香菇反转录转座子间扩增多态性(IRAP)PCR反应体系的研究  

Study on PCR Reaction System of Inter-retrotransposon Amplified Polymorphism(IRAP) in Lentinula edodes

作　　者：肖扬[1] 李黎[1] 刘伟[1] 边银丙[1] 

机构地区：[1]华中农业大学应用真菌研究所,武汉430070

出　　处：《中国农学通报》2009年第7期47-51,共5页Chinese Agricultural Science Bulletin

基　　金：农业部公益性行业(农业)科研专项项目"食用菌菌种质量评价与菌种信息系统研究与建立"(nyhyzx07-008);农业部948项目"食用菌优异种质与优质安全生产技术引进与创新"(2006-G11(3)-7)。

摘　　要：为了开发香菇反转录转座子间扩增多态性标记(IRAP),建立稳定的IRAP-PCR反应体系,对影响IRAP-PCR的主要因素进行了优化筛选。确定了最佳反应体系为:20μl反应体系中,包含30ng模板DNA,0.30μmol/L引物,0.3mmol/L dNTPs,2.0mmol/L Mg2+及0.75UTaq酶。梯度PCR试验筛选得到的最佳退火温度为56.1℃。采用上述最佳反应体系和引物LTR1L-MarY1L对香菇18个菌株进行了IRAP-PCR扩增,验证了该体系的可靠性。The main factors affecting IRAP-PCR amplification were optimized in order to develop inter-retrotransposon amplified polymorphism marker and to establish stable IRAP reaction system in Lentinula edodes. The optimal IRAP-PCR system was as follows, within 20μl reaction system, there were 30 ng template DNA, 0.3 μmol/L primer, 0.3mmol/L dNTPs, 2.0 mmol/L Mg^2＋ and 0.75 U Taq DNA polymerase. The optimal annealing temperature was determined as 56.1℃ by gradient PCR. Based on the optimal IRAP-PCR system, primer combination LTR1L and MarY1L was used to amplify genomie DNA of eighteen strains. Results indicated that the optimal IRAP-PCR system is reliable.

关 键 词：香菇 反转录转座子 IRAP-PCR 体系优化 

分 类 号：S646.12[农业科学—蔬菜学]