利用AFLP分析西南特色猕猴桃的遗传多样性

Studies on Genetic Diversity for Specialty Kiwifruits from Southwestern Regions of China Using AFLP System

摘要:

为分析西南地区特色猕猴桃(Actinidia Lindl.)的遗传多样性,建立并优化了猕猴桃DNA 多态性分析的AFLP 体系,AFLP 标记体系为300 ng 基因组DNA 用EcoR Ⅰ/Mse Ⅰ (15 U/5 U)于37℃下酶切2 h,加接头后的混合物稀释5 倍用于预扩增,预扩增产物再稀释10 倍后进行选择性扩增.结果表明,筛选的22 对引物进行扩增反应,共得到979 条条带,其中多态性条带为649 条.采用优化的AFLP 体系,对西南地区的10 种猕猴桃50 个样本进行了遗传多样性分析,包括普通品种中华猕猴桃、美味猕猴桃,及特色品种硬毛猕猴桃、毛被猕猴桃、革叶猕猴桃、四萼猕猴桃、狗枣猕猴桃、葛枣猕猴桃、京梨猕猴桃和紫果猕猴,结果表明种内和种间的聚类关系明显；斑果组(sect. Maculatae)和净果组(sect. Leiocarpae)的种间有聚类交叉,并呈现地理区域性分布.

Abstract:

In order to understand the genetic diversity of kiwifruits in southwest of China, AFLP system for kiwifruits was optimized. The optimum AFLP system was constructed as following: 300 ng DNA was digested with EcoR I/Mse I (15 U/5 U) at 37℃ for 2 h, diluted by 5-fold after ligating with adaptors for the preamplification, and then the production of pre-amplification diluted 10-fold was used for selective amplification. The results showed that 979 electrophoretic bands were identified totally, of which 649 bands were polymorphic by using 22 pairs of primers. The genetic diversity of ten kiwifruit species, including Actinidia chinensis, A. deliciosa, A. chinensis var. hispida, A. venosa fo. pudescens, A. rubricaulis var. coriacea, A. tetramera, A. kolomikta, A. ploygama, A. callosa var. henryi and A. arguta var. purpurea, from southwestern China, were analysis by using the optimum AFLP system. The results indicated that there were obvious clustering relationship among intraspecific and interspecific, and cross cluster existed in species between sect. Maculatae and sect. Leiocarpae, related to geographic regional distribution.