棉花ms5ms6雄性不育两用系发育过程中POD同工酶分析

为了在生长发育前期简便快速地区分棉花雄性不育A、B两用系的不育株和可育株,采用同工酶分析方法,对POD同工酶进行研究。结果表明,不同器官之间POD同工酶带数及其带的强度存在差异,其中花药的酶带最多、带的强度最高;子房的酶带最少,带的强度最低。同一器官不同发育时期POD同工酶也存在差异,其中在花药发育时期POD同工酶带数及其带的强度差异明显,尤其在单核期和二核期两种不同叶型的两用系A比B分别减少1或2条带。因此,在开花之前根据花药POD同工酶的差异,易于区分A、B两用系中的不育株和可育株。     

Isoenzymes of Japanese-radish Peroxidase

Japanese-radish root contained eighteen isoenzymes of peroxidase distinguishable on polyacrylamide gel electropherograms. The isoenzymes were found to be quite similar to those of horseradish peroxidase, although their quantities were different between two plants. The acidic components were the major isoenzyme in Japanese-radish peroxidase, while the neutral ones were the major one in horseradish. The chromatographic purification of the isoenzymes was performed on CM- and DEAE-Sephadex columns to characterize the components. The components in the preparations purified by the previously reported procedures of Morita et al. were also identified.     

Peroxidase Isoenzymes in Strawberry Roots Infected with Binucleate Rhizoctonia spp. and their Implication in Disease Resistance

Infection of strawberry plants with binucleate Rhizoctonia spp. results in an increase in peroxidase activity and the appearance of new isoforms of the enzyme. In healthy and diseased roots of two different strawberry genotypes seven peroxidase isoenzymes were found. In healthy strawberry cv. Senga Sengana, which was moderately resistant to infection, four isoenzymes (1, 2, 5, and 6) were found. Moreover the activity of these isoenzymes was increased and three new isoenzymes (3, 4, and 7) were found in infected roots. In the strawberry hybrid 3/2/86/88/R, which is very susceptible to infection, only isoenzyme 2 was present in the roots of healthy plants. Following infection, the activity of isoenzyme 2 was increased and five new isoenzymes (1, 4, 5, 6, and 7) were detectable. The results obtained indicate that strawberry resistance to binucleate Rhizoctonia may be correlated with peroxidase isoenzyme profile with particular reference to isoform 3, which is only present in infected roots of the moderately resistant cv., Senga Sengana.     

心里美萝卜营养生长期肉质根过氧化物酶同工酶谱及组化分析

过氧化物酶［peroxidase,POD,EC 1. 11. 1. 7 (X)］为一类多基因家族的同工酶,其聚合形式对其功能具有重要影响．心里美萝卜(Raphanus sativus var.L) 肉质根富含花青素和POD等．本文采用不同的电泳技术、组化定位和活性测定研究了心里美萝卜营养生长期肉质根中POD在不同时期和不同组织中的表达、分布特点及同工酶的组成和性质．结果显示,心里美萝卜营养生长期肉质根的皮和肉中POD活性均先升高后降低,分别在播种后第40 d和60 d出现峰值,30 d时皮中POD活性高于肉,之后则相反,酶活性最高时同工酶数目也最多,不同时期,皮和肉中同工酶种类不同;皮中POD同工酶由5个单体和4个二聚体组成,而肉中由5个二聚体或2个单体、2个二聚体和1个四聚体组成,均以酸性同工酶为主;组化定位显示POD主要分布在肉质根的周皮、形成层、木射线、木质部导管和肉的初生木质部等的细胞壁附近．从上述结果得出结论： 酸性POD同工酶在植物体中以多种形式存在．     

Do S allele-specific peroxidase isoenzymes exist in self-incompatible Nicotiana alata?

Summary In order to test Pandey's hypothesis that peroxidase isoenzymes determine S-gene specificity in Nicotiana alata, peroxidase isoenzymes in styles and pollen from various plants of an inbred- and a cross progeny were compared by means of starch gel electrophoresis and electrofocusing.No relation between the S-genotype and the peroxidase isoenzyme patterns of pollen or of styles could be established. The differences between the isoenzyme patterns of different S-genotypes were ascribed to differences in the genetic background of various plants that had the same S-genotype.     

