Skip to main content
SpringerLink
Log in

Differential DNA methylation in regulation of deacetylvindoline-4-O-acetyl transferase (DAT) gene in Catharanthus roseus

  • Original Article
  • Published:
Journal of Plant Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

The present study investigates the role of DNA methylation in regulation of deacetylvindoline-4-O-acetyltransferase (DAT) gene in Madagascar periwinkle (Catharanthus roseus). Expression analysis showed significant upregulated DAT transcript (20 fold), demeter like (1.37 fold) and repressor of silencing-1 (1.47 fold) in leaf tissues of C. roseus as compared to root tissues. Methylases such as domain rearranged methyltransferase and chromomethyl transferase were downregulated suggesting involvement of DNA methylation in DAT regulation. DNA methylation signatures on DAT gene were further evaluated after treatment of leaves from C.roseus with 5-azacytidine (20 μM) for 5 and 10 days. Promoter DNA (− 11 to − 378 region) methylation analysis showed hypermethylation on two CHG and three CHH sites after day 10 of treatment. Qualitative methylation specific PCR of the coding region (+ 153 to + 322 region) showed hypomethylation after 10 days of 5-azacytidine treatment. Expression analysis showed upregulated DAT gene expression in the leaves 10 days post treatment, which correlated with changes observed in DNA methylation. Moreover, genomic 5-methylcytosine levels decreased from 0.65 to 0.33% after 10 days of 5-azacytidine treatment. However, no significant change was observed after 5 day of 5-azacytidine treatment as compared to control across different observational end points. The study clearly suggests that DAT gene expression may be regulated by differential DNA methylation in the promoter and coding regions. Further, hypomethylating agent such as 5-azacytidine holds potential to regulate the expression of DAT that regulates the terminal step in the indole alkaloid pathway leading to vindoline synthesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Source of Sequence, Wang et al. (2010)

Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

CMT3:

Chromomethyl transferase 3

DAT:

Deacetylvindoline-4-O-acetyl transferase

DML1:

Demeter like 1

DRM2:

Domain rearranged methyltransferase

MeJA:

Methyl jasmonate

ROS1:

Repressor of silencing 1

STR:

Strictosidine synthase

STS:

Stilbene synthase

References

  • Arfmann HA, Kohl W, Wray V (1985) Effect of 5-azacytidine on the formation of secondary metabolites in Catharanthus roseus cell suspension cultures. J Biosci 40:21–25

    Google Scholar 

  • Bahar Halpern K, Vana T, Walker MD (2014) Paradoxical role of DNA methylation in activation of FoxA2 gene expression during endoderm development. J Biol Chem 289(34):23882–23892

    Article  Google Scholar 

  • Bock C, Reither S, Mikeska T, Paulsen M, Walter J, Lengauer T (2005) BiQ analyzer: visualization and quality control for DNA methylation data from bisulfate sequencing. Bioinformatics 21(21):4067–4068

    Article  CAS  Google Scholar 

  • Furner IJ, Matzke M (2011) Methylation and demethylation of the Arabidopsis genome. Curr Opin Plant Biol 14(2):137–141

    Article  CAS  Google Scholar 

  • Giri AK, Aittokallio T (2019) DNMT inhibitors increase methylation in the cancer genome. Front Pharmacol 10:385

    Article  CAS  Google Scholar 

  • Hagemann S, Heil O, Lyko F, Brueckner B (2011) Azacytidine and decitabine induce gene-specific and non-random DNA demethylation in human cancer cell lines. PLoS ONE 6(3):e17388

    Article  CAS  Google Scholar 

  • Hauser MT, Aufsatz W, Jonak C, Luschnig C (2011) Transgenerational epigenetic inheritance in plants. Biochim Biophys Acta Gene Regul Mechan 1809:459–468

    Article  CAS  Google Scholar 

  • Khanuja SP, Shasany AK, Darokar M, Kumar S (1999) Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Pl Mol Biol Rep 17:74

    Article  Google Scholar 

  • Kiselev KV, Tyunin AP, Zhuravlev YN (2013) Involvement of DNA methylation in the regulation of STS10 gene expression in Vitis amurensis. Planta 237(4):933–941

    Article  CAS  Google Scholar 

  • Körper S, Wink M, Fink RH (1998) Differential effects of alkaloids on sodium currents of isolated single skeletal muscle fibers. FEBS Lett 436(2):251–255

    Article  Google Scholar 

  • Kumari R, Yadav G, Sharma V, Sharma V, Kumar S (2013) Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus. J Genet 92(3):499–511

    Article  CAS  Google Scholar 

  • Liscombe DK, O’Connor SE (2011) A virus-induced gene silencing approach to understanding alkaloid metabolism in Catharanthus roseus. Phytochemistry 72(16):1969–1977

    Article  CAS  Google Scholar 

  • Madhusudan L (2015) Importance of epigenetics in plants. J Bioeng Biomed Sci 5:2

    Google Scholar 

  • Magnotta M, Murata J, Chen J, De Luca V (2007) Expression of deacetylvindoline-4-O-acetyltransferase in Catharanthus roseus hairy roots. Phytochemistry 68(14):1922–1931

    Article  CAS  Google Scholar 

  • Makhzoum A, Petit-Paly G, St Pierre B, Bernards MA (2011) Functional analysis of the DAT gene promoter using transient Catharanthus roseus and stable Nicotiana tabacum transformation systems. Plant Cell Rep 30(7):1173–1182

    Article  CAS  Google Scholar 

  • Mishra P, Kumar S (2000) Emergence of periwinkle Catharanthus roseus as a model system for molecular biology of alkaloids: phytochemistry, pharmacology, plant biology and in vivo and in vitro cultivation. J Med Aromat Plant Sci 22:306–337

