Genetic Diversity Of Analysis Taxus L.Resources And Establishment Of Callus Induction In Taxus Var. Mairei

Taxus L. is the first Class preserved plant in China andit contains taxol, an important anticancer, which made it very difficult to preserve Taxus L. resources. In order to utilize and preserve Taxus L. resources effectively, the genetic diversity of Taxus var. mairei from Liangye National Natural Protection District was analysed and was compared with Taxus wallichiana and Taxus Chinensis var. mairei. The callus induction system of Taxus var. mairei was also established in the expriment, the main results were as following:1. Buds was the best explant for callus induction. 2. Medium B5 adding 2. Omg/L 2, 4-D , 1. Omg/L NAA . 1. Omg/L IAA, 2. Og/L LH and 0. 5g/L AC was the most optimized medium for callus induction. 3. Medium B5' containing 4. Omg/L NAA. 0. 5mg/L 2, 4-D. 0. 2mg/L GA3. 0. 5g/L AC and 2. Og/L LH was the most favorable for callus growth. The brownness of callus was avoided or mitigated to some degree when AC. LH and GA3 were added to the culture medium together. 4. The parameters of RAPD analysis were optimized for Taxus L, the total volume of 25M-1 containing 200Wnol/L dNTP, 0. 2Pmol/L random primers. 30- 100ng template DNA and 1. 5 unit Taq DNA enzyme. The program for the PCR amplification was that first denatured the template DNA at 94癈for 3.0 min and then denatured the DNA at 94癈 for 1.0 min, annealed at 36癈 for 2.0 min, extended at 72癈 for 2. 0 min, 45 cycles, then extended at 72癈 for10. 0 min and kept the reaction at 4癈 until it was taken out. 5. The total of 398 DNA bands were amplified in the eleven Taxus L. resources by the above PCR procedure using 9 random primers, and about 27% of the bands were polymorphic, indicating that the genetic background was narrow in Taxus L. involved in this experiment. 6. An cluster analysis was made by using the RAPD data of Taxus L . involved in the experiment, and the results showed that the Taxus L. resources were divided into 2 subclasses when the value of D was 0. 52:the first subclass was the cultivars of Taxus Media var. and Taxus wallichiana , the second was Taxus Chinensis var.mairei; Taxus L. resources were divided into 3 subclasses when the value of D was 0.67,apart from the above first subclass , the second subclass was further distinguished between cultivars of Taxus Chinensis var.mairei and wild varieties of Taxus Chinensis var.mairei, indicating that genetic differences among the subclasses of Taxus Chinensis var.mairei; when the value of D was 0. 80, some genetic differences were showed among Taxus Chinensis var.mairei.