高粱抗丝黑穗病基因SRAP体系优化  

Optimization SRAP Reaction System of The Head Smut Resistance Gene on Sorghum

作　　者：段永红[1] 渠云芳[1] 王铭[2] 王长彪[2] 司浩浩 张旭[1] 孙毅[2,3] Duan Yonghong Qu Yunfang Wang Ming Wang Changbiao Si Haohao Zhang Xu Sun Yi(College of Agronomy, Shanxi Agricultural University, Taigu, 030801 Biotechnology Research Center, Shanxi Academy of Agricultural Sciences, Taiyuan, 030031 Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Taiyuan, 030031)

机构地区：[1]山西农业大学农学院,太谷030801 [2]山西省农业科学院生物技术研究中心,太原030031 [3]农业部黄土高原作物基因资源与种质创制重点实验室,太原030031

出　　处：《分子植物育种》2017年第2期611-617,共7页Molecular Plant Breeding

基　　金：山西省科技攻关项目(20140311003-4);山西省科技攻关项目(20130311004-3);山西省财政支农项目(2015TGSF-10);国家公益性行业(农业)科研专项(201503121-07)共同资助

摘　　要：为建立高粱抗丝黑穗病基因最佳的SRAP-PCR反应体系,进一步筛选与抗病基因相关的SRAP标记。本研究采用单因素与正交设计相结合的方法,对影响高粱SRAP-PCR体系的5个因素Taq酶、Mg2+、模板DNA、d NTPs和引物进行优化,以期筛选出最优的高粱抗丝黑穗病基因SRAP-PCR反应体系。研究表明:在优化得到的20μL的高粱SRAP-PCR体系中,模板DNA的用量为20.0 ng,Taq DNA聚合酶的用量为0.14 U,Mg2+的浓度为3.0 mmol/L,d NTPs浓度为0.3 mmol/L,引物的浓度为0.5μmol/L。各因素对反应体系影响大小依次为:引物浓度>DNA用量>Taq DNA聚合酶浓度>Mg2+浓度=d NTPs浓度。本研究将为高粱抗性基因的定位与功能研究提供基础数据与技术支持。The purpose of this study was to determine the best SRAP-PCR reaction system of the head smut resistance gene on Sorghum and provide a foundation for further screening SRAP markers of resistance gene.Combining the single factor with the orthogonal experimental design, five factors including the concentrations of template DNA, Mg2＋, d NTPs, Taq DNA polymerase and primers in SRAP-PCR reaction were optimized, then the optimal SRAP-PCR reaction system of the head smut resistance gene on sorghum was established. The results showed that the best SRAP-PCR amplification system for sorghum was 20 ng DNA template, 0.14 U polymerase,3.0 mmol/L Mg2＋, 0.3 mmol/L d NTPs and 0.5 μmol/L primer in a 20 μL reaction system. The order of each factor affecting the result of PCR was： primer〉 DNA 〉DNA Taq polymerase 〉d NTPs =Mg2＋. This study will provide basic data and technical support for the location and function of sorghum resistance genes.

关 键 词：高粱 SRAP-PCR 体系优化 正交设计 

分 类 号：S435.14[农业科学—农业昆虫与害虫防治] Q943.2[农业科学—植物保护]