    CAS  Google Scholar 

  • Morales-Ruiz T, Ortega-Galisteo AP, Ponferrada-Marín MI, Martínez-Macías MI, Ariza RR, Roldán-Arjona T (2006) DEMETER and REPRESSOR OF SILENCING 1 encode 5-methylcytosine DNA glycosylases. Proc Natl Acad Sci USA 103(18):6853–6858

    Article  CAS  Google Scholar 

  • Murata J, Roepke J, Gordon H, De Luca V (2008) The leaf epidermome of Catharanthus roseus reveals its biochemical specialization. Plant Cell 20(3):524–542

    Article  CAS  Google Scholar 

  • Pandey SS, Singh S, Babu CS, Shanker K, Srivastava NK, Shukla AK, Kalra A (2016) Fungal endophytes of Catharanthus roseus enhance vindoline content by modulating structural and regulatory genes related to terpenoid indole alkaloid biosynthesis. Sci Rep 6:265–283

    Google Scholar 

  • Parashar G, Parashar NCh, Capalash N (2017) – (-) Menthol induces reversal of promoter hypermethylation and associated up-regulation of the FANCF gene in the SiHa cell line. Asian Pac J Cancer Prev 18(5):1365–1370

    PubMed  PubMed Central  Google Scholar 

  • Pasquali G, Erven AS, Ouwerkerk PB, Menke FL, Memelink J (1999) The promoter of the strictosidine synthase gene from periwinkle confers elicitor-inducible expression in transgenic tobacco and binds nuclear factors GT-1 and GBF. Plant Mol Biol 39(6):1299–1310

    Article  CAS  Google Scholar 

  • Power R, Kurz WG, De Luca V (1990) Purification and characterization of acetylcoenzyme A: deacetylvindoline 4-O-acetyltransferase from Catharanthus roseus. Arch Biochem Biophys 279(2):370–376

    Article  CAS  Google Scholar 

  • Shukla AK, Shasany AK, Gupta MM, Khanuja SP (2006) Transcriptome analysis in Catharanthus roseus leaves and roots for comparative terpenoid indole alkaloid profiles. J Exp Bot 57(14):3921–3932

    Article  CAS  Google Scholar 

  • Singh V, Sharma P, Capalash N (2013) DNA methyltransferase-1 inhibitors as epigenetic therapy for cancer. Curr Cancer Drug Targets 13(4):379–399

    Article  CAS  Google Scholar 

  • St-Pierre B, Vazquez-Flota FA, De Luca V (1999) Multi cellular compartmentation of Catharanthus roseus alkaloid biosynthesis predicts intercellular translocation of a pathway intermediate. Plant Cell 11:887–900

    Article  CAS  Google Scholar 

  • Tyunin AP, Kiselev KV, Zhuravlev YN (2012) Effects of 5-azacytidine induced DNA demethylation on methyltransferase gene expression and resveratrol production in cell cultures of Vitis amurensis. Plant Cell Tiss Organ Cult 111:91–100

    Article  CAS  Google Scholar 

  • Van der Fits L, Memelink J (2000) ORCA3, a jasmonate responsive transcriptional regulator of plant primary and secondary metabolism. Science 289:295–297

    Article  Google Scholar 

  • Van Der Heijden R, Jacobs DI, Snoeijer W, Hallard D, Verpoorte R (2004) The Catharanthus alkaloids: pharmacognosy and biotechnology. Curr Med Chem 11(5):607–628

    Article  Google Scholar 

  • Verpoorte R, Van der Heijden R, Moreno PRH (1997) Biosynthesis of terpenoid indole alkaloids in Catharanthus roseus cells. In: Cordell GA (ed) The alkaloids, vol 49. Academic Press, San Diego, pp 221–299

    Google Scholar 

  • Wang Q, Yuan F, Pan Q, Li M, Wang G, Zhao J, Tang K (2010) Isolation and functional analysis of the Catharanthus roseus deacetylvindoline-4-O-acetyltransferase gene promoter. Plant Cell Rep 29(2):185–192

    Article  Google Scholar 

Download references

Acknowledgements

Dr. Nidarshana Chaturvedi Parashar duly acknowledge the funding support provided by Department of Biotechnology, Government of India under DBT-RA program.

Author information

Author notes

  1. Nidarshana Chaturvedi Parashar & Gaurav Parashar

    Present address: Department of Biotechnology, Maharishi Markandeshwar (Deemed to be University), Mullana, Ambala, Haryana, 133207, India

Authors and Affiliations

  1. Department of Biochemistry, Panjab University, Basic Medical Sciences Block II, Sector 25, Chandigarh, 160014, India

    Nidarshana Chaturvedi Parashar & Rajat Sandhir

  2. Laboratory of Epigenetics and Diseases, Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, S.A.S. Nagar (Mohali), Punjab, 160062, India

    Gaurav Parashar

  3. Department of Botany, Panjab University, Chandigarh, 160014, India

    Harsh Nayyar

Authors
  1. Nidarshana Chaturvedi Parashar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gaurav Parashar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Harsh Nayyar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rajat Sandhir
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rajat Sandhir.

Ethics declarations

Conflict of interest

NCP, GP, HN and RS declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Parashar, N.C., Parashar, G., Nayyar, H. et al. Differential DNA methylation in regulation of deacetylvindoline-4-O-acetyl transferase (DAT) gene in Catharanthus roseus. J. Plant Biochem. Biotechnol. 30, 326–335 (2021). https://doi.org/10.1007/s13562-020-00592-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13562-020-00592-7

Keywords

Search

Navigation