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Original Research Article

Characterization and Antagonistic Control Measures of Pichia kudriavzevii Yeast like Fungus in Brown Rot Disease of Citrus sinensis (L.) Osbeck

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

Professor, Joarder DNA and Chromosome Research Lab., Department of Genetic Engineering and Biotechnology, Faculty of Life and Earth Sciences, University of Rajshahi, Rajshahi-6205,Bangladesh.

Ind.J.Applied.Microbiol. 2019 .22(1) : 31-48

*Corresponding author

Abstract:

Citrus sinensis is one of the most important Vitamin C containing fruit all over the  world which is badly infected by brown rot disease (BRD). The present research work was  designed to isolate and characterize the pathogenic fungus responsible for BRD of C. sinensis as  well as evaluation of its antagonistic control measures. The causal agent of BRD was isolated and  characterized using physiological, morphological and molecular methods. The pathogenic fungus  was isolated from the skin of sweet orange. The isolated fungus was grown in different nutrients  media, temperature, pH, sugar concentrations and different carbohydrates. The highest growth of  the fungi was observed in 5% fructose containing PD broth media at 370C, pH 7.5 and 0.05gm/L  NaCl concentration. PCR amplification of the isolated fungal DNA by internal transcribed spacer  (ITS) region products showed approximately 650bp size of clear cut band. sequencing analysis  showed that the amplified its region of the fungus (GenBank Submission# SUB5672073) has  99.80% similarities with the sequences of Pichia kudriavzevii isolate 11-577 its-1 partial  sequence. In pathogenicity assay, artificially infected P. kudriavzevii fungus in lemon, malta and  apple fruits showed a typical brown rot disease symptoms and the isolates showed similar size of  650bp clear band for PCR products. In antifungal activity assay, the methanol extracts of Allium  sativum displayed highest 17.18±0.66 mm diameter zone of inhibition against the isolated fungus.  Rhizobium leguminosarum and Alternaria spp. showed significant antagonistic effect in coculture  with the isolated fungus. The present research work would be helpful to control the  devastating brown rot disease of Citrus sinensis.


Keywords: Citrus sinensis, BRD, ITS, Pichia kudriavzevii, Antagonism

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