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In vitro and ex vitro evaluation of long-term micropropagated turmeric as analyzed through cytophotometry, phytoconstituents, biochemical and molecular markers

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Abstract

Turmeric (Curcuma longa L.), a high valued medicinal plant, was micropropagated through induction of multiple shoots using latent axillary buds of rhizome. Cytophotometric and random amplified polymorphic DNA (RAPD) as well as inter simple sequence repeats (ISSR) analysis were used to periodically monitor the genetic stability of micropropagated clones of Curcuma longa conserved in vitro up to 7 years at every 6 months interval. A total of eighteen RAPD and eight ISSR primers gave 45,537 distinct and reproducible bands, monomorphic across all 353 plants analyzed. Micropropagated turmeric after being conserved for 7 years in vitro was transplanted into soil in field. Drug yielding potential of tissue culture derived plants was evaluated in field through estimation of phytoconstituents like curcumin and essential oil contents. The result of 2 years of field trial showed that micropropagated turmeric retained stability in all the characteristics examined when compared with the field performance of conventionally propagated plants. Thus long term conservation of an elite genotype of turmeric with epigenetic and genetic stability is significant for stable supply of drug i.e., curcumin and essential oil to the market.

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References

  • Ashmore SE (1997) Status report on the development and application of in vitro techniques for the conservation and use of plant genetic resources. International Plant Genetic Resources Institute, Rome, Italy, p 67

    Google Scholar 

  • Balachandran SM, Bhat SR, Chandel KPS (1990) In vitro clonal multiplication of turmeric (Curcuma longa) and ginger (Zingiber officinale Rosc.). Plant Cell Rep 3:521–524

    Article  Google Scholar 

  • Behura S, Srivastava VK (2004) Essential oils of leaves of curcuma species, J Essent Oil Res

  • Bhatia R, Singh KP, Jhang T, Sharma TR (2009) Assesment of genetic fidelity of micropropagated gerbera plants by ISSR markers. Sci Hort 11(2):208–211

    Article  Google Scholar 

  • Doyle JJ, Doyle JL (1990) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • Goto S, Thakur RC, Ishii K (1998) Determination of genetic stability in long term micropropagated shoots of Pinus thunbergii Parl. Using RAPD markers. Plant Cell Rep 18:193–197

    Article  CAS  Google Scholar 

  • Gould AR (1986) Factors controlling generation of variability in vitro. In: Vasil IK (ed) Cell culture and somatic cell genetics in plants 3, plant regeneration and genetic variability. Academic, Orlando, pp 549–567

    Google Scholar 

  • Guenther E (1972) In: Robert E (ed) The essential oils, vol 1, Krieger Publication Co, Huntington, New York, pp 361–391

  • Hammerschlag FA, Bauchan GR, Scorza R (1987) Factors influencing in vitro multiplication and rooting of peach cultivars. Plant Cell Tissue Organ Cult 8:235–242

    Article  CAS  Google Scholar 

  • Hartmann C, Henry Y, De Buyser J, Aubry C, Rode A (1989) Identification of new mitochondrial genome organizations in wheat plants regenerated from somatic tissue cultures. Theor Appl Genet 77:169–175

    Article  Google Scholar 

  • Hatano K, Kamura K, Shoyama Y, Nishioka I (1988) Clonal propagation of Aconitum carmichaeli by tip tissue culture and alkaloid contents of clonally propagated plants. Planta Med 54:152–154

    Article  PubMed  CAS  Google Scholar 

  • Hussey G (1986) Problems and prospects in the in vitro propagation of herbaceous plants. In: Withers LA, Aldeson PG (eds) Plant tissue culture and its agricultural application. Butterworths, London, pp 113–123

    Google Scholar 

  • Joshi P, Dhawan V (2007) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51(1):22–26

    Article  CAS  Google Scholar 

  • Kuanar A, Mohanty S, Panda MK, Nayak S (2009) Essential oils from leaves of micropropagated turmeric. Curr Sci 96:1166–1167

    Google Scholar 

  • LarKns P, Scowcroft WR (1981) Somaclonal variation, a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214

    Article  Google Scholar 

  • Leela NK, Tava A, Shafi PM, Chempakam B (2002) Chemical composition of essential oils of turmeric (Curcuma longa L.). Acta Pharml 52:137–141

    CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    Google Scholar 

  • Mohanty S, Joshi RK, Subudhi E, Sahoo S, Nayak S (2010) Assessment of genetic stability of micropropagated Curcuma caesia through cytophotometric and molecular analysis. Cytologia 75(1):73–81

    Article  CAS  Google Scholar 

  • Morata-Renau B, Nebauer SG, Arrillaga I, Segura J (2005) Assesment of somaclonal variation in micropropagated shoots of Cedrus:consequences axillary bud breaKng. Tree Genet Gen 1:3–10

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nadgauda RS, Mascarenhas AF, Hendre RR, Jagannathan V (1978) Rapid clonal multiplication of turmeric Curcuma longa L plants by tissue culture. Ind J Exp Biol 16:120–122

    CAS  Google Scholar 

  • Nayak S, Naik PK (2006) Factors affecting in vitro micro rhizome formation and growth in Curcuma longa and improved field performance of micropropagated plants. Sci Asia 32:31–37

    Article  CAS  Google Scholar 

  • Nayak S, Sen S (1991) Cytological and cytophotometrical analysis of direct explants and callus derived plants of Ornthogallum thrysoids Jacq. Cytologia 56:297–302

