长牡蛎SNP标记开发及其在家系分析和物种鉴定中的应用.doc

长牡蛎 SNP 标记开发及其在家系分析和物种鉴定中的应用 Development and characterization of SNP markers and their application in family analysis and species identification for Pacific oyster Crassostrea gigas Abstract Crassostrea gigas (Pacific oyster) belongs to mollusks, widely distributed along the seashore of China, Korea, and Japan, is a bivalve with wide tolerance of temperature and salinity. It is the top-drawer species in the world shellfish industry, and become one of the most important cultured species in China. Owning to the characteristics of high-density genome-wide distribution, genetic stability, co-dominance, and allowing high-throughput assays, single nucleotide polymorphisms (SNP) is a kind of ideal molecular marker for the genomics and genetics research. Now, redundant sequences found in EST database can be a useful way for mining SNP markers or other mutation sites. High resolution melting curve (HRM) has been developed to detect and genotype SNPs because it is a simple, rapid and cost-effective technology. This research is to develop and characterize SNP markers of C. gigas by HRM method, and apply SNPs in family analysis and Crassostrea species identification. The results provide new insight of SNPs in the fields of high-density genetic mapping，biodiversity conservation，population structure analysis and marker-assisted breeding. The development and characterization of SNPs Though mining EST database and manual quality filtering, a total of 48769 putative SNPs were detected from 46171 ESTs. 55 gene-derived SNPs were isolated and characterized by means of HRM analysis. The observed and expected heterozygosities ranged from 0.063 to 0.563 and 0.091 to 0.448, with an average of and 0.279, respectively. The PIC ranged from 0.059 to 0.371 with an average of The combined power of exclusion was greater than 99.99% across all these SNPs. The application of SNPs in family analysis All the 55 polymorphic SNPs were analyzed in six C. gigas single-pair mating families. No null al