Characterization of isoperoxidase activity in Arachis hypogaea cultivars

Isoperoxidases (both anodic and cathodic) from individual seeds of several peanut cultivars (Arachis hypogaea), after ammonium sulfate precipitation of the major storage protein, arachin, from the extracts were characterized by polyacrylamide gel electrophoresis with respect to seed development, maturity and germination and the geographic areas where grown. All cultivars had a major cathodic isozyme near the origin of the gels. Much anodic intra- and inter-varietal isozyme polymorphism was observed in mature seeds collected from different geographic areas. These polymorphic isozymes were consistently present during the earlier stages of seed development (the activities decreased quantitatively and became variable during the later stages of maturation), and most were observed in each peanut upon germination, the latter showing quantitative increases in activity.     

Developmental expression and biochemical characterization Nassarius obsoleta.

Electrophoretic analysis of catalase isozyme patterns of Nassarius obsoleta indicates that these isozymes are products of two separate loci. Five aminopeptidase isozymes have also been detected in N. obsoleta and the data suggest that at least two loci encode these aminopetidase isozymes. Experiments designed to test for the interconvertibility of the isozymes indicated that the two catalase isozymes and that two of the five aminopeptidase isozymes tested were not conformational artifacts but distinct genetic products. No change in catalase isozyme expression, but considerable quantitative variation in catalase specific activity was noted during early developmental stages of N. obsoleta. Both qualitative and quantitative changes were noted in aminopeptidase expression during early developmental stages. This paper also describes several physicochemical parameters for each of the two enzymes under study.     

Seed esterases,leucine aminopeptidases and catalases of species of the genus Gossypium

Summary Polyacrylamide and starch gel electrophoresis were used to analyze the isozyme makeup of three enzyme systems (esterases, leucine aminopeptidases and catalases) from the dormant seeds of twenty-nine species within the genus Gossypium.Isozyme variation was observed for all three enzymes between the species of the different genome groups. The within species polymorphism noted for the esterases was not observed for the leucine aminopeptidase and catalase patterns. In general, only minor qualitative banding pattern differences distinguished the A and B genome species, whereas, band variations were greatest between the more distantly related species in the C, D and E genomes. Gossypium longicalyx (F genome) showed an overall banding pattern unique to itself. The species of the genomes (C, D, E and F) removed from the postulated area of genetic origin (Southern Africa) also exhibited greater isozyme variability than that of the wild species of the A and B genomes, both located in Southern Africa.Synthetic mixtures of seed extracts from parent species of recently formed synthetic allopolyploids produced additive isozyme patterns for esterase, leucine aminopeptidase and catalase that were closely comparable to the zymograms produced by their hybrids. In contrast all three enzyme systems showed significant qualitative isozyme variations between the three natural allotetraploids, G. tomentosum, G. barbadense and G. hirsutum when compared to the zymograms of the synthetic mixtures of their alleged parental forms.This paper is part of a dissertation by the first author for the degree of Doctor of Philosophy in Genetics. Journal paper 1763 of the Arizona Agricultural Experiment Station.National Research Council Postdoctoral Research Associate.     

Study on Heat Sensitivity of Esterase Isozymes in Maise Inbreds

Crude extracts of 44 maize inbred seeds were treated under different temperatures, i. e. 50, 55, 60, 65 and 70℃, in order to determine their heat sensitivity of esterase isozymes. The results showed large discrepancy of heat sensitivity of anodic esterase isozymes between different or same relative mobility bands of isozyme, but similar inactivating temperature (65℃) of the cathodic ones for different inbreds. The heat sensitivity of isozymes for inbreds may be adopted in the identification of variation and purity of inbred and hybrid seeds of maize.     

Plant Leaf and Stem Proteins. II. Isozymes and Environmental Cabbage

The activity of 10 enzymes separated by acrylamide disc gel electrophoresis of leaf and stem extracts from Dianthus grown under summer and winter conditions was studied. While banding was constant and highly reproducible under each environment, differences between the 3 cultivars and between the tissues were evident. No significant differences in the isozyme patterns of glutamate dehydrogenase, 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, malate dehydrogenase, and catalase were observed between the 2 environments. Loss of activity was observed under winter conditions with amylase and lactate dehydrogenase and loss of certain isozymic components was evident with acid phosphatase and esterase. Prominent changes were observed in peroxidase isozymes, the hardy cultivars developing additional isozymic components under winter conditions. Only minor changes in the total protein banding were seen. The enzymes showed considerable stability in those tissues killed by the freezing conditions.     

