基于EST-SSR标记的珠子参野生种质资源遗传多样性分析
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摘要
【目的】利用EST-SSR标记分析云南省珠子参野生资源的遗传多样性。【方法】利用17对人参属EST-SSR引物对云南省珠子参主要分布区的19个珠子参野生居群进行遗传多样性分析。【结果】在物种水平,17对人参属EST-SSR引物在19个珠子参居群中共检测到346个位点,其中多态位点296个,平均为17. 4个,珠子参的等位基因数(A)为1. 8555,Nei’s基因多样性(H)为0. 2023、Shannon指数(I)为0. 3230;在居群水平,19个居群的等位基因数(A)为1. 0954～1. 2977,Nei’s基因多样性(H)为0. 1043～0. 1387,Shannon指数(I)为0. 0309～0. 1077,多态位点数(PPB)为8. 67%～29. 77%。居群间的遗传分化系数(Gst)为0. 6357,居群每代迁移数(Nm)为0. 2866。遗传相似度和聚类分析的结果表明,19份珠子参居群被划分为2个大类群,其中第二类群16个居群被分为4个小类群。【结论】珠子参总体上具有丰富的遗传多样性,但各个居群的遗传多样性不高,居群间缺乏基因交流;居群间遗传分化水平高,遗传差异主要存在于居群间;聚类分析显示,不同表型的珠子参虽然形态差异较大,但遗传差异不大。
【Objective】Genetic diversity of wild Resources P. japonicus in Yunnan province by EST-SSR markers were analyzed.【Method】The genetic diversity and genetic structure of 19 P. japonicas populations in the main distribution area of Yunnan province were analyzed by using 17 pairs of EST-SSR molecular markers. 【Result】In species level,a total of 346 mutation sites were detected in 19 populations of by17 EST-SSRs primer pairs from Panax,among which 296 were polymorphic loci. The allele( A) number was 1. 8555 in P. japonicus,the Nei's( H) was 0. 2023,and Shannon's information index( I) was 0. 3230. In population level,the allele was 1. 0954-1. 2977 in 19 populations,Nei's gene diversity( H) was from 0. 1043 to 0. 1387 and the range of Shannon's information index( I) was from 0. 0309 to0. 1077. Total percentage of polymorphic loci( PPB) was from 8. 67 % to 29. 77 % in these populations. Coefficient of gene differentiation( Gst) was 0. 6357 among populations of P. japonicus. And the populations transport number( Nm) of one generation was 0. 2866. Genetic similarity and cluster analysis results showed that 19 populations of P. japonicus were divided into two large groups,among which 16 populations could be classified into 4 groups.【Conclusionl】P. japonicus had more genetic diversity,but had low genetic diversity in each populations and was lack of genetic communication among populations. The level of genetic differentiation among populations was higher,and the genetic difference was mainly existed among populations. Although the phenotypic diversity in different populations was great,but the genetic diversity was small.
【Objective】Genetic diversity of wild Resources P. japonicus in Yunnan province by EST-SSR markers were analyzed.【Method】The genetic diversity and genetic structure of 19 P. japonicas populations in the main distribution area of Yunnan province were analyzed by using 17 pairs of EST-SSR molecular markers. 【Result】In species level,a total of 346 mutation sites were detected in 19 populations of by17 EST-SSRs primer pairs from Panax,among which 296 were polymorphic loci. The allele( A) number was 1. 8555 in P. japonicus,the Nei's( H) was 0. 2023,and Shannon's information index( I) was 0. 3230. In population level,the allele was 1. 0954-1. 2977 in 19 populations,Nei's gene diversity( H) was from 0. 1043 to 0. 1387 and the range of Shannon's information index( I) was from 0. 0309 to0. 1077. Total percentage of polymorphic loci( PPB) was from 8. 67 % to 29. 77 % in these populations. Coefficient of gene differentiation( Gst) was 0. 6357 among populations of P. japonicus. And the populations transport number( Nm) of one generation was 0. 2866. Genetic similarity and cluster analysis results showed that 19 populations of P. japonicus were divided into two large groups,among which 16 populations could be classified into 4 groups.【Conclusionl】P. japonicus had more genetic diversity,but had low genetic diversity in each populations and was lack of genetic communication among populations. The level of genetic differentiation among populations was higher,and the genetic difference was mainly existed among populations. Although the phenotypic diversity in different populations was great,but the genetic diversity was small.
