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Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa

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Abstract

In vitro grown microshoots of Picrrhiza kurrooa were encapsulated in the alginate beads. Regrowth of encapsulated microshoots, using alginate encapsulation, of P. kurrooa reached 89.33% following 3 months of storage. Amongst developing plantlets, 42.66% exhibited formation of multiple shoots at the onset of regrowth and 21.43% demonstrated simultaneous formation of shoots and roots. Healthy root formation was observed in plantlets following 2 weeks of their transfer to half-strength Murashige and Skoog medium containing 1 μM α-naphthalene acetic acid. Plants were transplanted to the greenhouse in three batches with 95% frequency of survival. The genetic fidelity of P. kurrooa plants growing out after storage in encapsulated form was ascertained by random amplified polymorphic DNA (RAPD) analysis. Molecular analysis of randomly selected plants from each batch was conducted using 45 random decamer primers. Of 45 primes tested, 14 produced scorable amplified products. Total 68 bands were observed amongst them 7.35% bands were polymorphic. Cluster analysis of the RAPD profile revealed an average similarity coefficient of 0.966 thus confirming genetic stability of plants derived from encapsulated microshoots following 3 months of storage.

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Abbreviations

MS:

Murashige and Skoog medium

NAA:

α-naphthalene acetic acid

UPGMA:

Unweighted pair-group method with arithmetic mean

RAPD:

Random Amplified Polymorphic DNA

References

  • Agnihotri RK, Mishra J, Nandi SK (2009) Improving in vitro shoot multiplication and rooting and subsequent field evaluation of genetically uniform regenerants of Dendrocalamus hamiltonii. Acta Physiol Plant 31:961–967

    Article  CAS  Google Scholar 

  • Anand Y, Bansal YK (2002) Synthetic seeds: a novel approach of in vitro plantlet formation in vasaka (Adhatoda vasica. Nees). Plant Biotech 19:159–162

    CAS  Google Scholar 

  • Are H, Jaiswal U, Jaiswal VS (2000) Synthetic seed: prospects and limitations. Curr Sci 78:1438–1444

    Google Scholar 

  • Aronen TS, Krajnakova J, Haggman HM, Ryynanen LA (1999) Genetic fidelity of cryopreserved embryogenic cultures of open-pollinated Abis cephalonica. Plant Sci 142:163–172

    Article  CAS  Google Scholar 

  • Ashmore SE (1997) Current in vitro conservation techniques. In: Engelmann F (ed) Status report on the development and application of in vitro techniques for the conservation and use of plant genetic resources. IPGRI, Rome, pp 5–18

    Google Scholar 

  • Bekheet SA, Taha HS, Saker MM, Solliman ME (2007) Application of cryopreservation technique for in vitro grown date palm (Phoenix dactylifera L.) cultures. J Appl Sci Res 3:853–866

    Google Scholar 

  • Boase MR, Bradley JM, Borst NK (1998) An improved method for transformation of regal Pelargonium (Pelargonium × domesticum Dubonnet) by Agrobacterium tumefaciens. Plant Sci 139:59–69

    Article  CAS  Google Scholar 

  • Borner A (2006) Preservation of plant genetic resources in the biotechnology era. Biotechnology J 1:1393–1404

    Article  Google Scholar 

  • Chandel KPS, Chaudhury R (1995) Desiccation and freezing sensitivity in recalcitrant seeds of tea, cocoa and jackfruit. Ann Bot 76:443–450

    Article  Google Scholar 

  • Dehmer K (2005) Identification of genetic diversity as a precondition of efficient preservation in genebanks. Schr Genet Resour 24:1–6

    Google Scholar 

  • Divakaran M, Babu KN, Pter KV (2006) Conservation of vanilla species in vitro. Sci Hortic 110:175–180

    Article  CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bull 19:11–15

    Google Scholar 

  • Gangopadhyay G, Bandyopadhyay T, Poddar R, Gangopadhyay SB, Mukherjee KK (2005) Encapsulation of pineapple microshoots in alginate beads for temporary storage. Curr Sci 88:972–977

    CAS  Google Scholar 

  • Hirai D, Sakai A (2000) Cryopreservation of in vitro-grown meristems of potato (Solanum tuberosum L.) by encapsulation- verification. In: Engelmann F, Takagi H (eds) Cryopreservation of tropical plant germplasm, current research progress and application. JIRCAS-IPGRI, Rome, pp 205–211

    Google Scholar 

  • Hussain A (1984) Conservation of genetic resources of medicinal plants in India. In: Jain SK (ed) Conservation of tropical plant resources. Botanical Survey of India, Howrah, pp 110–117

    Google Scholar 

  • Jokipii S, Ryynanenb L, Kallioc PT, Aronenb T, Haggman H (2004) A cryopreservation method maintaining the genetic fidelity of a model forest tree, Populus tremula L. × Populus tremuloides Michx. Plant Sci 166:799–806

    Article  CAS  Google Scholar 

  • Kirtikar KR, Basu BD (1984) Indian medicinal plants, vol III. Bishen Singh Mahendra Pal Singh, Dehradun, pp 1824–1826

