Genetic Diversity And Phylogenetic Relationship Of Varieties Of Oil Tea (Camellia Oleifera Abel)

Camellia spp. is edible oil tree species native to China. The genetic researches were conducted using genomic DNA from the leaves of some varieties of C. spp. based on SSRs, AFLPs and ISSRs, respectively.SSR-PCR reaction system for C. spp. was established and optimized through orthogonal test and single-factor experiment, and eight effective primer paris were selected. The results revealed the optimal SSR-PCR reaction system in a10.0-μL volume containing DNA（5-10ng/μL）1.0μL, Mg²⁺（25mmol/L）0.7μL, dNTP （10mmol/L）0.15μL, F-and R-primer （10μmol/L）0.2μL, respectively, Taq DNA （5U/μL）0.1μL,10×PCR buffer1.0μL, HPLC H2O6.65μL. The cross-species transferability of SSR loci within genus Camellia was validated.AFLP reaction systems including restriction and ligation, pre-selective amplification and selective amplification were also established and optimized, and eight effective primer paris （E-ACT/M-AAC， E-ACG/M-CAA， E-AAG/M-CGA， E-AGT/M-AGT， E-AGG/M-AAG，E-AGG/M-AGT，E-ATC/M-AGT，E-AAT/M-CAA） were selected, which showed clearly amplified bands.The genetic diversity and phylogentic relationship of varieties of C. spp. were analyzed based on ISSRs. Higher genetic diversity was revealed for C. spp showing a percentage of polymorphic loci （PPLs） of99.25%, an expected heterozygosity （He） of0.23, Shannon’s information index （I） of0.37, while C. oleifera showed PPLs of96.24, He of0.23and I of0.37.The resultant UPGMA tree clearly separated all investigated C. spp. into three major groups corresponding to C. oleifera of Anhui, C. gigantocarpa of Guangxi and C. oleifera of Jiangxi, and generally support the traditional taxonomic classification of C. spp..In conclusion, the present study proved feasibility and usefulness of SSRs, AFLPs and ISSRs to analyse genetic diversity and phologenetic relationship of C. spp., and can also provide basic information and technical support for the conservation of genetic resource, identification of germplasm, QTL analysis and other further genetic research for C. spp. in the future.