Activity Changes in Selected Enzymes from Soybean Leaves Following Ozone Exposure

Soybeans [Glycine max(L.) Merr.] were harvested at various time periods after a 2-h exposure to either 0 or 0.5 μ1/1 ozone to determine the effects of ozone on selected enzymes. Carbohydrate metabolism was modified by a depression of glyceraldehyde 3-phaosphate dehydrogenase and an activation of glucose 6-phosphate dehydrogenase. Ozone did not alter the levels of RNase, protease, acid phosphatase or esterase as might be expected if ozone enhanced leaf senescence. The activities of phenylalanine ammonia lyase, polyphenol oxidase and peroxidase were initially depressed and then stimulated following the ozone exposure. The reactions of soybeans to an acute ozone stress were more nearly akin to those elicited in response to other stresses than to the process of senescence.     

昼间亚高温下番茄叶中糖含量与蔗糖代谢相关酶的活性日变化

研究番茄品种‘辽园多丽’幼苗在昼间35℃亚高温条件下叶中糖含量及蔗糖代谢相关酶活性日变化的结果表明,昼间亚高温处理后的幼苗叶中果糖、葡萄糖和淀粉含量下降,蔗糖含量升高。与蔗糖代谢相关的酶活性有明显的昼夜节律性变化,转化酶、蔗糖合成酶呈现昼间活性低、夜间活性高的节律性,而蔗糖磷酸合成酶活性在进入夜间时立刻升高。35℃昼间亚高温处理后的幼苗叶中,转化酶活性下降,蔗糖合成酶活性明显升高,蔗糖磷酸合成酶活性则略有升高。     

Changes in Activities of Antioxidant Enzymes in Sunflower Leaves of Different Ages

Role of superoxide dismutase isozymes and other antioxidant enzymes was studied in relation to leaf age in sunflower (Helianthus annuus L. cv. ACC 1508) at pre-flowering and grain filling stages. Relative water content (RWC) did not change much in leaves of different age and at the two stages. Protein content declined continuously from the youngest to the oldest leaf, while chlorophyll (Chl) and carotenoids (Car) contents increased down to 7^th/9^th leaf and declined in subsequent older leaves. Protein, Chl and Car contents were higher at pre-flowering than at seed filling stage. Superoxide dismutase (SOD), its isozymes, and ascorbate peroxidase (APO) and catalase (CAT) activities were highest in the 9^th leaf and declined in subsequent older leaves. SOD and APO activities were higher at seed filling, except in oldest senescent (13^th, 15^th) leaves. Among SOD isozymes, Cu/Zn-SOD and Mn-SOD activities accounted for most of the total SOD, and only marginal activity was observed for Fe-SOD. Peroxidase activity increased from youngest to the oldest leaf at pre-flowering stage and down to 13^th leaf at seed filling stage. This revised version was published online in July 2006 with corrections to the Cover Date.     

Activities of Antioxidant Enzymes during Senescence of Prunus Armeniaca Leaves

During the period of senescence of apricot leaves changes in photosynthetic pigment contents and in the activities of some antioxidant enzymes (superoxide dismutase, catalase, peroxidase and ascorbate peroxidase) were analysed. Significant changes in pigment contents were, in most cases, correlated with changes in activities of the antioxidant enzymes. Modifications in superoxide dismutase and catalase isoform patterns were also observed during the progression of senescence. Both enzyme activities and isoenzyme patterns proved to be genotype-dependent.     

Changes in Activities of Enzymes of Nitrogen Metabolism in Seedcoats and Cotyledons during Embryo Development in Pea Seeds

In the seedcoats of developing pea seeds, the maximal activities of asparaginase (EC 3.5.1.1) and aspartate: α-ketoglutarate aminotransferase (EC 2.6.1.1) are attained early in development, before the embryo has expanded to fill the embryo sac. These two enzyme activities could account for the early absence of asparagine and aspartate from the fluid secreted by the seedcoats into the embryo sac.     

