Binding proteins PnCOX11 and PnDCD strongly respond to GA and ABA in Panax notoginseng

Panax notoginseng saponins (PNS) are one of the main active ingredients of Panax notoginseng, a representative plant of the genus Panax. However, the detailed regulation mechanism of PNS biosynthesis remains elusive. Therefore, a sequence of upstream promoters of PnSS and PnSE were cloned and analyzed firstly. GUS quantitative results showed that the upstream promoters could specifically and significantly respond to exogenous GA and ABA signals. To further identify the binding proteins that respond to peripheral hormones, PnCOX11 and PnDCD were screened and identified from the P. notoginseng cDNA library. The Y1H experiment verified the interaction between the above two binding proteins and the promoters. Several online software was used to analyze the domains, secondary structures, three-dimensional structures, and phylogenetic trees of the two binding proteins. Subcellular localization analysis exhibited that PnCOX11 was mainly located in the chloroplast, while PnDCD was located in the cytoplasm and nucleus. Prokaryotic expression demonstrated that the recombinant proteins had a high concentration under the induction of IPTG. This study can provide a fundamental date for the subsequent thorough investigation of the transcription regulatory mechanism of PNS biosynthesis.