Co-inoculation of the endophytes Bacillus thuringiensis CAPE95 and Paenibacillus polymyxa CAPE238 promotes Tropaeolum majus L. growth and enhances its root bacterial diversity.

Tropaeolum majus L. is a versatile edible plant that is widely explored due to its medicinal properties and as a key element in intercropping systems. Its growth could be improved by the use of biofertilizers that can enhance nutrient uptake by the plant or provide tolerance to different abiotic and biotic stresses. In a previous study, 101 endophytes isolated from T. majus roots showed more than three plant growth-promoting (PGP) features in vitro, such as phosphate mineralization/solubilization, production of siderophores, antimicrobial substances and indole-related compounds, and presence of the nifH gene. To provide sustainable alternatives for biofertilization, the genomes of two promising endophytes-CAPE95 and CAPE238-were sequenced to uncover metabolic pathways related to biofertilization. Greenhouse experiments were conducted with 216 seeds and 60 seedlings, half co-inoculated with the endophytes (treatment) and half inoculated with 1X PBS (control), and the impact of the co-inoculation on the plant's bacteriome was accessed through 16S rRNA gene metabarcoding. The strains CAPE95 and CAPE238 were taxonomically assigned as Bacillus thuringiensis and Paenibacillus polymyxa, respectively. Metabolic pathways related to the enhancement of nutrient availability (nitrogen fixation, sulfate-sulfur assimilation), biosynthesis of phytohormones (indole-3-acetic acid precursors) and antimicrobial substances (bacilysin, paenibacillin) were found in their genomes. The in vivo experiments showed that treated seeds exhibited faster germination, with a 20.3% higher germination index than the control on the eleventh day of the experiment. Additionally, treated seedlings showed significantly higher plant height and leaf diameters (p < 0.05). The bacterial community of the treated plants was significantly different from that of the control plants (p < 0.001) and showed a higher richness and diversity of species (Chao and Shannon indexes, p < 0.001). A higher relative abundance of potential synergistic PGP bacteria was also shown in the bacteriome of the treated plants, such as Lysinibacillus and Geobacter. For the first time, co-inoculation of B. thuringiensis and P. polymyxa was shown to have great potential for application as a biofertilizer to T. majus plants. The bacterial consortium used here could also be explored in other plant species in the future.

Fighting Obesity-Related Micronutrient Deficiencies through Biofortification of Agri-Food Crops with Sustainable Fertilization Practices.

Obesity is a critical medical condition worldwide that is increasingly involved with nutritional derangements associated with micronutrient deficiencies, including iron, zinc, calcium, magnesium, selenium, and vitamins A, C, D, and E. Nutritional deficiencies in obesity are mainly caused by poor-quality diets, higher nutrient requirements, alterations in micronutrient metabolism, and invasive obesity treatments. The current conventional agricultural system is designed for intensive food production, focusing on food quantity rather than food quality, consuming excessive agricultural inputs, and producing nutrient-deficient foods, thus generating severe health and environmental problems; agricultural food products may worsen obesity-related malnutrition. Therefore, modern agriculture is adopting new biofortification technologies to combat micronutrient deficiencies and improve agricultural productivity and sustainability. Biofertilization and nanofertilization practices are increasingly used due to their efficiency, safety, and reduced environmental impact. Biofertilizers are preparations of PGP-microorganisms that promote plant growth by influencing plant metabolism and improving the nutrient uptake, and nanofertilizers consist of synthesized nanoparticles with unique physicochemical properties that are capable of increasing plant nutrition and enriching agricultural products. This review presents the current micronutrient deficiencies associated with obesity, the modern unsustainable agri-food system contributing to obesity progression, and the development of bio- and nanofertilizers capable of biofortifying agri-food crops with micronutrients commonly deficient in patients with obesity.

Endophytic fungi: a tool for plant growth promotion and sustainable agriculture.

