Botrytis cinerea is the causal agent of gray mold, a disease that attacks many plants worldwide. This phytosanitary problem was reported in orchards in the State of Mexico, Mexico, specifically in pomegranate crops. This study isolated this phytopathogen from fruits collected in the municipality of Chilcuautla, state of Hidalgo, Mexico, indicating that the disease is spreading across the country. Two strains of phytopathogens, MIC and MID, were obtained after conducting Koch’s postulates. These strains were identified molecularly by amplifying and analyzing the concatenated sequence of the regions that correspond to the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (G3PDH), and DNA-dependent RNA polymerase subunit II (RPB2). In vitro confrontation tests demonstrated that Bacillus velezensis 160 inhibited the development of both strains of B. cinerea under study through the action of nonvolatile, thermostable, and volatile metabolites. Applying this bacterium to pomegranate orchards for two consecutive years (2022 and 2023) decreased the incidence, severity, and progression of the disease. We conclude that this bacterium can be used as a fungicide to control gray mold in pomegranate crops. [ ] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

The aim of this study was to determine biofilm production and motility in Bacillus pacificus B630 and Bacillus cereus ATCC 14579, and to perform a comparative genome analysis using bioinformatic tools to understand the differences between the two strains. Biofilm production was performed in glass tubes stained with safranin; motility was determined on soft agar. Bioinformatic analysis was performed using genomic information from both strains, including the identification of orthologous genes, the similarity between genes of the eps1 and sipW-tasA-calY operons, and the SipW and TasA model prediction. B. pacificus B630 produces a greater amount of biofilm on glass than B. cereus ATCC 14579 (p < 0.01). Furthermore, B. pacificus B630 shows lower motility than B. cereus ATCC 14579 (p < 0.001). B. pacificus B630 contains 45 unshared genes, whereas B. cereus ATCC 14579 has 27 unshared genes. Differences in similarity were observed between the genes of the eps1 and sipW-tasA-calY operons. These differences between SipW and TasA may affect protein structural predictions. In SipW, the differences may affect the C-terminal region. In TasA, the number of B-sheets differed between the two proteins, and amino acid substitutions were found in regions of high protein aggregation. Genomic differences in genes associated with biofilm production may explain differences in biofilm production between the strains studied.

Gray mold disease, caused by the fungus Botrytis cinerea, affects a wide variety of plants. In this study, we conducted several in vitro tests and genomic analyses on three strains of this fungus (BcPgIs-1, BcPgIs-3, MIC) previously isolated from diseased pomegranate fruits, collected at two geographic locations in Mexico. Our goal was to identify possible differences among these strains. The development of the three strains in distinct culture media, the production of extracellular enzymes, and their effect on the progression of infection in pomegranate fruits were evaluated. The genomes were sequenced using the Illumina platform and analyzed with various bioinformatics tools. All strains possess genetic determinants for virulence and cell wall polymer degradation, but MIC exhibited the highest pectinolytic activity in vitro. This strain also produced sclerotia in a shorter time (7 days) in PDA medium. BcPgls-3 demonstrated the highest conidia production across all the culture media used. Both BcPgls-3 and MIC damaged all the pomegranate fruits 8 days after inoculation, while the BcPgls-1 required up to 9 days. Sequencing of the three strains yielded high-quality sequences, resulting in a total of 17 scaffolds and genomes that exceed 41 million bp, with a GC content of approximately 42%. Phylogenomic analysis indicated that the MIC strain is situated in a group separate from BcPgIs-1 and BcPgIs-3. BcPgIs-3 possesses more coding sequences, but MIC has more genes for CAZymes and peptidases. The three strains share 10,174 genes, while BcPgIs-3 and MIC share 851. These findings highlight the differences among the strains studied, which may reflect their adaptive capacities to their environment. Results contribute to our understanding of the biology of gray mold in pomegranates and could assist in developing more effective control strategies.

Spontaneous fermentations contribute to the identity of ancestral mezcal, although duration and yield may vary. This study selected an indigenous yeast and evaluated it as a starter candidate. Four native isolates (Torulaspora delbrueckii A1GM and A6GM, Kluyveromyces marxianus A2GM, and Saccharomyces cerevisiae A5GM) and a spontaneous control were compared in Agave cupreata microfermentations. Selection considered distillate alcohol content and preliminary ethanol-tolerance assays. A5GM was subsequently evaluated in three production batches containing A. angustifolia + A. karwinskii, A. angustifolia, or A. cupreata + A. angustifolia, with three independent vats per treatment. Fermentation profiles, distillate composition, yield, and sensory acceptance were assessed. A5GM produced the highest microfermentation distillate alcohol content (43.91 ± 0.89% Alc. Vol.) and grew at 16% ethanol (v/v). In the first two production batches, inoculated vats showed higher volumes standardized to 45% Alc. Vol. and lower calculated agave requirements than the controls; these were batch-specific observations under the operating conditions studied. Sensory evaluation used pooled vat-derived distillate composites, and the A5GM composites received the highest mean hedonic scores. Because strain-level establishment and persistence of A5GM were not confirmed, the observed outcomes cannot be attributed exclusively to this strain. Sensory interpretation is limited by fixed presentation order. Further independent batches, standardized management, and strain-level monitoring are needed to assess reproducibility.

Spontaneous fermentation is central to ancestral mezcal production, yet the bacterial and fungal communities associated with Agave cupreata must remain insufficiently characterized. This exploratory study described community composition and predicted functional profiles at three stages of a traditional fermentation defined by sugar depletion: 10 °Brix (0 h), 5 °Brix (72 h), and 2 °Brix (144 h). Ambient temperature ranged from 21.7 to 23.7 °C, must temperature from 28.5 to 29.9 °C, pH from 4.6 to 5.2, and relative humidity from 51.3% to 58.7%. At each stage, material collected from two wooden vats and three vertical positions was pooled into one composite sample. Bacterial 16S rRNA gene and fungal ITS1 amplicons were sequenced and analyzed using QIIME 2. At 10 °Brix, five dominant bacterial genera accounted for more than 93% of relative abundance. Lactic and acetic acid bacteria represented more than 91% at 5 °Brix, while Pediococcus, Lactiplantibacillus, and Loigolactobacillus were prominent at 2 °Brix. Seven fungal genera represented more than 98% of the initial profile, whereas Saccharomyces, Debaryomyces, and Maudiozyma accounted for more than 98% at 2 °Brix. PICRUSt2 predicted pathways related to carbohydrate degradation and fermentation, whereas FUNGuild assignments were predominantly saprotrophic at the later stages. Because one pooled sample was analyzed per stage, these patterns are descriptive and require validation in replicated fermentations. Despite these limitations, this study provides baseline information on the microbial communities associated with ancestral A. cupreata fermentation and identifies taxa and predicted functions that merit future isolation and experimental validation.