Selection of Optimized Reference Genes for qRT-PCR Normalization in Xanthomonas campestris pv. campestris Cultured in Different Media

Selection and validation of reference genes for gene expression studies in Xanthomonas arboricola pv. juglandis subjected to abiotic stress

“…out to evaluate RGs in plant-pathogenic bacteria, demonstrating that it is not possible to define universal RGs that are stably expressed under different experimental conditions (Jiang et al, 2019;Yan et al, 2019;Yang et al, 2018).…”

“…Furthermore, gyrB was selected as the best reference gene, followed by gyrA, rpoB, and gapdh, in X. campestris pv. campestris cultured in different media (Yan et al, 2019) and the second most stable RG in Corynebacterium glutamicum under different stress factors (Wang et al, 2018).…”

“…Gene expression analysis is a powerful tool used to characterize the biological pathways networks involved in plant–pathogen interactions and to improve knowledge on how pathogens adapt to stress conditions (Yan et al, 2019). Although several techniques, such as droplet digital PCR, microarray, transcriptome profiling, and multiplex quantitative PCR, have been developed and used with sensitivity and accuracy in gene expression studies, reverse‐transcription quantitative PCR (RT‐qPCR) remains the most widely used technique for the detection of gene expression or the validation of RNA‐sequencing data.…”

“…A few studies have been carried out to evaluate suitable candidate RGs for accurate normalization of RT‐qPCR data in plant‐pathogenic bacteria, such as Xanthomonas campestris pv. campestris (Yan et al, 2019), Pectobacterium atrosepticum (Takle et al, 2007), Erwinia amylovora (Kałużna et al, 2017), Ralstonia pseudosolanacearum (Yang et al, 2018), Clavibacter michiganensis (Jiang et al, 2019), Xanthomonas citri subsp. citri (Jacob et al, 2011), and Xanthomonas fragariae (Kałużna et al, 2019).…”

“…In this study, nine RGs belonging to different functional classes were selected. RNA polymerase β subunit ( rpoB ), DNA recombination protein ( recA ), gyrase subunit B ( gyrB ), and gyrase subunit A ( gyrA ) have been validated as RGs in previous studies in other plant‐pathogenic bacteria (Jiang et al, 2019; Kałużna et al, 2017; Takle et al, 2007; Yan et al, 2019; Yang et al, 2018). Other genes such as RNA polymerase γ subunit ( rpoC ), RNA polymerase sigma α factor ( rpoA ), rho termination factor ( rho ), ATP synthase subunit δ ( atpD ), and RNA polymerase σ factor RpoD ( rpoD ) were used as RGs in Gluconacetobacter diazotrophicus (Galisa et al, 2012) and Rahnella aquatilis (Li et al, 2019).…”