Isolation Of Microsatellite Loci From Haliotis Diversicolor And Pseudosciaena Crocea

Using the FIASCO(Fast Isolation by AFLP of Sequences COntaining repeats) method, we constructed the (AC)n enriched library for Pseudosciaena crocea and the (CT)n enriched library for Haliotis diversicolor, respectively. We obtained many microsatellite sequences from the libraries, while some new markers were identified.A total of 1212 colonies was contained in the (CT)n enriched library for H. diversicolor. One hundred and fourteen colonies were sequenced and 48 positive colonies containing 10 or more (CT) repeats were obtained, in which 26 sequences were fit to design the primers for amplifying the microsatellite loci. But only 19 loci were successfully amplified and 11 loci were polymorphic. Then, 24 wild individuals from Hainan province were used to test the 11 polymorphic loci. These polymorphic microsatellites showed allele numbers ranging from 3 to 10, and PIC (polymorphism information content) ranging from 0.148 to 0.818. The observed heterozygosities (Ho) and expected heterozygosities (He) ranged from 0.083 to 0.913 and from 0.159 to 0.856, respectively. All loci were in Hardy-Weinberg equilibrium (HWE) except SA-JMU4 and SA-JMU12(P<0.01).The (AC)n enriched library for P. crocea contained 1440 colonies. Of 257 colonies sequenced, 182(70.8%) were positive for a microsatellite insert (n≧10). We tested 10 loci from 26 primer pairs for their polymorphism. The number of alleles ranged from 3 to 12, and PIC ranged from 0.283 to 0.866. The observed heterozygosities (Ho) and expected heterozygosities (He) ranged from 0.233 to 0.900 and from 0.326 to 0.893, respectively. The loci showed no evidence of linkage disequilibrium. Six loci departed significantly from HWE(P < 0.05), except LYC0044,LYC0046,LYC0053 and LYC0060.In this paper, the construction of microsatellite enriched library and the improvement of efficiency for enriched library were discussed. And also the genetic variability of wild population for P. crocea and H. diversicolor were compared with the results reported by other researchers. These novel markers isolated in this study are useful to all the field since most of their PIC are higher than 0.5.They will be helpful to researches for H. diversicolor and P. crocea.