Native vascular related NAC transcription factors are efficient regulator of multiple classes of secondary wall associated genes in banana.

Secondary-wall deposition in xylem vessel elements is regulated by vascular-related NAC transcription factors (VNDs). We show that three banana VNDs (MusaVND1, MusaVND2 and MusaVND3) directly regulate multiple secondary-wall associated genes by binding to their 5'-upstream regulatory region. Transgenic banana harboring either PMusaVND1:GUS, PMusaVND2:GUS or PMusaVND3:GUS showed specific GUS staining in lignified tissues. MusaVND1, MusaVND2 and MusaVND3 encodes transcriptional-activators as its C-terminal region drive expression of reporter genes in vivo in yeast. Purified MusaVND1, MusaVND2 and MusaVND3 proteins in gel shift assay bind to 19-bp secondary-wall NAC binding element (SNBE) while it fails to bind mutated SNBE. Putative SNBE sites in the 5'-upstream regulatory region of important secondary-wall associated genes related to programmed cell death (XCP1), cell-wall modification (IRX1/CesA8, IRX3/CesA7,IRX5/CesA4, IRX8, IRX10 and IRX12) and transcriptional regulation (MYB52, MYB48/59, MYB85, MYB58/72, MYB46, and MYB83) in banana was identified and mobility of these regulatory regions got retarded by MusaVND1, MusaVND2 and MusaVND3. Transcript level of these important secondary wall associated genes were elevated in transgenic banana overexpressing either MusaVND1, MusaVND2 or MusaVND3. Present study suggested promoters with prospective utilization in wall modification in banana (a potential biofuel crop) and suggest a complex transcriptional regulation of secondary wall deposition in plants.