Dataset Information

Correlation of isolate-specific oxidative stress responses at the protein level in Aspergillus flavus with aflatoxin production capability

ABSTRACT: Aspergillus flavus is a pathogen of corn, peanut, and other crops which produces carcinogenic mycotoxins known as aflatoxins. Previous studies have shown that drought stress results in exacerbated aflatoxin production. Drought-associated oxidative stress caused by reactive oxygen species (ROS) is suspected to contribute to increased aflatoxin production during infection. Here, the responses of field isolates of A. flavus with varying degrees of aflatoxin production capability to H2O2-derived oxidative stress were examined using iTRAQ proteomics. Three isolates: AF13 (highly toxigenic), NRRL3357 (moderately toxigenic), and K54A (atoxigenic), were cultures in aflatoxin conducive yeast extract-sucrose (YES) medium amended with 0, 10, or 20/25mM of H2O2. Identified differentially expressed proteins were used for functional prediction, cellular localization, and pathway analyses to identify molecular mechanisms involved in A. flavus oxidative stress responses relative to aflatoxin production capability. Correlative analyses with previously obtained transcriptome data for the same isolates under the same experimental conditions was also performed with a low degree of correlation (r = 0.1114) observed between the protein and transcript data suggesting possible post-transcriptional regulation of oxidative stress responses. The identified stress responsive mechanisms provide a basis of investigating novel approaches of enhancing host resistance against aflatoxin contamination.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Aspergillus Flavus

TISSUE(S): Sclerotium, Fungal Cell, Conidium, Mycelium

SUBMITTER: Jake Fountain

LAB HEAD: Baozhu Guo

PROVIDER: PXD007164 | Pride | 2018-02-27

REPOSITORIES: Pride

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Proteome analysis of Aspergillus flavus isolate-specific responses to oxidative stress in relationship to aflatoxin production capability.

Fountain Jake C JC Koh Jin J Yang Liming L Pandey Manish K MK Nayak Spurthi N SN Bajaj Prasad P Zhuang Wei-Jian WJ Chen Zhi-Yuan ZY Kemerait Robert C RC Lee R Dewey RD Chen Sixue S Varshney Rajeev K RK Guo Baozhu B

Scientific reports 20180221 1

Aspergillus flavus is an opportunistic pathogen of plants such as maize and peanut under conducive conditions such as drought stress resulting in significant aflatoxin production. Drought-associated oxidative stress also exacerbates aflatoxin production by A. flavus. The objectives of this study were to use proteomics to provide insights into the pathogen responses to H<sub>2</sub>O<sub>2</sub>-derived oxidative stress, and to identify potential biomarkers and targets for host resistance breedin ...[more]

PMID: 29467403

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Project description:Aspergillus flavus is the major producer of carcinogenic aflatoxins in crops worldwide. Natural populations of A. flavus show tremendous variation in aflatoxin production some of which can be attributed to extreme environmental conditions (e.g., drought), differential regulation of the aflatoxin biosynthetic pathway, missing cluster genes or loss-of-function mutations. Understanding the evolutionary processes that generate genetic diversity in A. flavus may also explain quantitative and qualitative differences in aflatoxigenicity. Several population studies provide indirect evidence of recombination in the aflatoxin gene cluster and genome-wide, using multilocus genealogical approaches. More recently A. flavus has been shown to be functionally heterothallic and capable of sexual reproduction in laboratory crosses. In the present study, we characterize the progeny from nine A. flavus crosses and show that crossovers in the aflatoxin cluster coincide with inferred recombination blocks and hotspots in natural populations, which suggests that recombination in the cluster is primarily driven by sex. Moreover, we show that a single crossover event in the cluster can restore aflatoxigenicity, which is significant as mycotoxin production in A. flavus is highly heritable. aCGH was used to corroborate inferences from cluster-based MLSTs and to possibly identify additional crosovers within the cluster. aCGH comparison between 3 strains of A. flavus: 2 parental (P) and 1 progeny (F1) analyzed at the probe level. A total of 9 trio comparisons were made from a total of 18 isolates analyzed by aCGH. Trio comparisons are as follows: IC278 (P), IC1179 (P) and IC1650 (F1); IC201 (P), IC310 (P) and IC1719 (F1); IC307 (P), IC308 (P) and IC1751 (F1); IC277 (P), IC311 (P) and IC1766 (F1); IC277 (P), IC311 (P) and IC1775 (F1); IC244 (P), IC277 (P) and IC2205 (F1); IC244 (P), IC277 (P) and IC2207 (F1); IC301 (P), IC1179 (P) and IC2171 (F1); and finally IC244 (P), IC277 (P) and IC2209 (F1).

Project description:Aflatoxins are carcinogenic fungal secondary metabolites. Levels of aflatoxins in agricultural commodities are stringently regulated by many countries. A cluster of genes is responsible for aflatoxin biosynthesis by Aspergillus flavus and other closely related species. Expression of the clustered aflatoxin genes is governed by a complex network of regulatory mechanisms. To better understand the molecular events that are associated with aflatoxin production, transcription profiling by microarray analyses which compared three independent aflatoxigenic A. flavus strains to individual isogenic progenies that no longer produced aflatoxins after serial transfers was carried out. Twenty-two significantly differentially expressed features were identified. After physical mapping using the A. oryzae genome sequence as the reference, the number of unique genes was reduced to 16. Compared to the parental strains, changes in the aflatoxin gene expression levels in the progenies were not significant, which suggests that the inability to produce aflatoxins is not caused by decreased expression. The only gene showing higher expression levels in the progenies is homologous to glutathione S-transferease genes. Overexpression of this gene, named hcc, at six- to nine-fold in an aflatoxigenic A. flavus did not cause discernible changes in colony morphology or aflatoxin production. Loss of aflatoxin production after serial transfers may not result from a single event but caused by multiple factors. Keywords: Compartiave hybridization toxigenic and atoxigenic lines of Aspergillus Aspergillus flavus NRRL 29459, NRRL 29474, and NRRL 29490 are aflatoxigenic strains originated from soil collection in a peanut field (Terrell Co., Georgia, USA). Strains 459B-20-2, 474A-20, and 499A-20 were nonaflatoxigenic isolates obtained after 20 serial transfers of the parental strains on potato dextrose agar slants (Horn and Dorner 2002). Comparsions in each experiment consisted of one aflatoxigenic parental strain and one nonaflatoxigenic progeny, compared after 48- or 72-hr growth. Each comparison was repeated with duplicate dye-flip.

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