Effective Population Of Content Group Of Core Collection Of Portunus Trituberculatus

This article based on the Portunus trituberculatus core group of fast-growing strain of "Huangxuan NO.1"as the experimental materials.According to the formulas of effective population size ,which brought forward by Doyle and Talbot,we estimated the effective population size(Ne)for rapid growth line fundamental stock in P.trituberculatus. In 2009, we selected the fast-growing strains of the offsprings through winter, according to the effective population Ne = 1,2,3,5,10,20,50 gradient production, compare the content of their offspring effective population.Selecting 22 pairs of polymorphic microsatellite markers on P. trituberculatus, building multiple PCR amplification technology system after matches between the primers and combinatorial testing. three four PCR combinations were also established. In this study, multiplex PCR reaction conditions were optimized, including annealing temperature, Mg²⁺ concentration, dNTPs concentration and other parameters.Finally,15 combinations of triple PCR were established, three four PCR combinations were also established.Using one of the two groups of triple combination, a quadruple combination of core infrastructure group P. trituberculatu.rapid growth of strains 2008 and 2009 were paternity test,the results are as follows:According to the formulas of effective population size ,which brought forward by Doyle and Talbot,we estimated the effective population size (Ne) for rapid growth line fundamental stock in P. trituberculatus.The real Ne was 213, which corresponded to a rate of inbreeding (ΔF) of 0.23%.Results indicated thatΔF is relatively small,and inbreeding depression was in a low level, however, the cost of conservation was too large, it did not meet the conservation requirements. In light of breed conservation model in livestock and poultry, we investigated rapid growth line fundamental stock in P. trituberculatus on methods of selecting parents and ratio of dams and sires and the size of conservation population. We make the rules as follow: random selecting parents, the ratio of dams and sires is 1:1, and the number of dams and sires is 131, respectively. In this situation, genetic diversity of rapid growth line fundamental stock is preferably rich,which satisfies selective breeding, at the same time the cost on breed conservation is economical. This is only a theoretical model of breed conservation, and it needs the process of gradual improvement of breeding populations in the future. In 2009, we selected the fast-growing strains of portunus offspring crab through winter, press the effective population Ne = 1,2,5,10,20,50 gradient production, its Daphnia IV and megalopaâ…¡crab survival rate of the measurement period, the survival rate is not significantly different (P <0.05), after the larvae into an outdoor culture, 80 days, 100 days, 120 days to measure their growthfind that the different effective population size of the growth trituberculatus differences (P> 0.05).Increase the effective population size of the growth and survival of F₁ no significant effect, it is worth noting that although in the short term this effect (F₁) aspects of the phenotype does not exist, but because of some small group of individuals to participate in reproduction, and itsgenerations of individuals within the group closer genetic relationship, the lower genetic diversity, and thus long-term impact on the (F2 and beyond) can not be ignored, which will be explored in future research.Selecting 22 pairs of polymorphic microsatellite markers on P.trituberculatus, building multiple PCR amplification technology system after matches between the primers and combinatorial testing. 47 combinations of double PCR were established, 15 combinations of triple PCR were established based on the 47 double PCR combinations, three four PCR combinations were also established. In this study, multiplex PCR reaction conditions were optimized, including annealing temperature, Mg²⁺ concentration, dNTPs concentration and other parameters. Multiple PCR technology system was established eventually, the establishment of this technology provides a fast, effective molecular detection technology for the portunus breeding, germ-line assessment.Selecting polymorphic PCR products with few null allele sites, the final combination of select 2 tripl notyping and Cervus3.0 artificial simulation method for its identification rate of 100%, so it can quickly understand the various families of the survial in"Huangxuan No. 1", guiding the breeding of"Huangxuan No. 1". According to each parent's contribution of the different generations, as well as the number of parents, the calculated size of inbreeding coefficient incrementΔF= 0.0058, effective population size Ne = 88, small increments of inbreeding coefficient, remained low level, building the family level more in line with group selection. In summary, microsatellite multiplex PCR gene scanning technology supplies a fast and efficient tool for accelerating the breeding process of P. trituberculatu.