Endophytic fungi are found in most, if not all, plant species on the planet. They colonise inner plant tissues without causing symptoms of disease, thus providing benefits to the host plant while also benefiting from this interaction. The global concern for the development of more sustainable agriculture has increased in recent years, and research has been performed to decipher ecology and explore the potential of endophytic interactions in plant growth. To date, many studies point to the positive aspects of endophytic colonisation, and in this review, such research is summarised based on the direct (acquisition of nutrients and phytohormone production) and indirect (induced resistance, production of antibiotics and secondary metabolites, production of siderophores and protection for abiotic and biotic stresses) benefits of endophytic colonisation. An in-depth discussion of the mechanisms is also presented.

Efecto del riego con agua residual tratada sobre la calidad microbiológica del suelo y pasto King Grass / Effect of irrigation with treated wastewater on microbiological quality of the soil and King Grass

RESUMEN La contaminación del agua utilizada en el mundo y la falta de plantas de tratamiento altamente eficientes plantean problemas potenciales para la salud pública y el medio ambiente, por lo que los países buscan métodos confiables, inocuos y eficaces en función de los costos, para depurar las aguas residuales. Por otro lado, el sector agropecuario ha tomado auge, dada la necesidad de implementación de técnicas limpias, que sean amigables con el medio ambiente. El riego con aguas residuales en la agricultura, se ha vuelto común en regiones áridas y semiáridas, debido a la baja disponibilidad de agua, pero su uso requiere de un monitoreo cuidadoso de parámetros de higiene. El objetivo de este trabajo fue analizar el efecto del riego, con aguas residuales tratadas, sobre la contaminación microbiológica del suelo y el pasto King Grass, además de la producción de materia seca del forraje. Se evaluaron tres tratamientos: tratamiento T1: sin riesgo (SR), T2: agua subterránea (AS) y T3: agua residual tratada (ART). Se hicieron análisis fisicoquímicos y microbiológicos al agua, suelo y pasto. La contaminación microbiológica en el suelo regado con agua residual tratada no presentó diferencias significativas (p>0,05) frente a los suelos donde se aplicó riego con agua subterránea, así como tampoco se presentaron diferencias estadísticas significativas entre variables microbiológicas en pasto. Se concluyó, que la fuente de contaminación microbiológica de los pastos y el suelo no está relacionada únicamente con la carga microbiana presente en las aguas residuales, sino que se puede deber, entre otros factores, a las escorrentías y al uso de aguas subterráneas contaminadas para el riego.

ABSTRACT World water contamination and the lack of wastewater treatment plants pose potential problems concerning public health and on the environment. Therefore, countries are looking for reliable, safe and cost-effective methods to purify wastewater. On the other hand, the agricultural sector has taken a lot of momentum given the need to implement clean techniques that are friendly to the environment. boomed given the need to implement clean techniques that are friendly to the environment. Wastewater irrigation in agriculture has become common in arid and semi-arid regions due to low water availability, but its use requires careful monitoring of hygiene parameters. The objective of this work was to analyze the effect of treated wastewater on microbiological contamination of the soil and King grass, in addition to the production of dry matter from forage. Were evaluated three treatments: T1: no risk; T2: groundwater and T3: treated wastewater. Were made physicochemical and microbiological analyzes out on water, soil and grass. The microbiological contamination in the soil irrigated with treated wastewater did not show significant differences compared to the soils where irrigation with groundwater was applied, as well as there were no statistical differences between microbiological variables in grass. It was concluded that the source of microbiological contamination of pastures and soil is not only related to the microbial load present in the wastewater, but it may be due, among other factors, to runoff and the use of contaminated groundwater for irrigation.

Restoring degraded microbiome function with self-assembled communities.

