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High-frequency clonal propagation of Curcuma angustifolia ensuring genetic fidelity of micropropagated plants

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Abstract

A high-frequency clonal propagation protocol was developed for Curcuma angustifolia Roxb., a high valued traditional medicinal plant. Axillary bud explants of C. angustifolia were explanted on Murashige and Skoog (MS) medium fortified with 4.4–22.2 µM 6-benzyladenine (BA), 2.9–5.7 µM indole-3-acetic acid (IAA), 2.3–23.2 µM kinetin (Kin), 2.7–5.4 µM naphthalene acetic acid (NAA) and 67.8-271.5 µM adenine sulphate (Ads) in different combinations. The maximum number of shoots per explants (14.1 ± 0.55) and roots per shoot (7.6 ± 0.47) was achieved on media containing 13.3 µM BA, 5.7 µM IAA and 135.7 µM Ads. Stability in phytomedicinal yield potential of micropropagated plants was assessed through GC–MS and HPTLC. Gas chromatogram of essential oil of conventional and micropropagated plants of C. angustifolia had similar essential oil profile. HPTLC analysis of rhizome extracts of in vitro and field grown plants revealed no significant differences in the fingerprint pattern and in curcumin content. Genetic integrity of in vitro and field grown derived plants were evaluated with inter-simple sequence repeat (ISSR) primers and flow cytometry using Glycine max as an internal standard. A total of 1260 well resolved bands were generated by 12 ISSR primers showing monomorphic banding patterns across all plants analyzed. The mean 2C DNA content of conventionally and micropropagated plant was estimated to be 2.26 pg and 2.31 pg, respectively. As no somaclonal variations were detected in tissue culture plantlets, the present micropropagation protocol could be applied for in vitro conservation and large-scale production of C. angustifolia.

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Acknowledgements

The authors are grateful to Dr. S.C. Si, Dean, Centre of Biotechnology and Dr. M.R. Nayak, President, Siksha ‘O’ Anusandhan University for providing facilities and encouragement throughout.

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Authors and Affiliations

  1. Centre for Biotechnology, Siksha ‘O’ Anusandhan (Deemed to be University), Bhubaneswar, Odisha, India

    Sudipta Jena, Asit Ray, Ambika Sahoo, Suprava Sahoo, Basudeba Kar & Sanghamitra Nayak

  2. Taxonomy and Conservation Division, Regional Plant Resource Centre, Bhubaneswar, Odisha, India

    Pratap Chandra Panda

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  1. Sudipta Jena
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  2. Asit Ray
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  3. Ambika Sahoo
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  4. Suprava Sahoo
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  5. Basudeba Kar
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  6. Pratap Chandra Panda
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  7. Sanghamitra Nayak
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Correspondence to Sanghamitra Nayak.

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Communicated by Nokwanda Pearl Makunga.

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Jena, S., Ray, A., Sahoo, A. et al. High-frequency clonal propagation of Curcuma angustifolia ensuring genetic fidelity of micropropagated plants. Plant Cell Tiss Organ Cult 135, 473–486 (2018). https://doi.org/10.1007/s11240-018-1480-z

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