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Somatic Embryogenesis and Synthetic Seed Production—a Biotechnological Approach for True-to-Type Propagation and In Vitro Conservation of an Ornamental Bulbaceous Plant Drimiopsis kirkii Baker.

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Abstract

An efficient plant regeneration protocol through indirect somatic embryogenesis pathway via callus had been developed from the leaf explant of an ornamental bulbaceous plant Drimiopsis kirkii. Optimum friable calli were induced on Murashige and Skoog (MS) basal medium supplemented with 3.0 mg/l of 2,4-dichlorophenoxyacetic acid and 1.0 mg/l of α-naphthalene acetic acid (NAA). On subculturing the callus on MS medium supplemented with 2.5 mg/l of thidiazuron (TDZ), 73.3 % of the cultures responded with 20.4 ± 0.3 somatic embryos (SEs) per 500 mg callus at different stages of development after 6 weeks of culture. The highest response of 86.7 % with 28.3 ± 0.5 embryos per 500 mg callus was observed on MS medium supplemented with 2.5 mg/l TDZ and 1.0 mg/l NAA. SEs were encapsulated in calcium alginate beads for the production of synthetic seeds (SSs) and their storability was investigated. The highest SS germination (93.3 %) was observed in 1.0 % sodium alginate followed by 86.7 % germination with 2.5 % sodium alginate. The SSs were stored at three different temperatures (4, 15, and 24 ºC) up to 6 months. The SSs kept at 15 °C showed 64.4 % germinability even after 4 months of storage. Both nonencapsulated and encapsulated SE-derived plants were successfully transferred to soil with 93.3 and 88.3 % survival rate accordingly. Randomly amplified polymorphic DNA (RAPD) analysis revealed that there were no somaclonal variations among the plants produced via somatic embryogenesis and they are true-to-type to their parental plant. These results confirmed the most reliable methods, which can be further used for genetic transformation studies as well as for mass propagation of ornamental D. kirkii at a commercial level.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

EM:

Encapsulation medium

KIN:

Kinetin

MS:

Murashige and Skoog

NAA:

α-Naphthalene acetic acid

PGRs:

Plant growth regulators

RAPD:

Randomly amplified polymorphic DNA

SEs:

Somatic embryos

SSs:

Synthetic seeds

TDZ:

Thidiazuron

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Acknowledgments

SMH acknowledges the Ministry of Minority Affairs (MOMA) and the University Grant Commission (UGC) for providing Maulana Azad National Fellowship (MANF) as well as for the financial support to conduct these studies. Both authors acknowledge Swami Kamalasthananda, Principal, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata (India) for the facilities provided during the present study. Also, DST-FIST program for infrastructural facilities is acknowledged.

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  1. Plant Biotechnology Laboratory, Department of Botany, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata, 700118, India

    Sk Moquammel Haque & Biswajit Ghosh

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  1. Sk Moquammel Haque
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  2. Biswajit Ghosh
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Correspondence to Biswajit Ghosh.

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Haque, S.M., Ghosh, B. Somatic Embryogenesis and Synthetic Seed Production—a Biotechnological Approach for True-to-Type Propagation and In Vitro Conservation of an Ornamental Bulbaceous Plant Drimiopsis kirkii Baker.. Appl Biochem Biotechnol 172, 4013–4024 (2014). https://doi.org/10.1007/s12010-014-0817-2

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  • DOI: https://doi.org/10.1007/s12010-014-0817-2

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