引文
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[14]王薇,宋小妹,康亚国,等.大孔吸附树脂富集纯化珠子参总皂苷的工艺研究[J].辽宁中医杂志,2010(4):713-715.
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[17]赵毅,赵仁,宋亮,等.珠子参药材品种概述及资源现状调查[J].中国现代中药,2011(1):11-17.
[18]刘万里,刘婷,何忠军,等.珠子参规范化栽培技术[J].陕西农业科学,2014(8):127-128.
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[20]杨成君,王军,穆立蔷,等.人参EST-SSR标记的建立[J].农业生物技术学报,2008(1):114-120.
[21]Charters Y M,Wilkinson M J. The use of self-pollinated progeniesas‘in-groups’for the genetic characterization of cocoa germplasm[J]. Theoretical and Applied Genetics,2000,100:160-166.
[22]崔光芬.云南铁皮石斛和齿瓣石斛DALP和ITS遗传变异研究[D].昆明:云南大学,2007.
[2]杨文鹏,关琦,杨留启,等.贵州70份玉米自交系的SSR标记遗传多样性及其杂种优势群分析[J].植物遗传资源学报,2011,12(2):241-248.
[3]张云,龙云星,奎丽梅,等.我国水稻两用核不育系SSR遗传多样性分析[J].分子植物育种,2017,15(7):2836-2846.
[4]杨旺兴,卓伟,马彬林,等. SSR标记遗传距离与水稻籼粳衍生系杂种优势的相关性分析[J].西南农业学报,2017,30(6):1251-1256.
[5]李为民,王宇超,黎斌,等.陕西省野生大豆种质资源的SSR遗传多样性研究[J].中国农学通报,2015,31(24):99-105.
[6]王彪.中国栽培大豆遗传多样性SSR研究[D].杨凌:西北农林科技大学,2002.
[7]袁力行,傅骏骅,Warburton M,等.利用RFLP、SSR、AFLP和RAPD标记分析玉米自交系遗传多样性的比较研究[J].遗传学报,2000(8):725-733+756.
[8]温莹,逯晓萍,任锐,等.高丹草EST-SSR标记的开发及其遗传多样性[J].遗传,2013,35(2):225-232.
[9]杨佳.蜡梅EST-SSR分子标记的开发及其遗传多样性,遗传结构的研究[D].西安:西北大学,2012.
[10]张金渝,杨维泽,崔秀明,等.三七栽培居群遗传多样性的EST-SSR分析[J].植物遗传资源学报,2011,12(2):249-254.
[11]宋振巧,王建华,陈为序,等. EST-SSR分析不同产地丹参的遗传多样性[J].核农学报,2014,28(2):193-199.
[12]陈庚,姜薇薇,贾兵,等.珠子参根状茎中不同部分的皂苷组成分析[J].中药材,2018(7):1647-1651.
[13]李凤英,王英姿,王晴,等.正交试验法优化珠子参总皂苷提取工艺[J].北京中医药大学学报,2016(2):151-154.
[14]王薇,宋小妹,康亚国,等.大孔吸附树脂富集纯化珠子参总皂苷的工艺研究[J].辽宁中医杂志,2010(4):713-715.
[15]崔九成,张旋,杨新杰,等.不同产地珠子参中微量元素含量分析[J].光谱实验室,2011(6):2866-2870.
[16]李利霞,赵厚涛,朱虹,等.珍稀濒危植物珠子参研究进展[J].陕西农业科学,2015,61(2):59-61+71.
[17]赵毅,赵仁,宋亮,等.珠子参药材品种概述及资源现状调查[J].中国现代中药,2011(1):11-17.
[18]刘万里,刘婷,何忠军,等.珠子参规范化栽培技术[J].陕西农业科学,2014(8):127-128.
[19]邹喻苹,葛颂,王晓东.系统与进化植物学中的分子标记[M].北京:科学出版社,1998:16-25
[20]杨成君,王军,穆立蔷,等.人参EST-SSR标记的建立[J].农业生物技术学报,2008(1):114-120.
[21]Charters Y M,Wilkinson M J. The use of self-pollinated progeniesas‘in-groups’for the genetic characterization of cocoa germplasm[J]. Theoretical and Applied Genetics,2000,100:160-166.
[22]崔光芬.云南铁皮石斛和齿瓣石斛DALP和ITS遗传变异研究[D].昆明:云南大学,2007.