. 6. Notre.— When the zygospore germinates it does not develop a new fila- mentous plant, but gives rise to a number of cells which develop as non-sexual a PABOKRATORY OUTLINES POR GENERAL BOTANY. 27 zoospores, and these escape and produce the filamentous plant. Ulothrix, there- fore, along with many other thallophytes, has what is known as an alternation of generations. XXIV. Ectocarpus confervoides (Roth.). Phylum, Phzophyta. Class, Pheosporee. Order, Ectocarpales. Family, Ectocarpacee. This alga has a branching filamentous frond and grows in summer attached to other alge or to submerged objects along the seashore. 1. Note the form of the plant and mode of attachment. Draw. 2. Examine under low power. Are the cells in a single row? Draw a branch. 3. Study and draw a plurilocular sporangium; also draw one of the unilo- cular sporangia which are usually developed somewhat earlier than the plurilocular ones. The unilocular sporangia produce non-sexual zodspores and the plurilo- cular sporangia produce planogametes. XXV. Ftcus evanéscens Ag. Phylum, Pheophyta. Class, Cyclosporee. Order, Fucales. Family, Fucacee. This brown alga is common along the Atlantic coast. It may be obtained from dealers in botanical supplies, and preserved in alcohol or other solutions, or they may be dried and soaked in water when needed. Various species of Fucus may be found fresh at fish stores in large cities, these plants often being used as packing. The thallus is a large, flat, dishotomously branching frond of a dark brown color, attached to various objects by means of a disk-like holdfast. 1. Place the plant in a plate of water and draw the large thallus. Describe. Note the holdfast, the flattened dichotomous frond, and the thicker central region forming a sort of midrib. Note also the swollen tips of the branches Giceepracies)- covered with numerous dot-like projections. 2. Find the growing points of the thallus in branches which do not have receptacles. Note the emarginate apices which have a slight groove lying in the plane of the thallus. Draw under low power. The initial cells are at the bottom of this groove. How are the branches formed? 3. Cut thin cross sections of a branch of the thallus with a razor, mount, and examine under low power. Draw. Note the outer, denser, cortical layer, and the loose, inner region, with elongated branched filaments and much mucilage. 4. Cut thin cross sections of a receptacle, mount, and examine under low power. Note the conceptacles, cavities opening by means of ostioles on the ex- terior. Sketch the entire section. 5. Select a favorable conceptacle and draw showing the ostiole, the wall of the conceptacle, the sterile hairs, the large, dark-colored ovaries (oogonia) of oval form, and the small yellowish spermaries (antheridia) situated on branched hairs. 6. Under high power draw and describe a single antheridium showing cells developing into spermatozoids. About how many sperms does an antheridium produce? 7 (. Draw and describe an oogonium containing the eight ripe oospheres. 8. Compare the egg and sperm cells. About how much larger in volume is one than the other? 9. If fresh material can be obtained, study the spermatozoids and oospheres after their escape from the sexual organs. Take a plant with mature receptacles from sea water and expose it to the air for several hours. Mount some of the exudation, which appears at the ostioles of the conceptacles, in sea water, and 28 LABORATORY OUTLINES FOR GENERAL BOTAND, examine under high power. Notice the large spherical oospheres and the small motile spermatozoids. Study the process of fertilization, and describe. Draw an oosphere surrounded by spermatozoids. The discharge of the egg from the ovary into the water is a very unusual phenomenon in the plant kingdom. Compare with Vaucheria and Volvex. FOUR PHYCOMYCEERBES SHOWING VARIOUS, BART AWS XXVI. Mucor stolonifer Ehrenb. Black Bread Mold (Rhizopus nigricans. ) Phylum, Mycophyta. Class, Zygomycete. Order, Mucorales. Family, Mu- coracee. This fungus can nearly always be obtained by placing a piece of old bread for several days in a moist chamber. An ordinary glass or jar with a cover will do very well for making the culture. Enough water should be added to keep the bread moist without soaking it. The fungus forms a white flocculent mass of cottony filaments (the mycelium made up of, hyphae) over the surface of the bread and later also spreads out over the walls of the glass. Some of the hyphe w:ll be seen to rise vertically into the air and end in rounded black heads. These are the sporangia containing the non-sexual- spores. 1. Describe the naked eye characters noted above. Notice habitat and color. Notice also (1) the hyphae passing down into the substance of the bread, (2) the horizontal stolen-like hyphae, and (3) the upright sporangiophores. 2. Cut off a flake of the young hycelium with a pair of scissors and mount in water, taking great care not to injure the delicate hyphae. Study under low power and draw some of the hyphe showing mode of branching. 3. Mount carefully and under high power draw part of a hypha and describe. Any transverse septa (cross walls)? If not, what kind of a fungus is it? (Com- pare with Vaucheria.) How does this plant differ from the green alge in general? Difference in mode of obtaining food? Why is this plant called a saprophyte? 4. Study and draw a cluster of sporangiophores showing the rhizoids at the base and the sporangia at the tips. The best are those taken from the walls of the dish. Color? Draw a single unbroken sporangium showing the columella on the inside, and the non-sexual spores. Do not mistake the columella of a broken sporangium for the entire body. Describe the structure of the sporangium. What does the columella represent? The sporangia burst readily because of the presence of an intermediate substance which swells readily in water. Of what use is this? 5. Draw and describe the non-sexual spores. Color? About how many in a sporangium ? 6. This plant has a partial development of sexuality, and under suitable con- ditions, if the female and male (or + and —) strains are growing together, pro- duces zygospores. If any of these are at hand or material from another species, study and draw showing the following stages. a. Two neighboring branches of the mycelium, which are about to conjugate and which are in contact. b. The stage in which the two branches have fused. c. The stage in which transverse septa are formed, cutting off the apical part of each conjugating branch. Any difference between the conjugating branches in size or contents? LABORATORY OUTLINES FOR GENERAL BOTANY. 99 d. The absorption of the wall separating the conjugating tips and the subse- quent mixing of the two cenocytic protoplasmic masses. ce. The mature zygospore suspended between the two branches. XXVII. Emptsa mtscae Cohn. Fly-cholera Fungus. Phylum, Mycophyta. Class, Zygomycete. Order, Entomophthorales. Family, Entomophthoracez. . This fungus grows on the common house fly (Musca domestica). In the autumn dead or dying flies attacked by this fungus may be seen with greatly swollen abdomens of a white color. Specimens may be preserved in alcohol. 1. Study a fly recently killed by this fungus; under low power without a cover-glass. Note the bands of short white hyphze (conidiophores) protruding from between the black segments of the abdomen. Draw and describe. 2. Tear open the abdomen with needles and mount the white contents and some of the conidiphores in water. Examine under high power. Notice that the mycelium has nearly absorbed the contents of the fly’s abdomen. 3. Draw some of the conidiophores with conidia still attached; also draw several of the conidia. Describe. Is the fungus a parasite or a saprophyte? XXVIII. Saprolégnia sp. Water mold. Phylum, Mycophyta. Class, Odmycete. Order, Saprolegniales. Family, Sap- rolegniacee. This fungus can usually be obtained by placing dead flies in a dish of spring or pond water. After about five or six days the hyphe of the fungus may be seen protruding from the body of the fly. On the tips of these hyphze sporangia are developed which discharge numerous zoospores. 1. Notice the fly in the water, surrounded by a halo produced by the mycelium of the fungus. 2. Mount some of the mycelium in water and examine under low power. Draw a branch under high power, showing the granular protoplasm and a terminal sporangium developing zoospores. Draw a branch showing an empty sporangium. 3. Study and draw free-swimming zoospores. Oospores may also be present. XXIX. Plasmépara viticola (B & C.). Downy Mildew of ‘Grape. Class, Oomycetae. Order, Peronosporales. Family, Peronosporacee. This mildew causes a destructive disease of the leaves and young shoots of the cultivated grape. The infected leaves may be collected in spring or summer and preserved in 70 per cent. alcohol or dried and kept in paper boxes. Conidial stage. 1. Examine a leaf carefully under the low power, without cover-glass. On which side do the conidiophores appear? 2. Carefully scrape off some of the conidiophores with a needle or scalpel, mount in water, and examine under low power. Under high power draw one of the much branched conidophores. If dry material is used the conidia will probably all have dropped off. One is developed at the tip of each peg-like branch of the conidiophore. 30 LABORATORY OUTIINES FOR GENERAL BO PANae 3. Draw several conidia and describe shape, size, and color. 4. From alcoholic material cut cross sections of a part of a leaf containing the fungus, mount, and under low power note that the conidiophores come out im bunches thru the stomata of the leaf. Draw. . 5. To what physiological group does this fungus belong — parasite or sapro- phyte? Describe its mode of life so far as studied. EXAMPLES OF THE HIGHER ALGZ. XXX. Chara sp. Phylum, Charophyta Class, Charez. Order, Charales. Family, Characez. Stoneworts. The stoneworts are alge which are found growing in the bottom of ponds, lakes, or slowly flowing creeks and rivers. They are of considerable size and are usually covered with an incrustation of lime. They contain numerous branches arranged in whorls and are firmly fixed in the mud by means of rhizoids. Charas grow very readily in an aquarium and may be kept in a healthy condition all winter by simply placing the plants into a glass jar of water and keeping them near a south window. 1. Sketch an entire plant and describe the naked eye characters. Notice the odor, the nodes and internodes, and the brittleness of the filaments. 2: Mount the base of a plant in water and examine under low power. Draw and describe some of the branching rhizoids. Study the rotation of the proto- plasm under high power. 3. Mount the treminal part of a young branch, being careful so as not to crush the brittle lateral branches. Examine under low power and draw the terminal bud. Notice the great internodal cells covered with a cortical layer and the whorls of lateral branches. 4. Draw a cell of one of the branches without a cortical layer, showing the incrustation of lime. 5. Under high power draw a part of a cell, showing the chloroplasts. How are they arranged? Draw several in stages of division. How do they divide? 6. Study the rotation of the cytoplasm in the large cells of the branches and describe. How does it differ from that in the cells of Philotria? Why are the chloroplasts arranged in rows? Note the movements in opposite directions on either side of the neutral line. Is the direction of rotation the same in all the cells. lord (7. How is the cortical layer developed? In order to determine this, young branches should be obsreved. Draw a cross section of the main stem. Note the short projecting cells which roughen the surface. 8. The sexual organs are produced during summer and autumn. Study fresh material, or if this is not at hand, material preserved in alcohol or copper solution. The spermaries (antheridia) and ovaries (oogonia) are situated on the lateral branches. Draw. Notice the five spiral branches which cover the ovary. How does this ovary differ from that of Vaucheria? The spermaries are globular organs which are red in color when fresh. Is this plant hermaphrodite or uni- sexual? If the incrustation of lime is too thick remove it with Perenyi’s fluid. 9. Crush a ripe spermary under the cover-glass and draw one of the numer- ous filaments inside. The small cells of these filaments contain the spermatozoids. Draw a single cell showing a mature spermatozid. How many cells in a single filament? Suppose the spermary contains 8x6x4 filaments, how many spermao- WABOKALORY “OCLTEINES POR GENERAL BOLANY. 31 zoids would there be produced in each spermary? How many spermatozoids for each oosphere or egg? 10. Draw an ovary containing a ripe oospore. Explain the structure of the entire body. Crush the oospore and note the starch. Treat with iodin. 11. Study the proembryo, from which the normal Chara plant develops as a lateral bud. Draw and describe. Proembryos may be obtained by placing plants with mature spores in a glass jar of water and keeping them over winter. In the spring the embryos will be found at the bottom. 12. Make a diagram showing the life cycle of Chara. Compare with Vau- cheria. XXXI. Batrachospérmum monilif6érme Roth. Phylum, Rhodophyta. Class, Florideez. Order, Nemalionales. Family, Hel- minthocladiacee. Batrachospermum is an alga of considerable size which can be found attached to stones in fresh water rivulets and creeks. 1. Spread out the frond of the alga in water in a porcelain plate and sketch the entire plant. 2. Mount some of the branches in water, crushing them considerably under the cover-glass, and sketch under low power. 3. Under high power draw one of the lateral branches coming out from the nodes. Note the oval cells and the bristle-like projections on some of the terminal cells. 4. In a young, main branch study the branches which pass down from the base of the nodal branches and form a loose cortical layer. How does this com- pare with the cortical layer in Chara? 5. Crush some of the older branches under the cover-glass by pressing and rubbing carefully over the surface with the handle of the needle and study the ovaries. These are situated on the lateral branches, and each consists of a thick- ened hair-like process (trichogyne) and a bulbous base (trichophore) containing the odsphere. Draw. 6. Study the spermaries, which are single terminal cells, each of which de- velops a single spherical male gamete (spermatium) without flagella. Draw a spermary and a free floating spermatium. 7. Draw an ovary which has one or more spermatia attached to the trichogyne. 8. Draw a sporocarp under low power. This is a sperical cluster of branches which develops from the fertilized egg. 9. Under high power draw a nonsexual carpospore at the end of one of the branches of the sporocarp. 10. From the foregoing study it will appear that Batrachospermum possesses a sort of an alteration of generations. Besides this, it has another interesting stage. When the carpospore germinates it gives rise to a peculiar filamentous proembryo or protonema, formerly known as the chantransia stage, from which the normal Batrachospermum plant develops as a lateral bud. Protonemal plants should be collected showing various stages in the development of the Batrachos- _ permum bud. The protonemal (chantransia) filament can reproduce itself by means of non-sexual spores developed on the tips of its branches. This is a case of reproduction known as pedogenesis, since the process is accomplished while the plant is in the immature condition. If material is at hand, draw and describe the chantransia filaments and spores. 29 LABORATORY OUTLINES FOR GENERAL BORANY XXXII. Polysiphonia variegata (C. Ag.). Phylum, Rhodophyta. Class, Floridee. Order, Ceramiales. Family, Rhodo- melacez. Polysiphonia grows in abundance on rocky sea coasts. The plants may be found in summer as purplish-brown tufts, a few inches long on other larger water- plants, or on piles and stones. Preserved material may be used by those living away from the seashore. 1. Spread out a frond in a porcelain plate and sketch the entire thallus. Note the holdfast, if present, and the mode of branching. 2. Mount a branch and draw under low power. Note that the body of the thallus consist of successive tiers of cells, each tier consisting of a central cell, surrounded by a layer of cortical cells. & “Under high power draw a single tier of cells. Crush the thallus a) iitele and note especially the large protoplasmic strands (protoplasmic continuity) which run from the central cell to the several cortical cells of the tier. Note also, the strands connecting the cells of a tier with those of the tiers above and below. 4. Cut cross sections of the thallus, mount, and study under high power. The sections may easily be obtained by chopping up a moist branch on a piece of paper with a sharp scalpel. Draw, showing the arrangement of the central and cortical cells and also the protoplasmic connections. 5. Under high power study the tip of a young branch and draw. Notice the dome-shaped apical cell and a number of cells below. The lower ones are divided by longitudinal walls. How are the tiers and the cortical cells developed? From this it is evident that, altho Polysiphonia appears like a branched filament and continues to develop as such, it finally forms a true solid aggregate. 6. Nonsexual spore reproduction. Mount branches of plants containing tetra- sporangia (dark spherical bodies below the cortical cells) and draw under high power. Draw the spore tetrad and one of the mature spores. 7 (. Sexual reproduction. The spermaries (antheridia) are borne on delicate, colorless dichotomously branched filaments, which form tufts on the younger part of the frond; the ovaries (oogonia) are on short branches in the upper part, or the individuals may be unisexual. Mount branches containing spermaries and draw under high power. Note the slender tip of the branch which extends beyond the oblong spermary. Note also that the sexual plants have no tetraspores. 8. Development of the ovary. Mount branches containing young ovaries and under high power draw: (a) a short lateral simple branch showing one of the cells considerably enlarged and more or less spherical; (b) one in which this cell has divided by vertical walls into four cells; (c) one in which the inner cell of the tier of four has enlarged and divided into three or four cells by transverse walls, the upper one developing into the ovary with a basal trichophore and a slender trichogyne; (d) a young cystocarp showing the trichogyne protruding from the mass of cells forming the wall. 9. Draw a mature cystocarp, showing the more or less ovate-globose wall and the carpostome. 10. Crush one of the mature cystocarps and draw several of the dark colored, nonsexual carpospores. 11. Note.— Polysiphonia has an alternation of generations, since the spores of the cystocarp and the tetrasporic plant are homologous to the sporophyte of LABORATORY OUTLINES FOR GENERAL BOTANY. 33 higher plants. Note that the sporophyte is at first nursed by the gametophyte then produces a number of independent spores. Its life cycle may be stated as follows: spermatium Sexual plant or gametophyte Srephere fertilization — germination of oospore — carpospores —tetrasporic plant or sporophyte (nonsexual) — sporocyte — reduc- tion division — tetraspores — gametophyte, etc. 2 tHE HIGHER FUNGI AND LICHENS. XXXIII. Aspergillus herbariodrum (Wigg.). Common Green Mold. Phylum, Mycophyta. Class, Ascomycete. Order, Aspergillales. Family, Aspergillacez. This mold is exceedingly common on improperly canned fruit, on cheese and on decaying plants; especially on plants in press for the herbarium when the driers are not frequently changed. The conidial stage is of a greenish color while the ascosporic stage is of a bright yellow-orange to the naked eye. 1. \Conidial stage. Mount carefully in water and study under low power. Under high power draw a conidiophore with conidia. Describe. How are the conidia developed? Draw a piece of the vegetative mycelium, showing the trans- verse septa. 2. Ascus Stage. Mount some of the white mycelium around the margin of the yellow centre. Under high power draw some of the peculiar coiled hyphal bodies present. These represent the conjugating branchs, from which a fruiting body devolops. 3. Draw the mature fruiting body (ascocarp) under high power, from a mount of the yellow colored mycelium. Notice the asci containing ascospores. 4. Crush the ascocarps under the cover-glass and draw an ascus with spores. Describe the life history of the plant. The ascocarp may be compared in a general way with the cystocarp of Polysiphonia. XXXIV. Morchélla esculénta (L.). Morel. Class, Ascomycete. Order, Helvellales. Family, Helvellaceze. This edible morel is common in spring and summer in moist woods and shady hillsides. Specimens may be preserved in 70 per cent. alcohol. 7. Make a careful sketch of the entire, fleshy, fruiting body, representing the stalk and the deep-pitted pileus on whose surface the asci are borne. 2. Tease out a piece of the stalk and mount in water. Examine under high power and draw some of the mycelial threads. Note that the entire body is a spurious tissue of interwoven septate hyphe. Compare with Mucor. 3. Tease out a small piece of the pileus, mount, and study the asci. Draw. How many ascospores in an ascus? How do the asci open at the tips? XXXV. Uncinula: salicis (DC.). Class, Ascomycete. .Order, Perisporiales. Family, Erysibacez. This powdery mildew grows as a parasite on the leaves of various species of willow and can usually be found without difficulty in the autumn. It forms a white layer on the surface of the leaf in which minute black bodies are situated. It may be preserved in 70 per cent. alcohol or kept in a paper box. 1. Moisten a leaf with water and scrape off some of the mycelium con- taining the black bodies (cleistothecia). Mount in water and examine under low 3 34 LABORATORY OUTLINES FOR GENERAL 50 TyAiNee power. Under a high power draw a cleistothecium with appendages. Be careful to have one that is mature. 