Liu Qun,Li Li,Cheng Haitao,Yao Lixiang,Wu Jie,Huang Hui,Ning Wei,Kai Guoyin
Polyphenols are the main active components of the anti-inflammatory compounds in dandelion, and chlorogenic acid (CGA) is one of the primary polyphenols. However, the molecular mechanism underlying the transcriptional regulation of CGA biosynthesis remains unclear. Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase (HQT2) is the last rate-limiting enzyme in chlorogenic acid biosynthesis in Taraxacum antungense. Therefore, using the TaHQT2 gene promoter as a probe, a yeast one-hybrid library was performed, and a basic helix-loop-helix (bHLH) transcription factor, TabHLH1, was identified that shared substantial homology with Gynura bicolor DC bHLH1. The TabHLH1 transcript was highly induced by salt stress, and the TabHLH1 protein was localized in the nucleus. CGA and luteolin concentrations in TabHLH1-overexpression transgenic lines were significantly higher than those in the wild type, while CGA and luteolin concentrations in TabHLH1-RNA interference (RNAi) transgenic lines were significantly lower. Quantitative real-time polymerase chain reaction demonstrated that overexpression and RNAi of TabHLH1 in T. antungense significantly affected CGA and luteolin concentrations by upregulating or downregulating CGA and luteolin biosynthesis pathway genes, especially TaHQT2, 4-coumarate-CoA ligase (Ta4CL), chalcone isomerase (TaCHI), and flavonoid-3′-hydroxylase (TaF3′H). Dual-luciferase, yeast one-hybrid, and electrophoretic mobility shift assays indicated that TabHLH1 directly bound to the bHLH-binding motifs of proTaHQT2 and proTa4CL. This study suggests that TabHLH1 participates in the regulatory network of CGA and luteolin biosynthesis in T. antungense and might be useful for metabolic engineering to promote plant polyphenol biosynthesis.
WRKY transcription factors play a critical role in metabolism and stress responses in plants, but few WRKYs have been reported in Taraxacum antungense . Here, a multiple stress-inducible gene, TaWRKY14 , was isolated from T. antungense . TaWRKY14 was localized to the nuclei, and phylogenetic analysis indicated that TaWRKY14 shared the highest identity to WRKY40 in Salvia miltiorrhiza . TaWRKY14 was highly expressed in roots, and up-regulated by salt and drought stress, salicylic acid treatment and powdery mildew. TaWRKY14 -overexpressing transgenic lines had higher chlorogenic acid concentration and high expression of TaPAL1 . The yeast one-hybrid assay proved that TaWRKY14 bond to the W-box of proTaPAL1 . Additionally, field experiments showed that TaWRKY14 -overexpressing T. antungense lines have higher powdery mildew resistance than the wild-type. Our results demonstrated that TaWRKY14 can regulate CGA biosynthesis and play an important role in resistance to powdery mildew in T. antungense . TaWRKY14 transgenic T. antungense can be used for further evaluation as a new germplasm resource.