PJB-2018-531
Enhancing glycyrrhizic acid accumulation by over-expressing β-amyrin synthase gene (GuBAS) root-specifically in hairy roots of Glycyrrhiza uralensis
Yan-chao Yin
Abstract
Glycyrrhizia uralensis is one of the most widely-used herbal medicines in China. Glycyrrhizic acid (GA) is its important marker component, which is biosynthesized by many enzymes. Among them β-amyrin synthase is responsible for triterpene skeleton formation. In this paper, we tried to analyze the influence of β-amyrin synthase gene (GuBAS) on the accumulation of GA. Tobacco root-specific promoter TobRB7 and GuBAS cDNA were obtained and combined with linearized pCAMBIA1305.1 to construct root-specific plant expression vector which was later transformed into Agrobacterium rhizogenes ACCC10060 by electrotransformation. The cotyledons and hypocotyls of G. uralensis were infected by the recombinant A. rhizogenes ACCC10060 to induce hairy roots. The PCR and sequencing results both show that 3 transgenic hairy root lines were got. The copy number of GuBAS in these transgenic hairy roots was intended by qRT-PCR to be 3, 7, and 4. The GA content was assayed by HPLC, and the results showed that GA was present in the three transgenic hairy roots, but absent in wild hairy roots. Therefore, over-expressing GuBAS root-specifically in hairy roots of G. uralensis enhanced GA accumulation. This paper will contribute to the evaluation of GuBAS functions, GA production, and licorice molecular breeding as well.
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