PT  - JOURNAL ARTICLE
AU  - Fenglin Bai
AU  - Bianxia Bai
AU  - Tingting Jin
AU  - Guiping Zhang
AU  - Jiahong Ren
TI  - Selection and validation of reference genes for quantitative real-time polymerase chain reaction in <em>Serratia ureilytica</em> DW2
AID  - 10.1101/2022.05.03.490547
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.05.03.490547
4099  - http://biorxiv.org/content/early/2022/05/03/2022.05.03.490547.short
4100  - http://biorxiv.org/content/early/2022/05/03/2022.05.03.490547.full
AB  - Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacterium isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C. pilosula. However, no validated reference genes from the genus Serratia for use in quantitative real-time polymerase chain reaction (RT–qPCR) normalization have been reported. To screen stable reference genes in S. ureilytica DW2, the expression of 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 with different algorithms (geNorm, NormFinder, and BestKeeper). To verify the reliability of the data, the most stably expressed reference gene was used to quantify expression of the glucose dehydrogenase (gdh) gene under different soluble phosphate levels. The results showed that the zipA and 16S rRNA genes were the most stable reference genes, and the least stable were thiC and recA. The expression of gdh was consistent with the phosphate solubilization ability on plates containing National Botanical Research Institute phosphate (NBRIP) growth medium. Therefore, this study provides a stable and reliable reference gene for Serratia, which is vital for the accurate quantification of functional gene expression in future studies.Competing Interest StatementThe authors have declared no competing interest.