通过精液移植构建中国明对虾家系及微卫星分型鉴定父本

Semen transplantation and paternity assignment by microsatelliteg enotyping in Fenneropenaeus chinensis

本研究首次采用精液移植法进行中国明对虾(Fenneropenaeus chinensis)的家系构建,利用10尾已交尾雌虾纳精囊内精液对17尾未交尾雌虾进行了移植,共建立11个中国明对虾家系,其中含有父系半同胞家系2个;精液移植的平均受精率为46.8%,最高受精率为78%,授精成功率为64.7%。为了揭示每个家系母本内精液的来源情况,选用6个中国明对虾微卫星标记对11个家系的母本、母本内精液及每个家系的6个个体进行分析,推算出每个家系各自的父本基因型,并与各自母本内精液基因型进行比较,成功地对11个家系进行了父本鉴定。实验结果表明,11个家系的父本基因型与各自母本内精液基因型相同,每个家系内个体属全同胞关系,即对于1尾自然交尾雌虾,其纳精囊内精液只来自于1尾雄虾。通过本实验,验证了精液移植在构建中国明对虾全同胞家系和半同胞家系中的可行性,阐明了精液移植家系内个体的亲缘关系,为中国明对虾选育工作提供了技术保证。

One of the key factors in the process of large-scale families breeding of Fenneropenaeus chinensis is the construction of base population with broad genetic variation.The base population can be established by forming a synthetic breeding population where brood stock from different populations are systematically crossed with each other to secure that their offsprings do not inbreed but carry new combinations of alleles.Howerer,lack of male brood stocks restricts the establishment of enough families.In order to resolve this problem,we used semen transplantation to construct families of F.chinensis among different populations,and for the sake of obtaining pedigree information, microsatellites were used for identifying parentage relationships among individuals. Semen from spermathecas of ten mated shrimps was transferred into the spermathecas of 17 shrimps which were not mated,and 11 full-sib families were obtained,including two half-sib families.The average fertilization rate was 46.8%with the highest being 78%.The success rate of artificial insemination by semen transplantation was 64.7%. Six microsatellite markers were used to identify parentage relationships among the individuals.We identified paternity of the 11 families through genotype data of female parents,semen and six individuals of each family,and constructed clear pedigrees by using microsatellite loci.All of the six microsatellite loci were polymorphic.Forty alleles were detected in 11 female parents at six microsatellite loci.The numbers of alleles at each locus were between four and nine.Forty-two kinds of genotypes were acquired in 11 female parents.All the paternal genotypes of the 11 families were inferred based on the genotypes of the female parents and their offsprings.Through the analysis of expected sire genotypes and semen genotypes,we successfully identified the paternity of 11 families.Results indicated that the male parent genotypes were identical with the semen genotypes,which demonstrated that semen in each spermatheca was from one male shrimp,and the offsprings of each family were full-sibs.These results demonstrate that it is feasible to use semen transplantation technique as a way to establish F.chinensis families.It also shows that microsatellite markers can be used as a powerful tool for obtaining pedigree information in selective breeding programs of F. chinensis.The results of the experiment will establish foundation for producing full-and half-sibs families,and the technology of semen transplantation can provide basis for shrimp breeding programs.