In vitro evaluation of hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase expression and regulation in Taraxacum antungense in relation to 5-caffeoylquinic acid production. (June 2019)

• Record Type: Journal Article
• Authors: Liu, Qun; Yao, Lixiang; Xu, Yachen; Cheng, Haitao; Wang, Weiting; Liu, Zijia; Liu, Jia; Cui, Xin; Zhou, Yujie; Ning, Wei
• Is Part Of: Phytochemistry. Volume 162(2019)
• Page Start: 148
• Page End: 156
• Publication Date: 2019-06
• Subjects: Taraxacum antungense Kitag. -- Compositae -- Tissue inoculation -- 5-caffeoylquinic acid -- TaHQT; CGA Chlorogenic acids -- FPNI-PCR Fusion primer and nested integrated PCR -- HQT Hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase -- HPLC High-performance liquid chromatography -- Kan Kanamycin -- MS Murashige and Skoog -- qRT-PCR Quantitative reverse transcription PCR -- RNAi RNA interference -- SDS-PAGE Sodium dodecyl sulphate-polyacrylamide gel electrophoresis -- PBS Potassium phosphate buffer -- dry wt dry weight -- fr. wt Fresh weight; Botanical chemistry -- Periodicals; Biochemistry -- Periodicals; Botany -- Periodicals; Chimie végétale -- Périodiques; 572.2
• DOI: 10.1016/j.phytochem.2019.02.014
• Languages: English
• ISSNs: 0031-9422

Abstract

Abstract: Chlorogenic acids (CGA; including 5-caffeoylquinic acid and its regio-isomers) in Taraxacum antungense Kitag. have antioxidant and anti-inflammatory properties and exert other pharmacological effects. T. antungense hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (TaHQT)1 and TaHQT2, which belong to the BAHD acyltransferase family, are candidates for synthesizing 5-caffeoylquinic acid and that have not been extensively characterized. In this study, we cloned the TaHQT1 and TaHQT2 genes and analysed the properties of the expressed enzymes both in vitro and in vivo . Quantitative reverse transcription PCR analysis revealed that TaHQT1 was highly expressed in the root, whereas the strongest TaHQT2 expression was observed in T. antungense leaves. In Nicotiana benthamiana leaf cells, TaHQT1 and TaHQT2 were localized at the cell periphery as well as in the cytoplasm and nucleus. The 5-caffeoylquinic acid concentrations in T. antungense calli were reduced by TaHQT1 and TaHQT2 knockdown relative to the control. Conversely, inoculation of T. antungense plants tissues with recombinant TaHQT1 and TaHQT2 increased 5-caffeoylquinic acid levels in situ . These in vitro and in vivo findings demonstrate that both HQTs are involved in regulating 5-caffeoylquinic acid biosynthesis in T. antungense, which can be exploited to increase 5-caffeoylquinic acid production in plants for medicinal or other beneficial purposes. Graphical abstract: Picture of the flowering Taraxacum antungense Kitag., (Dandong dandelion) (A); the basic biosynthesis pathway of 5-caffeoylquinic acid in plants (B) requiring HQTs.Image 1 Highlights: TaHQT1 and TaHQT2 RNAi calli of Taraxacum antungense were generated and evaluated. TaHQT1 was localized at the cell periphery of N. benthamiana leaf cells. TaHQT2 was localized in the cytoplasm and nucleus of N. benthamiana leaf cells. Plant tissue treatment with recombinant HQT reveals its effect on 5-caffeoylquinic acid biosynthesis.