The natural microbial functions of many soils are severely degraded. Current state-of-the-art technology to restore these functions is through the isolation, screening, formulation and application of microbial inoculants and synthetic consortia. These approaches have inconsistent success, in part due to the incompatibility between the biofertilizer, crop, climate, existing soil microbiome and physicochemical characteristics of the soils. Here, we review the current state of the art in biofertilization and identify two key deficiencies in current strategies: the difficulty in designing complex multispecies biofertilizers and the bottleneck in scaling the production of complex multispecies biofertilizers. To address the challenge of producing scalable, multispecies biofertilizers, we propose to merge ecological theory with bioprocess engineering to produce 'self-assembled communities' enriched for particular functional guilds and adapted to a target soil and host plant. Using the nitrogen problem as an anchor, we review relevant ecology (microbial, plant and environmental), as well as reactor design strategies and operational parameters for the production of functionally enriched self-assembled communities. The use of self-assembled communities for biofertilization addresses two major hurdles in microbiome engineering: the importance of enriching microbes indigenous to (and targeted for) a specific environment and the recognized potential benefits of microbial consortia over isolates (e.g. functional redundancy). The proposed community enrichment model could also be instrumental for other microbial functions such as phosphorus solubilization, plant growth promotion or disease suppression.

Exopolysaccharide production in Ensifer meliloti laboratory and native strains and their effects on alfalfa inoculation.

Bacterial surface molecules have an important role in the rhizobia-legume symbiosis. Ensifer meliloti (previously, Sinorhizobium meliloti), a symbiotic Gram-negative rhizobacterium, produces two different exopolysaccharides (EPSs), termed EPS I (succinoglycan) and EPS II (galactoglucan), with different functions in the symbiotic process. Accordingly, we undertook a study comparing the potential differences in alfalfa nodulation by E. meliloti strains with differences in their EPSs production. Strains recommended for inoculation as well as laboratory strains and native strains isolated from alfalfa fields were investigated. This study concentrated on EPS-II production, which results in mucoid colonies that are dependent on the presence of an intact expR gene. The results revealed that although the studied strains exhibited different phenotypes, the differences did not affect alfalfa nodulation itself. However, subtle changes in timing and efficacy to the effects of inoculation with the different strains may result because of other as-yet unknown factors. Thus, additional research is needed to determine the most effective inoculant strains and the best conditions for improving alfalfa production under agricultural conditions.

Comparative genomics of Paraburkholderia kururiensis and its potential in bioremediation, biofertilization, and biocontrol of plant pathogens.

Burkholderia harbors versatile Gram-negative species and is ß-Proteobacteria. Recently, it was proposed to split the genus in two main branches: one of animal and plant pathogens and another, Paraburkholderia, harboring environmental and plant-beneficial species. Currently, Paraburkholderia comprises more than 70 species with ability to occupy very diverse environmental niches. Herein, we sequenced and analyzed the genome of Paraburkholderia kururiensis type strain KP23T , and compared to P. kururiensis M130, isolated in Brazil, and P. kururiensis susbp. thiooxydans, from Korea. This study focused on the gene content of the three genomes with special emphasis on their potential of plant-association, biocontrol, and bioremediation. The comparative analyses revealed several genes related to plant benefits, including biosynthesis of IAA, ACC deaminase, multiple efflux pumps, dioxygenases, and degradation of aromatic compounds. Importantly, a range of genes for protein secretion systems (type III, IV, V, and VI) were characterized, potentially involved in P. kururiensis well documented ability to establish endophytic association with plants. These findings shed light onto bacteria-plant interaction mechanisms at molecular level, adding novel information that supports their potential application in bioremediation, biofertilization, and biocontrol of plant pathogens. P. kururiensis emerges as a promising model to investigate adaptation mechanisms in different ecological niches.

Biotechnological potential of plant growth-promoting bacteria from the roots and rhizospheres of endemic plants in ironstone vegetation in southeastern Brazil.

