Overexpression of G10H and ORCA3 in the hairy roots of Catharanthus roseus improves catharanthine production.
A number of genes that function in the terpenoid indole alkaloids (TIAs) biosynthesis pathway have been identified in Catharanthus roseus. Except for the geraniol 10-hydroxylase (G10H) gene, which encodes a cytochrome P450 monooxygenase, several of these genes are up-regulated by ORCA3, a jasmonate-responsive APETALA2 (AP2)-domain transcript factor. In this study, the G10H gene was transformed independently, or co-transformed with ORCA3 into C. roseus, using Agrobacterium rhizogenes MSU440. Hairy root clones expressing the G10H gene alone, or both the G10H and ORCA3 genes, were obtained. Alkaloid accumulation level analyses showed that all transgenic clones accumulated more catharanthine, with the highest accumulation level in the transgenic clone OG12 (6.5-fold higher than that of the non-expression clone). Following treatment with ABA, accumulation of catharanthine reached 1.96 mg/g DW in the transgenic clone OG12. The expression levels of TIAs biosynthesis genes in transgenic and non-transgenic clones were also investigated.
National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China.
Cui-Ting
CT
Wang
Hua
H
Liu
Xiao-Shu
XS
Gao
Hong-Xia
HX
Zhang
D000040:Abscisic Acid / Q000494:pharmacology; D030008:Catharanthus / Q000201:enzymology / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014748:Vinca Alkaloids / Q000096:biosynthesis*
D018744:DNA, Plant; C410983:ORCA3 protein, plant; D010940:Plant Proteins; D014157:Transcription Factors; D014748:Vinca Alkaloids; D000040:Abscisic Acid; D003577:Cytochrome P-450 Enzyme System; C081675:geraniol 10-hydroxylase; C017836:catharanthine