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In vitro propagation and ultrastructural studies of somatic embryogenesis of Ledebouria ovatifolia

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Abstract

Simple and efficient in vitro plant production systems were established for Ledebouria ovatifolia (Bak.) Jess. Adventitious shoots were best produced from leaf explants with Murashige and Skoog medium containing 5 μM thidiazuron and 2 μM naphthaleneacetic acid, and from organogenic callus with Murashige and Skoog medium containing 2 μM indole-3-acetic acid, 5 μM thidiazuron, and 30 μM glutamine. Indole-3-butyric acid and 25 μM phloroglucinol were effective for rooting of shoots. Embryogenic callus was induced on semi-solid medium containing growth regulators. The highest numbers of somatic embryos, 43.2–35.6 (globular to cotyledonary stages, respectively) from friable, embryogenic callus were obtained on liquid medium with 15 g L−1 sucrose, 10 μM glutamine, 0.1 μM picloram, and 0.2 μM thidiazuron. Seventy-three percent of somatic embryos germinated on semi-solid medium with 15 g L−1 sucrose, 0.3 μM gibberellic acid, 0.3 μM phloroglucinol, and Murashige and Skoog macronutrients. All plantlets were successfully acclimatized in the greenhouse. Production of clonal plants was confirmed by features of embryoids using light and transmission electron microscopy, which detected cytoplasmic components including many mitochondria, lipid bodies together with starch grains, chloroplasts, Golgi apparatuses, vacuoles, and nuclei. The reported developmental system reinforced the importance of nutritional and hormonal effects as well as the effect of phloroglucinol on in vitro plant production. Histological and ultrastructural studies demonstrated the bipolar structure, and viability of somatic embryoids. The micropropagation and somatic embryogenesis protocols reported here provide systems for germplasm conservation and large-scale clonal propagation, and for pharmacological, and genetic transformation studies.

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Acknowledgments

Financial support by the National Research Foundation (NRF), Pretoria and the University of KwaZulu-Natal, Pietermaritzburg is gratefully acknowledged. The authors are grateful to Subashen Naidu and all other Staff of the Microscopy & Microanalysis Unit (MMU), UKZN, Pietermaritzburg for microscopic assistance.

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Authors and Affiliations

  1. Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal Pietermaritzburg, Private Bag X01, Scottsville, 3209, South Africa

    Ponnusamy Baskaran, Aloka Kumari, Devashan Naidoo & Johannes Van Staden

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  1. Ponnusamy Baskaran
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  2. Aloka Kumari
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  3. Devashan Naidoo
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  4. Johannes Van Staden
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Correspondence to Johannes Van Staden.

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Editor: Jorge Canhoto

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Baskaran, P., Kumari, A., Naidoo, D. et al. In vitro propagation and ultrastructural studies of somatic embryogenesis of Ledebouria ovatifolia . In Vitro Cell.Dev.Biol.-Plant 52, 283–292 (2016). https://doi.org/10.1007/s11627-016-9762-9

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  • DOI: https://doi.org/10.1007/s11627-016-9762-9

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