Selection of Suitable Internal Reference Genes for the Real-Time Quantitative PCR in Streptomyces rimosus M527 and Evaluation of Their Stabilities

doi:10.16409/j.cnki.2095-039x.2020.03.020

Abstract

Abstract: Streptomyces rimosus M527, a rimocidin producer, has a strong antagonistic effect against a variety of plant pathogenic fungi. In order to analyze the biosynthetic and regulatory mechanism of rimocidin in strain M527 and its resistant mutant at transcriptional level, it is very important to select the most stable internal reference gene for the strain M527 and its resistant mutant. In this study, 16S rRNA_sr, hrdB_sr, rpoA_sr, sigF_sr, sigB_sr, gyrB_sr were selected as candidate reference genes for real-time quantitative PCR, and their stabilities were analyzed in wild-type strain M527 and its resistant strains M527-GR7 (higher-rimocidin producer) and M527-GR21 (lower-rimocidin producer) by using software geNorm, NormFinder, BestKeeper, so as to select the internal reference gene with the highest stability. The results showed that sigB_sr gene was the most stable internal reference gene among all tested internal reference genes either in the wild-type M527 or in resistant mutants M527-GR7 and M527-GR21. Subsequently, the relative expression of the structural gene rimG_sr in strain M527-GR7 was detected at different time during fermentation process by using the genes sigB_sr, 16S rRNA_sr and rpoA_sr as internal reference genes. The results revealed that analysis at transcriptional level could be more accurate when the gene sigB_sr was used as an internal reference gene.

Key words: Streptomyces rimosus M527, real-time quantitative PCR, internal reference genes, stability

CLC Number:

S467

XU Jie, HU Yefeng, LIAO Zhijun, MA Zheng, YU Xiaoping. Selection of Suitable Internal Reference Genes for the Real-Time Quantitative PCR in Streptomyces rimosus M527 and Evaluation of Their Stabilities[J]. Chinese Journal Of Biological Control, 2020, 36(3): 429-436.

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