Genetische Untersuchungen bei Picea abies mit Hilfe der Isoenzym-Identifizierung

Summary Using techniques of starch gel zone electrophoresis, a considerable variation among esterase and leucine aminopeptidase isoenzyme patterns was found in the endosperm of dormant seeds of Norway spruce (Picea abies). Since the so-called primary endosperm of conifers is only a further developmental stage of the haploid female gametophyte, simple Mendelian segregations can be determined in seeds of individual open-pollinated trees. It was therefore possible to identify some esterase and leucine aminopeptidase loci only with regard to phenotypic frequency distributions without difficult crossing procedures.

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Angenommen durch G. Melchers     

新疆干旱区植物藜的种子异型性及其萌发机理

新疆干旱区分布的植物藜(Chenopodium album)的种子有黑色和褐色两种类型。对藜的异型性种子从形态结构、不同环境因素及激素或化学物质对萌发的影响以及同工酶谱等方面进行了研究,并对其萌发及适应异质环境的机理进行了讨论。结果表明:(1)藜的异型性种子在形态结构、萌发休眠特性等方面都存在明显差异:黑色种子种皮厚且硬,休眠,萌发慢,萌发率低;褐色种子种皮薄而软,不休眠,萌发快且萌发率高;(2)黑色种子的休眠可通过切除胚根外缘种皮得以完全解除;(3)赤霉素、乙烯利对黑色种子的萌发无明显促进作用;KNO3可较显著促进黑色种子的萌发;协同使用乙烯利和KNO3时,可显著提高黑种子萌发率,完全打破休眠;(4)黑色种子和褐色种子的酯酶、过氧化物酶及过氧化氢酶同工酶谱带存在差异;(5)黑色种子的萌发需要光照,而褐色种子则对光不敏感;低温贮藏对二者的萌发均无显著影响,尽管黑色种子的萌发率有波动。研究结果初步显示黑色种子的休眠是内源(胚)和外源(种皮)因素共同所致。藜的种子异型性及其萌发机理的形成是其对新疆干旱区异质化环境的高度适应。     

Acid phosphatase and leucine aminopeptidase isozymes as biochemical markers of homogeneity in oil seed rape androgenetic lines

Extracts of cotyledons of Brassica napus plants (seed progenies of doubled haploid plants) were separated by electrophoresis on polyacrylamide gels and stained for acid phosphatase (ACP-E.C. 3.1.3.2.) and leucine aminopeptidase (LAP-E.C. 3.4.11.1.) enzymes to investigate the possibility of utilising isozymes as markers of homogeneity (purity) of plant populations. One zone of activity for acid phosphatase and two zones of activity for leucine aminopeptidase were identified on gels, some variation in isozyme patterns occurred in several androgenetic lines. This method is appropriate and consistent for testing the homogeneity of breeding lines-progenies of double haploid (D.H.) plants.     

Acid phosphatase and leucine aminopeptidase isozymes as biochemical markers of homogeneity in oil seed rape androgenetic lines

Extracts of cotyledons of Brassica napus plants (seed progenies of doubled haploid plants) were separated by electrophoresis on polyacrylamide gels and stained for acid phosphatase (ACP-E.C. 3.1.3.2.) and leucine aminopeptidase (LAP-E.C. 3.4.11.1.) enzymes to investigate the possibility of utilising isozymes as markers of homogeneity (purity) of plant populations. One zone of activity for acid phosphatase and two zones of activity for leucine aminopeptidase were identified on gels, some variation in isozyme patterns occurred in several androgenetic lines. This method is appropriate and consistent for testing the homogeneity of breeding lines-progenies of double haploid (D.H.) plants.     

盐胁迫对花生种子际细菌菌群结构的调控

盐胁迫影响种子萌发和植株形态建成,提高盐胁迫下花生种子萌发速率和成苗健苗率是盐碱地花生高产高效栽培的重要环节之一,花生种子际土壤细菌菌群结构与种子萌发关系密切。为揭示盐胁迫对花生种子际微生物菌群结构的影响,以耐盐花生品种（花育25号,HY25）和盐敏感花生品种（花育20号,HY20）为试验材料,采用盆栽实验和高通量测序技术,研究不同耐盐性品种种子萌发吸胀吸水阶段种子际细菌菌群结构的变化。结果表明,种子际土壤细菌群落以变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）、放线菌门（Actinobacteria）、拟杆菌门（Bacteroidetes）及芽单胞菌门（Gemmatimonadetes）等为优势菌门,盐胁迫处理可以不同程度的提高厚壁菌门和放线菌门的相对丰度。在属水平上,盐胁迫可以增加有益菌芽胞杆菌属（Bacillus）的相对丰度,增强盐胁迫下种子存活能力,提高萌发率。细菌功能预测结果显示,信号转导机制、免疫系统和防御机制等相关功能在盐胁迫处理后明显增强,可能是促进花生萌发并增强花生胁迫应答的重要原因之一。种子际优势菌群的鉴定及机理分析可为通过改良种子际土壤微生物环境,提高花生耐盐性和出苗健苗率提供重要的借鉴意义,同时为开发利用盐碱地提供参考。     

