Search Results (14)

Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium capable of colonizing different host plants. This work evaluated the impact of Colombian native isolate G. diazotrophicus GIBI029 on tomato yield and quality in response to nitrogen and phosphorus fertilization levels. The experiment was conducted under semi-controlled conditions using a split-plot design with four replicates and ten plants per treatment. Variables assessed included fruit count per plant, fruit weight, average fruit weight, production per plant, and yield. Application of GIBI029 without fertilization resulted in a higher number and weight of fruits per plant across harvests (7.1 fruits, 509.2 g) compared to both the unfertilized control (4.8 fruits, 271.7 g) and with complete nitrogen and phosphorus fertilization (5.2 fruits, 288.8 g). The behavior of these variables were similar for GIBI029 and complete fertilization (7.0 fruits and 510.7 g per harvest). The highest yields were obtained with GIBI029 without fertilization (106.1 t ha⁻¹) and with full nitrogen and phosphorus fertilization (106.4 t ha⁻¹). Under the evaluated conditions, native G. diazotrophicus GIBI029 isolate could effectively improve tomato growth and yield in contrast to the controls. Based on these findings, the reproducibility of this behavior should be confirmed, and the mechanisms involved in the plant–bacteria interaction should be determined. Full article

Two novel marine hydrogen- and sulfur-oxidizing bacteria, designated HSL1-7^T and HSL3-1^T, were isolated from mangrove sediments from Fujian Province, China. Strain HSL1-7^T exhibited Gram-negative, rod-shaped to slightly curved morphology with polar flagellum-driven motility, whereas strain HSL3-1^T was Gram-negative, rod-shaped and non-motile. Strain HSL1-7^T and HSL3-1^T were obligate chemolithoautotrophs, capable of using molecular hydrogen and thiosulfate as an energy source, and molecular oxygen and elemental sulfur as the electron acceptors for growth. Cellular fatty acid profiles revealed similar predominant components (C_16:1ω7c, C_16:0, C_18:1ω7c, and C_14:0) in both strains. Strains HSL1-7^T and HSL3-1^T were strongly diazotrophic, as demonstrated by ¹⁵N₂ fixation when a fixed nitrogen source was absent from the growth medium. The DNA G+C contents of strains HSL1-7^T and HSL3-1^T were determined to be 36.1% and 57.3%, respectively. Based on the 16S rRNA gene sequences, strains HSL1-7^T and HSL3-1^T exhibited the highest sequence similarities with Sulfurimonas marina B2^T (98.5% and 94.45%, respectively). Notably, the 16S rRNA gene sequence similarity between strains HSL1-7^T and HSL3-1^T was 93.19%, indicating that they represent distinct species within the genus Sulfurimonas. Comparative genomic analyses revealed the presence of diverse metabolic profiles in strains HSL1-7^T and HSL3-1^T, including carbon fixation, hydrogen oxidation, sulfur oxidation, and nitrogen fixation. The combined phenotypic, chemotaxonomic, and phylogenetic evidence, including average nucleotide identity and in silico DNA-DNA hybridization values, shows that strains HSL1-7^T and HSL3-1^T represent two novel species of the genus Sulfurimonas for which the names Sulfurimonas microaerophilic sp. nov. and Sulfurimonas diazotrophicus sp. nov. are proposed, with the type strains HSL1-7^T (=MCCC 1A18899^T = KCTC 25640^T) and HSL3-1^T (=MCCC 1A18844^T), respectively. Full article

Plant biostimulants (PBs) have been considered the new wave for ecological intensification and sustainability, but are they sustainable? They increase nutrient use efficiency and reduce the impact of abiotic stress in plants. However, commercially available PBs based on humic substances are obtained using non-renewable sources of organic matter. At the same time, the microbial inoculants include a discussion of the properties of microorganisms and formulation design, as well as standards of purity and process control. Farmers depend on biological inputs like others to generate additional income for agribusiness. We produced a composite PB using humic substances isolated from vermicompost with KOH 5% and microbial consortia of plant growth-promoting bacteria (H. seropedicae, G. diazotrophicus, Bacillus spp.) grown in a simple medium with molasses and fishmeal as sources of C and N, respectively, in a homemade reactor at 37 °C for 36 h. The on-farm PB was applied directly in a passion fruit trial, and plant health and yield were monitored. The plants treated with the PB showed decreased visual symptoms of pests and diseases concurrent to higher activities of the enzymes used to monitor the induction of the plant resistance system (1,3-β glucanase, peroxidase, and phenylalanine ammonia-lyase). Plants treated with the PB yielded more than 50% more passion fruit than the control in soil with natural low fertility, fertilized with vermicompost. It is possible to produce PBs directly on the farm, leveraging locally available resources and simple technologies to sustainably enhance plant health and productivity. Full article

