A "love match" score to compare root exudate attraction and feeding of the plant growth-promoting rhizobacteria Bacillus subtilis, Pseudomonas fluorescens, and Azospirillum brasilense.

Introduction: The rhizosphere is the zone of soil surrounding plant roots that is directly influenced by root exudates released by the plant, which select soil microorganisms. The resulting rhizosphere microbiota plays a key role in plant health and development by enhancing its nutrition or immune response and protecting it from biotic or abiotic stresses. In particular, plant growth-promoting rhizobacteria (PGPR) are beneficial members of this microbiota that represent a great hope for agroecology, since they could be used as bioinoculants for sustainable crop production. Therefore, it is necessary to decipher the molecular dialog between roots and PGPR in order to promote the establishment of bioinoculants in the rhizosphere, which is required for their beneficial functions. Methods: Here, the ability of root exudates from rapeseed (Brassica napus), pea (Pisum sativum), and ryegrass (Lolium perenne) to attract and feed three PGPR (Bacillus subtilis, Pseudomonas fluorescens, and Azospirillum brasilense) was measured and compared, as these responses are directly involved in the establishment of the rhizosphere microbiota. Results: Our results showed that root exudates differentially attracted and fed the three PGPR. For all beneficial bacteria, rapeseed exudates were the most attractive and induced the fastest growth, while pea exudates allowed the highest biomass production. The performance of ryegrass exudates was generally lower, and variable responses were observed between bacteria. In addition, P. fluorescens and A. brasilense appeared to respond more efficiently to root exudates than B. subtilis. Finally, we proposed to evaluate the compatibility of each plant-PGPR couple by assigning them a "love match" score, which reflects the ability of root exudates to enhance bacterial rhizocompetence. Discussion: Taken together, our results provide new insights into the specific selection of PGPR by the plant through their root exudates and may help to select the most effective exudates to promote bioinoculant establishment in the rhizosphere.

Effect of a Bacillus velezensis and Lysinibacillus fusiformis-based biofertilizer on phosphorus acquisition and grain yield of soybean.

Introduction: Phosphate-solubilizing bacteria that function through acidification (organic acid synthesis) or mineralization (production of enzymes such as phytase and phosphatases) have been explored as a biotechnological alternative to enhance plant access to phosphorus (P) retained in organic and inorganic forms in agricultural soils. This study tested the hypothesis that applying a biofertilizer composed of a recognized phosphate-solubilizing bacterium (Bacillus velezensis - endophytic strain BVPS01) and an underexplored plant growth-promoting bacterium (Lysinibacillus fusiformis - endophytic strain BVPS02) would improve the growth and grain yield of Glycine max L. plants. Methods: Initial in vitro tests assessed the functional traits of these bacteria, and a mix of strains BVPS01 and BVPS02 was produced and tested under field conditions to evaluate its agronomic efficiency. Results: The results confirmed the hypothesis that the tested biofertilizer enhances the agronomic performance of G. max plants in the field. The B. velezensis strain (BVPS01) was found to be more effective than the L. fusiformis strain (BVPS02) in solubilizing phosphates via the phosphatase enzyme production pathway, indicated by the expression of the phoC and phoD genes. In contrast, L. fusiformis was more effective in solubilizing phosphates through organic acid and phytase-related pathways, in addition to synthesizing indole-3-acetic acid and increasing the mitotic index in the root meristem of G. max plants. These strains exhibited biological compatibility, and the formulated product based on these rhizobacteria enhanced root development and increased the number of nodules and flowers, positively affecting 1000-grain weight, grain yield, and grain P content. Discussion: Thus, the tested biofertilizer demonstrated potential to improve root growth and increase both the yield and quality of soybean crops, making it a sustainable and low-cost strategy.

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.

Tracking maize colonization and growth promotion by Azospirillum reveals strain-specific behavior and the influence of inoculation method.

