茶树MFLP技术体系的建立及遗传多样性分析

Establishment of MFLP Technical System and Genetic Diversity Analysis in Tea Plant

微卫星锚定片段长度多态性(microsatellite-anchored fragment length polymorphism,MFLP)分子标记技术是结合AFLP和SSR双重原理创造的。本研究采用通用荧光引物M13-F-IRDye700(5'-CACGAC GTTGTAAACGAC-3'),利用MFLP分子标记技术建立了适合茶树的PCR反应体系,从324对引物体系中,筛选出6对最适合的引物对,对壶瓶山的44株茶树样品进行试验,分析其亲缘关系。6对MFLP选择性扩增引物共同扩增出288条带,其中多态性条带232条,多态性为80.6%,得出44株有性繁殖茶苗两两间的遗传相似系数在0.10~0.58之间,这表明同一地理位置,不同有性繁殖群体的基因组存在差异性。

Microsate ll ite-anchored fragment length polymorphism（ MFLP） molecular marker technology is created combining with AFLP and SSR dual principle. In this study, the genetic relationships for 44 genotypes of tea plant in Hu Ping mountain were analyzed by using molecular marker technology. The right PCR reaction system in tea plant was established by microsatellite-anchored fragment length polymorphism（MFLP） molecular marker technology with a universal fluorescent primer M13-F-IRDye700（5＇-CACGACGTTGTAAACGAC-3＇）adapter. Six highly polymorphic primer pairs were selected from 324 selective amplification primer combinations,and were used to analyze the genetic relationships of 44 tea samples. 288 bands were produced by 6 pairs of MFLP selective amplification primers, of which 232 bands were polymorphic, and the percentage of polymorphism was 80.6%. The genetic similarities were calculated and showed their distribution ranged from 0.10 to 0.58 among the 44 genotypes. This suggests that there are differences in the genomes of different sexual reproduction groups in the same geographical position.