| Half smooth tongue sole (Cynoglossus semilaevis) females grow bigger and mature later than males, it was reported that females grow2-3times faster than males. So the production of all-female stock would be of significant benefit for aquaculture of half smooth tongue sole.Here an effective method to induce gynogenesis for all female population construction in half smooth tongue sole was provided and the cytological processes of induced meio-and mitogynogenesis were studied. It also can study the sex determination mechanism. MAS is useful for assisting and speeding up selection through conventional breeding.Gynogenesis was induced in half smooth tongue sole by using UV irradiated cryopreservation of sea perch sperm, the gynogenesis was induced by the cold shock and hydrostatic pressure. Detect the sex type and radio in the offspring of meiogynogenesis and mitogynogenesis. Designed the primers depend on linkage map, and correlation analysis between growth-related traits and microsatellite markers in half smooth tongue sole.1. By the cold shock and hydrostatic pressure, gynogenesis was induced successfully. In the method of cold shock, the best parameter is DA(treatment after fertilization)5min,25min of D (shock duration), T(shock temperature) was5℃, put the embryo into sea water (24℃) after the test. In the hydrostatic pressure, DA was21.5min, D4min, the shock pressure was60MPa.2. Detected the sex type and radio in the offspring of gynogenesis with Cyse-SSRl. In meiogynogenetic diploids six WW "super-female",34ZZ male gynogens were observed in40embryos (blastula stage). No WW "super-female" was observed in40embryos (Sarcomere period, newly hatched larva), they were all ZZ male. In mitogynogenetic diploids five WW "super-female",35ZZ male gynogens were observed in40embryos (blastula stage). No WW "super-female" was observed in40embryos (Sarcomere period, newly hatched larva), they were all ZZ male. In control no WW "super-female" was observed, ZW:ZZ=1:1.3. Detected the sex type and radio in the offspring of gynogenesis with chromosome technology. In gynogenesis group no WW "super-female" and ZW female was observed. All of them were ZZ male. In control ZW:ZZ=1:1.4.84microsatellite loci were selected to amplify the genomic DNA of cultured stock of half smooth tongue sole. Genotypic differences of these microsatellites loci were analyzed using%2test between the maximal weight group and minimal weight group. Twelve microsatellites showing significantly difference (P<0.05) were used in genotyping individuals. There were csou34, ghrh-ssr, hncyse160(P<0.01). cs7, csou4, cyse22, cyse72, cyse123, sox9-1, hncse67, hncyse124, hncyse129(P<0.05)。5. Divided the random population into two group, one is female group(45individual), the other one is male group (51individual). The associations between their genotypes and growth traits were examined. The results suggested that csou4, cyse123, ghrh-ssr, cyse72and hncyse160in female; hncyse129, hncyse160, sox9-1in male were associated with body length. Ghrh-ssr in female; csou34, cyse22, cysel23, hncse67, sox9-1in male was associated with body depth. Csou4, ghrh-ssr, cyse72, hncyse160in female was associated with body weight. No microsatellite loci was associated with body weight6. The result of multiple comparison showed that in female group AA of cs7, AE of csou4, AA of csou34, EF of cyse22, BC of cyse72, AB of cyse123, BC of hncse67, AC of hncyse124, CC of hncyse129, BB of hncyse160, AE of ghrh-ssr, AD of sox9-1were superiority genotypes. In male group AA of cs7, AC of csou4, EE of csou34, CD of cyse22, BD of cyse72, AA of cyse123, AC of hncse67, BC of hncyse124, CF of hncyse129, BB of hncyse160, AC of ghrh-ssr, AB of sox9-1were superiority genotypes.7. Genotype AE of csou4, BC of cyse72, BB of hncyse160and genotype AE of ghrh-ssr might be regarded as the candidate market for body weight trait. The results also showed that the more body associated markers, the larger body weight... |
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