BolANT3 Positively Regulates Indolic Glucosinolate Accumulation by Transcriptionally Activating BolCYP83B1 in Cabbage

Indolic glucosinolates are a group of plant secondary metabolites found in Brassica vegetables, and their breakdown products could act as important anti-cancer and defense compounds against biotic stresses. Transcriptional regulation plays a key role in modulating the biosynthesis of indolic glucosinolates in the model plant Arabidopsis, but little is known about the transcriptional regulatory landscape of these glucosinolates in Brassica vegetables. In this study, we selected and functionally validated the important biosynthetic gene BolCYP83B1 from the indolic glucosinolate pathway in cabbage. Through a yeast one-hybrid assay, we systemically screened and identified upstream regulators of BolCYP83B1 in cabbage with BolANTs as the top candidates for further functional validation. Two homologs of BolANTs, BolANT1 and BolANT3, were confirmed to bind the promoter of BolCYP83B1 via both a yeast one-hybrid assay and an LUC assay. The overexpression of BolANT3 in cabbage significantly increased the accumulation of indolic glucosinolates, while the virus-induced gene silencing (VIGS) of BolANT3 significantly reduced the accumulation of indolic glucosinolates in cabbage. Our work provides valuable insights into the transcriptional regulatory mechanisms of indolic glucosinolates in Brassica vegetables.