等位酶分析的遗传学基础（续）

在等位酶分析中,对酶谱的正确解释是获取遗传学资料的基础,酶谱作为酶基因的表现型,是由该种酶蛋白质的四级结构情况（亚基的数目）、在亚细胞分室中的分布（位点的数目）以及所分析样品的倍性和基因型的情况所决定的。术语“酶型”被建议用来记录和描述各种情况下酶谱的不同。在进行分子系统学研究中,如果直接把酶谱上带的数目的多少作为遗传多样性大小的指标,或把带的多少及迁移率的大小作为数量性状进行聚类和分枝分析将会得出非常错误的结论。     

半夏种内不同居群酯酶和超氧化物歧化酶同工酶酶谱特征分析

对栽培在同一生境下的16个半夏〔Pinellia ternata（Thunb.）Breit.〕居群同一生长期叶片中酯酶（EST）和超氧化物歧化酶（SOD）同工酶酶谱进行了比较分析,结果表明：EST和SOD同工酶酶谱在各居群间,甚至在同一居群内除少数共有的特征谱带外存在着较为明显的频率差异。EST同工酶在半夏各居群中最多可显示12条谱带,其中在慢区的第5（弱带）和快区的第10（强带）2条谱带为各居群所共有的特征谱带;SOD同工酶在半夏各居群中最多显示9条谱带,其中在慢区的第2（弱带）、快区的第5（强带）和第9（强带）3条谱带为各居群所共有的特 征谱带。     

木耳栽培菌株酯酶同工酶的酶谱多样性研究

采用聚丙烯酰胺凝胶电泳技术对木耳（Ａｕｒｉｃｕｌａｒｉａ ａｕｒｉｃｕｌａ （Ｈｏｏｋ）Ｕｎｄｅｒｗ）１０个栽培菌株酯酶同工酶的酶谱多样性进行了研究。１０个供试菌株的幼龄菌丝（７ｄ）中仅检测到２５条酶带,各个菌株分别具有２～３条酶带,１０个菌株仅有２种酶谱类型;老龄菌丝（７２ｄ）中共检测到４４条酶带,各个菌株分别具有３～６条酶带,１０菌株共有８种酶谱类型。研究表明,木耳双核体菌丝中某些酯酶同工酶棋     

木耳栽培菌株酯酶同工酶的酶谱多样性研究

采用聚丙烯酰胺凝胶电泳技术对木耳（Ａｕｒｉｃｕｌａｒｉａａｕｒｉｃｕｌａ（Ｈｏｏｋ）Ｕｎｄｅｒｗ）１０个栽培菌株酯酶同工酶的酶谱多样性进行了研究。１０个供试菌株的幼龄菌丝（７ｄ）中仅检测到２５条酶带,各个菌株分别具有２～３条酶带,１０个菌株仅有２种酶谱类型;老龄菌丝（７２ｄ）中共检测到４４条酶带,各个菌株分别具有３～６条酶带,１０菌株共有８种酶谱类型。研究表明,木耳双核体菌丝中某些酯酶同工酶基因位点在一定的发育时期才开始表达,老龄菌丝酯酶同工酶酶谱在木耳菌株鉴别和遗传育种研究中具有更大的应用价值。     

蝗总科部分种类等位基因酶的比较研究

用水平淀粉凝胶电泳技术对采自山西太原黄陵、山西临猗伍姓湖及山西雁门关两科4种蝗虫的4个等位基因酶位点的基因频率进行了比较研究,并用BIOSYS-Ⅱ软件进行结果分析。结果表明：所研究种群的苹果酸脱氢酶（MDH）都存在两个位点,中华稻蝗的MDH-1还存在两条亚带。在所研究种群中的MDH-1图谱中,一个中等迁移率的谱带存在于所研究的4个种群中。东亚飞蝗在乳酸脱氢酶（LDH）和苹果酸酶（ME）中存在两个固定的等位基因,同时黄胫小车蝗在MDH-1中存在一个固定的等位基因,在MDH-2中存在一个独特的等位基因。在所有4个种群中,中华稻蝗遗传多样性水平最高（每个位点的等位基因数为3个,He=0.220）,黄胫小车蝗遗传多样性水平最低（每个位点的等位基因数为1.5个,He=0.013）。除中华稻蝗的MDH-1和LDH处于哈代-温伯格平衡,其余种群的4个位点的等位基因频率均不同程度的偏离哈代-温伯格平衡。等位酶数据表明这4个种群在系统发育关系方面是相近的,但在遗传多样性水平上却不同。     