Microorganisms associated with plants have a great biotechnological potential, but investigations of these microorganisms associated with native plants in peculiar environments has been incipient. The objective of this study was to analyze the plant growth-promoting bacteria potential of cultivable bacteria associated with rare plants from the ferruginous rocky fields of the Brazilian Iron Quadrangle. The roots and rhizospheres of nine endemic plants species and samples of a root found in a lateritiric duricrust (canga) cave were collected, the culturable bacteria isolated and prospected for distinct biotechnological and ecological potentials. Out of the 148 isolates obtained, 8 (5.4%) showed potential to promote plant growth, whereas 4 (2.7%) isolates acted as biocontrol agents against Xanthomonas citri pathotype A (Xac306), reducing the cancrotic lesions by more than 60% when co-inoculated with this phytopathogen in Citrus sinensis plants. Moreover, other 4 (2.7%) isolates were classified as potential bioremediation agents, being able to withstand high concentrations of arsenite (5 mM As3+) and arsenate (800 mM As5+), by removing up to 35% and 15% of this metalloid in solution, respectively. These same four isolates had a positive influence on the growth of both the roots and the aerial parts when inoculated with tomato seeds in the soil contaminated with arsenic. This is the first time that an investigation highlights the potentialities of bacteria associated with rare plants of ferruginous rocky fields as a reservoir of microbiota of biotechnological and ecological interest, highlighting the importance of conservation of this area that is undergoing intense anthropic activity.

Benefits Associated with the Interaction of Endophytic Bacteria and Plants

ABSTRACT The endophytic bacteria belong to a larger group of microorganisms that have their life-cycle partly or entirely inside the plant and are located in intra and inter-cellular spaces or in the vascular tissue. These bacteria can be found colonizing aerial parts or roots. This review aims to analyze the colonization strategies of endophytic bacteria through interaction with plants, as well as to highlight the metabolic influence of these organisms in plant tissues, which result in physiological and biochemical changes. Depending on the different mechanisms used internally to colonize a plant, these microorganisms are called obligate, facultative, or passive endophytes. Phytostimulation, biofertilization and biological control are mechanisms that result in the development of the plant through the production of plant hormones, bioavailability of nutrients and antagonistic action to phytopathogens, respectively. The association between endophytic bacteria and plants features important benefits such as significant increases in growth, plant biomass, length of roots, dry matter production, and grain yield. Studies show that there is a great diversity of endophytic bacteria colonizing plant structures that result in several benefits to the host plant.

Efecto de inoculantes bacterianos edáficos mixtos en el desarrollo temprano de cultivares mejorados de cacao (Theobroma cacao L.) en un sistema agroforestal tradicional del norte de Oaxaca, México / Effect of mixed edaphic bacterial inoculants in the early development of improved cocoa cultivars (Theobroma cacao L.) in a traditional agroforestry system of Oaxaca, Mexico

El cacao (Theobroma cacao L.) es nativo de América del Sur y representa uno de los recursos «bioculturales¼ más significativos de Mesoamérica, ya que es una región donde se domesticó y tuvo relevancia como bebida ritual y como moneda en muchas culturas prehispánicas hasta la llegada de los españoles, quienes difundieron su uso en el mundo y lo convirtieron en una de las mercancías «commodity¼ más consumidas. Mediante este trabajo se propone una alternativa para atender la problemática de las plantaciones a través de la introducción de una diversidad amplia de cultivares de cacao en sistemas agroforestales tradicionales, en sinergia con la inoculación de bacterias edáficas fijadoras de nitrógeno y solubilizadoras de fósforo insoluble. En una parcela agroforestal tradicional se introdujeron plantas de cacao injertadas de cuatro cultivares y se dispusieron 3 tratamientos: aplicación de biofertilizante, aplicación de fertilizante químico y testigo. Se registraron la altura, el diámetro basal, el número de hojas y el número de ramas a los 2 y 12 meses, y se caracterizó la población de microorganismos asociada alrededor del tallo bajo la copa de las plantas. Los resultados de crecimiento muestran un buen potencial para los 4 cultivares estudiados y se observó que la biofertilización generó efectos significativos en algunos de los indicadores de crecimiento de las plantas de cacao. Así, las asociaciones vegetales en un sistema agroforestal podrían ser favorables para potenciar el desarrollo de frutos y resistencia a plagas y enfermedades.