2. Draw a single appendage. Of what use are the appendages and the coiled tips? Draw a small piece of the mycelium showing the transverse wall in the hyphe. 3. Crush some of the cleistothecia under the cover-glass by pressing and rubbing carefully over the surface with the handle of the needle. Draw an ascus containing ascospores. How many asci in a cleistothecium? How many spores in an ascus? / XXXVI. Saccharomyces cerevisiae Meyen. Beer and Bread Yeast. Phylum, Mycophyta. Class, Ascomycete. Order, Saccharomycetales. Family, Saccharomycetacee. To obtain yeast plants in active, vegetative condition, take a piece of ordinary dry yeast cake and put it in a glass of water containing a small amount of sugar. Keep over night in a warm place. 1. Mount some of the water containing yeast plants and study under high power. Draw several of the large oval cells present; also a short, branched’ fila- ment of cells. 2. Notice the formation of new cells by process of budding. Draw a number of cells showing the several stages in the formation of a daughter cell. 3. Compare the size of a yeast cell with one of the bacteria present. 4. Stain with iodin solution. Notice, the yellowish-brown color of the yeast plants and the blue of the large starch grains of the yeast cake. Is there any starch in the yeast cells? - 5. Draw a large cell carefully, showing granules in the protoplasm and one or more vactoles. 6. Nore. Yeast plants produce alcoholic fermentation in saccharine solu- tions. Dry bread yeast is usually a form of the beer yeast, and is known as “surface yeast.” XXXVII. Ustilago zéae (Beck.). Corn Smut. Phylum, Mycophyta. Class, Teliosporee. Order, Ustilaginales. Family, Ustilaginacee. The corn smut may be collected in summer and autumn and kept in a dry condition in paper boxes. 1. Make a naked eye sketch of one of the large, black, smut nodules. On what parts do the smut nodules develop? | 2. Mount some of the black powder and study under high power. Draw a number of the small spores. These are usually known as chlamydospores, or teliospores. Describe the color, surface and shape. 3. Make a hanging-drop culture of the spores with dilute, boiled, stable- manure water. Smut spores germinate quite readily, but it is best to let them freeze before making the culture. Watch the germination from day to day and note the formation of the small promycelium or basidium which develops a number of hyaline basidiospores. Note that the smut plant is a parasite while the pro- mycelium is a saprophyte. ; LABORATORY OUTLINES FOR GENERAL BOTANY. 35 XXXVIII. Puccinia graminis Pers. Wheat Rust. Class, Teliosporee. Order, Uredinales. Family, Pucciniacez. The aecial stage of the wheat rust occurs in the spring on the leaves of Berberis vulgaris; the uredinial stage, known as red rust, and the telial stage, known as black rust, occur on the wheat plant. The infected leaves of the bar- berry may be preserved in 70 per cent. alcohol and the wheat leaves and stems may be dried or also preserved in alcohol. : Aecium. 1. Study the under side of a barberry leaf containing the rust under dissect- ing microscope. Sketch an entire left, representing the position of the diseased spots. Fic. 6.— Lire History or PUCCINIA GRAMINIS. 1. Teliospore with basidium (promycelium). 2. Basidiospore (Sporidium,) uninucleate. 3. Barberry leaf with rust spots. 4. Aecium (clustercup). 5. Pycnium (spermogonium). 6. Conidia (spermatia) from the pycnium. 7. Aeciospore (binucleate). 8. Stalk of wheat showing rust pustules. 9. Mycelium from which the uredospores and teliospores are devel- oped. 10. Uredinium with uredospores. 11. Uredospore. 12. Stalk of wheat showing tust pustules. 13. Telium with teliospores. 14. Teliospore. The aeciospore (1.), Uredospore (II.), and teliospore (III.) drawn on same scale. 2. Under low power draw a spot showing the excia—cup-like bodies con- taining the exciospores. 3. Under low power examine a spot on the upper side of the leaf and note the little crater-like openings, which are the necks of hollow bodies, called pycnia (spermogonia), containing thread-like hyphe. Draw. 36 LABORATORY. OUTLINES FOR ‘GENERAL BOD AND eS 4. By means of strips of carrot and a razor cut cross sections of the leaf, mount, and study under low power. Under high power draw an ecium, showing the zciospores. How are they developed? Draw a pycnium with conidia. Uredimium. 5. Under low power, study the diseased spots on the leaves and stems of wheat (Triticum aestivum L.). Draw a patch, showing how the spores break thru the epidemis. 6. Pick out some uredospores with a needle, or if fresh material is at hand cut cross sections of the stem, mount, and draw the uredospores under high power. The uredospores are repeating spores. Telium. 7. Under low power draw a piece of wheat stem containing the black patches of teliospores. 8. Pick out some of the teliospores, or cut cross sections of the stem, mount, and draw a number of spores under high power. Note that the spore is made up of two cells. Study variation in individual spores. Basidwm stage. 9. It is difficult to germinate rust spores in artificial cultures. They germi- nate most readily in spring when those in the field are germinating. ‘Germinate teliospores in a drop culture and study the development of the promycelium (basidium) bearing basidiospores. Draw and describe. 10. Describe in detail the mode of growth and life history of this rust, noting especially the presence of heterecism. XXXIX. (a) FoOmes applanatus (Pers.). (Elfvingia megaloma (Lev.). Phylum, Mycophyta. Class, Basidiomycete. Order, Agaricales. Family, Poly- poracee. This fungus is common on logs and stumps, forming semi-circular brackets or shelf-like bodies from a few inches to a foot or more in width. It is of a wrayish-brown or white color and may be collected at any season of the year. l. Draw the entire fruiting body and describe. The vegetative mycelium is in the wood from which the fruiting body projects. 2. Under low power study a patch of the pores on the under side, by simply laying the fungus on the stage of the microscope and focusing properly. Draw. 3. From a fresh specimen cut cross sections of a piece of the pore-bearing layer, mount, and study the basidia projecting into the cavity of the pores. How many spores on each bascidium. Draw a single spore. 4. Mount some of the brown, woody mycelium from the upper part of the fruiting body. Draw and describe the structure of the fungus. 5. Is the plant a parasite or a saprophyte? Notice the position of the hymenium (pore-bearing surface) in relation to the surface of the earth. Is the mycelium of the fruiting body irritable to the force of gravity? Is there any advantage in this? (b) Polystictus cinnabarinus (Jacq.). Family, Polyporacez. This bracket fungus is very common on dry decaying logs and branches and is easily recognized by its bright red color, especialiy prominent on the under side. 1. Make a sketch of the entire fruiting body. WABORAEROK VS OU TEINES FOR GENERAL BOTANY. 37 2. Under low power draw a patch of the lower surface showing the pores. Note especially the bright red color and compare it with the red color present in many flowers, fruits and roots. How do you explain the presence of the color? XL. Agaricus campéstris (L.). (Psalliota). Common Meadow Mush- room. Class, Basidiomycete. Order, Agaricales. Family, Agaricacez. This edible toadstool grows in open, grassy places in fields and rich pastures. The so-called “bricks” of “spawn” can be obtained from seedmen and will keep for some time when in a dry condition. It can be cultivated by making beds of the proper character in a cellar or greenhouse, or in the open air in gardens. The fruiting bodies may be peserved in 70) per cent. alcohol. 1. Take some of the white filaments or strands from the ground in which the fungus is growing or from a brick of spawn, tease it out with needles and mount in water. Examine under low and high power. Note the numerous hyphe of the mycelium and draw. This is part of the vegetative mycelium which takes up the nourishment from decaying substances in the soil. 2. Examine “button mushrooms” of various sizes and make a series of naked eye sketches showing how the button devolops into the mature fruiting body or toadstool. 3. Study and sketch the mature fruiting body, showing the cap or pileus with gills on the under side, and the stalk with the annulus. Note the irregular fringe at the margin of the pileus. 4. Find the origin of the annulus and the fringe at the margin of the pileus by studying the veil or vellum of a fruiting body in which the pileus is just beginning to expand. 5. Cut off the p:leus of a mature fruiting body and place it gills downward on a piece of white paper. In this way a spore print may be obtained in a few hours. Sketch the spore print. 6. Mount some of the spores and draw under high power. Color and shape? 7. Carefully cut cross sections of the gills of a pileus in which the spores are not quite mature. Mount and study under high power. Draw a part of the hymenial layer (spore-bearing layer), showing the paraphyses and the larger basidia, each of which bears two spores. XLI. Bovista plumbea Pers. Class, Basidiomycete. Order, Lycoperdales. Family, Lycoperdacee. ‘This puffball of a dark-brown color, when mature, is usually abundant in pastures, where it may be gathered in any season. It has a more or less spherical body, usually from one-half to one inch in diameter. 1. Sketch one of the fruiting bodies, showing the inner peridium with an aperture at the apex for the discharge of the spores. 2. Pick out some of the internal mycelium (capillitium) and after moistening with alcohol mount in water. Under high power draw some of the dichotomously branched mycelium and some of the spores. Describe. How does this plant obtain its nourishment? XLII. Lichens. Certain fungi enter into a peculiar parasitic relation- ship with various species of aerial blue-green and green alge. These lichen- forming fungi are mostly Ascomycetae belonging to the phylum Mycophyta. 88 LABORATORY OUTLINES FOR: GENERAL BOTANY. Lichens grow on the bark of trees, on wooden fences, on rocks and on the ground. The common forms may be collected at any time and kept indefinitely in a dry condition in wooden or paper boxes. The peculiar structures called lichens are then simply associations of fungi and algae representing a condition of symbiésis known as helotism; 1. e. the lichen fungus is a slaveholder, the lichen algae are slaves. In the past, lichens were supposed to be an independent group of plants but the lichen-forming fungi will have to be distributed among the proper orders and families of the Mycophyta and the algae among the Schizophyta and Gon- idiophyta. (a) Parmélia cylisphora (Ach.). (P caperdia (L.) ). Subclass, Discoli- chenes. ‘Order, Cyclocarpales. Family, Parmeliacee. This lichen is of a light green color and is very abundant on trees and fence boards and rails, forming large circular thalli often a number oxjamedess im diameter. 1. Study the naked eye characters of the thallus. Draw a part of the thallus, showing the margin. 2. Soak the thallus in water and tease. out a small piece on the slide with needles. Study under high power. Notice two kinds of cells, colorless septate hyphe, the lichen fungus, and green spherical cells, the lichen alge. Draw a piece of the mycelium and some of the alge. To what group do the alge belong? How are the alge and the fungus hyphe arrange in the lichen thallus? 3. Draw two or three alge, showing the manner in which the fungus grows around the green cell to obtain its food. 4. Vegetative propagation. The alga and the fungus each reproduces itself in the manner peculiar to its species, but the lichen may also propagate itself directly by means of little granular flakes produced on the upper surface of the lichen thallus, known as soredia. Mount some of the granular material in water and examine under low power; notice in favorable speciments that the fungus and alge are both present in the soredium. Draw and describe. (b) Lobaria amplissima (Scop.). (Sticta). Subclass, Discolichenes. Order, Cyclocarpales. Family, Stictacez. This is a foliaceous lichen of a light gray color which grows on the bark of trees in forests. 1. Soak the thallus in water and note the change in color of the upper sur- face. Make a sketch showing the position of the brown disk-shaped or cup-shaped apothecia. 2. With a razor, cut free hand cross sections of a piece of the thallus con- taining an apothecium. Hold the piece between two strips of carrot. Mount the sections in water and under low power draw, showing the green algal layer, the white layer and the position of the apothecium. 3. Under high power study the hymenial layer of the apothecium. Draw one of the asci containing spores. Describe. How many spores? Draw a single spore. Draw one of the paraphyses. (c) Dermatocarpon miniatum (L.). (Endocarpon). Subclass, Pyrenoli- chenes. Order Pyrenulales. Family Dermatocarpacee. This lichen with a rather leathery thallus is common on limestone, where it may be obtained at any time of the year. 1. Lay the thallus on the slide without a cover-glass and examine under low power. Draw a part of the thallus, showing the pores which open into the peri- thecia below. LABORATORY OUTLINES FOR GENERAL BOTANY. 39 2. Cut cross sections of the thallus and mount in water. Sketch under low power, showing the algal and fungal layers and the perithecia. 3. Draw one of the perithecia under high power; also an ascus containing spores. (d) Cladonia rangiferina (L.). Reindeer Lichen. Subclass, Discolichenes. Order, Cyclocarpales. Family, Cladoniaceze. This lichen grows on the ground and is generally present on high wooded hills or slopes where it often forms large masses. 1. Sketch and describe a large specimen. 2. Draw a branch showing the apothecia on the branchlets. (e) Collema nigrescens (Leers). Subclass, Discolichenes. Order, Cyclocarpales. Family, Collemacez. A widely distributued, dark colored lichen growing on the bark of trees and on moss. 1. Sketch the foliaceous body, showing the small circular apothecia. 2. Moisten a small piece of the lichen, tease it out with needles, mount and note the chains of Nostoc cells and the fungus hyphe. Draw. 3. Tease out some of the asci or cut sections, mount and draw under high power. How many spores in an ascus? How many cells in an ascospore? 4. Tease out a part of a young lobe of the thallus and also cut sections; mount and examine for slender hyphe which are coiled at the base. These are the so-called carpogonia, the slender projecting part being the trichogyne. In the sections spermogonia, small hollow bodies, may also be found. These bear spermatia on the hyphal branches lining hte walls of the cavity. Draw and describe. THREE INTERESTING GREEN ALG. XLIII. Oedogonium crispum (Hass.). Phylum, Gonidiophyta. Class, Confervere. Order, Oedogoniales. Family, Oedogoniacez. This plant grows either upon or beneath the surface of ponds and pools, usually attached to various solid objects. It fruits most abundantly during May and June, and will grow well in aquaria. 1. Mount some of the filaments in water and examine under low power. Note that the filaments are unbranched and have a definite holdfast at the base. Draw. 2. Draw one of the cells under high power, showing the chloroplast with pyrenoids and the nucleus. Draw the basal cell (holdfast), 3. Nonsexual spore reproduction. If the filaments are in proper condition, any cell may develop into a zoospore and escape from the cell wall. Draw an empty cell. Draw a free-swimming zo6spore and escape from the cell wall. Draw an empty cell. Draw a free-swimming zodspore. These have a circle of short flagella or cilia. Draw a zodspere which has settled down and enlarged and is developing a holdfast at the base. 4. Sexual reproduction. Note the ovaries or oogonia, large cells each with an oosphere filled with food material. Draw. Find the opening at the base for the entrance of the spermatozoid. Draw a spermary or antheridium, usually con- sisting of two or three very short cells each of which gives rise to two sper- matozoids. 5. Look for escaping spermatozoids and for spermatozoids which have en- tered the oogonium. 6. Draw an oogonium containing a ripe, thick-walled oospore. 40 LABORATORY OUTLINES FOR GENERAL BOiAnae 7. When the oospore germinates it divides into four cells, each of which develops into a zoospore. ‘The zoospores settle down and develop into new Oedogonium plants. An attempt should be made to have oospores germinate in a dish of water so that the above mentioned process may be studied. If this stage is at hand draw and describe carefully. 8. Write out the life cycle of this plant in the notes, giving the stages in proper order and noting that the plant has two definite stages in its history — the gametophyte and the sporophyte. XLIV. Ocedogonium borisianum (LeCl.). Family, Oedogoniacee. This species grows in stagnant brooks or in ponds and ditches, usually attached to solid objects. The plants are coarse unbranched filaments and they may be grown in an aquarium. 1. Mount in water and study under low power. Note the long unbranched filament and the basal cell expanded into a holdfast. Under high power draw the tip of a filament, several intermediate cells, and the holdfast. At the summit of certain cells broad zones with peculiar .ring-like striations may be seen. This is where cell division has taken. place. Draw. 2. Draw a single cell, showing a chloroplast with pyrenoids and the nucleus. 3. Study vegetative cells in which the zoospores are developing. These can be seen especially in the morning or in material which has been chilled over night. Draw a single, large zoospore with a circle of short flagella or cilia around the hyaline anterior end. 4. Draw part of a filament, showing ovaries or oogonia, some with an oosphere and an opening in the wall for the entrance of the spermatozoid, and others with thick-walled oospores. Note one or more dwarf males attached to the cell below the ovary. 5. Draw some of the so-called andro-sporangia,; which consist of two to five short cells. These cells give rise to zoospores known as androspores, which settle down on the cells below the oogonia and devolop into dwarf male plants. 6. Draw part of a filament with ovaries or o0gonia and one or more mature dwarf males. Note the large basal cell of the dwarf male and the more slender spermary or antheridium composed of a number of cells. 7. When the oospore germinates after a period of rest the cell breaks out in a delicate sack and divides into a four-celled body (the sporophyte) which gives rise ot four nonsexual zoospores. If material is at hand, study and draw. 8. Write out a careful description of the entire life history of this plant, noting especially that it has an alternation of generations. XLV. Coleochaete pulvinata A. Br. Class, Conferveze. Order, Coleochetales. Family, Coleochetacee. Several species of Coleochete are to be found growing attached to the sur- face of various submerged, fresh water plants. The species mentioned above forms hemispherical masses of closely packed, branched filaments. These masses are large enough to be seen with the naked eye and should be looked for on the petioles or laminze of water lilies and other hydrophytes. 1. Pick off some of the smaller and larger masses with a scalpel, mount, and examine under low power. Under high power draw a part of the branching filaments, showing the joint-like cells, each with a nucleus, chloroplast, and pyre- noid, and some with long ,narrow, hair-like projections sheathed at the base. BABOKATOKRY, QOULLINES HOR GENERAL BOTANY. 4l 2. Look for nonsexual reproduction by means of zoospores, a single one being produced in a cell. If these are present draw and find how they escape from the cell. 3. Draw the mature ovary or oogonium, showing the oosphere and long slender, open neck. How different from Batrachospermum ? 4. Draw one of the spermaries or antheridia, which are terminal or lateal flask-shaped cells of peculiar form easily distinguished from the vegetative cells. Each antheridium produces a single, biflagellate spermatozoid. Compare with Batrachospermum. 5.Draw a mature spermatozoid either free or in the spermary. 6. Draw an oogonium in which the egg has been fertilized, and around Which branches are developing from the base. Bee SuGih mL Pig. 7.— Lire Cycie or: CoLEOCH ATE: fad (7. Draw an ovary containing a ripe oospore and cortical layer of close- fitting branches. 8. From material gathered in early spring or from prepared slides, study fruiting bodies in which a small sporophyte has developed by the division of the oospore into a number of cells. Note the advance of the sporophyte over that of Oedogonium. How many cells does it contain? 9. Each cell of the small, oval sporophyte develops a zoospore, which after a period of activity settles down and develops into a new Coleochete thallus. It is evident from the above that the entire sporophyte of Coleochete is sporogenous. 10. Make a diagram in the notes, showing the life cycle of Coleochete. See Fig. 7. 11. If material is at hand study the flat, disk-like thallus of Coleochete scutata Bréb. Draw under high power and describe. 12. Notre. — The Coleochetacee are the Algz which are most like the plants of the next higher sub-kingdom. On account of the similarity of the body and the life cycle, the ancestors of the lowest liverworts of the present time are sup- posed to have been plants which, with the exception of th sexual organs, were something like these Alge. It must not be supposed, ohwever, that Coleochete Tepresents an intermediate ancestral stage of the liverworts; for, as will appear from the following study, there are very fundamental differences. e SERIES II— ARCHEGONIATA. SUB-KINGDOM AND PHYLUM, BRYOPHYTA. XLVI. (a) Riccia fluitans L. Phylum; Bryophyta.