异源精子诱导栉孔扇贝雌核发育后代的微卫星分析
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国家“八六三”高技术研究发展计划(2006AA10A408);国家科技支撑计划专题(2006BAD01A00);国家自然科学基金资助(30600465)


Microsatellite analysis of gynogenetic diploids induced by heterologous sperm in Chlamys farreri
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    摘要:

    采用紫外线遗传灭活的长牡蛎精子激活栉孔扇贝卵子,6-DMAP诱导染色体加倍的方法,获得第二极体抑制型雌核发育二倍体早期胚胎。通过优化PCR反应条件,在11对栉孔扇贝和10对长牡蛎微卫星引物中共筛选出3对引物,可以同时在长牡蛎和栉孔扇贝基因组中获得稳定性,多态性较好的特异性扩增条带,其中2对引物为栉孔扇贝引物。利用筛选出的3对通用微卫星引物对雌核发育后代进行检验及遗传分析。结果表明,雌核发育个体基因完全来自于母本,后代中没有父本基因的表达;部分雌核发育后代在3个座位上发生了纯合,部分个体发生了不同程度的基因重组,3个座位上的重组率分别为40%、55%和35%。研究结果从分子水平上证实,利用遗传失活的长牡蛎精子诱导栉孔扇贝雌核发育是可行的,只进行一次第二极体抑制型雌核发育二倍体的诱导只能获得部分后代个体的基因纯合,但后代与母本具有较高的遗传同质性。

    Abstract:

    The Zhikong scallop Chlamys farreri plays an important role in the shellfish breeding in the north of China and the gynogenesis in this scallop has developed rapidly in recent years. In this study, the meiotic division of the eggs of C. farreri was activated by ultraviolet (UV)irradiated heterogenous sperms of Pacific oyster Crassostrea gigas and the gynogenetic diploids were induced by blocking extrusion of the second polar body from fertilized eggs with 6dimethylaminopurine (6-DMAP). To assess homogeneity and isogenedity between the progeny and their parents, microsatellite technique were used. Then 3 pairs of common microsatellite primers to be used for genetic analysis of offspring were screened from 21 pairs of microsatellite primers among which 11 pairs were indentified in C. farreri and 10 pairs in C. gigas by optimizing the condition of polymerase chain reaction(PCR). Under the optimal condition, PCR was performed for analyzing the genetic character of the gynogenetic offspring. It was found that the 3 loci could amplify clear fragments in these offspring. The results of genetic analysis showed that the gene of offspring without gene contamination of male parent C. gigas was completely inherited from female parent C. farreri. Microsatellite locus and centromere recombined at the 3 polymorphic loci and the recombination rates were 40%, 55%, and 35%, respectively. This study confirmed that it was feasible to achieve diploid gynogenetic progeny of C. farreri by heterologous sperm’s inducement while it was difficult to get an absolute homozygotic population once in manner of suppression of the meiosisⅡ, because there was high heterozygosity in the first meiogynogenetic generation owing to crossover between the homologous chromosome during the meiosisI.

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吴彪,杨爱国,王清印,刘志鸿,周丽靑.异源精子诱导栉孔扇贝雌核发育后代的微卫星分析[J].水产学报,2009,33(4):542~548

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  • 收稿日期:2008-05-30
  • 最后修改日期:2008-09-22
  • 录用日期:2009-06-15
  • 在线发布日期: 2009-07-04
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