Screening and Comparison of Reference Genes for microRNA Quantitative Real-time PCR in Jatropha curcas Under Chilling Stress

doi:10.13560/j.cnki.biotech.bull.1985.2019-0042

Abstract

Abstract: Screening appropriate reference gene is of essential importance for the accuracy of quantitative real-time PCR（qRT-PCR）results,and it is especially true in qRT-PCR analysis of microRNAs（miRNAs）. Screening suitable reference genes for miRNA qRT-PCR analysis in Jatropha curcas under chilling stress may provide important reference genes for qRT-PCR analysis of miRNAs in J. curcas and other species. Eleven candidate genes were selected from the chilling-treated J. curcasbased on our previous small RNA-seq results,and their expression levels in different samples were detected by qRT-PCR. The expression stability was analyzed by GeNorm,NormFinder,and BestKeeper software. The results showed that the most stable expression were miR6448 and U6 genes. miR6448 had moderate expression abundance and its Ct was 23. U6 gene had high expression abundance,and the CT was about 10. Therefore,miR6448 can be used as the moderate expression abundance reference gene for miRNA qRT-PCR analysis in J. curcas under chilling stress. U6 gene can be used as the high abundance reference gene for miRNA qRT-PCR analysis in J. curcas under chilling stress.

Key words: Jatropha curcas, quantitative real-time PCR, reference gene, miRNA, chilling stress

KONG Chun-yan, CHEN Yong-kun, WANG Sha-sha, HAO Da-hai, YANG Yu, GONG Ming. Screening and Comparison of Reference Genes for microRNA Quantitative Real-time PCR in Jatropha curcas Under Chilling Stress[J]. Biotechnology Bulletin, 2019, 35(7): 25-32.

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