Showing papers in "Plant Physiology in 1972"

Obligatory reduction of ferric chelates in iron uptake by soybeans.

Abstract: The contrasting Fe2+ and Fe3+ chelating properties of the synthetic chelators ethylenediaminedi (o-hydroxyphenylacetate) (EDDHA) and 4,7-di(4-phenylsulfonate)-1, 10-phenanthroline (bathophenanthrolinedisulfonate) (BPDS) were used to determine the valence form of Fe absorbed by soybean roots supplied with Fe3+-chelates. EDDHA binds Fe3+ strongly, but Fe2+ weakly; BPDS binds Fe2+ strongly but Fe3+ weakly. Addition of an excess of BPDS to nutrient solutions containing Fe3+-chelates inhibited soybean Fe uptake-translocation by 99+%; [Fe(II) (BPDS)3]4− accumulated in the nutrient solution. The addition of EDDHA caused little or no inhibition. These results were observed with topped and intact soybeans. Thus, separation and absorption of Fe from Fe3+-chelates appear to require reduction of Fe3+-chelate to Fe2+-chelate at the root, with Fe2+ being the principal form of Fe absorbed by soybean.

A gas chromatographic method for the determination of aldose and uronic Acid constituents of plant cell wall polysaccharides.

Abstract: A major problem in determining the composition of plant cell wall polysaccharides has been the lack of a suitable method for accurately determining the amounts of galacturonic and glucuronic acids in such polymers A gas chromatographic method for aldose analysis has been extended to include uronic acids Cell wall polysaccharides are depolymerized by acid hydrolysis followed by treatment with a mixture of fungal polysaccharide-degrading enzymes The aldoses and uronic acids released by this treatment are then reduced with NaBH4 to alditols and aldonic acids, respectively The aldonic acids are separated from the alditols with Dowex-1 (acetate form) ion exchange resin, which binds the aldonic acids The alditols, which do not bind, are washed from the resin and then acetylated with acetic anhydride to form the alditol acetate derivatives The aldonic acids are eluted from the resin with HCl After the resin has been removed, the HCl solution of the aldonic acids is evaporated to dryness, converting the aldonic acids to aldonolactones The aldonolactones are reduced with NaBH4 to the corresponding alditols, dried and acetylated The resulting alditol acetate mixtures produced from the aldoses and those from the uronic acids are analyzed separately by gas chromatography This technique has been used to determine the changes in composition of Red Kidney bean (Phaseolus vulgaris) hypocotyl cell walls during growth, and to compare the cell wall polysaccharide compositions of several parts of bean plants Galacturonic acid is found to be a major component of all the cell wall polysaccharides examined

Salt responses of enzymes from species differing in salt tolerance

Abstract: Enzymes which are affected by the addition of inorganic salts during in vitro assay were extracted from salt-sensitive Phaseolus vulgaris, salt-tolerant Atriplex spongiosa, and Salicornia australis and tested for sensitivity to NaCl. In each case malate dehydrogenase, aspartate transaminase, glucose 6-phosphate dehydrogenase, and isocitrate dehydrogenase showed NaCl responses similar to those found for commercially available crystalline enzymes from other organisms. Enzymes extracted from plants grown in saline cultures showed no important changes in specific activity or salt sensitivity. Interaction of pH optima and NaCl concentrations suggests that enzymes may differ in the way they respond to salt treatment.

The Role of Protein Synthesis in the Senescence of Leaves: I. The Formation of Protease

Abstract: The senescence of oat leaves has been studied by following the loss of chlorophyll and protein and the increase of α-amino nitrogen, after detachment and darkening. Protein synthesis and the amounts of proteolytic enzymes in the leaves have been determined directly. The process of senescence is shown to be a sequential one in which protein synthesis,most probably the formation of a proteolytic enzyme with l-serine in its active center, is of prime importance. The evidence is as follows. Firstly, l-serine specifically enhances senescence, especially in presence of kinetin. Secondly, cycloheximide, which inhibits protein synthesis in other systems, delays senescence and prevents the serine enhancement. Although requiring higher concentrations, cycloheximide can be as effective as kinetin in inhibiting senescence. It is shown directly that cycloheximide prevents protein synthesis in oat leaves under the same conditions as when it prevents senescence. Thirdly, leaves have been shown to contain two proteinases, with pH optima at 3 and 7.5, whose activity increases during senescence, even though the total leaf protein is decreasing. The amounts of both these enzymes present after 3 days are clearly increased by serine, and are greatly decreased by cycloheximide or by kinetin. The role of kinetin in delaying senescence thus may rest on its ability to suppress protease formation.

