Identification of key genes and active anti-inflammatory ingredients in Panax medicinal plants by climate-regulated callus culture combined with gene-component-efficacy gray correlation analysis

Abstract

Objective:We aimed to establish a novel strategy for identifying key genes and active anti-inflammatory ingredients in Panax medicinal plants.

Methods:First, fresh roots of 2-year-old Panax plants, including P. ginseng C. A. Mey., P. quinquefolium L., P. notoginseng (Burk.) F. H. Chen, P. japonicus C.A.Mey., P. japonicus Mey. var. major (Burk.) C. Y. Wu et K. M. Feng, were selected as explants, and callus formation was induced under three experimental temperatures (17, 24, and 30°C). Second, high-performance liquid chromatography-mass spectrometry was used to analyze the saponin content of the callus. Nitric oxide reduction efficacy was used for “component-efficacy” gray correlation analysis to find the active anti-inflammatory ingredients. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to determine the inflammatory factors and verify the active ingredients’ anti-inflammatory effects. Finally, qRT-PCR was used to detect the expression of key genes in the callus, and “gene-component” gray correlation analysis was used to examine the relationships between the regulatory pathway of the genes and the components.

Results:Among the three experimental temperatures (17, 24, and 30°C), the lowest temperature (17°C) is the most suitable for generating Panax callus. Lower-latitude native Panax notoginseng is more adaptable under high culture temperatures (24°C and 30°C) than other Panax plants. The ginsenoside contents of the callus of P. notoginseng and P. japonicus were the highest under similar climate conditions (17°C). Major anti-inflammatory components were G-Rh1, G-Rb1, G-Rg3, and G-Rh6/Floral-GKa. CYP76A47 contributed to the accumulation of anti-inflammatory components.

Conclusions:This study provides a strategy for the gene-component-efficacy correlational study of multi-component, multi-functional, and multi-purpose plants of the same genus.

Keywords

Anti-inflammatory ingredient / Gene-component-efficacy gray correlation analysis / HPLC-MS / Panax / qRT-PCR / Callus culture

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