    Google Scholar 

  • Lata H, Chandra S, Khan IA, Elsohly MA (2009) Propagation through enginate encapsulation of axillary buds of Cannabis sativa L.-an important medicinal plant. Physiol Mol Biol Plants 15:79–86

    Article  CAS  Google Scholar 

  • Mandal J, Patnaik S, Chand PK (2000) Alginate encapsulationof axillary buds of Osimum americanum L. (hoary basil), O. Basilicum L. (sweet basil), O. Gratissimum L. (shrubby basil), and O. Sanctum L. (sacred basil). In Vitro Cell Dev Biol Plant 36:287–292

    CAS  Google Scholar 

  • Mandal AB, Thomas VA, Elanchezhian R (2007) RAPD pattern of Costus speciosus Koen ex. Retz., an important medicinal plant from the Andaman and Nicobar Islands. Curr Sci 93:369–373

    CAS  Google Scholar 

  • Manjkhola S, Dhar U, Joshi M (2005) Organogenesis, embryogenesis and synthetic seed production in Arnebia euchorma—a critically endangered medicinal plant of the Himalaya. In Vitro Cell Dev Biol Plant 41:244–248

    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 

  • Narula A, Kumar S, Srivastava PS (2007) Genetic fidelity of in vitro regenerants, encapsulation of shoot tips and high diosgenin content in Dioscorea bulbifera L., a potential alternative source of diosgenin. Biotechnol Lett 29:623–629

    Article  CAS  PubMed  Google Scholar 

  • Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Nat Acad Sci USA 76:5269–5273

    Article  CAS  PubMed  Google Scholar 

  • Nyende AB, Schittenhelm S, Wagner GM, Greef JM (2003) Production, storability and regeneration of shoot tip of potato (Solanum tuberosum L.) encapsulated in calcium alginate hollow beads. In Vitro Cell Dev Biol Plant 39:540–544

    Article  Google Scholar 

  • Rao NK (2004) Plant genetic resources: advancing conservation and use through biotechnology. Afr J Biotechnol 3:136–145

    Google Scholar 

  • Ray A, Bhattacharyaa S (2008) Storage and plant regeneration from encapsulated shoot tips of Rauvolfia serpentine-an effective way of conservation and mass propagation. S Afr J Bot 74:776–779

    Article  CAS  Google Scholar 

  • Samant SS, Dhar U, Palni LMS (1998) Medicinal plants of Indian Himalaya: diversity, distribution and potential value. Gyanodaya Prakashan, Nainital, p 163

    Google Scholar 

  • Scocchi A, Faloci M, Medina R, Olmos S, Mroginski L (2004) Plant recovery of cryopreserved apical meristem-tips of Melia azedarach L. Using encapsulation/dehydration and assessment of their genetic stability. Euphytica 135:29–38

    Article  CAS  Google Scholar 

  • Shrivastava V, Khan SA, Banerjee S (2009) An evaluation of genetic fidelity of encapsulated microshoots of the medicinal plant: Cineraria maritime following 6 months of storage. Plant Cell Tiss Organ Cult 29:623–629

    Google Scholar 

  • Singh AK, Sharma M, Varshney R, Agarwal SS, Bansal KC (2006a) Plant regeneration from alginate to encapsulated shoot tips of Phyllanthus amarus Schum and Thonn, a medicinally important plant species. In Vitro Cell Dev Biol Plant 42:109–113

    CAS  Google Scholar 

  • Singh AK, Varshney R, Sharma M, Agarwal SS, Bansal KC (2006b) Regeneration of plants from alginate to encapsulated shoot tips of Withania somnifera (L.) Dunal, a medicinally important plant species. J Plant Physiol 163:220–223

    Article  CAS  PubMed  Google Scholar 

  • Standardi A, Piccioni E (1998) Recent perspective on synthetic seed technology using nonembryogenic in vitro-derived explants. Int J Plant Sci 159:968–978

    Google Scholar 

  • Thakur RS, Puri HS, Hussain A (1989) Major medicinal plants of India. CIMAP, Lucknow, pp 404–407

    Google Scholar 

  • Williams K, Kubelik AR, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitary primers are useful as genetic markers. Nucleic Acid Res 18:1631–1635

    Article  Google Scholar 

  • Withers LA (1983) Germplasm storage in plant biotechnology. In: Mantell SH, Smith H (eds) Plant biotechnology. Cambridge University Press, UK, pp 187–218

    Google Scholar 

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Acknowledgments

Author thanks to Ms Himani Bhandari and Mr. Sumit Purohit for their help and cooperation.

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Authors and Affiliations

  1. GB Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora, Uttarakhand, 263 643, India

    Janhvi Mishra, Mahendra Singh, L. M. S. Palni & S. K. Nandi

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  1. Janhvi Mishra
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  2. Mahendra Singh
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  3. L. M. S. Palni
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  4. S. K. Nandi
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Correspondence to Janhvi Mishra.

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Mishra, J., Singh, M., Palni, L.M.S. et al. Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa . Plant Cell Tiss Organ Cult 104, 181–186 (2011). https://doi.org/10.1007/s11240-010-9816-3

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  • DOI: https://doi.org/10.1007/s11240-010-9816-3

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