STUDIES ON COMPONENTS OF RESPIRATORY SYSTEM OF BEGONIA LEAVES

Investigations were made of the properties of diaphorase, cytochromec reductases, cytochrome c oxidase, and other components ofelectron transfer system in various fractions of leaf homogenateof Begonia semperflorens.

1. All the fractions tested showed the existence of cytochromec oxidase, succinic- and reduced diphosphopyridine nucleotide-cytochromec reductases, and diaphorase. Activities of these enzymes werefound to be associated mainly with the particulate fractions.The particulate fractions showed, in particular, a capacityof reducing oxidized cytochrome c with fumarate, malate, -ketoglutarate,ß-hydroxy-butyrate, and citrate.
2. Optimum pH foroxidation of cytochrome c by the particulatefractions was foundto be 5.5, while that for reduction was7.2.
3. The activityof cytochrome c reductase was partially suppressedby malonate.Partial inhibition of cytochrome c oxidase wascaused by azideand cyanide, the inhibitory effects observedbeing strongerwith particulate fractions than with solublefractions.

(Received August 11, 1962; )     

PHYSIOLOGICAL STUDIES ON THE VERTICILLIUM WILT DISEASE OF TOMATO

The water loss per unit leaf area of tomato plants was decreased after inoculation with Verticillium albo-atrum. When diseased plants began to wilt water loss temporarily increased, but then rapidly decreased to become less than that of healthy plants grown under conditions of adequate or restricted water supply.
The transpiration of excised leaves from plants grown with a restricted water supply was reduced, but not so severely as that of comparable leaves from infected plants. Water loss from leaves on infected plants was reduced irrespective of any blocking of the petiolar xylem.
The rate of water loss from turgid leaf disks on mannitol solutions, and the rate of water uptake of leaf disks on water was similar for disks cut from wilting or turgid leaves of diseased plants or healthy plants grown with an adequate or restricted water supply.
Disease or poor water supply reduced leaf growth but had no effect on the rate of leaf initiation. Although the density of stomata was higher on leaves of diseased plants the stomatal area was less than on healthy plants.
The resistance to water flow in diseased stems was high and was correlated with vessel blockage. About half the blocked vessels contained hyphae. The severity and localization of symptoms in inoculated plants growing on susceptible or resistant rootstocks was directly related to the extent of invasion by the pathogen and to vessel blockage.
Experiments on the wilting activity of cell-free filtrates from cultures of the pathogen in vitro indicated that it produced a stable substance, not an enzyme, that caused wilting in cut shoots by blocking the end of the stem. It is suggested that an increasing internal water shortage causes major symptoms of disease.     

STUDIES ON PROTEIN SYNTHESIS IN TOMATO COTYLEDONS AND LEAVES. I. PROTEIN SYNTHESIS AND TURNOVER IN INTACT COTYLEDONS AND LEAVES

1. Details of aseptic culture of virus-free tomato seedlings usedin comparative in vivo and in vitro studies on protein synthesisare described.
2. Developmental changes in the levels of DNA,RNA, protein andchlorophyll content of seedling cotyledonsand leaves were recorded,and are related to protein synthesis.
3. Incorporation of isotopically labeled carbon into proteinwasfollowed both by photosynthetic uptake of ¹⁴CO₂ and by theuptakeof ¹⁴C-amino acids through the roots.
4. A marked stimulationby light of ¹⁴C uptake was observed, andthe higher rate of¹⁴C incorporation from ¹⁴CO₂ than from ¹⁴C-aminoacids intothe protein fraction is discussed in relation tothe pathwaysof protein synthesis in tomato leaves, and alsowith regardto protein turnover.

¹Present address: Dept. of Horticultural Science, Universityof Wisconsin, U.S.A.     

STUDIES ON THE RE-ASSIMILATION OF RESPIRATORY CO2 IN ILLUMINATED LEAVES

Experiments were performed, using rice, barley and Hydrangealeaves, to examine the re-assimilation of respiratory ¹⁴CO₂while photosynthesis is going on in an open air flow system. It was found that the leaves which had assimilated ¹⁴CO₂ beforehandevolved, when kept under photosynthesizing conditions, threeto four tenths (variable according to plant species and externalconditions) of the amount of ¹⁴CO₂ to be produced in the dark.Such an incomplete re-utilization of ¹⁴CO₂ was observed alsoin spinach leaf homogenate as well as in the leaves which hadpreviously absorbed ¹⁴C-glucose. The ¹⁴CO₂ output in rice leaves was found to be acceleratedby the light of high intensity. A possibility of light stimulationon the respiration was suggested. (Received October 7, 1961; )     

STUDIES ON THE PHYSIOLOGICAL EFFECTS OF NON-POLAR-POLAR ORGANIC ELECTROLYTES : I. THE INFLUENCE UPON THE RESTING POTENTIAL OF FROG MUSCLE