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。     

辣椒细胞质雄性不育系的3种同工酶分析

以辣椒细胞质雄性不育系21A及其同核异质保持系21B为试验材料,比较分析两系雄配子发育过程中酯酶（EST）、谷氨酸脱氢酶（GDH）和苹果酸脱氢酶（MDH）同工酶的表达特征。结果表明：幼叶和花蕾的EST同工酶酶谱在谱带数目和酶带强弱上存在时空表达差异,并且随着雄配子发育的进行,保持系21B从中花蕾至特大花蕾比不育系21A多1条清晰的谱带（EST3e）,其差异表达发生在细胞学上观察到的败育时期之前;在GDH同工酶中,保持系21B从大花蕾至特大花蕾比不育系多6条谱带（GDH,和GDH1／2）,酶谱差异表达时期与细胞学上观察到的败育时期一致;而在MDH同工酶中,不育系21A和保持系21B的幼叶和各级花蕾的酶谱在谱带数目和谱带强弱上均没有明显差异。     

芒果属植物过氧化物酶同工酶的研究

采用聚丙烯酰胺凝胶电泳法,测定了芒果属１７个种群近缘植物过氧化物酶同工酶,并根据酶谱分布及酶谱特征,结合不同的表现形式,分析了属内种间的亲缘关系。结果表明,异（同）地种间、不同器官酶谱数目、酶含量、迁移率（Ｒｆ）、酶活性及分布特征均有不同程度的差异,可以作为芒果属种间亲缘关系的生化遗传指标。     

IPsaur-ipt基因转化番茄的过氧化物酶同工酶分析

将整合在Ｔｉ质粒上的Ｐｓａｕｒ基因（生长素调节基因和细胞激动素合成酶基因的融合基因）通过花粉管通道法导入番茄品种中蔬四号、强力米寿、粉红甜肉、ＵＳ—Ⅰ、ＵＳ—Ⅱ中。对Ｄ１代转基因番茄用聚丙稀酰凝胶电泳方法进行过氧化物酶同工分析。结果表明,转基因番茄与对照植株的过氧化物酶同工酶谱带有明显差异,谱带多少和强弱都有所不同,而且相应地出现了植株形态学上的差异。间接验证了外源ＤＮＡ的导入。说明同工酶分析方法和田间形态学检测可作为转基因植株早期筛选的有效方法     

高背银鲫、大口胭脂鱼及胭脂鲫(F1)不同组织的同工酶的表型分析

本文对高背银鲫、大口胭脂鱼及胭脂鲫（F1）不同组织的脂酶（EST）、乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）、超氧化物歧化酶（SOD）四种酶的同工酶的表型进行了分析,结果表明：在肝脏中胭脂鲫EST多一条谱带,而在性腺中,LDH、MDH、SOD同工酶谱带,胭脂鲫与父母本均存在一定的差异。     

Distribution of aspartate aminotransferase activity in yeasts, and purification and characterization of mitochondrial and cytosolic isoenzymes from Rhodotorula minuta [corrected

The distribution of aspartate aminotransferase activity in yeasts was determined. The number of species of the enzyme in each yeast was determined by zymogram analysis. All the yeasts, except for the genus Saccharomyces, showed two or three activity bands on a zymogram. From among the strains, Rhodotorula minuta [corrected] and Torulopsis candida were selected for examination of the existence of yeast mitochondrial isoenzymes, because these strains showed two clear activity bands on the zymogram and contained a high amount of the enzyme. Only one aspartate aminotransferase was purified from T. candida: the component in the minor band on the zymogram was not an isoenzyme of aspartate aminotransferase. On the other hand, two aspartate aminotransferases were purified to homogeneity from R. minuta [corrected]. The components in the main and minor activity bands on the zymogram were identified as the mitochondrial and cytosolic isoenzymes, respectively, in a cell-fractionation experiment. The enzymatic properties of these isoenzymes were determined. The yeast mitochondrial isoenzyme resembled the animal mitochondrial isoenzymes in molecular weight (subunits and native form), absorption spectrum, and substrate specificity. The amino acid composition was closely similar to that of pig mitochondrial isoenzyme. Rabbit antibody against the yeast mitochondrial isoenzyme, however, did not form a precipitin band with the pig mitochondrial isoenzyme.     

