白皮松EST-SSR分子标记的开发及应用  

Development and application of EST-SSR molecular markers in Pinus bungeana

作　　者：杨雄 杨宁 袁启华 赵贵娟 李国雷[1] 贾黎明[1] 陈仲 Yang Xiong;Yang Ning;Yuan Qihua;Zhao Guijuan;Li Guolei;Jia Liming;Chen Zhong(School of Forestry,Key Laboratory of Silviculture and Conservation of Ministry of Education,Beijing Forestry University,Beijing 100083,China;Beijing Hot Spring Nursery,Beijing 100095,China;Diqing Research Institute of Forestry and Grassland Science,Diqing 674499,Yunnan,China)

机构地区：[1]北京林业大学林学院,省部共建森林培育与保护教育部重点实验室,北京100083 [2]北京市温泉苗圃,北京100095 [3]迪庆州林业和草原科学研究所,云南迪庆674499

出　　处：《北京林业大学学报》2021年第7期1-11,共11页Journal of Beijing Forestry University

基　　金：2018年北京市温泉苗圃优树种质资源圃建设项目(2018HXFWLXY008);北京市支持中央在京高校共建项目(2018GJ-03、2019GJ-03)。

摘　　要：【目的】白皮松作为我国濒危的乡土树种,具有重要的经济和园林观赏价值,为进一步开发和利用白皮松种质资源,本研究基于EST-SSR标记对北京地区3个不同种源地的白皮松群体遗传多样性开展评价。【方法】以白皮松转录组数据为依据,对其微卫星位点进行筛选并设计合成了96对引物,并对北京温泉苗圃栽培收集的3个不同种源地(北京、山东、山西)的白皮松群体,共60个植株开展遗传多样性分析,对其群体内与群体间的遗传结构进行评价。【结果】96对引物中共筛选获得了5对多态性引物,其观察杂合度、期望杂合度、多态性信息含量和等位基因位点个数变化范围分别为0.203~0.433、0.211~0.530、0.187~0.484和2~5。3个白皮松样本群体中等位基因数量、有效等位基因数量、香农信息指数、观察杂合度和固定指数变化范围为2.400~3.000、1.516~1.761、0.484~0.606、0.295~0.362和−0.075~0.081,平均值分别为2.677、1.632、0.560、0.333和−0.007。遗传分化系数和基因流变化范围分别为0.0216~0.1153和1.3996~11.3400,平均值分别为0.0902和2.5212。AMOVA分析显示遗传变异主要来自于群体内,群体间差异较小,仅占据11%。【结论】本研究共获得了5对多态性的白皮松EST-SSR引物,可用于后续白皮松群体遗传多样性分析和分子标记辅助育种工作;北京温泉苗圃栽培收集的3个不同种源地的白皮松群体分析结果表明,当地现有收集的白皮松种源群体遗传相似度较高,在未来种质资源保存和苗木繁育工作中应考虑增加其他种源地的白皮松群体。[Objective]As an endangered native tree species in China,Pinus bungeana has important economic and garden ornamental values.In order to further develop and utilize the species germplasm resources,the study evaluated the genetic diversity of three populations from different provenances in P.bungeana.[Method]Simple sequence repeats were selected from the transcriptome data in P.bungeana,and then 96 pairs of primers were developed,and the genetic diversity of 60 individuals in Beijing Hot Spring Nursery collected from three different provenances(Beijing,Shandong,and Shanxi)was analyzed,and genetic diversity parameters between intra-and inter-population were analyzed.[Result]The variation ranges of the observed heterozygosity,expected heterozygosity,polymorphism information content and the number of allele loci for 5 pairs of polymorphic primers were 0.203−0.433,0.211−0.530,0.187−0.484 and 2−5,respectively.The number of alleles,effective alleles,Shannon diversity index,observed heterozygosity and fixed index of the three populations ranged in 2.400−3.000,1.516−1.761,0.484−0.606,0.295−0.362 and−0.075−0.081,the average values were 2.677,1.632,0.560,0.333 and−0.007,respectively.The ranges of genetic differentiation coefficient and gene flows were 0.0216−0.1153 and 1.3996−11.3400,and the average values were 0.0902 and 2.5212,respectively.AMOVA analysis showed that genetic variation mainly comed from within the population,with small differences between populations,accounting for only 11%.[Conclusion]This study obtained 5 pairs of polymorphic EST-SSR primers in P.bungeana,which can be used for subsequent analysis of the population genetic diversity and molecular marker-assisted breeding.Genetic diversity analysis about the three populations of P.bungeana shows that the existing populations in Beijing Hot Spring Nursery have high genetic similarity,and the populations from other provenances should be considered for the future germplasm resource preservation and seedling breeding work.

关 键 词：白皮松 EST-SSR 引物开发 遗传多样性评价 群体遗传结构 

分 类 号：S791.243[农业科学—林木遗传育种]