33个黄花菜品种及3种野生黄花菜的过氧化物酶同工酶分析

１９８４－１９８７年先后全国１６个省区收集了３３个黄花菜（ＨｅｍｅｒｏｃａｌｌｉｓｃｉｔｒｉｎａＢａｒａｎｉ）品种和３个野生黄花菜品种。８７－９４年连续７年观察了各品种的主要形态特征,农艺性状。利用聚丙烯酰胺凝胶电泳技术分析比较了各品种的过氧化物酶同工酶谱。结果表明,凡形态上差异较大者,过氧化物酶谱同工酶谱差异也大;形态上无明显差异者,过氧化物酶同工酶谱也基本相同,表现了很强的对应性。各地栽培的黄花菜中普遍存在着同物异名或同名异物现象。收集的３３个黄花菜品种整理归并为２５个不同的品种;１７号早黄花（野生）、１９号四月花（栽培）均为北萱草（Ｈｅｍｅｒｏｃａｌｌｉｓｅｓｃｕｌｅｎｔｓ）,１８号野黄花为折叶萱草（Ｈｅｍｅｒｏｃａｌｌｉｓｐｌｉｃａｔａ）。为研究黄花菜品种的亲缘起源关系及进行杂交育种工作提供了一些理论依据。     

杉木不同无性系过氧化物酶同工酶的研究

本文对杉木（Ｃｕｎｎｉｎｇｈａｍｉａｌａｎｃｅｌｏａｔａ（Ｌａｍｂ．）Ｈｏｏｋ．）不同无性系及其不同株龄的叶和不同发育时期的雄球花进行了过氧化物酶同工酶检测。结果表明：杉木４１４号无性系叶和雄球花的过氧化物酶同工酶酶谱与其它无性系都具明显差异;同－无性系叶与雄球花的过氧化物酶同工酶酶谱存在显著差异;不同发育时期的雄球花的过氧化物酶同工酶存在顺序表达。作者认为：过氧化物酶同工酶作为种内分类的鉴定性状较为合适,而科、属、种的分类依据则应更侧重于形态特征。根据杉木４１４号无性系的过氧化物酶同工酶的特异性及其短叶、叶端钝圆和多雄花等特点,可以把它定为杉科杉属杉木物种的一个变型：多雄花杉木。     

番茄属(Lycopersicon)四个种的过氧化物酶同工酶分析

本文报告了应用连续浓度梯度聚丙烯酰胺凝胶电泳对番茄属Lyco-persicon的四个种:秘鲁番茄L.peruviaunm Mill.,多毛番茄L.hirsutum Humb.et Bonp,醋栗番茄L.pimpinellifolium(Jusl)Mill和普通番茄L.esculentum Mill.的86份材料,15个不同生育时期,不同器官以及同一器官的不同部位的过氧化物酶同工酶的分析结果。结果表明:L.Peruvianum的各个生育期和不同器官的过氧化物酶同工酶谱带叠加共有28条带,L.hirsutum有29条带,L.pimpinellifolium有28条带,L.esculentum有27条带。种间过氧化物酶同工酶谱型差异明显,种内不同生育期叠加总酶谱基本一致。在根、茎和叶中,这四个种的过氧化物酶同工酶酶谱和活性具有相似的生育期变化规律和器官分布规律.在果实发育过程中,种间过氧化物酶同工酶酶谱、活性及变化律规都不相同。本文还就同工酶谱型相似值的意义,野生资源及同工酶分析技术在番茄育种中的应用等问题进行了讨论。     