1. The commonly used detergents have a poisonous effect, which is due to the non-polar-polar configuration of their organic anion. The non-polar organophilic half of the ion is built up by a long chain of alkyl radicals (8 to 18 carbon atoms), the polar hydrophilic half by a sulfonate or sulfate. If brought into contact with the organic surface membrane of a cell, this structure, due to the strong attachment of the alkyl chains to its surface, and due to the pull of the hydrophilic part towards the surrounding water, is subjected to a heavy stress terminating in tearing to pieces the membrane (by denaturing and loosening the membrane components; bacteriolysis, cytolysis). Correspondingly, with frog muscle, one end of which has been treated with the detergent solution, an irreversible negative injury potential is produced. 2. Applying, instead, the compounds bearing short chains of alkyl radicals (1 to 6 carbon atoms), producing less stress on the membrane and correspondingly a slighter derangement of its architecture, a reversible positive resting potential appears. This is interpreted to be the effect of the non-polar part of the anion, which, due to its surface activity, intrudes into the pores of the membranes, notwithstanding the negative charge of their walls. 3. The short chained detergents seem to be replaceable by various organic "semidetergents," the organophilic behavior of their anion being represented by a slight chemical affinity (NH₂), the hydrophilic by the effect of a carboxyl group (COO) instead of sulfate or sulfonate. The effect of the semidetergents on muscle is a positive reversible potential. Their physiological significance may be visualized as a functional activation.     

STUDIES ON PROTEIN SYNTHESIS IN TOMATO COTYLEDONS AND LEAVES. II. INTERMEDIATE STAGES OF PROTEIN SYNTHESIS

1. The activities of RNase, ATPase and proteolytic enzymes inextractsof tomato leaves have been investigated with regardto detrimentaleffects on protein synthesis. The use of coppersulphate asan inhibitor of RNase is described.
2. The effectof microbial contamination on amino acid uptake hasbeen studiedin relationship with cell-free systems, and thehigh activitynoted underlines the necessity for aseptic procedures.
3. Methodsfor the preparation of cell-free systems capable ofamino acidactivation and incorporation into protein in an asepticenvironmentare discussed.

¹Present address: Dept. of Horticultural Science, Universityof Wisconsin, Madison, Wisconsin, U.S.A.     

GRAFTING AND ROOTING OF LEAVES OF GUAREA (MELIACEAE): EXPERIMENTAL STUDIES ON LEAF AUTONOMY

The pinnately compound, indeterminate leaves of G. glabra and G. guidonia were air layered, detached from their original shoots, and grown on their own adventitious root systems for up to 58 mo and 26 mo, respectively. The detached leaves grew in the same indeterminate manner and reached sizes similar to attached leaves. Although detached leaves grew autonomously, they never produced shoot buds. Leaves of both species were grafted onto their own stems and cut free of their original leaf bases. Leaf scions survived and grew for up to 29 mo and 20 mo, respectively, similar to ungrafted leaves. Axillary branches were grafted onto subtending leaves. Branch scions grew on their leaf stocks for over 30 mo and 24 mo, respectively, after being cut free from the branch bases. Secondary growth of the leaf axis (petiole) was promoted, and vascular tissues of leaf and branch axes were continuous. However, the unlignified basal region of the leaf, including the abscission zone, remained unchanged after grafting. The results indicate that proximity of roots and bypassing the abscission zone did not enhance leaf longevity or pinna production. The presence of a growing branch on a leaf did not modify the structure of the abscission zone, which suggests that the zone is strongly committed or developmentally fixed.     

乳酸脱氨酶DAB－环己烷反胶束溶液中的活性

研究了兔肌乳酸脱氢酶Ｍ４（ＬＤＨ）在十二胺丁酸盐（ＤＡＢ）－环己反烷胶束溶液中的催化活性。发现ＬＤＨ在ＤＡＢ反胶束中的催化转换数（Ｋｃａｔ）同水溶液中的相近。ＬＤＨ在ＤＡＢ反胶束中的活性随增溶水量的增加而增加,随ＤＡＢ浓度的增加而降低,文中还提出了ＬＤＨ在ＤＡＮ反胶束中的增溶方式。     

乳酸脱氨酶DAB－环己烷反胶束溶液中的活性

研究了兔肌乳酸脱氢酶Ｍ４（ＬＤＨ）在十二胺丁酸盐（ＤＡＢ）－环己反烷胶束溶液中的催化活性。发现ＬＤＨ在ＤＡＢ反胶束中的催化转换数（Ｋｃａｔ）同水溶液中的相近,ＬＤＨ在ＤＡＢ反胶束中的活性随增溶水量的增加而增加,随ＤＡＢ浓度的增加而降低,文中还提出了ＬＤＨ在ＤＡＢ反胶束中的增溶方式。     

