Cumin Transformation for Resistance to Fungal Diseases

Cumin (Cuminum cyminum L.) is an economically valuable medicinal and spice plant that is highly susceptible to fungal pathogens, including Fusarium oxysporum f. sp. cumini, which can cause yield losses of up to ۸۰%. In this study, a multi-gene construct (pBI۱۲۱) carrying chitinase, β-۱,۳-glucanase, and PRP۱ genes, each driven by an independent CaMV ۳۵S promoter and accompanied by a nptII selectable marker, was introduced into cumin through Agrobacterium tumefaciens–mediated transformation. Cotyledon explants were co-cultivated with Agrobacterium strain EHA۱۰۵ at OD۶۰۰ = ۰.۵, followed by selection on MS medium containing ۵۰ mg L-۱ kanamycin. A total of nine transgenic lines were confirmed by PCR amplification of specific bands at approximately ۸۷۰ bp (glucanase), ۶۸۰ bp (chitinase), and ۵۸۰ bp (PRP۱), corresponding to a transformation efficiency of about ۱.۵%. Protein extracts from transgenic plants exhibited clear antifungal activity against Fusarium oxysporum in vitro, with inhibition zones averaging ۶.۳ ± ۰.۵ mm and ۹.۸ ± ۰.۷ mm for ۵۰ µg and ۱۰۰ µg protein concentrations, respectively. No inhibition was observed in extracts from non-transgenic plants or buffer controls. The antifungal effect is attributed to synergistic action among chitinase, glucanase, and PRP۱ proteins, which degrade fungal cell wall components and activate defense signaling pathways. These results demonstrate the feasibility of Agrobacterium-mediated transformation in cumin and highlight the potential of multi-gene stacking strategies for enhancing fungal resistance in medicinal plants. This work represents an important step toward developing disease-resistant cumin genotypes and reducing reliance on chemical fungicides in sustainable agriculture.

کلیدواژه ها:

Agrobacterium ، antifungal bioassay ، chitinase ، Cuminum cyminum ، Fusarium oxysporum ، glucanase ، PRP۱