The plant-specific family of WUSCHEL (WUS)-related homeobox (WOX) transcription factors is key regulators of embryogenesis, meristem maintenance, and lateral organ development in flowering plants. The modern/WUS clade transcriptional repressor STENOFOLIA/LAMINA1(LAM1), and the intermediate/WOX9 clade transcriptional activator MtWOX9/NsWOX9 antagonistically regulate leaf blade expansion, but the molecular mechanism is unknown. Using transcriptome profiling and biochemical methods, we determined that NsCKX3 is the common target of LAM1 and NsWOX9 in Nicotiana sylvestris. LAM1 and NsWOX9 directly recognize and bind to the same cis-elements in the NsCKX3 promoter to repress and activate its expression, respectively, thus controlling the levels of active cytokinins in vivo. Disruption of NsCKX3 in the lam1 background yielded a phenotype similar to the knockdown of NsWOX9 in lam1, while overexpressing NsCKX3 resulted in narrower and shorter lam1 leaf blades reminiscent of NsWOX9 overexpression in the lam1 mutant. Moreover, we established that LAM1 physically interacts with NsWOX9, and this interaction is required to regulate NsCKX3 transcription. Taken together, our results indicate that repressor and activator WOX members oppositely regulate a common downstream target to function in leaf blade outgrowth, offering a novel insight into the role of local cytokinins in balancing cell proliferation and differentiation during lateral organ development.
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Zhang, Fei ; Wang, Hui ; Kalve, Shweta ; Wolabu, Tezera W. ; Nakashima, Jin ; Golz, John F. ; Tadege, Million ( , New Phytologist)
Summary Plant lateral organ development is a complex process involving both transcriptional activation and repression mechanisms. The
WOX transcriptional repressorWOX 1/STF , theLEUNIG (LUG ) transcriptional corepressor and theANGUSTIFOLIA 3 (AN 3) transcriptional coactivator play important roles in leaf blade outgrowth and flower development, but how these factors coordinate their activities remains unclear. Here we report physical and genetic interactions among these key regulators of leaf and flower development.We developed a novel
in planta transcriptional activation/repression assay and suggest thatLUG could function as a transcriptional coactivator during leaf blade development.Mt
LUG physically interacts with MtAN 3, and this interaction appears to be required for leaf and flower development. A single amino acid substitution at position 61 in theSNH domain of MtAN 3 protein abolishes its interaction with MtLUG , and its transactivation activity and biological function. Mutations inlug andan3 enhanced each other's mutant phenotypes. Both thelug and thean3 mutations enhanced thewox1 prs leaf and flower phenotypes inArabidopsis .Our findings together suggest that transcriptional repression and activation mediated by the
WOX ,LUG andAN 3 regulators function in concert to promote leaf and flower development, providing novel mechanistic insights into the complex regulation of plant lateral organ development.