Comprehensive Selection of Reference Genes for Gene Expression Normalization in Sugarcane by Real Time Quantitative RT-PCR
The increasingly used real time quantitative reverse transcription-PCR (qRT-PCR) method for gene expression analysis requires one or several reference gene(s) acting as normalization factor(s).In order to facilitate gene expression studies in sugarcane (50ccharum officinorum),a non-model plant with limited genome information,the stability of 13 candidate reference genes was evaluated.The geNorm,NormFinder and deltaCt methods were used for selecting stably expressed internal controls across different tissues and under various experimental treatments.These results revealed that,among these 13 candidate reference genes,GAPDH,eEF-1a and e/F-4α were the most stable and suitable for use as normalization factors across all various experimental samples.In addition,APRT could be a candidate for examining the relationship between gene copy number and transcript levels in sugarcane tissue samples.According to the results evaluated by geNorm,combining CUL and eEF-1α in hormone treatment experiments; CAC and CUL in abiotic stress tests; GAPDH,eEF-1α and CUL in all treatment samples plus CAC,CUL,APRT and TIP5-41 in cultivar tissues as groups for normalization would lead to more accurate and reliable expression quantification in sugarcane.This is the first systematic validation of reference genes for quantification of transcript expression profiles in sugarcane.This study should provide useful information for selecting reference genes for more accurate quantification of gene expression in sugarcane and other plant species.
Hui Ling Qibin Wu Jinlong Guo Liping Xu Youxiong Que
Key Laboratory of Sugarcane Biology and Genetic Breeding,Ministry of Agriculture,Fujian Agriculture and Forestry University,Fuzhou,Fujian,China
国内会议
海口
英文
54-63
2014-08-01（万方平台首次上网日期，不代表论文的发表时间）