安徽砀山酥梨自然保护区梨种质资源AFLP分析

Study on the pear germplasm resources in Dangshan area,Anhui province by AFLP technology

为探讨安徽砀山酥梨自然保护区梨种质资源遗传多样性和鉴定砀山酥梨营养系变异,以该保护区内27个梨资源代表类型为试材,利用AFLP分子标记技术开展研究。运用从36对选择性扩增引物中筛选的3对引物组合进行扩增,每对引物可扩增出50条以上的清晰条带,扩增片段长度为30～600bp,9个引物组合共扩增出清晰可靠的DNA谱带540条。其中498条为多态性条带,平均多态性比率为92.2%。聚类分析表明,当遗传距离阈值为39.2～40.1时,全部供试生物型可分为6类:第1类为砀山酥梨及其变异类型和原产砀山的地方品种、杂交种;第2类包括雪花梨、鸭梨、紫酥、慈梨、早美酥、丰水、新高,白梨和砂梨聚在了一类;第3类包括库尔勒香梨、红香酥;南果梨、红巴梨、杜梨各为一类。与砀山酥梨相比,砀山酥梨的8个营养系变异类型均扩增出了特异带,特异条带出现的数量为5～15条不等。说明安徽砀山酥梨自然保护区梨种质资源遗传多样性丰富;砀山酥梨8个营养系变异类型均在DNA水平发生了不同程度的突变。

Total of 27 biotypes of pear, cultivated in the Protection District of Dangshansu Pear, Dangshan County, Anhui Province were analyzed by the established technical system of amplified fragment length polymorphism （AFLP）for studying the genetic diversity and mutant＇ s identification in genus Pyrus. Three pairs of selective primer screened out from 36 pairs of selective ones were used in the amplification and more than 50 bands were amplified by each pair of selective primer. The size of amplified fragment was 30 to 600 bp, 540 bands amplified by 9 primer combinations was reliable in this AFLP technique system, and the 498 bands of which were polymorphic accounting for 92.2% of total bands in the genus. The result of cluster analysis showed that all tested biotypes could be divided into 6 groups at threshold of 39.2 to 40.1 in coefficient of squared euclidean distance. The first group was composed of Dangshansu pear and its 8 mutants, intraspecies hybrid and some local cultivars. The second group included Xuehuali, Yali, Cili, Zisu, Zaomeisu, Hosui and Niitaka, where P. bretschneideri Rehd and P. pyrifolia Naka cluster into the same group. The third group was made up of Kuerlexiangli and Hongxiangsu. Nanguoli, Red Bartlett and P. betulaefolia became a separate group respectively. Compared with the stock plant Dangshansu pear the special fragment has been amplified in all mutants and the number of special band was unequally from 5 to 15. It was proved that the genetic diversity was rich in the Protection District of Dangshansu Pear and there was genetic variation happened in the genomic DNA of the mutants.