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.     

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。     

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.     

菊花不同生长阶段不同器官POD和EST同工酶比较

采用过氧化物酶(POD)、酯酶(EST)2个酶系统的12个同工酶位点,分析了4个菊花品种营养生长和生殖生长阶段不同器官(嫩叶、老叶、嫩茎、木质化茎)的同工酶变化.结果表明:(1)4个品种共有16种POD酶带,15种EST酶带;(2)菊花的POD和EST具有组织特异性和阶段特异性,其中以嫩叶的酶带最多,其次为老叶,再次为嫩茎,而木质化茎的酶带最少;(3)与生殖生长阶段相比,营养生长阶段的POD酶带更清晰,更整齐,分离更好,但生殖生长阶段的EST同工酶比营养生长阶段的更清晰;(4)营养生长阶段的嫩叶最适合用于菊花POD同工酶分析,而EST同工酶研究宜取生殖生长时期的嫩叶.     

Identification of Flax and Linseed Cultivars by Isozyme Markers

A set of 28 fibre flax and linseed cultivars differing in plant morphology and technological parameters were analysed by isozyme markers in five ontogenetic phases. Relatively high isozyme polymorphism was observed using polyacrylamide gel electrophoresis. Altogether 18 isozyme systems produced 145 different bands; 66 of them (45.52 %) have been found to be polymorphic. The highest level of polymorphism was found in acid phosphatase and esterase, polymorphism was detected in aconitase, diaphorase, glutamate dehydrogenase, peroxidase and superoxide dismutase as well. The highest number of unique isozymic spectra (cultivar × enzyme × ontogenetic phase) was detected in the phase of shoot with removed cotyledons. Electrophoretic analysis of all polymorphic isozymes enabled to distinguish 20 cultivars (71 %) in the screened cultivar set.     

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.     

Isozyme modifications during morphogenesis of callus from barley and wheat

Callus cultures fron non-organogenic, young and one-year old, and morphogenic calli were used to assess the value of isozymes analysis for the prediction of morphogenic capacity by studying esterase, peroxidase and acid phosphatase. Basic isozyme patterns of each enzyme for the callus were retained in all the callus stages and in the callus which has differentiated into shoots. With the development of shoot and/or root some conspicuous isozymes appeared for esterase and acid phosphatase and some disappeared for peroxidase. As the isozyme changes became apparent only after shoot or root initiation these enzymes could not be used as markers to distinguish between morphogenic and non-morphogenic calli.     

Changes of peroxidase,esterase isozyme activities and some cell inclusions in regenerated vascular tissues after girdling inBroussonetia papyrifera (L.) Vent

The isozyme zymogram of peroxidase and esterase, and some cell inclusion contents were changed with the differentiation of regenerated vascular stem tissues after girdling inBroussnetia papyrifera (L.). Vent. Presence or disappearance of some peroxidase and esterase isozyme bands was related to wounding. Some isoperoxidase bands disappeared at the time of vascular tissue formation, but some esterase isozyme bands appeared in phloem or cambial regions as sieve-like elements or mature xylem were formed. The inclusion grains progressively disappeared with the formation of callus and initiation of vascular meristems. The cell inclusions reappeared during the formation of regenrated vascular tissues. Histochemical study indicated that the inclusion grains could be a complex compound of a protein mass encircling polysaccharide in the center with a proteinous nucleus.     

Differential expression of esterase and MDH isozymes during in vitro culture in maize (Zea mays L.)

Isozyme patterns of esterase and malate dehydrogenase were analyzed at different stages of in vitro culture of immature embryos and glumes of Zea mays L. viz. explant, callus formation, root formation and shoot formation. Significant changes in isoenzyme patterns of esterase and MDH were observed besides the appearance of specific and new isozymes. Specific fast migrating isozymes were noted in differentiating calli of embryo and glume calli which were absent at other stages suggesting a possible association of these isozyme patterns with in vitro differentiation.     

