Abstract
Ginseng (Panax ginseng C.A. Mey.) is a precious Chinese traditional medicine, for which ginsenosides are the most important medicinal ingredients. Cytochrome P450 enzymes (CYP450) and their primary redox molecular companion NADPH cytochrome P450 reductase (CPR) play a key role in ginsenoside biosynthesis pathway. However, systematic studies of CPR genes in ginseng have not been reported. Numerous studies on ginsenoside synthesis biology still use Arabidopsis CPR (AtCPR1) as a reductase. In this study, we isolated two CPR genes (PgCPR1, PgCPR2) from ginseng adventitious roots. Phylogenetic tree analysis showed that both PgCPR1 and PgCPR2 are grouped in classⅡ of dicotyledonous CPR. Enzyme experiments showed that recombinant proteins PgCPR1, PgCPR2 and AtCPR1 can reduce cytochrome c and ferricyanide with NADPH as the electron donor, and PgCPR1 had the highest enzymatic activities. Quantitative real-time PCR analysis showed that PgCPR1 and PgCPR2 transcripts were detected in all examined tissues of Panax ginseng and both showed higher expression in stem and main root. Expression levels of the PgCPR1 and PgCPR2s were both induced after a methyl jasmonate (MeJA) treatment and its pattern matched with ginsenoside accumulation. The present investigation suggested PgCPR1 and PgCPR2 are associated with the biosynthesis of ginsenoside. This report will assist in future CPR family studies and ultimately improving ginsenoside production through transgenic engineering and synthetic biology.
Highlights
Blast results show that PgCPR1 and PgCPR2 are 99% similar to ginseng CPR1 (AHA50098.1) and ginseng CPR2 (AIC73829.1), which have been uploaded into the NCBI database
Conserved domains finder results show that both cytochrome P450 reductase (CPR) contain all functional domains that bind to Cytochrome P450 enzymes (CYP450) and interact with coenzyme NADPH, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD)
The results showed that the expression of PgCPR1 reached the maximum value in 100 μM methyl jasmonate (MeJA) treatments, and decreased significantly and
Summary
Ginsenoside is the most important medicinal active component in ginseng [3]. More and more research has found that ginsenoside can reduce oxidative stress response, protect the heart [4], slow down depression [5], prevent and treat Alzheimer’s disease [6,7,8], inhibit the growth of tumor cells [9,10,11,12] and other pharmacological effects. Ginseng, a crop with a long growth period and a continuous cropping obstacle are seriously limited by the growing environment in yield and quality. The contents of ginsenosides with unique pharmacological activities are very few [13]. Many researchers attempt to use functional gene identification and synthetic biology to improve the yield of ginsenoside
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