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Design, Engineering, and Expression of the Gene Encoding FeAP Antifungal Peptide in Tobacco with Aimed of Developing a Novel Strategy for Controlling the Ascochyta rabiei Fungus
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Sahba Toosi   , Saeid Malekzadeh Shafaroudi * , Farhad Shokouhifar , Mojtaba Mamarabadi , Nasrin Moshtaghi |
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Abstract: (79 Views) |
Antifungal peptides are among the effective strategies for combating pathogenic fungi such as Ascochyta rabiei. This study was conducted to investigate the antifungal activity of the FeAP peptide against A. rabiei. Therefore, an expression construct was designed and molecular dynamics analyses were performed to evaluate the stability of the antifungal peptide. The target gene was synthesized and inserted into the pCAMBIA3301expression vector. The construct was transformed into Agrobacterium and then tobacco plants were infected with this bacterium. Successful transformation of tobacco plants was confirmed using the GUS reporter gene and RT-PCR analysis. The FeAP peptide purified using a Ni-NTA affinity column, and its antifungal activity was evaluated using the well diffusion method against A. rabiei. The results of the molecular dynamics analyses indicated that the designed construct enabled the production of a stable peptide. Production of the antifungal peptide, with an approximate molecular weight of 11 kDa, was confirmed by SDS-PAGE. A clear zone with a diameter of 14 mm demonstrated inhibition of A. rabiei growth at a concentration of 320 mg/ml. Furthermore, the findings of this study may contribute to the development of chickpea plants resistant to A. rabiei.
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| Keywords: Agrobacterium, Gene delivery, Antifungal peptide, Molecular dynamics |
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Full-Text [PDF 1081 kb]
(70 Downloads)
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Type of Study: Research |
Subject:
Special
Received: 2025/12/1 | Accepted: 2026/02/17 | Published: 2026/05/20
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