Isozyme modifications and plant regeneration through somatic embryogenesis in sweet potato (Ipomoea batatas (L.) Lam.)

Summary The potential of somatic embryogenesis was evaluated for 10 cultivars of sweet potato through extensive embryogenic response and isozyme analysis. Embryogenic callus was induced by incubating lateral buds on Murashige and Skoog medium containing 10 M 2,4-dichlorophenoxyacetic acid for 6–8 weeks. The frequency of embryogenic response was low, and varied with genotypes, ranging from 0 to 17%. Embryo to plantlet formation could be enhanced by the use of the combination of 2,4-dichlorophenoxyacetic acid with kinetin, both used at 0.01 M. Embryogenic callus with its potential of plantlet formation has constantly been maintained for over two years. However, after several subcultures, 0.5 to 12% of embryogenic callus reverted irreversibly into friable fast-growing non-embryogenic callus whose ability to regenerate shoots was then definitively lost. The isozymes of esterase, peroxidase, glutamate oxaloacetate transaminase and acid phosphatase investigated in this study were found appropriate to distinguish compact embryogenic from friable non-embryogenic callus in sweet potato. In fact, the callus reversion was associated with a loss of bands or a decline in isozyme activity. On the contrary, very small changes in isozyme activity or no specific changes at all were observed during the differentiation of embryogenic callus into globular embryos.Abbreviations Acp acid phosphatase - BAP 6-benzylaminopurine - cv cultivar - df degree of freedom - 2,4-D 2,4-dichlorophenoxyacetic acid - Est esterase - Got glutamate oxaloacetate transaminase - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - Prx peroxidase - Tris tris(hydroxymethyl)aminomethane     

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     

罗汉果雌雄株同工酶性别鉴定研究

采用电泳技术结合同工酶染色,分析了罗汉果雌雄株叶片的过氧化物酶同工酶、酯酶同工酶、超氧化物歧化酶同工酶、多酚氧化酶同工酶和过氧化氢酶同工酶。结果表明:罗汉果雌雄叶片在同工酶谱上,存在着与性别性状相关的酶带;雌雄间的差异酶带在每一种同工酶中均有一条以上,可作为罗汉果雌雄株间的性别鉴定。此外,还比较了高产、低产、不结果雌株之间同工酶的酶带和活性差别。     

Distorted segregation of the esterase isozyme genotypes in barley (Hordeum vulgare L.)

Distorted segregation of the esterase isozyme genotypes was observed in F2 population which was produced from a cross combination between 'Ko A' and 'Mokusekko 3' of barley varieties. After examining the segregation of esterase isozyme genotypes in B1F1 hybrids derived from the reciprocal backcrosses, it was made clear that the distorted segregation was caused by certation between pollens of different genotypes, independent of the female genotypes. Furthermore, the certation was controlled by a newly designated gene, Ga2, at the locus which was linked with the multiocus, Est1, Est2 and Est4, for esterase isozymes at the long arm of chromosome 3. The distorted segregation ratios did not significantly vary among F2 populations derived from F1 hybrids of the same combination which were grown in different years. Mechanically mixed pollens of the parents, however, could not induce the certation.     

Defense response of resistant and susceptible genotypes of castor (Ricinus communis L.) to wilt disease

The biochemical basis of resistance in castor (Ricinus communis L.) to Fusarium wilt, caused by the pathogen Fusarium oxysporum f. sp. ricini, was investigated. Induction of plant defence against pathogen attack is regulated by a complex network of different signals. Thus changes in various biochemical defenses including antioxidant enzymes, phenolic compounds and pathogenesis related (PR) proteins were investigated in the roots of resistant and susceptible genotypes of castor at 0, 24, 48 and 72 h.a.i. Infection by F. oxysporum significantly increased the superoxide dismutase (SOD) and peroxidase (POX) activities in the roots of susceptible genotypes, while the catalase (CAT) activities were appreciably higher in the roots of resistant genotypes at different stages. Constitutive levels of ascorbate peroxidase (APX) and polyphenol oxidase (PPO) were higher in the resistant genotypes. Also, the activities of phenylalanine ammonia lyase (PAL) and β 1, 3 glucanase significantly increased in the roots of the resistant genotypes after infections. The rate of increment of thiobarbituric acid reactive substances (TBARS) was higher in resistant genotypes after infection. Analysis of isozyme banding pattern of SOD, POX, PPO and esterase on native PAGE electrophoresis revealed that interaction between plant and fungi invoked various isozymes at 48 h of infection. SOD 3 was observed only in resistant genotypes at 24 h.a.i. except Geeta. Similarly induction of POX 5 was observed only in resistant genotypes at 48 h of infection, though the intensity of POX 5 was very less.     

