Proceedings of the National Academy of Sciences of the United States of America, cilt.107, sa.51, ss.22338-22343, 2010 (SCI-Expanded)
Stems of dicotyledonous plants consist of an outer epidermis, a cortex, a ring of secondarily thickened vascular bundles and interfascicular cells, and inner pith parenchyma cells with thin primary walls. It is unclear how the different cell layers attain and retain their identities. Here, we show that WRKY transcription factors are in part responsible for the parenchymatous nature of the pith cells in dicotyledonous plants. We isolated mutants of Medicago truncatula and Arabidopsis thaliana with secondary cell wall thickening in pith cells associated with ectopic deposition of lignin, xylan, and cellulose, leading to an ∼50% increase in biomass density in stemtissue of the Arabidopsismutants. Themutations are caused by disruption of stem-expressed WRKY transcription factor (TF) genes, which consequently up-regulate downstreamgenes encoding the NAM, ATAF1/2, and CUC2 (NAC) and CCCH type (C3H) zinc finger TFs that activate secondarywall synthesis. Direct binding of WRKY to the NAC gene promoter and repression of three downstream TFs were confirmed by in vitro assays and in planta transgenic experiments. Secondary wall-bearing cells form lignocellulosic biomass that is the source for second generation biofuel production. The discovery of negative regulators of secondary wall formationin pith opens up the possibility of significantly increasing the mass of fermentable cell wall components in bioenergy crops.