水稻虫害诱导相关基因实时定量PCR中内参基因的选择  

Validation of Rice Candidate Reference Genes for Herbivore-induced Quantitative Real-time PCR Analysis

作　　者：李冉[1] 李建彩[1] 周国鑫[1] 娄永根[1] 

机构地区：[1]浙江大学昆虫科学研究所,杭州310029

出　　处：《植物学报》2013年第2期184-191,共8页Chinese Bulletin of Botany

基　　金：国家重点基础研究发展计划(No.2010CB126200);国家自然科学基金(No.31101451)

摘　　要：实时定量PCR技术广泛应用于植物功能基因转录水平变化的研究,选择合适的内参基因进行相对定量分析是实验结果准确的关键因素。通过分析5个常用的内参基因(eEF-1α、18SrRNA、25SrRNA、Actin和UBQ5)在水稻(Oryza sativa)经过各种处理后表达的稳定性,结果表明,水稻经过机械损伤处理后eEF-1α基因的表达最稳定;二化螟处理后25SrRNA基因的表达最为稳定;稻纵卷叶螟处理后Actin基因的表达最稳定;两种刺吸式口器昆虫褐飞虱和白背飞虱危害后,UBQ5基因的表达最稳定。同时,利用OsHI-LOX基因在不同处理后的表达来评价这些内参基因。研究结果为水稻虫害诱导实时定量PCR分析中内参基因的选择提供了理论依据。Quantitative real-time PCR is popularly used to study gene transcription levels in plants. Validation of candidate reference genes is the key step to ensure relative quantification. Here, we selected 5 reference genes （eEF-1α, 18S rRNA, 25S rRNA, Actin, and UBQ5） and tested their expression stability in rice under various treatments. eEF-1α showed the most stable expression with mechanical wounding, 25S rRNA with stripped stem borer treatment, and Actin with leaf folder treatment. UBQ5 showed the most stable expression on treatment with 2 phloem-feeding herbivores, brown plant hopper and white-backed plant hopper. We validated the expression profiles of these candidate reference genes by studying that of OsHI-LOX. We offer an available reference for studying gene transcription levels with herbivore infestation in rice.

关 键 词：虫害诱导 实时定量PCR 内参基因 水稻 

分 类 号：S435.112[农业科学—农业昆虫与害虫防治]