摘要
为确立桉树SRAP-PCR反应体系,并对桉树品种进行遗传多样性分析,以巨尾桉GL-9号嫩叶提取的DNA为模板,进行Mg2+、dNTP、引物和Taq DNA聚合酶4个因素3个水平L9(34)正交试验,并比较了不同浓度模板DNA对PCR扩增效果的影响。结果显示:桉树的SRAP-PCR最佳反应体系为Mg2+2.5 mmol/L、dNTP 0.20mmol/L、引物0.4μmol/L、Taq DNA聚合酶1.5U,DNA模板最佳浓度为10ng。利用最佳反应体系对桉树品种进行引物组合多态性筛选,从40个引物组合中筛选出多态性引物组合17个。挑选12个多态较高的引物组合对11个桉树品种进行遗传多样性分析,通过PCR扩增,得到109个谱带。其中多态性条带95条,平均每个引物组合产生7.92个多态性条带,显示了相对较高的多态性。表明SRAP标记可应用于桉树分子生物学研究。
The orthogonal design was used to optimize a SRAP-PCR system with 4 factors(Mg2+,dNTP,primer and Taq polymerase) at 3 levels.The effect of the concentration of template DNA on PCR amplification was compared.The optimized SRAP-PCR system in Eucalyptus was 10ng template DNA,2.5 mmol/L Mg2+,0.2mmol/L dNTP,0.4μmol/L primer,and 1.5U Taq polymerase in a total of 10μL reaction solution.The selected twelve primer pairs out of forty were polymorphic by the optimized SRAP-PCR.Twelve primer pairs produced 109 bands among 11 accessions,showing some genetic diversity in Eucalyptus.The result indicated that SRAP molecular marker could be widely used in molecular research in Eucalyptus.
出处
《林业科技开发》
2011年第6期24-27,共4页
China Forestry Science and Technology
基金
广西林科院基金项目资助(编号:林科201116号)
关键词
桉树
SRAP标记
优化
遗传多样性
Eucalyptus
SRAP marker
optimization
genetic diversity