    Google Scholar 

  • Nayak S, Sen S (1995) Rapid and stable propagation of Ornithgalam umbellatum L. in long term culture. Plant Cell Rep 15:150–153

    Article  CAS  Google Scholar 

  • Nayak S, Sen S (1997) Cytological and cytophotometrical analysis of callus and regereated palants of Ornthogallum virens. Cytobios 91:135–142

    Google Scholar 

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

    Google Scholar 

  • Nayak S, Debta BK, Sahoo S (1996) Rapid propagation of lemongrass (Cymbopogon flexuosus (Nees) Wats) through somatic embryogenesis in vitro plants. Plant Cell Rep 15:367–370

    Article  CAS  Google Scholar 

  • Nayak S, Mohanty S, Subudhi E (2008) Differential synthesis of essential oil in callus derived microshoots of turmeric (Curcuma longa) in vitro, in International conference of Association of plant tissue culture. 12–17th oct 2008, Dalian, China

  • Official Analytical Methods of the American Spice Trade Association (1997) Method No. 1.06. 4th edn. ASHA Press

  • Official Analytical Methods of the American Spice Trade Association (1997b) Method No. 1.09. 4th edn. ASHA Press

  • Orton TJ (1985) Genetic instability during embryogenic cloning of celery. Plant Cell Tissue Organ Cult 4:159–169

    Article  Google Scholar 

  • Panda MK, Mohanty S, Subudhi E, Acharya L, Nayak S (2007) Assessment of genetic stability of micropropagated plants of Curcuma longa L. by cytophotometry and RAPD analysis. Inter J Int Biol 1(3):189–195

    CAS  Google Scholar 

  • Peredo EL, Arroyo-Garcia R, Revilla MA (2009) Epigenetic changes detected in micropropagated hop plants. J Plant Physiol 166(10):1101–1111

    Article  PubMed  CAS  Google Scholar 

  • Potter R, Jones MGK (1991) An assessment of genetic stability of potato in vitro by molecular and phenotypic analysis. Plant Sci 76:239–248

    Article  CAS  Google Scholar 

  • Roses IA (1999) Medicinal plants of the world: chemical constituents, traditional and modern medicinal uses. Humana Press, New Jersey, pp 139–153

    Google Scholar 

  • Rout GR, Das G (2002) An assessment of genetic integrity of micropropagated plants of Plumbago zeylanica by RAPD markers. Biol Plantarum 45(1):27–32

    Article  CAS  Google Scholar 

  • Salvi ND, George L, Eapen S (2001) Plant regeneration from leaf base callus of turmeric and random amplification polymorhic DNA analysis of regenerated plants. Plant Cell Tiss Org Cult 66:113–119

    Article  CAS  Google Scholar 

  • Salvi ND, George L, Eapen S (2002) Micropropagation and field evaluation of micropropagated plants of turmeric. Plant Cell Tiss Org Cult 68:143–151

    Article  Google Scholar 

  • Sharma AK, Sharma A (1980) Chromosome techniques: theory & practice. 3rd edn. Butterworths, London

    Google Scholar 

  • Shenoy VB, Vasil IK (1992) Biochemical and molecular analysis of plants derived from embryogenic tissue cultures of napiergrass (Penisetum purpureum K. Schum.). Theor Appl Genet 83:947–955

    Article  CAS  Google Scholar 

  • Smykal P, Valledor L, Rodriguez R, Griga M (2007) Assesment of genetic and epigenetic stability in long term in vitro shoot culture of pea (Pisum sativum L.). Plant Cell Rep 26:1985–1998

    Article  PubMed  CAS  Google Scholar 

  • Tyagi RK, Yusuf A, Dua P, Agrawal A (2004) In vitro plant regeneration and genotype conservation of eight wild species of Curcuma. Biol Plant 48:129–132

    Article  CAS  Google Scholar 

  • Tyagi RK, Agrawal A, Yusuf A (2006) Conservation of Zingiber germplasm through in vitro rhizome formation. Scientia Hortic 108:210–219

    Article  CAS  Google Scholar 

  • Van’t Hof J (1965) Relationships between mitotic cycle duration, S period duration and the average rate of DNA synthesis in the root meristem cells of several plants. Exp Cell Res 39:48–58

    Article  Google Scholar 

  • Williams JGK, Kubelik AR, Liva KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acid Res 18:6531–6535

    Article  CAS  Google Scholar 

  • Xiaoqiang MA, Gang DR (2006) Metabolic profiling of in vitro micropropagated and conventionally greenhouse grown ginger (Zingiber officinale). Phytochemistry 67(20):2239–2255

    Article  Google Scholar 

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

    Article  Google Scholar 

  • Zucchi MI, Arizono H, Morais VI, Fungaro MHP, Vieira MLC (2002) Genetic instability of sugarcane plants derived from meristem cultures. Genet and Mol Biol 25(1):91–96

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Prof. Dr. Sudam Chandra Si, Dean and Prof. Dr. Manoj Ranjan Nayak, President, Center of Biotechnology, Siksha O Anusandhan University, for providing all facilities.

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Correspondence to Sanghamitra Nayak.

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Nayak, S., Kaur, T., Mohanty, S. et al. In vitro and ex vitro evaluation of long-term micropropagated turmeric as analyzed through cytophotometry, phytoconstituents, biochemical and molecular markers. Plant Growth Regul 64, 91–98 (2011). https://doi.org/10.1007/s10725-010-9541-2

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  • DOI: https://doi.org/10.1007/s10725-010-9541-2

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