| Trypsin (TRY) and Amylase (AMY) are critical enzymes in the dietary protein and sugarlevels, digestibility and metabolism. In this study, single nucleotide polymorphisms (SNPs) ofTRY gene and AMY gene and SSR of AMY gene were detected in Siniperca chuatsi by directsequencing (DS). We found a novel SNP site (G169A) within the exon3in TRY gene and fourSNPs in AMY gene. Two hundred Siniperca chuatsi were tested with DS method and the resultsillustrated three unique binding genotypes of GG，GA and AA. Frequencies of the allele G and Awere0.53and0.47. Created restriction site (CRS), single stranded conformation polymorphism(SSCP) and single tube bidirectional allele specific amplification (SB-ASA) methods were usedto identify the SNP site. The results showed that the method of CRS-PCR could not test this SNPsite, both SSCP and SB-ASA techniques could identify this site and the accuracy of SSCP was100%and that of SB-ASA was96.67%, respectively. PCR-direct sequencing (PCR-DS) methodwas used to locate the SNPs and SSRs, and SNPs genotyping was carried out with createdrestriction sites PCR (CRS-PCR)method in AMY gene. In addition, polyacrylamide gelelectrophoresis was used to check the polymorphism of SSR.In our study we found that there areone SNP A1in intron1of AMY gene, and three SNPs (A2, A3and A4) locate in a SSR ofintron5. This SSR locus B1has4alleles and9genotypes in total. Chi-square test was carriedout to examine the G169A, SNP-A1and SSR-B1, results indicated that the genotypes of G169Alocus has no significant difference between the domesticated and undomesticated populations ofSiniperca chuatsi(P＞0.05), neither the genotypes of SNP-A1and B1locus has(P＞0.05).These SNPs and SSR can consider as genetic markers for feed habit domesticationphenotype in Siniperca chuatsi. In addition, this study laid the foundation for developement offeeding domesticated cultivation of Siniperca chuatsi research in future. |
PDF Full Text Request