Dataset Information

Antifungal Activity of an Abundant Thaumatin-Like Protein from Banana against Penicillium expansum, and Its Possible Mechanisms of Action.

ABSTRACT: Thaumatin-like protein from banana (designated BanTLP) has been purified by employing a simple protocol consisting of diethylaminoethyl Sephadex (DEAE?Sephadex) chromatography, gel filtration on Sephadex G50, and reversed-phase chromatography. The purified protein was identified by MALDI-TOF mass spectrometry, with an estimated molecular weight of 22.1 kDa. BanTLP effectively inhibited in vitro spore germination of Penicillium expansum, one of the main postharvest pathogens in fruits. This study further investigated the antifungal properties and underlying mechanisms of BanTLP against P. expansum. Results demonstrated that BanTLP exhibited antifungal activity in a wide pH range (4.0?10.0) at 20?50 °C. Propidium iodide (PI) influx and potassium release confirmed that BanTLP induced membrane disruption of the test pathogen, increasing the membrane permeability and disintegration of the cell. This led to cell death, as evidenced by the assays of thiobarbituric acid-reactive species (TBARS) content, the production of reactive oxygen species (ROS), and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence integrity. Ultrastructural alterations in P. expansum conidia after BanTLP treatment revealed severe damage to the cell wall. These results suggest that BanTLP purified from banana exerts antifungal activity against P. expansum by inducing plasma membrane disturbance and cell wall disorganization.

SUBMITTER: Jiao W

PROVIDER: S-EPMC6099679 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Antifungal Activity of an Abundant Thaumatin-Like Protein from Banana against <i>Penicillium expansum</i>, and Its Possible Mechanisms of Action.

Jiao Wenxiao W Li Xiangxin X Zhao Handong H Cao Jiankang J Jiang Weibo W

Molecules (Basel, Switzerland) 20180614 6

Thaumatin-like protein from banana (designated BanTLP) has been purified by employing a simple protocol consisting of diethylaminoethyl Sephadex (DEAE⁻Sephadex) chromatography, gel filtration on Sephadex G50, and reversed-phase chromatography. The purified protein was identified by MALDI-TOF mass spectrometry, with an estimated molecular weight of 22.1 kDa. BanTLP effectively inhibited in vitro spore germination of <i>Penicillium expansum</i>, one of the main postharvest pathogens in fruits. Thi ...[more]

PMID: 29899211

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Project description:Penicillium expansum causes blue mold, the most economically important postharvest disease of pome fruit worldwide. Beside sanitation practices, the disease is managed through fungicide applications at harvest. Difenoconazole (DIF) is a new demethylation inhibitor (DMI) fungicide registered recently to manage postharvest diseases of pome fruit. Herein, we evaluated the sensitivity of 130 P. expansum baseline isolates never exposed to DIF and determined the effective concentration (EC50) necessary to inhibit 50% germination, germ tube length, and mycelial growth. The respective mean EC50 values of 0.32, 0.26, and 0.18 μg/ml indicate a high sensitivity of P. expansum baseline isolates to DIF. We also found full and extended control efficacy in vivo after 6 months of storage at 1°C. We conducted a risk assessment for DIF-resistance development using ultraviolet excitation combined with or without DIF-selection pressure to generate and characterize lab mutants. Fifteen DIF-resistant mutants were selected and showed EC50 values of 0.92 to 1.4 μg/ml and 1.7 to 3.8 μg/ml without and with a DIF selection pressure, respectively. Resistance to DIF was stable in vitro over a 10-week period without selection pressure. Alignment of the full CYP51 gene sequences from the three wild-type and 15 mutant isolates revealed a tyrosine to phenylalanine mutation at codon 126 (Y126F) in all of the 15 mutants but not in the wild-type parental isolates. Resistance factors increased 5 to 15-fold in the mutants compared to the wild-type-isolates. DIF-resistant mutants also displayed enhanced CYP51 expression by 2 to 14-fold and was positively correlated with the EC50 values (R 2 = 0.8264). Cross resistance between DIF and fludioxonil, the mixing-partner in the commercial product, was not observed. Our findings suggest P. expansum resistance to DIF is likely to emerge in commercial packinghouse when used frequently. Future studies will determine whether resistance to DIF is qualitative or quantitative which will be determinant in the speed at which resistance will develop and spread in commercial packinghouses and to develop appropriate strategies to extend the lifespan of this new fungicide.

Dataset Information

Antifungal Activity of an Abundant Thaumatin-Like Protein from Banana against Penicillium expansum, and Its Possible Mechanisms of Action.

Publications

Antifungal Activity of an Abundant Thaumatin-Like Protein from Banana against <i>Penicillium expansum</i>, and Its Possible Mechanisms of Action.

Similar Datasets

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