Development Of Single Nucleotide Polymorphism Markers In Growth And Biomineralization Genes Of Pearl Oyster Pinctada Martensii

Pearl oyster Pinctada martensii (Dunker) is the main species for marine pearl production in China.Pearl oyster growth traits (such as shell height, shell width and shell thickness, shell weight and total weight) and the secretion of nacre traits (such as nacre thickness, pearl quality, etc.) is the primary goal of genetic improvement of Pinctada martensii. However Screening with the growth and nacre secretion traits are significantly associated molecular markers for assisted selection, as Pinctada martensii growth and fertility beads traits for effective choice of efficient and reliable means and Approaches. Integrating informatics strategies " is a method of screening analysis of candidate genes in animal breeding of poultry and other agricultural, that systematic analysis Pinctada martensii ESTs of we establishment and gained. In this study, screening significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Use of amplified re-sequencing Technology to detect and Development single nucleotide polymorphism (SNP) loci. Detection of genetic characteristics and genetic model in the wild populations and family of Pinctada martensii, and then Detection of SNP markers associated with the growth and nacre secretion traits in the Segregating populations. In addition, due to the advantages of Richness in the genome distribution and the same Sites Biallelic make more refined in genetic mapping. SNP-based linkage map not only increased the marker density, provides the physical map with the rich genetic link, thus contributing to the integration of genetic maps and physical maps; and provides a detailed map of quantitative trait loci and QTL gene identification.In this study, bioinformatics analysis of the EST database of Pinctada martensii, get the significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Primers were designed for174, Amplification and amplification products re-sequenced in12individuals of six different geographical populations in Pinctada martensii. Use of Align analysis of the measured, three hundred and fifty-four putative SNPs were discovered by resequencing33Kb, corresponding to a very high density of one SNP per94bp. Select the40SNPs in all sites, Primers were designed for each SNPs,27pairs of primers showed a single band on polyacrylamide gel. Design length of20-35bp probe for the27SNPs, After PCR, the non-labeled probe High Resolution Melting (HRM) to detect the SNP genotype. In three wild populations(popl:Collected from Sanya Yalong Bay in2003、pop2: Collected from Sanya anyou in2009pop3:Collected from Sanya Nanshan in2010) and a family (48individuals) of Pinctada martensii, the17SNP markers to obtain a stable genotype data. The one marked as a single state, the remaining16markers were polymorphic. of which act showed significant (P <0.05after Bonferroni correction) departures from Hardy-einberg equilibrium in all the populations with heterozygote excess,while pris-2,mec-7a and hsp-1deviated from HWE in at least one population. act and mec-7a in the deviation from the group showed a significantly to heterozygotes excess (P <0.01), act and pris-2in the deviation from the group showed a significant to deficiency (P <0.01). These sites may be subject to selection pressure.16SNPs can be amplified in the vast majority of individuals in three wild populations, the overall average Fst was only0.0048. Description of the16genetic differences was not significant between the three groupsFourteen (excluding mec-7b and ben-1) of the16validated SNPs segregated in a mapping family, ddx-23and hsp-1significantly deviated from the Mendelian ratios(P<0.05), Show these two sites subject to selection pressure. View of all SNP loci derived from ESTs sequences that related to the growth of gene and shellfish biomineralization of nacre protein gene, these sites Related Traits in analysis and position on the genetic linkage map and QTL analysis will reveal where the gene function and its mode of action. Lay the foundation for the Pinctada martensii molecular marker assisted breeding and functional genes.