Transcriptome analysis of Panax zingiberensis identifies genes encoding oleanolic acid glucuronosyltransferase involved in the biosynthesis of oleanane-type ginsenosides
Oleanolic acid glucuronosyltransferase (OAGT) genes synthesizing the direct precursor of oleanane-type ginsenosides were discovered. The four recombinant proteins of OAGT were able to transfer glucuronic acid at C-3 of oleanolic acid that yields oleanolic acid 3-O-β-glucuronide.
Ginsenosides are the primary active components in the genus Panax, and great efforts have been made to elucidate the mechanisms underlying dammarane-type ginsenoside biosynthesis. However, there is limited information on oleanane-type ginsenosides. Here, high-performance liquid chromatography analysis demonstrated that oleanane-type ginsenosides (particularly ginsenoside Ro and chikusetsusaponin IV and IVa) are the abundant ginsenosides in Panax zingiberensis, an extremely endangered Panax species in southwest China. These ginsenosides are derived from oleanolic acid 3-O-β-glucuronide, which may be formed from oleanolic acid catalyzed by an unknown oleanolic acid glucuronosyltransferase (OAGT). Transcriptomic analysis of leaves, stems, main roots, and fibrous roots of P. zingiberensis was performed, and a total of 46,098 unigenes were obtained, including all the identified homologous genes involved in ginsenoside biosynthesis. The most upstream genes were highly expressed in the leaves, and the UDP-glucosyltransferase genes were highly expressed in the roots. This finding indicated that the precursors of ginsenosides are mainly synthesized in the leaves and transported to different parts for the formation of particular ginsenosides. For the first time, enzyme activity assay characterized four genes (three from P. zingiberensis and one from P. japonicus var. major, another Panax species with oleanane-type ginsenosides) encoding OAGT, which particularly transfer glucuronic acid at C-3 of oleanolic acid to form oleanolic acid 3-O-β-glucuronide. Taken together, our study provides valuable genetic information for P. zingiberensis and the genes responsible for synthesizing the direct precursor of oleanane-type ginsenosides.
KeywordsBiosynthesis Ginsenosides Oleanolic acid glucuronosyltransferase (OAGT) Transcriptome Triterpenoid saponins
Oleanolic acid glucuronosyltransferase
Reads per kilobase per million mapped reads
Uridine diphosphate-dependent glycosyltransferases
Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry
This work was supported by the National Natural Science Foundation of China (Grant no. U1402262), National Key R & D Plan (no. 2017YFC1702500), the Major Science and Technique Programs in Yunnan Province (Grant no. 2016ZF001), the Natural Science Foundation of Yunnan Province (Grant no. 2015FB147) and Guangxi Scientific Research and Technology Development Program (Guikezhong 14124002-1). This paper was dedicated to Mrs. Zhang, Professor Zhang’s wife, who suffered ataxia which caused serious lateral curvature. She could not stand, walk and sleep for many years. If you can help her, please contact firstname.lastname@example.org.
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