Skip to main content
SpringerLink
Log in

Micropropagation of Zingiber rubens and assessment of genetic stability through RAPD and ISSR markers

  • Published:
Biologia Plantarum

Abstract

Protocol was developed for high frequency in vitro multiplication of an endemic species, Zingiber rubens Roxb. The sprouted buds of the rhizomes were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA; 0.5–5.0 mg dm−3), indole-3-acetic acid (IAA; 0.5–2.0 mg dm−3), kinetin (KIN; 1.0–3.0 mg dm−3), naphthaleneacetic acid (NAA; 0.5–1.0 mg dm−3) and adenine sulphate (ADS; 80–100 mg dm−3). MS basal medium supplemented with 3 mg dm−3 BA and 0.5 mg dm−3 IAA was optimum for shoot elongation. The elongated shoots (1–2 cm) were transferred to multiplication medium containing 2 mg dm−3 BA, 1 mg dm−3 IAA and 100 mg dm−3 ADS. The multiplication rate remained unchanged in subsequent subcultures. Upon ex vitro transfer, 85 % of plants survived. Genetic stability of micropropagated clones were periodically evaluated at an interval of 6 months up to 30 months in culture using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis and genetic uniformity in all regenerants was confirmed.

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

Similar content being viewed by others

Abbreviations

ADS:

adenine sulphate

BA:

benzyladenine

IAA:

indoleacetic acid

MS:

Murashige and Skoog medium

NAA:

napthaleneacetic acid

KIN:

kinetin

RAPD:

random amplified polymorphic DNA

ISSR:

inter simple sequence repeats

References

  • Balachandran, S.M., Bhat, S.R., Chandel, K.P.S.: In vitro clonal multiplication of turmeric (Curcuma sp.) and ginger (Zingiber officinale Rosc.). — Plant Cell Rep. 8: 521–524, 1990.

    Article  CAS  Google Scholar 

  • Bhagyalakshmi, N., Singh, N.S.: Meristem culture and micropropagation of a variety of ginger (Zingiber officinale Rosc.) with a high yield of oleoresin. — J. hort. Sci. 63: 321–327, 1988.

    CAS  Google Scholar 

  • Chang, B.K.W., Criley, R.A.: Micropropagation of pink ginger in vitro. — Hort. Sci. 28: 1203–1205, 1993.

    Google Scholar 

  • Munoz, C.M.: Micropropagation of endangered species. — Ecol. mediter. 21: 291–297, 1995.

    Google Scholar 

  • Dai, X., Shi, S., Ye, Y., Fu, Q., Bao, M.: High frequency plant regeneration from cotyledon and hypocotyl explants of ornamental kale. — Biol. Plant. 53: 769–773, 2009.

    Article  Google Scholar 

  • Doyle, J.J., Doyle, J.L.: A rapid DNA isolation procedure for small quantities of fresh leaf tissue. — Phytochem. Bull. 19: 11–15, 1990.

    Google Scholar 

  • Faria, R.I., Illg, R.D.: Micropropagation of Zingiber spectabile Griff. — Sci. Hort. 62: 135–137, 1995.

    Article  Google Scholar 

  • Gonçalves, S., Martins, N., Romano, A.: Micropropagation and conservation of endangered species Plantago algarbiensis and P. almogravensis. — Biol. Plant. 53: 774–778, 2009.

    Article  Google Scholar 

  • Hammerschlag, F.A., Bauchan, G.R., Scorza, R.: Factors influencing in vitro multiplication and rooting of peach cultivars. — Plant Cell Tissue Organ Cult. 8: 235–242, 1987.

    Article  CAS  Google Scholar 

  • Ikeda, I.R., Tambe, M.J.: In vitro subculture application for ginger. — Hort. Sci. 24: 142–143, 1989.

    Google Scholar 

  • Jain, S.K., Prakash, V.: Zingiberaceae in India: phytogeography and endemism. — Rheedea 5: 154–169, 1994.

    Google Scholar 

  • Joshi, P., Dhawan, V.: Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. — Biol. Plant. 51: 22–26, 2007.

    Article  CAS  Google Scholar 

  • Kacker, A., Bhat, S.R., Chandel, K.P.S., Malik, S.K.: Plant regeneration via somatic embryogenesis in ginger. — Plant Cell Tissue Organ Cult. 32: 289–292, 1993.

    Article  Google Scholar 

  • Larkins, P., Scowcroft, W.R.: Somaclonal variation, a novel source of variability from cell cultures for plant improvement. — Theor. appl. Genet. 60: 197–214, 1981.

    Article  Google Scholar 

  • Mallikarjuna, K., Rajendrudu, G.: Rapid in vitro propagation of Holarrhena antidysenterica using seedling cotyledonary nodes. — Biol. Plant. 53: 569–572, 2009.