~ Classyshtemaaec: Order, Marchantiales. Family, Ricciacee. This liverwort has a small, linear, dichotomously branched thallus which grows floating in ponds and ditches. It also grows in wet places upon the ground, some- times in cultivated fields. The plant keeps well along with other hydrophytes in a covered, glass jar of water. 1. Mount a small thallus or frond (gametophyte) in water and examine under dissecting microscope. Make a sketch of the plant and describe. 2. Draw a branch of the thallus under low power, showing the air cavities and cellular structure. Note that this thallus is not made up of branching or inter- woven filaments, but that it is a true solid aggregate. Most of the thallophyes. are either simp:e or complex linear aggregates. 3. The aquatic form of this plant is usually sterile. In order to study the sexual organs and sporophyte to advantage, examine prepared slides of Riccio- carpus. (b) Ricciocarpus natans (L.). Family, Ricciacee. This plant forms a small, obcordate thallus which floats on the surface of ponds and swamps. The individuals are hermaphrodite and develop the reproduc- tive organs in the spring and summer. 1. Sketch the thallus under dissecting microscope. Describe. 2. Under low power draw part of a section from a prepared slide, showing an antheridium (spermary). Draw part of a section, showing the archegonium (ovary) containing the oosphere. . 3. Draw an enlarged archegonium (ovary) with the spherical sporophyte, containing a wall one layer of cells in thickness, with the sporocytes lying free in the interior. 4. From older stage draw spore tetrads and mature spores under high power. 5. If prepared slides are not at hand, cut freehand sections of plants (with the aid of strips of fresh carrot roots) with male and female organs and draw an antheridium (spermary) and an archegonium (ovary) under low power. Also cut sections of a plant containing a sporophyte and some free spores. 6. Compare the sporophyte with that of Coleochzte and note the beginning of sterilization of the tissue of the sporophyte. Compare also with Polysiphonia. The antheridium and archegonium may be compared with the plurilocular spor- angia of Ectocarpus. They are not at all like the sexual organs of the higher green alge. 7. Make a diagram in the notes as shown in Fig. 8, which represents the general life cycle for all plants above the thallophytes. Note especially that the diagram represents a life cycle with a true antithetic alternation of generations. (42) / LABORATORY “OUTLINES FOR, GENERAL BOTANY. 43 XLVII. Marchantia polymorpha L. Class, Hepatice. Order, Marchantiales. Family, Marchantiacez. This thalloid liverwort is common on moist rocks and earth, especially on cliffs and around springs. Marchantia as well as Conocephalus and Lunulariz @ Wor fyipos Fig. 8.— DIAGRAM SHOWING PRINCIPAL STAGES IN THE LIFE CYCLE OF THE HIGHER PLANTS. ; can be kept without any trouble in a greenhouse or window garden, provided, they are supplied with sufficient moisture and shaded from intense light by a curtain. Material may be preserved in 70 per cent. alcohol or in the copper salt solution. Gam-tophyte. .. Take a thallus (frond) and notice its dorsiventral position on the ground: Make a naked eye sketch, showing the dichotomous branching, the central groove, and the emarginate growing points. Describe. How is it fastened to the ground? How does the thallus continue its development? How is vegetative propagation accomplished? 2. Under dissecting microscope study the upper surface. Notice that it is mapped off into diamond-shaped areas (areole), each with a small opening in the center (air passage). Draw a patch of the surface. 3. Study the upper surface under low power without a cover glass, by simply laying the thallus on the slide. Draw several areole carefully. The areole represent compartments or cavities in the upper surface of the thallus. The thallus should be kept moist on the under surface as it withers very rapidly. 4. Notice the numerous rhizoids on the under surface. Where are they the most numerous? Mount some in water and draw the three types under high power —one with smooth wall, one with scattered peg-like projections in the interior, and one with somewhat spirally-arranged projections. Of how many cells does each rhizoid consist? Of what use are the rhizoids? Za 44 LABORATORY OQUEEINES \FOR* GEN BAL BOTANY. 5. With a scalpel or knife cut off some of the minute ventral scales arranged in two parallel rows on the under side and forming the central ridge. Mount and draw. res a Ltr CULT AS CI enn 8% Fic. 9.— Lirt Cycle or MARCHANTIA. 6. -Some of the thalli will show brood-bud cups. Under dissecting microscope or low power draw one of the cups containing brood-buds (gemmz). This is a special method of vegetative propagation. r (7. Mount some of the brood-buds and draw. Notice the two opposite grow- ing points and the place where the brood-bud was attached to its stalk. 8. Cut cross sections of the thallus thru a cup, with a razor, and under high power study the development of the brood-buds. Describe. 9. Cut cross sections of the thallus and examine under high power. Where is the main part of the chlorophyll? Draw part of a section, showing the walls of the cavities below the areole, the peculiar chimney-like air passages, the short filaments containing the chloroplasts, and the cellular tissue below the cavities. These details can be worked out better from prepared slides, which should be studied if available and the free hand sections used merely. for comparison. 10. Reproductive branches. Draw a plant with an archegoniophore and one with an antheridiophore. Describe both. Is Marchantia ‘hermaphrodite or uni- sexual? 11. Under high power study prepared slides of cross sections ,cut at right angles to the surface of the disk) of the antheridiophore. Draw an antheridium (spermary), showing the wall, stalk, and the numerous minute cubical mother cells in the interior, each of which will produce two spermatids which develop into spermatozoids. About how many cells in each antherdium? How many antheridia in each disk? Do they all develop at the same time? Notice the openings to the pockets in which the antheridia are situated. About how many spermatozoids are there produced in one antheridiophore? 12. In case no prepared slides are available, cut free-hand sections, mount in water, and study and draw the antheridia under low and high power. PA DOM~A NOK + OULLINE SPOR GENERAL, BOTANY. 45 13. From prepared slides study sections of the archegoniophore. Draw an archegonium (ovary), showing the lid cells, the neck, the neck canal, the venter, the oosphere and the incept of the perigynium (incipient perigynium). Draw the venter of an archegonium showing the ventral canal cell and the incipient oosphere. 14. In case no prepared slides are at hand cut sections of the appendaged disk of the archegoniophore, mount in water, and study the archegonia under low and high power. 15. If male Marchantia plants with properly developed antheridiophores are protected for several days in such a manner that no water will come onto the disks containing the antheridia, active spermatozoids may be obtained in the fol- lowing manner: Place a drop of water on the upper surface of the disk and after a short time take it up with a medicine dropper and mount, or squeeze out several disks on a slide and mount in water. Under high power numerous motile spermato- zoids can be seen, each with two flagella. Study their motion for some time, then stain with a small drop of iodin solution and draw. Sporophyte. 16. Carefully pick out a young light-colored sporophyte inclosed in the peri- gynium, an older stalked one which appears green, and a nearly mature, yellow-. colored one from the under side of the archegoniophore, mount in water and draw under low power, showing the sporangium, stalk, and foot. Describe. 17. ‘If fresh material is available, place a mature sporophyte, which has a ruptured sporangium, on a slide without cover-glass and examine. Breathe gently toward the specimen while making observations. Describe. 18. Under high power draw sporocytes hanging together in chains, and spore tetrads from crushed sporangia, also some mature spores. Draw one of the elaters. What is their function? 19. Make a diagram in the notes showing the life cycle of Marchantia. See Fig. 9. 20. EcotocicaL Note. Describe how the air passages and the character of the nonsexual spores show that Marchantia is adapted to an aerial habitat. XLVIII. Other Thalloid Liverworts. (a) Conocéphalus cénicus (L.). Family, Marchantiacee. 1. Study the thallus of Conocephalus and compare in general with Marchantia. Draw. 2. Under dissecting microscope, draw part of the surface showing the areole with air passages. How do they compare in-size with those of Marchantia? 3. Under low power without cover-glass, draw an areola showing the crater- like air passage. Does Conocephalus have any brood-bud cups? (b) Lunularia cruciata (L.). Family, Marchantiacee. 1. Study the thallus of Lunularia‘and compare with Marchantia and Cono- cephalus. Notice especially the numerous semilunar brood-bud cups. 2. Draw a plant under the dissecting microscope, showing several cups. 3. Under low power draw several areole. How many methods of vegetative propagation has Lunularia? Is there much need for sexual and nonsexual spore reproduction? 46 LABORATORY OUTLINES FOR’ GENERALE BOTANY XLIX. Porélla platiphylla L. (Bellincinia). Class, Hepatice. Order, Jungermanniales. Family, Jungermanniacez. This rather large, scaly liverwort is very abundant on the bark of trees. It may be kept for a long time in good condition in a paper. box. Gametophyte. 1. Moisten a branch of the frond in water and sketch from the upper or dorsal side under the dissecting microscope, showing the arrangement of the Jateral scales. 2. Pick off some scales, being careful so as not to tear off the small, lower, ligulate lobe which may be seen under the large upper lobe of the scale. Draw under low power, showing both lobes of the scale. How many cells in thickness is the scale? Is there any midrib? Why is this scale not homologous with the leaf of a fern or one of the higher plants? The scales are partly analogous to leaves. 3. Draw a few cells under high power. Of what advantage are the thick walls 4. Examine the lower or ventral side of a branch under dissecting micro- scope and note the semicircular ventral scales. Look for rhizoids. Mount one of the ventral scales and draw under low power. Sporophyte. 5. Examine a frond containing little yellowish, club-shaped bodies. These are the sporophytes. Carefully pick out one which has the sporangium unbroken and one which shows the wall of the sporangium split into four valves. Mount in water and draw both under low power, showing the sporangium, the foot and the stalk. : 6. Draw a spore and an elater under high power. 7. Compare the thallus of Porella with that of Marchantia, noting especially the different ways in which the two thalli have been specialized for the work of photosynthesis. L. Sphagnum cymbifolium (Ehrb.). Class, Sphagnee. Order, Sphagnales. Family, Sphagnacee. The peat or bog mosses grow in and near water in swamps, bogs, and other wet places. The species named above is unisexual, the male plant being more slender than the female. Collect plants with sexual organs in winter and early spring and sporophytes in spring and summer. Gametophyte. 1. Take a small mass of dry sphagnum, soak it in water and notice the enor- mous quantity it will absorb. 2. Make a sketch of a female frond. Notice that the frond keeps growing at the top and dying below. 3. Sketch a branch under low power, showing the arrangement of the scales. Draw a single scale. Is there any costa (midrib)? 4. Under high power draw a patch of cells from a scale, some with chloro- phyll and some showing the peculiar spiral and ring-shaped thickenings on the inner surface of the wall. TAO AO A OUMEINES “POR GENERAL BOTANY. 47 §. Mount a piece of the main stem in water and examine under low power. Draw, showing a central brown cylinder and a cortical layer of clear, ee cells with spiral thickenings. 6. Cut off some of the clavate branches at the tip of the male plant, mount, and sketch under low power. Pull off the scales carefully, mount, and examine the antheridia (spermaries). Draw an antheridium under high power. 7. From a female plant carefully cut out an enlarged archegonium (ovary) containing a young sporophyte. Mount and draw under low power, showing the neck at the summit. Around the base some small archegonia may usually be seen. Draw one of these, showing the stalk, venter, neck and lid cells. Sporophyte. 8. Pick out a young sporophyte showing the spherical sporangium, the very short stalk and the expanded bulbous foot. 9. Cut off one of the slender pseudopodia containing a nearly mature sporo- phyte. Sketch under low power showing the sporophyte with sporangium and operculum, and the expansion at the top of the pseudopodium into which the foot fits. 10. Draw some of the non-sexual:spores under high power. 11. From prepared slides make a drawing of a longitudinal section of the sporophyte, showing all the details of the structure. 12. Study and draw an apical cell from a branch of the gametophyte, from a prepared slide. LI. Mosses, General Study. Class, Musci. Order, Bryales. (a) The juvenile gametophyte. When the nonsexual spore of a moss germinates it does not give rise directly to the mature scaly gametophyte, but develops a green filamentous pro-embryo known as the protonema. The protonema can always be found in connection with the very young moss plants which are usually present in greenhouses. The proto- nema may also be found by examining some of the black earth from a place where mosses are growing. The ripe spores of any common species of moss may be sown on moist soil in a box. In a few days, if the box has been covered with a pane of glass, an abundance of green filaments will begin to appear. 1. Place a little earth with young moss plants into a watch glass and care- fully wash off the soil by means of the medicine dropper and needle. Mount the plantlets and any minute masses of filaments present. Examine under low power. Find a good protonema and draw. Notice the branching, the shape of the cells, and the chloroplasts. The similar brown filaments present are rhizoids. 2. Draw a single cell, showing the wall, the cytoplasm, and the chloroplasts. Notice the oblique walls which may be seen in the older filaments. Where and how do the branches originate? 3. Find a protonema which has developed one or more solid green buds from which the mature sexual moss plants will develop. Draw. 4. With what kind of plants previously studied does the protonema com- pare? What then could you call the protonemal stage? How can this be used to explain the evolution of a moss as to habitat, form, and structure? Explain its evolution on this basis; remembering that the protonema is (1) a single cell, (2) a simple filament. (3) a branched filament; and that (4) it finally develops solid buds. These four stages represent the four successive steps in the evolu- tion of the plant body in going from the lowest unicellular forms to the liver- 2 48 LABORATORY OUTLINES FOR GENERAL BO LANG worts. Ontogeny is supposed to partly explain phylogeny. Learn the following law: The history of the development of the individual is an abbreviated history of the development of the race to which it belongs. (b) The young scaly moss plant. Physcomitrium turbinatum (Mx.) (nearly always abundant in greenhouses, by roadsides, and in old fields) or a species of Mnium will be suitable. 1. Mount in water and sketch the entire frond under low power, showing ehe stem, scales, and rhizoids. 2. Draw a single scale, carefully showing the costa and the margin. How does it differ from the scale of Porella? Under high power draw a cell showing the large chloroplasts and thick wall. As in the liverworts these scales are not homologous with true leaves. 3. Draw a branch of a rhizoid. How do these rhizoids differ from those of Marchantia? What relation is there between the rhizoids and protonema? LI. Polytrichum commune L. Common Hair-cap Moss. Class, Musci. Order, Polytrichales. Family, Polytrichacez. The common hair-cap is a widely distributed moss which grows on the ground in old fields and meadows, on hillsides and in woods. The plants are well pre- served in a fruit jar with 70 per cent. alcohol, and collections should be made at various times from winter until early summer when the sporangia are mature. The plants are unisexual and the material for study should include mature male and female plants, female plants with the embryo sporophyte developed just far enough to rupture the calyptra, and female plants with mature or nearly mature sporophytes. Gametophyte. 1. Draw the male and female plants (fronds) of the gametophyte genera- tion, showing the rhizoids, scales, and tip. If the plants are dry or taken from alcohol, moisten in water. Note the rosette of red scales at the tip of the male branch and also the slender green scales at the tip of the female branch. Why this great difference between male and female? 2. Take the tip of a mature male branch and dissect it with needles in a watch-glass, mount the detached parts and examine under low power. Notice the paraphyses and the white club-shaped antheridia (spermaries). Do not mis- take spatulate paraphyses for antheridia) Draw an antheridium under high power. Draw a spermatozoid from a ripe antheridium. 3. Study the living spermatozoids. These may be obtained if suitable male branches are collected after several days of dry weather. Take one of the branches and squeeze out the antheridia onto a slide. Mount in water and ob- serve the motile spermatozoids. 4. If material is at hand, study and draw antheridia from a stained perma- nent mount. 5. Dissect the tip of a female plant, mount the detached parts from the center, and examine under low power. Draw an archegonium (ovary) under high power, showing lid-cells, venter, and stalk. In good specimens the oosphere may be seen. 6. If convenient, study prepared slides containing archegonia. Draw, show- ing the stalk, venter, oosphere, neck canal, and lid-cells. LABORATORY OUTLINES FOR GENERAL BOTANY. 49 7. Cut cross sections of the scaly stem of a large specimen (using pieces of «arrot and razor), mount in water and examine under high power. Draw, repre- senting the epidermal layer, band of peripheral sclerenchyma, inner cortical layer of thinner-walled cells, and central strand. Sporophyte. 8. Select a female plant with a young sporophyte, pull off the calyptra and then pull out the young sporophyte, being careful not to tear off its foot. Sketch the calyptra under the dissecting microscope. What does the calyptra represent? ‘Sketch the young sporophyte under dissecting microscope, showing three regions— foot, short stem, and tip. Remember that the mature sporophyte of Marchantia thas three parts. 9. Draw a mature sporophyte of Polytricum, showing the foot, the stalk or seta, the hypophysis, and the sporangium or capsule. What important advance has the sporophyte of Polytrichum and other mosses made over those of Mar- Ore Game © Fic. 10.— Lire Cycie or PoLytricHuM. chantia and Sphagnum? Note that the sporophyte is a parasite during its entire life. Does it manufacture any food for itself? 10. Cut cross sections of the seta, mount, and draw under high power, show- ‘ing the epidermal layer, the band of sclerenchyma, the layer of thin-walled paren- chyma, and the central strand. The central strand of the sporophyte may be compared with the vascular bundle system of higher plants. 11. Ecological note. The hair-cap mosses are subject to great extremes of ‘moisture and dryness. Let a gametophyte dry out and then place in water. Waht occurs? What adaptation has Polytricum for checking the rapidity of evapora- tion? 12. Make a diagram in the notes showing the life cycle of Polytrichum (see Fig. 10). ~ 50 LABORATORY -OUTLINES FOR GENERALE (OTC ae LIII. Other Mosses. (a) Hypnum radicale Beauv. (Amblystégimm varium (Hedw.). Order, Hypnales. Family, Hypnacez. This is a common moss on decaying logs, in moist, shady places, and on wet ground. Preserve in 70 per cent. alcohol. The sporophytes are mature in summer and autumn. : Sporophyte. | 1. Take a nearly mature sporophyte, lay it on a slide and examine without cover-glass under low power. Pick off the calyptra if still attached and the operculum, being careful so as not to injure the delicate teeth of the peristome. Draw the operculum and the calyptra. 2. Study the hygroscopic movements of the teeth of the peristome by gently breathing on the slide while making observations. Describe the movements. Of what use? | 3 3. Study the true stomata on the hypophysis. Make a sketch of the sporan- gium, showing the peristome and the hypophysis with stomata. The hypophysis may be compared with a leaf of the higher~plants. 5 4. Cut open a sporangium longitudinally and mount. Examine under high power and draw a stoma, showing the two guard cells and some of the surround- ing cells. Draw several of the teeth of the peristome and some of the non-sexual spores. How many teeth are there? Are they in one or two circles? - 5. Cut a cross section of a green sporangium, mount and examine under low power. Sketch the section, noting the following structures: epidermis, hypo- dermal parenchyma, air space, spore sac, central columella. (b) AulacOmnium palustre (L.). Order, Bryales. Family, Aulacomniacee. This moss is common in boggy ground and may be found on charred logs and stumps or on the ground. Collect the material and grow in a moist chamber. 