Reduced fertilization supplemented with Bacillus safensis RGM 2450 and Bacillus siamensis RGM 2529 promotes tomato production in a sustainable way.

The rising demand for vegetables has driven the adoption of greenhouse cultivation to guarantee high yields and quality of fresh produce year-round. Consequently, this elevates the demand for fertilizers, whose costs are progressively escalating. Bacillus safensis RGM 2450 and Bacillus siamensis RGM 2529 are plant growth-promoting rhizobacteria (PGPR). The combination of these strains exhibited synergistic activity in stimulating the growth and seedling hydration of tomatoes. In this study, the effects of inoculation with a RGM 2450 plus RGM 2529 formulation were evaluated under 66% and 100% fertilization programs in tomato crops under greenhouse conditions. Fertilization programs (66% and 100%) with or without commercial biostimulants were used as control treatments. In this assay, the NPK percentage in the plant tissue, tomato average weight, tomato average weight per harvest, tomato diameter, and changes in the colonization, structure, and diversity of the bacterial rhizosphere were measured. The 100% and 66% fertilization programs supplemented with the RGM 2529 plus RGM 2450 formulation increased the average weight of tomatoes per harvest without statistical difference between them, but with the other treatments. The 66% fertilization with RGM 2450 plus RGM 2529 increased between 1.5 and 2.0 times the average weight of tomatoes per harvest compared to the 66% and 100% fertilizations with and without commercial biostimulant treatments, respectively. This study represents the first report demonstrating that the application of a formulation based on a mixture of B. siamensis and B. safensis in a fertilization program reduced by 33% is equivalent in productivity to a conventional fertilization program for tomato cultivation, achieving an increase in potential plant growth-promoting rizobacteria of the genus Flavobacterium. Therefore, the adoption of a combination of these bacterial strains within the framework of a 66% inorganic fertilization program is a sustainable approach to achieving greater tomato production and reducing the environmental risks associated with the use of inorganic fertilization.

Identification of Exoenzymes Secreted by Entomopathogenic Fungus Beauveria pseudobassiana RGM 2184 and Their Effect on the Degradation of Cocoons and Pupae of Quarantine Pest Lobesia botrana.

Beauveria pseudobassiana RGM 2184 has shown 80% maximum efficacy against the pest Lobesia botrana in the autumn and winter seasons. This suggests that the strain possesses an interesting battery of enzymes that are cold-adapted to penetrate the thick and hydrophobic cocoon of L. botrana. In this study, screening of the proteolytic, lipolytic, and chitinolytic activity of enzyme extracts secreted by the RGM 2184 strain was carried out in various culture media. The enzyme extracts with the highest activity were subjected to zymography and mass spectrometry. These analyses allowed the identification of two proteases, two lipases, and three chitinases. Comparative analysis indicated that the degree of similarity between these enzymes was substantially reduced when the highest degree of taxonomic relatedness between RGM 2184 and the entomopathogenic fungus strain was at the family level. These results suggest that there is a wide variety of exoenzymes in entomopathogenic fungi species belonging to the order Hypocreales. On the other hand, exoenzyme extract exposure of cocoons and pupae of L. botrana provoked damage at 10 °C. Additionally, an analysis of the amino acid composition of the RGM 2184 exoenzyme grouped them close to the cold-adapted protein cluster. These results support the use of this strain to control pests in autumn and winter. Additionally, these antecedents can form a scaffold for the future characterization of these exoenzymes along with the optimization of the strain's biocontrol ability by overexpressing them.

Genomic and Experimental Analysis of the Biostimulant and Antagonistic Properties of Phytopathogens of Bacillus safensis and Bacillus siamensis.

The B. safensis RGM 2450 and B. siamensis RGM 2529 strains were isolated from the rhizosphere of plants presenting resilience to abiotic and biotic stress conditions. To understand the implications of bacteria in resilience, a genomic and experimental analysis was carried out on their biostimulant and phytopathogenic antagonist properties. Genome analyses of both strains indicated that they have the potential to synthesize bioactive compounds such as the battery of non-ribosomal peptides, polyketides, extracellular enzymes and phytohormones. These results were consistent with the antagonistic activities of both strains against the phytopathogens Botrytis cinerea, Colletotrichum acutatum, Fusarium oxysporum and Phytophtora cinnamomi. They also showed the capacity to solubilize phosphorus, fix nitrogen and produce indole acetic acid. This was observed in tomato seedlings grown from seeds inoculated with the mixture of strains which presented significantly greater length as well as wet and dry weight in comparison with the treatments individually inoculated with each strain and the control. Accordingly, the combination of B. safensis RGM 2450 and B. siamensis RGM 2529 showed synergistic biostimulant activity. These findings contribute new knowledge of the genomic and metabolomic properties taking part in the symbiotic interactions between these strains and the plants and uphold the combined use of both strains as a biostimulant.

Genomic and Experimental Analysis of the Insecticidal Factors Secreted by the Entomopathogenic Fungus Beauveria pseudobassiana RGM 2184.

The entomopathogenic fungus Beauveria pseudobassiana strain RGM 2184 can reach a maximum efficacy of 80% against the quarantine pest Lobesia botrana in field assays. In this study, the RGM 2184 genome was sequenced, and genome mining analyses were performed to predict the factors involved in its insecticidal activity. Additionally, the metabolic profiling of the RMG 2184 culture's supernatants was analyzed by mass spectrometry, and the insecticidal activity from one of these extracts was evaluated in Galleria mellonella larvae. The genome analysis resulted in 114 genes encoding for extracellular enzymes, four biosynthetic gene clusters reported as producers of insecticidal and bactericidal factors (oosporein, beauvericin, desmethylbassianin, and beauveriolide), 20 toxins, and at least 40 undescribed potential biocontrol factors (polyketides and nonribosomal peptides). Comparative genomic analysis revealed that 65-95% of these genes are Beauveria genus-specific. Metabolic profiling of supernatant extracts from RGM 2184 cultures exhibited secondary metabolites such as beauveriolide, oosporein, inflatin C, and bassiatin. However, a number of detected metabolites still remain undescribed. The metabolite extract caused 79% mortality of Galleria mellonella larvae at 28 days. The results of this research lay the groundwork for the study of new insecticidal molecules.