Project description:Lecanicillium fungicola, the causative agent of dry bubble disease on Agaricus bisporus results in significant crop losses for mushroom growers worldwide. Dry bubble disease is treated through strict hygiene control methods and the application of chemical fungicides but an increase in fungicide resistant L. fungicola strains has resulted in a need to develop alternative biocontrol treatment methods. The aim of the work presented here was to evaluate the response of L. fungicola to a Bacillus velezensis isolate to assess its potential as a novel biocontrol agent. The bacterial species in Serenade, a commercially available biocontrol treatment was also included in this analysis. Exposure of 48 hr L. fungicola cultures to 25% v/v 96h B. velezensis culture filtrate resulted in a 45% reduction in biomass (P < 0.0002) and a disruption in hyphal structure and morphology. Characterisation of the proteomic response of L. fungicola following exposure to B. velezensis culture filtrate revealed an increase in the abundance of a variety of proteins associated with stress response (Norsolorinic acid reductase (+8 fold), isocitrate lyase (+7 fold) and MMS19 nucleotide excision repair protein (+4 fold). There was also a decrease in the abundance of proteins associated with transcription (40S ribosomal protein S30 (-33 fold), 60S ribosomal protein L5 (-45 foldThe results presented here indicate that B. velezensis culture filtrate is capable of inhibiting the growth of L. fungicola and inducing a stress response, thus indicating its potential to control this important pathogen of mushrooms.
Project description:Two potato cultivars, Russet Burbank and Bionta, were inoculated with three different endophytes containing different AHL types. The impact of the endophytes to the different cultivars was measured by gene expression analysis with a customized microarray B. phytofirmans type strain PsJN was originally isolated as a contaminant from surface-sterilized, Glomus vesculiferum-infected onion roots (Nowak et al., 1998), whereas strain P6 RG6-12 was isolated from the rhizosphere of a grassland in the Netherlands (Salles et al., 2006). This strain was selected based on its similarity to strain PsJN based on 16S rRNA gene homology, and similar phenotypic features. Both strains were generally cultivated on King's medium (King et al., 1954). For the mutant AHL to the strain B. phytofirmans PsJN a quorum quenching approach as described by Wopperer et al., 2006 was employed. Plasmid pMLBAD-aiiA, which contains aiiA, the Bacillus sp. 240B1 lactonase gene, was transferred to B. phytofirmans PsJN by triparental mating as described by de Lorenzo and Timmis (1994). 2 cultivars, 3 endophytes
Project description:Cladobotryum mycophilum, the causative agent of cobweb disease on Agaricus bisporus results in significant crop losses for mushroom growers worldwide. Cobweb disease is treated through strict hygiene control methods and the application of chemical fungicides but an increase in fungicide resistant Cladobotryum strains has resulted in a need to develop alternative biocontrol treatment methods. The aim of the work presented here was to evaluate the response of C. mycophilum to a Bacillus velezensis isolate to assess its potential as a novel biocontrol agent. Exposure of 48 hr C. mycophilum cultures to 25% v/v 96h B. velezensis culture filtrate resulted in a 57% reduction in biomass (P < 0.0002), a disruption in hyphal structure and morphology, and the appearance of aurofusarin in culture medium. Proteomic analysis of B. velezensis culture filtrate revealed the presence of peptidase 8 (subtilisin), peptide deformylase and probable cytosol aminopeptidase which are known to induce cell disruption. Characterisation of the proteomic response of C. mycophilum following exposure to B. velezensis culture filtrate revealed an increase in the abundance of a variety of proteins associated with stress response (ISWI chromatin-remodelling complex ATPase ISW2 (+24 fold), carboxypeptidase Y precursor (+3 fold) and calmodulin (+2 fold). There was also a decrease in the abundance of proteins associated with transcription (40S ribosomal protein S30 (-26 fold), 40S ribosomal protein S21 (-3 fold) and carbohydrate metabolism, (L-xylulose reductase (-10 fold). The results presented here indicate that B. velezensis culture filtrate is capable of inhibiting the growth of C. mycophilum and inducing a stress response, thus indicating its potential to control this important pathogen of mushrooms.
Project description:The present study aims to evaluate the response of the three Mediterranean local grapevines ‘Garnacha Blanca’, ‘Garnacha Tinta’, and ‘Macabeo’ to treatments with biocontrol products (BPs), a botanical extract (Akivi, Dittrichia viscosa extract) and a beneficial microorganism (Bacillus UdG, Bacillus velezensis). A combination of transcriptomics and metabolomics approaches were chosen in order to study grapevine gene expression and to identify gene marker candidates, as well as, to determine grapevine metabolites differentially concentrated in response to BPs treatments. Grapevine plants were cultivated in greenhouse controlled conditions and submitted to the treatments, and thereafter, leaves were sampled 24h after treatment to conduct gene expression study by RNA-sequencing for ‘Garnacha Blanca’ leaves extract and by RT-qPCR for the three cultivars. Differentially expressed genes (DEGs) were investigated for both treatments and highly influenced DEGs were selected to be tested in the three cultivars as treatment gene markers. In addition, extraction of leaf components was performed to quantify metabolites such as phytohormones, organic acids, and phenols. Considering all the upregulated and downregulated genes and enhanced metabolites concentrations, the treatments had an effect on jasmonic acid, ethylene, and phenylpropanoids defense pathways. In addition, several DEG markers were identified presenting a stable overexpression after the treatments in the three grapevine cultivars. These gene markers could be used to monitor the activity of the products in field treatments in future research. Further research will be necessary to confirm these first results under field conditions.