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Molecular cloning and transgenic characterization of the genes encoding chalcone synthase and chalcone isomerase from the Tibetan herbal plant Mirabilis himalaica
Author(s) -
Lan Xiaozhong,
Quan Hong,
Xia Xinli,
Yin Weilun,
Zheng Weilie
Publication year - 2015
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1376
Subject(s) - chalcone synthase , biology , chalcone isomerase , gene , transgene , genetically modified crops , chalcone , cloning (programming) , molecular cloning , isomerase , biosynthesis , microbiology and biotechnology , biochemistry , botany , gene expression , chemistry , stereochemistry , computer science , programming language
Mirabilis himalaica is an endangered medicinal plant species in the Tibetan Plateau. The two genes respectively encoding chalcone synthase ( MhCHS ) and chalcone isomerase ( MhCHI ) were isolated and characterized from M. himalaica . The sequence analysis revealed that the two genes were similar with their corresponding homologous genes in other plants. The tissue profiles showed that both MhCHS and MhCHI had higher expression levels in roots than in stems and leaves. Transgenic hairy root cultures respectively with overexpressing MhCHS and MhCHI were established. The genomic PCR detection confirmed the authority of transgenic hairy root lines, in which either MhCHS or MhCHI expression levels were much higher than that in non‐transgenic hairy root line. Finally, the HPLC detection results demonstrated that the rotenoid contents in MhCHS / MhCHI ‐transformed hairy root lines were enhanced. This study provided two candidate genes that could be used to genetic engineering rotenoid biosynthesis in M. himalaica and an alternative method to produce rotenoid using transgenic hairy root cultures.