Control of Aspergillus carbonarius in grape berries by Lactobacillus plantarum: A phenotypic and gene transcription study

Abstract

The in vitro and in situ antifungal activity of Lactobacillus plantarum against the ochratoxigenic fungus Aspergillus carbonarius was investigated in this study. Four different fungal isolates from grape berries were co-cultured with four different strains of L. plantarum on Malt Extract Agar (MEA) plates at 30 °C. Bacterial strains inhibited fungal growth up to 88% and significantly reduced toxin production up to 100%. In addition, L. plantarum was evaluated as biocontrol agent against A. carbonarius growth and OTA production on table grapes. Temporal studies of bacterial antagonism were performed with two different grape cultivars. Artificially wounded and unwounded berries were pre-treated with 10⁸ CFU/mL bacteria and inoculated with 10⁶ spores/mL of A. carbonarius ochratoxigenic isolates. Biocontrol agents displayed high rate of colonization on grapes during 5 days of incubation at 30 °C. Scanning electron microscopy (SEM) also determined the presence of microorganisms on grape surface. Bacterial strains were effective in controlling fungal infection reaching up to 71% inhibition rates. However the presence of wounds on grape skin facilitated infection of berries by A. carbonarius, since unwounded berries showed lower levels of infection. Results also revealed significant reduction in mycotoxin production ranging between 32% and 92%. Transcriptome analysis following exposure to co-cultivation, exhibited differential expression for each gene studied of AcOTAnrps (Aspergillus carbonarius OTA nonribosomal), AcOTApks (Aspergillus carbonarius OTA polyketide synthase) and laeA, emphasizing the significance of strain variability. The genes AcOTAnrps and laeA were most influenced by the presence of L. plantarum. This work is a contribution for the potential biocontrol of toxigenic fungi in table grapes by lactic acid bacteria (LAB). The above findings underline the significance of bacterial strain variability on the effectiveness of biopreservative features of L. plantarum strains.

Keywords: Aspergillus carbonarius; Biocontrol; Grapes; Lactobacillus plantarum; Ochratoxin A; Transcriptomics.