69份苦玄参种质SSR遗传多样性及品质性状相关标记分析  

SSR-based Analysis of Genetic Diversity and Quality Trait-related Markers of 69 Picria felterrae Germplasm Samples

作　　者：闫国跃 杨帆 白燕远 李耀燕 吴梦丽 高慧 张淼 阿里穆斯[1,4] 谢阳姣 YAN Guo-yue;YANG Fan;BAI Yan-yuan;LI Yao-yan;WU Meng-li;GAO Hui;ZHANG Miao;BORJIGIDAI Almaz;XIE Yang-jiao(Key Laboratory of Ethnomedicine,Ministry of Education,School of Pharmacy,Minzu University of China,Beijing 100081,China;College of Yao Medicine,Guangxi University of Chinese Medicine,Nanning 530001,China;Guangxi Talent Highland for Zhuang and Yao Medicine and Combination of Medical Care and Elderly Care,Nanning 530001,China;Guangxi Center for Zhuang and Yao Medicine Engineering and Technology Research,Nanning 530200,China;Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine Collaborative Innovation Center of Zhuang and Yao Ethnic Medicine,Nanning 530200,China)

机构地区：[1]中央民族大学民族医药教育部重点实验室,药学院,北京100081 [2]广西中医药大学瑶医药学院,南宁530001 [3]广西壮瑶医药与医养结合人才小高地,南宁530001 [4]广西壮瑶药工程技术研究中心,南宁530200 [5]广西壮瑶药重点实验室(壮瑶药协同创新中心),南宁530200

出　　处：《中国实验方剂学杂志》2020年第4期174-184,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(31460074,81573535);国家重点研发计划项目(2017YFC1704000);民族医药教育部重点实验室自主课题(KLEM-ZZ201805);广西壮瑶药重点实验室项目(桂科基字【2014】32号);壮瑶药协同创新中心项目(桂教科研【2013】20号)。

摘　　要：目的:采用简单重复序列(SSR)分子标记技术对广西苦玄参主产区69份苦玄参种质样本进行遗传多样性及亲缘关系分析,并筛选与苦玄参苷含量相关联的优良种质基因。为苦玄参种质资源评价、遗传进化分析及分子标记辅助育种等提供依据。方法:基于转录组测序技术,开发20对引物进行批量扩增。利用各标记位点的遗传多态信息,分析69份苦玄参种质的遗传多样性及亲缘关系,并通过一元线性和多元逐步回归分析,筛选与苦玄参苷含量相关联的分子标记。结果:20对SSR引物共扩增出76个等位基因,平均每个位点观测等位基因3.8个,高于有效等位基因数(1.9692个),稀有等位基因率为38.2%,等位基因分布不均匀。等位基因多态率范围为0~59%,平均38.24%,各位点多态率差异较大。各位点多态信息含量(PIC)变化范围为0~0.6211,平均0.3780;Shannon多态性信息指数变化范围为0~1.2401,平均0.7590;Nei’s基因多样性指数(Nei)变化范围为0~0.6823,平均0.4409;以上3个指标最高的为P21,最低为P7,各位点遗传多样性存在较大差异。各位点平均观测杂合度为0.3824,低于平均期望杂合度(0.4425),表现为杂合子缺失;平均遗传分化系数Fst为0.3659;基因流Nm平均值为0.4332,种质遗传分化较大,基因流较小。一元线性回归分析和多元逐步回归分析结果表明,与苦玄参苷IA和IB相关的位点各有5个,其中仅有1个位点与2个成分的含量均相关。结论:20个SSR标记位点遗传多样性存在较大的差异,供试69份种质遗传分化大,基因流较小;从供试20个SSR标记中筛选出9个与苦玄参苷含量相关联的标记位点,试验结果可为苦玄参遗传进化分析及良种选育和繁育等提供依据。Objective:Sixty-nine germplasm samples of Picria felterrae collected from the main producing areas in Guangxi were subject to genetic diversity and genetic relationship analyses using the simple seguence repeat(SSR)molecular marker technology and good germplasm genes associated with the content of picfeltarraenins were screened so as to provide references for germplasm resource evaluation,phylogenetic analysis,and molecular mark assisted breeding of that species.Method:20 pairs of randomly selected primers were amplified based on the transcriptome sequencing technology.The genetic diversity of and genetic relationship between the 69 samples were analyzed using the genetic polymorphic information for each marker locus,and one-variable linear regression and multiple stepwise regression analyses were performed to screen molecular markers associated with the content of picfeltarraenins.Result:The amplification using the 20 pairs of SSR primers produced 76 alleles,3.8 alleles for each locus on average,higher than effective alleles(1.9692),and the rare allele rate was 38.2%,suggesting that the alleles distributed unequally.The polymorphism rates of alleles varied between 0-59%,with an average of38.24%,showing a great difference among loci.The polymorphic information content(PIC)varied between 0-0.6211,with an average of 0.3780.Shannon polymorphic information index varied between 0-1.2401,with an average of 0.759.Nei’s gene diversity index(Nei)varied between 0-0.6823,with an average of 0.4409.P21 had the highest level accompanied with the lowest P7 for the above three indexes,and significant genetic diversity differences were identified among the loci.For all loci,the mean observed heterozygosity was 0.3824,lower than the average expected heterozygosity of 0.4425,suggesting the loss of heterozygosity,the average genetic differentiation coefficient(Fst)was 0.3659 and the average gene flow Nm was 0.4332,suggesting a high genetic divergence and a low gene flow.The results of one-variable linear regression and multiple st

关 键 词：苦玄参 简单重复序列(SSR)分子标记 遗传多样性 品质性状 相关分析 

分 类 号：R284.2[医药卫生—中药学] R289[医药卫生—中医学]