Skip to main content
Log in

Influence of Incubation Period on Callus Tissues for Plant Regeneration in Ophiorrhiza pectinata Arn. Through Somatic Embryogenesis

  • Research Article
  • Published:
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Ophiorrhiza pectinata Arn. is an important medicinal plant reported to contain camptothecin which is used for the treatment of different types of cancers. The present study describes an efficient plant regeneration system through somatic embryogenesis. Embryogenic callus tissues raised from aseptic leaf explants were used for inducing somatic embryos under incubation of callus tissues at varying periods from 10 to 60 days. It was noticed that callus tissues incubated between 30 and 50 days in 0.5 mg l−1 2,4-dichlorophenoxy acetic acid were responded positively in half-strength Murashige and Skoog liquid media supplemented with 3 mg l−1 indole-3-acetic acid and started differentiation of globular structures in 15–20 days. Maximum differentiation of embryos was obtained in callus tissues incubated for 40 days. Clumps of embryos at different stages got differentiated into plantlets in MS media supplemented with 6-benzyl aminopurine (0.2 mg l−1) and indole-3-acetic acid (0.1 mg l−1) and produced an average of 47.92 ± 5.33 plants from 1.0 to 1.5 g callus tissues. The regenerated plantlets were successfully deflasked, hardened and established in paper cups filled with coarse sand and recorded more than 90% establishment under shade net house (50% shade) condition with adequate watering. The method of plant regeneration system developed would be useful for raising large number of plants and production of bioactive compounds.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Deb DB, Mondal DC (1997) Taxonomic revision of the genus Ophiorrhiza L. (Rubeaceae) in Indian subcontinent. Nelumbo India 39:1–48

    Google Scholar 

  2. Joseph G, Joseph JP (2009) Rediscovery of Ophiorrhiza caudata (Rubiaceae) from the Western Ghats of Kerala. Rheedea 19(1):45–46

    Google Scholar 

  3. Rajan R, Varghese SC, Kurup R, Gopalakrishnan R, Venkataraman R, Satheeshkumar K, Baby S (2013) Search for camptothecin-yielding Ophiorrhiza species from southern Western Ghats in India: a HPTLC-densitometry study. Ind Crops Prod 43:472–476. https://doi.org/10.1016/j.indcrop.2012.07.054

    Article  CAS  Google Scholar 

  4. Kirtikar KR, Basu BD (1975) Indian medicinal plant. Bishen Singh Mahendrapal Singh, New Delhi

    Google Scholar 

  5. Baiju EC, Asha A, Sneha J, Athira T (2018) In vitro antioxidant and antibacterial activities of two Ophiorrhiza species. Int J Green Herb Chem 7(1):033–038. https://doi.org/10.24214/IJGHC/HC/7/1/03338

    Article  CAS  Google Scholar 

  6. Wall ME, Wani MC, Cook CE, Palmer AT, McPhail AT, Sim GA (1966) Plant antitumor agents. I. The isolation and structure of camptothecin, a novel alkaloidal leukemia and tumor inhibitor from Camptotheca acuminata 1,2. J Am Chem Soc 88:3888–3890. https://doi.org/10.1021/ja00968a057

    Article  CAS  Google Scholar 

  7. Garcia Carbonero R, Supko JG (2002) Current perspectives on the clinical experience, pharmacology, and continued development of the camptothecin. Clin Cancer Res 8:641–661

    CAS  PubMed  Google Scholar 

  8. Redinbo MR, Stewart L, Kuhn P, Champoux JJ, Hol WG (1998) Crystal structures of human topoisomerase I in covalent and non-covalent complexes with DNA. Science 279:1504–1513. https://doi.org/10.1126/science.279.5356.1504

    Article  CAS  PubMed  Google Scholar 

  9. Hsiang YH, Hertzberg R, Hecht S, Liu LF (1985) Camptothecin induce protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem 260:14873–14878

    CAS  PubMed  Google Scholar 

  10. Silja PK, Gisha GP, Satheeshkumar K (2014) Enhanced plumbagin accumulation in embryogenic cell suspension cultures of Plumbago rosea L. following elicitation. Plant Cell Tissue Organ Cult 119(3):469–477. https://doi.org/10.1007/s11240-014-0547-8

    Article  CAS  Google Scholar 

  11. Vasil IK (1984) Cell culture and somatic cell genetics of plants. Academic Press, Inc., Cambridge

    Google Scholar 

  12. Jiménez VM (2005) Involvement of plant hormones and plant growth regulators on invitro somatic embryogenesis. Plant Growth Regul 47(2–3):91–110. https://doi.org/10.1007/s10725-005-3478-x

    Article  CAS  Google Scholar 

  13. Pagnussat GC, Alandete-Saez M, Bowman JL, Sundaresan V (2009) Auxin dependent patterning and gamete specification in the Arabidopsis female gametophyte. Science 324(5935):1684–1689. https://doi.org/10.1126/science.1167324

    Article  CAS  PubMed  Google Scholar 

  14. Papori Phukan Borpuzari and Mina Borthakur (2016) Effect of plant growth regulators and explants sources on somatic embryogenesis of matured tissue of the anticancerous medicinal plant Plumbago rosea. J Med Plant Stud 4(5):165–170

