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Direct somatic embryogenesis and encapsulation of somatic embryos for in vitro conservation of Bacopa monnieri (L.) Wettst

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Abstract

Direct somatic embryogenesis and shoot organogenesis were achieved from leaf explants excised from microshoots of Bacopa monnieri cultured on Murashige and Skoog medium containing N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of explants differentiated somatic embryos and shoot buds on MS medium supplemented with 12.5 µM BA and 1 µM 2,4-D. The frequency of explants differentiating somatic embryos decreased with increasing concentration of 2,4-D. Light and scanning electron microscopy revealed direct differentiation of somatic embryos and shoot buds from explants, and various developmental stages of the somatic embryos were observed. Somatic embryos and apical shoot tips were encapsulated in sodium alginate gel to produce synthetic seeds. The storage of synthetic seeds produced by encapsulation was studied at 4 and 25 °C (room temperature) for a period of 140 days. Encapsulated somatic embryos were found to retain viability after 140 days of storage at both temperatures, whereas encapsulated apical shoot buds failed to germinate even after 40 days when stored at 4 °C. The viability of synthetic seeds was higher when stored at 25 °C. All amplified markers scored by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) were monomorphic for all the plants produced from synthetic seeds following different periods of storage, thus establishing the clonal fidelity of propagated plantlets.

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Acknowledgments

Amanpreet Kaur is thankful to University Grants Commission (UGC), New Delhi for the award of Maulana Azad National Fellowship for minority students. TIFAC-CORE is thanked for providing facilities to carry out research work.

Author contributions

The work on somatic embryogenesis was carried out by Barkha Khilwani and on synthetic seed was carried out by Ritika Ranjan and Amanpreet Kaur. Amanpreet also wrote an initial draft of the manuscript. Anil Kumar designed all the experiments, corrected the finalized the manuscript.

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Authors and Affiliations

  1. Department of Biotechnology, TIFAC-CORE Building, Thapar University, Patiala, 147001, India

    Barkha Khilwani, Amanpreet Kaur, Ritika Ranjan & Anil Kumar

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  1. Barkha Khilwani
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  2. Amanpreet Kaur
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  3. Ritika Ranjan
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  4. Anil Kumar
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Correspondence to Anil Kumar.

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Khilwani, B., Kaur, A., Ranjan, R. et al. Direct somatic embryogenesis and encapsulation of somatic embryos for in vitro conservation of Bacopa monnieri (L.) Wettst. Plant Cell Tiss Organ Cult 127, 433–442 (2016). https://doi.org/10.1007/s11240-016-1067-5

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  • DOI: https://doi.org/10.1007/s11240-016-1067-5

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