Penetration of stomata by liquids: dependence on surface tension, wettability, and stomatal morphology.

Abstract: Wettability of the leaf surface, surface tension of the liquid, and stomatal morphology control penetration of stomata by liquids. The critical surface tension of the lower leaf surface of Zebrina purpusii Bruckn. was estimated to be 25 to 30 dyne cm−1. Liquids having a surface tension less than 30 dyne cm−1 gave zero contact angle on the leaf surface and infiltrated stomata spontaneously while liquids having a surface tension greater than 30 dyne cm−1 did not wet the leaf surface and failed to infiltrate stomata. Considering stomata as conical capillaries, we were able to show that with liquids giving a finite contact angle, infiltration depended solely on the relationship between the magnitude of the contact angle and the wall angle of the aperture. Generally, spontaneous infiltration of stomata will take place when the contact angle is smaller than the wall angle of the aperture wall. The degree of stomatal opening (4, 6, 8, or 10 μm) was of little importance. Cuticular ledges present at the entrance to the outer vestibule and between the inner vestibule and substomatal chamber resulted in very small if not zero wall angles, and thus played a major role in excluding water from the intercellular space of leaves. We show why the degree of stomatal opening cannot be assessed by observing spontaneous infiltration of stomata by organic liquids of low surface tension.

Nutrient Status and Mycorrhizal Enhancement of Water Transport in Soybean

Abstract: Mycorrhizal soybean (Glycine max L. Merr. var. Harosoy-63) plants have lower resistances to water transport than nonmy-corrhizal plants after 4.5 weeks of growth. Although resistances of whole plants differ by 40%, there were no differences in the resistances of stems plus leaves, indicating that the major effect of the mycorrhizae was to reduce the resistance of the roots. Since the fungitoxicant, p-chloronitrobenzene, had no effect on resistances to water transport, reduced resistances were probably not caused by a direct modification of the transport pathway by the fungus. Differences in resistance between mycorrhizal and nonmycorrhizal soybean were essentialy eliminated by the application of nutrients to the soil. Thus, lowered resistances of mycorrhizal roots growing in soil with low levels of nutrients probably resulted from the enhanced nutrient status of the plant brought about by the fungus. Mycorrhizal infection increased growth at both low and high nutrient levels.

Spherosome membranes: half unit-membranes.

Abstract: Spherosomes are bounded by unusual single-line "membranes" which measure 2 to 3.5 nanometers in width, contrasted to the well known tripartite unit-membranes which measure 6 to 8.5 nanometers in over-all thickness. Juxtaposed externally (from the side addressing the hyaloplasm), two spherosomal membranes adjoin to form a thicker single line, but apposed internally (the sides that contact stored lipid) two single-line membranes touch to form a tripartite structure resembling a unit-membrane. Morphologically, we interpret the single-line membranes of spherosomes as half unit-membranes whose polar surfaces face the hyaloplasm and whose lipoidal nonpolar surfaces contact internal storage lipid.Corroboration of this interpretation was shown biochemically by demonstrating the presence of membrane structural protein in peanut spherosomes. In addition, an immunological identity between membrane protein isolated from spherosomes of quiescent seeds and membrane protein extracted from the mitochondrial fraction of 10-day germinated seedlings was observed. We conclude that the atypical, single-line membranes bounding spherosomes are in fact biological membranes that correspond to half unit-membranes.

