فهرست نشریات علمی دسترسی آزاد ایران

Canola (Brassica napus L.), a globally significant source of edible vegetable oil, is highly susceptible to various phytopathogenic fungi. One strategy to bolster its resistance involves genetic engineering through the introduction of genes encoding antifungal peptides, which are plant-derived defense proteins. In this study, the Dabb1 gene was isolated from Arabidopsis thaliana via polymerase chain reaction (PCR) using specific primers. The amplified fragment was subsequently confirmed through PCR analysis, restriction digestion, and sequencing. This gene was then overexpressed in the R-line Hyola 308 of B. napus. Transformation of cotyledonary petioles was successfully accomplished using the pBIDAB construct, which carries the Dabb1 gene under the control of the CaMV35S promoter, via Agrobacterium-mediated transformation. Integration of the transgene in the resulting transformants was confirmed by PCR. A total of twenty-one PCR-positive plants were obtained from 450 explants, yielding a transformation efficiency of 4.66%. Antifungal assays were conducted against Sclerotinia sclerotiorum, Fusarium oxysporum, Alternaria solani, Botrytis cinerea, and Verticillium dahliae. These assays utilized total protein extracted from transgenic plants and assessed through spore germination inhibition and disk diffusion methods. The extracted proteins from transgenic plants demonstrated the ability to reduce spore germination and inhibit fungal growth. Furthermore, a biosafety assessment using a hemolysis test indicated that the protein extracted from transgenic canola exhibits no toxic effects on mammalian cells.