Inoculation of Azospirillum in maize has become a standard practice in Latin America. However, information on the behavior and population survival of the Azospirillum post-inoculation is scarce, making standardization difficult and generating variations in inoculation efficiency across assays. In this study, we tracked the colonization of three agriculturally relevant Azospirillum strains (Ab-V5, Az39, and the ammonium excreting HM053) after different inoculation methods in maize crops by qPCR. Besides, we assessed their ability to promote maize growth by measuring biometric parameters after conducting a greenhouse essay over 42 days. Inoculated plants exhibited Azospirillum population ranging from 103 to 107 cells plant-1 throughout the experiment. While all strains efficiently colonized roots, only A. argentinense Az39 demonstrated bidirectional translocation between roots and shoots, which characterizes a systemic behavior. Optimal inoculation methods for plant growth promotion varied among strains: soil inoculation promoted the best maize growth for the Ab-V5 and Az39 strains, while seed inoculation proved most effective for HM053. The findings of this study demonstrate that the inoculation method affects the behavior of Azospirillum strains and their effectiveness in promoting maize growth, thereby guiding practices to enhance crop yield.

Diversity of bacteria of the genus Sphingomonas associated with sugarcane (Saccharum spp.) culm apoplast fluid and their agrotechnological potential.

In sugarcane, sequences related to the genus Sphingomonas have been widely detected by microbiome studies. In this work, the presence of bacteria of this genus was confirmed using culture-dependent and independent techniques. A collection of thirty isolates was obtained using semispecific cultivation conditions, and a specific PCR assay was applied to help confirm the isolates as belonging to the genus. A series of laboratory evaluations were carried out to identify potential properties among the isolates in the collection, which consequently allowed the identification of some most promising isolates for the development of new agricultural bioinputs. In a separate analysis, the culture-independent fluorescence in situ hybridization (FISH) methodology was applied to demonstrate the natural occurrence of Sphingomonas in different organs and tissues of sugarcane. The results showed the presence of bacteria of the genus in the spaces between cells (apoplast) of the culm parenchyma, in vessels in the region of the leaf vein, on the adaxial surface of the leaf blade, and on the root surface, sometimes close to the base of root hairs, which suggests extensive colonization on the host plant. In summary, the present study corroborates previous metagenomic amplicon sequencing results that indicated a high occurrence of Sphingomonas associated with sugarcane. This is the first study that uses approaches other than amplicon sequencing to confirm the occurrence of the genus in sugarcane and, at the same time, demonstrates potentially beneficial activities to be explored by sugarcane cultivation.

Isolation and Characterization of Plant Growth-Promoting Bacteria from the Rhizosphere of Medicinal and Aromatic Plant Minthostachys verticillata.

This study aimed to isolate and characterize Pseudomonas native strains from the rhizospheric soil of Minthostachys verticillata plants to evaluate their potential as plant growth-promoting rhizobacteria (PGPR). A total of 22 bacterial isolates were obtained and subjected to various biochemical tests, as well as assessments of plant growth-promoting traits such as phosphate solubilization, hydrogen cyanide production, biocontrol properties through antibiosis, and indole acetic production. Genotypic analysis via 16S rRNA gene sequencing and phylogenetic tree construction identified the strains, with one particular strain named SM 33 showing significant growth-promoting effects on M. verticillata seedlings. This strain, SM 33, showed high similarity to Stutzerimonas stutzeri based on 16S rRNA gene sequencing and notably increased both shoot fresh weight and root dry weight of the plants. These findings underscore the potential application of native Pseudomonas strains in enhancing plant growth and health, offering promising avenues for sustainable agricultural practices.

The Biotechnological Potential of Plant Growth-Promoting Rhizobacteria Isolated from Maize (Zea mays L.) Cultivations in the San Martin Region, Peru.

Maize (Zea mays L.) is an essential commodity for global food security and the agricultural economy, particularly in regions such as San Martin, Peru. This study investigated the plant growth-promoting characteristics of native rhizobacteria isolated from maize crops in the San Martin region of Peru with the aim of identifying microorganisms with biotechnological potential. Soil and root samples were collected from maize plants in four productive zones in the region: Lamas, El Dorado, Picota, and Bellavista. The potential of twelve bacterial isolates was evaluated through traits, such as biological nitrogen fixation, indole acetic acid (IAA) production, phosphate solubilization, and siderophore production, and a completely randomized design was used for these assays. A completely randomized block design was employed to assess the effects of bacterial strains and nitrogen doses on maize seedlings. The B3, B5, and NSM3 strains, as well as maize seeds of the yellow hard 'Advanta 9139' variety, were used in this experiment. Two of these isolates, B5 and NSM3, exhibited outstanding characteristics as plant growth promoters; these strains were capable of nitrogen fixation, IAA production (35.65 and 26.94 µg mL-1, respectively), phosphate solubilization (233.91 and 193.31 µg mL-1, respectively), and siderophore production (34.05 and 89.19%, respectively). Furthermore, molecular sequencing identified the NSM3 isolate as belonging to Sporosarcina sp. NSM3 OP861656, while the B5 isolate was identified as Peribacillus sp. B5 OP861655. These strains show promising potential for future use as biofertilizers, which could promote more sustainable agricultural practices in the region.

