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

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. Graphical abstract: http://links.lww.com/AHM/A38

[1]  G. Pan,et al.  New Insights Into Tissue Culture Plant-Regeneration Mechanisms , 2022, Frontiers in Plant Science.

[2]  Yen On Chan,et al.  Candidate Genes Modulating Reproductive Timing in Elite US Soybean Lines Identified in Soybean Alleles of Arabidopsis Flowering Orthologs With Divergent Latitude Distribution , 2022, Frontiers in Plant Science.

[3]  Zhanghua Wu,et al.  Design and synthesis of adamantyl-substituted flavonoid derivatives as anti-inflammatory Nur77 modulators: Compound B7 targets Nur77 and improves LPS-induced inflammation in vitro and in vivo. , 2022, Bioorganic chemistry.

[4]  Yonghong Guan,et al.  Indole derivative XCR-5a alleviates LPS-induced inflammation in vitro and in vivo , 2021, Immunopharmacology and immunotoxicology.

[5]  Sai Guna Ranjan Gurazada,et al.  The evolutionary history of small RNAs in Solanaceae , 2021, bioRxiv.

[6]  Qinghe Zhang,et al.  Changes in the Leaf Physiological Characteristics and Tissue-Specific Distribution of Ginsenosides in Panax ginseng During Flowering Stage Under Cold Stress , 2021, Frontiers in Bioengineering and Biotechnology.

[7]  Shujuan Zhao,et al.  Ginsenosides in Panax genus and their biosynthesis , 2021, Acta pharmaceutica Sinica. B.

[8]  Zhihua Zhou,et al.  The unprecedented diversity of UGT94-family UDP-glycosyltransferases in Panax plants and their contribution to ginsenoside biosynthesis , 2020, Scientific Reports.

[9]  Lan Zhang,et al.  Ginsenoside Rg1 prevent and treat inflammatory diseases: A review. , 2020, International immunopharmacology.

[10]  Jing Wang,et al.  Anticoagulant active ingredients identification of total saponin extraction of different panax medicinal plants based on grey relational analysis combined with UPLC-MS and molecular docking. , 2020, Journal of ethnopharmacology.

[11]  D. Im.  Pro-Resolving Effect of Ginsenosides as an Anti-Inflammatory Mechanism of Panax ginseng , 2020, Biomolecules.

[12]  M. Pathak.  Study on secondary metabolites produced from callus cultures of Nicotiana tabacum by plant tissue culture techniques , 2019, Journal of Biotechnology.

[13]  A. Wong,et al.  Chemical Structures and Pharmacological Profiles of Ginseng Saponins , 2019, Molecules.

[14]  Ze-Min Yang,et al.  Verification of miRNAs in ginseng decoction by high-throughput sequencing and quantitative real-time PCR , 2019, Heliyon.

[15]  Zibo Li,et al.  Transcriptome analysis of Sclerotinia ginseng and comparative analysis with the genome of Sclerotinia sclerotiorum , 2019, Physiological and Molecular Plant Pathology.

[16]  T. Efferth.  Biotechnology Applications of Plant Callus Cultures , 2019, Engineering.

[17]  Byoung Ryong Jeong,et al.  In vitro cultivation of Panax ginseng C.A. Meyer , 2018, Industrial Crops and Products.

[18]  Hongling Zhang,et al.  Longitudinal expression patterns of HMGR, FPS, SS, SE and DS and their correlations with saponin contents in green-purple transitional aerial stems of Panax notoginseng , 2018, Industrial Crops and Products.

[19]  Shi-hui Wang,et al.  Characterization of UDP-Glycosyltransferase Involved in Biosynthesis of Ginsenosides Rg1 and Rb1 and Identification of Critical Conserved Amino Acid Residues for Its Function. , 2018, Journal of agricultural and food chemistry.

[20]  Daniel S. Park,et al.  Genome and evolution of the shade‐requiring medicinal herb Panax ginseng , 2018, Plant biotechnology journal.

[21]  P. Zhu,et al.  Progress on the Studies of the Key Enzymes of Ginsenoside Biosynthesis , 2018, Molecules.

[22]  Chang-bao Chen,et al.  Multicomponent assessment and ginsenoside conversions of Panax quinquefolium L. roots before and after steaming by HPLC-MSn , 2017, Journal of ginseng research.

