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Background  Members of plant WRKY transcription factor families are widely implicated in defense responses and various other physiological processes. For canola ( Brassica napus  L.), no WRKY genes have been described in detail. Because of the economic importance of this crop, and its evolutionary relationship to  Arabidopsis thaliana , we sought to characterize a subset of canola  WRKY  genes in the context of pathogen and hormone responses.    Results  In this study, we identified 46  WRKY  genes from canola by mining the expressed sequence tag (EST) database and cloned cDNA sequences of 38  BnWRKY s. A phylogenetic tree was constructed using the conserved WRKY domain amino acid sequences, which demonstrated that BnWRKYs can be divided into three major groups. We further compared  BnWRKYs  to the 72  WRKY  genes from  Arabidopsis  and 91  WRKY  from rice, and we identified 46 presumptive orthologs of  AtWRKY  genes. We examined the subcellular localization of four BnWRKY proteins using green fluorescent protein (GFP) and we observed the fluorescent green signals in the nucleus only.  The responses of 16 selected  BnWRKY  genes to two fungal pathogens,  Sclerotinia sclerotiorum  and  Alternaria brassicae , were analyzed by quantitative real time-PCR (qRT-PCR). Transcript abundance of 13  BnWRKY  genes changed significantly following pathogen challenge: transcripts of 10  WRKY s increased in abundance, two  WRKY  transcripts decreased after infection, and one decreased at 12 h post-infection but increased later on (72 h). We also observed that transcript abundance of 13/16  BnWRKY  genes was responsive to one or more hormones, including abscisic acid (ABA), and cytokinin (6-benzylaminopurine, BAP) and the defense signaling molecules jasmonic acid (JA), salicylic acid (SA), and ethylene (ET). We compared these transcript expression patterns to those previously described for presumptive orthologs of these genes in  Arabidopsis  and rice, and observed both similarities and differences in expression patterns.    Conclusion  We identified a set of 13  BnWRKY  genes from among 16  BnWRKY  genes assayed, that are responsive to both fungal pathogens and hormone treatments, suggesting shared signaling mechanisms for these responses. This study suggests that a large number of BnWRKY proteins are involved in the transcriptional regulation of defense-related genes in response to fungal pathogens and hormone stimuli.

bmc plant biology

Identification and expression analysis of WRKY transcription factor genes in canola (Brassica napus L.) in response to fungal pathogens and hormone treatments