1. Under dissecting microscope make a sketch of a stem, modified in the upper art, the scales of which are easily detached. These scales act as brood buds, and when they fall to the ground are able to develop a protonema. 2. Mount some of the detached scales from near the tip and draw under low power. (c) Leptobryum pyriforme (L.). Long-necked Bryum. Family, Bryacez. | This interesting little moss may be found on moist, shaded cliffs and on rocks near water. It is often very abundant and is easily cultivated in greenhouses, where the gametophyte may be obtained at any season. The rhizoids contain peculiar tuber-like buds of a dark-brown color. On the young sterile fronds these tuber-like bodies are often numerous, being produced on short rhizoids which come from the axils of the scales. 1. Crush the enlarged tip of a plant on the slide, mount in water and study under low and under high power. Draw an antheridum (spermary) and an arche- gonium (ovary); also one of the paraphyses. Note that this gametophyte is her- maphrodite. Compare with Polytrichum. LABORATORY OUTLINES. FOR GENERAL BOTANY. 51 2. Mount and draw a mature brood-bud under high power, showing the structure. Describe. 3. Draw a rhizoid with an enlarged, light-colored end cell, the incept of the ‘brood-bud. mao Y OF FORMS WHICH FORESHADOW SOME OF THE STRUC- TURES DEVELOPED IN THE FOLLOWING SUB-KINGDOM. LIV. Splachnum ampullaceum L. Class, Musci. Order, Bryales. Family, Splachnacee. Altho this odd looking moss is not very common, an attempt should be made to obtain fresh or preserved material of specimens containing mature or nearly ‘mature sporophytes. The plant grows on decaying animal tissue or excreta and is said to occur in cranberry swamps from Ohio to New Jersey and northward. 1. Sketch the entire moss, showing the gametophyte and the sporophyte with the sporangium and the very large, pyriform, fleshy hypophysis. Describe. 2. Sketch the capsule and hypophysis under low power, carefully represent- ing the shape and surface details. 3. Examine the surface of the hypophysis under low and high power and mote the stomata. Draw a small portion of the surface, showing stomata. 4. Nore.—The large hypophysis covered with stomata and filled with loose tissue is well fitted for the work of photosynthesis and may be looked upon as foreshadowing the leaf structure found in the ferns and other higher plants. LV. Splachnum luteum L. or S. rtbrum L. These remarkable mosses are rather uncommon, and very few will probably be able to collect specimens; nevertheless an effort should be made to obtain fresh or preserved material of plants with nearly mature sporophytes. They are reported mainly from the Rocky Mountain region. 1. Under low power make a careful drawing of the sporophyte, showing the foot, the seta, the large umbrella-like thypophysis and the sporangium (Fig. 11). _ 2. Draw part of the surface of the hypophysis under high power, showing the stomata. Are the stomata both on the upper and lower sides? 3. Compare the hypophyses of Hypnum radicale, Polytrichum commune, splachnum ampullaceum, and Splachnum lutewm and note the progressive development of the hypophysis as represented by these types. From this comparison it appears that the hypophysis may be regarded as a nascent, transpiratory and food manufacturing organ. Fig. 11. 2? e q HY LVI. Anthéceros laevis L. or A. punctatus L. SPOROPHYTE OF SPLACH- Class, Anthocerotee. Order, Anthocerotales. Family Anthocero- NUM tacee. LUTEUM. The hornworts are common on wet banks and sandstone ledges, especially around springs and shady places. The gametophyte is a small, lobed, more or less 52 LABORATORY .OUTLINES FOR GENERAL BOTAN. disk-shaped thallus from which the sporophytes.extend upward like small ver- tical horns. Preserve in copper salt solution. 1. Under dissecting microscope, sketch a gametophyte containing nearly mature sporophytes. Note the thick tubular sheath around the base of the sporophyte. 2. Mount a small piece of the thallus and under high power draw a cell showing the single large chloroplast. Compare these cells with those of Coleo- chete. 3. Look for endophytic colonies of a blue-green alga (Nostoc) in cavities on the under side of the thallus. 4. Separate a sporophyte, which is just mature at the tip, from the gameto- phyte, being careful to keep the foot in a perfect condition, and sketch under low power. Represent the slender sporangium, the bulbous foot with wart-like out- growths, and the short stalk with a growing zone between the foot and sporangium proper. Under high power note the stomata in the green tissue toward the base of the sporangium. Draw. 5. Study a sporophyte in which the tip of the sporangium has split open. Notice the columella. 6. Mount some of the spores and spore tetrads and draw under high power. Describe the spore tetrads. 7. I£ prepared slides are at hand, the aeons of the foot, the growing region, and the sporangium should be worked out. Note especially the arrangement of the elaters, which have a tendency to separate the cavity of the sporangium into tranverse compartments. 8. Note.— The hornworts come nearer to the lowest ferns than any other Bryophytes and it is probable that the Bryophyte ancestors of the lowest Pterido- phytes were something lke a horned liverwort with perhaps a chlorophyll-bear- ing tissue arranged somewhat like the hypophysis of a Splachnum. Anthoceros also points to the Pteridophytes in that it has the sexual organs embedded in the thallus. In Splachnum and Anthoceros together appear five structures which fore- shadow or anticipate important structures in the Pteridophytes. These are: (1) the bulbous foot and wart-like outgrowths of Anthoceros; (2) The central strand in the seta of Polytrichum and other mosses; (3) the intermediate growing zone at the base of the Anthoceros sporangium; (4) the large hypophysis of Splachnum with numerous stomata; and (5) the arrangement of the spores and elaters (sterile tissue) in the sporangium of Anthoceros. SUBKINGDOM,. PTERIDOPHYTA HOMOSPOK A SERIES OF FERNS TO TELUSTRADE i EVO Uae OF COM- PLEX ORGANS FROM SIMPLE ONES: LVII. Ophiogl6ssum vulgatum L. Adder-tongue. Phylum, Ptenophyta. Class, Filices. Order, Ophioglossales. Family, Ophioglossacee. This simple fern is mature about the middle of June and may be found in moist meadows and thickets. The entire plant should be dug up and care taken so as not to injure any of the roots. Preserve in copper salt solution and press some for herbarium specimens. Altho fresh plants are preferable, preserved’ or dry herbarium specimens will answer very well. EABOhALOIRY: OOREINES- hOR “GENERAL BOTANY. 53: Sporophyte. 1. Sketch an entire plant, carefully representing the four important regions of the sporophyte—sporangiophore, leaf blade, stem (short upright rhizome), and roots. Do the roots branch? Note the growing pene at the summit of the rhizome from which new leaves are developed. 2. These four parts may be compared with the organs of a Splachnum. sporophyte in a general way as follows: a. The sporangiophore with the sporangium. b. The leaf-blade with the hypophysis. c. The stem with the seta. d. The root system with the foot. Compare also with Anthoceros, noting especially the growing bud in the stem. 3. Study the venation of the leaf under low power. Draw a-portion and describe. Study and draw the stomata. 4. Mount a part of the nearly mature sporangiophore. Draw under low power, showing the sporangia. Under high power draw some of the nonsexual. spores. LVIII. (a) Botrf¥chium simplex Hitch. Little Grape-fern. Class, Filices. Order, Ophioglossales. Family, Ophioglossacez. This fern is to be found in moist woods and meadows and Sone be gath-. ered about the middle of June. 1. Sketch the entire sporophyte from fresh or herbarium specimen and note the advance in complexity, over Ophioglossum, of the sporangiophore, leaf and roots. (b) Botrychium lunaria (L.). Moonwort. The moonwort is found in the northern part of the United States and usually grows in fields. 1. Sketch the entire sporophyte, showing the sporangiophore, leaf, roots and rhizome. Describe how this plant differs from the preceding. (c) Botrychium negléctum Wood. Matricary Grape-fern. This fern grows in grassy woods and swamps and should be collected about the middle of June. 1. Sketch the entire sporophyte and note the advance in complexity over the moonwort. : LIX. Botrychium obliquum: Muhl. Oblique Grape-fern. This evergreen grape-fern is widely distributed and may be collected in summer and autumn. Good herbarium specimens are satisfactory. 1. Sketch the entire sporophyte and note the advance in complexity over - the preceding in the development of the sporangiophore, leaf, and roots. 2. Study th venation of the leaf under low power. Is there any relation between the development of lobes and the venation? 3. Mount a branch of the sporangiophore and draw several sporangia under low power. Draw some of the non-sexual spores. 4. Ecological note. Notice the strong root-contraction and draw under dis- secting microscope. How does the upright rhizome which continues to grow upward keep in the ground? 54 LABORATORY OUTLINES FOR \GENERAL BOIANY LX. Botrychium virginianum (L.). Virginia Grape-fern. The Virginia grape-fern is common in rich woods and should be collected in the summer. Good herbarium specimens are satisfactory. 1. Sketch the entire sporophyte, showing especially the extreme complexity of the leaf. What relation is there between the ultimate segments of the leaflets and the venation? 2. Compare the last five plants in regard to the sporangiophore, the leaf- blade and the roots. 3. Cut cross sections of the rhizome, mount and draw a sector under low power, showing the wide cortex, the endodermis, the phloem, the cambium, the xylem (wood) with medullary rays, and the central pith. This type of vascular cylinder is called a siphonostele. 4. Cut longitudinal sections of the fleshy root tips, mount the central sec- tions, and draw a tip showing the apical cell. 5. The gametophytes of the grape-ferns are subterranean and difficult to find. They are destitute of chlorophyll and have the appearance of minute tubers. Ii fresh or preserved material of the gametophyte of the Virginia grape-fern is at hand, study and sketch under dissecting microscope or low power, showing the general contour of the body and the rhizoids. 6. Notr.— The advance from such forms as Splachnum and Anthoceros to Ophioglossum represents a vertical evolution, i. e., evolution upwards. The - development indicated in passing thru the series of forms from Ophioglossum vulgatum to Botrychium virginianum represents a horizontal evolution. There is a close relationship among these ferns. It must not be supposed, however, that the latter has necessarily been derived directly from the former, but only that the ancestors of Bothrychium were at one time in a condition as simple as Ophioglossum is at the present time. GENERAL STUDY OF. HOMOSPOROUS PYERIDOEREAA Es: Exe Bers (a) Adiantum capillus véneris L. Venus-hair Fern. Phylum, Ptenophyta. Class, Filices. Order, Filicales. Family, Polypodiacee. The venus-hair fern grows in ravines and is widely distributed, but very rare in the North. It grows very readily in. greenhouses, and is extensively cultivated. The gametophytes may be found at almost any time on pots in greenhouses. They may ‘be raised in large quantities by sowing spores on any well-prepared, moist ground. Gametophyte. 1. Mount a fresh, heart-shaped thallus in water and sketch it from the upper side under dissecting microscope. 2. Study the rhizoids under low power and draw a single one under high power. Are they unicellular or multicellular? 3. Under high power draw a single cell of the thallus, showing the chloro- plasts. 4. Examine the lower side carefully under low power and note the numer- ous antheridia (spermaries) and archegonia (ovaries). How are these organs distributed over the thallus? Under high power draw an antheridium and an archegonium (so much as can be seen of them above the surface of the thallus). Compare the thallus of Adiantum with Anthoceros and Marchantia . Note especially the comparatively small size of the gametophyte and that it is hermaphrodite. PABOkCAnORY SOO REINES HOR IGE NER AL BOTAN Y. 55 ®). Look for the large, spirally coiled spermatozoids moving in a ripe anther- idium. Study free-swimming spermatozoids and draw. Describe the movement. The spermatozoids can usually be found on gametophytes of proper size and are often present in large number. lodin will bring out the flagella. 6. If prepared slides are available, draw a section of a nearly mature anther- idium, showing spermatozoids in various stages of development; also draw an archegonium, showing the neck, venter and oosphere. 7. Study young gametophytes and recently germinated spores. Note the protonema. Draw. Compare with the Bryophytes. ~Sporophyte. 8. Sketch and describe the compound leaf. 9. Mount and draw a single leaflet under dissecting microscope, showing the general outline and the free, dichotomous venation. How does the character of the venation explain the notched and cut margin? Can the origin of the leaflets be explained from the same standpoint? Note that the tips of some of the lobes are bent under so as to cover the sporangia. 10. Under high power study and draw the stomata. Are they on the upper or lower surface or on both? Draw a single cell, showing the chloroplasts. 11. If slides are at hand draw a section of a young sporophyte embryo, showing four definite regions (quadrants). 12. Pick out a young sporophyte from the under side of an old gametophyte, and draw under low power. Show the four regions, first leaf, root, bud and foot. Note that the young sporophyte is parasitic on the parent gametophyte, and that it acquires its independent life gradually. 13. Cut sections of the rhizome, stain and mount or use prepared slides. Note the general ground tissue and several, scattered, concentric vascular bundles. Draw. (b) Pteridium aquilinum (L.). (Pteris). Eagle-fern. Family, Polypodiacez. The eagle-fern grows on hillsides, especially in sunny places. The rhizomes: should be preserved in alcohol. 1. Cut cross sections of the rhizome, mount, and sketch under dissecting microscope, representing the following structures; the band of external or cortical sclerenchyma, the pith or ground tissue, internal sclerenchyma (stereome) in two large, brown bands and in smaller patches, and the concentric fibro-vascular bundles —usually three large ones and a number of smaller ones. Note the two lateral ridges. How do you account for the dorsiventral condition of the rhizome? This type of vascular system is known as a polystele. 2. Under high power, make a careful drawing of one of the smaller vascu- lar bundles, showing the bundle sheath, usually brown, the phloem and the central xylem (wood). 3. Test for starch with iodin solution. Draw some of the ground tissue, showing the intercellular spaces, and starch in the cells. 4. Draw a patch of cells from the internal and from the external scleren- chyma, showing the thick cell walls. 5. Cut longitudinal sections of the rhizome, mount and draw, comparing the structures with those seen in the cross section. Also draw a single cell from the external sclerenchyma, the internal sclerenchyma, and the ground tissue. From the vascular bundle, draw a sieve tube and a scalariform tracheid. 56 LABORATORY OUTLINES: FOR. GENERALE. BORANiE 6. Describe the mode of growth of the Pteridium rhizome. What advantages in the geophilous habit? Has this rhizome any advantage over the vertical rhi- zomes of Ophioglossum and Botrychium? 7. Carefully remove the leaves from the apex of a branch ee the rhizome and cut cross sections of the growing point. Mount the sections, and in the first two or three look for the apical cell. Draw. Cut longitudinal sections of the apex of another branch, mount and draw the section, showing the apical cell. What is the shape of the apical cell? 8. Under dissecting microscope draw a leaflet of Pteridium from the lower side, showing the membranous false indusium formed of the reflexed margin of the leaflet. 9. If fresh, young leaves are at hand, study the nectar glands with drops of nectar in the axils of the main divisions. Locate them and draw. Of what use are the nectaries? (c). Cyrtoémium falcatum J. Sm. Family, Polypodiacez. Cyrtomium grows readily in greenhouses: and window gardens, and fresh sporangia may be obtained at almost any time of. the year. 1. ‘Examine a sterile leaf and a sporophyll. Draw one of the leaflets showing the circular sori on the under side. Fe. 12.— Lire CycLe oF ORDINARY FERN. Pick off some of the sori which have recently exposed the sporangia, examine without cover-glass and describe how the spores are scattered. 3. Mount an indusium and some opened and unopened sporangia. Draw the indusium under low power. 4. Draw a single sporangium under high power, showing the stalk, annulus, and lip cells. Contrast this sporangium with the one in Botrychium. Draw some of the nonsexual spores. Note shape, color and surface. ~ 5. Make a diagram in the notes showing the life cycle of a fern. See Fig. 12. LABORATORY OULEINES FOR GENERAL: BOTANY. 57 LXII. Other Ferns. (a) Camptosorus rhizophyllus (L.). Walking-fern. Family, Polypodiacee. The walking-fern is common on rocks, especially on limestone. Study fresh or herbarium specimens. Sporophyte. . 1. Sketch a plant with several leaves, some of which have rooted at the long acuminate tips, and show plantlets of various sizes. This is a simple and effective method of vegetative propagation. Leaves are usually highly specialized organs which have to a large extent lost the power of reproducing the individual. There are, however, many cases, like the present one, even in the higher plants, where the leaves retain the power of reproduction to a remarkable degree. (b) -Filix bulbifera (L.). (Cystopteris.) Bulbiferous Bladder-fern. Family, Polypodiacee. This fern grows on moist rocks, especially limestone, and is easily cultivated in greenhouses, where it propagates itself extensively. Sporophyte. 1. Sketch a leaf showing a number of fleshy brood-buds (bulblets). On which side of the leaf are they developed? 2. Under dissecting microscope draw a brood-bud which has just fallen off. 3. Draw a young fern plant which is developing from a brood-bud. Is this an efficient method of vegetative propagation? Why? (c) Marattia douglassii (Presl.). Order, Marattiales. Family, Marat- tiacee. This fern may be obtained in large greenhouses and conservatories. Use either fresh or preserved material. 1. Draw a leaflet showing the peculiar sori. 2. Draw a single sorus under low power and describe. This is a eusporan- giate fern. How do the sori and sporangia differ from those of Cyrtomium? LXIII. Lycopdédium lucidulum Mx. Shining Club-moss. Phylum, Lepidophyta. Class, Lycopodiee. Order, Lycopodiales. Family Lycopodiacez. This lycopod grows in moist woods and on shady cliffs. Use fresh, alcoholic, or herbarium material. Sporophyte. 1. Sketch the entire plant. Note the dichotomous branching, the alternating zones of sporophylls and sterile leaves, and the dichotomous roots. 2. Draw a branch, showing very carefully the tip and several zones of sporo- phylls below. Note that the formation of sporophylls does not stop the growth of the axis on which they are produced. Which are the larger, sporophylls or sterile leaves? 3. Draw a single sporophyll with sporangium under low power. 4. Under high power draw several non-sexual spores. 58 LABORATORY OUTLINES FOR GENERAL BOTANY. 5. From alcoholic material cut cross sections of the stem, mount, and draw under lower power. Note the epidermis with cuticle, the wide cortical layer, the vascular bundles of the leaf traces, the bundle sheath or endodermis, and the central cylindrical mass of vascular tissue. Inside of the endodermis are a number of more or less parallel strands of xylem and phloem. These structures will be more prominent after staining with iodin solution. This type of vascular system is called a protostele. 6. Cut radial longitudinal sections of the stem and compare in detail with the cross section. 7. Vegetative propagation. Notice the peculiar bulb-like brood-buds near the tips of some branches. Pick off one and draw under dissecting microscope. LXIV. Lycopodium obscurum L. Tree Club-moss. Lycopodium obscurum grows in moist woods, forming long slender rhizomes which creep under the surface of the ground or under leaf mold. From this rhizome upright, zrial branches develop. Sporophyte. 1. Sketch an entire plant showing the rhizome and upright branch bearing a number of cones. 2. Draw a single cone under dissecting microscope. Note the spiral arrange- ment of the specialized sporophylls, and that by the development of a cone the further development of the axis is stopped. What is the probable reason for this? 3. Under low power draw a single sporophyll showing the sporangium. ~ Note the advance in specialization of this sporophyll over that of the preceding plant. : 4. Under high power draw some of the nonsexual spores; also some of the spore tetrads from younger sporangia. 5. Notr.— This cone represents a primitive flower. Compare it with the zone of sporophylls in the preceding species. LXV. (a) Equisétum arvénse L. Field Horsetail. Phylum, Calamophyta. Class, Equisetee. Order, Equisetales. Family, Equise- tacee. The field horsetail is common along roadsides and railways, on river banks and steep slopes facing the north. The fertile branches come up in April and May, while the sterile ones begin to appear at about the same time, but do not reach their full development until later in the season. Spores may be collected in large quantities and kept in a dry glass bottle. Rhizomes with fertile and sterile branches should be preserved in 70 per cent. alcohol. Good hebarium specimens may also be used. Sporophyte. 