STUDIES OF NITRATE TRANSPORT BY MARINE PHYTOPLANKTON USING 36Cl-ClO3− AS A TRANSPORT ANALOGUE. I. PHYSIOLOGICAL FINDINGS1

Transport of a nitrate analogue, ³⁶Cl-ClO₃⁻, was examined in two diatoms, Skeletonema costatum (Greve.) Cleve and Nitzschia closterium (Ehrenb) W. Sm. A dinoflagellate, Gonyaulax polyedra did not transport ClO₃⁻. Transport of ³⁶Cl-ClO₃⁻, by diatoms appeared to be active and showed saturation kinetics. The data were fitted by Michaelis-Menten equation at all but the lowest chlorate concentrations (where plots of S vs. v showed a slight concave bend). Affinity of cells for nitrate was considerably higher than for chlorate. The K_i for nitrate inhibition of chlorate transport was calculated assuming competitive inhibition. Light had little or no effect on chlorate transport. Pulse-chase experiments demonstrated that (1) ClO₃⁻ (hence NO₃⁻) was stored in two intracellular compartments of equal size, (2) internal ClO₃⁻ was exchangeable with external ClO₃⁻ (rates of efflux and influx were measured), and (3) efflux of intracellular ClO₃⁻ showed transient states following a chase of ClO₃⁻ or NO₃⁻ which stabilized after 10–20 min. Transport of chlorate was a function of growth phase.     

STUDIES OF NITRATE TRANSPORT BY MARINE PHYTOPLANKTON USING 36Cl-ClO3− AS A TRANSPORT ANALOGUE. I. PHYSIOLOGICAL FINDINGS1

Transport of a nitrate analogue, ³⁶Cl-ClO₃⁻, was examined in two diatoms, Skeletonema costatum (Greve.) Cleve and Nitzschia closterium (Ehrenb) W. Sm. A dinoflagellate, Gonyaulax polyedra did not transport ClO₃⁻. Transport of ³⁶Cl-ClO₃⁻ by diatoms appeared to be active and showed saturation kinetics. The data were fitted by Michaelis-Menten equation at all but the lowest chlorate concentrations (where plots of S vs. v showed a slight concave bend). Affinity of cells for nitrate was considerably higher than for chlorate. The K_i for nitrate inhibition of chlorate transport was calculated assuming competitive inhibition. Light had little or no effect on chlorate transport. Pulse-chase experiments demonstrated that (1) ClO₃⁻ (hence NO₃⁻) was stored in two intracellular compartments of equal size, (2) internal ClO₃⁻ was exchangeable with external ClO₃⁻ (rates of efflux and influx were measured), and (3) efflux of intracellular ClO₃⁻ showed transient states following a chase of ClO₃⁻ or NO₃⁻ which stabilized after 10–20 min. Transport of chlorate was a function of growth phase.     

利用抗生素喷施黄金葛叶片减轻冻害的研究

用91μg/mL氨苄青霉素、91μg/mL硫酸链霉素混合对黄金葛（Scoindapsus aureum)叶生喷施,第15d下叶片,于－19℃下冷冻5min,结果表明：水处理叶片较未处理的对照叶片冻斑数目减少75％,冻斑厚度不变,冻斑均从叶片边缘开始。药品处理组冷冻5min未见冻斑;冷冻8min后,冻斑较对照减少95％,分布于叶片边缘。表明抗生素能明显减轻叶片的冻害。     

土壤杆菌固氮生理特性研究

对根癌土壤杆菌C58/pGV3850菌株的固氮生理特性研究结果表明,该菌具有自生固氮活性,其固氮活性的最适pH为8.0,温度为30℃.固氮活性在对数生长后期(14h)最高,延缓期和静止期乙炔还原活性较低.该菌株好氧,通过氧呼吸和吸氢酶的作用达到避氧固氮.当通入氧气超过呼吸耗氧时,对固氮活性产生抑制作用.在培养过程中增加NH_4~+浓度,固氮活性会降低,当达到15mmol/L时完全丧失固氮活性,表现NH_4~+阻遏固氮酶蛋白合成.在乙炔还原测定系统中加入NH_4~+并不影响乙炔还原反应,说明没有NH_4~+关闭现象.培养过程中加入MSX(2.5mg/ml)能解除10mmol/L NH_4~+对固氮酶合成的阻遏作用.固定的游离氮不能以NH_4~+形式分泌于胞外.固氮过程中放出大量氢气,培养16小时产氢量达65μmol H_2/mg蛋白.在限碳条件下(0.2%蔗糖)其吸氢酶活力可达520nmol H_2·ml~(-1)·h~(-1).