Chlormequat chloride-responsive VviDOF3.4-VviLBD36 module regulates adventitious root formation in grapevine

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The main mode of reproduction in grapevine is asexual reproduction, and the adventitious root (AR) formation is essential for asexual reproduction in grapevine. Chlormequat chloride (CCC) is a gibberellin antagonist that can regulate plant growth and development. However, CCC has rarely been studied for its effect on AR formation. This study found that CCC treatment significantly promoted the formation of AR in 'Thompson Seedless' (Vitis vinifera L. cv. Thompson Seedless). Transcriptome sequencing and in-depth analysis of AR from CCC-treated plants revealed that VviLBD36 is a key factor involved in AR formation. VviLBD36 was significantly upregulated after CCC treatment, and its overexpression markedly increased the number of AR and lateral root (LR) in Arabidopsis. Yeast one-hybrid and dual-luciferase assays demonstrated that the VviDOF3.4 directly binds to the promoter of VviLBD36 and activates its expression. Additionally, overexpression of VviDOF3.4 also promoted LR development in Arabidopsis. In summary, after CCC treatment, VviDOF3.4 promotes AR formation in grapevine by activating the transcription of VviLBD36. This study elucidates the role of the VviDOF3.4-VviLBD36 module in regulating grapevine AR formation in response to CCC, providing a theoretical basis for in-depth analysis of the molecular mechanisms by which CCC affects AR formation. Key message VviLBD36 regulates adventitious root formation in grapevine.