Open Access

ARTICLE

Selection and validation of reference genes for quantitative real-time polymerase chain reaction analyses of Serratia ureilytica DW2

FENGLIN BAI^1,2,#, BIANXIA BAI^1,2,#, TINGTING JIN^1,2, GUIPING ZHANG^1,2, JIAHONG REN^1,2,*

1 The Department of Life Sciences, Changzhi University, Changzhi, China
2 Ecological and Environmental Research Institute of Taihang Mountain, Changzhi, China

* Corresponding Author: Jiahong Ren, # These authors contributed equally to this work.

(This article belongs to this Special Issue: Advances in Plant Cell Biology Research)

BIOCELL 2023, 47(3), 647-656. https://doi.org/10.32604/biocell.2023.024758

Received 27 July 2022; Accepted 07 September 2022; Issue published 03 January 2023

Abstract

Background: Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C. pilosula; nonetheless, until now, no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction (RT–qPCR) data. Methods: To screen stable reference genes of S. ureilytica DW2, the expression of its eight candidate reference genes (16S rRNA, ftsZ, ftsA, mreB, recA, slyD, thiC, and zipA) under different treatment conditions (pH, temperature, culture time, and salt content) was assayed by RT–qPCR. The expression stability of these genes was analyzed using different algorithms (geNorm, NormFinder, and BestKeeper). To verify the reliability of the data, the expression of the glucose dehydrogenase (gdh) gene under different soluble phosphate levels was quantified using the most stably expressed reference gene. Results: The results showed that the zipA and 16S rRNA genes were the most stable reference genes, and the least stable genes were thiC and recA. The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium. Conclusion: Therefore, this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.

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