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Comparative proteomics provides insights on the basis of resistance to aflatoxin production in peanut (Arachis hypogea L.)

ABSTRACT: Aflatoxins are highly toxic secondary metabolites produced mainly by Aspergillus flavus and A. parasiticus, which colonize a wide variety of food commodities especially under dry and hot conditions. We developed transgenic peanut expression four RNAi genes NsdC, Vea, Ver1 and aflR, by Agrobacterium-mediated transformation. To understand the proteome changes in 4RNAi and WT control lines, a label-free quantitative proteomics analysis was performed at 0, 30, 48 and 72 h after A. flavus inoculation using UPLC-ESI-MS/MS. Several resistance proteins in the secondary metabolic pathways related to phenylpropanoids, flavonoids, and fatty acid biosynthesis were strongly induced in the resistant genotype.

ORGANISM(S): Arachis Hypogaea

SUBMITTER: Dr Pooja Bhatnagar-Mathur

PROVIDER: PXD028196 | JPOST Repository | Thu Sep 02 00:00:00 BST 2021

REPOSITORIES: jPOST

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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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Comparative proteomics provides insights on the basis of resistance to aflatoxin production in peanut (Arachis hypogea L.)

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