Activity, isoenzyme composition, thermostability and molecular weight of peroxidase from intact and virus infected tobacco plant leaves]

The enzyme peroxidase was isolated from the leaves of the tobacco plant Xanthi (intact and infected with weakly (XY) and highly (XT) pathogenic strains of potato X-virus) and partially purified. The original extract (the 30,000 g supernatant) was purified by ammonium sulfate at 30--80% of saturation and by gel filtration through Sephadex G-25 and G-100 in 0.05 M tris-HCl buffer, pH 7.4 containing 17% sucrose. Disc electrophoresis revealed that both intact and infected plants contain 10 isoperoxidases. The electrophoregrams of isoenzymes from infected plants with the Rf values of 0.1, 0.48, 0.53 and 0.59 stained with benzidine produced a more intensive colouring as compared to the corresponding isoenzymes from intact plants. The total enzymatic activity for the plants infected with the XY and XT strains made up to 180% and 240% of that for the intact plants, respectively. The molecular weights of the peroxidase isoenzymes were found to be the same and equal to 40,000. Study of the thermostability at 60 degrees C and pH 7.0 showed that after 90 min the enzyme activity was 12.4% and 5.1% of the original one in intact and infected plants, respectively. The data obtained suggest that the activity, thermostability and synthesis of some peroxidase isoenzymes in tobacco plant leaves are affected by viral infection.     

Myrosinase in Brassicaceae (Cruciferae): II. MYROSINASE ACTIVITY IN DIFFERENT ORGANS OF SINAPIS ALBA L.

Seeds of Sinapis alba L. were germinated in darkness for 3 dand a part of the etiolated seedlings were transferred to long-dayconditions for 6 weeks. Myrosinase solutions were prepared forcotyledons, hypocotyls, primary roots, leaves, stems, inflorescences,and seeds, and used to hydrolyse sinigrin. Glucose, one of the cleavage products, was determined by fourdifferent spectrophotometric methods, and their usefulness forcalculations of myrosinase activity in crude plant extractsis considered. Specific activity was calculated in relations to protein, andit was found to be about 30% higher in seedlings and also higherin seeds than in adult plants. Of the organs, those with thehighest activity were the hypocotyls and the stems. The differentparts of the plant contained different numbers of isoenzymes,as shown by polyacrylamide gel electrophoresis. Seedlings yieldedtwo, and adult plants four or five isoenzymes.     

Plant NAD(H)-Glutamate Dehydrogenase Consists of Two Subunit Polypeptides and Their Participation in the Seven Isoenzymes Occurs in an Ordered Ratio

The structure and function of NAD(H)-glutamate dehydrogenase in plants was studied by using grapevine (Vitis vinifera L. cv Sultanina) callus grown under different nitrogen sources. The enzyme consists of two subunit-polypeptides, α and β, with similar antigenic properties but with different molecular mass and charge. The two polypeptides have molecular masses of 43.0 and 42.5 kilodaltons, respectively. The holoenzyme is hexameric and is resolved into seven isoenzymes by native gel electrophoresis. Two-dimensional native/SDS-PAGE revealed that the 1 and 7 isoenzymes are homohexamers and the isoenzymes 2 through 6 are hybrids of the two polypeptides following an ordered ratio. The total quantity of α- and β-polypeptides and the isoenzymic pattern was altered by the exogenous nitrogen source. The sample derived from callus grown on nitrate or glutamic acid contained a slightly greater amount of β-polypeptide and of the more cathodal isoenzymes, whereas α-polypeptide and the more anodal isoenzymes predominated in callus grown in the presence of either ammonium or glutamine. The anabolic reaction was correlated with the α- and the catabolic reaction with the β-polypeptide; this could suggest that each isoenzyme exhibits anabolic and catabolic function of different magnitude. The isoenzymic patterns did not obey the expected binomial distribution proportions.     

Immunochemical comparison of glutamine synthetases from some solanaceae plants

The presence of different glutamine synthetase isoenzymes in different Solanaceae plants and their relative antigenicities against antiglutamine synthetase from tomato leaf serum were studied. All the plants tested showed one glutamine synthetase isoenzyme except for Mandragora autumnalis, which showed two, after discontinuous polyacrylamide gel electrophoresis and specific in situ assay. Antigenicities were compared by the double immunodiffusion technique. The Nicotiana glauca enzyme showed equal reactivity to that of Lycopersicon esculentum, but its antigenicity was higher than Withania frutescens, Datura stramonium, and Hyoscyamus niger. The study of relative antigenicities permitted differentiation of the glutamine synthetase enzymes from uncultivated species of Solanaceae.     