Plant growth-promoting abilities of Methylobacterium sp. 2A involve auxin-mediated regulation of the root architecture.

Methylobacterium sp. 2A, a plant growth-promoting rhizobacteria (PGPR) able to produce indole-3-acetic acid (IAA), significantly promoted the growth of Arabidopsis thaliana plants in vitro. We aimed to understand the determinants of Methylobacterium sp. 2A-A. thaliana interaction, the factors underlying plant growth-promotion and the host range. Methylobacterium sp. 2A displayed chemotaxis to methanol and formaldehyde and was able to utilise 1-aminocyclopropane carboxylate as a nitrogen source. Confocal microscopy confirmed that fluorescent protein-labelled Methylobacterium sp. 2A colonises the apoplast of A. thaliana primary root cells and its inoculation increased jasmonic and salicylic acid in A. thaliana, while IAA levels remained constant. However, inoculation increased DR5 promoter activity in root tips of A. thaliana and tomato plants. Inoculation of this PGPR partially restored the agravitropic response in yucQ mutants and lateral root density was enhanced in iaa19, arf7, and arf19 mutant seedlings. Furthermore, Methylobacterium sp. 2A volatile organic compounds (VOCs) had a dose-dependent effect on the growth of A. thaliana. This PGPR is also able to interact with monocots eliciting positive responses upon inoculation. Methylobacterium sp. 2A plant growth-promoting effects can be achieved through the regulation of plant hormone levels and the emission of VOCs that act either locally or at a distance.

Cost Reduction in the Production of Green Dwarf Coconut Palm Seedlings Biostimulated with Bacillus cereus.

The production of coconut tree seedlings is an important step in the production process, as it substantially affects the productive performance of the adult plant, and the way of obtaining seedlings directly reflects the added costs. To minimize costs, the introduction of biostimulants can be considered a viable and sustainable technology. This study aimed to evaluate the effects of applying Bacillus cereus in promoting growth and reducing the costs of producing Brazilgreen dwarf coconut seedlings. The study has two stages, the first was an experiment carried out in a commercial nursery in the state of Pará-Brazil. The design was completely randomized, with two treatments: control with water (100% mineral fertilization) and B. cereus inoculation (50% mineral fertilization), with 10 replicates each. Biometric parameters and the quality of seedlings were evaluated. In the second stage, the production of stimulated seedlings was compared to that of commercial seedlings, and the effective operating cost (COE) and the total operating cost (TOC) were determined. Biostimulation with B. cereus promotes the growth of coconut tree seedlings, increases seedling quality, and reduces nursery time. In addition, the cost of production is reduced by 10%. Thus, microbial technology is a positive strategy for the production of Brazilian green dwarf coconut seedlings. Using B. cereus can guarantee obtaining seedlings with high performance and at a lower cost. These results may favor obtaining adult plants with high productivity. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-023-01163-9.

Synergistic enhancement of plant growth and cadmium stress defense by Azospirillum brasilense and plant heme: Modulating the growth-defense relationship.

Plant growth-promoting rhizobacteria (PGPR) play important roles in plant growth and defense under heavy metal (HM) stress. The direct integration of microbial and plant signals is key to the regulation of plant growth and HM stress defense, but the underlying mechanisms are still limited. Herein, we reveal a novel mechanism by which PGPR regulates plant growth-regulating substances in plant tissues and coordinates plant growth and defense in pak choi under cadmium (Cd) stress. This might be an efficient strategy and an extension of the mechanism by which plant-microbe interactions improve plant stress resistance. Azospirillum brasilense and heme synergistically reduced the shoot Cd content and promoted the growth of pak choi. The interaction between abscisic acid of microbial origin and heme improved Cd stress tolerance through enhancing Cd accumulation in the root cell wall. The interaction between A. brasilense and heme induced the growth-defense shift in plants under Cd stress. Plants sacrifice growth to enhance Cd stress defense, which then transforms into a dual promotion of both growth and defense. This study deepens our understanding of plant-microbe interactions and provides a novel strategy to improve plant growth and defense under HM stress, ensuring future food production and security.

