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Encapsulation, short-term storage, conservation and molecular analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum kilimandscharicum Guerke

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Abstract

This article demonstrates the plantlet regeneration from alginate-encapsulated shoot tips of Ocimum kilimandscharicum Guerke. The best gel complexation for encapsulation of shoot tips was achieved using 3 % sodium alginate and 75 mM calcium chloride. Re-growth ability of encapsulated shoot tips explant was evaluated after storage at 4 and 25 °C for 30, 45, 60, 75 and 90 days. Preservation at 25 °C being significantly more resistant compared to that stored in 4 °C. Maximum shoot re-growth (83.33 %) was recorded on 0.7 % agar solidified full strength MS basal medium supplemented with 1.0 mg/l BA. Micro shoots, recovered from encapsulated shoot tips (capsule) were best rooted on ½ strength MS medium containing 1.5 mg/l IBA. Complete plantlets (with shoot and root) were transferred to plastic pots containing soil: vermiculite (1:1) showing 79.53 % survival rate in field condition. The genetic stability of synseed-derived plantlets (following 4 and 25 °C of storage) was assessed and compared with mother plant using two PCR based molecular markers namely random amplified polymorphic DNA and inter-simple sequence repeat. Both types of markers revealed high degree of monomorphism, similar to those of the mother plants.

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Acknowledgments

We are thankful to anonymous reviewers for their valuable suggestions. First author (S. Saha) wish to acknowledge the support of the University Grants Commission (UGC), New Delhi for the award of Post Doctoral Fellowship. Financial assistance provided by University Grant Commission (UGC) New Delhi, is gratefully acknowledged. We also thank to Department of Science and Technology (DST-FIST programme), Govt. of India, Department of Botany, University of Kalyani for Instrumental facilities.

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Authors and Affiliations

  1. Cytogenetics and Plant Biotechnology Research Unit, Department of Botany, University of Kalyani, Kalyani, Nadia, 741235, West Bengal, India

    Soumen Saha & Parthadeb Ghosh

  2. Microbiology Research Laboratory, Department of Botany, University of Kalyani, Kalyani, Nadia, 741235, West Bengal, India

    Chandan Sengupta

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  1. Soumen Saha
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  2. Chandan Sengupta
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Correspondence to Parthadeb Ghosh.

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Saha, S., Sengupta, C. & Ghosh, P. Encapsulation, short-term storage, conservation and molecular analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum kilimandscharicum Guerke. Plant Cell Tiss Organ Cult 120, 519–530 (2015). https://doi.org/10.1007/s11240-014-0618-x

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  • DOI: https://doi.org/10.1007/s11240-014-0618-x

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