Identification and biological control of bacterial leaf spot disease of cucurbits

M. E. Khan Chowdhury, Md. Enamul Haque, Mehrun Nesa, Biswanath Sikdar, Md. Faruk Hasan
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Abstract

Objective: The present study was conducted for isolation and detection of the phyto-pathogen responsible for bacterial leaf spot disease of cucurbits as well as evaluation of its biological control techniques. Methods: The pathogen of the disease was isolated from infected leaf of bitter gourd and cultured on Luria-Bertani (LB) growth medium. The morphological tests and biochemical tests revealed the isolated bacteria as gram negative. Furthermore, advanced molecular technique was applied to facilitate proper detection of the isolated bacteria. The PCR of the bacterial genomic DNA using specific primers generated clear band of approximately 1450 bp in gel electrophoresis. Results: In gram staining test, the bacterial strain was found to be rod shaped and pinkish in color and Gram-negative. All the biochemical test showed positive without mannitol salt agar. The nucleotide sequencing of 16S rDNA gene of the bacterial isolate showed 86.02% similarities with the original sequence of Xanthomonas cucurbitae. In antibiotic sensitivity assay, the antibiotic cefixime showed highest 28.0±0.7 mm diameter of inhibition zone against the tested bacteria. The highest antibacterial activity was 8.0±0.5 mm zone of inhibition by of Allium sativum. Conclusion: The present findings may be benevolent for the detection, characterization and development of biological control techniques to prevent the leaf spot disease of cucurbits crops in Bangladesh. It may help to find out the specific signaling pathway techniques for plant microbes relationship for disease formation in future.
葫芦细菌性叶斑病的鉴定和生物防治
研究目的本研究旨在分离和检测导致葫芦细菌性叶斑病的植物病原体,并评估其生物防治技术。研究方法从受感染的苦瓜叶片中分离出病原菌,并在 Luria-Bertani (LB) 生长培养基上进行培养。形态学检测和生化检测显示,分离出的细菌为革兰氏阴性菌。此外,还采用了先进的分子技术,以便对分离出的细菌进行正确检测。使用特定引物对细菌基因组 DNA 进行 PCR,在凝胶电泳中产生了约 1450 bp 的清晰条带。检测结果在革兰氏染色试验中,发现该细菌菌株呈棒状、粉红色、革兰氏阴性。所有生化测试均显示阳性,无甘露醇盐琼脂。细菌分离物 16S rDNA 基因的核苷酸测序结果显示,其与葫芦黄单胞菌原始序列的相似度为 86.02%。在抗生素敏感性试验中,抗生素头孢克肟对受试细菌的抑菌区直径最大,为 28.0±0.7 mm。薤白的抑菌区为 8.0±0.5 毫米,抗菌活性最高。结论本研究结果可能有助于检测、鉴定和开发生物防治技术,以预防孟加拉国葫芦科作物叶斑病。它可能有助于找出植物微生物与疾病形成关系的特定信号通路技术。
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