山豆根ISSR-PCR反应体系优化及引物筛选  

Optimization of ISSR-PCR Reaction System and Primer Selection of Sophora tonkinensis Gagnep

作　　者：李金梅 关妙娟 黄鼎 明如宏 李良波 谭勇 LI Jin-mei;GUAN Miao-juan;HUANG Ding(College of Pharmacy,Guangxi University of Chinese Medicine,Nanning,Guangxi 530200;Key Laboratory of Resources Protection and Utilization of Traditional Chinese Medicine and Ethnic Medicine,Guangxi University of Chinese Medicine,Nanning,Guangxi 530200)

机构地区：[1]广西中医药大学药学院,广西南宁530200 [2]广西中医药大学/广西高校中药民族药资源保护与利用重点实验室,广西南宁530200

出　　处：《安徽农业科学》2024年第18期78-82,共5页Journal of Anhui Agricultural Sciences

基　　金：广西中医药大学中药学一流学科项目(2019XK093);广西中医药大学青年创新研究团队项目(2018QT001)。

摘　　要：以山豆根为试验材料,采用正交试验考察DNA、引物、Mg^(2+)、dNTPs、Taq酶5个因素对ISSR-PCR反应体系的影响,建立并优化山豆根ISSR-PCR反应体系条件。结果表明:山豆根ISSR-PCR反应体系(20μL)最优条件为Mg^(2+)浓度3 mmol/L,dNTPs 0.2 mmol/L,Taq酶1 U,引物0.5μmol/L,模板DNA 100 ng,运用该体系从100条UBC引物中筛选到10条清晰、多态性好的引物。采用正交试验设计可有效建立山豆根ISSR-PCR反应的最优体系,为山豆根的亲缘关系,种质资源遗传多样性评价及品种鉴定和系统分类等研究提供科学依据。Taking Sophora tonkinensis as materaial,orthogonal experiments were used to investigate the effects of DNA,primers,Mg^(2+),dNTPs,and Taq enzymes on the ISSR-PCR reaction system,and the conditions of ISSR-PCR reaction system of Sophora tonkinensis were established and optimized.The results showed that the optimal conditions for the ISSR-PCR reaction system(20μL)of Sophora tonkinensis including 3 mmol/L Mg^(2+),0.2 mmol/L dNTPs,1 U Taq enzyme,0.5μmol/L primer,100 ng template DNA.Ten primers with good polymorphism were selected from 100 UBC primers by this system.The optimal system of ISSR-PCR reaction can be effectively established by orthogonal experiment design,which provides a scientific basis for the research of genetic relationship,genetic diversity evaluation of germplasm resources,variety identification and systematic classification of Sophora tonkinensis.

关 键 词：山豆根 ISSR-PCR体系优化 正交试验 引物筛选 

分 类 号：S567[农业科学—中草药栽培]