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Direct Shoot From Root and True-to-Type Micropropagation of Limonium “Misty Blue” in Partially Immersed Culture on an Aluminum Mesh Raft

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Micropropagation Methods in Temporary Immersion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2759))

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Abstract

Commercial plant tissue culture now primarily serves the ornamental horticulture industry. The main pillars of the commercial tissue culture business are scalability of production, cost reduction, limited labor involvement, high quality, and genetic homogeneity of propagated plants. Based on these requirements, the current protocol employs a partially immersed liquid culture medium supported by a flexible aluminum mesh raft with a wire stand to facilitate shoot organogenesis from the horizontally placed root explants and hold the plants upright for shoot multiplication and rooting of Limonium Misty Blue. It is a florist crop that is in high demand as both dried and fresh flower fillers in various floral decorations. The majority of cultivated Limonium or statice cultivars are heterozygous in nature and propagate commercially through in vitro propagation to cater to the huge demand for planting materials needed for flower production. This is the first protocol to describe direct shoot organogenesis from the roots in a liquid half-component of Murashige and Skoog’s (1962) (MS) basal medium supplemented with 1.6 μM NAA and 1.1 μM BA. The regenerated shoots are multiplied and rooted at the same time on the raft in a MS-based liquid culture medium that included 0.44 μM BA and 1.07 μM NAA. In comparison to agar-gelled medium, plants cultured in liquid medium grow more quickly without any signs of hyperhydricity. In liquid medium, a clump of 4–5 shoots is formed from a single shoot explant within 4 weeks and are rooted simultaneously within 6 weeks. On average, seven explants may fit on each raft, so on average, 25 healthy plants are produced from a single bottle. The regenerated plants are easily hardened in the greenhouse, and using ISSR-based molecular markers, the genetic homogeneity of the randomly selected hardened plants can be determined.

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Acknowledgments

This work was financially supported by WBHESTB, Govt. of West Bengal [846 (Sanc.)/ST/P/S and T/2G-11/2018; June 19, 2019]; DST-PURSE PROGRAMME, GOI; Savitribai Jyotirao Phule Fellowship for Single Girl Child (UGCES-22-GE-WES-F-SJSGC-16026, UGCES-22-GE-WES-F-SJSGC-16035), RUSA 10 Component (CH&E) [IP/RUSA(C-10)/17/2021 dated 26.11.2021]; and Personal Research Grants (PRG) for teachers and URS provided by the University of Kalyani, West Bengal, India. The assistance of Purbasa Kole during the experiment is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Molecular Biology and Biotechnology, Faculty of Science, University of Kalyani, Kalyani, West Bengal, India

    Priyanka Raha, Gourab Saha, Ishita Khatua & Tapas Kumar Bandyopadhyay

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  1. Priyanka Raha
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  2. Gourab Saha
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  3. Ishita Khatua
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  4. Tapas Kumar Bandyopadhyay
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Correspondence to Tapas Kumar Bandyopadhyay .

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Editors and Affiliations

  1. Unidad Guerrero Negro, Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), Guerrero Negro, Baja California Sur, Mexico

    Marco A. Ramírez-Mosqueda

  2. Facultad de Ciencias Químicas, Universidad Veracruzana, Orizaba, Veracruz, Mexico

    Carlos A. Cruz-Cruz

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Raha, P., Saha, G., Khatua, I., Bandyopadhyay, T.K. (2024). Direct Shoot From Root and True-to-Type Micropropagation of Limonium “Misty Blue” in Partially Immersed Culture on an Aluminum Mesh Raft. In: Ramírez-Mosqueda, M.A., Cruz-Cruz, C.A. (eds) Micropropagation Methods in Temporary Immersion Systems. Methods in Molecular Biology, vol 2759. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3654-1_11

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  • DOI: https://doi.org/10.1007/978-1-0716-3654-1_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3653-4

  • Online ISBN: 978-1-0716-3654-1

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