张杂谷苹果酸脱氢酶和过氧化物酶同工酶研究

以张杂谷‘3号’、‘5号’、‘6号’及其亲本为材料,通过聚丙烯酰胺垂直板凝胶电泳技术研究不同材料不同生育期苹果酸脱氢酶和过氧化物酶同工酶酶谱特性,同时对总酶活进行了测定,探讨谷子杂种优势的形成机理。结果表明:(1)张杂谷及其亲本叶片苹果酸脱氢酶共有5条酶带,有共同酶带和偏母本型酶带;过氧化物酶同工酶共有14条,属于共同酶带;从负极到正极苹果酸脱氢酶同工酶Rf分别为0.611、0.626、0.642、0.684和0.716,而过氧化物酶同工酶Rf分别为0.08、0.21、0.24、0.30、0.36、0.44、0.49、0.58、0.66、0.69、0.72、0.75、0.85和0.89。(2)杂种苹果酸脱氢酶同工酶和母本酶谱同型,过氧化物酶同工酶酶谱和父母本一致;供试品种苹果酸脱氢酶酶谱相对简单,酶带集中,而过氧化物酶同工酶酶带数量、活性和宽度差异性较大,酶谱比较复杂。(3)所有品种苹果酸脱氢酶总活性在抽穗期最高,过氧化物酶总活性在灌浆初期最高。杂种苹果酸脱氢酶活性在苗期和抽穗期表现出不同程度的超亲优势,而过氧化物酶在抽穗和灌浆初期均表现出不同程度的超亲优势,这可能与杂种优势有关。     

几种薏苡的过氧化物酶和多酚氧化酶同工酶分析

用聚丙烯酰胺凝胶电泳（PAGE）的方法,对薏苡,野生薏苡和水生薏苡的几个不同种质分别进行了过氧化物酶（POD）和多酚氧化酶（PPO）的同工酶酶谱分析,结果表明：POD和PPO含量丰富,活性强,酶谱复杂,酶带清晰,稳定,种间差异明显。结果可以作为薏苡研究种质亲缘关系的基础。     

欧李叶片过氧化物酶同工酶与叶果矿质元素含量变化的关系

以同一生境条件下113份燕山山脉野生欧李为试材,采用聚丙烯酰胺凝胶电泳方法,研究欧李叶片过氧化物酶(POD)同工酶与叶果中6种矿质元素的含量变化及其相关关系,为在酶水平上揭示欧李矿质营养代谢遗传差异提供理论参考。结果表明,欧李叶片过氧化物酶同工酶划分为7种酶谱类型,其中Rf 0.126和Rf 0.716酶带为共有的特征带,不同酶谱类型中单株个体间的相同酶带酶量大小以及叶果中的矿质元素含量存在明显差异,变异程度较大;相关分析结果显示,不同酶谱类型中相关酶带酶量大小与叶果中矿质元素含量变化存在显著或极显著的相关关系,表明欧李叶片过氧化物酶同工酶酶量大小变化在一定程度上反映矿质营养代谢的遗传差异和多样性。     

小麦与玉米及鸭茅状摩擦禾远缘杂交的胚发育过程中同工酶和蛋白质的变化

通过比较小麦与玉米及鸭茅状摩擦禾属间杂交获得的胚与小麦正常自交的胚之间在不同发育时期过氧化物酶和酯酶的同工酶谱,发现过氧化物酶同工酶表现出时空顺序的特异性变化。在同一发育时期,远缘杂交的具胚子房和无胚子房之间存在过氧化物酶同工酶谱的差异,这可能涉及到与胚发育相关的同工酶的出现。远缘杂交的具胚子房和正常自交的小麦子房之间也有一定的酶谱差异。同时,同一材料还表现出不同发育时期的过氧化物酶酶谱差别。在远缘杂交后的胚发育期间,酯酶同工酶的时空表达不如过氧化物酶显著。此外,对远缘杂交后的胚中的水溶性蛋白质进行了SD S-PAGE分析,初步的分析结果表明,可能存在与胚发育相关的蛋白质。     

