Genetic Diversity Studies Of Pinus Sylvestris Var.Mongolica Litv Under Different Ceographical Environments

In order to study the genetic diversity of Pinus sylvestris var. mongolica Litv. in different environments, propose a theoretical basis for artificial introduction promotion in the future, and explore introduced recession during introduction, four different locations Pinus sylvestris populations (such as Weichang population, Honghuaerji population, Yulin population and Zhanggutai population.) were selected to study genetic diversity. In this study, Pinus sylvestris var. mongolica Litv. polymorphic EST-SSR primers were developed based on Pinus tabuliformis transcriptome data, and Pinus sylvestris SSR optimum reaction system was established by optimizing the reaction system. It was the first time that EST-SSR marker was used to analyse Pinus sylvestris genetic diversity comprehensively in different population. The genetic diversity inter or inner population was clarified. While the correlation analysis in different environmental factors illustrated relationship between genetic variation and ecological factors. This study proposed a theoretical basis to artificial introduction promotion, and provided scientific guidances to genetic resources conservation.The results showed that:1. SSR reaction system optimization results has been got. At first, Pinus sylvestris var. mongolica Litv. in Weichang population was selected as simples. Univariate multi-level experiment was carried on for4factors (Template DNA concentration, Taq DNA polymerase content, dNTP concentration, primer concentration), and then the preliminary results of the optimization came out Second, orthogonal test L16(45) for4factors in4levels was conducted to optimize the result. At last, the SSR reaction system optimization results for Pinus sylvestris var. mongolica Litv. was carried out.2.25pairs of primers was screened from175pairs which are polymorphic in Pinus sylvestris var. mongolica Litv.. Those SSR loci are dinucleotide repeats (8,32%), trinucleotide repeats (11,44%) and other repeat types are six, accounting for24%. After BlastX and Gene Ontology analysis,12potential function sites were found, accounting for48%of the total.3. Genetic distance between four population and factors affecting genetic diversity has been reached. Four population cluster analysis using15polymorphic primer pairs showed that genetic distance is closer in Zhanggutai and Honghuaerji, followed by the Weichang, the farthest is Yulin. Multiple regression analysis showed that the genetic diversity may be affected by geographic isolation.4. According to research data, protection and the introduction suggestions have been put foword. In four groups, only Yulin is in line with Hardy-Weinberg equilibrium, and the most linkage disequilibrium loci emerged in Zhanggutai population, may the seed orchards in this region suffered excessive human intervention. In the process of genetic resouces protection, a recession cased by frequent inbreeding should be avoid. Artificial introduction could be extended to similar environment or close region.