文献类型：期刊文献

中文题名：基于cp DNA的当归野生与栽培种质鉴定与遗传变异分析

英文题名：Identification and Genetic Variation Analysis of Wild and Cultivated Germplasm of Angelica sinensis Based on cp DNA

作者：张明惠[1];朱田田[1,2,3];晋玲[1,2,3];王富胜[4];康舒淇[1];徐丽[1];王圆圆[1]

第一作者：张明惠

机构：[1]甘肃中医药大学,兰州730000;[2]西北中藏药省部共建协同创新中心,兰州730000;[3]甘肃省珍稀中药资源评价与保护利用工程研究中心,兰州730000;[4]定西市农业科学研究院,甘肃定西743000

第一机构：甘肃中医药大学

年份：2022

卷号：28

期号：15

起止页码：129

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：甘肃省科技厅创新基地和人才计划目(20JR5RA182);道地药材生态种植及质量保障项目(国中医药科技[2020]153号);甘肃省教育厅“双一流”科研重点项目(GSSYLXM-05);甘肃中医药大学科学研究与创新基金项目(2021KCZD-4);中国工程院战略研究与咨询项目(GS2021ZDA06-2)。

语种：中文

中文关键词：当归;叶绿体基因(cpDNA);野生;栽培;遗传变异

外文关键词：Angelica sinensis;chloroplast DNA(cp DNA);wild;cultivated;genetic variation

摘要：目的:基于叶绿体基因(cp DNA)对甘肃省11个当归栽培品种(系)及7个野生当归居群进行了遗传变异分析,为当归的种质鉴定和新品种的选育提供参考。方法:利用3对cp DNA引物对当归样品进行聚合酶链式反应(PCR)扩增并测序,利用MegaX软件对序列特征进行统计,并计算当归居群间平均遗传距离,利用NTSYS 2.10e软件构建遗传距离非加权组平均法(UPGMA)聚类图。利用DanSP v6软件计算当归序列多态性及Tajima中性检验;利用PERMUT软件计算当归居群结构;利用Arlequin v3.5软件进行分子变异分析;最后利用PopART 1.7软件构建TCS单倍型网络图。结果:3对cp DNA引物扩增、测序、比对、合并后的序列长度为1759 bp,野生当归检测到1个变异位点,栽培当归检测到480个变异位点,其中单一突变位点97个,简约信息位点383个,插入-缺失位点152个。在野生组和栽培组当归cp DNA的matK、psbA-trnH和rbcL 3个区域中,多态性最高的区域均为matK。全部当归联合序列的Tajima中性检验均为不显著负值,但在栽培当归中psbA-trnH和rbcL基因中呈显著负值,说明当归整体上遵循中性进化,而psbA-trnH和rbcL基因经历过选择。野生居群间的遗传分化程度(Fst=0)低于栽培当归居群间的分化程度(Fst=0.11419,P<0.05),且二者之间遗传分化程度较高(Fst=0.94255,P<0.01)。栽培当归居群中变异主要来源于居群内部(89%)。遗传距离UPGMA聚类树显示野生当归和栽培当归各自聚为一支,亲缘关系较远,栽培当归中居群3距离其他栽培当归居群较远。TCS单倍型网络图由15个单倍型和4个未知单倍型构成,分为3部分,各部分间变异次数较多,共享单倍型仅分布于野生或栽培组之内,组间不存在共享单倍型。结论:当归在物种水平上遗传多样性偏低,野生当归居群多样性低于栽培当归,野生与栽培当归组间的遗传分化程度高,而野生当归居群内和栽培当归居群内的分化程度均较低,栽培当归中遗传变异主要存在于居群内。
Objective To conduct genetic variation analysis of 11 cultivars and 7 wild populations of Angelica sinensis in Gansu province based on the chloroplast gene(cp DNA),and provide references for germplasm identification and breeding of new cultivars of A.sinensis.Method Three pairs of cp DNA primers were used for polymerase chain reaction(PCR)amplification and sequencing of A.sinensis samples.MegaX was used to perform statistics on sequence characteristics and calculate mean genetic distances among A.sinensis populations.Unweighted pair-group method with arithmetic means(UPGMA)clustering tree based on genetic distance was constructed by NTSYS 2.10e.DanSP v6 was used to calculate sequence polymorphism and Tajima's D of A.sinensis.PERMUT was used to calculate the population structure of A.sinensis.Arlequin v3.5 was used to perform molecular variation analysis,and PopART1.7 was used to construct TCS haplotype network.Result Three pairs of cp DNA primers were amplified,sequenced,compared,and combined to give a sequence length of 1759 bp.One variable site was detected in the wild A.sinensis and 480 variable sites were detected in the cultivated A.sinensis,including 97 singleton variable sites,383 parsimony informative sites,and 152 insertion-deletion sites.In the three regions of matK,psbA-trnH,and rbcL of cp DNA in the wild and cultivated A.sinensis,matK was the region with the highest polymorphism.Tajima’s D of all the combined sequences of A.sinensis were not significantly negative,but psbA-trnH and rbcL genes of the cultivated A.sinensis were significantly negative,indicating that the A.sinensis followed neutral evolution on a whole,while psbA-trnH and rbcL genes had undergone selection.The degree of genetic differentiation(Fst=0)among wild populations was lower than that among cultivated populations(Fst=0.11419,P<0.05).The degree of genetic differentiation between wild and cultivated A.sinensis was relatively high(Fst=0.94255,P<0.01).Genetic variation in the cultivated A.sinensis was mainly found within the populations(89%).UPGMA cluster tree based on genetic distance showed that the wild A.sinensis and the cultivated A.sinensis were clustered into one branch,respectively,with a distant genetic relationship,and the population 3 in the cultivated A.sinensis was far from other cultivated populations.The TCS haplotype network consisted of 15 haplotypes and 4 unknown haplotypes,which was divided into 3 parts,with a large number of variations among each part.Shared haplotypes were only found in the wild or cultivated groups,and there were no shared haplotypes between groups.Conclusion The genetic diversity of A.sinensis was low at species level,and the population diversity of the wild was lower than that of the cultivated.The degree of genetic differentiation between the wild and the cultivated A.sinensis was high,but that in the wild and the cultivated populations were low.Genetic variation in the cultivated A.sinensis was mainly found within populations.