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Terpene profiling, transcriptome analysis and characterization of cis-β-terpineol synthase from Ocimum

  • Atul Anand
  • Ramesha H. Jayaramaiah
  • Supriya D. Beedkar
  • Bhushan B. Dholakia
  • Santosh G. Lavhale
  • Sachin A. Punekar
  • Wasudeo N. Gade
  • Hirekodathakallu V. Thulasiram
  • Ashok P. Giri
Research Article
  • 91 Downloads

Abstract

Ocimum species produces a varied mix of different metabolites that imparts immense medicinal properties. To explore this chemo-diversity, we initially carried out metabolite profiling of different tissues of five Ocimum species and identified the major terpenes. This analysis broadly classified these five Ocimum species into two distinct chemotypes namely, phenylpropanoid-rich and terpene-rich. In particular, β-caryophyllene, myrcene, limonene, camphor, borneol and selinene were major terpenes present in these Ocimum species. Subsequently, transcriptomic analysis of pooled RNA samples from different tissues of Ocimum gratissimum, O. tenuiflorum and O. kilimandscharicum identified 38 unique transcripts of terpene synthase (TPS) gene family. Full-length gene cloning, followed by sequencing and phylogenetic analysis of three TPS transcripts were carried out along with their expression in various tissues. Terpenoid metabolite and expression profiling of candidate TPS genes in various tissues of Ocimum species revealed spatial variances. Further, putative TPS contig 19414 (TPS1) was selected to corroborate its role in terpene biosynthesis. Agrobacterium-mediated transient over-expression assay of TPS1 in the leaves of O. kilimandscharicum and subsequent metabolic and gene expression analyses indicated it as a cis-β-terpineol synthase. Overall, present study provided deeper understanding of terpene diversity in Ocimum species and might help in the enhancement of their terpene content through advanced biotechnological approaches.

Keywords

Agro-infiltration assay Metabolite profiling Ocimum Terpenes Terpene synthase Transcriptome 

Notes

Acknowledgements

AA and SGL acknowledge Council of Scientific and Industrial Research (CSIR), New Delhi for SRF while SB acknowledges University Grants Commission, New Delhi, India for Kothari fellowship. The work was funded by CSIR-NCL-IGIB Joint Research program under XII Five Year Plan (BSC0124).

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

12298_2018_612_MOESM1_ESM.doc (36 kb)
Supplementary material 1 (DOC 36 kb)
12298_2018_612_MOESM2_ESM.doc (51 kb)
Supplementary material 2 (DOC 51 kb)
12298_2018_612_MOESM3_ESM.xls (135 kb)
Supplementary material 3 (XLS 135 kb)

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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  • Atul Anand
    • 1
    • 2
  • Ramesha H. Jayaramaiah
    • 1
  • Supriya D. Beedkar
    • 2
    • 3
  • Bhushan B. Dholakia
    • 1
  • Santosh G. Lavhale
    • 1
  • Sachin A. Punekar
    • 4
  • Wasudeo N. Gade
    • 3
  • Hirekodathakallu V. Thulasiram
    • 2
    • 5
  • Ashok P. Giri
    • 1
  1. 1.Plant Molecular Biology Unit, Division of Biochemical SciencesCSIR-National Chemical LaboratoryPuneIndia
  2. 2.Chemical Biology Unit, Division of Organic ChemistryCSIR-National Chemical LaboratoryPuneIndia
  3. 3.Department of BiotechnologySavitribai Phule Pune University (Formerly University of Pune)PuneIndia
  4. 4.Biospheres, EshwariPuneIndia
  5. 5.CSIR-Institute of Genomics and Integrative BiologyNew DelhiIndia

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