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This study aimed to isolate and characterise the pathogen associated with Alternaria leaf spot on broccoli and to evaluate the inhibitory effects of fungicides against it. We isolated and identified the fungal pathogen as Alternaria sp. using morphological and cultural methods. Based on the aligned sequences of the internal transcribed spacer (ITS) and molecular phylogenetic analysis by the neighbour-joining method, the isolates (Ab1 and Ab2) were confirmed as Alternaria alternata. The conidia of the isolates were dark brown, cylindrical, obclavate to muriform. The conidiophores were olivaceous brown, septate, and branched. The conidial morphology of the isolates ranged from 52.4–92.4 × 10–20 μm with 2–6 transverse and 0–3 longitudinal septa. Both isolates yielded positive results in the pathogenicity test on broccoli leaves by developing brown and circular spots with concentric rings on the leaves surrounded by yellow halos. The culture studies revealed that the maximum growth of the pathogen was obtained at 30 °C and pH 6.0. Tilt 250 WC showed the highest potential in suppressing the mycelial growth of the A. alternata in vitro at a concentration as low as 50 µg/mL. The results from this study contributed to the positive identification of the pathogen and characterised A. alternata as a destructive pathogen of broccoli which may be successfully controlled by the fungicide Tilt.

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