Optimizing callus induction and indirect organogenesis in non-dormant corm explants of Gloriosa superba (L.) via media priming

Dexter Achu Mosoh, Ashok Kumar Khandel, Sandeep Kumar Verma, W. Vendrame
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Abstract

Gloriosa superba L. is an endangered ornamental plant of significant medicinal, commercial, and cultural value. This study establishes an efficient protocol for rapid in vitro propagation of this plant species through callus-mediated organogenesis using non-dormant corm explants.Plant tissue culture techniques were employed to facilitate the in vitro regeneration process of Gloriosa superba L. This encompassed various stages, including the acquisition of plant material, surface sterilization, formulation of growth media, and the execution of callogenesis, shooting, rooting, and acclimatization experiments.Investigation into the impact of plant growth regulators on callogenesis unveiled diverse callus morphologies, dependent on regulator type and concentration, with the NAA and KN combination emerging as the most effective for callus induction. Specifically, the callus induction medium supplemented with 1.5 mg L−1 NAA, 0.5 mg L−1 KN, and 10 mg L−1 casein hydrolysate (CH) achieved remarkable results, boasting an 81.25% callus induction rate and a substantial callus biomass fresh weight of 333 mg. Furthermore, the combination of BAP and NAA facilitated optimal shoot primordia induction and shooting in callus derived from non-dormant corm explants. The shoot induction medium, enriched with 2.0 mg L−1 BAP, 0.5 mg L−1 NAA, 5 mg L−1 CH, and 20% (v/v) coconut water (CW), resulted in an impressive 83.33% shoot primordia formation rate and an average of 6.86 shoot primordia per callus. For root induction, the medium supplemented with 1.0 mg L−1 IBA displayed a high root induction rate of 81.25% and robust rooting (+++). Successful acclimatization of in vitro plantlets in controlled environments and shade net houses culminated in thriving growth, and upon transplantation into a garden soil blend (garden soil : sand : vermiculite, 2:1:1, v/v), with direct sunlight exposure, the plantlets demonstrated commendable development and form, boasting a 69% survival rate at the seven-week mark. These findings offer a robust foundation for scalable and sustainable propagation strategies, ensuring the conservation and utilization of this valuable species.
通过培养基引物优化非休眠球茎外植体的胼胝体诱导和间接器官发生
Gloriosa superba L.是一种濒危观赏植物,具有重要的药用、商业和文化价值。本研究利用非休眠期球茎外植体,通过胼胝质介导的器官发生,建立了快速体外繁殖该植物物种的有效方案。对植物生长调节剂对胼胝发生影响的研究揭示了胼胝形态的多样性,这取决于调节剂的类型和浓度,其中 NAA 和 KN 的组合对胼胝诱导最有效。具体而言,添加了 1.5 mg L-1 NAA、0.5 mg L-1 KN 和 10 mg L-1 酪蛋白水解物(CH)的胼胝体诱导培养基取得了显著效果,胼胝体诱导率达 81.25%,胼胝体生物量鲜重达 333 mg。此外,BAP 和 NAA 的组合还能促进非休眠期球茎外植体产生的胼胝体获得最佳的芽原基诱导和生长。在芽诱导培养基中添加 2.0 mg L-1 BAP、0.5 mg L-1 NAA、5 mg L-1 CH 和 20%(v/v)椰子水(CW)后,芽原基形成率达到 83.33%,每个胼胝体平均形成 6.86 个芽原基。在根诱导方面,添加 1.0 mg L-1 IBA 的培养基显示出 81.25% 的高根诱导率和强大的生根能力(+++)。离体小植株在受控环境和遮阳网房中成功适应环境后,生长茁壮,移栽到混合园土(园土:沙:蛭石,2:1:1,v/v)中并直接接受阳光照射后,小植株的发育和形态值得称赞,七周存活率高达 69%。这些发现为可扩展和可持续的繁殖策略奠定了坚实的基础,确保了这一珍贵物种的保护和利用。
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