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Biocontrol capability of local Metschnikowia sp. isolates.

ABSTRACT: This study set out to isolate and identify epiphytic yeasts producing pulcherrimin, and to evaluate their potential as biological control agents (BCAs). We isolated Metschnikowia sp. strains from flowers and fruits collected in Poland. The plant material had been collected between April to September 2017 from two small orchards where traditional organic management is employed. We identified the essential phenotypic features of the yeast, including assimilation and enzymatic profiles, stress resistance, adhesion properties, and antimicrobial activity against various fungi involved in crop and/or food spoilage. Yeast screening was performed using YPD agar supplemented with chloramphenicol and Fe(III) ions. Taxonomic classification was determined by sequence analysis of the D1/D2 domains of the large subunit rRNA gene. The isolates were identified as Metschnikowia andauensis and Metschnikowia sinensis. The yeast isolates were further characterized based on their enzymatic and assimilation profiles, as well as their growth under various stress conditions. In addition, the hydrophobicity and adhesive abilities of the Metschnikowia isolates were determined using a MATH test and luminometry. Their antagonistic action against molds representing typical crop spoiling microflora was also evaluated. The assimilation profiles of the wild isolates were similar to those displayed by collection strains of M. pulcherrima. However, some of the isolates displayed more beneficial phenotypic properties, especially good growth under stress conditions. Several of the epiphytes grew well over a wider range of temperatures (8-30 °C) and pH levels (3-9), and additionally showed elevated tolerance to ethanol (8%), glucose (30%), and peroxides (50 mM). The hydrophobicity and adhesion of the yeast cells were strain- and surface-dependent. The tested yeasts showed potential for use as BCAs, with some exhibiting strong antagonism against molds belonging to the genera Alternaria, Botrytis, Fusarium, Rhizopus, and Verticillium, as well as against yeasts isolated as food spoilage microbiota.

SUBMITTER: Pawlikowska E

PROVIDER: S-EPMC6748895 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Biocontrol capability of local Metschnikowia sp. isolates.

Pawlikowska Ewelina E James Steve A SA Breierova Emilia E Antolak Hubert H Kregiel Dorota D

Antonie van Leeuwenhoek 20190520 10

This study set out to isolate and identify epiphytic yeasts producing pulcherrimin, and to evaluate their potential as biological control agents (BCAs). We isolated Metschnikowia sp. strains from flowers and fruits collected in Poland. The plant material had been collected between April to September 2017 from two small orchards where traditional organic management is employed. We identified the essential phenotypic features of the yeast, including assimilation and enzymatic profiles, stress resi ...[more]

PMID: 31111331

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Project description:Metschnikowia persimmonesis, a novel endophytic yeast strain isolated from Diospyros kaki calyx, possesses strong antimicrobial activity. We investigated its potential use as an environmentally safe food biocontrol agent through genomics, transcriptomics, and metabolomics. Secondary metabolites were isolated from M. persimmonesis, followed by chemical structure elucidation, PUL gene cluster identification, and RNA sequencing. Pulcherrimin was isolated using 2 M NaOH, its structure was confirmed, and the yield was quantified. Biocontrol efficacy of M. persimmonesis on persimmon fruits and calyx was evaluated by assessing lesion diameter and disease incidence. Following compounds were isolated from M. persimmonesis co-culture with Botrytis cinerea and Fusarium oxysporum: fusaric acid, benzoic acid, benzeneacetic acid, 4-hydroxybenzeneacetic acid, 4-(-2-hydoxyethyl)-benzoic acid, cyclo (Leu-Leu), benzenemethanol, 4-hydroxy-benzaldehide, 2-hydroxy-4-methoxy-benzoic acid, 4-hydroxy-benzoic acid, lumichrome, heptadecanoic acid, and nonadecanoic acid. Exposing M. persimmonesis to different growth media conditions (with or without sugar) resulted in the isolation of five compounds: Tyrosol, Cyclo (Pro-Val), cyclo(L-Pro-L-Tyr), cyclo(Leu-Leu), and cyclo(l-tyrosilylicine). Differentially expressed gene analysis revealed 3264 genes that were significantly expressed (fold change ≥2 and p-value ≤0.05) during M. persimmonesis growth in different media, of which only 270 (8.27%) showed altered expression in all sample combinations with Luria-Bertani Agar as control. Minimal media with ferric ions and tween-80 triggered the most gene expression changes, with the highest levels of PUL gene expression and pulcherrimin yield (262.166 mg/L) among all media treatments. M. persimmonesis also produced a higher amount of pulcherrimin (209.733 mg/L) than Metschnikowia pulcherrima (152.8 mg/L). M. persimmonesis inhibited the growth of Fusarium oxysporum in persimmon fruit and calyx. Toxicity evaluation of M. persimmonesis extracts showed no harmful effects on the liver and mitochondria of zebrafish, and no potential risk of cardiotoxicity in hERG-HEK293 cell lines. Thus, M. persimmonesis can be commercialized as a potent and safe biocontrol agent for preserving food products.

Dataset Information

Biocontrol capability of local Metschnikowia sp. isolates.

Publications

Biocontrol capability of local Metschnikowia sp. isolates.

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