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微卫星评价栉孔扇贝雌核发育二倍体纯合性 被引量:2

Assessment of homozygosity in gynogenetic diploid using microsatellite markers in scallop,Chlamys farreri
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摘要 采用筛选获得的5对多态性微卫星引物对栉孔扇贝(Chlamys farreri)减数分裂雌核发育家系A和家系B、有丝分裂雌核发育家系A和家系B各30个个体、双亲及对照组40个个体进行了遗传变异分析,并对减数分裂和有丝分裂雌核发育后代的纯合性进行了比较。电泳结果表明,5对微卫星引物在减数分裂雌核发育和有丝分裂雌核发育个体中均能稳定重复地扩增出相应的序列;各雌核发育家系中均有部分个体出现父本基因,表明精子遗传物质失活不彻底,雌核发育组中存在正常受精个体;有丝分裂雌核发育二倍体在所有检测位点全部纯合,减数分裂雌核发育二倍体在部分位点纯合,但未发现在所有位点全部纯合的个体,在CFMSP007、CFMSP075、CFMSM009、CFMSM014和CFMSM020位点,杂合子比例分别为0.3400、0.3611、0.4884、0.4750和0.7500,平均杂合子比例为0.4829。研究结果显示,栉孔扇贝有丝分裂雌核发育二倍体均为纯合子,如精子的灭活率达到100%,有丝分裂雌核发育二倍体一代即可实现纯合,如再进行一次减数雌核发育即可建立纯系;减数分裂雌核发育二倍体由于具有较高的重组率,其与母本的遗传同质性较高。 We used the genotyping data of five microsatellite loci to check the absence of any paternal contribution to the gynogenetic offspring genome, to estimate the M-C recombination rate and to analyze the homozygous ratio in the induced gynogenetic diploids. In this study, gynogenetic diploid Chlamysfarreri were induced by inhibiting exclusion of the second polar body and by suppressing the first cleavage with 6-DMAP in eggs fertilized with UV irradiated sperm. Five SSR heterozygous loci in Chlamysfarreri were used to identify the homozygosity of 30 larvae in the gynogenetic families A and B, 40 larvae in their control groups and their parents. Electrophoretic patterns showed that all the 5 pairs of microsatellite primers produced well-identifiable DNA fragments. The results of the five microsatellite analyses revealed that the presence of male DNA in the two gynogenetic families indicated they were derived from normal fertilization. 6-DMAP treatment produced 68.5 % gynogenetic diploids. All homozygosity in Mitotic-A and Mitotic-B had a high percentage of heterozygosity in Meiosis-A and Meiosis-B. The rate of heterozygosity in meiogynogenetic progenies at the loci of CFMSPO07, CFMSP075, CFMSMO09, CFMSMO14 and CFMSM020 were 0.340 0, 0.361 1, 0.488 4, 0.475 0 and 0.750 0, respectively. The average heterozygous rate of the 5 loci was 0.482 9. The high proportion of heterozygosity for the 5 loci demonstrates that it is not a practical method for producing homozygous inbred lines in the gynogenetic scallop produced by retention of the second polar body, and treatments of suppressing the first cell division were more promising for this purpose. In addition, microsatellite analysis is an effective method for the verification ofgynogenesis because of their codominant and highly polymorphic nature. [Journal of Fishery Sciences of China, 2008, 15 (1) : 55-62]
出处 《中国水产科学》 CAS CSCD 北大核心 2008年第1期55-62,共8页 Journal of Fishery Sciences of China
基金 国家自然科学基金资助项目(30600465) 国家科技攻关项目(2004BA526B0103).
关键词 栉孔扇贝 雌核发育 微卫星 纯合性 Chlamysfarreri gynogenesis microsatellite homozygosity
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