The Development of Activities of Some Enzymes Concerned with Glycollate Metabolism in Greening Bean Leaves

The activities of phosphoglycollate phosphatase (EC 3.1.3.18 [EC] ),glycollate oxidase (EC 1.1.3.1 [EC] .). catalase (EC 1.11.1.6 [EC] ), theperoxisomal NADH-glyoxylate reductase (EC 1.1.1.26 [EC] ) which isconsidered to function as a hydroxypyruvate reductase in theperoxisomes, and the chloro-plastic NADPH-dependent glyoxylatereductaae, have been measured in extracts prepared from 14-d-olddark-grown bean leaves during the course of their greening inresponse to exposure to continuous illumination. All of theenzymes were found in the dark-grown leaves and on a per-leafbasis the activities increased from 6- to 12-fold with the exceptionof a 2–3-fold increase of NADPH-dependent glyoxylate reductaseduring 96-h greening, while the activities either remained constantor declined during similar periods in darkness. Initial lagperiods were evident before the illumination-induced increasesin enzyme activities. As D-threo-chloramphenicol did not affectthe increase in activity of any of these enzymes it would appearthat the increases were in no way dependent on protein synthesisby 70S ribosomes, or on the development of photosynthetic activity.     

Effect of Low Temperature on the Protein Metabolism of Wheat Leaves

The effect of low temperature on the protein metabolism of wheat primary leaves was examined. In seedlings transferred from 25 to 5 °C, total soluble protein accumulation, in vivo protein synthesis and breakdown, in vitro protein breakdown, and SDS-PAGE profiles of proteinases in gelatine-containing gels were analysed. Leaf protein content increased within a 7-d period (70 % over the initial value) in plants exposed to 5 °C. The fast protein accumulation observed on days 0 – 2 was mainly attributed to a decreased breakdown. In further days, parallelly to a slowdown in the rate of protein accumulation, the leaf proteolytic activity increased. The incubation temperature also had an influence on the proteolytic activity: Q ₁₀ values for the 15 – 5 °C range were 80 – 200 % higher than those observed for the 25 – 15 °C range. On the other hand, the in vivo protein synthesis capacity, at either 25 or 55 °C, was not significantly modified in cold-treated plants. In addition to the enhanced activities of two serine-proteinases (previously found in control plants by SDS-PAGE analysis), cold-treated plants displayed a new proteinase, which had not been detected so far.     

Activity of Carbon Metabolism Enzymes in Wheat Plants Treated with Kartolin-4 and Exposed to Water Stress

Enzymatic activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) (EC 4.1.1.39), phospho(enol)pyruvate carboxylase (EC 4.1.1.31), NAD malate dehydrogenase (EC 1.1.1.37), and NADP glyceraldehyde phosphate dehydrogenase complex including phosphoglycerate kinase (EC 2.7.2.3) and glyceraldehyde phosphate dehydrogenase (EC 1.2.1.13) were comparatively assayed in wheat seedlings of the cultivar Lyutestsens 758 grown under normal conditions, water deficiency conditions, and subsequent rehydration. Water stress was found to decrease the activity of all enzymes tested, the effect being most pronounced in the case of Rubisco. The content of Rubisco in wheat plants exposed to water deficiency was reduced less significantly than the activity of the enzyme. Pretreatment of plant seeds with kartolin-4 (o-isopropyl-N-2-hydroxyethyl carbamate), a preparation with cytokinin activity, reduced the dehydration-induced inhibition of enzymatic activity. Upon a subsequent rehydration, kartolin-4 facilitated rapid recovery of the photosynthetic activity, the process being based on the kartolin-induced stimulation of reparation reactions. Under conditions of water stress, a partial decrease in the activity of carbon metabolism enzymes in vitrowas accompanied by complete inhibition of photosynthesis in vivo, perhaps, as a result of an abrupt increase in the stomatal resistance.     

