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PCR and Sequencing Base Detection of Gummosis Disease on Citrus aurantifolia Caused by Lasiodiplodia theobromae and Evaluation of Its Antagonisms

Md. Faruk Hasan, Md. Asadul Islam, Biswanath Sikdar

Journal of Advances in Microbiology · pp. 77–90 · Published 2 Apr 2020

10.9734/jamb/2020/v20i330230

Abstract

Diplodia gummosis disease caused by Lasiodiplodia theobromae is an economically important postharvest fruit decay that occurs on all types of citrus grown in Bangladesh. The present investigation was conducted to isolate and identify the pathogenic fungus responsible for postharvest diplodia gummosis disease (DGD) of citrus as well as the evaluation of its biological control through microbial antagonists. DGD causing pathogen was identified by physiological, morphological and molecular methods. The pathogenic fungus was isolated from the surface of postharvest lime fruits. The optimum growth of the fungi was observed in maltose, 2% NaCl, 15% sugar and pH 7 containing PDA medium at 35⁰C. PCR products of the internal transcribed spacer (ITS) region of the fungus showed approximately 650 bp size clear band in gel electrophoresis. The amplified region of the fungus showed 99.62% similarities with the sequences of L. theobromae. Artificially inoculation of the fungus in malta, musumbi, sweet orange, lime, and guava fruits showed similar size clear band and typical diplodia gummosis disease symptom. Methanol extracts of Datura metal displayed the highest inhibition (75.25%) against the isolated fungus. Non-pathogenic fungi Trichoderma viride showed the highest antagonistic efficiency followed by Neofusicoccum mangifera against the isolated fungus. The tested soil bacteria did not show significant antagonistic activity against the isolated fungus. Therefore, the DGD of citrus control system should be integrated into the overall citrus postharvest decay control system to reduce all citrus postharvest diseases and to protect fresh citrus fruit values.

Citrus aurantifolia PCR sequencing Lasiodiplodia theobromae; antagonisms.

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