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

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.