Aflatoxin B₁ is a potent carcinogen produced by Aspergillus flavus, mainly during grain storage. As pre-harvest methods are insufficient to avoid mycotoxin presence during storage, diverse curative techniques are being investigated for the inhibition of fungal growth and aflatoxin detoxification. Streptomyces spp. represent an alternative as they are a promising source of detoxifying enzymes. Fifty-nine Streptomyces isolates and a Streptomyces griseoviridis strain from the commercial product Mycostop^®, evaluated against Penicillium verrucosum and ochratoxin A during previous work, were screened for their ability to inhibit Aspergillus flavus growth and decrease the aflatoxin amount. The activities of bacterial cells and cell-free extracts (CFEs) from liquid cultures were also evaluated. Fifty-eight isolates were able to inhibit fungal growth during dual culture assays, with a maximal reduction going down to 13% of the control. Aflatoxin-specific production was decreased by all isolates to at least 54% of the control. CFEs were less effective in decreasing fungal growth (down to 40% and 55% for unheated and heated CFEs, respectively) and aflatoxin-specific production, with a few CFEs causing an overproduction of mycotoxins. Nearly all Streptomyces isolates were able to degrade AFB₁ when growing in solid and liquid media. A total degradation of AFB₁ was achieved by Mycostop^® on solid medium, as well as an almost complete degradation by IX20 in liquid medium (6% of the control). CFE maximal degradation went down to 37% of the control for isolate IX09. The search for degradation by-products indicated the presence of a few unknown molecules. The evaluation of residual toxicity of the tested isolates by the SOS chromotest indicated a detoxification of at least 68% of AFB₁’s genotoxicity.

Ochratoxin A (OTA) is a secondary metabolite produced by fungal pathogens such as Penicillium verrucosum, which develops in food commodities during storage such as cereals, grapes, and coffee. It represents public health concerns due to its genotoxicity, carcinogenicity, and teratogenicity. The objective of this study was to evaluate the ability of actinobacteria and their metabolites to degrade OTA and/or to decrease its production. Sixty strains of actinobacteria were tested for their ability to prevent OTA formation by in vitro dual culture assays or with cell free extracts (CFEs). In dual culture, 17 strains strongly inhibited fungal growth, although it was generally associated with an increase in OTA specific production. Seventeen strains inhibited OTA specific production up to 4% of the control. Eleven actinobacteria CFEs reduced OTA specific production up to 62% of the control, while no substantial growth inhibition was observed except for two strains up to 72% of the control. Thirty-three strains were able to degrade OTA almost completely in liquid medium whereas only five were able to decrease it on solid medium, and two of them reduced OTA to an undetectable amount. Our results suggest that OTA decrease could be related to different strategies of degradation/metabolization by actinobacteria, through enzyme activities and secretion of secondary metabolites interfering with the OTA biosynthetic pathway. CFEs appeared to be ineffective at degrading OTA, raising interesting questions about the detoxification mechanisms. Common degradation by-products (e.g., OTα or L-β-phenylalanine) were searched by HPLC-MS/MS, however, none of them were found, which implies a different mechanism of detoxification and/or a subsequent degradation into unknown products.

Wheat production worldwide faces numerous challenges linked to climate change, exponential population growth, nutrient depletion in agricultural soils, and the increasing threat of phytopathogen occurrence. The application of beneficial microorganisms is a promising strategy for crop management as it favorizes nutrient uptake, improves soil fertility, and increases plant resilience. Therefore, this approach facilitates the transition to more sustainable agricultural practices while reducing the dependence on agrochemicals. The valuable beneficial impacts of bioinoculant application include the enrichment of agricultural soils’ ecosystems by restoring microbial populations and interactions that have been lost through the years due to decades of intensive agricultural practices and the massive application of pesticides. Furthermore, beneficial microorganisms constitute a remarkable tool for combating biotic threats, specifically fungal pathogens, whose proliferation and emergence are predicted to increase due to global warming. To optimize their beneficial impact, bioinoculant development requires an extensive study of microbial interactions with plants and their surrounding ecosystem, to improve their composition, mode of action, and stability through application. The use of innovative tools, such as omic sciences, facilitates the elucidation of these mechanisms. Finally, bioprospection and bioformulation must be consciously executed to guarantee the application and persistence of adapted microorganisms and/or their bioactive molecules.