Simultaneous Impact of Rhizobacteria Inoculation and Leaf-Chewing Insect Herbivory on Essential Oil Production and VOC Emissions in Ocimum basilicum.

Inoculation with rhizobacteria and feeding by herbivores, two types of abiotic stress, have been shown to increase the production of secondary metabolites in plants as part of the defense response. This study explored the simultaneous effects of inoculation with Bacillus amyloliquefaciens GB03 (a PGPR species) and herbivory by third-instar Spodoptera frugiperda larvae on essential oil (EO) yield and volatile organic compound (VOC) emissions in Ocimum basilicum plants. The density of glandular trichomes was also examined, given that they are linked to EO production and VOC emission. Herbivory increased EO content, but inoculation on its own did not. When combined, however, the two treatments led to a 10-fold rise in EO content with respect to non-inoculated plants. VOC emissions did not significantly differ between inoculated and non-inoculated plants, but they doubled in plants chewed by the larvae with respect to their undamaged counterparts. Interestingly, no changes were observed in VOC emissions when the treatments were tested together. In short, the two biotic stressors elicited differing plant defense responses, mainly when EO was concerned. PGPR did not stimulate EO production, while herbivory significantly enhanced it and increased VOC emissions. The combined treatment acted synergistically, and in this case, PGPR inoculation may have had a priming effect that amplified plant response to herbivory. Peltate trichome density was higher in inoculated plants, those damaged by larvae, and those subjected to the combination of both treatments. The findings highlight the intricate nature of plant defense mechanisms against various stressors and hint at a potential strategy to produce essential oil through the combined application of the two stressors tested here.

Plant seedlings of peas, tomatoes, and cucumbers exude compounds that are needed for growth and chemoattraction of Rhizobium leguminosarum bv. viciae 3841 and Azospirillum brasilense Sp7.

This study characterizes seedling exudates of peas, tomatoes, and cucumbers at the level of chemical composition and functionality. A plant experiment confirmed that Rhizobium leguminosarum bv. viciae 3841 enhanced growth of pea shoots, while Azospirillum brasilense Sp7 supported growth of pea, tomato, and cucumber roots. Chemical analysis of exudates after 1 day of seedling incubation in water yielded differences between the exudates of the three plants. Most remarkably, cucumber seedling exudate did not contain detectable sugars. All exudates contained amino acids, nucleobases/nucleosides, and organic acids, among other compounds. Cucumber seedling exudate contained reduced glutathione. Migration on semi solid agar plates containing individual exudate compounds as putative chemoattractants revealed that R. leguminosarum bv. viciae was more selective than A. brasilense, which migrated towards any of the compounds tested. Migration on semi solid agar plates containing 1:1 dilutions of seedling exudate was observed for each of the combinations of bacteria and exudates tested. Likewise, R. leguminosarum bv. viciae and A. brasilense grew on each of the three seedling exudates, though at varying growth rates. We conclude that the seedling exudates of peas, tomatoes, and cucumbers contain everything that is needed for their symbiotic bacteria to migrate and grow on.

The elite strain INPA03-11B approved as a cowpea inoculant in Brazil represents a new Bradyrhizobium species and it has high adaptability to stressful soil conditions.

The strain INPA03-11BT, isolated in the 1980s from nodules of Centrosema sp. collected in Manaus, Amazonas, Brazil, was approved by the Brazilian Ministry of Agriculture as a cowpea inoculant in 2004. Since then, several studies have been conducted regarding its phenotypic, genetic, and symbiotic characteristics under axenic and field conditions. Phenotypic features demonstrate its high adaptability to stressful soil conditions, such as tolerance to acidity, high temperatures, and 13 antibiotics, and, especially, its high symbiotic efficiency with cowpea and soybean, proven in the field. The nodC and nifH phylogenies placed the INPA strain in the same clade as the species B. macuxiense BR 10303T which was also isolated from the Amazon region. The sequencing of the 16S rRNA ribosomal gene and housekeeping genes, as well as BOX-PCR profiles, showed its potential as a new species, which was confirmed by a similarity percentage of 94.7% and 92.6% in Average Nucleotide Identity with the closest phylogenetically related species Bradyrhizobium tropiciagri CNPSo1112T and B. viridifuturi SEMIA690T, respectively. dDDH values between INPA03-11BT and both CNPSo 1112T and SEMIA690T were respectively 58.5% and 48.1%, which are much lower than the limit for species boundary (70%). Therefore, we propose the name Bradyrhizobium amazonense for INPA03-11BT (= BR3301 = SEMIA6463).

