| 油料植物光皮树SSR-PCR反应体系优化及引物筛选 |
| |
| 引用本文： | 张路红,蒋丽娟,陈景震,易诗明,黄蓉,夏栗.油料植物光皮树SSR-PCR反应体系优化及引物筛选[J].经济林研究,2017,35(2). |
| |
| 作者姓名： | 张路红 蒋丽娟 陈景震 易诗明 黄蓉 夏栗 |
| |
| 作者单位： | 1. 中南林业科技大学,湖南长沙,410004;2. 湖南省林业科学院,湖南长沙,410004;3. 湖南省农林工业勘察设计研究总院,湖南长沙,410007 |
| |
| 基金项目： | “十二·五”国家科技支撑计划课题“新型油料能源植物选育及栽培示范”,湖南省研究生科技创新基金项目;中南林业科技大学研究生科技创新基金项目 |
| |
| 摘 要： | 建立稳定可靠的SSR-PCR反应体系、筛选具有多态性SSR引物,是木本油料植物光皮树分子标记辅助育种与良种选育的重要基础研究。为给SSR标记技术在光皮树遗传图谱构建、亲缘关系鉴定及分子育种等方面的应用研究奠定基础,利用"Cf-05"正反向引物,采用L16(45)正交试验设计和单因素试验相结合的研究方法,对影响光皮树SSR-PCR反应体系的主要因素(模板DNA、引物、d NTPs、Mg~(2+)、Taq聚合酶浓度及退火温度)进行了试验分析。试验结果表明,光皮树SSR-PCR总体积为15μL的最佳反应体系为:模板DNA用量37.5 ng,正反向引物浓度均为0.6μmol·L~(-1),d NTPs浓度为0.3 mmol·L~(-1),Mg~(2+)的浓度为1.5 mmol·L~(-1),Taq聚合酶用量为1.5 U,最佳退火温度为50℃。运用优化了的光皮树SSR-PCR反应体系,可从71对引物中筛选获得11对能适用于光皮树的SSR引物。研究结果表明,试验所建立的反应体系可进一步用于光皮树SSR遗传多样性分析、遗传图谱构建和分子标记辅助育种等方面的研究中。
|
| 关 键 词： | 光皮树 SSR-PCR 引物筛选 正交优化试验 |
Optimization and primer screening of SSR-PCR reaction system for Swida wilsoniana |
| |
| ZHANG Luhong,JIANG Lijuan,CHEN Jingzhen,YI Shiming,HUANG Rong,XIA Li.Optimization and primer screening of SSR-PCR reaction system for Swida wilsoniana[J].Economic Forest Researches,2017,35(2). |
| |
| Authors: | ZHANG Luhong JIANG Lijuan CHEN Jingzhen YI Shiming HUANG Rong XIA Li |
| |
| Abstract: | The important basis of the researches on SSR marker assisting selective breeding of woody oil plant Swida wilsoniana is to establish an SSR-PCR reaction system and get some polymorphism primers.In order to lay a foundation for the researches on applying SSR marker technology to establishment of genetic maps,genetic relationship identification,and molecular breeding,some main impact factors in the S.wilsoniana SSR-PCR reaction system were analyzed by using combination method of L16(45) orthogonal test and single factor tests,including DNA templates,primer,dNTPs,Mg2+,Taq polymerase,and annealing temperature,using "Cf-05" forward and reverse primers.The results showed that the optimized PCR reaction mixture was 15 μtL total volume including 37.5 ng template DNA,0.6 μmol/L forward and reverse primers,0.3 mmol/L dNTPs,1.5 mmol/L Mg2+ and 1.5 U Taq DNA polymerase,and the optimum annealing temperature was 50 ℃C.11 pairs of SSR primers suitable for S.wilsoniana could be obtained from 71 pairs primers,through applying the optimized S.wilsoniana SSR-PCR reaction system.This result indicated that the optimized SSR-PCR reaction system could provide a vital theoretic and technical support for genetic diversity analysis,genetic maps establishment and molecular marker assisted breeding in S.wilsoniana. |
| |
| Keywords: | S wilsoniana SSR-PCR primer screening orthogonal optimization test |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
