NUCLEAR MIGRATION IN GERMINATING SPORES OF ONOCLEA SENSIBILIS: THE PATH AND KINETICS OF MOVEMENT

Nuclear migration was observed in individual germinating spores of Onoclea sensibilis from the onset of movement to the completion of mitosis. About 16 hr after the initiation of germination, the nucleus migrated from its initial position in the center of the spore to the proximal side. It then appeared to migrate along the raphe to one end of the spore where an asymmetric division occurred. The average velocity of migration was measured at 0.256 ± 0.065 μm/min. During migration the nucleus underwent changes in shape. No migrations or movement other than that by the nucleus were observed.     

STORAGE PRODUCTS IN SPORES OF ONOCLEA SENSIBILIS L

Lipids and proteins in substantial quantities are present as storage products in spores of Onoclea sensibilis. They fill the sparce spore cytoplasm and ultrastructurally are indistinguishable from reserve materials observed in storage tissue of higher plants. Hydrolysis of both products is correlated with early stages in spore germination.     

CO-REQUIREMENT FOR CALCIUM AND POTASSIUM IN THE GERMINATION OF SPORES OF THE FERN ONOCLEA SENSIBILIS

Spores of Onoclea sensibilis L. do not germinate on distilled H₂O if they are pretreated for sufficient time with dilute NaClO solution. However, spores will germinate, after NaClO pretreatment, on a simple mineral medium containing the major and trace elements. Complete germination after pretreatment also is obtained on a solution containing only Ca²⁺ and K⁺ as the cations, but neither ion by itself is sufficient. Rb⁺, but not Li⁺ or Na⁺, can replace K⁺. Hypochlorite-treated spores do not require the continuous presence of Ca²⁺ and K⁺ to germinate; exposure during the first 4 hr of culture, with the remainder of the time on distilled H₂O, is sufficient. Extraction of spores with ethylene glycol bis(aminoethyl ether) tetraacetic acid [EGTA] makes their germination dependent on Ca²⁺, as reported by other workers, but it does not produce a co-requirement for K⁺. Colorimetric analysis with arsenazo III confirms that Ca²⁺ is extracted from Onoclea spores by NaClO. Extractable Ca²⁺ amounts to about 78 nmol/mg spore dry wt. Of this amount, 31% is contained in the perispore. The perispore comprises 13% of the total spore dry wt.     

NUCLEAR MIGRATION AND ASYMMETRIC CELL DIVISION IN ONOCLEA SENSIBILIS SPORES: AN ULTRASTRUCTURAL AND CYTOCHEMICAL STUDY

A method of preparation for electron microscopy of fern spores in early stages of germination is presented. The cytochemistry and fine structure of Onoclea spores during the early stages of germination are described. The cytoplasm of the hydrated spore is filled with lipid droplets, protein granules and chloroplasts. During the early stages of development ribosomes and mitochondria increase in the area surrounding the central nucleus, and a new peripheral wall forms around the protoplast. Microtubules and large, branching mitochondria are associated with the nucleus during migration from its original central position in the spore to the proximal face and then to one end of the spore. There is no morphological polarization of cytoplasmic organelles of the spore before migration of the nucleus.     

ANALYSIS OF STARCH ACCUMULATION AND GERMINATION IN ONOCLEA SPORES

Previous studies have shown that low fluences of light accelerate starch accumulation and enhance germination of Onoclea spores. Fluence response curves for induction of starch accumulation were compared with fluence response curves for enhancement of germination in order to determine if the two photoresponses in Onoclea spores have a common photoreceptor. Fluence response curves indicate that both responses were proportional to the log of the fluence and that the relative fluence efficiencies of the four wavelength regions tested were similar for induction of both germination and starch accumulation. Red (600–720 nm) irradiation was the most efficient, while green (500–600 nm), blue (400–520 nm), and far-red (720–900 nm) irradiations showed a decreasing order of efficiency for induction of the responses. A correlation coefficient between the amount of starch accumulated as a result of red irradiation and the final percent germination was calculated to be 0.964. These results support the hypothesis that a common photoreceptor mediates both photoinduced germination and starch accumulation. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) inhibits photosynthesis by blocking the flow of electrons from Photosystem II to Photosystem I. At 0.1 mM DCMU failed to inhibit both photoinduced starch acumulation and germination. This result and the greater efficiency of red than blue light, the low fluence functional for induction, and the fluence dependency argue against the participation of photosynthesis in photoinduced starch accumulation. A similar conclusion has been previously drawn for photoenhancement of Onoclea spore germination. Additionally, the effects 0.01–1.0 mm cycloheximide and 100 μl/l ethylene on photoinduced starch accumulation were investigated. Neither agent inhibited starch accumulation, whereas both substances inhibited germination 70–90% when applied at a time coincidental with the period of rapid starch accumulation. These results indicate that the photoinduction of starch accumulation does not have an ethylene sensitive stage nor does it require protein synthesis as does photoenhancement of germination of Onoclea spores.     

