The R2R3-MYB proteins comprise one of the largest families of transcription factors in plants. Although genome-wide analysis of this family has been carried out in some plant species, little is known about R2R3-MYB genes in canola (Brassica napus L.). In this study, we have identified 76 R2R3-MYB genes in the canola genome through mining of expressed sequence tags (ESTs). The cDNA sequences of 44 MYB genes were successfully cloned. The transcriptional activities of BnaMYB proteins encoded by these genes were assayed in yeast. The subcellular localizations of representative R2R3-MYB proteins were investigated through GFP fusion. Besides, the transcript abundance level analysis during abiotic conditions and ABA treatment identified a group of R2R3-MYB genes that responded to one or more treatments. Furthermore, we identified a previously functionally unknown MYB gene-BnaMYB78, which modulates reactive oxygen species (ROS)-dependent cell death in Nicotiana benthamiana, through regulating the transcription of a few ROS- and defence-related genes. Taken together, this study has provided a solid foundation for understanding the roles and regulatory mechanism of canola R2R3-MYB genes.
The NAC (NAM, ATAF1/2, CUC2) transcription factor gene family is plant-specific and plays diverse roles in development and responses to abiotic stresses and pathogen challenge. Oilseed rape (Brassica napus) or canola is an important oil crop worldwide, however, the function of NAC genes in it remains largely elusive. In the present study, we identified and characterized the NAC56 gene isolated from oilseed rape. Expression of BnaNAC56 was induced by abscisic acid (ABA), jasmonic acid (JA), methyl viologen (MV) and a necrotrophic fungal pathogen Sclerotinia sclerotiorum, but repressed by cold. BnaNAC56 is a transcription activator and localized to nuclei. Overexpression of BnaNAC56 induced reactive oxygen species (ROS) accumulation and hypersensitive response (HR)-like cell death, with various physiological measurements supporting these. Furthermore, BnaNAC56 expression caused evident nuclear DNA fragmentation. Moreover, quantitative reverse transcription PCR (qRT-PCR) analysis identified that the expression levels of multiple genes regulating ROS homeostasis, cell death and defense response were significantly induced. Using a dual luciferase reporter assay, we further confirmed that BnaNAC56 could activate the expression of a few ROS- and cell death-related genes. In summary, our data demonstrate that BnaNAC56 functions as a stress-responsive transcriptional activator and plays a role in modulating ROS accumulation and cell death.
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