title: [Enhancement of tropane alkaloids production in transgenic hair roots of Atropa belladonna by overexpressing endogenous genes AbPMT and AbH6H].
abstract: Atropa belladonna L. is the officially medicinal plant species and the main commercial source of scopolamine and hyoscyamine in China. In this study, we reported the simultaneous overexpression of two functional genes involved in biosynthesis of scopolamine, which respectively encoded the upstream key enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53) and the downstream key enzyme hyoscyamine 6beta-hydroxylase (H6H; EC 1.14.11.11) in transgenic hair root cultures of Atropa belladonna L. HPLC results suggested that four transgenic hair root lines produced higher content of scopolamine at different levels compared with nontransgenic hair root cultures. And scopolamine content increased to 8.2 fold in transgenic line PH2 compared with that of control line; and the other four transgenic lines showed an increase of scopolamine compared with the control. Two of the transgenic hair root lines produced higher levels of tropane alkaloids, and the content increased to 2.7 fold in transgenic line PH2 compared with the control. The gene expression profile indicated that both PMT and H6H expressed at a different levels in different transgenic hair root lines, which would be helpful for biosynthesis of scopolamine. Our studies suggested that overexpression of A. belladonna endogenous genes PMT and H6H could enhance tropane alkaloid biosynthesis.
authors.affiliation: Chongqing Engineering and Technology Center for Sweetpotato, School of Life Sciences, Southwest University, Chongqing 400715, China.
authors.forename: Shi-Ping
authors.initials: SP
authors.lastname: Long
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.forename: Ya-Xiong
authors.initials: YX
authors.lastname: Wang
authors.forename: Chun-Xian
authors.initials: CX
authors.lastname: Yang
authors.forename: Xiao-Zhong
authors.initials: XZ
authors.lastname: Lan
authors.forename: Zhi-Hua
authors.initials: ZH
authors.lastname: Liao
mesh_terms: D001532:Atropa belladonna / Q000201:enzymology / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D064692:Hyoscyamine / Q000378:metabolism; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D010946:Plants, Medicinal / Q000201:enzymology / Q000235:genetics; D012601:Scopolamine / Q000378:metabolism*; D058615:Synthetic Biology; D014326:Tropanes / Q000378:metabolism*
chemical_list: D014326:Tropanes; D012601:Scopolamine; D006899:Mixed Function Oxygenases; C052273:hyoscyamine (6S)-dioxygenase; D008780:Methyltransferases; C112598:putrescine N-methyltransferase; D064692:Hyoscyamine