Screening of reference genes for real-time fluorescence quantitative PCR in Kentucky bluegrass

doi:10.11686/cyxb2016297

Abstract

Abstract: To select the most stable reference genes of Kentucky bluegrass (Poa pratensis) for real-time quantitative PCR, we assessed the mRNA expression in different tissues of six traditional candidate reference genes at various leaf development stages, under abiotic stress, or following various hormone applications. Genes assessed included ACT, GAPDH, UBQ, EF-1α, 18s rRNA and β-tubulin. The expression stabilities of these six reference genes were evaluated by geNorm, NormFinder and BestKeeper. It was found that mRNA expression of the candidate reference genes differed significantly between the tissue types, developmental stages, hormone applications or under abiotic stress. In Kentucky bluegrass leaves, GAPDH is recommended as the reference gene for analyzing the mRNA expression in different tissues; EF-1α was the most stable gene under abiotic stress; β-tubulin was most consistent with different hormones; ACT was stably expressed in the different leaf development stages. In conclusion, use of the most appropriate reference genes under different conditions would enhance the accuracy analysis of gene expression in the Kentucky bluegrass. These findings may be applicable when selecting reference genes for analysis of mRNA expression in other members of the Poa genus.

ZHANG Lan, TAN Peng-Hui, TENG Ke, YAN Meng-Ju, HE Chun-Yan, GAN Lu, YIN Shu-Xia. Screening of reference genes for real-time fluorescence quantitative PCR in Kentucky bluegrass[J]. Acta Prataculturae Sinica, 2017, 26(3): 75-81.

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