论文部分内容阅读
采用正交设计方法,针对影响贵州桃ISSR-PCR反应的引物浓度、模板DNA、Master Mix及退火温度等因素进行优化,建立了适于贵州桃ISSR-PCR标记的反应体系。20μL反应体系含有Master Mix 10.0μL、引物浓度0.375μmol/L、DNA模板40 ng、退火温度为52℃。利用该体系对71份贵州桃资源进行分析,扩增条带清晰,扩增产物在175~2 300 bp之间。12条ISSR引物共扩增112条带,其中多态性条带79条,多态性比率为70.54%。供试材料的Nei’s遗传多样性指数(H)为0.211 1±0.184 4,Shannon’s信息指数(I)为0.325 1±0.260 8。聚类分析显示71份贵州桃资源遗传相似系数变幅为0.669 6~0.919 6,在相似系数0.80水平处可将供试材料分为8个类群,表明贵州桃资源具有较为丰富的遗传多样性。
The orthogonal design method was used to optimize the ISSR-PCR reaction of Guizhou peach. The primers, concentration of template DNA, Master Mix and annealing temperature were optimized. 20μL reaction mixture containing Master Mix 10.0μL, primer concentration 0.375μmol / L, DNA template 40ng, annealing temperature of 52 ℃. By using this system, 71 Guizhou peach resources were analyzed, and the amplified bands were clear. The amplification products ranged from 175 to 2 300 bp. A total of 112 bands were amplified by 12 ISSR primers, of which 79 were polymorphic bands with a polymorphism ratio of 70.54%. The Nei’s genetic diversity index (H) of the tested materials was 0.211 1 ± 0.184 4, and the Shannon’s information index (I) was 0.325 1 ± 0.260 8. Cluster analysis showed that the genetic similarity coefficient of 71 Guizhou peach cultivars varied from 0.669 6 to 0.919 6, and the test materials could be divided into 8 groups at the similarity coefficient of 0.80, indicating that Peach cultivars in Guizhou are rich in genetic diversity.