Genetic Diversityanalysis Of A Rare And Protected Bamboo Of Qiongzhuea Tumidinoda

Qiongzhuea tumidinoda was a rare and Protected Bamboo Species in China, studied of genetic diversity of species for the conservation and utilization was importance. Microsatellite(SSR) and AFLP molecular markers technique were applied for the genetic and diversity study of two bamboo populations, which collected from Sichuan xuyong and Yunnan Suijiang . The successful using of two molecular markers provided a theoretical basis and technical support for the conservation and utilization of Qiongzhuea. The main results were as follows:1. Through the comparison and the improvement of plant genome DNA extraction method, a quite suitable Qiongzhuea tumidinoda DNA extraction can be obtained. High-quality DNA which is suitable to molecular marker technique can be extracted from bamboo blade.2.10 polymorphic primers, which amplified clear bands with high SSR polymorphism, were screened from that were broadly used in other tree species. Effects of Taq polymerase, concentrations of template, Mg2+, dNTP and primers on the results of SSR-PCR were analyzed. A stable and repeatable reaction system for SSR-PCR and its parameters were constructed. The results showed that the optimum quantities of template, Mg2+, dNTP, primer and Taq polymerase were 60ng, 1.5mmol·L-1, 0.2mmol·L-1, 0.25μmol·L-1 and 0.8U in the 20μL system, respectively.3. Genetic diversity of Qiongzhuea was analyzed by SSR with 10 primer combinations. The Population of Qiongzhuea showed a high genetic diversity, whose Nei's genetic diversity was 0.7874. The mean observed heterozygosity was 0.7359, more then that of expected heterozygosity(He,0.7315). There was no significance between observed heterozygosity and expected heterozygosity except for RM245.F was -0.0060,close to 0, indicting that no significant diference existed between heterozygosity and homozygotes of the parentage population. The value of Fst ranged from 0.0123(RM205)to 0.0938(RM337)and averaged out 0.0459,that showed a 4.59% of genetic variation between populations, indicating that genetic variation within population was the primary(95.41%). The Nm value was 5.2004, larger than 1, indicating that genetic difference among populations of Qiongzhuea was not resulted from genetic drift.4. Ten EcoRI/MseI primer pairs showing high level of polymorphism and distinct band were selected out and used for further practical AFLP analysis. 680 bands were amplified, each primers can amplify68 bands and 663 of 680 amplified bands were polymorphic bands, which accounts for 97.20%.5. The rate of polymorphism were89.86% and 91.95%; the number of alleles(na) were 1.8986 and 1.9195; the effective number of alleles(ne) were 1.5850 and 1.5683; the average Genetic identities within 6 populations(h) was 0.3321; the average Genetic identities with in 2 populations(h) was 0.3949; Shannon information index(I) was 0.5754.the genetic differentiation coefficient among 2 populations(Gst) was 0.1488.The gene flow among 2 populations(N) was 2.8610.The average genetic identity was 0.8219 and the average genetic distance was 0.1961.The 2 populations were divided according to the UPGMA cluster analysis.