    Article  CAS  Google Scholar 

  • Martins, M., Sarmento, D., Oliveira, M.M.: Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers. — Plant Cell Rep. 23: 492–496, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Mohanty, S., Panda, M.K., Subudhi, E., Acharya, L., Nayak, S.: Genetic stability of micropropagated ginger derived from axillary bud through cytophotometric and RAPD analysis. — Z Naturforsch. 63c: 747–754, 2008.

    Google Scholar 

  • Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. — Physiol. Plant. 15: 473–497, 1962.

    Article  CAS  Google Scholar 

  • Nayak, S., Sen, S.: Differential resistance of three species of Ornithogalum to polyploidization in vitro. — Nucleus 41: 48–52, 1998.

    Google Scholar 

  • Panda, M.K., Mohanty, S., Subudhi, E., Acharya, L., Nayak, S.: Assessment of genetic stability of micropropagated plants of Curcuma longa L. by cytophotometry and RAPD analysis. — Int. J. Integr. Biol. 1: 189–195, 2007.

    CAS  Google Scholar 

  • Poonasapaya, P., Kraisintu, K.: Micropropagation of Zingiber cassumunar Roxb. — Acta Hort. 344: 557–564, 1993.

    Google Scholar 

  • Rout, G.R., Das, P.: In vitro studies of ginger: a review of recent progress. — In: Goril, J.N., Anand Kumar, P., Singh, V.K. (ed.): Recent Progress in Medicinal Plants. Vol. 4. Biotechnology and Genetic Engineering. Pp. 307–326. Studium Press, Houston 2002.

    Google Scholar 

  • Salvi, N.D., George, L., Eapen, S.: Micropropagation and field evaluation of micro-propagated plants of turmeric. — Plant Cell Tissue Organ Cult. 68: 143–151, 2002.

    Article  Google Scholar 

  • Sharma, T.R., Singh, B.M.: High frequency in vitro multiplication of disease free Zingiber officinale Rosc. — Plant Cell Rep. 17: 68–72, 1997.

    Article  CAS  Google Scholar 

  • Smith, M.K., Hamil, S.D.: Field evaluation of micro-propagated ginger in subtropical Queensland. — Aust. J. exp. Agr. 36: 347–354, 1996.

    Article  Google Scholar 

  • Tyagi, R.K., Agrawal, A., Yusup, A.: Conservation of Zingiber germplasm through in vitro rhizome formation. — Sci. Hort. 108: 210–219, 2006.

    Article  CAS  Google Scholar 

  • Vendrame, W.A., Kochert, G., Wetzstein, H.Y.: AFLP analysis of variation in pecan somatic embryos. — Plant Cell Rep. 18: 853–857, 1999.

    Article  CAS  Google Scholar 

  • Venkatachalam, L., Sreedhar, R.V., Bhagyalakshmi, N.: Genetic analysis of micropropagated and regenerated plantlets of banana as assessed by RAPD and ISSR markers. — In vitro cell. dev. Biol. 43: 267–274, 2007.

    Article  CAS  Google Scholar 

  • Williams, J.G.K., Kubelik, A.R., Liva, K.J., Rafalski, J.A., Tingey, S.V.: DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. — Nucl. Acids Res. 18: 6531–6535, 1990.

    Article  CAS  PubMed  Google Scholar 

  • Zeitkiewicz, E., Rafalski, A., Labuda, D.: Genome finger printing by simple sequence repeat (SSR)-anchored PCR amplification. — Genomics 20: 176–183, 1994.

    Article  Google Scholar 

  • Zucchi, M.I., Arizono, H., Morais, V.I., Fungaro, M.H.P., Vieira, M.L.C.: Genetic instability of sugarcane plants derived from meristem cultures. — Genet. mol Biol. 25: 91–96, 2002.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Dr. S.C. Si, Centre of Biotechnology and Dr. M.R. Nayak, Anusandhan University for providing facilities and encouraging throughout. Financial assistance from Department of Biotechnology, New Delhi, India, is also duly acknowledged.

Author information

Authors and Affiliations

  1. Centre of Biotechnology, Siksha O Anusandhan University, Khandagiri, Bhubaneswar-30, Orissa, India

    S. Mohanty, M. K. Panda & S. Nayak

  2. P.G. Department of Botany, Utkal University, Vanivihar, Bhubaneswar, Orissa, India

    S. Sahoo

Authors
  1. S. Mohanty
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. K. Panda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Sahoo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. Nayak
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. Nayak.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mohanty, S., Panda, M.K., Sahoo, S. et al. Micropropagation of Zingiber rubens and assessment of genetic stability through RAPD and ISSR markers. Biol Plant 55, 16–20 (2011). https://doi.org/10.1007/s10535-011-0002-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-011-0002-1

Additional key words

Search

Navigation