云南农业大学学报2011，Vol.26Issue(5)：602-606,5.DOI:10.3969/j.issn.1004-390X(n).2011.05.004
烟草赤星病菌ISSR-PCR反应体系的优化
Optimization of ISSR-PCR Reaction System for Alternaria alternate in Tobacco
摘要
Abstract
The genomic DNA was extracted from Alternaria alternata mycelium infecting Tobacco using modified CTAB method. Using the genomic DNA of A. Alternate, the factors influencing ISSR-PCR system were optimized with the orthogonal and single factor variable design. The results show that the system, which contains 2. 5 μL 10 ℃ buffer, 20ng template, 2. 0 mmol/L Mg2+ , 0. 8 μmol/L primer , 0. 2mmol/L dNTPs, 0. 75 U Taq DNA polymerase in 25 μL reaction is suitable for A. Alternata ISSR molecular marker analysis, DNA of eight samples collected from Kunming, Dehong, Chuxiong, Qujing, Yuxi and other areas were used in ISSR-PCR application with this system and results indicate it is suitable for this reaction system.关键词
烟草赤星病/ISSR-PCR/正交体系优化Key words
Alternaria alternate/ ISSR-PCR/optimization of rthogonal分类
植物保护学引用本文复制引用
胡中会,赵立华,佟爱仔,王连春,孔宝华,蔡红,范静华,陈海如,秦西云..烟草赤星病菌ISSR-PCR反应体系的优化[J].云南农业大学学报,2011,26(5):602-606,5.基金项目
中烟公司科技计划项目(09Y N005) （09Y N005）
中烟公司科技计划项目(2010YN18) （2010YN18）
中烟公司科技计划项目(2010YN19). （2010YN19）
