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Pseudomonas brassicacearum  GS20 is an antagonistic strain of bacteria recently isolated from the rhizosphere of  Codonopsis pilosula . No validated reference gene has been indentified from  P .  brassicacearum  to use in the normalization of real-time quantitative reverse transcription-PCR data. Therefore, in this study, nine candidate reference genes ( recA ,  gyrA ,  rpoD ,  proC ,  gmk ,  rho ,  16S ,  ftsz , and  secA ) were assessed at different growth phases and under various growth conditions. The expression stability of these candidate genes was evaluated using BestKeeper, NormFinder and GeNorm. In general, the results showed  rho ,  rpoD  and  gyrA  were the most suitable reference genes for  P .  brassicacearum  GS20. The relative expression of iron-regulated gene ( fhu ) was normalized to verify the reliability of the proposed reference genes under iron-replete and iron-limited conditions. The trend in relative expression was consistent with the change in siderophore production under different iron conditions. This study presents reliable reference genes for transcriptional studies in  P .  brassicacearum  GS20 under the chosen experimental conditions.

plos one

Selection and validation of reference genes for gene expression studies in Pseudomonas brassicacearum GS20 using real-time quantitative reverse transcription PCR