湖北大麦品种酯酶同工酶酶谱与地区分布关系的研究

对产于湖北的１０５份大麦品种,采用聚丙烯酰胺凝胶电泳进行了酯酶同工酶酶谱分析,共得到８种酶谱类型。８种酶谱聚为４类,其中具酶谱类型Ｉ、Ⅱ、Ⅲ和Ⅷ的品种约占供试材料的９４％,构成湖北大麦酯酶同工酶酶谱的主要类型。对这些品种来源地的分析表明,具有相同或相近酶谱的品种分布在一些相对集中的地区,这些地区的气候条件较相似。对湖北大麦酯酶同工酶酶谱类型与地区分布和气温关系的关系,以及酯酶在大麦引种育种生产实践     

Studies on the Molecular Weights of Esterase and Peroxidase Isozymes of Maize

The molecular weights of esterase and peroxidase isozymes of maize seedlings were directly determined by improved polyacrylamide gradient gel electrophoresis. The different isozyme bands developed in polyacrylamide slab gel electrophoresis (uniform gel) were identified in polyacrylamide gradient gel electrophoresis by means of isozyme variants. The molecular weights of esterase isozymes E1, E2, E3F, E3S, a, b, c, named according to isozyme patterns in uniform gel, are <20000, 35200, 33000, 38500, 29900, 28500, 34000 doltons respectively. The molecular weights of peroxidase isozymes PX4F and PX4S are 131000 and 149000 doltons respectively. According to the band location in uniform gel and in gradient gel, some biochemical properties of the isozyme bands and relationships between the isozyme bands were analyzed. The possible errors in the determination of smaller molecular weight isozymes are discussed.     

Isoenzyme Polymorphism in Flowering Plants I. Diffusion of Enzymes Out of Intact Pollen Grains

Esterases, leucine aminopeptidases, catalases, amylases and acid phosphatases diffuse out of intact and ungerminated pollen grains of Oenothera organensis, whether suspended in 1 % sodium chloride or in pollen medium. A total of 15 esterase isozymes are recorded; 5 of them appear within 5 minutes, 8 within 30 minutes, 9 within 2 hours, and 13 within 19 hours. Pollen grains suspended for 19 hours gave much stronger isozyme bands than macerated pollen grains. However, one esterase hand was consistently missing from the 19 hour suspensions, although present in all others. It is suggested as a working hypothesis that the early growth of pollen tubes and possibly even germination of pollen grains may be influenced by the metabolic products of pistillate tissues caused by the diffused pollen enzymes, and that inactivation of these enzymes by stigmatic or stylar components could lead to incompatibility reactions.     

玉米酯酶同工酶和过氧化物酶同工酶的分子量研究

本试验利用聚丙烯酰胺凝胶梯度电泳分步染色法直接对玉米苗期酯酶同工酶和过氧化物酶同工酶各酶带的分子量进行了比较测定。酯酶同工酶 E_1、E_2、E_3~F、E_3~S、a、b、c 各酶带的分子量分别为<20000,35200、33000、38500、29900、28500、34000道尔顿过氧化物酶同工酶 PX_4~F和 PX_4~S酶带的分子量分别为131000和149000道尔顿。根据酶带在均匀胶和梯度胶中的位置变化对各酶带的生化性质作了初步分析,发现 E_3~F和 E_3~S、PX_4~F 和 PX_4~S 在迁移率上的差异主要是分子量的差异。本文为同工酶的分子量测定提供了一个简便的方法。     

苜蓿组织培养中球形胚发生时特异蛋白质和同工酶分析

试验在苜蓿组织培养中,对球形胚形成过程中特异蛋白质表达的模式、过氧化物酶及酯酶同工酶酶谱变化进行研究,结果表明：苜蓿组织培养中从胚性愈伤组织到球形胚发育的进程中,顺序消失和出现了11种中小分子量多肽;过氧化物酶同工酶酶谱发生了显著的变化;酯酶同工酶酶谱变化不大,但其总活力对于维持体细胞胚胎发生是必须的。     