Four Universal Forms of Chlorophyll a

Abstract: We have matched the red absorption band measured at −196 C in a variety of chloroplast preparations with four major component curves representing forms of chlorophyll a having peaks at 661.6, 669.6, 677.1, and 683.7 nanometers. Chloroplast fractions enriched in one or the other of the two photochemical systems both contain these four major components, but system 1 preparations contain relatively more chlorophyll a 684. Chlorophyll a 677 and chlorophyll a 684 have greater bandwidths in system 1. Bands at longer wavelengths near 693 and 704 nanometers also often occur, but with far smaller heights than the above major bands. The longer wavelength bands are more common in system 1 than in system 2. In system 1 the half-widths of the four major bands in typical spectra average 11.3, 10.0, 10.3, and 10.8 nanometers while in system 2 they are 11.6, 9.8, 9.4, and 9.6 nanometers. Some spectra with sharper and some with wider bands were found, but the wavelengths were identical.

Cytokinins in seedling roots of pea.

Abstract: The natural occurrence of cytokinins existing both in a free form and as a constituent of transfer RNA was examined in serial segments of young seedling roots of pea. Purified ethanol extracts of root apices were resolved into four factors capable of inducing soybean callus tissue proliferation. The most active factor was identified as zeatin or some closely related compound; it produced polyploid divisions and tracheary element differentiation when tested on cultured pea root segments. The terminal 0- to 1-millimeter root tip contained 43 to 44 times more free cytokinin on a fresh weight or a per cell basis than the next 1- to 5-millimeter root segment. Extracts of more proximal segments behind the tip contained no measurable free cytokinin. Acid hydrolysates of transfer RNA exhibited reproducible cytokinin activity. Bioassays revealed that the predominant amounts of free cytokinin and that present in transfer RNA were restricted to the extreme root tip. There was approximately 27 times more free cytokinin than the amount detected in transfer RNA in root apices.

Abscisic Acid and stomatal regulation.

Abstract: The closure of stomata by abscisic acid was examined in several species of plants through measurements of CO(2) and H(2)O exchange by the leaf. The onset of closure was very rapid, beginning at 3 minutes from the time of abscisic acid application to the cut base of the leaf of corn, or at 8 or 9 minutes for bean, Rumex and sugarbeet; rose leaves were relatively slow at 32 minutes. The timing and the concentration of abscisic acid needed to cause closure were related to the amounts of endogenous abscisic acid in the leaf. Closure was obtained in bean leaves with 8.9 picomoles/cm(2). (+)-Abscisic acid had approximately twice the activity of the racemic material. The methyl ester of abscisic acid was inactive, and trans-abscisic acid was likewise inactive. The effects of stress on levels of endogenous abscisic acid, and the ability of very small amounts of abscisic acid to cause rapid closure suggests that stomatal control is a regulatory function of this hormone.

The Requirement for Sodium as a Micronutrient by Species Having the C4 Dicarboxylic Photosynthetic Pathway

Abstract: Six species having characteristics of plants with the C(4) dicarboxylic photosynthetic pathway, Echinochloa utilis L Ohwi et Yabuno (Japanese millet), Cynodon dactylon L (Bermuda grass), Kyllinga brevifolia Rottb, Amaranthus tricolor L cv Early splendour, Kochia childsii Hort, and Portulaca grandiflora Hook (rose moss), responded decisively to 01 milliequivalent per liter NaCl supplied to their culture solutions initially containing less than 008 microequivalent per liter Na Chlorosis and necrosis occurred in leaves of plants not receiving sodium Portulaca failed to set flower in the sodium-deficient cultures Under similar conditions Poa pratensis L (Kentucky blue grass) having characteristics of the C(3) photosynthetic pathway made normal growth and did not respond to the addition of sodium It is concluded from these results and previously reported work that sodium is generally essential for species having the C(4) pathway but not for species with the C(3) pathway

Polyribosomes from Peas: An Improved Method for Their Isolation in the Absence of Ribonuclease Inhibitors

