【作者】 王昭萍； 郭希明； 李赟； 于瑞海； 田传远； 王如才；

【Author】 WANG Zhaoping, GUO Ximing, LI Yun, YU Ruihai, TIAN Chuanyuan, WANG Rucai1. Mariculture Research Laboratory, Ocean University of Qingdao, Qingdao 266003;2. Haskin Shellfish Research Laboratory, Rutgers University, Por 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 Laboratory,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Lap5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,纯合于仅表现出一条酶带,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 polymorphic 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 Ah1~2, Lap, and dimers Aat2 and Idh1. Electrophoretic examination indicated that diploids and triploids were separable according 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 Ah.2 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 distinguished 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% ?8% ) means of ploidy analysis in Pacific oyster.更多还原