以靛酚乙酸酯为底物的酯酶同工酶显色方法的研究

建立一种以靛酚乙酸酯为底物的酯酶同工酶的显色新方法。酯酶样品的聚丙烯酰胺凝胶电泳（PAGE）凝胶用磷酸缓冲液漂洗约10min后,浸入含有0.002%靛酚乙酸酯的溶液显色5～10min,可显出清晰的蓝色酯酶带。先将酯酶凝胶板浸于有机磷农药溶液中,然后再用靛酚乙酸酯显色液显色,比较同工酶谱,从同工酶带由深蓝色变为浅蓝色的颜色变化,可以看出对有机磷农药敏感的同工酶所受到的抑制程度。     

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.     

Ethylene Production,Peroxidase Activity and the Change of Peroxidase Isozyme during the Ripening of Tomato Fruits

Climacteric rise, ethylene production, peroxidase activity and its isozyme and their interrelationships during the ripening of tomato fruits have been studied. It was found' that there was parallelism between ethylene production and climacteric rise. The climacteric rise of tomato fruits was hastened by ethylene applied at the mature green stage. The ethylene production was inhibited by low oxygen and high carbon dioxide partial pressure. The peroxidase activity in the tomato fruits appeared to be different at three stages, higher in the half red fruits and lower in both green mature and fully red fruits. This activity was increased by ethylene and decreased by lower partial pres- sure of oxygen. The peroxidase isozymes sppeared also different at different stages of ripening. There were 4 bands in young fruits, 3 in green mature fruits, 5 in half red fruits and 3 in fully red fruits. After the application of ethylene to the tomato fruits, there appear one new band of peroxidase isozyme.     

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。     

吡虫啉对不同发育阶段蜘蛛酯酶同工酶活性的影响

本文采用聚丙烯酰胺凝胶电泳技术研究了吡虫啉对不同发育阶段星豹蛛酯酶同工酶活性的影响.结果表明,星豹蛛酯酶同工酶酶带可分为三个区域,其中EST-3区的酶带颜色最深,酶活性最强;EST-1区和EST-2区酶带颜色较浅,酶活性较弱.星豹蛛不同发育阶段酶带数由多到少的顺序为:雌成蛛＞卵袋＞雄成蛛＞若蛛,酶活性由强到弱的顺序为:...     

Developmental genetics of the esterase isozymes of Fundulus heteroclitus

The ontogeny of the esterase isozymes of the teleost, Fundulus heteroclitus, has been investigated. One group of esterase isozymes is present at all stages of development, whereas other esterase isozymes only very gradually appear at later stages of development, or abruptly appear at such dramatic developmental events as hatching. The ontogeny of these isozyme patterns is interpreted as the expression of differential regulation of separate esterase genes. The general pattern of teleost esterase gene activation is similar to that reported for birds and mammals. Allelic variation was detected at two of the esterase loci. On the basis of electrophoretic mobility, substrate specificity, inhibitor specificity, genetic variation, and ontogeny of esterases, there appear to be at least 15 different esterase isozymes, which constitute 6–8 groups, each of which is probably encoded in one or more genetic loci.This study was supported by NSF Grant GB 544OX to Professor C. L. Markert and an NSF Graduate Fellowship to G. S. Whitt.