Strain TE5 was isolated from a wheat (Triticum turgidum L. subsp. durum) rhizosphere grown in a commercial field of wheat in the Yaqui Valley in Mexico. In this work, we present strain TE5 as a promising biological control agent against Bipolaris sorokiniana. First, after its genome sequencing through Illumina NovaSeq, this strain showed a genome size of 4,262,927 bp, with a 43.74% G + C content, an N50 value of 397,059 bp, an L50 value of 4 bp, and 41 contigs (>500 bp). Taxonomical affiliation was carried out by using overall genome relatedness indexes (OGRIs) and the construction of a phylogenomic tree based on the whole genome. The results indicated that strain TE5 identifies with Bacillus cabrialesii subsp. cabrialesii. Genomic annotation using Rapid Annotation Using Subsystems Technology (RAST) and Rapid Prokaryotic Genome Annotation (Prokka) indicated the presence of 4615 coding DNA sequences (CDSs) distributed across 330 subsystems, which included gene families associated with biocontrol, stress response, and iron competition. Furthermore, when the antiSMASH 7.1 platform was used for genome mining, the results indicated the presence of seven putative biosynthetic gene clusters related to the production of biocontrol metabolites, namely subtilosin A, bacillibactin, fengycin, bacillaene, bacilysin, surfactin, and rhizocticin A. Moreover, the antifungal activity of strain TE5 and its cell-free extract (CFE) was evaluated against Bipolaris sorokiniana, an emergent wheat pathogen. The results of in vitro dual confrontation showed fungal growth inhibition of 67% by strain TE5. Additionally, its CFE almost completely inhibited (93%) the growth of the studied phytopathogenic fungus on liquid media. Further observations of the impact of these bacterial metabolites on fungal spore germination exhibited inhibition of fungal spores through degrading the germinative hypha, avoiding mycelium development. Finally, the protective effect of strain TE5 against Bipolaris sorokiniana was evaluated for wheat seedlings. The results showed a significant decrease (83%) in disease severity in comparison with the plant infection without inoculation of the biological control agent. Thus, this work proposes Bacillus cabrialesii subsp. cabrialesii strain TE5 as a promising biological control agent against the wheat pathogen Bipolaris sorokiniana while suggesting lipopeptides as the potential mode of action, together with plant growth and defense stimulation.

Bacterial strain TSO55 was isolated from a commercial field of wheat (Triticum turgidum L. subsp. durum), under organic amendments, located in the Yaqui Valley, Mexico. Morphological and microscopical characterization showed off-white irregular colonies and Gram-positive bacillus, respectively. The draft genome sequence of this strain revealed a genomic size of 5,489,151 bp, with a G + C content of 35.21%, N50 value of 245,934 bp, L50 value of 8, and 85 contigs. Taxonomic affiliation showed that strain TSO55 belongs to Bacillus paranthracis, reported as an emergent human pathogen. Genome annotation identified 5743 and 5587 coding DNA sequences (CDSs), respectively, highlighting genes associated with indole production, phosphate and potassium solubilization, and iron acquisition. Further in silico analysis indicated the presence of three CDSs related to pathogenicity islands and a high pathogenic potential (77%), as well as the presence of multiple gene clusters related to antibiotic resistance. The in vitro evaluation of plant growth promotion traits was negative for indole production and phosphate and potassium solubilization, and it was positive but low (18%) for siderophore production. The biosynthetic gene cluster for bacillibactin (siderophore) biosynthesis was confirmed. Antifungal bioactivity of strain TSO55 evaluated against wheat pathogenic fungi (Alternaria alternata TF17, Bipolaris sorokiniana TPQ3, and Fusarium incarnatum TF14) showed minimal fungal inhibition. An antibiotic susceptibility assay indicated resistance to three of the six antibiotics evaluated, up to a concentration of 20 µg/mL. The beta hemolysis result on blood agar reinforced TSO55’s pathogenic potential. Inoculation of B. paranthracis TSO55 on wheat seedlings resulted in a significant decrease in root length (−8.4%), total plant height (−4.2%), root dry weight (−18.6%), stem dry weight (−11.1%), and total plant dry weight (−15.2%) compared to the control (uninoculated) treatment. This work highlights the importance of analyzing the microbiological safety of organic amendments before application. Comprehensive genome-based taxonomic affiliation and bioprospecting of microbial species introduced to the soil by organic agricultural practices and any microbial inoculant will prevent the introduction of dangerous species with non-beneficial traits for crops, which affect sustainability and generate potential health risks for plants and humans.