Ferdous-E-Elahi, Md. Mahamudul Hasan, Monirul Islam, Sayma Tahsin Nira, Tasfia Hassan Tabassum, Md. Abu Raihan and Md. Monirul Islam
Abstract
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Cabbage (Brassica oleracea var. capitata) is one of the major winter vegetables grown in Bangladesh, recognized for its dietary value and economic importance. During November 2023, cabbage leaves exhibiting typical leaf spot symptoms were collected from the Bangladesh Agricultural Research Institute (BARI), Gazipur, to identify the associated pathogen. The infected leaves showed dark brown to black necrotic lesions with concentric rings characteristic of Alternaria leaf spot. The pathogen was isolated on Potato Dextrose Agar (PDA) and purified using a single-conidium isolation technique. Morphological characterization revealed colony and conidial features consistent with Alternaria alternata. To confirm the pathogen identity, molecular characterization was performed using the internal transcribed spacer (ITS) region of rDNA. Amplification with ITS1/ITS4 primers generated a 510 bp fragment, and the resulting sequence was deposited in GenBank under accession number OP585635. Comparison of the obtained sequence against the GenBank database revealed a high degree of homology (99%) with previously deposited A. alternata sequences. The phylogenetic tree also placed the isolate in the same evolutionary group as A. alternata, with 88% bootstrap support. Species-specific PCR using A. alternata-specific primers produced the expected 184 bp amplicon, providing additional confirmation of species identity. Pathogenicity was evaluated using a detached-leaf assay. Leaves inoculated with the fungal isolate exhibited characteristic disease symptoms comparable to those recorded in naturally infected plants, while no symptoms were detected on the control leaves. Following symptom development, the fungus was recovered from infected tissues and its identity was verified using an A. alternata-specific PCR assay, satisfying the requirements of Koch’s postulates. The combined evidence obtained from cultural and microscopic observations, ITS-rDNA sequencing, phylogenetic inference, species-specific molecular detection, and pathogenicity assessment demonstrated that A. alternata was responsible for the leaf spot symptoms observed on cabbage. To our knowledge, this study constitutes the first molecularly confirmed record of A. alternata infecting cabbage in Bangladesh. The information generated will support future disease monitoring efforts and contribute to the formulation of appropriate management practices for sustainable cabbage production.