THE EFFECT OF DIFFERENT LIGHT CONDITIONS AND SUCROSE ON THE GROWTH AND DEVELOPMENT OF THE GAMETOPHYTE OF THE FERN,ONOCLEA SENSIBILIS

Miller , J. H. (Yale U., New Haven, Conn.), and P. M. Miller . The effect of different light conditions and sucrose on the growth and development of the gametophyte of the fern, Onoclea sensibilis. Amer. Jour. Bot. 48(2): 154–159. Illus. 1961.—Gametophytes grow rapidly under the highest intensity white fluorescent light used, 400 ft.-c. Growth is correspondingly less at 220 and 28 ft.-c., and at the lowest light intensity the gametophytes are markedly longer than wide. One percent sucrose added to the medium reverses the growth limitation imposed by low light intensity and produces a morphology comparable to that found under higher intensities. Sucrose does not sustain heterotrophic growth in total darkness, but permits growth if the gametophytes are given low dosages of red light. The effect of red light is reversible by far-red illumination.     

BIOCHEMISTRY OF FERN SPORE PROTEINS: GLOBULIN STORAGE PROTEINS IN ONOCLEA SENSIBILIS AND OSMUNDA CINNAMOMEA

Onoclea sensibilis was found to contain globulin storage proteins of 2.0S and 11.3S. These globulins, comparable in size and subunit composition to the spore storage proteins characterized in Matteuccia struthiopteris (Templeman, DeMaggio, and Stetler, 1987), declined during imbibition and the initial stages of spore germination. Osmunda cinnamomea, a fern that is only distantly related to Matteuccia, also contained globulin proteins, but these had S values of 5.5 and 11.3. SDS-PAGE analysis revealed extensive differences in banding patterns of the 11.3S protein between Onoclea and Osmunda. This work indicates that while globulin proteins are important storage materials in a variety of ferns they exhibit considerable molecular heterogeneity. The observed heterogeneity in the globulin proteins may be a useful tool to explore evolutionary relationships in the ferns.     

ULTRASTRUCTURAL FEATURES OF THE DIVIDING MEIOCYTE OF ONOCLEA SENSIBILIS

The ultrastructure of the anaphase of the second meiotic division of the meiocyte of Onoclea sensibilis is characterized by a compact plate of mitochondria without the usual double limiting membrane and with single-layered cristae continuous from one side of the mitochondrion to the other. Amyloplasts occur proximal to the plate and lipid droplets are randomly distributed throughout the cell.     

GROWTH PATTERNS OF PEA SEEDLINGS IN DARKNESS AND IN RED AND WHITE LIGHT

Thomson , Betty F., and Pauline Monz Miller . (Connecticut Coll., New London.) Growth patterns of pea seedlings in darkness and in red and white light. Amer. Jour. Bot. 48(3): 256–261. Illus. 1961.—Seedlings of peas were grown in vermiculite at 22°C. and exposed 16 hr. daily to red or white light or kept in darkness. Others were grown in soil in the greenhouse. Samples harvested daily to 16 days were dissected, the length of each internode and leaf measured and the total number of leaves and leaf primordia counted. The form of the stem apex and youngest primordia and interrodes is the same in light as in darkness. Leaf production is accelerated very slightly and the growth of leaves and internodes is decidedly accelerated by light. Leaf-leaf, leaf-internode and internode-internode correlations indicate that the morphogenetic effect of light is limited to later stages of organ growth. Dry weight is consumed more rapidly in light than in darkness, probably because of more rapid growth and slightly greater amounts of respiring tissue in light-grown plants.     

