科研之友
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摘要
Objective Based on the Panax notoginseng root transcriptome database, the key enzyme cytochrome P450 (CYP450) gene of P. notoginseng saponin synthesis was cloned and bioinformatically analyzed to clarify its response to the induction of arbuscular mycorrhizal fungi (AMF) and to lay a foundation for the further study on catalytic properties. Methods The clone of PnCyp450_3 ORF and the construction of cloning vector were carried out according to the corresponding kit instruction. The positive clone was gained by chemical conversion and ampicillin screening. Bioinformatics analysis and phylogenetic tree were generated by a series of online tools or software, while gene expression characteristics were analyzed by real-time fluorescence quantitative (qPCR). Results A P. notoginseng cytochrome P450 superfamily gene, PnCyp450_3, was successfully cloned with the open reading frame (ORF) of 1617 bp, encoding 538 amino acids. Predicted molecular weight and isoelectric point (pI) was 59 690 and 8.50, respectively. The results of tissue expression characteristics showed that the expression level of PnCyp450_3 was significantly up-regulated by Glomus intraradices (P < 0.05) in the rhizomes and fibrous roots with relative expression levels of 1.85 and 1.22. Conclusion PnCyp450_3, a CYP450 superfamily gene encoding 538 amino acids, was significantly up-regulated in both primary and fibrous roots in response to the induction of G. intraradices. This study provides data to support an in-depth study of the regulatory mechanism of biological factors to key enzyme-encoding genes on the notoginsenoside synthesis, which has important reference value.
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