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Purification of the Antifungal Protein from Antagonistic Bacterium BDT-25 Against Verticillium Dahliae

Author: SunYao
Tutor: ZhuBaoCheng;MaPing
School: Hebei University
Course: Botany
Keywords: Verticillium dahilae Kleb antagonistic bacterium antifungal protein purification
CLC: S435.621
Type: Master's thesis
Year: 2007
Downloads: 15
Quote: 0
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Abstract


Antagonistic bacterial strain BDT-25 was isolated from the rhizosphere of cotton. Antagonistic test showed that the strain showed significant inhibitory effect on mycelium growth and microsclerotia formation of Verticillium dahilae Kleb. Further inhibition tests of strain BDT-25 were conducted on different plant fungal pathogens. Results showed that strain BDT-25 also showed significant antagonistic to other 10 pathogens including Fusarium oxysporium f.sp.vasinfectum, Rhizoctonia solani et al. Identification of strain BDT-25 was carried out through physiological and biochemical testing, the result confirmed that it is Bacillus subtilis.Antagonistic activities of strain BDT-25 were evaluated under different culture conditions. The result showed that the strain could produce more antifungal protein under the optimum condition (cultured in NB broth at 170rpm for 48 hours under 28℃and, pH7.0).Precipitates of the fermentation liquid from strain BDT-25 were collected by 80% saturation ammonium sulfate and showed significant inhibitory effect on mycelium growth of V. dahilae, while the extraction of the fermentation liquid by organic solvents did not show any antagonistic activity. The antifungal substance from strain BDT-25 was sensitive to heat, Proteinases K, Trypsin and Pepsin, but was not sensitive to RNAase. According to the result, the antifungal substance could be approved antifungal protein.When the antifungal protein was purified by chromatography, four peaks were obtained from DEAE-Sepharose Fast the Flow, the fourth peak showed antifungal activity. After purified through Sephacry S-100, one of the two peaks named P1 showed antagonistic effect against V. dahliae. One single band was collected through SDS-PAGE and the molecular weight was about 30 kDa.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Crop pests and diseases and their prevention > Economic crop pests and diseases > Fiber crop pests and diseases > Cotton pests and diseases > Disease
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