1. Sketch a plant containing the rhizome, fertile shoot with cone, and young sterile shoot. Note the whorls of scale-like leaves at the nodes; also the lack of chlorophyll in the fertile shoot. 2. Sketch a mature sterile shoot. 3. Note and describe the division of labor in the stem of this plant — rhi- zome for a food storehouse and for vegetative propagation, fertile branch for the production of non-sexual spores, sterile branch with abundant chlorophyll for food manufacture. From whence is the food material obtained which goes to form LABORATORY OUTLINES FOR: GENERAL-BOTANY. 59 the fertile shoot? Compare the stems of Lycopodium lucidulum, L. obscurum and Equisetum arvense and note the degree of differentiation in each. 4. Cut off some of the peltate sporophylls, mount and draw from the side under dissecting microscope. Show the stalk, the angular outer expansion and the sack-like sporangia hanging from the under side. How are the sporophylls arranged in the cone? Compare this cone and the sporophylls with those of Lyco- podium obscurum. Compare also these two sporophylls with a fern sporophyll. 5. Place a small flake of the dry spores on a slide without water or cover- glass, breathe on them gently until the glass becomes moist, and examine imme- diately under low power. Note the spores with appendages coiled about them. In a few moments the spores will be in violent agitation, while the appendages uncoil. Breathe gently on the slide while looking into the microscope. How many appendages on each spore? Draw. Describe in detail the hygroscopic prop- erties of the appendages. Of what advantage to the plant is this peculiar ar- rangement? 6. Cut cross sections of a young fertile branch from alcoholic material. Mount, stain with iodin solution and draw under low power. Note the epider- mis, the wide cortical layer with a circle of. lysigenous cavities the endodermis, the circle of vascular bundles, and the pith with a large central lysigenous cavity. The xylen (wood) of each vascular bundle is arranged somewhat in the form of a V, the apex of the V being occupied by a large air-cavity. The two limbs of the V end near the endodermis, and the phloem is situated between these two masses of xylem. . (b) Equisétum hyemale L. Scouring Rush. This plant grows in wet places along the banks of rivers, creeks and lakes. 1. Examine the fresh or dry stems under low power. Notice the parallel erooves and ridges, with lines of tubercles and stomata. Draw and describe. 2. Break some of the dry stems and note their brittleness. Burn one of the stems in a hot flame, mount the outer part of the shell which remains, and examine under low power. Notice that the cell walls and stomata are still distinct. This is because the cell walls are impregnated with silica. Draw a flake showing the stomata. . 3. Note.— The cavities often contain water or ice in the winter. SUB-KINGDOM, PTERIDOPHYTA HETEROSPORE-. LXVI. Marsilea quadrifélia L. European Marsilea. Phylum, -Ptenophyta. Class, Hydropteride. Order, Marsileales. Family, Marsileacez. This water fern grows well in artificial ponds, in gardens and greenhouses. The western Marsilea vestita H. & G. found in wet places and shallow ditches on the great plains and prairies of the interior may also be used. The sporophytes are mature in autumn. Sporophyte. 1. Sketch a branch of the creeping rhizome, showing the roots and the leaves with slender upright petioles. 2. Sketch a sporophyll with two sporocarps. 3. Carefully cut off part of the thick inner margin of some sporocarps and place them in a glass of water. In a day or two a gelatinous ring will be ex- 60 LABORATORY OUTLINES: FOR GENERAL BODAN truded on which are situated the sack-like sori in which microsporangia and mega- sporangia are contained. Draw. 4. Mount some of the microsporangia and megasporangia and draw each under low power. The megasporangium contains a single megaspore; the micro- sporangium a considerable number of microspores. 5. Under high power draw a single microspore and megaspore, in correct proportion. Gametophyte. 6. In the meantime the spores will begin to develop the gametophytes. These are very minute, and the spores in the water should be examined every few hours in order to get the proper stages. The male gamtophyte develops entirely in- side of the microspore wall and the female gametophyte merely protrudes the neck of the archegonium (ovary) from one end of the spore. Draw a male gameto- phyte with a protrusion on the side of the spore wall for the escape of the sper- matozoids and a female gametophyte with archegonium projecting from one end, showing a large number of spermatozoids in the gelatinous substance extending from the neck of the archegonium. Why does the microspore always give rise to a male plant, and the megaspore to a female? 7. If prepared slides are at hand, study and draw sections of nature male and female gametophytes. The male gometophytes correspond to the pollen grain of seed plants, and the female gametophyte to the embryo-sac in the ovule. Both gametophytes of Marsilea must be compared with the hermaphrodite gametophyte of Adiantum. Note especially the great reduction in size; also that after this there will be no more hermaphrodite gametophytes, hence no possibility of self- fertilization. 8. In a week or so the female plants in the glass of water will have embryo sporophytes. Draw under low power and describe. 9. EcoLtocicaL Nore—Examine a plant at night, or place a flower-pot with a living plant in a dark chamber and note the manner in which the leaflets fold up. How long does it take the leaflets to unfold after being placed in sunlight? LXVII. Salvinia natans (L.). Salvinia. Class, Hydropteride. Order, Salviniales. Family, Salviniacee. This floating water fern grows readily in aquaria in greenhouses. Sporophyte. 1. Draw an entire plant as it floats on the surface of the water, showing the horizontal stem, the leaves, and the peculiar root-lke leaves hanging down from the underside. 2. Take out some of the plants and throw them into water. Note how they nearly always turn right side up. 3. Place a leaf on'‘the slide and examine without cover-glass under low power. Draw a part of the surface showing the peculiar hairs. What is their use? Mount one of the dissected, root-like leaves and sketch under low power. 5. EcotocicAL Note.—Describe the various ways in which the sporophyte of Salvinia is adapted to its environment. LXVIII. Isdetes melanépoda J. Gay. Black-based Quillwort. Phylum, Ptenophyta. Class, Isoetee. Order, Isoetales. Family, Isoetacaee. This quillwort may be found in moist prairies and overflowed fields in the LABORATORY OUTLINES: FOR: GENERAL BOTANY. 61 central states of the Mississippi Valley. Fresh or herbarium specimens may be ised, and stems preserved in 70 per cent. alcohol. Sporophyte. 1. Sketch and describe the entire sporophyte, showing leaves, short stem, and ‘roots. 2. Study prepared slides or cut cross sections of stems in alcohol and draw, showing the following structures: the two vertical furrows and two large lateral lobes, the outer cortex and extensive parenchymatous tissue in which the cells are arranged in radial rows, on the inner limits of this layer a zone of meristem- atic cells, inside of this a layer of clear cells (the phloem, “prismatic layer”) and ‘in the center a xylem-cylinder from which bundles pass outward to the leaves. LXIX. Sigillaria sp. Phylum, lLepidophyta. Class, Selaginellee. Order, Sigillariales. Family, ‘Sigillariacee. Fossil impressions of the trunks of large, arboreous Sigillarias are common in the formations of the carboniferous period and may be seen in most museums. 1. Sketch the surface of part of a trunk of Sigillaria, showing the leaf scars and the longitudinal fluting. 2. Notre.— The heterosporous pteridophytes of the present time are the rem- nants of a once great group of plants which formed a characteristic vegetation ‘before and during the carboniferous period, which ended millions of years ago. LXX. Selaginélla kraussiana (Kunze). Krauss’ Selaginella. Phylum, Lepidophyta. Class, Selaginellez. Order, Selaginellales. Family, Selaginellacez. This plant grows very luxuriantly in greenhouses and window gardens, if the soil is provided with proper moisture. Suitable material may be had at any time -of the year. Sporophyte. 1. Sketch an entire plant, showing branches, leaves, and roots. Note that the branches occur only in one plane and that the roots are dichotomous. Describe the character and arrangement of the leaves. How do you account for the arrangement? How does the plant accomplish vegetative propagation? 2. Draw a leaf under low power. Under high power draw a cell, with a ‘single chloroplast and one with several chloroplasts. Draw also one of the stomata. Where are the stomata situated? Look for the ligule on the leaf. Of what use is the ligule? 3. Cut cross sections of a fresh stem or of stems preserved in alcohol, ‘mount, and draw, representing the following structures: epidermis, cortical tissue ‘in which may appear sections of bundles passing to the leaves, two or more large air spaces, and in the center of each space a vascular bundle. The bundle consists of a central strand of xylem (wood) surrounded by a band of phloem which is enclosed in a large-celled bundle sheath. Note the strands of cells passing through the air space from the cortex to the vascular bundle. 4. Draw one of the short bisporangiate cones (primitive flower) under ‘dissecting microscope, showing microsporophylls above and one or more mega- sporophylls below. 5. Carefully pick off a microsporophyll and a megasporophyll each with its ‘sporangium, mount and draw under low power. Note the greater specialization 62 LABORATORY OUTLINES FOR GENERAL BOTANY. in the arrangement of the sporangia over that of Marsilea. How many mega-— sporophylls in comparison with the microsporophylls on each cone? Note the numerous microspores in the microsporangium. How many spores in the megasporangium ? 6. Draw a microspore and a megaspore in exact proportion under low power. How do you explain this difference in size of the nonsexual spores? Determine: how many times greater in volume the megaspore is than the microspore. How many megaspores and microspores in one cone? Gametophyte. 7. From prepared slides draw the male and female gametophytes of Selaginella, the archegonium (ovary) and antheridium (spermary) and the oosphere and spermatozoid. Why does the microspore always produce a male Fie, 13) irs Cvcre sory SEEAGINERIUAS and the megaspore a female gametophyte? Compare with Note 12 under Vaucheria. Observe also that the determination of sex in these plants, as well as. all other heterosporous groups, has no relation to the reduction division. 8. Draw and describe a young sporling showing root, stem and first leaves with megaspore still attached containing the female gametophyte and foot of the sporophyte. . 9. Make a diagram in the notes showing the life cycle of Selaginella. See Fig. 13. 10. Nore.—It will be remembered that in the lowest archegoniates the gametophyte is the important plant in the life cycle, and that the sporophyte is very small. Now in the highest forms the tables are turned and the sporophyte has become the plant. Between such plants as Isoetes and Selaginella on the one hand and the lowest living seed plants on the other, there is a considerable hiatus, nevertheless it is not difficult to trace the transformation which was necessary in passing from the condition of heterosporous pteridophytes to the lowest gymmo- sperms. ONE XLV. 2, GUsle XII. XI. DS EEE. DADOKANOKY OUOTEINES FOR GENBKAL BOTANY. 63 PHYA SND CLASSES, OF THE. PLANT KINGDOM. Anthophyta ......{ Dicotylae — Dicotyls. (Flowering Plants) Monocotylae — Monocotyls. Strobilophyta ....{ Gneteae — Joint-firs. (Strobilus Plants) ) Coniferae — Conifers. { Ginkgoeae — Maiden-hair dirzees: | Cordaiteae (Fossil) — Cor- Cycadophyta ..... daites. (Cycad Plants) Cycadeae — Cycads. Pteridospermae (Fossil)— Seed-ferns. Lepidophyta ..... § Selaginelleae — Selaginellas. (Scale-leaf Plants) | Lycopodieae — Lycopods. { Sphenophylleae (Fossil)— Wedge-leaf Calamites. Calamophyta Calamariae (Fossil) — (Calamite Plants) | Calamites. [ Equiseteae — Horsetails. Isoeteae — Quillworts. Ptenophyta ...... 4 Hydropteridae — Water-ferns. (Fernworts) |; Filices — Ferns. ( Anthoceroteae — Hornworts. Musci— True Mosses. Bryophyta .......{ Andreaeae—Granite Mosses. (Mossworts) | Sphagneae — Bogmosses. | Hepaticae — Liverworts. f Basidomycetae — Basidium Fungi. Teliosporeae — Brand Fungi. Mycophyta’ 5.5: .: Laboulbenieae — Beetle Fungi. (True Fungi) | Ascomycetae— Sack Fungi. | Oomycetae. | Zygomycetae. Charophyta ...... { Chareae — Stoneworts. (Stoneworts) ] Rhodophyta ..... { Florideae— Red Seaweeds. (Red Algae) l Bangieae. ( Dictyoteae. Phaeophyta ...... 1 Cyclosporeae — Rockweeds. (Brown Algae) [ Phaeosporeae — Kelps. SUBCLASSES. Dicotylae— Inferae. Tubiflorae. Heteromerae. Myrtiflorae. Amentiferae. Calyciflorae. Centrospermae. Thalamiflorae. Monocotylae. Liltiflorae. Glumiflorae. Spadiciflorae. Helobiae. 64 LABORATORY. OUTLINES FOR GENERAL BOTANY. PHYLA AND CLASSES OF THE PLANT KINGDOM — Concluded. Conferveae — Confervas. Siphoneae — Tube Algae. Monoblepharideae. IV. Gonidiophyta ....} Hydrodictyeae. (Zoospore Plants) ) Archemycetae — Primitive Fungi. Protococceae. Pleurococceae. Lik, eZygophyta- =... { Conjugatae. (Conjugate Algae) ) Diatomeae — Diatoms. Il. Myxophyta .......( Myxomycetae — Slime Molds. (Slime Plants) Plasmodiophoreae. f Myxoschizomycetae — Slime | Bacteria. I. Schizophyta ..... Schizomycetae—Fission Fungi. (Fission oe Glaucocysteae. Cyanophyceae — Blue-green Algae. Hel, SERIES III— SPERMATOPHYTA. SUB-KINGDOM GYMNOSPERME. LXXI. Cycas revoltita L. Cycad. Phylum, Cycadophyta. Class, Cycadee. Order, Cyadales. Family, Cycadacez. This plant is usually grown in greenhouses and conservatories. Herbarium and museum material should also be at hand. 1. Examine a living plant and describe its general character. Sketch the stem, showing the scale leaves and one foliage leaf. 2. Draw a megasporophyll (carpel) from herbarium specimens, showing the megasporangia or ovules. Note the similarity of the carpel to the foliage leaves. The carpels are produced in a whorl like the foliage leaves, and the stem continues to grow thru the whorl. Compare this condition with the ordinary ferns and with Lycopodium lucidulum. 3. Make a sketch of the large staminate (microsporangiate) cone. Draw a single microsporophyll (stamen), showing the numerous microsporangia on the under side. 4. From alcoholic material draw a young ovule, properly dissected, showing the integument with micropyle, the inner wall of the ovule (megasporangial wall) with the pollen chamber, and the female gametophyte. 5. Draw half of a large female gametophyte from a mature seed, showing the little depression at the outer end and the dormant sporophyte embryo. The necks of the archegonia open into this depression (called the archegonial chamber) at the time of fertilization. 6. Mount male gametophytes (pollen-grains), and draw under high power. 7. If prepared slides are available study sections of pollen-grains showing the internal structure. 8. Notre.— The fundamental difference between the heterosporous pterido- phytes and the lower seed plants is that in the latter the microspores and mega- spores are not shed, but develop the male and female gametophytes in the micro- sporangia and megasporangia respectively, while in the former the spores sooner or later drop to the ground. The female gametophyte remains permanently enclosed in the megasporangium, but the male gametophytes are shed from the miscrosporangia and some fall into the micropyle of the ovule. This is known as pollination. In order that the spermatozoids may fertilize the oospheres in the archegonia a short pollen tube must grow thru the tissue between the pollen chamber and the female gametophyte. It will be observed that the gametophytes are now entirely parasitic, the female in the ovule and the male at first in the microsporangium, and after pollination, in the wall of the ovule. LXXII. Ginkgo biloba L. Maiden-hair-tree. Phylum, Cycadophyta. Class, Ginkgoee. Order, Ginkgoales. Family, Gink- goacee. This beautiful tree, a native of China and Japan, is cultivated quite exten- sively in the United States. Museum and herbarium material may be used. 5 (65) 66 LABORATORY OUTLINES FOR GENERAL BOTANY. Sporophyte. 1. Sketch a leafy branch, showing the leaves developed in clusters on dwarf branches. Note that dwarf branches may give rise to ordinary branches. 2. Sketch a single leaf under dissecting microscope, showing the dichotomous venation. Compare the venation with that of the Adiantum. leaf. 3. Sketch a stamen -(microsporophyll) under lower power. How many microsporangia? Compare with stamen of Cycas. Sketch a carpel (megasporo- phyll) and compare with the Cycas carpel. 4, Sketch a mature fleshy seed on its long stalk. Note the collar or cup around the base of the seed and the small undeveloped ovule. On some stalks two seeds develop. Remove the fleshy part of the integument and note the hard, inner layer. : Gametophyte. 5. Draw a male plant (pollen-grain) under high power. 6. From alcoholic material study the mature female gametophyte (kernel of the seed). Sketch, and compare the size of the male and female gametophytes. 7. Carefully cut longitudinal sections from one side of the female gameto- phyte until the embryo sporophyte comes into view, and sketch the section under dissecting microscope, showing the embryo in position. LXXIII. Conifers. General Study. Phylum, Strobilophyta. Class, Conifere. Order, Pinales. | The conifers called for below are cultivated quite extensively, and material for study can usually be obtained without difficulty. (a) Various Conifers. . Collect branches of the following: Pinacee—Norway spruce (Picea abies (L.), Canadian hemlock (Tstiga canadénis (L.), European larch (Larix larix (L.). Juniperacezee —arborvite (Thuja occidentalis L.) ~ 1. Sketch a short branch of the Norway spruce and note a slight tendency to bilateral symmetry, and how the leaves are bent from the under side to obtain a proper light relation. 2. Sketch a branch of the Canadian hemlock with carpellate cone at the end. Note bilateral arrangement and the light relation of the leaves, especially the small ones on the upper side. 3. Sketch the larch branch showing the large dwarf branches. Compare with Ginkgo. Note that the foliage leaves are deciduous annually, and that the dwarf branches may develop into ordinary branches. Are the dwarf branches deciduous (self-pruned) ? 4. Sketch a small branch of the arborvite. Note the flattened condition of . the stem and the leaves. Note also that numerous branches of various sizes are self-pruned. (b) Pinus. Family, Pinacez. Collect large branches of white pine (Pinus strobus L.) pitch pine (P. rigida ll.) Austrian pine (P. laricio Poir.), and Scotch pine (P. silvéstris L.) Also lect the dwarf branches with needle-leaves which have been self-pruned. 1. Study and sketch a branch of the Austrian pine, showing scale leaves, dwarf branches, and foliage leaves (needles). How old is the branch studied? What two ways of telling the age? Are the foliage leaves deciduous? How old are the dwarf branches before they are self-pruned? Where do the ordinary branches originate, and when? M i col LABORAPRORY QULEINES FOR: GENERAL BOTANY. 67 2. Draw a dwarf branch, with scale leaves and foliage leaves, of the white pine, pitch pine, Austrian pine, and Scotch pine. Note the pecularities of each dwarf branch. Compare with Larix and Ginkgo. 3. Under low power, without cover-glass, draw part of the foliage leaf of the Austrian pine, showing the stomata. How are they arranged? Draw a scale leaf from the ordinary branch and one from the dwarf branch. Note use difference between the foliage leaves and the scale leaves. 4. Cut cross sections of a foliage leaf, mount and study under low power. Draw and note the following tissues: epidermis with sections of the stomata, heavy-walled hypodermal tissue, green mesophyll with a number of resin-ducts, a limiting layer of large clear cells, and the light-colored central region with two vascular bundles. (c) Structure of White Pine Stem. Preserve pieces of branches, one to six years old, in alcohol, and also obtain large, polished cross sections (about two inches thick) of a tree-trunk with bark. 1. With a strong, sharp razor, cut cross, tangential and radial sections of stems in alcohol, mount, and stain with iodin; or study prepared slides. 2. Draw part of a cross section under low power, showing epidermis, cortex, with resin passages, phloem, cambium, xylem in a number of annual rings with medullary rays and resin passages, and central pith. 3. Radial section. Draw under low power, showing cortex, cambium, xylem (composed of the tracheids), and pith. Note the medullary rays passing from the pith to the phloem. 4. Tangential section. Draw under low power, showing part of the xylem with tracheids and cross sections of the medullary rays. 5. Under high power draw part of a tracheid from radial section, showing the peculiar bordered pits. 6. Sketch part of a polished section of an old pine stem, showing bark, cambium, sap wood, heart wood, and pith. Notice the medullary rays. Notice also that each annual ring of wood is double— early wood and late wood. On which side is the early wood? Describe the growth of a pine tree in height and thickness. (d) Sporophylls of Pinus laricio. Use fresh or alcoholic material. 