Abstract: Profiles of polyribosomes were obtained from etiolated stem segments of Pisum sativum L. var. Alaska isolated in various buffers. Tissue homogenized in a medium containing 0.2 m tris-HCl, pH 8.5, 0.2 m sucrose, 30 mm MgCl2, and 60 mm KCl yielded polyribosomes exhibiting far less degradation than tissue homogenized in conventional media containing tris-HCl at lower ionic strength and pH. A further decrease in degradation was found when polyribosomes were sedimented through a sucrose pad buffered at pH 8.5 prior to centrifugation. Increased separation was obtained using heavy (125-500 mg/ml), linear sucrose gradients. Using these techniques, messenger RNA species bearing up to 12 ribosomes (dodecamers) were resolved, with messenger RNA chains bearing 9 ribosomes (nonamers) being the most abundant (having the highest absorption peak). The data presented suggest that buffer of high ionic strength and high pH was more effective in preventing degradation of polyribosomes than was diethyl pyrocarbonate and, furthermore, that ratios involving large polyribosomes (hexamers and larger) were more accurate indices of degradation than were ratios involving total polyribosomes.

Canopy and Seasonal Profiles of Nitrate Reductase in Soybeans (Glycine max L. Merr.)

Abstract: Nitrate reductase activity of soybeans ( Glycine max L. Merr.) was evaluated in soil plots and outdoor hydroponic gravel culture systems throughout the growing season. Nitrate reductase profiles within the plant canopy were also established. Mean activity per gram fresh weight per hour of the entire plant canopy was highest in the seedling stage while total activity (activity per gram fresh weight per hour times the total leaf weight) reached a maximum when plants were in the full bloom to midpod fill stage. Nitrate reductase activity per gram fresh weight per hour was highest in the uppermost leaf just prior to full expansion and declined with leaf position lower in the canopy. Total nitrate reductase activity per leaf was also highest in the upper-most fully expanded leaf during early growth stages. Maximum total activity shifted to leaf positions lower in the plant canopy with later growth stages. Nitrate reductase activity of soybeans grown in hydroponic systems was significantly higher than activity of adjacent soil grown plants at later growth stages, which suggested that under normal field conditions the potential for nitrate utilization may not be realized. Nitrate reductase activity per gram fresh weight per hour and nitrate content were positively correlated over the growing season with plants grown in either soil or solution culture. Computations based upon the nitrate reductase assay of plants grown in hydroponics indicated that from 1.7 to 1.8 grams N could have been supplied to the plant via the nitrate reductase process. The harvested seed contained 1.1 to 1.2 grams N per plant. Thus, based on previous estimates of approximately 32% of the final N distribution being in the vegetative plant parts, the estimated input of reduced nitrogen via the enzyme assay was in agreement with the actual N accumulation. The amount of calculated N 2 -fixation by nodules per season with plants grown in hydroponics was less than 2% of the computed nitrate reduced via leaf nitrate reductase. Thus, the level of nitrate in the nutrient solution appeared to be quite inhibitory to N 2 -fixation.

Anaerobic Accumulation of γ-Aminobutyric Acid and Alanine in Radish Leaves (Raphanus sativus, L.)

Abstract: In leaves, the anaerobic accumulation of alanine was accompanied by a loss of aspartate, and these changes preceded gamma-aminobutyrate accumulation and glutamate loss. Changes in keto acid content did not appear to be the cause of amino acid changes. Accumulation of gamma-aminobutyrate was due to acceleration of glutamate decarboxylation and arrest of gamma-aminobutyrate transamination. Changes in enzyme content did not explain the changes in reaction rates in vivo. Most of the aspartate may be converted anaerobically to alanine via oxalacetate and pyruvate.

Studies on cytokinin production by Rhizobium.

Abstract: Cytokinin was released into the medium by cultures of both Rhizobium japonicum and R. leguminosarum. Calculations show that the amount of cytokinin released during the logarithmic phase of growth by R. japonicum would be sufficient to initiate the cortical cell divisions necessary to form a root nodule. The substance released by R. japonicum was identified as a zeatin-like compound on the basis of paper chromatography in four solvent systems. Two solvents clearly separated the rhizobial product from N6-Δ2-isopentenyladenine and its ribonucleoside. The predominant intracellular cytokinin found in both enzymatic hydrolysates of sRNA and alkaline hydrolysates of total RNA also was similar to zeatin.

