基于SRAP标记的紫溪山华山松种子园无性系遗传多样性分析

SRAP marker analysis of genetic diversity in Pinus armandii clones from seed orchard in Zixi Mountain

为明确云南楚雄市紫溪山华山松种子园内不同种源无性系间的遗传背景,该研究收集了园内6个种源的60个华山松无性系单株针叶,采用改良CTAB法提取其总DNA,并利用SRAP分子标记对其进行遗传多样性分析。结果表明:(1)从100对引物组合中共筛选出15对具有多态性的SRAP引物,经SRAP-PCR扩增后,共获得出194个位点,多态位点百分率(PPB)为85.05%,Nei’s基因多样性指数(H)为0.2337,Shannon’s信息指数(I)为0.3419,种源间的遗传分化系数(GST)为0.3555。(2)华山松6个种源遗传多样性较高,且遗传变异主要存在于华山松种源内,种源地会泽(HZ)与巍山(WS)种源的遗传距离最近(D=0.0501),会泽(HZ)与宜良(YL)种源的遗传距离最远(D=0.3618)。(3)聚类分析显示将6个华山松种源一共聚为3类:会泽(HZ)和巍山(WS)种源聚为一类;楚雄(CX)、南华(NH)和宜良(YL)种源聚为一类;腾冲(TC)种源单独为一类。综合上述结果显示,紫溪山华山松种子园内无性系间的遗传分化处于较高水平,为华山松杂交育种时亲本的选配及种质资源的评价提供了分子水平的理论依据。

In order to clear genetic background of different provenances Pinus armandii colnes from seed orchard in Zixi Mountain of Chuxiong City,Yunnan Province,the needles of 60 clones were taken from six provenances which was used to extract total DNA by modified CTAB method,and the genetic diversity was analyzed by SRAP molecular markers.The results were as follows:(1) Fifteen pairs of polymorphic SRAP primers were screened from 100 pairs of primer combinations.After SRAP-PCR amplification,a total of 194 loci were obtained,and the percentage of polymorphic loci PPB =85.05%,Nei’s gene diversity index H = 0.233 7,Shannon’s information index I = 0.341 9,genetic differentiation coefficient between population GST= 0.355 5.(2) The genetic diversity of 6 provenances of P.armandii was high,and the genetic variation was mainly existed within provenances of P.armandii.The genetic distance between the Huize(HZ)provenance and the Weishan(WS) provenance was the closest(D = 0.050 1),that the Huize(HZ) provenance and Yiliang(YL) provenance was the farthest(D = 0.361 8).(3) The cluster analysis showed that six P.armandii provenances there were classified into three categories: Huize(HZ) and Weishan(WS) provenances were in one category;Chuxiong(CX),Nanhua(NH) and Yiliang(YL) provenances were in the second category;Tengchong(TC) provenance was a separate category.The results indicate that genetic differentiation in P.armandii clones from seed orchard in Zixi Mountain of Chuxiong City has a high level,and it can provide theoretical information and practical guidance for parent selections during the cross breeding and evaluation of germplasm resources of P.armandii as well.