Cd2+胁迫对小麦幼苗生长及呼吸作用的影响

以小麦品种郑州9023为材料,研究了不同浓度Cd2 胁迫对小麦幼苗生长及呼吸作用的影响.结果显示:(1)随Cd2 胁迫浓度的升高,小麦幼苗根和芽的呼吸速率及琥珀酸脱氢酶(SDH)活性均呈先上升后下降的趋势.(2)Cd2 胁迫对小麦幼苗根中细胞色素氧化酶(COD)、苹果酸脱氢酶(MDH)、异柠檬酸脱氢酶(IDH)同工酶表达的影响较小,都呈低浓度诱导、高浓度抑制的效应,且Cd2 处理诱导了根中新的MDH、IDH同工酶带的表达;而不同浓度Cd2 对小麦幼苗芽中COD、MDH、IDH同工酶的表达影响较小.(3)随Cd2 胁迫浓度的增加,芽长、根长、芽干重、根干重均呈持续下降的趋势,且对根的抑制作用明显大于对芽.研究表明,Cd2 胁迫可以改变小麦幼苗根和芽中SDH、COD、MDHI、DH等呼吸作用关键酶的活性或同工酶表达,从而影响其呼吸作用,最终抑制了幼苗的生长.     

山楂叶螨危害对海棠叶片POD的影响

以人工接种的方法研究了山楂叶螨(Tetranychus viennensis Zacher)危害初期(9d内)对海棠(Malus zumi)叶片过氧化物酶(POD)的影响。结果表明,随危害时间延长和虫口密度增大,POD活性迅速升高,30头／叶危害9d和60头／叶危害6d时均约达对照的190％;PAGE分析结果显示叶螨危害使海棠叶片产生2种新的POD同工酶,受害植株的未接虫叶片的POD活性同时升高,并出现与接虫叶片相同的PAGE谱带,揭示叶螨危害对海棠叶片POD有系统诱导性。POD的变化可能是受害植株对叶螨危害的一种应激反应。     

Metabolism of the human serum beta-hexosaminidase isoenzymes in the rat: an in vivo and in vitro study

Plasma clearance of purified human serum beta-hexosaminidase isoenzymes was studied in the rat. The serum beta-hexosaminidase isoenzymes (A, B and P) showed a slow clearance from circulation compared to their tissue counterparts. After desialylation, the clearance rate of all serum isoenzymes was markedly enhanced. The uptake of native as well as desialylated serum beta-hexosaminidase isoenzymes was studied in rat liver nonparenchymal cells and hepatocytes. No detectable uptake of any native serum isoenzyme was noticed in either cell type. However, when these isoenzymes were desialylated by neuraminidase treatment, isoenzymes A and B were taken up by the nonparenchymal cells. No uptake was observed for the P form. None of the desialylated serum forms was taken up by hepatocytes.     

Soluble peroxidase gradients in lupin hypocotyls and the control of the level of polarly transported indole-3yl-acetic acid

The distribution of basic soluble isoperoxidases along the growth gradient of lupin hypocotyl was studied in order to establish the role of these isoenzymes in controlling polarly transported indole-3yl-acetic acid (IAA) levels. The observation that the levels of basic isoperoxidases, which diminish from the young (vascular differentiating) to the older (vascular differentiated) tissues, are related with previously reported IAA oxidation rates in decapitated plants, suggests that these isoenzymes can play a role in the oxidation of IAA during polar transport. The fact that the level of basic isoperoxidases is controlled by IAA in hypocotyl sections harvested from different growth zones is in accordance with the previously described adaptative activation of basic isoperoxidases to IAA content. This adaptative activation of basic isoperoxidases might constitute the basic characteristic of a system of subcellular oscillators, coupled at the cellular level, necessary to generate the supracellular auxinwave associated with auxin transport.     

Soluble peroxidase gradients in lupin hypocotyls and the control of the level of polarly transported indole-3yl-acetic acid

The distribution of basic soluble isoperoxidases along the growth gradient of lupin hypocotyl was studied in order to establish the role of these isoenzymes in controlling polarly transported indole-3yl-acetic acid (IAA) levels. The observation that the levels of basic isoperoxidases, which diminish from the young (vascular differentiating) to the older (vascular differentiated) tissues, are related with previously reported IAA oxidation rates in decapitated plants, suggests that these isoenzymes can play a role in the oxidation of IAA during polar transport. The fact that the level of basic isoperoxidases is controlled by IAA in hypocotyl sections harvested from different growth zones is in accordance with the previously described adaptative activation of basic isoperoxidases to IAA content. This adaptative activation of basic isoperoxidases might constitute the basic characteristic of a system of subcellular oscillators, coupled at the cellular level, necessary to generate the supracellular auxinwave associated with auxin transport.     