The Correlated Appearance of Prolamellar Bodies, Protochlorophyll(ide) Species, and the Shibata Shift during Development of Bean Etioplasts in the Dark.

Abstract: The structure and physiology of the etioplast was investigated in developing primary leaves of 3- to 9-day-old dark-grown bean ( Phaseolus vulgaris L. var. Red Kidney) seedlings. Increase in total protochlorophyll(ide) content followed that of leaf fresh weight. In 3- to 4-day-old bean leaves more than 50% of the protochlorophyll(ide) is in the form of protochlorophyll(ide) 628, which is nontransformable by light. Most of the transformable pigment is protochlorophyll(ide) 635, with smaller amounts of protochlorophyll(ide) 650. During leaf development from the 3rd to the 7th day phototransformable protochlorophyll(ide) with an absorbance maximum at 650 nm accumulates faster than nontransformable protochlorophyll(ide) or protochlorophyll(ide) 635. This increase in protochlorophyll(ide) 650 is correlated with the formation and enlargement of prolamellar bodies. The transformable protochlorophyll(ide) is converted by light directly to chlorophyll(ide) 672 in young leaves which do not yet have prolamellar bodies, and chlorophyll(ide) 672 may arise largely from the protochlorophyll(ide) 635. In older leaves the protochlorophyll(ide), largely protochlorophyll(ide) 650, is converted to chlorophyll(ide) 683, and a Shibata shift results in a change in the wavelength of absorption to 672 nm. The increase in protochlorophyll(ide) 650, the formation of prolamellar bodies, and the presence of the Shibata shift appear to be closely correlated. A model is briefly presented to provide a unified interpretation of these findings, including certain similarities between dark-grown Euglena cells and 3- to 4-day-old etiolated bean leaves.

Volatile metabolites controlling germination in buried weed seeds.

Abstract: Velvetleaf (Abutilon theophrasti Medic), morning glory (Ipomoea purpurea [L.] Roth), and wild mustard (Brassica kaber [D.C.] L. C. Wheeler) seeds exhibited decreased germination with increased planting depth in soil. Flushing the soil for 2 minutes each day with air overcame the inhibition. A sealed in vitro system was used to sample the volatile components produced by weed seeds. Inhibition of seed germination was accompanied by decreased O2 levels and production of volatile metabolites identified as acetaldehyde, ethanol, and acetone. The effectiveness of these compounds in reducing germination was dependent on O2 levels.

Auxin-induced Changes in Avena Coleoptile Cell Wall Composition.

Abstract: Sugar and uronic acid residues were derived from wall polysaccharides of oat (Avena sativa, var. Victory) coleoptiles by means of 2 N trifluoroacetic acid, 72% sulfuric acid, or enzymic hydrolysis. The products of hydrolysis were reduced and acetylated to form alditol acetates which were analyzed using gas chromatography. Time-course studies of auxin-promoted changes in various wall fractions indicate that when exogenous glucose was available, increases in certain wall constituents paralleled increases in length. However, under conditions where exogenous glucose was not available, and where wall synthesis was limited, such correlations with growth were not apparent. Under these latter conditions total wall weight initially increased slightly, then decreased. These changes in weight were the net of increases in cellulose and some noncellulosic constituents and a decrease of over 75% in noncellulosic glucose. When coleoptile sections were preincubated without exogenous glucose for 8 hours to deplete endogenous wall precursors and subsequently treated with auxin, there were no detectable increases in wall weight. There was instead an auxin-promoted decrease in wall weight, and this decrease paralleled a decrease in noncellulosic glucose. There were no significant changes in other wall components. The auxin-promoted decreases in noncellulosic glucose are interpreted as a possible step in the mechanism of growth.