Hardly Increased Oxidative Stress After Exposure to Low Temperature in Chilling-Acclimated and Non-Acclimated Maize Leaves

Abstract: Seedlings of Zea mays L. were grown at optimal (25 °C) and suboptimal (15 °C) temperature and then exposed to severe chilling temperature (6 °C) at their growth light intensity (450 ìmol quanta m⁻² s⁻¹) for 4 d. Photosynthetic parameters, hydrogen peroxide, antioxidant contents, and activity of scavenging enzymes were investigated before, during, and after chilling stress. This stress caused a stronger reduction in photosynthetic activity, maximum quantum efficiency of photosystem II primary photochemistry ( F _v/ F _m), and catalase activity in plants which had been grown at 25 °C rather than at 15 °C. Maize plants grown at suboptimal temperature de-epoxidized their xanthophyll cycle pool to a greater extent and exhibited a faster recovery from chilling stress than plants which had not been acclimated to chilling. Antioxidant content, activity of scavenging enzymes, with the exception of catalase, hydrogen peroxide formation, and the size of the xanthophyll cycle pool were hardly affected by chilling stress. However, chilling induced a temporary increase in the glutathione content and triggered the synthesis of á-tocopherol during the phase of recovery at 25 °C. The results indicate that leaves respond to chilling stress by down-regulation of photosystem II accompanied by de-epoxidation of the xanthophyll cycle pool, probably to prevent enhanced formation of superoxide radicals at photosystem I and, consequently, other reactive oxygen species.     

Spore Germination and Carbon Metabolism in Fusarium solani V. Changes in Anaerobic Metabolism and Related Enzyme Activities during Development

Macroconidia of Fusarium solani f. phascoli have no detectable capacity to respire glucose anaerobically; germinated spores and mycelium, on the other hand, ferment glucose, although slowly.

Extracts of ungerminated spores contain hexokinase, phosphohexoisomerase, phosphofructokinase, aldolase, triose phosphate dehydrogenase, triose phosphate isomerase, phosphoglyceric kinase, enolase, phosphoglyceric mutase, pyruvate kinase, and pyruvate decarboxylase. It follows, therefore, that the appearance of fermentative capacity during spore germination cannot be ascribed to the de novo synthesis of any of these enzymes.

During germination and mycelial development the specific activity of all of the enzymes named except phosphohexoisomerase and aldolase increases 2- to 8-fold. Specific activity of all of the enzymes is substantially higher than the fermentative capacity of intact cells, i.e., none is limiting to anaerobic respiration.

The enzymatic assay data are consistent with a conclusion reached earlier on the basis of studies of aerobic glucose metabolism, that the process of germination involves an acceleration of pre-existing metabolic systems rather than an appearance of new pathways.

     

超高产杂交稻剑叶中C4途径酶活性和稳定碳同位素分异作用的变化

以超高产杂交水稻(Oryza sativa L.)"培矮64S/E32"和多年来大面积推广的杂交稻"汕优63"为材料,研究孕穗后剑叶中C4途径酶和对稳定碳同位素分异作用的变化.结果表明,籽粒灌浆期(移栽后68～75 d)的两个品种剑叶中NADP-MDH活性最高,随后下降;超高产杂交水稻"培矮64S/E32"的NADP-MDH的活性明显高于"汕优63";PEPCase和NADP-ME活性在黄熟期之前的叶片中持续上升.不同生育期的叶片与籽粒的△1aC值相近(19.49‰～19.82‰),在成熟期时较高.超高产水稻"培矮64S/E32"叶片的平均△13C值比"汕优63"高0.43‰.     

超高产杂交稻剑叶中C4途径酶活性和稳定碳同位素分异作用的变化

以超高产杂交水稻(Oryza sativa L．)“培矮64S／E32”和多年来大面积推广的杂交稻“汕优63”为材料,研究孕穗后剑叶中C4途径酶和对稳定碳同位素分异作用的变化。结果表明,籽粒灌浆期(移栽后68～75d)的两个品种剑叶中NADP—MDH活性最高,随后下降;超高产杂交水稻“培矮64S／E32”的NADP-MDH的活性明显高于“汕优63”;PEPcase和NADP—ME活性在黄熟期之前的叶片中持续上升。不同生育期的叶片与籽粒的△^13C值相近(19．49‰～19．82‰),在成熟期时较高。超高产水稻“培矮64S／E32”叶片的平均△^13C值比“汕优63”高0．43‰。     

