Establishment and Optimization of ISSR Reaction System for Pea

doi:10.11733/j.issn.1007-0435.2012.03.024

Acta Agrestia Sinica ›› 2012, Vol. 20 ›› Issue (3): 536-539.DOI: 10.11733/j.issn.1007-0435.2012.03.024

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Establishment and Optimization of ISSR Reaction System for Pea

ZENG Liang¹, LI Min-quan^1,2, YANG Xiao-ming², ZHAO Li-juan², YANG Fa-rong²

1. College of Grassland Science, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China;
2. Academy of Gansu Agriculture Science, Lanzhou, Gansu Province 730070, China

Received:2012-02-28 Revised:2012-04-05 Online:2012-06-15 Published:2012-07-05

豌豆ISSR-PCR反应体系的建立和优化

曾亮¹, 李敏权^1,2, 杨晓明², 赵丽娟², 杨发荣²

1. 甘肃农业大学草业学院, 兰州 730070;
2. 甘肃省农业科学院, 兰州 730070

通讯作者: 李敏权,E-mail:lmq@gsau.edu.cn
作者简介:曾亮 (1983- ),男,甘肃兰州人,博士研究生,研究方向为植物病理,E-mail: steve-1314@163.com
基金资助:
现代农业食用豆产业技术体系专项(CARS-09);国家自然科学基金(31160304)资助

Abstract

Abstract: The optimum ISSR-PCR system for peas is established using ISSR-PCR amplification with five factors (DNA template, primer, dNTPs, Taq DNA polymerase and annealing temperature). The optimum reaction system (20 μL) contains 15 ng DNA template, 0.15 mmol·L^-1 dNTP, 0.5 μmol·L^-1 ISSR primer, 1U Taq DNA polymerase. The suitable annealing temperature of primer 842 is 50℃. This reaction system will provide a technological base for genetic diversity analysis of pea germplasm, genetic map construction and gene location.

Key words: Pisum sativum L., ISSR, PCR reaction system, Optimization

摘要： 以豌豆(Pisum sativum L.)DNA为材料,对影响其ISSR-PCR反应体系中的5种主要因素(模板DNA量、引物浓度、dNTPs浓度、Taq酶量、退火温度)进行优化,最终确定了豌豆ISSR-PCR反应的最佳体系(20 μL)为:15 ng模板DNA, 0.15 mmol·L^-1dNTP, 0.5 μmol·L^-1ISSR引物, 1 U Taq DNA聚合酶,引物842号的最适退火温度为50℃。该体系的建立为今后利用ISSR技术进行豌豆属种质资源的遗传多样性分析、遗传图谱构建和基因定位奠定了技术基础。

关键词: 豌豆, ISSR, PCR反应体系, 优化

CLC Number:

Q943

ZENG Liang, LI Min-quan, YANG Xiao-ming, ZHAO Li-juan, YANG Fa-rong. Establishment and Optimization of ISSR Reaction System for Pea[J]. Acta Agrestia Sinica, 2012, 20(3): 536-539.

曾亮, 李敏权, 杨晓明, 赵丽娟, 杨发荣. 豌豆ISSR-PCR反应体系的建立和优化[J]. 草地学报, 2012, 20(3): 536-539.

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Establishment and Optimization of ISSR Reaction System for Pea

豌豆ISSR-PCR反应体系的建立和优化

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