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Changes in the physiological characteristics of Panax ginseng embryogenic calli and molecular mechanism of ginsenoside biosynthesis under cold stress

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Abstract

Main conclusion

Short-term cold stress can induce the increased expression of key enzyme-encoding genes involved in secondary metabolite synthesis, thereby increasing secondary metabolite concentration.

Abstract

Cold stress is an ecologically limiting factor that strongly affects the physiological and biochemical properties of medicinal plants often resulting in changes of the secondary metabolic process. Ginsenosides are the main active ingredients in medicinal ginseng yet few studies exist on the effect of cold stress on the expression of ginsenosides or the molecular mechanism underlying its regulation. Here, we evaluated the effects of cold stress on the physiological characteristics and secondary metabolism of P. ginseng embryogenic calli. Physiological measurements and RNA-Seq analysis were used to dissect the metabolic and molecular responses of P. ginseng to cold conditions. We found that the dynamic accumulation of ginsenoside and various physiological indicators leads to homogenous adaptation to cold stress. Secondary metabolism of ginseng could be a compensation mechanism to facilitate its adaptation to cold stress. Combined with the changes in the endogenous hormone content, 9-cis-epoxycarotenoid dioxygenase (NCED), zeaxanthin epoxidase (ZEP), and short chain dehydrogenase (SDR) from the abscisic acid (ABA) synthesis pathway were identified as key mediators of this response. Thus, an appropriate degree of cold stress may promote accumulation of ginsenosides. Moreover, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR2), squalene epoxidase (SE1), squalene synthase (SS), dammarenediol synthase (DS-II), and β-alanine C-28 hydroxylase (CYP716A52v2) should be considered key mediators of the cold stress response and ginsenoside biosynthesis. During industrial production, short-term cold stress should be carried out on ginseng calli to improve the quality of its medicinal materials.

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Abbreviations

CAT:

Catalase

CYP716A52v2 :

β-Alanine C-28 hydroxylase

DS :

Dammarenediol synthase

HMGR :

3-Hydroxy-3-methylglutaryl-CoA reductase

JA:

Jasmonic acid

MDA:

Malondialdehyde

NCED:

9-Cis-epoxycarotenoid dioxygenase

POD:

Peroxidase

SA:

Salicylic acid

SDR:

Short chain dehydrogenase

SE :

Squalene epoxidase

SS1 :

Squalene synthase

SOD:

Superoxide dismutase

SP:

Soluble protein

SS:

Soluble sugars

ZEP:

Zeaxanthin epoxidase

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Acknowledgements

This work was supported by the major science and technology projects of the Jilin Province (20200504002YY), the National Key Research and Development Programs of China (2019YFC1710700), and the Chinese Agricultural Research System (grant number CARS-21).

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Authors and Affiliations

  1. Cultivation Base of State Key Laboratory for Ecological Restoration and Ecosystem Management of Jilin Province and Ministry of Science and Technology, College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun, 130118, Jilin, People’s Republic of China

    Tao Zhang, Yan Gao, Mei Han & Linmin Yang

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  1. Tao Zhang
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  2. Yan Gao
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  3. Mei Han
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  4. Linmin Yang
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Contributions

MH and LY conceived and designed the research. TZ performed the experiments and prepared the manuscript; YG completed the experimental data processing; all authors read and approved the manuscript.

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Correspondence to Mei Han or Linmin Yang.

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Communicated by Dorothea Bartels.

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Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PNG 66 KB) Fig. S1 GC content of the Saccharum officinarum unigenes.

Supplementary file2 (PNG 125 KB) Fig. S2 Analysis of T_4 vs C_4 differentially expressed gene GO enrichment

Supplementary file3 (PNG 126 KB) Fig. S3 Analysis of T_6 vs C_6 differentially expressed gene GO enrichment

Supplementary file4 (PNG 161 KB) Fig. S4 Analysis of T_4 vs C_4 differentially expressed gene KEGG enrichment

425_2020_3535_MOESM5_ESM.png

Supplementary file5 (PNG 175 KB) Fig. S5 Analysis of T_6 vs C_6 differentially expressed gene KEGG enrichment. Fig. S6 Validation of the transcriptome data using qRT-PCR. Vertical bars indicate the mean value ± SD from three independent experiments. *P < 0.05 and **P < 0.01 denote significant differences from the control, respectively.

425_2020_3535_MOESM6_ESM.xlsx

Supplementary file6 (XLSX 10 KB) Table S1 FPKM results of ten randomly selected P. ginseng calli unigenes identified during high-throughput transcriptome sequencing

Supplementary file7 (XLSX 10 KB) Table S2 Primers used in the qRT-PCR validation assays

Supplementary file8 (XLSX 18526 KB) Table S3 Differential gene expression

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Zhang, T., Gao, Y., Han, M. et al. Changes in the physiological characteristics of Panax ginseng embryogenic calli and molecular mechanism of ginsenoside biosynthesis under cold stress. Planta 253, 79 (2021). https://doi.org/10.1007/s00425-020-03535-7

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