Comparative Studies of Enzymes Related to Serine Metabolism in Higher Plants

THE FOLLOWING ENZYMES RELATED TO SERINE METABOLISM IN HIGHER PLANTS HAVE BEEN INVESTIGATED: 1) d-3-phosphoglycerate dehydrogenase, 2) phosphohydroxypyruvate:l-glutamate transaminase, 3) d-glycerate dehydrogenase, and 4) hydroxypyruvate:l-alanine transaminase. Comparative studies on the distribution of the 2 dehydrogenases in seeds and leaves from various plants revealed that d-3-phosphoglycerate dehydrogenase is widely distributed in seeds in contrast to d-glycerate dehydrogenase, which is either absent or present at low levels, and that the reverse pattern is observed in green leaves.The levels of activity of the 4 enzymes listed above were followed in different tissues of the developing pea (Pisum sativum, var. Alaska). In the leaf, from the tenth to seventeenth day of germination, the specific activity of d-glycerate dehydrogenase increased markedly and was much higher than d-3-phosphoglycerate dehydrogenase which remained relatively constant during this time period. Etiolation resulted in a decrease in d-glycerate dehydrogenase and an increase in d-3-phosphoglycerate dehydrogenase activities. In apical meristem, on the other hand, the level of d-3-phosphoglycerate dehydrogenase exceeded that of d-glycerate dehydrogenase at all time periods studied. Low and decreasing levels of both dehydrogenases were found in epicotyl and cotyledon. The specific activities of the 2 transaminases remained relatively constant during development in both leaf and apical meristem. In general, however, the levels of phosphohydroxypyruvate:l-glutamate transaminase were comparable to those of d-3-phosphoglycerate dehydrogenase in a given tissue as were those for hydroxypyruvate: l-alanine transaminase and d-glycerate dehydrogenase.     

Source--sink Relationships and Carbon Metabolism in Tomato Leaves 2. Carbohydrate Pools and Catabolic Enzymes

Levels of the major carbohydrates (hexoses, inositol, sucroseand starch) and activities of the enzymes (invertase, amylaseand starch phosphorylase) were assayed in the leaves and stemsof tomato plants. The transport system of the plants was simplifiedto one leaf and one associated truss either retained or removed.The experiment was carried out at two light intensities, 80and 20 W m^–2. Truss removal immediately increased thelevels of hexoses and starch in leaves under high light, butthose of sucrose and inositol were relatively unaffected withina day. Three days after truss removal the levels of sucrose,hexose and starch had reached a maximum. Under low light, leafsugar levels were lower and did not increase following trussremoval. The stem appeared to act as a substitute sink in thoseplants where the truss had been removed. Invertase and amylase activities were not significantly affectedby the treatments, but loss of starch phosphorylase activityduring the experiment was prevented by truss removal. Tomato, source-sink relationships, carbon, carbohydrate     

Response of Rice Leaves to Low Temperature I. Changes in Basic Biochemical Parameters

When the seedlings of two rice cultivars, IR8 (low-temperaturesensitive) and Somewake (low-temperature tolerant), were exposedto a low temperature of 15?C, the normal increase in the chlorophyllcontent of the developing 4th leaf blade completely ceased whileincreases in protein content continued at a low rate in bothcultivars. Analysis of soluble and insoluble proteins in the4th leaf blade of IR8 by SDS-polyacrylamide gel electrophoresisrevealed that synthesis of RuBP carboxylase and of several thylakoidproteins responsible for photosynthetic electron transport andphotophosphorylation was greatly inhibited at the low temperature.It was also found that increases in the activities of some enzymesof the Calvin cycle, such as RuBP carboxylase, fructose bisphosphataseand NADP-glyceraldehyde-3-P dehydrogenase, as well as of catalase,were specifically inhibited during growth at the low temperature.These results suggest that the synthesis of intracellular components,in particular of key proteins required for photosynthesis, isspecifically susceptible to low-temperature stress during developmentof rice leaves. (Received June 21, 1989; Accepted January 11, 1990)     

