基于AFLP分子标记的我国不同生态栽培产区枸杞资源遗传多样性分析  

Genetic diversity analysis of Lycium chinense samples from different cultivation areas based on AFLP molecular markers

作　　者：张芳 陆瑞华 傅焕哲 张文华 李重 任钢 陈占平 赵玉玲 郭盛 钱大玮 段金廒 ZHANG Fang;LU Rui-hua;FU Huan-zhe;ZHANG Wen-hua;LI Zhong;REN Gang;CHEN Zhan-ping;ZHAO Yu-ling;GUO Sheng;QIAN Da-wei;DUAN Jin-ao(School of Pharmacy,Nanjing University of Chinese Medicine,Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization/National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,Nanjing 210023,China;Bairuiyuan Gouqi Co.,Ltd.,Yinchuan 750200,China;Haixi Agricultural Science Research Institute of Mongolian and Tibetan Autonomus Prefecture,Delingha 817000,China;Jinghe Gouqi Industry Development Center of Bortala Mongolian Autonomous Prefecture,Bortala 833399,China)

机构地区：[1]南京中医药大学药学院,江苏省中药资源产业化过程协同创新中心/中药资源产业化与方剂创新药物国家地方联合工程研究中心,江苏南京210023 [2]宁夏百瑞源枸杞股份有限公司,宁夏银川750200 [3]青海省海西蒙古族藏族自治州农业科学研究所,青海德令哈817000 [4]新疆维吾尔自治区博尔塔拉蒙古自治州精河县枸杞产业发展中心,新疆博尔塔拉833399

出　　处：《中草药》2023年第18期6074-6083,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81773837);江苏省中药资源产业化过程协同创新中心重点项目(ZDXM-2020-02,ZDXM-2022-36);国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-D-202005);中央本级重大增减支项目(2060302);宁夏自然科学基金项目(2022AAC03215)。

摘　　要：目的利用荧光标记辅助的扩增片段长度多态性(amplified fragment length polymorphism,AFLP)技术,从基因组DNA水平上分析我国6个栽培生产区域52份枸杞Lycium chinense样本的遗传多样性。方法对提取的枸杞DNA样本进行酶切、连接、预扩增和选扩增等步骤,扩增产物经毛细管电泳系统分离和数据采集后,利用Popgene软件计算遗传多样性参数,利用GenAlex软件计算遗传距离并进行主坐标分析,利用MEGA-X软件根据遗传距离进行聚类分析,利用Structure软件进行群体遗传结构分析。结果共筛选得到10对AFLP选择性扩增引物组合,扩增得到328条扩增片段,其中多态性片段121条,且在不同产地枸杞居群间多态性位点分布不均匀。分析显示不同栽培产区枸杞的遗传多样性较低,等位基因数(observed number of alleles,Na)、有效等位基因数(effective number of alleles,Ne)、观测杂合度(Nei’s gene diversity,H)和香浓信息指数(Shannon information index,I)分别为1.4380、1.2316、0.1407和0.2152,Nei’s遗传相似性范围为0.8354~0.8902。结论不同栽培生产区域的枸杞居群间存在较为频繁的基因交流,遗传分化程度中等。居群内部遗传变异是枸杞遗传变异的主要来源,居群间遗传变异不显著,同时遗传距离与地理距离没有明显相关性。可为枸杞的遗传背景信息积累、引种栽培、种质资源保护、核心种质库构建以及可持续开发策略的制定和管理提供科学依据。Objective The genetic diversity of 52 Lycium chinense samples from six cultivation areas in China was analyzed at the genomic DNA level by using the fluorescence assisted amplified fragment length polymorphism(AFLP)technique.Methods After the successive steps of enzyme digestion,ligation,pre-amplification and selective amplification of the extracted DNA samples,the PCR products were separated by capillary electrophoresis system.The collected data were then analyzed by Popgene software to calculate the genetic diversity parameters,GenAlex software was used to calculate genetic distances and perform principal coordinate analysis,cluster analysis based on genetic distance was performed using MEGA-X software and population genetic structure analysis was performed using Structure software.Results A total of ten pairs of selective amplification primers were screened,and 328 amplified fragments were amplified,including 121 polymorphic fragments,which were unevenly distributed among Lycium populations from different habitats.The genetic diversity of L.chinense in different habitats was relatively low.The allele number Na,effective allele number Ne,observed heterozygosity H,and Shannon information index I were 1.4380,1.2316,0.1407 and 0.2152,respectively.Nei’s genetic consistency ranged from 0.8354 to 0.8902.There were frequent gene exchanges among populations of L.chinense in different cultivation and production areas.Conclusion There were relatively frequent gene exchanges between Lycium populations in different cultivation and production areas,and the degree of genetic differentiation was moderate,and the genetic variation within populations was the main source of genetic variation of L.chinense.Furthermore,no significant correlation between genetic distance and geographical distance was observed.This study will provide scientific basis for the accumulation of genetic background information,introduction and cultivation,protection of germplasm resources,construction of core germplasm bank and formulation and managemen

关 键 词：枸杞属 扩增片段长度多态性 种质资源 遗传多样性 栽培生产区域 

分 类 号：R286.2[医药卫生—中药学]