The invention discloses screening and application of
burkholderia gladioli real-time quantitative
polymerase chain reaction (qRT-PCR)
reference genes and primers thereof. According to obtained
transcriptome analysis results and reported
reference genes, 12 candidate
reference genes with relatively stable expression are preliminarily screened, and
gene names are shown as follows: atpD, clpP, clpX,
ftsZ, gyrB, lpxC, pyrG, recA,
rpoB , rpoD, thyA and 16S; for the candidate genes, amplification primers are designed; reference genes under different culture conditions (temperature, initial pH valueof medium, culture time, treatment with NaCl different in concentration) of
burkholderia gladioli are screened; geNorm, NormFinder and Bestkeeper
software is employed to analyze qRT-PCR results; and finally, the reference genes with most stable expression under different culture conditions and NaCl treatment condition are obtained by screening. The invention is beneficial to the stability and reliability of analysis and study of
gene expression of the
burkholderia gladioli under different conditions.