【作者】 程汉良；

【导师】 夏德全； 吴婷婷；

【作者基本信息】 南京农业大学 ， 水生生物学， 2006， 博士

【摘要】 1．4种帘蛤科贝类同工酶研究 采用不连续垂直板聚丙烯酰胺凝胶电泳技术，对文蛤(Meretrix meretrix L．，1758)、青蛤(Cyclina sinensis G．，1791)、硬壳蛤(Mercenaria mercenaria L．，1758)和菲律宾蛤仔(Ruditapes philippinarum A．，1850)的4种组织(肝、鳃、后闭壳肌和外套膜)的酯酶(EST)、苹果酸脱氢酶(MDH)和超氧化物歧化酶(SOD)同工酶进行了研究。结果表明，不同帘蛤的同工酶谱有明显的差异；同种帘蛤不同组织间的EST和MDH同工酶有一定的组织特异性；EST和MDH同工酶位点丰富，可用于不同物种的鉴定；肝、鳃EST和闭壳肌MDH酶活性较高，是进行同工酶分析的理想材料。 对硬壳蛤5种组织(足、外套膜、闭壳肌、鳃和肝)的酯酶(EST)、苹果酸脱氢酶(MDH)和乳酸脱氢酶(LDH)同工酶进行了比较研究，并对酶谱表型及位点表达进行了分析。结果表明，硬壳蛤上述组织的EST和MDH同工酶存在不同程度的组织特异性，而LDH没有组织特异性。在EST酶谱中，肝组织染色最深，说明其EST活性最高，这与肝脏是重要的消化腺和解毒器官相适应。在MDH酶谱中发现了s-MDH和m-MDH所形成的谱带，闭壳肌的MDH谱带染色很深，这与肌肉需要由三羧酸循环提供大量ATP相适应。 2．4种帘蛤群体遗传多样性和种间关系及文蛤4个地理种群遗传结构的fAFLP分析 利用荧光标记扩增片段长度多态性(fAFLP)技术对4种帘蛤(文蛤、青蛤、菲律宾蛤仔和硬壳蛤)的群体遗传多样性和种间关系进行了研究。选择EcoR Ⅰ／Mse Ⅰ进行酶切，使用6个E+3／M+3引物组合进行扩增，共获得1096个位点，多态位点比率95.1％，片段长度50-456bp。其中，文蛤、青蛤、菲律宾蛤仔和硬壳蛤分别得到681、715、702和694个位点，各物种相应的多态位点比率为76.8％、81.7％、83.0％和75.1％，得到17个物种特异性位点，可作为4物种特征标记。分析了群体遗传相似系数和遗传多样性指数以及种间遗传相似系数。结果表明，硬壳蛤群体遗传相似系数最高(0.6709)，遗传多样性指数最低(0.2360)；菲律宾蛤仔群体遗传相似系数最低更多还原

【Abstract】 1. Study on the isozymes of four clam species in Veneridae (Mollusca: Bivalvia)The isozymes (EST, MDH and SOD) from four different tissues (liver, gill, adductor muscle, mantle) of four clams species (Meretrix meretrix; Cyclina sinensis; Ruditapes Philippinarum and Mercenaria mercenaria) in Veneridae were analyzed by vertical polyacrylamide gel electrophoresis (PAGE). The result showed that there was obvious diversity in the isozyme phenotypes among four clams in Veneridae in all tissues inspected, and among four tissues in same species. EST and MDH showed more electrophoresis patterns, so they could be applied as markers to identify the different species in Veneridae. Liver and gill are better material for EST isozyme analysis and adductor muscle for MDH, because of the activity of enzyme is higher than other tissues.The electrophoretograms of isozymes (EST, MDH and LDH) from five different tissues (foot, mantle, adductor muscle, gill, liver) of M. mercenaria were analyzed. The results showed that there are some differences in the isozyme phenotypes of EST and MDH among five tissues, but no difference was found in LDH. The electrophoresis patterns of EST and MDH had a tissue-specific distribution. The physiological significance in relation to the tissue-specific distribution of the isozymes was discussed. And we found the gene of s-MDH and m-MDH in adductor muscle of M mercenaria.2. Fluorescent amplified fragment length polymorphism analysis of genetic diversity and species relationships of four clam species and population genetic structure of Meretrix meretrix in family Veneridae (Mollusca: Bivahia)Fluorescent amplified fragment length polymorphism (fAFLP) method was used to study the genetic diversity and species relationships of four species in family Veneridae. Digestion with enzymes EcoR I /Mse I and amplification with six E+3/M+3 primer更多还原