甘薯属植物过氧化物酶同工酶分析

采用垂直平板聚丙烯酰胺凝胶电泳技术,对23份甘薯属不同倍性材料进行过氧化物酶同工酶酶谱分析。初步结果表明,过氧化物酶同工酶酶带数目与材料倍性无明显相关性;二倍体或四倍体野生种的种间酶谱差异显著;六倍体野生种不同株系间以及六倍体栽培种甘薯的不同品种间酶谱差异较小;但栽培种甘薯与六倍体野生种I.trifida(6x)、四倍体野生种I.littoralis(4x)以及二倍体野生种I.trifida(2x)的酶谱有4条明显共同标记带,表明其间有一定亲缘关系。     

离子注入对萝卜过氧化物酶、淀粉酶和蛋白酶同工酶的影响

研究了氮离子和氩离子注入萝卜种子对萝卜幼苗蛋白含量、过氧化物酶活性及过氧化物酶、淀粉酶和蛋白酶同工酶的影响。结果表明 :离子注入后 ,减低萝卜过氧化物酶活性和蛋白含量。萝卜不同生长时期同工酶变化不一样。在子叶时期 ,过氧化物酶同工酶谱带无明显变化 ,淀粉酶同工酶有酶带的消失 ;而在真叶时期 ,过氧化物酶在负极区减少一条酶带 ,Rf为 0 .2 2 ,正极区增加一条酶带 ,Rf为 0 .6 ,且随剂量增加 ,酶带着色增强 ;淀粉酶同工酶在注入剂量为 5× 10 5N+ / cm2 )时 ,有同工酶带增加 ,Rf为 0 .6 1。低剂量时蛋白酶活性增强 ,谱带增多 ,大剂量则减弱。因此 ,N+和 Ar+注入后 ,可影响萝卜过氧化物酶和淀粉酶的表达及蛋白质的合成或降解。     

萝卜贮藏期过氧化物酶活性及同工酶谱研究

为了研究萝卜贮藏期过氧化物酶的活性变化和作用 ,利用愈创木酚法和聚丙烯酰胺凝胶电泳方法检测在贮藏期萝卜过氧化物酶的活性及同工酶谱 ,实验结果表明 :不同品种的萝卜在不同贮藏时期其过氧化物酶活性及同工酶谱存在差异。贮藏时间越长过氧化物酶活性越高 ,同工酶谱酶带增多。萝卜色素含量越高 ,POD活性也越高 ,因此 ,在萝卜贮藏期有可能过氧化物酶在清除细胞内H2 O2 方面起主要作用     

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.     

High resolution of peroxidase-indoleacetic Acid oxidase isoenzymes from horseradish by isoelectric focusing

Several improved techniques for isoelectric focusing of isoenzymes in polyacrylamide gel slabs were developed. Using these techniques, three commercial sources of horseradish peroxidase were each examined with three commercial sources of carrier ampholytes to determine their respective isoenzyme profiles.     

Regulation of synthesis of nitrite reductase in pea leaves: in-vivo and in-vitro studies

The cellular location of three peroxidase isoenzymes (PRX) in mature leaf tissue of Petunia and their affinity for Concanavalin A-Sepharose were investigated. The isoenzymes PRXa, PRXb and PRXc were identified by their positions in starch-gel zymograms. The fast-moving anodic and cathodic peroxidase bands, the isoenzymes PRXa and PRXc respectively, were the most active peroxidases in extracellular extracts. The molecular forms of PRXa showed a tissue-specific distribution between midrib and remaining leaf tissue. An intermediate-moving anodic peroxidase band, the isoenzyme PRXb, was the most active peroxidase released after extraction of isolated mesophyll protoplasts. Small amounts of the peroxidase isoenzymes were present in cell-wall-bound fractions. Incubation of a crude protein fraction with Concanavalin A-Sepharose showed that the isoenzyme PRXb bound more firmly to Concanavalin A-Sepharose than the isoenzymes PRXa and PRXc, of which only one molecular form bound partly. The results are discussed with respect to a possible function of one of the peroxidase isoenzymes, and a possible role of oligosaccharide chains in determining the cellular location of plant peroxidases is suggested.Abbreviations Con A Concanavalin A - PRX peroxidase (isoenzyme)     

