【作者】 王昭萍； 郭希明； 李赟； 于瑞海； 田传远； 王如才；

【Author】 WANG Zhaoping~1 GUO Ximing~2 LI Yun~1 YU Ruihai~1 TIAN Chuanyuan~1 WANG Rucai~1 1.Mariculture Research Laboratory,Ocean University of Qingdao,Qingdao 266003; 2.Haskin Shellfish Research Laboratory,Rutgers University,Pot Norris,NJ 08349,USA

【机构】 青岛海洋大学教育部海水养殖重点实验室； Haskin Shellfish Research Laborstory,Rutgers University；

【摘要】 采用水平淀粉凝胶电泳方法对细胞松弛素 B 抑制第二极体产生的三倍体（简称3nCB）和二倍体与四倍体杂交产生的三倍体（简称3nDT）太平洋牡蛎及其对照群体（简称2n）进行了同工酶研究。记录了 Aat₂、Ah₁、Ah₂、Idh 和 Lap 5个多态位点的基因型。二倍体与三倍体各多态位点的电泳酶带基本上按照预期模式表达。Aat₂、Idh 为二聚体酶,纯合子仅表现出一条酶带,杂合子则表现出3条酶带。2n 组中,3条酶带的相对染色强度为1：2：1,3nCB 和3nDT 三倍体中则为4：4： 1。Ah₁、Lap 为单体酶,纯合子表现出一条酶带,杂合子表现出2条酶带。2n 组中两条酶带的相对染色强度为1：1．三倍体中两条酶带的基因剂量为2：1。在 Ah₂位点上检测到3个不同的等位基因,纯合子也表现出1条酶带,杂合子在2n 对照组中表现为2条酶带,基因剂量为1：1；在三倍体中表现为2条或3条酶带,具2条酶带者相对染色强度为2：1,具3条酶带者相对染色强度为1： 1：1。根据杂合位点上各等位基因表达的剂量不同或酶带数目不同,可以区分出二倍体与三倍体, 特别是 Ah₂位点上3条酶带的出现可以准确地判断三倍体个体。通过比较流式细胞术和同工酶电泳对3nCB 群体倍性的分析结果表明,淀粉凝胶电泳方法是测定太平洋牡蛎多倍体的一种较有效的方法,三倍体检出率较高（96％～98％）。更多还原

【Abstract】 Five polyrnorphic loci were encoded in diploid（2n）and triploid Pacific oysters produced by inhabiting polar body Ⅱ with CB（3nCB）and mating diploid and tetraploid（3nDT）by starch electrophoresis:monomers Ah_1～2, Lap,and dimers Aat₂ and Idh₁.Electrophoretic examination indicated that diploids and triploids were separable ac- cording to the expected banding patterns:dimeric enzymes show one band at homozygous loci and 3 bands at het- erozygous loci.Heterozygous phenotypes exhibit staining intensities in ratio 1:2:1 in diploids,and 4:4:1 in triploids,respectively.Monomeric enzymes show one band at homozygous loci and two bands at heterozygous loci in ratio 1:1 in diploids and 2:1 in triploids,respectively.Three alleles were encoded at Ah₂.Heterozygotes at locus Ah₂ exhibited 2bands in ratio 1:1 in diploids,and 2 or 3 bands in ratio 2:1 or 1:1:1 in triploids.According to the relative staining intensities or the numbers of the component bands of heterozygous phenotype,triploids can be dis- tinguished from diploids,especially the occurrence of 3 bands of Ah₂ locus tell triploids from diploids precisely.Direct comparison of the results from flowcytometry and electrophoretic assays suggest that electrophoresis was an efficient （96%～98%）means of ploidy analysis in Pacific oyster.更多还原