Cocoa plant (Theobroma cacao L.) is native from South America and it represents one of the most significant "bio-cultural" resources of Mesoamerica, since it is a region where it was domesticated and had a relevance as ritual drink and as currency in many pre-hispanic cultures until the arrival of the Spaniards who spread its use worldwide, and became it one of the most consumed commodity goods. Through this research, an alternative is proposed to address the problem of cultivars through the introduction of a wide variety of cocoa plants in traditional agroforestry systems, in synergy with the inoculation of nitrogen-fixing and insoluble phosphor solubilizing edaphic bacterial consortia. Four cultivars of improved grafted cocoa plants were introduced in a traditional agroforestry plot and three fertilization treatments were applied: application of biofertilizer, application of chemical fertilizer and control. Measurements of height, stem diameter, number of leaves and branches were recorded at 2 and 12 months after planting and rhizosphere microbial populations were characterized. Growth results showed good potential for all studied cultivars and it was observed that biofertilization foresees significant effects in some of the growth indicators of cocoa plant. Thereby, plant associations in an agroforestry system could be favorable to promote fruit development and resistance to pests and diseases.

[Effect of mixed edaphic bacterial inoculants in the early development of improved cocoa cultivars (Theobroma cacao L.) in a traditional agroforestry system of Oaxaca, Mexico]. / Efecto de inoculantes bacterianos edáficos mixtos en el desarrollo temprano de cultivares mejorados de cacao (Theobroma cacao L.) en un sistema agroforestal tradicional del norte de Oaxaca, México.

Cocoa plant (Theobroma cacao L.) is native from South America and it represents one of the most significant "bio-cultural" resources of Mesoamerica, since it is a region where it was domesticated and had a relevance as ritual drink and as currency in many pre-hispanic cultures until the arrival of the Spaniards who spread its use worldwide, and became it one of the most consumed commodity goods. Through this research, an alternative is proposed to address the problem of cultivars through the introduction of a wide variety of cocoa plants in traditional agroforestry systems, in synergy with the inoculation of nitrogen-fixing and insoluble phosphor solubilizing edaphic bacterial consortia. Four cultivars of improved grafted cocoa plants were introduced in a traditional agroforestry plot and three fertilization treatments were applied: application of biofertilizer, application of chemical fertilizer and control. Measurements of height, stem diameter, number of leaves and branches were recorded at 2 and 12 months after planting and rhizosphere microbial populations were characterized. Growth results showed good potential for all studied cultivars and it was observed that biofertilization foresees significant effects in some of the growth indicators of cocoa plant. Thereby, plant associations in an agroforestry system could be favorable to promote fruit development and resistance to pests and diseases.

Bacillus spp: una alternativa para la promoción vegetal por dos caminos enzimáticos / Bacillus spp: An alternative for plant promotion by two enzymatic pathways

Resumen Objetivo. Se realizó una revisión sobre las características de las fitasas y nitrogenasas de Bacillus spp. y sus opciones de uso como alternativa biofertilizante. El género Bacillus es secretor de proteínas y metabolitos eficientes para el control de plagas y enfermedades, promueve el crecimiento vegetal a través de la solubilización de fósforo y la producción de reguladores de crecimiento como el ácido indol acético; así mismo participa en la fijación de nitrógeno cuando hace parte de consorcios microbianos. Como biofertilizante es una opción amigable para el suelo y el ambiente que da respuesta a la necesidad de implementar la agricultura sostenible.

Abstract Objective. We conducted a review of the characteristics of the phytase and nitrogenase of Bacillus sp. and their potential use as an alternative biological fertilizer. The genus Bacillus is an efficient secretor of proteins and metabolites; to control pests and diseases, promote plant growth through the solubilization of phosphorus and production of growth regulators as acetic indole. Likewise it is involved in nitrogen fixation when it is part of microbial consortia. As biological fertilizer is a friendly option for the soil and the environment that responds to the need to implement sustainable agriculture.

Different Behavior of Enteric Bacteria and Viruses in Clay and Sandy Soils after Biofertilization with Swine Digestate.

