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<p>Legume species play a crucial role in human nutrition but are recalcitrant to various <i>in vitro</i> morphogenic processes. A repeatable technique was developed to enhance the frequency of direct shoot regeneration from intact seedlings of moth bean (<i>Vigna aconitifolia</i> (Jacq.) Marechal) and lentil (<i>Lens culinaris</i> Medik.) using thidiazuron (TDZ) flooding by eliminating the need for intermediate callus stages. In moth bean and lentil seedlings, shoots were induced from the cotyledonary node when cultured on 1.0&#xa0;µM TDZ. Remarkably, this effect was significantly enhanced with shoots emerging from the epicotyl when the seedlings were flooded with the same concentration of TDZ. A comparative analysis revealed that TDZ outperformed BAP (benzylaminopurine), as 90% of seedlings exhibited shoot emergence on 1.0&#xa0;µM TDZ, compared to only 68 to 70% on 10.0&#xa0;µM BAP. Seedlings grown on TDZ or BAP showed substantial inhibition of root growth and shoot apex expansion, particularly in flooded cultures. The observed shoot apex expansion followed by direct shoot formation on the epicotyl represented a significant advancement in this area. This study emphasized the importance of reliable shoot regeneration techniques as a foundation for whole plant regeneration and genetic transformation in legumes by avoiding the callus stages.</p>

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Flood-induced in vitro shoot regeneration in Vigna aconitifolia and Lens culinaris using thidiazuron: a novel stress-driven approach

  • 期刊论文

  • 发表日期：

    2025年11月3日

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  • Renu Soni,
  • Gayatri Tripathi

摘要

Legume species play a crucial role in human nutrition but are recalcitrant to various in vitro morphogenic processes. A repeatable technique was developed to enhance the frequency of direct shoot regeneration from intact seedlings of moth bean (Vigna aconitifolia (Jacq.) Marechal) and lentil (Lens culinaris Medik.) using thidiazuron (TDZ) flooding by eliminating the need for intermediate callus stages. In moth bean and lentil seedlings, shoots were induced from the cotyledonary node when cultured on 1.0 µM TDZ. Remarkably, this effect was significantly enhanced with shoots emerging from the epicotyl when the seedlings were flooded with the same concentration of TDZ. A comparative analysis revealed that TDZ outperformed BAP (benzylaminopurine), as 90% of seedlings exhibited shoot emergence on 1.0 µM TDZ, compared to only 68 to 70% on 10.0 µM BAP. Seedlings grown on TDZ or BAP showed substantial inhibition of root growth and shoot apex expansion, particularly in flooded cultures. The observed shoot apex expansion followed by direct shoot formation on the epicotyl represented a significant advancement in this area. This study emphasized the importance of reliable shoot regeneration techniques as a foundation for whole plant regeneration and genetic transformation in legumes by avoiding the callus stages.