建立番木瓜性别研究的cDNA-AFLP分析体系  

Construction of cDNA-AFLP Analysis System for Carica papaya Sex

作　　者：李亚丽[1,2] 沈文涛[1] 言普[1] 熊建平[3] 周鹏[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所农业部热带生物技术重点实验室,海口571101 [2]海南大学,海口570228 [3]茂名学院静电研究所,广东茂名525000

出　　处：《热带作物学报》2009年第4期515-519,共5页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.30760134)(No.30560082);中央级事业单位基本科研业务费(2008)资助

摘　　要：以番木瓜幼苗为材料,利用多重PCR技术快速鉴定其性别,建立适合番木瓜性别研究的cDNA-AFLP分析体系,分析影响cDNA-AFLP分析体系的各种可能因素(RNA提取、酶切与连接效率、预扩增效果等),并对影响选择性扩增反应的各因子进行优化。结果表明,选择性扩增的最佳体系,即在反应体系中,各因素终浓度分别为:Tap酶0.06U/μL;Mg2+2.4mmol/L;dNTP2.0mmol/L;引物0.48μmol/L。通过优化体系得到的选择性扩增产物,银染检测后条带清晰稳定。表明成功建立了适合番木瓜性别研究的cDNA-AFLP分析体系。Three sex types of papaya seedlings used as materials were determined by ways of multiple PRC. The factors influencing cDNA-AFLP (amplified fragment length polymorphism) analysis system were analyzed, such as RNA extraction, double enzymes restriction, adapter ligation and pre-amplification, etc., and an optimal selective amplification reaction system was established by optimizing the influencing factors, and their final concentration in the reaction system was 0.06 U/L for Tapase, 2.4 mmol/L for Mg2+, 2.0 ...

关 键 词：番木瓜 性别 cDNA-AFLP分析体系 选择性扩增体系 

分 类 号：S66[农业科学—果树学]