应用FISH-AFLP技术分析平欧杂种榛主栽品种的遗传关系

Analysis of the Genetic Relationship of the Main Cultivars of Ping'ou Hybrid Hazelnut(C. heterophylla×C. avellana) by FISH-AFLP Markers

【目的】建立平欧杂种榛FISH-AFLP技术体系,并应用该技术分析平欧杂种榛主栽品种的遗传关系。【方法】以达维等10个主栽品种为试材,经DNA提取、PstⅠ/MseⅠ双酶切,进行FISH-AFLP反应,数据转换为"0—1"矩阵后,使用NTSYS pc 2.11F和Popgene 1.32软件进行数据分析并作图。【结果】从64对引物中,筛选出15对多态性强的PstⅠ/MseⅠ引物,共获得1 739条谱带,引物平均多态带比率97.94%;平欧杂种榛主栽品种的相似性系数为0.7556—0.8543,当阈值为0.8398时,可分成4个AFLP群,其中,玉坠(84-310)、辽榛4号(85-41)和平欧69号(84-69)单独为1群,其他品种为1群;平欧杂种榛主栽品种的有效等位基因数、基因多样度、Shannon信息指数分别为1.3921、0.2482和0.3957,具有较高的遗传多样性;研究获得的特征条带可用于平欧杂种榛主栽品种的快速鉴别。【结论】由于平欧杂种榛的实生选优群体是多亲本组合的混合后代,因此,该群体是一个具有高度遗传多样性的群体,主栽品种间存在复杂的亲缘关系;研究构建的FISH-AFLP技术流程、筛选得到的引物以及分析得到的各项遗传参数,可为其他榛属植物的相关研究提供参考。

[Objective] The objective of this study is to establish FISH-AFLP analysis system for Ping＇ou hybrid hazelnut and analyze the genetic relationship of the main cultivars. [Method] Ten main cultivars, such as Dawei, were selected as materials. High-quality genomic DNA was extracted with improved CTAB method, and the purified DNA samples were digested with Pst I /Mse I. After ligation reaction, the samples were used to perform FISH-AFLP detection. The information of all bands was converted into ＂0-1＂ matrix. NTSYS pc 2.11F and Popgene 1.32 software were used to conduct the data analysis and plotting. [Result] The results showed that： 15 Pst I/Mae I primer pairs were selected from 64 combinations with high polymorphism, strong signals and clear bands. A total of 1 739 bands were obtained. The average percentage of polymorphic bands was 97.94%. The similarity coefficient of the main cultivars varied from 0.7556 to 0.8543. All the samples were divided into 4 groups when the threshold value was 0.8398. Each in cutivars of Yuzhui （84-310）, Liaozhen No. 4 （85-41） and Ping＇ou 69 （84-69） formed an independent group alone and the others constituted the fourth group. The effective number of alleles, Nei＇s gene diversity and Shannon＇s information index of the main cultivars were 1.3921, 0.2482 and 0.3957, respectively, indicating high genetic diversity among Ping＇ou hybrid hazelnut. The unique bands obtained in the experiments could be used in the rapid identification of the main cultivars of Ping＇ou hybrid hazelnut. [Conclusion] Due to the complex selecting group, Ping＇ou hybrid hazelnut is a high polymorphic population and the genetic relationships of the main cultivars are complex. The FISH-AFLP analysis system, Pst I/Mse I primer pairs and all the genetic parameters obtained in this research can offer reference for the relative researches in Corylus.