ORIENTATION BY OPPOSED BEAMS OF LIGHT

Computations of the effective angular inclination (H) of the photoreceptive surfaces of the two sides, based upon measurements of orientation angles under the action of beams of light directly opposed or crossing at right angles, show that with larvae of Calliphora and of Lucillia H declines as the total illumination decreases (i.e., as the angle of orientation away from the more intense light increases). H is greater with the two lights opposed at 180°; this may be due to the difference in refraction. For the more sharply pointed larvae of Lucillia, H is less than half as great as in Calliphora.     

ORIENTATION AND LOCUS OF TROPIC PHOTORECEPTOR MOLECULES IN SPORES OF BOTRYTIS AND OSMUNDA

Study of the tropic responses of Botrytis cinerea and Osmunda cinnamomea spores to blue light shows the photoreceptor molecules to be highly dichroic and oriented: in Botrytis their axes of maximum absorption lie perpendicular to the nearby cell surface; in Osmunda, parallel. The chief evidence lies in a comparison of their responses to plane polarized light—both germinate parallel to the vibration planes (defined by the axis of vibration of the electric vector and the axis of light propagation)—with those to partial illumination with unpolarized light: Botrytis grows from its brighter part; Osmunda, from its darker. The degree of orientation produced by polarized light corresponds, at high intensities, to that produced by the imposition of such large (about 100 per cent) intensity differences across a cell as to preclude all alternatives to oriented dichroic receptors. The photoreceptors of the Botrytis spore lie within the cell wall's inner half. The chief evidence lies in the component of its tropic responses to polarized light within the vibration plane: germination peaks about 10° off the vibration axis. This deviation arises from focusing which is effective only in the wall's inner half. At high intensities, anomalies appear in Botrytis which are interpreted as "centering," i.e., a tendency toward growth from the center of two or more equally illuminated points of a cell rather than from one of them.     

LEAF DEVELOPMENT OF PHASEOLUS VULGARIS L. IN LIGHT AND IN DARKNESS

Development of the primary bean leaf in the dark and under continuous white light was studied during 14 days after sowing. The increase in surface area of the blade is the result of a number of sequential processes. Both in the darkness and under illumination, leaf growth is characterized by an initial cell enlargement followed by intensive cell division. Cell division in etiolated leaves continues for one day longer than in illuminated ones, but it proceeds at a slower rate. Mature leaves grown under white light undergo a phase of cell enlargement after cell division has stopped. This increases their surface area up to 800 times when compared with the blade area of the embryo. This enlargement phase is almost absent in dark-grown seedlings. Consequently the blade area of etiolated leaves is only 50 times that of the embryonic state. Thus light appears to have a dual effect on leaf development: it activates cell division and induces cell expansion.     

PHOTIC ORIENTATION BY TWO POINT-SOURCES OF LIGHT

General formulæ are derived for the orientation of phototropic organisms in a field illuminated by two point-sources of light in the same horizontal plane. It is shown that formulæ previously found may all be derived as special cases of these general equations.     

THE EFFECT OF AUXIN AND GUANINE ON CELL EXPANSION AND CELL DIVISION IN THE GAMETOPHYTE OF THE FERN,ONOCLEA SENSIBILIS

Miller , J. H. (Yale U., New Haven, Conn.) The effect of auxin and guanine on cell expansion and cell division in the gametophyte of the fern, Onoclea sensibilis. Amer. Jour. Bot. 48(9): 816–819. Illus. 1961.—Auxin and guanine promote cell expansion in 0. sensibilis gametophytes. The optimum concentration of auxin for total expansion is 10^−-5 M, but the optimum for elongation is 10^−-6 M. Above this concentration the cells expanded isodiametrically. Guanine is active at higher concentrations than auxin. Increasing concentrations of auxin progressively inhibit red light-induced cell division, while guanine has no effect on cell division. Neither kinetin nor adenine promotes cell expansion or cell division.