Analysis of the NAC transcription factor gene family in citrus reveals a novel member involved in multiple abiotic stress responses

The NAC (NAM, ATAF1, -2, and CUC2) gene family encodes a large family of plant-specific transcription factors that play diverse roles in plant development and stress regulation. In this study, we performed a survey of citrus NAC transcription factors in the HarvEST: Citrus database, in which 45 NAC domain-containing proteins were identified and phylogenetically classified into 13 different subfamilies. The results suggest the existence of a structurally diversified family of NAC transcription factors in citrus, which has not been previously characterized. One of these NAC genes, CsNAC1 was found to be a member of the stress-NAC subfamily, whose homologs from other plant species function in pathways of environmental stress response and tolerance, and was further characterized. The CsNAC1 deduced protein was shown to contain the five N-terminal A through E NAC subdomains, a C-terminal region containing three transcriptional activation motifs, and a predicted NAC nuclear localization signal, consistent with its putative role as a NAC transcription factor. In silico analysis indicated that CsNAC1 was primarily expressed in leaves and shoot meristems, and was involved in general stress responses. Quantitative real-time reverse transcription PCR analysis revealed that CsNAC1 was strongly induced by drought stress in leaves of Citrus reshni and Citrus limonia, and also by salt stress, cold, and ABA in leaves and roots of C. reshni. Collectively, these results suggest that CsNAC1 encodes a novel stress-responsive NAC transcription factor that is potentially useful for engineering tolerance to multiple abiotic stresses in citrus.