低温胁迫对建兰叶片内源多胺含量的影响

以素心建兰为材料,研究低温胁迫(5℃)对建兰内源多胺含量的影响。结果表明,在低温胁迫下建兰生长期的叶片多胺(PAs)总量和亚精胺(Spd)、腐胺(Put)含量都表现为先升后降的变化,精胺(Spm)含量则是下降然后上升最后稳定在比原来含量更高的水平上,说明有可能在低温胁迫下建兰通过调节内源多胺的总量和不同多胺种类的比例来抵御低温对生理的破坏作用;开花期的建兰叶片因受低温胁迫的影响未能合成足够的亚精胺(Spd),从而抑制了建兰的开花。     

Characteristics and Activity Changes of Proteolytic Enzymes in Apple Leaves during Autumnal Senescence

At least four different proteinases are present in senescing apple leaves (Malus domestica Borkh. cv. Golden Delicious) as determined by their pH optima, substrate specificity, and their reactivity to proteinase inhibitors. An enzyme active at pH 4.5 to 5.0 appears to be a sulfhydryl-dependent (iodoacetamide and phenylmercuric acetate-sensitive) endoproteinase, and degradation of the large subunit of ribulose bisphosphate carboxylase was observed only with this enzyme. It is tentatively concluded that this endoproteinase is responsible for the breakdown of ribulose bisphosphate carboxylase in vivo. However, the presence of more than one endoproteinase in apple leaves is suggested by the broad range of pH optima of the SH-dependent enzyme. Another enzyme active at pH 6.0 appears to be a carboxypeptidase, and was sensitive to phenylmethylsulfonylfluoride. This enzyme showed a strong hydrolytic activity against carbobenzoxyphenylalanylalanine. A sulfhydryl-dependent aminopeptidase and a second hydroxyl-dependent carboxypeptidase were active at pH 7.5

Total autolytic activity (the sulfhydryl-dependent endoproteinase) as measured by the disappearance of proteins decreased during the period of protein decline. Evidence is presented that the measured proteinase activity can be dependent on assay methods and substrates. While the disappearance of protein measures most of endo-type activity, the ninhydrin assay appears to measure exo-type activity preferentially.

     

夜间低温胁迫对番茄叶片活性氧代谢及AsA-GSH循环的影响

以番茄品种‘辽园多丽’为试材,利用人工气候室模拟设施生产中的夜间低温胁迫环境,研究9℃和6℃夜低温对番茄叶片活性氧代谢和AsA-GSH循环的影响。结果显示:9℃和6℃夜间低温胁迫3～9d可诱导番茄叶片中超氧阴离子(O2.-)产生速率、过氧化氢(H2O2)和丙二醛(MDA)含量上升;抑制过氧化物酶(POD)、过氧化氢酶(CAT)的活性,增加超氧化物歧化酶(SOD)和AsA-GSH循环中抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)、谷胱甘肽还原酶(GR)的活性,并提高还原型抗坏血酸(AsA)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)的含量。研究表明,在夜间低温胁迫过程中,增加的番茄叶片中SOD活性和AsA-GSH循环清除活性氧的能力并未与氧还原的速率一致,从而导致番茄叶片中活性氧的累积,使细胞膜系统受到一定破坏,在6℃处理的植物中尤为明显。     

石刁柏体细胞胚发生过程中多胺代谢酶和过氧化氢清除酶的活性变化

石刁柏体细胞胚发生过程中精氨酸脱羧酶(ADC)活性变化趋势与内源腐胺的含量变化相同,多胺氧化酶(PAO)活性变化幅度较小.外源腐胺对ADC活性的影响与不作外源腐胺处理的差异不显著性,但促进体细胞胚发生前期的PAO活性,过氧化物酶和过氧化氢酶活性受外源腐胺的影响很小,两者的活性变化与PAO活性变化趋势一致.