[23]  Chaoyin Chen,et al.  Enhancement of triterpenoid saponins biosynthesis in Panax notoginseng cells by co-overexpressions of 3-hydroxy-3-methylglutaryl CoA reductase and squalene synthase genes , 2017 .

[24]  En-Hua Xia,et al.  The Medicinal Herb Panax notoginseng Genome Provides Insights into Ginsenoside Biosynthesis and Genome Evolution. , 2017, Molecular plant.

[25]  Hideyuki Suzuki,et al.  CYP716A179 functions as a triterpene C-28 oxidase in tissue-cultured stolons of Glycyrrhiza uralensis , 2017, Plant Cell Reports.

[26]  R. Whetten,et al.  Ecological genomics of local adaptation in Cornus florida L. by genotyping by sequencing , 2016, Ecology and evolution.

[27]  R. Saunders,et al.  Reproductive resource partitioning in two sympatric Goniothalamus species (Annonaceae) from Borneo: floral biology, pollinator trapping and plant breeding system , 2016, Scientific Reports.

[28]  D. Guo,et al.  Identification and differentiation of Panax ginseng, Panax quinquefolium, and Panax notoginseng by monitoring multiple diagnostic chemical markers , 2016, Acta pharmaceutica Sinica. B.

[29]  Ying-ping Wang,et al.  Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC-DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS , 2015, Journal of ginseng research.

[30]  Deok-Chun Yang,et al.  Biosynthesis and biotechnological production of ginsenosides. , 2015, Biotechnology advances.

[31]  Xiaoyan Chen,et al.  Determination of ginsenoside compound K in human plasma by liquid chromatography–tandem mass spectrometry of lithium adducts , 2015, Acta pharmaceutica Sinica. B.

[32]  Zhihua Zhou,et al.  Production of bioactive ginsenosides Rh2 and Rg3 by metabolically engineered yeasts. , 2015, Metabolic engineering.

[33]  J. Thevelein,et al.  Unraveling the triterpenoid saponin biosynthesis of the African shrub Maesa lanceolata. , 2014, Molecular plant.

[34]  Wan-ying Wu,et al.  Saponins in the genus Panax L. (Araliaceae): a systematic review of their chemical diversity. , 2014, Phytochemistry.

[35]  S. Ko,et al.  Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by γ-irradiation (60Co) of adventitious roots , 2014, Journal of ginseng research.

[36]  M. Rogero,et al.  A High-Fat Diet Increases IL-1, IL-6, and TNF-α Production by Increasing NF-κB and Attenuating PPAR-γ Expression in Bone Marrow Mesenchymal Stem Cells , 2013, Inflammation.

[37]  Zai-Qun Liu.  Chemical insights into ginseng as a resource for natural antioxidants. , 2012, Chemical reviews.

[38]  Kazuki Saito,et al.  CYP716A subfamily members are multifunctional oxidases in triterpenoid biosynthesis. , 2011, Plant & cell physiology.

[39]  N. Graham,et al.  Medicago truncatula CYP716A12 Is a Multifunctional Oxidase Involved in the Biosynthesis of Hemolytic Saponins[W] , 2011, Plant Cell.

[40]  K. Leung,et al.  Pharmacology of ginsenosides: a literature review , 2010, Chinese medicine.

[41]  Hong Xu,et al.  Differentiation of the root of Cultivated Ginseng, Mountain Cultivated Ginseng and Mountain Wild Ginseng using FT-IR and two-dimensional correlation IR spectroscopy , 2008 .

[42]  Susan C. Roberts,et al.  Pharmaceutically active natural product synthesis and supply via plant cell culture technology. , 2008, Molecular pharmaceutics.

[43]  S. Akira,et al.  Toll-like receptors and innate immunity , 2006, Journal of Molecular Medicine.

[44]  Y. Ebizuka,et al.  Mutational Studies on Triterpene Synthases: Engineering Lupeol Synthase into β-Amyrin Synthase , 2000 .

[45]  T. Thorpe.  History of plant tissue culture , 2007, Molecular biotechnology.

[46]  W. Soh,et al.  Origin of somatic embryo induced from cotyledons of zygotic embryos at various developmental stages of ginseng , 1994 .