COMPARATIVE ELECTROPHORESIS AND ISOZYME ANALYSIS OF SEED PROTEINS FROM CULTIVATED RACES OF SORGHUM

Comparative disc electrophoresis of acidic proteins, basic proteins, and isozymes of esterase, MDH, and peroxidase were performed with aqueous extracts of seeds from seven cultivars belonging to five races of Sorghum bicolor ssp. bicolor: bicolor, caudatum, durra, guinea, and kafir. Two disc electrophoretic systems were employed. Acidic proteins were electrophoresed in an anionic system (tris-glycine buffer, pH 8.3). Basic proteins were electrophoresed in a cationic system (β-alanin-acetate buffer, pH 4.5). Soluble proteins were stained with Coomassie brilliant blue. Isozyme activity was detected by using specific enzyme stains and substrates. Each cultivar yielded reproducible, characteristic patterns of distinct acidic and basic proteins. Cultivars belonging to the same race produced identical protein and isozyme patterns. The degree of electrophoretic similarity among races was estimated by calculating similarity index values for each of the 10 possible pairs of races. Bicolor, caudatum, durra, and guinea produced very similar acidic and basic protein patterns and esterase, MDH, and peroxidase isozyme patterns. Differences, however, were observed among all races. All of kafir patterns were significantly different from the patterns of other races. Comparative electrophoresis may provide a new source of taxonomic characters for investigating phenetic and phylogenetic relationships in Sorghum.     

中国苋属植物酯酶同工酶研究

用聚丙烯酰胺凝胶盘状电泳技术对中国苋属植物１３种的酯酶同工酶谱进行了研究。结果表明：苋属植物酯酶同工酶有２条属的标志带在生化水平上认为是一个自然的分类群;     

Isozyme Profile During Somatic Embryogenesis and In Vitro Flowering of Bambusa vulgaris

Peroxidase and esterase isozymes were investigated during plant regeneration via somatic embryogenesis in Bambusa vulgaris, The transition of non-embryogenic calli to embryogenic calli, somatic embryo development, germination and subsequent flowering of somatic embryo derived shoots were associated with selective expression or repression of isoforms of peroxidase and esterase. Non-embryogenic callus showed six peroxidase and four esterase bands. During somatic embryogenesis and germination of somatic embryos, some bands were suppressed and new isoforms of peroxidase and esterase appeared. During flowering, in addition to four peroxidase bands, a new unique esterase band ‘a’ appeared. Each developmental stage was thus associated with a definite isozyme profile.     

赖草属七个种同工酶研究

用聚丙酰胺凝胶电泳法对赖草属７个种的幼根、幼叶进行了酯酶、过氧化物酶同工酶分析,结果表明：无论从相同器官进行不同的同工酶分析,还是从不同器官进行相同的同工酶分析,这７个种的酶谱在各带区存在相似酶带,但更多的是相异酶带。从酯酶、过氧化物酶这两种酶分析结果来看,酯酶比过氧化物酶分离效果好些;从幼根和幼叶这两个器官的酶谱来看,幼根比幼叶酶带多些,分离效果也好些。同时也表明这７个种的酶谱变化与染色体倍性变化无关     

Esterase activity and isozymes during the ontogeny of stamens of male fertile lycopersicon esculentum mill., A male sterile stamenless-2 mutant and the low temperature-reverted mutant

A comparative study of the activity and isozymes of esterase during the ontogeny of stamens of normal male fertile (MF) Lycopersicon esculentum, a male sterile stamenless-2 (sl-2/sl-2) mutant (MS), and the low temperature reverted mutant was conducted. In MF and MS stamens, a major isozyme associated with tapetum development was present at early stages whereas another isozyme related to pollen formation was observed at later stages of development. There was a difference, however, in the timing of the appearance of these isozymes in MF and MS stamens. Also, the number and intensity of most bands, and the overall and specific activity of esterase was higher in MF than in MS stamens. The isozyme pattern, the number and intensity of bands and the overall activity of esterase was comparable in MF and MS-reverted stamens from low temperature grown plants. Male fertile stamens from low temperature-grown plants contained lower esterase activity than those grown in the greenhouse. The results are discussed with respect to the mechanism of male sterility in the sl-2/sl-2 mutant of tomato.