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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。
Real-time quantitative PCR technology is widely used in the study of plant functional gene transcription level changes. Choosing the appropriate internal control gene for relative quantitative analysis is the key factor for accurate experimental results. The results showed that the eEF-1α gene was expressed in rice after mechanical damage treatment by analyzing the stability of the expression of five commonly used internal control genes (eEF-1α, 18SrRNA, 25SrRNA, Actin and UBQ5) in rice (Oryza sativa) The expression of Actin gene was the most stable after treatment of rice stem borer (Chilo suppressalis), and the expression of Actin gene was the most stable after treated with rice stem borer (Chilo suppressalis) The most stable expression. At the same time, these internal control genes were evaluated using the expression of OsHI-LOX gene after different treatments. The results provide a theoretical basis for the selection of internal control genes in real-time quantitative PCR analysis of rice pests.