Role of zinc-finger transcription factor NTT/WIP2 in regulating cambial activity and xylem proliferation

Plant vascular cambium development involves dynamic changes in transcriptional profiles, which are mediated by key transcription factors. Understanding secondary growth through complex molecular processes of vascular cambium development has remarkable significance in maximizing carbon storage in trees and is crucial for the genetic improvement of trees for wood production. Here, we studied the functions of NO TRANSMITTING TRACT (NTT)/WIP DOMAIN PROTEIN2 (WIP2) zinc-finger transcription factor in regulating vascular cambium activity and xylem formation using Arabidopsis roots and tree stems as model organisms. Reporter lines carrying red fluorescent protein driven by the promoter of NTT/WIP2 showed cambium specific expression in Arabidopsis roots under confocal microscope. Inducible overexpression of NTT/WIP2 after cambium activation reduced vascular diameter, primary phloem pole distance and secondary vessel number in Arabidopsis roots. In loss of function ntt mutants, significant reductions of vascular diameter, phloem pole distance and number of secondary vessels were also evident. Our data suggests that, both increased expression and loss of function of NTT/WIP2 gene significantly reduce secondary growth in Arabidopsis roots. Simultaneous to our Arabidopsis experiments we also tested the possibility of increasing wood formation amounts through manipulation of PttNTT (putative Populus ortholog of Arabidopsis NTT/WIP2 gene) expression in hybrid aspen trees. But increased expression of NTT/WIP2 in Poplar significantly reduced plant height, stem girth and internode numbers. Altogether, we hypothesize that, appropriate spatiotemporal expression pattern and levels of NTT/WIP2 is required for cambial activity to proceed properly in plants. These findings represent a new foundation toward a more comprehensive understanding of the molecular mechanisms of NTT/WIP2 gene in regulation of cambium development and secondary growth in plants