江西农业大学学报2012，Vol.34Issue(6)：1086-1092,7.
开花期水稻颖花实时定量RT-PCR分析中内参基因的选择
Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Rice Florets during Anthesis
摘要
Abstract
Quantitative real - time reverse transcription - polymerase chain reaction ( qRT-PCR) has become the most common method for gene expression analysis. In order to get accurate and reliable qRT-PCR results , stably expressed reference genes are required for normalization of tested samples. The expression stabilities of 6 commonly used reference genes (ACT1 , eEFl-α, β - TUB, UBQ5 , GAPDH and UBC) were carefully assessed by geNorm algorithm in florets during anthesis in japonica rice ' Wuyunjing 7 ' and indica rice 'Minghui 63'. The results showed that the expression of eEFl -α and β - TUB was the most stable in the floret samples of ' Wuyunjing 7 '. UBC and GAPDH exhibited the most stable expression in ' Minghui 63 '. In contrast, ACT1 was found to be the least stable expression among the tested samples from both rice varieties. Moreover , a combination of two most stable genes was found to be better as internal control for normalization of the data. The effect of normalization with different internal controls on quantification of OsAOC, a key gene in JA biosynthesis, was also examined. The relative quantification of expression of OsAOC varied according to the internal controls used, thus highlighting that choice of stable reference genes for normalization was a critical precondition for reliable qRT-PCR data.关键词
水稻颖花/基因表达/实时定量RT-PCR/内参基因Key words
rice florets/ gene expression/ quantitative real-time RT-PCR/ reference genes分类
农业科技引用本文复制引用
何永明,曾晓春..开花期水稻颖花实时定量RT-PCR分析中内参基因的选择[J].江西农业大学学报,2012,34(6):1086-1092,7.基金项目
国家自然科学基金项目(30960180)和江西省科技支撑计划项目(20111BBF60009) （30960180）
