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Overexpression of geraniol 10-hydroxylase from Panax ginseng conferred enhanced resistance to Pseudomonas syringae in Arabidopsis

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Abstract

Geraniol, the precursor of terpenoid indole alkaloids can be converted to the 10-hydroxy geraniol by the function of geraniol 10-hydroxylase. In our study, for the first time, a full-length cytochrome P450 monooxygenase (P450) geraniol 10-hydroxylase (PgCYP76C9) cDNA was isolated and characterized from Panax ginseng Meyer. The gene has an open reading frame (ORF) of 1503 base pairs and encodes a precursor protein of 501 amino acids residues. The calculated molecular mass of the protein is approximately 56.3 kDa with a predicated isoelectric point of 8.45. Amino acid identities between PgG10H and other P450s of the CYP76 family in the database had revealed that the deduced amino acid of PgG10H sharing a higher sequence homology with geraniol 10-hydroxylase-like proteins encoded by Cinchona calisaya and Lonicera japonica. We implemented a molecular modeling method to evaluate the possible interaction of geraniol with PgG10H active site. Our finding showed that the geraniol was the potential ligand for PgG10H in P. ginseng. Expression of PgG10H gene was tissue-regulated and showed high expression in 3-year-old ginseng flowers and roots. Expression of PgG10H was differentially induced in ginseng, not only during Pseudomonas syringae infection and wounding but also after exposure to methyl jasmonate and salt stress. Furthermore, overexpression of the newly identified ginseng geraniol 10-hydroxylase P450 gene in Arabidopsis caused terpenoid indole alkaloid dihydrositsirikine production and also conferred enhanced resistance to P. syringae.

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Abbreviations

cDNA:

Complementary DNA

CFP:

Cyan fluorescent protein

DMAPP:

Dimethylallyl diphosphate

G10H:

Geraniol 10-hydroxylase

GC-MS:

Gas chromatography mass spectrometry

GPP:

Geranyl diphosphate

GPS:

Geranyl diphosphate synthase

IPP:

Isopentenyl diphosphate

MJ:

Methyl jasmonate

MEP:

Methylerythritol phosphate

MVA:

Mevalonate

P450:

Cytochrome P450

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

TIA:

Terpenoid indole alkaloid

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Acknowledgments

This research was supported by iPET (312064-03-1-HD040), Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Author contributions

S.R.D Balusamy designed the experiments and performed most of the experiments. S. Rahimi provided technical assistance and wrote the paper. K. Senthil implemented molecular modeling, and D.C. Yang supervised and Y.G. Cho contributed to the writing.

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    Authors and Affiliations

    1. Department of Oriental Medicinal Biotechnology, College of Life Sciences, Kyung Hee University, Yongin, 446-701, Republic of Korea

      Sri Renuka Devi Balusamy & Deok-Chun Yang

    2. Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea

      Sri Renuka Devi Balusamy

    3. Graduate School of Biotechnology and Ginseng Bank, College of Life Sciences, Kyung Hee University, Yongin, 446-701, Republic of Korea

      Shadi Rahimi & Deok-Chun Yang

    4. Department of Crop Science, Chungbuk National University, Cheongju, 28644, Republic of Korea

      Shadi Rahimi & Yong-Gu Cho

    5. Avinashilingam Deemed University for Women, Coimbatore, 641 043, India

      Kalaiselvi Senthil

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    1. Sri Renuka Devi Balusamy
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    Correspondence to Deok-Chun Yang.

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    Sri Renuka Devi Balusamy and Shadi Rahimi have contributed equally to this work.

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    Balusamy, S.R.D., Rahimi, S., Cho, YG. et al. Overexpression of geraniol 10-hydroxylase from Panax ginseng conferred enhanced resistance to Pseudomonas syringae in Arabidopsis. Plant Growth Regul 81, 305–316 (2017). https://doi.org/10.1007/s10725-016-0207-6

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