角类肥蛛体躯不同部位酯酶同工酶的比较分析

运用聚丙烯酰胺凝胶电泳的方法对角类肥蛛(Lariniodes cornuta)头胸部和腹部的酯酶同工酶酶谱进行了比较分析。结果表明,角类肥蛛的酯酶是单体酶,头胸部和腹部的酯酶酶谱差异显著。腹部的酯酶呈现4个位点:Est-1、Est-2、Est-3、Est-4。Est-1和Est-4位点为纯合基因型,Est-2和Est-3位点为杂合基因型。头胸部的酯酶仅表现出2个位点:Est-2和Est-3,且这2个位点是纯合基因型。不同个体之间头胸部的酯酶没有明显差异,Est-2b和Est-3a可以作为鉴别角类肥蛛的特征酶带;腹部的酯酶则存在明显的个体差异,在Est-2和Est-3位点的基因杂合度为h2=h3=0·4779。由此可见,酯酶同工酶可以作为角类肥蛛遗传变异的分子标记,是研究个体间遗传差异、居群的遗传结构以及种间进化关系的基础。     

Study on the Esterase Isozyme from the Related Citrus Plants

In this experiment, the polyacrylamide gel electrophoresis is adopted to test the esterase isozyme of seeds and seedling of the fourteen species of related Citrus plants. The relationship with each other is analysed according to the resutls of zymogram and scanning chart. The data show that there are differences of various degrees in the number of the isozyme bands, the enzyme activity, the migration of enzymes and their chromatogram scanning. The isozyme of the same species has different forms of expression in the different period of development. The esterase isozyme zymogram of the Citrus seed is generally stable. It can be used as the biochemical and genetical index to detemine the genetic ralationship of Citrus.     

柑桔近缘植物酯酶同工酶的研究

本文用聚丙烯酰胺凝胶电泳测定了柑桔近缘植物14个种群的种子及幼苗的酯酶同工酶,根据酶谱及扫描图的异同,分析了彼此的亲缘关系,试验结果表明,柑桔近缘植物种属间的酯酶同工酶的酶带数目,酶活性,迁移率及酶谱扫描均有不同程度的差异,同一品种不同发育时期的同工酶也具有不同表现形式,特别是柑桔种子的酯酶同工酶谱一般较稳定,可以作为柑桔亲缘关系的生化遗传指标。     

Differentiation of Physiologic Races of the Rice Blast Fungus, Pyricularia oryzae Cav. by PAGE-electrophoresis

Eighty-three isolates of the rice blast fungus (Pyricularia oryzae) were tested with respect to genetic diversity and the possibility of race differentiation by electrophoresis. The fungus was genetically very heterogeneous. The isolates were differentiated into 6 races by pathogenicity on race differential varieties. There was little correlation between pathogenicity and zymogram types of one particular enzyme such as esterase, phosphatase or catalase. The isolates were divided into 14 groups by the combination of the zymogram types of the three enzymes. The isolates in the same group showed similar pathogenicity. A new method is proposed which differentiates the blast fungus races by the combination of zymogram type of enzymes. The details, are discussed.     

Isozyme Studies in Predicting the Potential Yield of Heterosis in Maize

Electrophoretic isozyme zymogram patterns of peroxidase (POD), cytochrome oxidase (COD), esterase (Est), α-amylase (α-Amy), catalase (Cat), Glutamate dehydrogenase (GDH), Malate dehydrogenase (MDH), superoxide dismutase (SOD), acid phosphatase (Acp), etc. obtained from 108 maize inbred lines and their 199 hybrids were analyzed. The soluble protein patterns from these materials were tested as well. The authors had probed into the relationship between the index of zymogram difference and potential yield of heterosis in maize. The results indicated that hybrids from parents which showed high zymogram difference index could produce high heterosis and those from parents which had low zymogram difference index could also produce high heterosis as well. But hybrids from the sister lines which had lower zymogram difference index could only produce lower heterosis. There was no significant statistical difference between isozyme zymogram difference index and potential yield of heteroSis on a genetic background with complex combinaton.     

