Analysis On The Genetic Diversity Of Persimmon By SCoT Makers

Persimmon(D/ospyros kaki Thunb.) is a species of Diospyros L. in Ebenaceae on botanical classification, which has about3000years of culture history in China. The persimmon resources in China are very rich, the varieties are over one thousand. These plenty of materials play an important role in the identification, genetic diversity analysis, genetic improvement and breeding. For a long time, people still lack of comprehensive understanding about persimmon germplasm resources, and the background of most Chinese persimmon germplasm was not clear, and research on the origins of the Diospyros kaki, evolution and relationship still lack in-depth discussion.97different types of persimmon were selected from National Field Genebank for Persimmon as matierals to study through morphological traits and SCoT moleculer markers, which including24Niuxinshi and22Yeshi;24bud mutant varieties from Fuyuu, Jiro, Gosho, Jianshi, the hybrid varieties bred in Japan;17similar persimmon with different names;10hybrid combinations and their offspring. In this study, we use morphologic observation and SCoT markers to analysis97Diospyros kaki materials for their genetic diversity, in order to research and analysis the phylogenetic relationships among species and varieties from the DNA level. The main results are as follows:1. In this experiment, a L16(45) orthogonal design was used for the optimal SCoT-PCR reaction system. The optimal PCR reaction mixture of SCoT-PCR system was2.0mmol/L Mg2+,0.2mmol/L dNTPs,1.0U Taq polymerase,0.5μmol/L primer and80ng template DNA in20μL mixtures.2.10primers were selected from40ones of SCoT in97different materials, which could produce stable and polymorphic bands; Furthermore, the annealing temperature of each selected primer was optimized, the control temperature range from45.0℃to55.0℃, found that different primer has different annealing temperature, which can increase the specificity of the amplified product.3. By NTSYS-pc2.1software we had the cluster analysis sorts with the SCoT data, obtained the cluster figure of persimmon germplasm resources, analysised the genetic relationships among species, and compared field analysis. 4. Yemaoshi with staminate red flower and pistillate red flower were gathered for a group, they have far relationships with Niuxinshi group and Yeshi group, which is consistent with the morphologic observation, can be considered as a new species. Bishan niushishi has far relationships with other Niuxinshi varieties, which is close to the original wild traits in morphological, Bishan niushishi is tetraploid, different from others which are hexaploid, Bishan niushishi should belong to Yeshi, so it is reasonable to be clustered on Yeshi group.24bud mutant varieties were divided into five groups, each group has difference in degrees, basically consistent with the phenotype observed.17similar species with different names are largely clustered together, which is consistent with the morphologic observation. In hybrids combinations and their offspring, the bands which appered in offsprings can also occured in father or mother.5.10primers of SCoT markers for different types of persimmon germplasms can get more polymorphism on bands. The results of cluster analyses and morological traits has high uniformility. The result shows SCoT markers distinguishing persimmon germplasm is feasible, which is an effective means to researching the genetic diversity of germplasm resources of persimmon.