A Cell Wall-degrading Endopolygalacturonase Secreted by Colletotrichum lindemuthianum

Abstract: Cultures of Colletotrichum lindemuthianum (Saccardo and Magnus) Scribner have been induced to secrete an endopolygalacturonase (polygalacturonide glycanohydrolase EC3.2. 1.15). This enzyme has been brought to a high state of purity by ion exchange, gel filtration, and agarose affinity chromatography. The enzyme has optimal activity at pH 5, has an apparent molecular weight as determined by gel filtration of about 70,000, and prefers polygalacturonic acid to pectin as its substrate. The enzyme, while hydrolyzing only 1% of the glycosidic bonds, reduces the viscosity of a polygalacturonic solution by 50%. Nevertheless, the initial as well as the final products of polygalacturonic acid hydrolysis are predominantly tri- and digalacturonic acid and, to a lesser extent, monogalacturonic acid. The purified enzyme catalyzes the removal of about 80% of the galacturonic acid residues of cell walls isolated from suspension-cultured sycamore cells (Acer pseudoplatanus) as well as from the walls isolated from 8-day-old Red Kidney bean (Phaseolus vulgaris) hypocotyls.

Effect of Ethylene on Cell Division and Deoxyribonucleic Acid Synthesis in Pisum sativum

Abstract: Ethylene and supraoptimal levels of 2,4-dichlorophenoxyacetic acid inhibit the growth of the apical hook region of etiolated Pisum sativum (var. Alaska) seedlings by stopping almost all cell divisions. Cells are prevented from entering prophase. The hormones also retard cell division in intact root tips and completely stop the process in lateral buds. The latter inhibition is reversed partially by benzyl adenine. In root tips and the stem plumular and subhook regions, ethylene inhibits DNA synthesis. The magnitude of this inhibition is correlated with the degree of repression of cell division in meristematic tissue, suggesting that the effect on cell division results from a lack of DNA synthesis. Ethylene inhibits cell division within a few hours with a dose-response curve similar to that for most other actions of the gas. Experiments with seedlings grown under hypobaric conditions suggest that the gas naturally controls plumular expansion and cell division in the apical region.

The Cupric Ion as an Inhibitor of Photosynthetic Electron Transport in Isolated Chloroplasts

Abstract: Strong inhibition of uncoupled photosynthetic electron transport by Cu(2+) in isolated spinach chloroplasts was observed by measuring changes in O(2) concentration in the reaction medium. Inhibition was dependent not only on the concentration of the inhibitor, but also on the ratio of chlorophyll to inhibitor. Binding of Cu(2+) to the chloroplast membranes resulted in removal of Cu(2+) from solution. When chloroplasts were exposed to preincubation in light, there was increased inhibition as a result of Cu(2+) binding to inhibitory sites. Preincubation in the dark resulted in Cu(2+) binding to noninhibitory sites and decreased inhibition. The degree of inhibition was lower at low light intensities than at high light intensities.When the photosystems were assayed separately, photosystem I was more resistant to inhibition than photosystem II. The most sensitive site to the inhibitor was the oxidizing side of photosystem II.

Salt responses of carboxylation enzymes from species differing in salt tolerance.

Abstract: This paper reports effects of salts on in vitro activity of phosphoenolpyruvate carboxylase and ribulose-1,5-diphosphate carboxylase, isolated from species differing in salt tolerance.Inhibition of phosphoenolpyruvate carboxylase by the inorganic salts KCl, NaCl, and Na(4)SO(4) depended on the source of the enzyme. Phosphoenolpyruvate carboxylase isolated from leaves of C(4) plants was extremely sensitive to inorganic salts, whereas the enzyme extracted from roots of C(4) plants or from both shoots and roots of C(3) plants was much less sensitive. Ribulose-1,5-diphosphate carboxylase was less salt-sensitive than the phosphoenolpyruvate carboxylases. Differences in salt sensitivity of carboxylases were observed over a wide pH range. The results suggest substantial physical-chemical differences between phosphoenolpyruvate carboxylases functioning in photosynthesis and in CO(2) dark fixation.Among C(4) species, phosphoenolpyruvate carboxylase from halophytic species was more salt-sensitive than that from a salt-sensitive species. This anomaly, between in vitro response of enzymes and growth response of the plants, is briefly discussed.

Developmental studies on microbodies in wheat leaves : I. Conditions influencing enzyme development.