三种杜拉属（Drawida）蚯蚓的酯酶同工酶的初步研究

应用聚丙烯酰胺凝胶电泳的方法，对寡毛纲杜拉属三种蚯蚓进行酯酶同工酶酶谱分析。其结果表明，同属蚯蚓具有几条相似的酶谱区带，而不同种蚯蚓又具有各自独特的酶谱带，种间酶谱区带数目、泳动率及染色强度都有明显区别。雅和杜拉蚓酶谱图和天锡杜拉蚓相似，而与管状杜拉蚓相差较远，表明同属蚯蚓在进化水平上的亲缘关系。     

Specific Isozyme Marker of a Virus Resistant Barley Derived from Interspecies Cross Between Hordeum vulgare and H. bulbosum

A diploid barley cultivar "Supi 1" was crossed with a tetraploid Hordeum bulbosum “GBC141” to transfer the disease resistant traits. Eleven viable triploid F1 plants were produced by means of embryo rescue technique. The resulting triploid hybrids were backcrossed to diploid barley, and seven BC1 plants were obtained. One of the BC1 plants exhibited barley yellow mosaic virus (BaYMV) resistance when grown in the diseased nursery. Isozyme analysis of H. vulgate, H. bulbosum and their backcross hybrids were made via slab polyacrylamide gel electrophoresis technique. The primary results showed that zymogram variation could be obviously found between diploid barley "Supi 1' and tetraploid H. bulbosurn "GBC141”. A peroxidase isozyme (Rf=0.47) from H. bulbosum was detected in the peroxidase isozyme zymogram of young roots of backcross hybrid BC1-2. This peroxidase isozyme was related to the BaYMV resistance but the linkage relation will be determined by the genetic analysis of the F2 population in the future. The BaYMV resistant line of the backcross with isozyme marker is the important resource of barley disease-resistant breeding.     

Maize starch-branching enzyme isoforms and amylopectin structure. In the absence of starch-branching enzyme IIb, the further absence of starch-branching enzyme Ia leads to increased branching

Previous studies indicated that the deficiency of starch-branching enzyme (SBE) Ia in the single mutant sbe1a::Mu (sbe1a) has no impact on endosperm starch structure, whereas the deficiency of SBEIIb in the ae mutant is well known to reduce the branching of starch. We hypothesized that in maize (Zea mays) endosperm, the function of SBEIIb is predominant to that of SBEIa, and SBEIa would have an observable effect only on amylopectin structure in the absence of SBEIIb. To test this hypothesis, the mutant sbe1a was introgressed into lines containing either wx (lacking the granule-bound starch synthase GBSSI) or ae wx (lacking both SBEIIb and GBSSI) in the W64A background. Both western blotting and zymogram analysis confirmed the SBEIa deficiency in sbe1a wx and sbe1a ae wx, and the SBEIIb deficiency in ae wx and sbe1a ae wx. Using zymogram analysis, no pleiotropic effects of sbe1a genes on SBEIIa, starch synthase, or starch-debranching enzyme isoforms were observed. High-performance size exclusion chromatography analysis shows that the chain-length profiles of amylopectin as well as beta-limit dextrin were indistinguishable between wx and sbe1a wx, whereas significant differences for both were observed between ae wx and sbe1a ae wx, suggesting an effect of SBEIa on amylopectin biosynthesis that is observable only in the absence of SBEIIb. The amylopectin branch density and the average number of branches per cluster were both higher in endosperm starch from sbe1a ae wx than from ae wx. These results indicate possible functional interactions between SBE isoforms that may involve enzymatic inhibition. Both the cluster repeat distance and the distance between branch points on the short intracluster chains were similar for all genotypes however, suggesting a similar pattern of individual SBE isoforms in cluster initiation and the determination of branch point location.