    Google Scholar 

  15. Raghavan V (2003) One hundred years of zygotic embryo culture investigations. In Vitro Cell Dev Biol Plant 39(5):437–442. https://doi.org/10.1079/IVP2003436

    Article  Google Scholar 

  16. Choi YE, Ko SK, Lee KS, Yoon ES (2002) Production of plantlets of Eleutherococcus sessiliflorus via somatic embryogenesis and successful transfer to soil. Plant Cell Tissue Organ Cult 69(2):201–204. https://doi.org/10.1023/A:1015290718273

    Article  CAS  Google Scholar 

  17. Kumar HGA, Murthy HN, Paek KY (2002) Somatic embryogenesis and plant regeneration in Gymnema sylvestre. Plant Cell Tissue Organ Cult 71:85–88. https://doi.org/10.1023/A:1016529322046

    Article  Google Scholar 

  18. Vikrant Rashid A (2003) Somatic embryogenesis from mesocotyl and leaf-base segments of Paspalum scrobiculatum L., a minor millet. In Vitro Cell Dev Biol Plant 39:485–489. https://doi.org/10.1079/IVP2003457

    Article  Google Scholar 

  19. Williams EG, Maheswaran G (1986) Somatic embryogenesis: factors influencing coordinated behaviour of cells as an embryogenic group. Ann Bot 57:443–462. https://doi.org/10.1093/oxfordjournals.aob.a087127

    Article  Google Scholar 

  20. Sujhatha M, Reddy TP (1998) Differential cytokinin effects on the stimulation of in vitro shoot proliferation from meristematic explants of castor (Ricinus communis L.). Plant Cell Rep 17:561–566. https://doi.org/10.1007/s002990050442

    Article  Google Scholar 

  21. Nagesh KS, Shanthamma C, Pullaiah T (2010) Somatic embryogenesis and plant regeneration from callus cultures of Curculigo orchioides Gaertn. Indian J Biotechnol 9:408–413. http://hdl.handle.net/123456789/10440

  22. Raghavan V (2005) Control of leaf formation and somatic embryogenesis in cultured zygotic embryos of Arabidopsis thaliana by 2,4-dichlorophenoxyacetic acid (2,4-D). Int J Plant Sci 166(4):575–588. https://doi.org/10.1086/430336

    Article  CAS  Google Scholar 

  23. Lo Schiavo F, Pitto L, Giuliano G, Torti G, Nuti-Ronchi V, Marazziti D, Veraga R, Orselli S, Terzi M (1989) DNA methylation of embryogenic carrot cell cultures and its variations as caused by maturation, differentiation, hormones and hypomethylation drugs. Theor Appl Genet 77:325. https://doi.org/10.1007/BF00305823

    Article  CAS  Google Scholar 

  24. Ammirato PV (1986) Organizational events during somatic embryogenesis. In: Green CE, Somers DA, Hackett WP, Biesboer DD (eds) Plant biology, vol 3. Alan R Liss, New York, pp 57–81

    Google Scholar 

  25. Gutiérrez-Mora A, González-Gutiérre AG, Rodríguez-Garay B, Ascencio-Cabral A, Li-Wei L (2012) Plant somatic embryogenesis: some useful considerations. In: Sato K-I (ed) Embryogenesis. InTech, London, pp 229–248

    Google Scholar 

  26. Lakshmanan P, Taji A (2000) Somatic embryogenesis in leguminous plants. Plant Biol 2(2):136–148. https://doi.org/10.1055/s-2000-9159

    Article  CAS  Google Scholar 

  27. Martin KP, Beegum AS, Zhang CL, Slater A, Madhusoodanan PV (2007) In vitro propagation of Ophiorrhiza prostrata through somatic embryogenesis. Biol Plant 51(4):769–772. https://doi.org/10.1007/s10535-007-0157-y

    Article  CAS  Google Scholar 

  28. Kim SW, Song NH, Jung KH, Kwak SS, Liu JR (1994) High frequency plant regeneration from anther-derived cell suspension cultures via somatic embryogenesis in Catharanthus roseus. Plant Cell Rep 13(6):319–322. https://doi.org/10.1007/BF00232629

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank to the Director of the Institute for providing facilities. They also acknowledge Kerala State Council for Science Technology and Environment (KSCSTE) and University of Kerala for financial support as Ph.D. Fellowship (Order Number is No. Ac.EVI(1)/55039).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to K. Satheeshkumar.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest to publish this manuscript.

Additional information

Significance statement The protocol developed in the present investigation could be a novel method of plant regeneration in Ophiorrhiza pectinata Arn. through somatic embryogenesis.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Midhu, C.K., Hima, S., Binoy, J. et al. Influence of Incubation Period on Callus Tissues for Plant Regeneration in Ophiorrhiza pectinata Arn. Through Somatic Embryogenesis. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 1439–1446 (2019). https://doi.org/10.1007/s40011-018-01061-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40011-018-01061-x

Keywords

Navigation