1. Draw a staminate (microsporangiate) cone under dissecting microscope. Describe the arrangement of the stamens (microsporophylls). 2. Draw a stamen under low power, showing the outer (under) side with two microsporangia (pollen-sacs). How different from the microsporophyll of Selaginella in structure and function? 3. Draw a young carpellate (megasporangiate) cone under dissecting micro- scope. Describe. Note that the parts are smaller at the lower end. 4. Draw a carpel (megasporophyll) from the lower side under low power, showing the bract (true leaf blade of the carpel) and the large ovuliferous scale. This may be an outward growth of the chalazal region of the ovules. Draw the carpel from the inner (upper) side, showing the two ovules (megasporangia) and the ovuliferous scale. Compare the carpel with the megasporophyll of Selaginella. 5. Draw a mature carpellate cone. Note the spiral arrangement and that the carpels at the base are undeveloped and contain no seed. This is an example of rudimentary organs. If a rudimentary organ was formerly more _ highly developed and functional it is called a vestigial organ or a vestige. 68 LABORATORY OUTLINES FOR GENERAL BOTANY. 6. Notr.— The staminate and carpellate cones of the pine represent primitive flowers. Are these flowers monosporangiate (one kind of spores in the flower) or bisporangiate (both kinds of spores in the same flower)? Compare with the cone in Selaginella. Is the pine tree (sporophyte) monecious or diecious? (e) Carpellate Cone of Larix larix. Collect carpellate cones of the usual type and some which have the tip continued as a leafy branch. Preserve in alcohol. 1. Sketch a normal cone in which terminal growth has been completely checked. ' 2. Sketch a cone on which a leafy branch has developed at the outer end. Note the gradual transition from carpels to. ordinary foliage leaves. Sketch a number under dissecting microscope, showing this transition. This continued growth or prolongation of the floral axis of the larch cone is a good example of reversion to a more primitive condition or atavism. Compare with the ordinary ferns, Lycopodium lucidulum, and Cycas. 3. Observe fresh or dried, young cones and note the presence of a special | color. How do you account for the color in this cone? ({) Gametophytes and seed. The gametophytes of Pinus laricio may be studied from staminate and car- pellate cones preserved in alcohol. The seeds may be kept in a dry condition. 1. Draw a male gametophyte (pollen-grain) under high power. Note the two wings. These represent an adaptation for anemophilous pollination. 2. Remove the female gametophyte from a young seed (collected at the time of fertilization, about July 1), and draw under dissecting microscope. Note the difference in size between the male and female gametophytes. Compare the two gametophytes with those of Marsilea and Selaginella. 3. Draw a mature seed. Remove the testa and the inner seed coat. What does the inner coat represent? Draw the female gametophyte. Carefully cut out the embryo sporophyte, which is now in a dormant condition. Sketch under dis- secting microscope, showing the radicle, suspensor and cotyledons. Pick off the cotyledons from one side and sketch the plumule. How many cotyledons? Instead of P. laricio, the seeds of Pinus edulis Engelm, the nut pine of commerce may be used to advantage. 4. If prepared slides are at hand draw a section of a stamen, showing the one-called microspores. — 5. Draw a section of a male gametophyte (or study mature pollen-grains preserved in alcohol) showing the large tube cell and nucleus, the generative cell and the two disorganized vegetative cells lying like two thin plates against the wall of the grain back of the generative cell. 6. Draw a section of a young ovule, showing the functional megaspore. 7. Draw a pollen-grain which has formed a short pollen-tube growing down into the nucellus (tip of the megasporangium). Note the tube nucleus in the tube and in the body of the grain the spermatogenous cell, the stalk cell and the remains of the two evanescent vegetative cells. The spermatogenous cell divides later into two sperm cells which do not have flagella or cilia. From the same section draw the spherical embryonic female gametophyte. &. Draw a female gametophyte showing archegonia (ovaries) with ospheres. 9. Draw an archegonium (ovary) in which the nucleus of the oosphere has divided into four nuclei. LABORATORY OUTLINES FOR *GENERAL BOTANY. 69 10. Draw the upper part of a female gametophyte, showing remains of arche- gonia with an elongated cavity below them in which appear a number of embryos in various stages of development. Only one of these embryos survives, probably the one which has a slight advantage in size, vigor, and food supply. Note the struggle for existence which must go on among these embryos. 11. Sketch a mature seed, showing the wing. Let a winged seed drop to the floor from a height of six or seven feet and note how it falls. Describe the . adaptation this seed has for dissemination. Note also the readiness with which the seed is separated from the wing. Of what use is this adaptation? Pie 4 irs CyveLre or Pine. (g) Seedlings and Primitive Leaf Arrangement. Plant seeds of Pinus and Thuja accidentdlis and use fresh plantlets or pre- serve in alcohol. Also obtain branches of the common juniper (Juniperus com- munis L. Family, Juniperacee) and cultivated varieties of Thuja known as retinispora forms. In these retinispora or juvenile forms, branches often change suddenly from the form with spreading leaves to the flattened condition, and the flattened branches again revert to the form with spreading leaves. 1. Sketch a pine seedling which has sprouted, showing the seed coat still covering the cotyledons. Sketch a seedling with cotyledons expanded. Describe the important changes which take place in the embryo during the process of sprouting. 2. Sketch a branch of Juniperus communis and note that all of the leaves are of the spreading type. 3. Study and sketch the seedings of Thuja occidentalis and note that at first the leaves are of the spreading type much like those of Juniperus, and that later the branches have the flattened form characteristic of the adult plant. Apply the recapitulation theory as given in connection with the moss protonema. From 70 LABORATORY) OUTLINES FOR (GENE KAI sD Omi ee this it would appear that the ancestors of Thuja had the leaves arranged like those of the common juniper. 4. Study and draw a small branch of Thuja (retinispora form), in which the upper part of a flattened branch has changed back to the juvenile form. In such cases there is a second reversion. In other words, the branch takes on first one form and then another successively. 5. Make a diagram showing the life cycle of a pine. See Fig. 14. 6. ‘Nore on the development of the carpellate pine cone. The young carpellate cones of Pinus laricio begin to develop in the bud during the summer or fall, and in the following spring the carpels have young ovules with a distinct integument. Later (about the middle of May) the ovules are pollinated and the megaspore is developed. In the following autumn (Octo- ber) the megaspore has germinated and the female gametophyte is developing as a hollow spherical body composed of free, naked cells. It passes the winter in this condition. In June of the following year the archegonia with eggs are ready for fertilization and the pollen-tubes have grown down ‘thru the nucellus. About the last week in June or the first in July fertilization occurs, and the embryo is matured and in the resting condition in the following autumn. The seed is usually shed late in the winter or in the early spring of the year following. The whole history thus covers nearly three full years. LXXIV. Taxus canadénsis Marsh. American Yew. Phylum, Strobilophyta. Class, Conifere. Order, Taxales. Family, Taxacez. The yew is a low shrub growing on moist banks and hills, especially in the shade of large conifers. It is common northward. Herbarium and alcoholic material may be used if fresh branches are not available. 1. Sketch a branch, showing arrangement of leaves. Describe. 2. Under dissecting microscope draw a staminate cone. How are the stamens arranged? 3. Draw a single stamen under low power and note the peltate form. How many microsporangia? Compare the shape of this stamen with the sporophyll of Equisetum. 4. Under dissecting microscope draw a small fertile branch with a young ovule at the tip. 5. Cut longitudinal sections of the branch with ovule, mount, and draw under low power, showing the megasporangium in the center surrounded by the long inner integument and a short outer undeveloped aril, with scale-leaves on the stem below. 6. Draw a ripe seed with the thick, fleshy, red aril. Compare the aril with the ovuliferous scale of Pinus. LXXV. Higher Gymnosperms. Phylum, Strobilophyta. Class, Gnetee. Study herbarium specimens. 1. Make a sketch of a small plant of Ephédra Sp. (Order, Ephedrales. Family, Ephedracee). Note the slender green stems and the dry scale-leaves. In what ways is this plant adapted to a xerophytic environment? 2. Make a sketch of a branch of Gnétum gnéemon L. (Order, Gnetales. Family, Gnetcee).. Note the large broad leaves. This is a tropical tree cul— tivated in India and surrounding regions. WAT OKADROK VS OULLEINES (FOR GENERAL BOLAN V., 71 SUB-KINGDOM, ANGIOSPERMZ. A NUMBER OF FORMS TO REPRESENT THE GENERAL EVOLUTION OF THE FLOWER IN MONOCOTYLS AND DICOTYLS. Along with this series of outlines on the Anthophyta, the student should be given work in identification with a key and several periods can profitably be spent — in analyzing and making diagrams of various spring flowers. LXXVI. Magnolia sp. Magnolia. Phylum, Anthophyta. Class, Dicotyle. Order, Ranales. Family, Magnoliacee. | The magnolias are among the most primitive of the Anthophytes. Any of the native or cultivated species will have suitable flowers in early spring. They may be used fresh or preserved in alcohol. Sporophyte. 1. Sketch the entire flower; describe size, color, etc. Note the character of the stem. Compare the flower with the cones of Lycopodium, Selaginella, and Pinus. 2. ketch a sepal, a petal, a stamen and a carpel; describe each organ. The stamen is a microsporophyll and the carpel a megasporophyll. 3. How many sepals in the calyx? How many petals in the corolla? How many stamens in the andrecium (stamen set)? How many carpels in the gyne- cium (carpel set)? How many cycles in the perianth? Note especially that the stamens and carpels are arranged spirally. Compare several flowers as to the constancy or variability in number of parts. Make a diagram of the flower. See Fig. l6a. LXXVII. (a) Sagittaria latifolia Willd. Arrow-head. Phylum, Anthophyta. Class, Monoctyle. Order, Alismales. Family, Alis- macex. The broad-leaf arrow-head grows in moist ground on the margin of ponds, creeks and canals and blooms in summer. If fresh material is not available, good herbarium specimens may be used. Flowers and other parts may also be preserved in alcohol. Sporophyte. 1. Sketch and describe the entire plant, noting the character of the leaves, stem, roots, and inflorescence. 2. Sketch the staminate flower, showing sepals, petals and stamens. How many parts in each set? Find the vestigial carpels. Draw one under dissecting microscope. 3. oketch the carpellate flower and describe the parts present. What parts of the two flowers are cyclic and what parts spiral in arrangement? Is this sporophyte monecious or diecious? 4. Under dissecting microscope draw a sepal, a petal, a stamen (microsporo- phyll) and a carpel (megasporophyll). Compare the normal carpel with a ves- tigial carpel. How did this plant attain the monecious condition? 5. Cut cross sections of a young stamen, mount, and draw under low power. How many microsporangia? Note that the stamen is made up of anther and filament. 6. Cut off one side of a carpel so as to expose the ovule (megasporangium). Draw under low power, showing the stigma, short style, and ovulary. Note 72 LABORATORY OUTLINES FOR GENERAL BOTA: that the stigma is a new organ not present in any of the Gymnosperms. Why is the stigma necessary to this carpel? . (b) Ranunculus abortivus L. Crowfoot. Phylum, Anthophyta. Class, Dicotyle. Order, Ranales. Family, Ranuncu- lacez. This plant is common in April and May along brooks, on hillsides, in meadows, and along roads. Sporophyte. 1. Sketch the entire plant, showing the various organs. 2. Sketch the flower and describe the condition of the four sets of floral organs. Note that the flower is bisporangiate. Compare with the cone of Sela- ginella. 3. Draw a sepal, a petal, a stamen, and a carpel under dissecting microscope. LXXVIII. Alisma subcordatum Raf. Water Plantain. Class Monocotyle. Order, Alismales. Family Alismacez. The water plantain is common in wet and muddy places, on the margin of ponds and creeks and blooms in summer. Herbarium specimens and preserved material may be used. Sporophyte. 1. Sketch a leat and a part of the inflorescence. 2. Sketch a flower showing the four sets of floral organs — calyx, corolla, andrecium and gynecium. How many sepals, petals, stamens and carpels? Are the parts spiral or cyclic? Free or united? Is the flower monosporangiate or bisporangiate? Note that the flower is hypogynous. What advance does this flower show over that of Sagittaria or Ranunculus? 3. Make a diagram of the flower. See Fig. 16b. 4. Cut cross sections of the stamens or, use prepared slides and draw under low power. How many microsporangia (pollen-sacs)? Cut open the ovulary and dissect out the ovule (megasporangium). Draw. 5. From prepared slides draw a microsporocyte and a microspore, showing the nucleus, cytoplasm and wall. 6. From prepared slide draw a young ovule, showing the funiculus, the integ- uments, the megasporangium proper (nucellus), and the single megaspore. Note the absence of a wall around the megaspore. Why not present? Gametophytes. 7. From prepared slide draw a male gametophyte (pollen-grain), showing the tube nucleus and the two elongated sperm cells. 8. From prepared slide draw an eight-celled female gametophyte (embryo- sac), showing the three anipodal cells, the two polar nuclei, the oosphere, and the two synergids. The oosphere and the two synergids are called the egg- apparats (ovary). 9. From prepared slide draw a mature, seven-celled female gametophyte, showing the fertilization of the egg and the conjugation of the polar cells to form the definitive cell. Look for the conjugation of the second sperm with the polars (triple fusion). 10. From a prepared slide draw an embryo-sac with endosperm cells, which have come from the division of the definitive cell, and with young embryo, con- sisting of the embryo proper, the suspensor cells and the large, vesicular, basal, LABORATORY QUYTLINES FOR GENERAL BOTANY. 73 suspensor cell. Note that the conjugation of the polar cells and the subsequent development of the endosperm are entirely new phenomena, nothing similar being known in plants below the anthophyta. In many other anthophyta the second sperm cell from the pollen-tube comes down and unites with the polar cells pro- ducing a triple fusion, as in Alisma. 11. Carefully remove a mature embryo from the seed and sketch under low power, showing the single cotyiedon, the lateral plumule and the radicle. Fic. 15.— Lire Cycle or ANGIOSPERM (ALISMA.) 12... Note that in ‘this plant the seed remains in the ovulary, i. e. the fruit is indehiscent. Make a diagram showing position of the carpel wall, the integu- ments of the ovule, the endosperm and the embryo. Note also that the ovule is first orthotropous, then anatropous and finally campyllotropous. 13. Sketch a young seedling. 14. Make a diagram in the notes showing the general life cycle of an angiosperm. See Fig. 15. LXXIX. Sédum acre L. Wall-pepper. Class, Dicotyle. Order, Saxifragales. Family, Crassulacee. Many of the sedums grow well in greenhouses and in window gardens. They usually bloom abundantly in the spring and the above or any other species will be found suitable. 1. Make a careful drawing of the flower and describe the character of the different parts. 2. Make drawings of the calyx, the corolla, the andrecium and the gynecium. 3. Answer the following questions correctly: Is the flower hypogynous, perigynous, or epigynous? Is it tetracyclic or pentacyclic? Are the circles or whorls trimerous, tetramerous or pentamerous? Z 74 LABORATORY OUTLINES, FORSCENERAL BOT7INie Are the organs of any whorl or set united or partly united? Is the flower isocarpic or anisocarpic? Is it actinomorphic, isobilateral, zygomorphic, or unsymmetrical? 4. Make two diagrams showing the true condition of the flower as learned above. See Fig. 16 c and d. LXXX. Trillium grandiflorum (Mx.). Large-flowered Trillium. Class, Monocotyle. Order, Liliales. Family, Liliacee. The large flowered Trillium grows in rich woods and blooms in April and May. 1. Make a sketch of the entire plant, showing the flower, leaves, and short tuberous rhizome with contractile roots below. How deep was the rhizome under ground? Describe how it descends into the earth. This plant is a geophilous, herbaceous perennial. What are the advantages of the geophilous habit? 2. Cut across section of the compound ovulary, mount, and draw under low power, showing the cavities with ovules. 3. Describe the condition of the flower according to the questions asked under Sedum acre. Make a diagram of the flower. See Fig. 16e. LXXXI. Cypripédium parviflorum Salisb. Yellow Lady’s-slipper. Class, Monocotyle. Order, Orchidales Family, Orchidacez. This lady’s-slipper grows in wet places and low woods, and blooms in May and June. Any other species will do as well. 1. Sketch part of a plant, showing the flower and part of the leafly stem. Fic. 16.— DIAGRAMS OF FLOWERS. 2.. Cut cross sections of the ovulary, mount and draw. How many carpels? Study the flower with the aid of the diagram, Fig. 16 f. HABORANORY OULEEINES FOR GENERAL. BOTANY, 75 3. Copy the diagram in the notes and write a general description of the flower, noting especially that it is organized on the same plan as the Trillium flower, that some of the parts have disappeared, that it is epigynous and zygo- morphic, that certain parts are united, and that it is highly specialized for insect pollination. 4. Why should this flower be placed higher than any of the monocotyls pre- viously studied? Make a comparison of the flower of Sagittaria, Alisma, Trillium and Cypripedium. LXXXII. Catalpa speciOésa Warder. Hardy Catalpa. Class, Dicotyle. Order, Scrophulariales. Family, Bignoniacez. The Catalpa is cultivated extensively and blooms abundantly in May and June. 1. Study the large compound panicle and draw a single flower. 2. Describe the flower carefully, noting the condition of each floral set and whether the flower is hypogynous or epigynous, whether actinomorphic or zygo- morphic. What adaptations for insect pollination? Note especially the rudi- mentary or vestigial stamens. Be careful to distinguish vestigial organs (vestiges) from incipient organs (incepts) and from nascent organs. 3. Cut cross sections of the ovulary,, mount, and draw. How many carpels in the gynecium? 4, Make two diagrams of the flower, showing transverse and longitudinal arrangement. LXXXIII. Cornus fémina Mill. Panicled Dogwood. Class, Dicotyle. Order, Umbellales. Family, Cornacez. This common shrub usually forms thickets, in forests and on hillsides. It blooms in June, producing an abundance of flowers. Any other species with panicled flowers will do. 1. Sketch the entire infloresence and note the arrangement of the numerous small flowers. 2. Under dissecting microscope draw a single flower. How many cycles? How many stamens, petals and sepals? Note the minute size of the calyx. 3. Cut cross section of the ovulary. Draw under low power. How many carpels? Note that the flower is epigynous. 4. Compare this inflorescence with that of the flowering dogwood, ‘Cynoxylon floridum (L.) noting especially the origin and nature of the white involucre. 5. Make a transverse and a longitudinal diagram of the flower. See Fig. 16 g and h. ; LXXXIV. Ageratum conyzoides L. Ageratum. Class, Dicotyle. Order, Compositales. Family, Helianthacee. Ageratums are annuals which bloom all summer and are much used for borders. The flowers may be had in the greenhouse at any time of the year. The plants will live and bloom for a long time. 1. Sketch one of the heads under dissecting microscope, showing the bracts of the involucre and the numerous small tubular flowers. 2. Under dissecting microscope draw a single flower. What is the condition of the pappus? What does the pappus represent? 3. Dissect the flower and draw the corolla and andrecium and the gynecium under dissecting microscope or low power. Describe the flower and its parts in detail. 76 LABORATORY OUTLINES FOR GENERAL BOTANY. LXXXV. Chrysanthemum leucanthemum L. Ox-eye Daisy. Order, Compositales. Family, Helianthacez. This plant grows in fields and meadows and blooms in May and June. - - 1. Draw one of the heads, showing the bracts of the involucre, the lingulate or ray flowers and the tubular or disk flowers. 2. Under dissecting microscope draw a ray flower and a disk flower. Describe each. What is the condition of the calyx? Why should the outer flowers develop as ray flowers rather than the inner ones? Note that the ray flower is zygomor- phic. LXXXVI. Leéntodon taraxacum L. Dandelion. Order, Compositales. Family, Cichoriacee. The dandelion blooms from early spring to late autumn, so plants may usually be obtained without difficulty. 1. Sketch an entrie plant, showing root, short stem, rosette of leaves, and slender stems bearing heads of flowers. Note that the dandelion is geophilous. Why does it not grow up out of the ground? How do you account for the rosette habit? Remember that the lowest seed plants were trees. This is one of several culmination types. 