大黄鱼与黄姑鱼异源三倍体的诱导和微卫星分析  

Induction and micosatellite analysis of allotriploid between Larimichthys crocea and Nibea albiflora

作　　者：蔡明夷[1] 刘贤德[1] 武祥伟[1] 黄铭垚[1] 王志勇[1] 

机构地区：[1]集美大学水产学院,福建省高校水产科学技术与食品安全重点实验室,福建厦门361021

出　　处：《水产学报》2010年第11期1629-1635,共7页Journal of Fisheries of China

基　　金：国家“八六三”高技术研究发展计划(2006AA10A405);集美大学创新团队基金(2006A001);中国科学院海洋生物资源可持续利用重点实验室开放基金(LMB091007)

摘　　要：参照大黄鱼雌核发育诱导程序,应用冷休克抑制大黄鱼(♀)与黄姑鱼(♂)杂交受精卵的第二极体排出,培育了两个异源三倍体家系(PPN1和PPN2)。异源三倍体家系的受精率、孵化率略低于大黄鱼自繁二倍体对照家系(PP),而初孵仔鱼畸形率略高于PP家系。倍性分析显示,异源三倍体家系初孵仔鱼细胞DNA含量约为大黄鱼自繁对照家系的初孵仔鱼细胞DNA含量的1.46倍,且三倍体率达到100%。5个微卫星标记分析结果表明,父本杂合基因在异源三倍体中分离,后代分别得到父本两个等位基因中的一个,分离比符合孟德尔式遗传预期;由于基因的第二次分离被阻断,母本基因在异源三倍体中的传递表现出半四分子的特点,其中部分个体同时保留了母本两个等位基因,表现为杂合基因型。综合倍性分析和微卫星分析结果可以判断,异源三倍体家系成员为含有2个大黄鱼基因组和1个黄姑鱼基因组的异源三倍体。可见,大黄鱼减数分裂雌核发育或三倍体诱导程序中用于抑制第二极体排放的条件,同样适用于诱导异源三倍体。然而,PPN1和PPN2的仔鱼在15日龄后出现生长停滞,陆续死亡,没有个体存活超过1个月,表明母本染色体加倍不能有效提高杂种成活率,但异源三倍体仔鱼可作为遗传作图、基因组比较和基因表达调控研究的材料。The developed hybrid eggs of Larimichthys crocea（♀） and Nibea albiflora（♂） were shocked with cold treatment to block the extrusion of the second polar,according to the protocol of inducing meio-gynogenesis in large yellow croaker.Two allotriploid families（PPN1 and PPN2） and the control families were produced.The embryos developed normally,and the morphologic normal fry were hatched out.The development rate,hatching rate and normality rate of the allotriploids were slightly lower than those of the diploid control of large yellow croaker.The results of ploidy analysis showed that the DNA content of the newly hatched fry in PPN1 family was 1.46 time that of diploid large yellow croaker,and the 100% tested fry in PPN1 were triploid.Each offspring in allotriploid families inherited one of alleles from its male parent.Since the gene segregation in Meiosis II was blocked,both homozygotes and heterozytos were found in allotriploid families at four maternal heterozygotic microsatellite loci.The results of the ploid analysis and microsatellite analysis proved the putative triploids contained two genomes from large yellow croaker and a genome from yellow drum.However,the allotriploid fry died one by one from 15 th day post hatching and none survived over one month old,showing that the diploidization of the maternal genome did not improve the survival of the hybrids.Nevertheless,the allotriploid fry can still serve as good materials in genetic mapping and to reveal gene expression and regulation in some important biological processes.

关 键 词：大黄鱼 黄姑鱼 异源三倍体 倍性分析 微卫星 

分 类 号：S917[农业科学—水产科学]