| Rutaceae Zanthoxylum dissitum Hemsl is one of the important valuable and rare woody medicinal plant, all its root,stem and leaves,which could activate blood circulation to dissipate blood stasis, resolve toxin and disperse swelling, promote union of fracture healing,are important main raw materials for many Chinese patent drug for sprain, traumatic injury, lumbago, postpartum irregular menstruation etc. We analyse the genetic diversity among Zanthoxylum dissitum Hemsl populations by the molecular marker technology ISSR to grasp.Main research results:what the Zanthoxylum dissitum Hemsl populations’living situation is today and to decide which population is the optimum one. Those works provide a lot of theoretical basis and scientific guidance for solving the notorious Low reproduction rate problem in Zanthoxylum dissitum Hemsl planting.(1) We selecte120samples in12Zanthoxylum dissitum Hemsl populations as the experimental material. Firstly we optimize every single element by single factor experiment and decide the optimal concentration rang in every single element Secondly we preliminary screen out the preferable ISSR-PCR reaction system by orthogonal experiment and used range analysis to optimize the reaction system furtherly to get an optimal ISSR-PCR reaction system for Zanthoxylum dissitum Hemsl:20μL reaction system, template DNA30ng, Mgcl22.5mmol/L, dNTPs0.25mmol/L, primer0.7μmol/L, Taq DNA polymerase2.4U.(2) It should be noted that range analysis demonstrate only the concentration of Mg2+in the five elements has an obvious effect, which reached the significance level,towards the efficiency of the reaction system.By combining technology primer screening and annealing temperature deciding, we pick out9primers which not only have excellent repeatability but also have great polymorphism. those primers totally amplify125bands, among them124bands(the length of the bands is between200and3000bp) is varied, Percentage of polymorphic bands is99.20%,which demonstrate the level of total genetic diversity in Zanthoxylum dissitum Hemsl’s population is considerable high. Among the12populations,the the level of genetic diversity in Yangxi’(Zhangjiajie)population is the highest and in Chuanhe’(Chongqing)population is the lowest.(3) We statistically analysed the data by software PopGen32,found,at the level of species, Zanthoxylum dissitum Hemsl’s Percentage of polymorphic loci (PPB),the genetic diversity index (H) and Shannon diversity index (I) is99.20%,0.3036and0.4635respectively, which illustratethe genetic diversity of Zanthoxylum dissitum Hemsl at the level of species, which related to Zanthoxylum dissitum Hemsl itself s distribution and modes of reproduction is relatively high. The total population genetic diversity (Ht) of Zanthoxylum dissitum Hemsl is0.3036, Genetic diversity within each population is0.1673, Coefficient of genetic differentiation (Gst) is0.1363.variation within provenance account for55.10%and variation between different proven ance account for44.90%,which demonstrate the Genetic points between each population and the genetic points within a population is rough equal in12Zanthoxylum dissitum Hemsl populations. Gene flow (Nm) between each Zanthoxylum dissitum Hemsl populations is0.6138, Gene flowing between each populations is relatively lacking that shows geographical isolation occurred to a certain extent.(4) According to genetic identity and Nei’s (1978) genetic distance, we finish a NTsys2.10e clustering analysis, the result Indicates the relationship between Qingtang(Zhangjiajie)population and Yangxi(Zhangjiajie)population is the most close and the relationship between Yangxi(Zhangjiajie)population and Chuanghe(Chongqing) population is the farthestWe divide experimental materials into4types:the Ⅰ type include Qingtan(Zhangjiajie),Yangfeng(Zhangjiajie),Zoumatan(Zhangjiajie);Xiangfeng(Hubei), Hefeng(Hubei),Laifeng(Hubei) and Mushu(Guizhou) comprise the type Ⅱ; type Ⅲ consist of Changheng(Guizhou) and Xiakuag(Chongqing); typeⅣinclude Chuanghe(Chongqing) and Mawang(Chongqing). |
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