Whole genome analysis of Bacillus velezensis 160, biological control agent of corn head smut.

Corn head smut is a disease caused by the fungus Sporisorium reilianum. This phytosanitary problem has existed for several decades in the Mezquital Valley, an important corn-producing area in central Mexico. To combat the problem, a strain identified as Bacillus subtilis 160 was applied in the field, where it decreased disease incidence and increased crop productivity. In this study, the sequencing and analysis of the whole genome sequence of this strain were carried out to identify its genetic determinants for the production of antimicrobials. The B. subtilis 160 strain was found to be Bacillus velezensis. Its genome has a size of 4,297,348 bp, a GC content of 45.8%, and 4,174 coding sequences. Comparative analysis with the genomes of four other B. velezensis strains showed that they share 2,804 genes and clusters for the production of difficidin, bacillibactin, bacilysin, macrolantin, bacillaene, fengycin, butirosin A, locillomycin, and surfactin. For the latter metabolite, unlike the other strains that have only one cluster, B. velezensis 160 has three. A cluster for synthesizing laterocidine, an antimicrobial reported only in Brevibacillus laterosporus, was also identified. IMPORTANCE: In this study, we performed sequencing and analysis of the complete genome of the strain initially identified as Bacillus subtilis 160 as part of its characterization. This bacterium has shown its ability to control corn head smut in the field, a disease caused by the basidiomycete fungus Sporisorium reilianum. Analyzing the complete genome sequence not only provides a more precise taxonomic identification but also sheds light on the genetic potential of this bacterium, especially regarding mechanisms that allow it to exert biological control. Employing molecular and bioinformatics tools in studying the genomes of agriculturally significant microorganisms offers insights into the development of biofungicides and bioinoculants. These innovations aim to enhance plant growth and pave the way for strategies that boost crop productivity.

Nitric oxide synthase expression in Pseudomonas koreensis MME3 improves plant growth promotion traits.

The development of novel biotechnologies that promote a better use of N to optimize crop yield is a central goal for sustainable agriculture. Phytostimulation, biofertilization, and bioprotection through the use of bio-inputs are promising technologies for this purpose. In this study, the plant growth-promoting rhizobacteria Pseudomonas koreensis MME3 was genetically modified to express a nitric oxide synthase of Synechococcus SyNOS, an atypical enzyme with a globin domain that converts nitric oxide to nitrate. A cassette for constitutive expression of synos was introduced as a single insertion into the genome of P. koreensis MME3 using a miniTn7 system. The resulting recombinant strain MME3:SyNOS showed improved growth, motility, and biofilm formation. The impact of MME3:SyNOS inoculation on Brachypodium distachyon growth and N uptake and use efficiencies under different N availability situations was analyzed, in comparison to the control strain MME3:c. After 35 days of inoculation, plants treated with MME3:SyNOS had a higher root dry weight, both under semi-hydroponic and greenhouse conditions. At harvest, both MME3:SyNOS and MME3:c increased N uptake and use efficiency of plants grown under low N soil. Our results indicate that synos expression is a valid strategy to boost the phytostimulatory capacity of plant-associated bacteria and improve the adaptability of plants to N deficiency. KEY POINTS: • synos expression improves P. koreensis MME3 traits important for rhizospheric colonization • B. distachyon inoculated with MME3:SyNOS shows improved root growth • MME3 inoculation improves plant N uptake and use efficiencies in N-deficient soil.

Isolation and identification of a new Bacillus glycinifermentans strain from date palm rhizosphere and its effect on barley seeds under heavy metal stress.