Proton nuclear magnetic resonance spectroscopy of horseradish peroxidase isoenzymes: correlation of distinctive spectra with isoenzyme specific activities

High-resolution proton NMR spectra are reported for the paramagnetic ferric native and cyano complexes of the five major horseradish root peroxidase (HRP) isoenzymes (A1, A2, A3, B, and C). Axial imidazole resonances are observed in the native and cyano-complex spectra of all the isoenzymes, thus indicating the presence of a common axial histidine ligand. Proton NMR spectra outside the usual diamagnetic region are identical for sets of A1 and A2 isoenzymes and for the B and C isoenzyme set. Variation in heme residue chemical shift positions may be controlled in part by porphyrin vinyl side chain-protein interactions. Diverse upfield spectra among the isoenzymes reflect amino acid substitutions and/or conformational differences near the prosthetic group, as signals in this region must result from amino acid residues in proximity to the heme center. Acid-base dependence studies reveal an "alkaline" transition that converts the native high-spin iron (III) porphyrin to the low-spin state. The transition occurs at pH 9.3, 9.4, 9.8, and 10.9 for respective HRP A1, A2, A3, and C isoenzymes, respectively. Significantly, this ordering also reflects specific activities for the isoenzymes in the order A1 = A2 greater than A3 greater than B = C. Identical proton NMR spectra for A1/A2 and B/C isoenzyme sets parallel equivalent specific activities for members of a particular set. Proton NMR spectra thus appear to be highly sensitive to protein modifications that affect catalytic activity.     

苦豆子碱和烟碱的联合杀蚜作用

【目的】以豆蚜Aphis craccivora Koch为试虫,对苦豆子碱和烟碱的联合杀蚜活性进行了测定,以期为开发新型植物源杀蚜剂奠定基础。【方法】单体化合物对豆蚜的毒力测定采用微量点滴法;制剂对豆蚜的毒力测定采用波特喷雾法;田间药效试验参照国家标准进行,其中施药方法采用常量喷雾法;制剂加工及化合物之间的联合杀蚜作用测定均采用常规方法。【结果】苦豆子碱和烟碱混用具有明显的杀蚜增效作用,在最佳质量配比（苦豆子碱∶烟碱=13∶1）下,共毒系数（common toxicity coefficient, CTC）值达185.82。结合对溶剂、乳化剂等助剂的筛选,研制出15.7% 苦豆子碱·烟碱环保型乳油,配方为：苦豆子碱（14.6%）、烟碱（1.1%）、环保溶剂（74.3%）、乳化剂（10%）。质量检测结果表明该制剂各项指标均达到商品农药的要求。大田药效试验表明,该制剂在质量浓度为0.354 g/L时,常量喷雾,药后7 d对月季长管蚜Macrosiphum rosirvorum的防效仍为85.91%。【结论】苦豆子碱和烟碱混用具有显著的杀蚜相互增效作用,具有进一步开发潜力。     

Purification and characterization of Cu-Zn superoxide dismutases from mungbean (Vigna radiata) seedlings

Mungbean contains three isoenzymes of superoxide dismutase designated isoenzyme I, II and III. The two cytosolic superoxide dismutases (I and II) were purified to homogeneity by ammonium sulphate fractionation, ion exchange chromatography on diethylaminoethyl cellulose, gel filtration and preparative polyacrylamide.gel electrophoresis. The molecular weights of isoenzyme I and isoenzyme II were determined to be 33,000 and 31,600 respectively. The subunit molecular weight was approximately 16,000 indicating that the isoenzymes contained two identical subunits. The ultra-violet absorption spectra revealed a maximum at 258–264 nm for the two isoenzymes. Superoxide dismutase I and II were inhibited to different extents by metal chelators. Isoenzyme I was more sensitive to inhibition by cyanide and azide, while isoenzyme II was more susceptible to inhibition by diethyldithiocarbamate ando-phenanthroline. Both the isoenzymes exhibited similar denaturation profiles with heat, guanidinium chloride and urea. The denaturation with urea and guanidinium chloride was reversible. The two copper-zinc enzymes were more stable towards thermal inactivation compared to manganese and iron superoxide dismutases from other sources. The results indicate that the two isoenzymes differ from each other only with respect to charge and sensitivity towards metal chelators.     

