Callus induction, frond regeneration, and genetic transformation of the aquatic plant Lemna minor using Agrobacterium tumefaciens

Document Type : Research paper

Authors

National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.

Abstract

Molecular farming involves the large-scale cultivation of plants to produce recombinant proteins for diverse biotechnological applications, including biopharmaceuticals, veterinary products, research tools, and dietary supplements. Members of the Lemnaceae family, small aquatic plants typically found in swamps and lakes, have garnered attention for molecular farming due to their rapid growth, vegetative propagation, and high protein content. In this study, the Iranian ecotype of Lemna minor plants was collected from Guilan Province in northern Iran. To induce callus formation, five different media—Murashige and Skoog (MS), 1/2MS, B5, Schenk and Hildebrandt (SH), and Hoagland —were evaluated with various combinations of hormones and sucrose concentrations. The impact of acetosyringone in the co-cultivation medium on transformation efficiency was also examined. The findings revealed that Hoagland and standard MS basal media, combined with 12 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), 4.8 mg/L 6-Benzylaminopurine (BAP), and 2% sucrose, significantly enhanced callus induction. For frond regeneration, Hoagland medium supplemented with 2% sucrose, 1.1 mg/L kinetin, 4.5 mg/L indole-3-acetic acid (IAA), and 1 mg/L thidiazuron (TDZ), 4 mg/L IAA showed the highest performance. Additionally, the optimal transformation efficiency was achieved using 200 µM acetosyringone. This study provides an efficient and reproducible protocol for callus induction and frond regeneration in L. minor, offering valuable insights for molecular farming and recombinant protein production.

Keywords

References

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History

  • Receive Date: 06 March 2025
  • Revise Date: 21 September 2025
  • Accept Date: 22 September 2025

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