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<p><i>Aegle marmelos</i> is one of the most highly demanded medicinal plants used in the indigenous system of medicine in South Asian countries. Exploitation by the industries, indiscriminate or unregulated harvesting practices with no concern for the sustainability of the resource and lack of organized cultivation, has enlisted the plant as a near threatened species by the International Union for Conservation of Nature. The study envisaged the conservation of <i>A. marmelos</i> synseeds using single node segments from in vitro maintained plants via encapsulation-dehydration method of cryopreservation. The preconditioning, encapsulation, preculture and dehydration steps of cryopreservation were standardized in the study. Sucrose 0.1&#xa0;M in semi solid MS (Murashige and Skoog) medium for 7days was selected as the best preconditioning treatment, which produced maximum shoot proliferation (5.50 shoots per culture) when cultured on basal medium (MS + Benzyl adenine (BA) 2 mg L<sup>− 1</sup> + Indole-3-butyric acid (IBA) 0.5 mg L<sup>− 1</sup>). Synseeds were prepared by encapsulating the buds in calcium alginate matrix (formed by sodium alginate 3.5% with Calcium chloride 100 mM), when precultured to liquid MS medium supplemented with sucrose 0.5&#xa0;M and dimethyl sulfoxide (DMSO) 3% for 3 days, gave maximum shoot proliferation (3.67 shoots per explant) in the modified basal medium (½ MS (with half the concentration of macronutrients) + BA 2 mg L<sup>− 1</sup> + IBA 0.5 mg L<sup>− 1</sup>) The desiccated synseeds were then cryopreserved in liquid nitrogen for 1 week, followed by thawing for 30–60&#xa0;s at 40&#xa0;°C and inoculated on to recovery medium ½ MS + BA 2 mg L<sup>− 1</sup> + IBA 0.5 mg L<sup>− 1</sup>. At 6&#xa0;h of desiccation, and moisture content of 19.50%, maximum survival (66.67%) and regeneration (50%) was obtained. The genetic stability of cryo-regenerated plantlets was confirmed using Inter Simple Sequence Repeat (ISSR) markers. This is a pioneer report on cryopreservation protocol that could be utilized for long term conservation of <i>A. marmelos.</i></p>

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Cryopreservation of bael (Aegle marmelos L.Corr.) using encapsulated axillary buds

  • 期刊论文

  • 发表日期：

    2025年10月31日

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  • E. Deepa,
  • Deepa S. Nair,
  • Swapna Alex,
  • K. B. Soni,
  • M. M. Viji,
  • B. Parvathi

摘要

Aegle marmelos is one of the most highly demanded medicinal plants used in the indigenous system of medicine in South Asian countries. Exploitation by the industries, indiscriminate or unregulated harvesting practices with no concern for the sustainability of the resource and lack of organized cultivation, has enlisted the plant as a near threatened species by the International Union for Conservation of Nature. The study envisaged the conservation of A. marmelos synseeds using single node segments from in vitro maintained plants via encapsulation-dehydration method of cryopreservation. The preconditioning, encapsulation, preculture and dehydration steps of cryopreservation were standardized in the study. Sucrose 0.1 M in semi solid MS (Murashige and Skoog) medium for 7days was selected as the best preconditioning treatment, which produced maximum shoot proliferation (5.50 shoots per culture) when cultured on basal medium (MS + Benzyl adenine (BA) 2 mg L− 1 + Indole-3-butyric acid (IBA) 0.5 mg L− 1). Synseeds were prepared by encapsulating the buds in calcium alginate matrix (formed by sodium alginate 3.5% with Calcium chloride 100 mM), when precultured to liquid MS medium supplemented with sucrose 0.5 M and dimethyl sulfoxide (DMSO) 3% for 3 days, gave maximum shoot proliferation (3.67 shoots per explant) in the modified basal medium (½ MS (with half the concentration of macronutrients) + BA 2 mg L− 1 + IBA 0.5 mg L− 1) The desiccated synseeds were then cryopreserved in liquid nitrogen for 1 week, followed by thawing for 30–60 s at 40 °C and inoculated on to recovery medium ½ MS + BA 2 mg L− 1 + IBA 0.5 mg L− 1. At 6 h of desiccation, and moisture content of 19.50%, maximum survival (66.67%) and regeneration (50%) was obtained. The genetic stability of cryo-regenerated plantlets was confirmed using Inter Simple Sequence Repeat (ISSR) markers. This is a pioneer report on cryopreservation protocol that could be utilized for long term conservation of A. marmelos.