基于MSAP技术的不同栽培生产区域宁夏枸杞DNA甲基化多态性分析  

DNA methylation diversity analysis of Lycium barbarum samples from different cultivation areas based on MSAP

作　　者：张芳 卢有媛 尚尔鑫 郭盛 鲁学军 任钢 陈占平 赵玉玲 钱大玮 段金廒 ZHANG Fang;LU You-yuan;SHANG Er-xin;GUO Sheng;LU Xue-jun;REN Gang;CHEN Zhan-ping;ZHAO Yu-ling;QIAN Da-wei;DUAN Jin-ao(Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization/National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Key Laboratory of Hui Ethnic Medicine Modernization of Ministry of Education,School of Pharmacy,Ningxia Medical University,Yinchuan 750021,China;Ningxia Gouqi Innovation Center,Yinchuan 750002,China;Haixi Agricultural Science Research Institute of Mongolian and Tibetan Autonomous 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]宁夏医科大学药学院教育部回族医药现代化重点实验室,宁夏银川750021 [3]宁夏枸杞创新中心,宁夏银川750002 [4]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 [5]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：《中国中药杂志》2022年第2期392-402,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81773837)。

摘　　要：宁夏枸杞在我国不同栽培产区环境中,发生了明显的表观遗传变异和分化。为探讨我国不同栽培产区宁夏枸杞DNA甲基化水平和模式的差异及变化规律,该文利用荧光标记辅助的甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)技术,对我国主要栽培产区宁夏银川平原、内蒙古巴彦淖尔、甘肃靖远、甘肃玉门、青海德令哈、新疆精河的53份宁夏枸杞基因组DNA样品进行甲基化水平和多态性分析。该研究建立了适用于宁夏枸杞基因组甲基化分析的MSAP技术体系,共筛选出10对MSAP选择性扩增引物,在所有共扩增出的5′-CCGG-3′甲基化位点中,35.85%为全甲基化位点、39.88%为半甲基化位点,表现出丰富的表观遗传分化程度。化学计量分析显示,影响宁夏枸杞表观遗传特性的主要因素是生态环境因素,其次是栽培品种差异因素,降水量、气温、土壤酸碱度等是影响各产地DNA甲基化差异的主要生态因子。该研究有助于深入理解宁夏枸杞在不同生态环境的适应策略,为枸杞子的种质溯源和道地性评判提供新的研究思路与方法。Obvious epigenetic differentiation occurred on Lycium barbarum in different cultivation areas in China.To investigate the difference and change rule of DNA methylation level and pattern of L.barbarum from different cultivation areas in China,the present study employed fluorescence-assisted methylation-sensitive amplified polymorphism(MSAP)to analyze the methylation level and polymorphism of 53 genomic DNA samples from Yinchuan Plain in Ningxia,Bayannur city in Inner Mongolia,Jingyuan county and Yumen city in Gansu,Delingha city in Qinghai,and Jinghe county in Xinjiang.The MSAP technical system suitable for the methylation analysis of L.barbarum genomic DNA was established and ten pairs of selective primers were selected.Among amplified 5′-CCGG-3′methylated sites,there were 35.85%full-methylated sites and 39.88%hemi-methylated sites,showing a high degree of epigenetic differentiation.Stoichiometric analysis showed that the ecological environment was the main factor affecting the epigenetic characteristics of L.barbarum,followed by cultivated varieties.Precipitation,air temperature,and soil pH were the main ecological factors affecting DNA methylation in different areas.This study provided a theoretical basis for the analysis of the epigenetic mechanism of L.barbarum to adapt to the diffe-rent ecological environments and research ideas for the introduction,cultivation,and germplasm traceability of L.barbarum.

关 键 词：宁夏枸杞 DNA甲基化 甲基化敏感扩增多态性 表观遗传 生态因子 

分 类 号：S567.19[农业科学—中草药栽培]