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、18S rRNA、25S rRNA、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25S rRNA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.