Soil contamination by heavy metals is one of the major problems that adversely decrease plant growth and biomass production. Inoculation with the plant growth-promoting rhizobacteria (PGPR) can attenuate the toxicity of heavy metals and enhancing the plant growth. In this study, we evaluated the potential of a novel extremotolerant strain (IS-2 T) isolated from date palm rhizosphere to improve barley seedling growth under heavy metal stress. The species-level identification was carried out using morphological and biochemical methods combined with whole genome sequencing. The bacterial strain was then used in vitro for inoculating Hordeum vulgare L. exposed to three different Cr, Zn, and Ni concentrations (0.5, 1, and 2 mM) in petri dishes and different morphological parameters were assessed. The strain was identified as Bacillus glycinifermentans species. This strain showed high tolerance to pH (6-11), salt stress (0.2-2 M), and heavy metals. Indeed, the minimum inhibitory concentrations at which bacterium was unable to grow were 4 mM for nickel, 3 mM for zinc, more than 8 mM for copper, and 40 mM for chromium, respectively. It was observed that inoculation of Hordeum vulgare L. under metal stress conditions with Bacillus glycinifermentans IS-2 T stain improved considerably the growth parameters. The capacity of the IS-2 T strain to withstand a range of abiotic stresses and improve barley seedling development under lab conditions makes it a promising candidate for use as a PGPR in zinc, nickel, copper, and chromium bioremediation.

Enhancing Water Status and Nutrient Uptake in Drought-Stressed Lettuce Plants (Lactuca sativa L.) via Inoculation with Different Bacillus spp. Isolated from the Atacama Desert.

Drought is a major challenge for agriculture worldwide, being one of the main causes of losses in plant production. Various studies reported that some soil's bacteria can improve plant tolerance to environmental stresses by the enhancement of water and nutrient uptake by plants. The Atacama Desert in Chile, the driest place on earth, harbors a largely unexplored microbial richness. This study aimed to evaluate the ability of various Bacillus sp. from the hyper arid Atacama Desert in the improvement in tolerance to drought stress in lettuce (Lactuca sativa L. var. capitata, cv. "Super Milanesa") plants. Seven strains of Bacillus spp. were isolated from the rhizosphere of the Chilean endemic plants Metharme lanata and Nolana jaffuelii, and then identified using the 16s rRNA gene. Indole acetic acid (IAA) production, phosphate solubilization, nitrogen fixation, and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity were assessed. Lettuce plants were inoculated with Bacillus spp. strains and subjected to two different irrigation conditions (95% and 45% of field capacity) and their biomass, net photosynthesis, relative water content, photosynthetic pigments, nitrogen and phosphorus uptake, oxidative damage, proline production, and phenolic compounds were evaluated. The results indicated that plants inoculated with B. atrophaeus, B. ginsengihumi, and B. tequilensis demonstrated the highest growth under drought conditions compared to non-inoculated plants. Treatments increased biomass production and were strongly associated with enhanced N-uptake, water status, chlorophyll content, and photosynthetic activity. Our results show that specific Bacillus species from the Atacama Desert enhance drought stress tolerance in lettuce plants by promoting several beneficial plant traits that facilitate water absorption and nutrient uptake, which support the use of this unexplored and unexploited natural resource as potent bioinoculants to improve plant production under increasing drought conditions.

Enterobacter sp. DBA51 produces ACC deaminase and promotes the growth of tomato (Solanum lycopersicum L.) and tobacco (Nicotiana tabacum L.) plants under greenhouse condition.

Bacterial isolated from rhizospheric soil associated with the semi-desertic plant Coronilla juncea L. were screened for 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity, a common trait for plant-growth-promoting rhizobacteria (PGPR). Among bacterial isolates, strain DBA51 showed phosphate solubilizing index (PSI), producing indole acetic acid (IAA), and with the hemolysis-negative test. Sequencing and analysis of the 16S rDNA gene identified DBA51 as Enterobacter. DBA51 did not show antagonistic activity in vitro against bacterial (Clavibacter michiganensis, Pseudomonas syringae pv. tomato DC3000 and Pectobacterium cacticidum FHLGJ22) and fungal phytopathogens (Alternaria sp., Fusarium oxysporum fsp. lycopersici, Fusarium oxysporum fsp. cubense M5, and Rhizoctonia sp.). Root inoculations with DBA51 in tomato (Solanum lycopersicum L.) and tobacco (Nicotiana tabacum L.) plants were performed under greenhouse conditions. Plant height (20 %) and root biomass (40 %) were significantly enhanced in tomato plants inoculated with DBA51 compared to non-inoculated plants, although for tobacco plants, only root biomass (27 %) showed significant differences with DBA51. In addition, physiological parameters such as photosynthetic rate (µmol CO2 m-2 s-1), stomatal conductance (mol H2O m-2 s-1), and transpiration rate (mmol H2O m-2 s-1) were also evaluated, and no differences were detected between DBA51-inoculated and control treatment in tomato and tobacco leaves. The observed results indicate that the DBA51 strain could be used as a biofertilizer to improve yields of horticultural crops.