2. Make a sketch of a single head. Note that all the flowers are ligulate. Also note that the embryonic corollas are tubular, showing five teeth on the limb, and only become strap-shaped when they expand. Make a series of sketches show- ing this. How does this indicate that Leontodon is a higher type of development or specialization than Chrysanthemum leucanthemum? . 3. Under dissecting microscope draw a single flower. Describe the pappus, corolla, andrecium, and gynecium. 4. Draw some of the ripe furit. ‘Note adaption for. suspension in the air. Of what special advantage is this parachute arrangement? Note the action of the involucre while the fruit is ripening. This extraordinary parachute is an ex- ample of overadaptation. ®. How many seeds are in each dandelion fruit? i. e., how many for each flower? How many seeds are produced on an average-sized head? About how many heads of flowers are matured from a fair-sized dandelion plant in one sea- son? 6. Suppose that you had but one mature dandelion plant and that it produced seed normally for ten years and that each seed developed into a mature plant and began to reproduce at the average rate the second year, (i. e., when two years old) how many offspring would there be at the end of ten years? 7. The total land surface of the earth is about fifty-three millions of square miles, how many dandelion plants would there be for each square mile of land surface at the end of ten years? 8. Nore.— The above problems will indicate to some extent the great possi- bilities of reproduction present in many plants. It will be remembered that each seed contains a little, dormant embryo; therefore, every seed that perishes means the destruction of a young plant. It is evident that a very large per cent, of young plants must perish each year, and that those which survive for any length of time must usually undergo a severe struggle for existence. In this struggle for life and place the fittest usually survive; i. e., those which are able to grow more vigorously and thus crowd out thei- weaker neighbors and those which are best adapted to their environment. LABORATORY OUTLINES FOR GENERAL BOTANY. 77 OTHER TYPES OF ANGIOSPERMS. LXXXVII. Triticum aestivum L. Wheat. (T. vulgare). Phylum, Anthophyta. Class, Monocotyle. Order, Graminales. Family, Gram- inacez. a Material should be collected during and after the flowering period in May and June, and dried or preserved in 70 per cent. alcohol. 1. Study and sketch the spike (head) and describe. Note the character of the stem. What mechanical advantage in the disposition of the tissues of the stem? The axis of the spike is called the rachis. 2. Draw a single spikelet, noting the two empty glumes at the base, and several flowers. How many flowers? What is the condition of the uppermost flowers of the spikelet? The axis of the spikelet is called the rachilla. 3. Draw the two empty glumes noting their peculiarities. 4. Dissect a single flower. It is inclosed in two glumes, the flowering glumes; the one with the awn is the lemma, and the inner one is the palet. Draw both and describe. 5. Just inside of the lemma at the base of the gynecium are minute scales called lodicules. How many? Draw one under low power. What might they represent? 6. There are three stamens in the andrecium, and the gynecium develops into a single grain. Make a diagram showing the positions of the empty glumes, the lemma and palet, the lodicules, the stamens, the ovulary, and the axes of the spike and spikelet. LXXXVIII. Ftchsia sp. Fuchsia. Class, Dicotyle. Order, Myrtales. Family, Onagracee. Fuschias are commonly cultivated in greenhouses and as house plants. The flowers should be studied fresh and any of the common greenhouse species will do. 1. Study and draw the flower. 2. Draw the gynecium with style and stigmas. Cut cross sections of the ovulary, noting the number of cavities and ovules. 3. Draw the perianth tube or hypanthium split open, showing the calyx, corolla, and andrecium. How are these parts grown together? Note the color of the calyx and the corolla. 4. Why does the epigynous condition and the peculiar development of the hypanthium indicate a high type of floral development? Describe the entire flower. 0. Make transverse and longitudinal diagrams of the flower, showing the relationships of the parts. LXXXIX. Pépulus deltoides Marsh. Cottonwood. Phylum, Anthophyta. Class, Dicotyle. Order, Salicales. Family, Salicacee. This is a large tree of rapid growth, common on flood-plains of rivers, and is much planted for ornament. It blooms in April. 1. Collect staminate and carpellate catkins or aments, noting that the trees are diecious—some are staminate trees and some carpellate trees. Note the differ- ence in color between the two kinds of flowers. How do you explain the differ- - ence? Draw the two catkins. 78 LABORATORY OUTLINES .FOR GENERAL BO TANa. 2. Draw a single staminate flower and a carpellate flower and describe. Any perianth? Why are the stamens red and not yellow as is commonly the case? Why is it incorrect to say male tree instead of staminate tree and female tree instead of carpellate tree? Or why should you not say male flower and female flower? 3. When the capsules ripen study the seed. How is it distributed? How effective is this method? Why should only staminate trees be planted in a city? How would you make sure that you had staminate trees to plant? Could you plant cuttings? This tree endures city conditions quite well. 4. iNote the different kinds of scars on the tree: leaf scars, stipular scars bundle-scars, self-pruning scars, lenticels. Note also that the pith is 5-angled. Is there anything on the outside of th twig corresponding to this? 9. Note that the leaves have a strong tendency to take a vertical position especially on twigs that grow erect. Draw and describe the flattened condition of the petiole by which this is accomplished. Why are the two sides of the leaf nearly alike? What advantage in the vertical position? ‘Why do you hear a musical rustle of the leaves when the wind is blowing? Note the two glands at the base of the blade. Of what use are the glands? 6. Study and draw self-pruned branches.” Fresh material can be obtained in summer and autumn and preserved in alcohol but the dry twigs will do fairly well. Notice that the winter buds are in perfect condition. Draw the base showing the surface of the scar. Draw a self-pruning scar that has healed over. How is the cleavage plane produced in the basal joint? Why are the branches pruned off? How old are the branches when self-pruned? 7. Note. This outlin may be used as a special exercise to be worked up during the term. A special paper may be written on the subject including the above and many other interesting points connected with the life of this tree. XC. Polemonium réptans L. Greek Valerian. Class, Dicotyle. Order, Polemoniales. Family, Polemoniacee. This is a perennial herb growing in woods. It blooms in April and May. Fresh material must be used. 1. Describe the entire plant and sketch a branch showing the leaves and flowers. If young plants are available, note the circinate vernation. 2. Study a single flower. Note the blue color. Is blue a common color of flowers? Note the character of the calyx, the corolla, the andrecium, and the gynecium, and draw and describe each set. 3. Cut cross sections of the ovulary and draw. How many cavities? How many ovules in each cavity? 4. If ripe capsules are present wet the seeds on the slide and examine care- fully under low power. What peculiarity becomes evident after a few moments? Crush or break up the seed with a scalpel and observe further. 5. Make a diagram of the flower. SPECIAL EXERCISES ON ANGIOSPERMS. XCI. Comparison of Carpels. 1. If not previously studied, draw a carpel of Cicas revoluta L. 2. Draw a carpel of the Kentucky coffee-bean (Gymndécladus didica (L.).) 8. ketch a nearly mature fruit of the Velvetleaf (Abutilon abutilon (L.).) Separate the ovularies and draw a single carpel. PABORATORY OUTEINES POR GENERAL BOTANY: 79 4. Carefully separate the ovularies of the carpels of an orange (Citrus auran- tium LL.) so they will lie side by side in a row. Draw. Note that some of the divisions are smaller than the normal. There is a struggle for existence among the members of the gynecium so that some are not fully developd. 5. Make a comparison of the four fruits studied above. XCII. Dicotyl Seed. Study the fruit and seed of the Olive (Olea europaéa L.). Use either fresh or pickled olives. The samara of Fraxinus is also satisfactory. 1. Sketch the entire drupe. Note that the pericarp consists of a fleshy exocarp and a thick, stony endocarp. 2. Break the endocarp and remove the fleshy seed within. Remove the mem- branous seed coats and sketch the fleshy kernel inside. This is the endosperm. 3. Remove the embryo and draw, showing the short radicle, the two cotyle- dons, and the plumule. Compare the kernel of Olea with that of the Pine seed and note the differences. XCIII. Section of Leaf. (a) 1. Cut cross sections of the lamina-of a sunflower (Helianthus dnnuus Ee )pvlcat- fresh -or presenved in alcohol, mount, and study under low and high power. : 2. Draw and describe, showing the following tissues: upper epidermis with thick cuticle and multicellular hairs, palisade parenchyma, spongy parenchyma with large intercellular spaces, sections of vascular bundles, and lower epidermis with stomata and multicellular hairs. 3. How do. you account for the palisade arrangement of the cells in the upper part of the leaf ? (b) Cut sections of Beech Leaves or use prepared slides. Draw and com- pare the tissues with those in the sunflower leaf. XCIV. Leaf Variation. 1. Obtain a series of fresh or pressed leaves of the red mulberry (Morus rubra L.) or of the giant ragweed (Ambrosia trifida L.) and make outline sketches of ten different forms. XCV. Section of Winter Bud. 1. From alcoholic material cut longitundinal sections of common lilac buds (Syringa vulgaris L.) Mount and sketch under low power. Note the flat apex with outer dermatogen and hypodermal meristematic tissue; and a little farther down the epidermis, cortex (periblem), procambium (formative tissue of vascular bundles), and the central pith. 2. Note the origin of the leaves, beginning at the apex, and also the origin of the lateral buds in the axils of the leaves. Make a sketch showing the entire upper part of the bud, with all the structures mentioned above. XCVI. Monocotyl Stem. 1. Cut cross sections of young corn stems preserved in alcohol, stain and mount; or use prepared slides. Sketch the entire section under dissecting micro- scope, showing epidermis, band of sclerenchyma, large pith or ground tissue, and the scattered vascular bundles. [ 80 LABORATORY OUTLINES FOR GENERAL BOTANY. 2. Under high power draw one of the bundles. Note the large vessels situated in the xylem arranged like a letter V, the cavity in the tissue at the apex of the V, the bundle of phloem between and beyond the two large vessels, and the sheath of sclerenchyma about the bundle. XCVII. Herbaceous Dicotyl Stem. (a) Sunflower Stem. 1. Cut cross sections of a young sunflower stem preserved in alcohol, mount and stain; or study prepared slides. 2. Sketch the entire section under dissecting microscope, showing cortex, circle of vascular bundles, and large central pith. This vascular system is an example of a siphonostele. Compare with Botrychium. 3. Under low power draw part of a section showing the epidermis with epidermal hairs, the layer of collenchyma immediately below this, the parenchyma with resin passages, the vascular bundles with cambium layer, the medullary rays, and the central pith. 4. Under high power draw a single vascular bundle, selecting one of the narrow, oval type. Represent in order the external bundle of sclerenchyma, the phloem, the cambium, and the xylem usually in a double layer. 5. Notice the mechanical principles involved in the structure of the stem and the vascular bundle. Compare with a T railroad rail. (b) Pumpkin Stem. 1. Cut cross and longitudinal sections of the stem of Cucurbita pepo L., stain and mount, or study prepared slides. Sketch the cross section under dissecting microscope. ‘Note the epidermis, the cortex, the vascular bundles, the pith, and the large central cavity. 2. Under high power study the longitudinal sections and draw sieve-tubes showing the sieve-plates in the phloem. 3. In the xylem find and draw one of the large reticulate wood vessels; also a spiral wood vessel with a single spiral thickening and one with two spirals; also draw an annular wood vessel in which the thickenings are in the form of rings. XCVIII. Dicotyl Woody Stem. 1. Cut cross sections of a very young twig of the beech (Fagus grandifoha, Ehrh.) and one a year old, preserved in alcohol, stain and mount, or use prepared slides. Under low power note the cortical layer, the circle of vascular bundles with medullary rays between, and the central pith. Draw. This is a siphonostele. Compare the younger section with the sunflower. Under high power draw a nar- row sector passing thru a vascular bundle, showing epidermis, cork (periderm), cork cambium (/phellogen), seoondary cortex (phelloderm), primary cortex, sclerenchyma, phloem, stelar cambium, xylem (wood), and pith. Draw a single complete bundle? showing the sclerenchyma, the phloem, the cambium and the xylem. 2. Cut cross sections of twigs of the White Ash (Fraxinus americana L.), preserved in alcohol, one very young, one a few months old, one a year old and one two years old. Study and draw the young sections noting the epidemis, cortex, vascular bundles and pith. Study the one and two year old sections and note the formation of the cork and the secondary development of phloem and xylem: Draw and describe. LABORATORY OUTLINES FOR GENERAL BOTANY. 81 3. Cut cross sections of linden twigs (Tila sp.) preserved in alcohol. Take one very young branch, one a year old, one two years old, and one three years old. Mount and stain; or use prepared slides. Study under low and under high power. Draw the year old section, noting the following structures: epidermis, cork layer, cortex, phloem and sclerenchyma layer (inner bark), stelar cambium, xylem (wood) with medullary rays, and pith. 4. Make a series of diagrams showing the primary structure and how the secondary structures are developed for the first three years. 5. Under high power draw a small area of the xylem showing a medullary ray, large empty pitted vessel, small empty wood fibers, narrow empty tracheids, and protoplasmic wood parenchyma. Draw also a small area of cells from the bark, showing a medullary ray, light-colored thin-walled sieve tubes, very thick- walled bast fibers with only a minute lumen, protoplasmic companion cells in and around the sieve tube tissue, and large protoplasmic bast parenchyma. 6. Cut cross sections of a very young and of a year old twig of cottonwood (Populus deltoides Marsh.); mount and stain. ‘Note the following structures: epidermis, periderm (cork layer) and the five corky ridges, phellogen (cork cam- bium), phelloderm (secondary cortex), primary cortex, sclerenchyma (fiber bun- dles), phloem with a narrow band of sclerenchyma, stelar cambium, xylem (wood), and the five angled central pith. 7. Make a sketch of a cross section of a polished tree trunk of black walnut (Juglans nigra L.), showing these structures: pith, heartwood (duramen), sap- wood (alburnum), annual rings with early and late wood, medullary rays, the stelar cambium and phloem (inner bark) separated by irregular strips or areas or corky tissue from the outer bark. The outer bark has been developed from the cortex and phloem and modified by successive layers of cork cambiums (phellogen). These irregular strips of corky tissue can easily be seen in the -outer bark with the naked eye. 8. Compare with the walnut a polished section of the trunk of a bur oak (Quércus macrocarpa Mx.) In this the medullary rays are much more prominent. 9. Study cross section of the trunk of Catdalpa speciosa Warder. Measure ‘the width of the annual rings and draw a curve showing the growth in diameter for the entire life of the tree trunk from which the section was taken. MCIXe The: Root: (a) Section of Buckeye Root. 1. Cut cross sections of one of the larger fleshy rootlets of Aesculus sp. (preserved in alcohol). Mount and draw under low power, representing the fol- ‘lowing structures: the four or more primary xylem bundles, four or more primary ‘phloem bundles alternating with the xylem, the beginning of the stelar cambium passing between the xylem and phloem, the endodermis or bundle sheath, and the broad cortex, with a superficial layer of cells known as the piliferous layer. 2. Cut cross sections of a somewhat older root which has turned brown, mount, and sketch the entire section under low power. Represent the following structures: the central strand of xylem composed of wood vessels and smaller cells, the stelar cambium, the band of phloem consisting of several kinds of cells, the remains of the endodermis, and the cortex and piliferous layer turned brown. s 82 LABORATORY OUTLINES FOR, GENERAL BOGAN. (b) Embryonic Root Tip. 1. Carefully remove the hard parts around the base of the embryo in a grain of corn (Zéa mays L.) and with a razor cut longitudinal sections of the radicle of the dormant embryo. The corn may be soaked in water for a while before cuting the sections, tho this is not necessary. 2. Mount the central sections in water and sketch under low power.. Note the following embryonic tissues: the outer scutellum, the root-sheath of a dark appearance inside of the scutellum, and the root tip inside of the root-sheath. The root tip is made up of the root-cap (of a light color), the dermatogen (a layer of large cells inside of the cap), the dark layer of periblem, the central light plerome, . and the growing point at the tip of the plerome. Below the dermatogen, at the apex of the root is the formative tissue of the root-cap, known as the calyp- trogen. It will probably not be distinct enough in these sections to trace out, but its position should be noted. (c) Root Hairs. 1. Sprout grains of corn on moist blotting paper in a box or under a bell- jar; after a few days the roots will be covered with root hairs. Sketch under dissecting microscope. ‘ 2. With a scalpel cut off some of the epidermis containing root hairs, mount and examine under high power. Draw and describe. 3. Under low power examine roots of young seedlings planted in soil and note the relation of the root hairs to the soil particles. C. Lenticels. 1. Examine and sketch the bark of a green and of a year-old elder stem Sambucus canadensis L. showing the surface covered with lenticels. 2. Cut cross sections of the bark, mount, and examine under low power. Sketch one of the lenticels. How and where do they originate? CI. Starch, Cellulose, Lignin, tannin, etc. 1. Cut a potato and scrape off some of the cells. Mount in water and study under high power. Draw some of the large starch grains present, showing the hilum and the stratified structure. 2. Place a drop of iodin solution beside the cover-glass and watch its effect on the starch. What is the color reaction? 3. Mount some wheat flower in water and treat with iodin. Note the blue colored starch and the yellow colored proteid material. 4. Mount a hair of common cotton (Goss)pium herbaceum L.) It is made up of nearly pure cellulose except the small central cavity in which is a small amount of dry protoplasm. Draw. Treat with Schulze’s solution (Chlor-zinc- iodin) and after a while note the color reaction. Care must be taken so as not to get any of this solution on the microscope as it is strongly acid. 5. Treat cross sections of a sunflower stem (from alcoholic material) with Schulze’s solution, examine, and note the cellulose reaction in the walls of the cortical and pith cells. 6. Treat a section of a sunflower stem with phloroglucin, mount and study color reaction in the xylem bundle. Care must be taken in its use-as it contains an acid. 7. Cut cross sections of a young twig of linden preserved in alcohol, treat with phloroglucin, mount, and note color reaction in the wood. LABOKRAROKY (OUREINES FOR (GEN BPRAL-BOTANY. 83 8. Cut cross and tangential sections of peach pits taken just when they are beginning to harden. Mount and study the cells of this stony tissue. Draw a number of the cells and describe. 9. Tannin, which is the cause of astringency of many fruits, may be readily demonstrated by Vinson’s method. Pour some sweet spirits of nitre, or a 20 per cent. alcoholic solution of nitrous ether, into a glass stoppered jar containing broken pieces of pottery or glass to prevent the fruit to be treated from falling into the liquid, and then drop in some fruit rich in tannin like green dates or persimmons. Stopper tightly and let remain from 12-24 hours. At the end of that period the tannin bearing cells will be of a dark brown color and will show distinctly when the fruit is cut open. Mount some of the dark cells in water and examine under low and high power. Note the large tannin masses. Crush the masses by pressing on the cover-glass and note how they stretch and finally break. A stained fruit may be preserved in 95 per cent. alcohol. Date fruits are ideal for this experiment and may be obtained in autumn from Arizona. 10. Aleurone grains. Cut sections of the endosperm of a seed of Ricinus communis L., castor-oil plant, after soaking in water. Mount in alcohol or dilute glycerin and draw several cells showing the oval aleurone grains, Each grain usually has a crystalloid and a globoid in its interior. CII. Crystals. The material for sectioning may be preserved in alcohol. 1. Cut cross sections of the rhizome of the large blue-flag (/ris versicolor L.) mount, and under high power draw the simple crystals present. 