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.     

Separation of rat lactate dehydrogenase isoenzyme C4 from other isoenzymes by affinity and ion-exchange chromatography.

Lactate dehydrogenase C, an isoenzyme composed of C polypeptide subunits and found only in mature testes and spermatozoa, differs kinetically, chemically and immunologically from the five common isoenzymes of lactate dehydrogenase, each of which is a tetramer of A and/or B subunits. In the rat lactate dehydrogenase C exists in two molecular forms, isoenzymes C4 and A1C3. In addition to these two forms of lactate dehydrogenase C, rat testicular homogenate contains all the five isoenzymes of A and B type. Purification of isoenzyme C4 requires its separation from the other six isoenzymes, of which isoenzymes A1C3 and A3B1 are the most difficult ones to separate. In the present study isoenzyme A3B1, along with other enzymes, was separated from isoenzyme C4 by AMP-Sepharose chromatography by using a gradient of increasing concentration of NAD+-pyruvate adduct. In the next step, isoenzyme A1C3 was separated from isoenzyme C4 by DEAD-cellulose chromatography, resulting in a pure lactate dehydrogenase isoenzyme C4 preparation.     

Isoenzymes of horse liver alcohol dehydrogenase active on ethanol and steroids. cDNA cloning, expression, and comparison of active sites.

Horse liver alcohol dehydrogenase occurs as isoenzymes: E is active on ethanol but not steroids; S is active on ethanol and steroids. The cDNAs for these isoenzymes were cloned; both were 1.8-kilobase long and contained complete coding sequences. Both enzymes were expressed in Escherichia coli, and the purified proteins had properties similar to those of the natural enzymes. The amino acid sequence deduced from the open reading frame of the E-type cDNA agreed with the amino acid sequence of the E isoenzyme determined by protein sequencing and x-ray crystallography. When compared with the E-type cDNA, the coding region of the S-type cDNA contains 24 substitutions and 3 deletions, giving rise to an amino acid sequence for the S. isoenzyme that differs from that of the E isoenzyme at 10 positions: nine conservative substitutions and one deletion, of Asp-115. These changes can be accommodated in the three-dimensional structure of the E isoenzyme, and models of the E and S isoenzymes complexed with a 3 beta-hydroxy-5 beta-steroid were built. The modeling shows that Leu-116 apparently sterically hinders binding of steroids in the E isoenzyme, and deletion in the S isoenzyme of Asp-115 moves Leu-116 and relieves the hindrance. The human gamma and rat liver enzymes are also active on steroids, but they have a different constellation of amino acid residues in the substrate pocket. Thus, there are multiple bases for the activity on steroids.     

The development of glutathione S-transferase and glutathione peroxidase activities in human lung

The development of glutathione S-transferase and glutathione peroxidase activities has been studied in human lung cytosols. Whilst no clear change in glutathione peroxidase activity was identified, expression of the acidic glutathione S-transferase isoenzyme decreased markedly after 15 weeks of gestation so that at birth the level of activity of this isoenzyme was only about 20% of that in samples obtained during the first trimester. Basic glutathione S-transferase isoenzymes were weakly expressed during development and usually comprised less than 10% of cytosolic activity. Ion-exchange studies identified several basic isoenzymes that may correspond to the alpha, beta, gamma, delta and epsilon set previously identified in liver. Weak expression of apparently near-neutral isoenzymes was also detected; they were detected in only a few cytosols.