Multi-trait efficiency and interactivity of bacterial consortia used to enhance plant performance under water stress conditions.

Water stress is a major limiting factor for agricultural production under current and projected climate change scenarios. As a sustainable strategy, plant growth-promoting bacterial consortia have been used to reduce plant water stress. However, few studies have examined the effects of stress on multi-trait efficiency and interactivity of bacterial species. In this study, we used several in-vitro experiments, plant assays and greenhouse trials to investigate the effects of stress and bacterial consortia on 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activities, indole-3-acetic acid (IAA) production and plant growth-promoting traits (Phosphate-solubilization, starch hydrolysis, siderophores and ammonium production). We further assessed biofilm formation and the chemotactic behaviour in response to ACC. A total of fifteen ACCD rhizobacteria with multiple growth-promoting traits from the dominant plant species from the hyperseasonal Aripo Savannas were screened in this study. Five of the isolates were further analyzed based on their ACCD activities and were tested in single and dual consortium to assess their abilities in promoting growth under simulated drought stress (-0.35 MPa) and chemically induced ACC conditions (0.03 mM). Our findings showed that bacteria which produce high concentrations of IAA affected the isolates' ability to promote growth under stress, irrespective of microbial combination with ACCD activity above the minimal threshold of 20 nmol α-ketobutyrate mg-1 h-1. Biofilm production with co-culture interaction varied greatly across treatments, however, the general trend showed an increase in biofilm under stress induce conditions. The best performing co-culture, UWIGT-83 and UWIGT-120 (Burkholderia sp.) showed enhanced growth in germination assays and in greenhouse trials with Capsicum chinense (Moruga red hot peppers) under drought stress, when compared to non-inoculated treatments. The findings highlight the importance of testing interactivity of bacterial species with multiple growth promoting traits under stress conditions; and proposed the use of ACC growth media as a novel biofilm screening method for selecting potential stress plant growth-promoting bacteria. Better screening strategies for appropriate plant growth-promoting bacteria may narrow the inconsistency observed between laboratory and field trials.

N- acyl homoserine lactones (AHLs) type signal molecules produced by rhizobacteria associated with plants that growing in a metal(oids) contaminated soil: A catalyst for plant growth.

The present study explores the potential of rhizobacteria isolated from Baccharis linearis and Solidago chilensis in metal(loid)-contaminated soil for producing N-acyl-homoserine lactones (AHLs)-type signal molecules and promoting plant growth. A total of 42 strains were isolated, four demonstrating the production of AHL-type signal molecules. Based on 16S rRNA gene sequencing analyses and MALDI-TOF analyses, these four isolates were identified as belonging to the Pseudomonas genus, specifically P. brassicacearum, P. frederickberguensis, P. koreensis, and P. orientalis. The four AHL-producing strains were evaluated for metal(loid)s tolerance, their plant growth promotion traits, AHL quantification, and their impact on in vitro Lactuca sativa plant growth. The study found that four strains exhibited high tolerance to metal(loid)s, particularly As, Cu, and Zn. Additionally, plant growth-promoting traits were detected in AHL-producing bacteria, such as siderophore production, ammonia production, ACC deaminase activity, and P solubilization. Notably, AHL production varied among strains isolated from B. linearis, where C7-HSL and C9-HSL signal molecules were detected, and S. chilensis, where only C7-HSL signal molecules were observed. In the presence of copper, the production of C7-HSL and C9-HSL significantly decreased in B. linearis isolates, while in S. chilensis isolates, C7-HSL production was inhibited. Further, when these strains were inoculated on lettuce seeds and in vitro plants, a significant increase in germination and plant growth was observed. Mainly, the inoculation of P. brassicacearum and P. frederickberguensis led to extensive root hair development, significantly increasing length and root dry weight. Our results demonstrate that rhizospheric strains produce AHL molecules and stimulate plant growth, primarily through root development. However, the presence of copper reduces the production of these molecules, potentially affecting the root development of non-metalloid tolerant plants such as S. chilensis, which would explain its low population in this hostile environment.