Phytochelatin synthesis and responses of antioxidants during arsenic stress in <Emphasis Type="Italic">Nasturtium officinale</Emphasis>

The induction of thiols including glutathione and phytochelatins as well as the activities of antioxidant enzymes namely superoxide dismutase isoenzymes and those enzymes involved in the ascorbate-glutathione cycle in watercress plants under arsenic stress were investigated. Arsenic concentrations and tissue type-dependent response to arsenic were assessed. Plant was capable of accumulating large amounts of arsenic in the shoots. Superoxide dismutase isoenzymes activity and phytochelatin level was higher in shoots than in roots. In roots, ascorbate levels increased significantly, while no relationship was found between ascorbate contents and arsenic tolerance in shoots. Treatment with arsenic resulted in a remarkable increase in glutathione content of roots at all of the arsenic concentrations, while in shoots, glutathione content increased by lower levels of arsenic. Differences were noted in both roots and shoots for enzymes involved in the ascorbate-glutathione cycle. These results suggest that, the strategy of tolerance to arsenic toxicity in roots of watercress plants is different from that of shoots.     

Purification and properties of human kidney-cortex hexosaminidases A and B.

Hexosaminidases (EC 3.2.1.30) A and B from human kidney cortex were purified to homogeneity by using concanavalin A affinity chromatography, ion-exchange chromatography and gel filtration. The yield of homogeneous isoenzymes improved approx. 20-fold, giving preparations of hexosaminidases A and B with specific activities of about 200 and 325 units/mg of protein respectively. The kinetic and structural properties of kidney hexosaminidase isoenzymes were studied and compared with the hexosaminidase isoenzymes from human placenta. The amino acid composition of hexosaminidase A was significantly different from that of hexosaminidase B. In the event of success in developing enzyme-replacement therapy for Tay-Sachs and Sandhoff's diseases, this modified procedure can furnish larger amounts of homogeneous isoenzymes.     

Mechanism of peroxidase isoenzyme induction in pollinated Nicotiana alata styles

Summary A comparative study on the induction of peroxidase isoenzymes, specifically number 10 (P-10) in Nicotiana alata styles revealed significant differences between the various plants of an inbred progeny. In some plants the ageing-induced increase in P-10 activity was very low, whereas in some others, it was relatively high. Pollination accelerated this increase, independent of the pollen genotype. Fertilization was followed by a considerable increase in the activity of several peroxidase isoenzymes, including P-10 in all the plants.Two plants that differed greatly with regard to P-10 induction were used in additional experiments in order to ascertain the mechanism involved in the induction of P-10. The increase in P-10 activity due to pollination or fertilization can partly be explained on the basis of auxin and auxin-induced ethylene activity. The differences in P-10 induction between various plants of the inbred progeny were probably due to differences in their sensitivity to ethylene.     

Bioelicitors Induce Association of Defence Enzymes with Cell Walls of Lycopersicum esculentum

Cell walls of plants are complex structures impregnated with various proteins having wide array of functions. In this study, twenty‐eight proteins isolated from tomato cell walls were subjected to MALDI‐TOF MS followed by mass peak analysis using ORIGIN 6 software. The mass peaks subjected to MASCOT and ProFound databases for peptide mass fingerprinting led to the identification of 9 protein domains. These proteins were further classified according to their functions. Fruit extracts of A. indica could elicit induction, localization and functioning of peroxidase (POX) and polyphenol oxidase (PPO) and their isoenzymes in cell walls of Lycopersicum esculentum (tomato) against Pseudomonas syringae pv. tomato. The results revealed the possible involvement of cell wall‐bound proteins in defence of plants against the invading pathogens. A number of novel isoenzymes of both POX and PPO were found to be located in the cell walls of the plants treated with neem extract. Neem extract can induce accumulation and binding of isoenzymes to cell walls. These isoenzymes could possibly protect host plants against the invading pathogens.     

Detection of Somaclonal Variations in Tissue Culture-Derived Date Palm Plants Using Isoenzyme Analysis and RAPD Fingerprints

Isoenzyme analysis and activities of peroxidase (PER), polyphenol oxidase (POD) and glutamate oxaloacetate (GOT) and randomly amplified polymorphic DNA (RAPD) fingerprints were used to analyze somaclonal variations in tissue culture-derived date palm plants. The frequency of somaclonal variations was found to be age dependent. Similar isoenzyme patterns for PER and GOT were detected in all analyzed plants. However, variations in activities of the three enzymes studied and in POD isoenzymes were detected. RAPD analysis showed genetic variations in approximately 4 % of the analyzed plants (70 regenerants). The genetic variations were only detected in 6- and 12-months-old cultures. It was observed also that all morphologically abnormal shoots showed genetic variations at the molecular level.