Abstract: Catalase, glycolate oxidase, and hydroxypyruvate reductase, enzymes which are located in the microbodies of leaves, show different developmental patterns in the shoots of wheat seedlings. Catalase and hydroxypyruvate reductase are already present in the shoots of ungerminated seeds. Glycolate oxidase appears later. All three enzymes develop in the dark, but glycolate oxidase and hydroxypyruvate reductase have only low activities. On exposure of the seedlings to continuous white light (14.8 x 10(3) ergs cm(-2) sec(-1)), the activity of catalase is doubled, and glycolate oxidase and hydroxypyruvate reductase activities increase by 4- to 7-fold. Under a higher light intensity, the activities of all three enzymes are considerably further increased. The activities of other enzymes (cytochrome oxidase, fumarase, glucose-6-phosphate dehydrogenase) are unchanged or only slightly influenced by light. After transfer of etiolated seedlings to white light, the induced increase of total catalase activity shows a much longer lag-phase than that of glycolate oxidase and hydroxypyruvate reductase. It is concluded that the light-induced increases of the microbody enzymes are due to enzyme synthesis. The light effect on the microbody enzymes is independent of chlorophyll formation or the concomitant development of functional chloroplasts. Short repeated light exposures which do not lead to greening are very effective. High activities of glycolate oxidase and hydroxypyruvate reductase develop in the presence of 3-amino-1,2,4-triazole which blocks chloroplast development. The effect of light is not exerted through induced glycolate formation and appears instead to be photomorphogenetic in character.In senescing leaves excised from the plants decreases in activity of glycolate oxidase, and hydroxypyruvate reductase follow with some delay the decrease in chlorophyll content. The activity of catalase, however, is maintained at high levels, especially when the detached shoots are kept in light.

Biochemical Changes in the Rice Grain during Germination

Abstract: Changes in the content of starch, protein, and RNA and in the activity of their hydrolases in the rice endosperm (Oryza sativa L., variety IR8) were determined during the first week of germination without added nutrient both in the dark and in the light. Changes were generally more rapid in the dark than in the light. Oxygen uptake and RNase activity started to increase and the root protruded on the second day, followed by the coleoptile on the third day, and the primary leaf on the fourth day. ATP level was at a maximum on the fourth day. The activity of amylases and R enzyme increased progressively, but that of phosphorylase tended to decrease during starch degradation. A new alpha amylase isozyme band appeared during germination. Glucose was the major product of starch degradation. Sucrose, maltose, maltotriose, raffinose, and fructose were also detected. Protease activity reached a maximum on the fifth or sixth day and closely paralleled the increase in soluble amino N and soluble protein.In embryoless seed halves with 0.12 muM gibberellin As, peak protease activity occurred in 2.5 days and peak alpha amylase activity on the fifth day of incubation. The production of alpha amylase, protease, and R enzyme was inhibited by 40 muM cycloheximide, but only alpha amylase and R enzyme were inhibited by 20 mug/ml actinomycin D.

Migration of Calcium and Its Role in the Regulation of Seismonasty in the Motor Cell of Mimosa pudica L.

Abstract: Volume and conformational changes of the contractile tannin vacuoles of the abaxial motor cells of the primary pulvinus of Mimosa pudica L. parallel the seismonastic leaf movement. Since such changes in cells and organelles of animal systems are often regulated by calcium, we studied Ca 2+ movement in the motor cells and tissue. By fixation with Lillie9s neutral buffered formalin, followed by staining with alizarin red sulfate (ARS), calcium was localized in the tannin vacuoles of the motor cells of the primary pulvinus. After treatment with ethylenediaminetetraacetate, 8-hydroxyquinoline, and several other calcium-complexing or extracting agents, the color reaction due to alizarin red sulfonate was no longer present. By using an analytical method, it was shown that the effluent from stimulated pulvini has significantly more Ca 2+ than that from unstimulated controls. Ten millimolar LaCl 3 inhibits recovery of the tannin vacuole in vivo in 10 mm CaCl 2 or in distilled water. Quantitative data obtained by microspectrophotometry demonstrated calcium migration during the bending movement of the primary pulvinus. In the adaxial motor cells a small amount of calcium migrates from the tannin vacuole, and calcium on the cell wall moves to the central vacuole. In the abaxial half, a large amount of calcium from the tannin vacuole moves to the central vacuole of the motor cell. It is probable that the calcium binds to the microfibrillar contents of the central vacuole. These observations support the contention that Ca 2+ migrates between the surface of the tannin vacuole and the inside of the central vacuole. The recovery and maintenance of the tannin vacuole in the spherical form may play a role in maintaining turgor in the motor cells of the abaxial half of the primary pulvinus of Mimosa .

