The Study On The Isozymic Tissue Specificity Of Two Native-born Fish In Yangtze River And The Isolation Of Microsatellites In Genomic Of Spinibarbus Sinensis (Bleeker)

Spinibarbus sinensis (Bleeker) and Procypris rabaudi (Tchang) are famous and precious economical native-born fish in China’s Yangtze River basin, they are important culture fishes in Chinese aquaculture. For protecting the germplasm resource of Spinibarbus sinensis (Bleeker) and Procypris rabaudi (Tchang), two aspects covered by our study are the isozymes and the microsatellite DNA makers in them.At first, by using the vertical polyacrylamide gel electrophoresis, Lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and esterase (EST) isozymes from six tissues (brain, eye, heart, liver, kidney and muscle) of adult Spinibarbus sinensis (Bleeker) and Procypris rabaudi (Tchang) were preliminarily studied. The loci and phenotypes of the isozymes in every tissue were also analyzed. The results showed that all isozymes of them presented tissue specificity. The result of Spinibarbus sinensis (Bleeker) showed that five classical LDH bands were found in the tissues, which was encoded by three loci, and the site C was only found in liver. The number of MDH isozyme bands was three in five tissues except muscle, which were all s-MDH. Both s-MDH and m-MDH were found in muscle. EST isozyme was encoded by five loci at least. The isozyme pattern was complicated, and the stronger activity was pigmented in the liver and kidney. The result of Procypris rabaudi (Tchang) showed that LDH was encoded by three loci, and the site C was only found in liver; s-MDHs had been found in six tissues of Procypris rabaudi, and the stronger activity was pigmented in the liver and kidney. There were m-MDHs both in four tissues(brain, heart, kidney and muscle); EST isozyme was encoded by five loci, and the isozyme pattern was complicated. Est-2was the common band in six tissues, and the stronger activity was pigmented in the liver.Twenty-four pairs of microsatellite primers developed from Procypris rabaudi were used to scan the genomic DNA of Spinibarbus sinensis (Bleeker), for the analysis of the polymorphism microsatellite DNA makers in Spinibarbus sinensis (Bleeker). Througth the molecular techniques such as the extraction of genomic DNA、PCR amplification and gel electrophoresis and so on,10pairs of primers were screened effective and4microsatellite loci showed polymorphism in Spinibarbus sinensis (Bleeker). The number of alleles per locus ranged from6to10, the size is between lllbp-258bp, the ranges of observed heterozygosity (Ho) were0.750-0.833, average observed heterozygosity was0.771. The ranges of expected heterozygosity (He) were0.688-0.837, average expected heterozygosity was0.786. The average PIC was0.726. All polymorphism loci were in Hardy-Weinberg equilibrium (HWE).