2. Cut sections of a year old twig of the wahoo (Eudnymus atropurpureus Jacq.), mount, and draw the large compound sphere-crystals in the pith and cortex. 3. Cut sections of the rhizome of the lily-of-the-valley (Convallaria majalis .L.), mount, and draw the bundles of the needle-shaped crystals, raphides. 4. Cut cross sections of the leaves of the India-rubber fig. (Ficus eldstica Roxb.) mount and draw the large cystoliths which are amorphous masses of mineral substance suspended from a pedicel. The mineral substance of the cysto- lith is mainly calcium carbonate. , CIII. Lipochrome. 1. Cut thin sections of the rind of an orange, mount in water, and examine under high power. Draw a cell showing the chromoplasts. 2. Cut sections of the root of the common cultivated carrot (Daucus carota L.). Mount and draw a cell showing the color bodies. 3. Mount pieces of the yellow corolla of the squaw weed (Senécio aureus L.) or any other yellow flower, examine under high power and draw a cell with chromoplasts. Describe the cause of the yellow color in these tissues. CIV. Anthocyan. 1. Cut sections of the root of the red garden beet (Béta vulgaris L.), mount, and examine under high power. Note that the red coloring matter is in the cell sap. 2. Cut sections of any leaf with red color as the red leaved coleus (Coleus blumei Benth.), mount, and study the color under high power. 3. Cut off some of the epidermis of a red apple (Malus malus (L.),) mount, and study the cause of the color. 2 84 LABORATORY. OUTEINES POR GENERAL BO RAN YT. 4. Mount part of a petal of a red greenhouse Pelargonium. Study the red coloring matter in the cells. 5. Mount part of a petal of a blue flower like Sdlvia pitcheri Torr. or Viola odorata L.) and study the nature of the color. CV. Solution of Anthocyan. 1. Take a quantity of the corollas of Mauwrdndia barclaidna Lindl. (a com- mon greenhouse vine) or flowers of Tvradescantia virginica L., place them in a dish and after crushing them cover with a quantity of 95 per cent. alcohol. After a few days or so pour off the alcohol into a bottle and preserve. 2. Take a test-tube about one-third full of the alcohol and add a few drops of aqua ammonia. ‘Note color reaction. Neutralize with hydrochloric acid until the liquid is again clear. Continue to add acid drop by drop. What is the color? 3. Place some red pelargonium (greenhouse geranium) flowers directly in ammonia water. Note that they change to blue. Transfer to acid alcohol and note that they change back to red. 4. How do you account for the change of color in many flowers during the period of blooming and for the many varieties of color produced by cultivation as. in the common morning-glory (Jpomoéa purpurea (L.)? CVI. Temperature Test with Anthocyan. 1. Take two good thermometers which register alike, wrap the bulb of one in a red begonia leaf and the other in a green begonia leaf, put each in a tumbler and place for some time in direct sunlight. Note the difference in temperature. Place the tumbler’s with thermometers in diffuse light and note the temperature again. Place them in a dark box and after a while read the temperature. Make a second test in the sunlight. 2. Describe one of the uses of anthocyan in roots, stems, leaves, flowers and fruits. CVII. Chlorophyll Solution. 1. Take a quantity of green leaves, such as the blue grass or greenhouse pelargonium; place them in a porcelain mortar or other suitable dish; cover with 95 per cent. alcohol; and crush the leaves thoroly. After the alcohol is colored a dark green filter into a bottle and keep in a dark place. 2. Take a small quantity in a test-tube and examine by looking thru it toward the window. Note the deep green color produced by the transmitted light. Examine it by reflected light, by standing between the window and the tube, and observe that the color of the solution appears a deep dull red, some- thing like blood. . 3. Take a small quantity of the solution in two test-tubes, and place one in the sunlight and the other in a dark box. How long before the one in the sun- light fades out? Compare it with the one in darkness. Thus it will be seen that sunlight when too intense will rapidly change the character of chlorophyll, altho it is generally absolutely necessary for its development. CV Iii; tilatex: 1. Take one of the large, red, deciduous stipules which cover the terminal bud of Ficus eldstica at the time when it is becoming transparent, a few days LABORATORY OUTLINES FOR GENERAL BOTANY. 85: before it is ready to fall. Examine immediately by holding the stipule with the inner side upon the stage of the microscope and examine with low and high power. Note the complex system of lactiferous ducts and the movement of the latex in them caused by its escape from the torn end of the stipule. At times the flow in the ducts appears to be very rapid. 2. Mount some of the latex and examine under high power. Note the spher- ical granules and draw. These are the rubber globules. 3. Why does the stipule become colored before it drops off from the bud? CIX. Pollen-tubes in Artificial Cultures. 1. From an opening anther take fresh pollen of Canna, Hyacinth, or Begonia and make cultures in the following solution: Agr OAC eS ale ac o.cy ature tony re Mire “Ciena ay te Oe Parts pean eres capaty Mer. raat kariave ahh cea) alka a DALtS Ce NAD AWALCL mene ep ass eee ate OL parts Heat the mixture over a water bath till the gelatin is dissolved. 2. To a cubic centimeter or two of this solution add an equal quantity of tap water and filter into a small covered dish. Put the pollen into the solution and also make hanging drop cultures, placing the slides into a moist chamber. 3. In 20 to 24 hours examine and draw several tubes representing successive stages of development. Note the rotation of the cytoplasm. 4. Treat with iodin solution and note the position of the nuclei. CX. Karyokinesis. Study the nuclear division in specially prepared slides of the root tips of Allium cépa L., the common onion. For a detailed study an oil immersion objective and compensating oculars are necessary, but much may be learned with the ordinary lenses. For staining use the fourth and fifth stains given in the appendix under “paraffin imbedding.” 1. Resting nucleus. Draw a cell some distance back of the tip where all the nuclei are in the resting condition. Represent the cell wall, the cytoplasm with vacuoles, and the nucleus. In the nucleus observe carefully the chromatin network with chromatin granules and the nucleoli. The nucleus is enclosed in the nuclear membrane. The lightly staining or hyaline substance in the nucleus, seen between the meshes of the chromatin network is called achromatin. 2. Prophase. (a) In the first stage of division the chromatin network is transformed into a continuous thread or spirem wound rather irregularly. At the same time the incept of the achromatic spindle appears forming two dome-shaped projections on opposite sides of the nucleus. This figure is known as the close mother skein. Find a suitable figure, draw and describe. (b) Later the looped mother skein is formed by the shortening and thick- ening of the continuous spirem which is thrown into a definite number of loops, the heads of which in typical cases point toward the two poles of the spindle. The nucleoli and nuclear membrane begin to disappear and the dome-shaped caps of the spindle become more pointed. Draw and describe. 3. Metaphase. (a) After the nuclear membrane disappears the spirem breaks into separate loops which are drawn into the equatorial plane with their heads toward the centre. At the same time the spindle continues to elongate. This. figure is known as the broken mother skein. Draw and describe. R6 LABORATORY OUTLINES FOR GENERAL BODANS. (b) When the chromosomes have come into the equatorial plane, there is a pause resulting from the seeming pull of the spindle fibers in opposite directions and the chromosomes are arranged in a very perfect star-shaped figure known as the mother star. Each chromosome has in the meantime commenced to split lon- gitudinally. This may be seen in the more advanced mother stars. Draw and describe. 4. Anaphase. (a) After longitundinal segmentation of the mother chromo- somes has taken place, the daughter chromosomes are gradually pulled apart, the separation beginning at the heads of the loops. This stage is called metakinesis. In very favorable sections, centrospheres or small round bodies may be seen at the poles of the spindle, also polar radiations, but these can only be studied favor- ably with an oil immersion lens. Draw and describe. (b) After the chromosomes have been completely separated they arrange themselves in star-shaped figures around the poles, while a central spindle of threads appears between the two stars. This is the daughter star stage. Draw and describe. d. Telophase. (a) The chromosomes being oriented around the poles, now begin to contract, becoming wavy in outline, and the free ends curve inward. The threads of the central spindle begin to thicken preparatory to the formation of the cell plate. Then the central spindle begins to bulge outward until it reaches the cell wall. Nucleoli begin to appear. In favorable figures the polar radiations are quite prominent and two centrospheres may be seen at the poles under a high objective. This stage is called the loose daughter skeim. Find a suitable figure, draw, and describe. (b) After the daughter cells are completely separated by the new cell wall the threads of the central spindle and the radiations around the poles disappear, and nuclear membranes appear around the daughter nuclei. The chromosomes begin to be transformed again into an expanding chromatin network. This is known as the close daughter skein. Select a suitable figure, draw and describe. 6. Make a series of diagrams showing the changes in the chromatin from one resting stage to another. Cx1. The Reduction Division. Study specially prepared slides of the ovularies of Lilium philadélphicum L., or the stamens of Lilium tigrinum Andr. The best preparations can probably be obtained from the stamens of Lilium tenwifolium Fisch. Delafield’s hematoxylin stain will bring out the chromatin well. The proper stages are obtained some time before the flower opens. In the ovule of Lilum the archesporial cell is transformed directly into the megasporocyte and this divides to form the first two cells of the embryosac, no true megaspores being formed. During this karyokinesis the chro- mosomes are reduced and undergo a qualitative division. Instead of Lilium the common Hyacinth may be used and will give excep- tionally good slides for studying and counting the eight bivalent chromosomes. Plant Hyacinth bulbs about October 22, in sawdust, and kill the flowers about November 1. The root-tips may be used for the vegetative karyokinesis. Stain in Delafield’s haematoxylin. 1. Under an oil immersion lens, study the early stages during which the chro- matin network is being transformed into a spirem. Draw and describe. 2. Draw the stage when the spirem shows a single row of chromatin granules. 3. Study the stage in which the chromatin granules are dividing preparatory to a longitudinal splitting of the linin thread. Draw. LABORATORY OUTLINES FOR GENERAL BOTANY. ; 87 4. Study the stage in which the spirem, after the chromatin granules have: divided, is twisting up into loops. There-are twelve of these loops which will break apart and form the twelve chromosomes; just half as many as in the previous divisions during the life of the sporophyte. Draw carefully and make a. diagram representing a loop and the arrangement of the chomatin granules. 5. Draw a chromosome just after the loops have broken apart, showing the: twisted spirem and the closed head end of the loop. These are bivalent chromo- somes made up of two univalents which are synaptic mates. 6. Draw a spindle in the mother star stage showing the fully developed chromosomes. Make a diagram illustrating the structure of a chromosome. Count. the chromosomes in a favorable cell. 7. Study the metakinesis stage when the chromosomes have partly untwisted. and appear like elongated bands on the spindle, just before they break at the: center. Draw. Make a diagram showing how the chromosome is attached to the spindle and how it divides. 8. Make a drawing of the early daughter star stage when the division of the: chromosomes is complete. 9. Make a series of diagrams showing the changes in the chromatin from the: early stage of division to the daughter skein stage. Compare the reduction division with the vegetative division where longitudinal splitting of the chromo-— somes takes place. CXII. Fluctuating Variability. One thousand soy beans (Soja max (L.) or other suitable seeds, all taken from successive plants until the number is procured, may be weighed and assorted into glass tubes, each tube containing the beans weighing within 25 milligrams of each other. If small beans are used, a 10 milligram interval may be employed; or if large beans are at hand, a 50 milligram interval in weighing will give satisfactory results. The tubes may be placed in a box or frame and one or a few sets, if properly managed, will be sufficient for the class. In small classes each student. may weigh a thousand beans, if suitable balances are at hand. 1. Count the beans in each tube and make a proper tabulation of the number for each weight. 2. Make a series of slender, contiguous rectangles with equal bases and by shading show diagramatically the difference in number of the various weights. of beans. 3. Plot a frequency curve showing the fluctuation of the 1000 beans. Could you state a principle relative to the abundance of the largest and smallest beans as. compared with those of intermediate weight? Look up Quetelet’s law. CXIII. Mendelian Principles of Heredity. Take pure white and pure red dent corn or flint corn and cross pollinate them —i. e. put pollen of red dent on the stigmas (silks) of white dent or the same for the flint corn if desired. Keep the silks covered properly to prevent the access of foreign pollen. Keep samples of the parent type of ears. Plant the hybrid corn and pollinate the hybrid plants among themselves. The hybrid corn will all have: grains with red pericarps (sporophyte character). Plant the red grain and in the next season there will be + white corn plants and ? red corn plants. The white: if kept separate will remain white pure, but 4 of the red will give pure red and 3 of it will give red and white again in the proportion of 3:1. It will be seen,. 2 88 | LABORATORY OUTLINES FOR GENERAL BOTANY. therefore, that } was pure red, } pure white, and } hybrid red. The type ears of the original parents and of the three succeeding generations are to be ar- ranged and labeled in proper order in a glass case or glass jars and the student is to note the striking hereditary results. Also have corn showing first generation hybrid ears with purple and white grains. 1. Note that in the first hybrid generation (F:) the red color only appears. The red is thus said to be dominant over the white. 2. Note that in the second hybrid generation (F:) pure white corn is again produced from the red. The white character is said to be recessive to the red. Note that there is no blending of the two characters red and white but that the white is developed as pure from the red. Note the ratio between the red and — white. Let R stand for red and w for white and if in the Fi generation eggs are produced having the R heredity and others having the w heredity and if the same is true for the sperms, make a diagram showing all possible combinations that could occur in fertilization of these two kinds of eggs and two kinds of sperms. Using the letters R and w as symbols, arrange the combinations on a checkerboard of the proper number of squares. 3. Note the results in the third generation (F;), (a) that the whites pro- duced only whites, (b) that 4 of the reds produced only reds, and (c) # of the reds, i. e. } of the F, generation, produced both reds and whites again in the ratio Ofuoml: 4. Make a diagram with braces showing the inheritance from the original pure parents to the third generation of offspring. 5). Note that these generations of corn illustrate the main facts of Mendel’s laws of heredity—namely, (a) dominance and recessiveness, (b) the lack of blending of certain hereditary factors or the characters resulting from their activity during development, and (c) the segregation of the factors and char- acters in definite ratios. 6. If white and purple endosperm characters are hybridized in corn could you explain, by remembering the facts in relation to the female gametophyte of the Anthophyta, why in the Fi generation of ears there would be both purple and white grains present on the same ear, while in the case of the red and white corn the ears (both in the Fi and F: generations) were always either completely white or red? 7. Ifa pure sweet corn ear is pollinated with pollen from both sweet corn and field corn (starch corn) all the silks that receive starch corn pollen from which fertilization results will show starchy endosperm in the grains, the starchy char- acter being dominant. This immediate effect of the pollen is called xenia and is the result of triple fusion and dominance. Examine such ears. Under what conditions could you tell that the ear had come from a pure stalk of sweet corn? APPENDIX. GENERAL METHODS IN BOTANICAL MICROTECHNIQUE. GENERAL METHOD WITH PARAFFIN IMBEDDING. In the following account all details are carefully stated, so that a beginner should be able, with little outside help, to carry the operations thru successfully. The methods employed in preparing plant tissues must be considerably different from those used in zoology, since we usually have to deal with a thick cellulose wall and a very delicate protoplasm in which are usually contained large vacuoles. filled with cell sap, besides numerous plastids and food contents, all of which tend to make it difficult to preserve and study the finer details of structure in plant cells and tissues. The object taken for a trial study may be some root tips of the common onion (Allium cepa), or pieces of the young ovularies of some species of lily, as Lilium longttiorum or L. philadelphicum, The root tips may be grown by placing an onion in a flower pot with moist sawdust, and keeping it for a few days where the roots will grow rapidly. The tips should be cut from one-half to three- fourths of a centimeter in length. The lily ovularies may be taken at various stages before and after the flowers open, and cut into transverse pieces from one-fourth to three-fourths of a centimeter long. 1. — KILLING AND FIXING. The first thing to do in beginning to prepare any plant tissue for permanent mounting is to kill and fix it in such a way that it will preserve the minute struc- tures as near the living condition as possible. A sharp knife or scalpel should always be used, and great care taken so as not to bruise or injure any of the cells. Killing Fluid.— The killing fluid is made up as follows: ie Cliromier cid. Fenestra. 0 .O> STams. eee Glacinl py NCeriCH NC ts ye tetas on OL CC. RMN ater eure on Ce aertee Var ore eek sot OO. Oee, Have the killing fluid in a 4 oz. (120 cc.) bottle with a common cork. Sixty cubic centimeters (2 oz.) of the fluid will be enough to kill one or two dozen objects the size of the root tips. The material must be perfectly fresh and put into the killing fluid as soon as cut. They will usually sink to the bottom after a short time, especially if they are shaken'a little from time to time. If much trouble ts. experienced in having the objects float on the surface of the killing fluid, they may first be immersed for a very brief moment in 95 per cent. alcohol, and immediately after dropped into the killing fluid. This will cause them to sink; or force them (89) ‘O00 LABORATORY OUTLINES FOR GENERAL BOTAN ye down with a plug of cotton. The objects must be kept in the killing fluid from twelve to twenty-four hours. The onion root tips should be left at least twelve hours; for larger objects, a proportionately longer time. 2.— WASHING. After the tissues have been thoroly killed and fixed, the next thing neces- sary is to wash out the acid. This may be done by pouring off the acid and filling the bottle with water, and changing from time to time. They should be washed in this way from one to four hours, depending on how often the water is changed. A better way, however, is to set up and use the apparatus described as a con- venient washing apparatus in this appendix. The water used for washing should ‘be rather pure. If this is not the case, distilled water had better be used. 3.— DEHYDRATING AND HARDENING. The next step is to remove all water from the tissues, and this must be done very gradually or the tissues will shrink and the protoplasmic contents of the cells will be distorted so that the preparations will be worthless. To remove the water successive grades of alcohol are used. During this process the objects may still be kept in the same bottle. The amount of each grade of alcohol should be sufficient to cover the objects well. The various grades of alcohol should be made up and kept in a special set of bottles. It is best not to use the alcohol more than once for this process. Carry them thru in the following order: Lali per cent se Aleololhe 2 see. mare igh ed atm hours. 2. Zo pet cent. Alcohols) = ins ee + room ome 3: i oon pem cent: “Alcohol so. 3. 2 ey tons emeanse A 50 per scent: Aleoholw:: Giro aes) cin 4 ato, opMerinss OO epEh Conky MIGOlON Nay aon ee ge hee ‘hours. The objects should be hardened in the 70 per cent. alcohol at least two days, and a longer period is generally better. They may be kept in 70 per cent. alcohol for several months without injury. 6. eSbipericent.