| The genetic diversity on 32 cross-vigor strains of Camellia oleifera (such as Min 1) and the 10 cultivars from Camellia brevistyla were studied by random amplified polymorphic DNA (RAPD), and the relationship between the genetic character and the marks of RAPD were analyzed .The thesis was composed of 4 parts. Part I: Extraction of DNA from the fresh leaves of oil camellia; Part II: Study on RAPD reaction condition for oil camellia. Part III: Genetic diversity among 42 samples analysis. Part IV: Analysis on the relationship between the genetic character and RAPD marks. The results showed that: (1) The isolated DNA by the improved method with CTAB was 48kb, and OD260/OD280 of DNA was 1.8~2.0. The DNA average yield was 328 ng/(mg · fresh weight).Thc isolated DNA was suitable for digesting with restriction enzyme. (2) Effects of the content of DNA templates, Mg2+, dNTPs, primers and Taq DNA polymcrasc on experimental results were tested. The results showed that each 25 μl amplification reaction solution was consisted of 20ng template DNA, 2.5mmol/L MgCl2, 0.2mmol/L dNTPs, 0.3 μmol/L primer and IU Taq DNA polymerase. The amplification procedure conditions were pre-denature at 94℃180s followed by denature 94℃ 60s, annealing 37℃ for 90s, extension 72℃ for 120s, cycling 40 times, last extension 72℃ for 5min, and then the optimum procedure for oil camellia DNA was obtained.(3)A total of 159 repeatable loci (including 94 polymorphic loci) were amplified using 19 random primers. The percentage of polymorphic bands (PPB) was 59.12%.As analyzed by POPGENE version 1.32, the data from the 42 samples had an average Shannon's information index of 0.2516, and Nei's gene diversity index of 0.1653. The results suggested that the genetic diversity was relatively low among these samples. And it also indicated that the genetic distance between Camellia oleifera and Camellia brevistyla is 0.2111. (4) Relationships among 32 cross-vigor strains of Camellia oleifera based on their Nei's unbiased distances were clustered using unweighted group method arithmetic average (UPGMA) analysis in a dendrogram which divided the 32 strains into 5 groups (A, B, C, D, E). The dendrogram indicated that the content of tea-oil in the seed in the strains of group A is higher than other groups, and the strains which had the lowest content of tea-oil in the seed were included into group C. The results suggested that the genetic character of the content of tea-oil in the seed may relate to some RAPD marker. |
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