Enteric pathogens from biofertilizer can accumulate in the soil, subsequently contaminating water and crops. We evaluated the survival, percolation and leaching of model enteric pathogens in clay and sandy soils after biofertilization with swine digestate: PhiX-174, mengovirus (vMC0), Salmonella enterica Typhimurium and Escherichia coli O157:H7 were used as biomarkers. The survival of vMC0 and PhiX-174 in clay soil was significantly lower than in sandy soil (iT90 values of 10.520 ± 0.600 vs. 21.270 ± 1.100 and 12.040 ± 0.010 vs. 43.470 ± 1.300, respectively) and PhiX-174 showed faster percolation and leaching in sandy soil than clay soil (iT90 values of 0.46 and 2.43, respectively). S. enterica Typhimurium was percolated and inactivated more slowly than E. coli O157:H7 (iT90 values of 9.340 ± 0.200 vs. 6.620 ± 0.500 and 11.900 ± 0.900 vs. 10.750 ± 0.900 in clay and sandy soils, respectively), such that E. coli O157:H7 was transferred more quickly to the deeper layers of both soils evaluated (percolation). Our findings suggest that E. coli O157:H7 may serve as a useful microbial biomarker of depth contamination and leaching in clay and sandy soil and that bacteriophage could be used as an indicator of enteric pathogen persistence. Our study contributes to development of predictive models for enteric pathogen behavior in soils, and for potential water and food contamination associated with biofertilization, useful for risk management and mitigation in swine digestate recycling.

Formação de mudas de Nim com aplicação de biofertilizante bovino submetido à drenagem e estresse salino / Formation of Neem seedlings with application of bovine biofertilizer subjected to drainage and saline stress

Diversos fatores podem influenciar a formação de mudas. Entre as causas de insucesso, encontram-se a dificuldade de manutenção de drenagem e qualidade da água de irrigação. O objetivo do trabalho foi avaliar os efeitos do biofertilizante bovino e salinidade da água de irrigação na formação de mudas de nim. O delineamento experimental foi em blocos casualizados, com os tratamentos arranjados no esquema fatorial 5 x 2 x 2, com quatro repetições, referente aos níveis de salinidade da água de irrigação de 0,5; 1,5; 3,0; 4,5 e 6,0 dS m-1, no solo sem e com biofertilizante bovino, sem e com drenagem. As variáveis analisadas foram emergência de plântulas, número de folhas, diâmetro caulinar e área foliar. O biofertilizante bovino depois de diluído em água não salina (0,49 dS m-1) e não clorada na razão de 1:1, foi aplicado uma única vez, dois dias antes da semeadura, ao nível de 10% do volume do substrato. A irrigação das plantas com cada tipo de água foi feita diariamente fornecendo um volume de água suficiente para elevar a umidade do solo para o nível mais próxima da capacidade de campo. Pelos resultados, com o aumento da salinidade das águas houve reduções no crescimento das plantas em até 28%, mas o biofertilizante bovino e a drenagem do solo proporcionaram melhores condições de crescimento das mudas.

Several factors may influence in formation of seedlings. Among the causes of failure find discovered the difficulty of maintain of the drainage and quality of irrigation water. The objective of wok was study or evaluate the effects of bovine biofertilizer and water salinity in the formation of neem seedlings. The experimental design was in randomized block, with the treatments arranged in factorial scheme 5 x 2 x 2, with four repetitions, referring at the five salinity levels of irrigation water: 0.5, 1.5, 3.0, 4.5 and 6.0 dS m-1, in soil without and with bovine biofertilizer, in soil without and with soil drainage. The variables analyzed were: seedlings emergence, number of leaves, stem diameter and leaf area. The bovine biofertilizer after diluted in noon-saline water (0,49 dS m-1) e not chlorinated in reason to 1:1, was applied an once two days before sowing, at level to 10% of substrate volume. The plants were irrigated daily with each type of water in volume sufficient to increase the soil moisture for the level nearest of field capacity. By results, with the increasing of salinity waters there were reductions in plant growth up to 28%, but the bovine biofertilizer and soil drainage provided better conditions for seedling growth.