Interrelationships of Ethylene and Abscisic Acid in the Control of Rose Petal Senescence

Abstract: The role of abscisic acid and ethylene in the senescence of rose petals cv. Golden-Wave was examined. A rise in ethylene evolution, followed by an increase in the level of abscisic acid was observed. The presence of abscisic acid in rose petals was established, using different chromatography systems, several bioassays, and immunoassay. External application of ethylene accelerated senescence and induced a rise in endogenous abscisic acid-like activity. Application of abscisic acid promoted senescence, but suppressed ethylene production. The data suggest that the participation of these two hormones in the control of senescence is via the same pathway. The possibility of interrelationship between abscisic acid and ethylene was tested and experimental evidence in favor of this hypothesis is presented. It was suggested that ethylene affects senescence in rose petals by inducing an increase in abscisic acid activity, which in turn may control ethylene evolution, via a feedback mechanism.

Ethylene Evolution from Cucumber Plants as Related to Sex Expression

Abstract: Ethylene evolved from monoecious and gynoecious cucumber (Cucumis sativus) plants grown under short and long day conditions was determined. More ethylene was evolved from floral buds and apices bearing buds than from whole seedlings of comparable weight. More ethylene also was evolved from apices of the gynoecious than from those of the monoecious type. Furthermore, quantities evolved from female buds were greater than from male ones and plants grown under short day conditions which promote femaleness evolved more ethylene than those grown under long day conditions. The data suggest that ethylene participates in the endogenous regulation of sex expression by promoting femaleness.

Synthesis and turnover of nitrate reductase in corn roots.

Abstract: The induction and reinduction of nitrate reductase in root tip or mature root sections show essentially a similar pattern: a lag, a period of rapid increase in enzyme activity and finally a period of relatively minor change. Both inductions are sensitive to 6-methylpurine and cycloheximide. Kinetic studies with 6-methylpurine suggest that the half-life of the messenger RNA for nitrate reductase in both sections is about 20 minutes. The rate of decay of nitrate reductase activity induced by transfer to a nitrate-free medium is slower in root tips (t½ = 3 hours) than in mature root sections (t½ = 2 hours). The enzyme from mature root sections is also less stable to mild heat treatments (27 C; 40 C) than the enzyme from root tip sections. The results indicate that factors regulating enzyme turnover show important changes as root cells mature and may be significant in determining steady state levels of the enzyme.

Effects of medium composition and carbon dioxide on circadian conidiation in neurospora.

Abstract: Efforts to significantly perturb the timing mechanism, and thus the period, of the rhythm responsible for circadian conidiation in bd, a strain of Neurospora crassa, by altering the medium composition have been unsuccessful. Various salt solutions, sugars, and amino acids do, however, have pronounced effects on growth and conidiation, and thus on the expression and persistence of rhythmicity.Aeration stimulates conidiation in growth-tube cultures, thereby allowing experiments which demonstrate that nearly all strains and species of Neurospora are capable of circadian conidiation. These results extend to Neurospora the generalization that physiological and developmental regulation in all eukaryotic organisms has a circadian component. Aeration also increases the persistence of circadian conidiation suggesting that the damping of rhythmicity previously observed on certain media represents the cessation of conidiation, rather than the stopping of a timing mechanism.Aeration is apparently effective in that it maintains CO(2) levels in growth-tube cultures below a critical concentration. Carbon dioxide was shown to inhibit conidiation in both wildtype and bd strains, with the latter being about 200 times more resistant than the former.