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

作　　者：FENGLIN BAI BIANXIA BAI TINGTING JIN GUIPING ZHANG JIAHONG REN 

机构地区：[1]The Department of Life Sciences,Changzhi University,Changzhi,China [2]Ecological and Environmental Research Institute of Taihang Mountain,Changzhi,China

出　　处：《BIOCELL》2023年第3期647-656,共10页生物细胞（英文）

基　　金：supported by the General Program of Natural Science Foundation of China(32071770);the Fundamental Research Program of Shanxi Province(Award No.202103021223380);the Fund for Shanxi“1331 Project”Key Subjects Construction(1331KSC).

摘　　要：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.

关 键 词：Medical plants Codonopsis pilosula Phosphate-solubilizing bacterium Growth-promoting bacteria 

分 类 号：R282.71[医药卫生—中药学]