RAPD Analysis And In Vitro Conservation Of Germplasm Resources Of Oolong Tea (Camellia Sinensis)

There are lots of cultivars in Fujian Province. The genetic diversity and relationship of thirty varieties of oolong tea plants from Fujian Province was studied based on RAPD molecular markers. The plants Jin Fenghuang were applied as the conservation materials to investigate the restricting growth conservation method .The main results were as follows:1. It is difficult to extract DNA from tea plant because of phenolic compound and quit a lot of polysaccharose .Based on the previous research, the method about extraction and purification of tea genetic DNA are simple and accurate. The researcher used improved CTAB-mini method to extract DNA, which meet RAPD-PCR requirements in quantity and purity.2. 11 primers were selected from 37 random primers and were used to analyze by using RAPD among 30 accessions, which generated 125 bands of which polymorphic bands number was 108, the percentage of polymorphism equaled 86.40% .The similarity coefficient was calculated using the Nei coefficient method by NTsys 2.10e software. The results showed that the coefficient among 30 accessions ranged from 0.41 to 0.83, indicating the gene differentiation was very remarkable among tested cultivars.3. According to the clustering analysis of RAPD results. Based on the similarity coefficient, a dendrogram was constructed to indicate their genetic diversity. From the result, genetic similarity and relationship were revealed. When similarity coefficient was 0.40,30 accessions of germplasm resource of tea plant could be separated into 3 groups, Xiaoqi consist one group. It proved to use RAPD for identification and classification of accessions within Resource of TEA Plant .It laid theoretical basis for introduce of breeding , shorten the breeding and improved the efficiency of breeding.4. In the study of germplasm preservation of oolong tea with different concentrations of mannitol, sucrose and PP₃₃₃ influenced preservation time .Results showed that the best medium was 1/2MS supplemented with mannitol 15 g/L. medium supplemented with sucrose 30 g/L or PP₃₃₃ 20 mg/L or ABA 0.5 mg/L were good for the germplsam presevation of tea plant.