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| Selection of Reference Genes for Real-time Quantitative RT-PCR in Rice(Oryza sativa L. ssp. japonica) under Nutrient Deficiency |
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Abstract Mineral nutrient assimilation and utilization play a critical role in rice yield. Reference genes suitable for nutrient deficiency must be selected, prior to qRT-PCR analysis of rice functional gene. In this study, a total of 12 rice housekeeping genes were selected as candidate reference genes for qRT-PCR of rice under nutrient deficiency. Total sample pool consisted of rice(Oryza sativa L. ssp. japonica) plants grown under 1-week and 2-week nutrient deficiency of 6 macro elements, including nitrogen, phosphorus, potassium, calcium, magnesium, sulphur. Stability of candidate reference genes under nutrient deficiency was analysed and reference gene eIF4a was identified as the most suitable gene using geNorm, and two genes with most stable expression were found to be optimal for reliable normalisation, based on the result of actual normalisation effect under different nutrient deficiency using various reference gene groups, reference gene selection was been proved to be of importance on qRT-PCR results under nutrient deficiency treatment of rice, and effectiveness of gene stability analysis using geNorm was further confirmed. Finally, single gene normalisation of eIF4a was validated as an improvement and complement for geNorm software. Taken together, all the results provide good reference for further research in qRT-PCR under nutrient deficiency.
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Received: 15 August 2013
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