Quantitative Evaluation of Chickpea Fusarium Wilt

Fusarium Oxysporum f. sp. Ciceris (FOC) is the causal agent of Fusarium wilt, a destructive and widespread disease of chickpea. Rapid and accurate identification and detection of plant pathogens are essential for timely Disease Management (DM) strategies with appropriate measures. This study aimed to quantitatively determine FOC by using Quantitative Real-Time Polymerase Chain Reaction (qPCR) technique with specific primer pairs [Histone (H۳) and Ribosomal (J۵)] in seed, root, and root collar, and to discriminate it from other pathogenic fungi [Fusarium Oxysporum formae speciales (FO f. sp.) and Ascocyhta rabiei]. Total RNAs were isolated, converted to cDNAs (limit of ۵ ng/rxn.-۰.۰۵ pg/rxn.) and used as template for qPCR studies. The FOC was detected in plant samples starting from the first day after inoculation. The FOC was detected in root, root collar and seed samples and was differentiated by qPCR assay from other pathogenic fungi. Melting curves, in which no primer dimers and non-specific complementation were observed, presented a single peak. Quantification was successfully performed using specific H۳ and J۵ primer pairs (P< ۰.۰۵), and the FOC was distinguished from other pathogenic fungi with J۵ primer (P< ۰.۰۵). The results of these studies may support the development of new biochemical and molecular methods that allow direct, faster and more accurate determination of pathogens. Thus, it will also enable us to reduce the losses caused by diseases and the costs of DM.

کلیدواژه ها:

Fusarium oxysporum f. sp. ciceris ، Fusarium oxysporum formae speciales ، qPCR ، RT-qPCR ، Wilting and Yellowing.