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Droplet vitrification: a lifeline for long-term conservation of threatened species Garcinia indica

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Abstract

Cryopreservation is a promising technique for the ex situ long-term conservation of plant biodiversity particularly of species that are not amenable to seed bank conservation. Garcinia indica (Thouars) Choisy is a species endemic to the biodiversity hotspot of the Western Ghats of India. Conservation of this species is difficult as it produces recalcitrant seeds, and therefore, it can only be conserved in the field genebanks or in their natural habitats. This high-value fruit tree species is listed as vulnerable due to the rapid loss of the natural population, and therefore, the conservation of its genetic diversity is imperative. This study described the first successful cryopreservation protocol for G. indica through a modified droplet vitrification technique using in vitro–derived shoots conserved in the In Vitro Gene Bank of the Indian Council of Agricultural Research - National Bureau of Plant Genetic Resources (ICAR-NBPGR), New Delhi. Among vitrification (V) and droplet vitrification (DV) techniques, only the DV technique yielded explant regeneration after cryopreservation. Apical shoots (2 mm long) collected from 24-wk-old explants (IC638183), without any pre-culture, showed maximum regeneration (51.76%) on Murashige and Skoog (MS) medium supplemented with 2.22 μM 6-benzylaminopurine (BAP). The regenerated shoots were hardened in a mist chamber successfully (85% survival), which is essential for the future restoration of the species in the field. With genotypic-dependent variation, this protocol was applicable to the other three accessions (IC638184, IC638185, and IC638186) with an average of 43.7% regeneration after cryopreservation, which can be implemented for the effective conservation of G. indica germplasm.

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Acknowledgements

The authors greatly acknowledge the Director, ICAR-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, for supporting this study. Thanks to Dr J.K. Ranjan, Principal Scientist, Indian Agricultural Research Institute (IARI), New Delhi, for helping with the statistical analysis. The authors are thankful to Dr K. Pradheep, Principal Scientist and Officer-in-Charge, ICAR-NBPGR, Regional Station-Thrissur, for his valuable input. The first author thanks the Belgian Government for training on “Cryopreservation of tropical crop species” at the Laboratory of Tropical Crop Improvement, KU Leuven, Belgium.

Funding

This work was done as part of the intra-mural project “In vitro conservation of tropical fruit crops” (project code: PGR/TCCUBUR-DEL-01.05) at ICAR-NBPGR that was funded by the Indian Council of Agricultural Research (ICAR).

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Authors and Affiliations

  1. Division of Germplasm Conservation, Indian Council of Agricultural Research (ICAR)-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, 110012, India

    Vartika Srivastava

  2. Alliance of Bioversity International and CIAT, KU Leuven, 3001, Leuven, Belgium

    Bart Panis

  3. National Agricultural Higher Education Project (NAHEP), ICAR, Krishi Anusandhan Bhawan-II, Pusa Campus, New Delhi, 110012, India

    Anuradha Agrawal

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  1. Vartika Srivastava
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VS: conceptualization, methodology, formal analysis, investigation, writing—original draft. AA: writing—review and editing, supervision. BP: writing—review and editing, supervision

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Correspondence to Anuradha Agrawal.

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Srivastava, V., Panis, B. & Agrawal, A. Droplet vitrification: a lifeline for long-term conservation of threatened species Garcinia indica. In Vitro Cell.Dev.Biol.-Plant (2024). https://doi.org/10.1007/s11627-024-10423-2

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