Bacterial inoculants hold promise for enhancing the sustainability and profitability of tomato cultivation in macro-tunnel systems. This study aimed to evaluate the technical and economic viability of applying Gluconacetobacter diazotrophicus to tomato production. The separate addition of native G. diazotrophicus GIBI025 and GIBI029 isolates and a commercial inoculant containing Azotobacter chrococcum and Azospirillium sp. was evaluated at a rate of 1 × 10⁸ CFU·mL⁻¹ without nitrogen addition. Conventional fertilization treatment with no bacteria added and 100%-nitrogen fertilization relative to crop requirements (added as MAP and urea) was also assessed. The treatments were evaluated within the macro-tunnel production system. The experiment utilized a completely randomized block design with four replications per treatment, and each experimental unit consisted of 20 plants. The yield (kg·ha⁻¹) was calculated and economic assessment was performed. The results show that native G. diazotrophicus isolates in tomato cultivation under the macro-tunnel production system improved its economic viability, achieving yields up to 95,501 kg·ha⁻¹ without the addition of nitrogenous fertilizers. This research reveals benefit–cost ratios achieving 1.57 and net incomes reaching 16,707 US dollars per hectare. This work demonstrated that the native isolates assessed may be used in the pursuit of more integrated, sustainable, and competitive cultural practices. Full article

Gluconacetobacter diazotrophicus is a diazotrophic endophytic bacterium that promotes the growth and development of several plant species. However, the molecular mechanisms activated during plant response to this bacterium remain unclear. Here, we used the RNA-seq approach to understand better the effect of G. diazotrophicus PAL5 on the transcriptome of shoot and root tissues of Arabidopsis thaliana. G. diazotrophicus colonized A. thaliana roots and promoted growth, increasing leaf area and biomass. The transcriptomic analysis revealed several differentially expressed genes (DEGs) between inoculated and non-inoculated plants in the shoot and root tissues. A higher number of DEGs were up-regulated in roots compared to shoots. Genes up-regulated in both shoot and root tissues were associated with nitrogen metabolism, production of glucosinolates and flavonoids, receptor kinases, and transcription factors. In contrast, the main groups of down-regulated genes were associated with pathogenesis-related proteins and heat-shock proteins in both shoot and root tissues. Genes encoding enzymes involved in cell wall biogenesis and modification were down-regulated in shoots and up-regulated in roots. In contrast, genes associated with ROS detoxification were up-regulated in shoots and down-regulated in roots. These results highlight the fine-tuning of the transcriptional regulation of A. thaliana in response to colonization by G. diazotrophicus PAL5. Full article

As agricultural practices become more sustainable, adopting more sustainable practices will become even more relevant. Searching for alternatives to chemical compounds has been the focus of numerous studies, and bacteriocins are tools with intrinsic biotechnological potential for controlling plant diseases. We continued to explore the biotechnological activity of the bacteriocin Gluconacin from Gluconacetobacter diazotrophicus, PAL5 strain, by investigating this protein’s antagonism against important tomato phytopathogens and demonstrating its effectiveness in reducing bacterial spots caused by Xanthomonas perforans. In addition to this pathogen, the bacteriocin Gluconacin demonstrated bactericidal activity in vitro against Ralstonia solanacearum and Pseudomonas syringae pv. tomato, agents that cause bacterial wilt and bacterial spots, respectively. Bacterial spot control tests showed that Gluconacin reduced disease severity by more than 66%, highlighting the biotechnological value of this peptide in ecologically correct formulations. Full article

Gluconacetobacter diazotrophicus is an endophytic bacterium that has shown important plant growth-promoting properties. During the growth of G. diazotrophicus with high carbon source concentrations, organic acids are produced, and pH decreases, thus inhibiting biomass growth. The objective of this work was to design a sucrose-based medium and perform a kinetic analysis of the batch submerged cultivation of this bacterium in a 3 L stirred-tank bioreactor without pH control. A mathematical model was proposed for representing G. diazotrophicus concentration, considering the inhibitory effect of hydrogen ion concentration. It comprises a biomass growth model, a specific growth rate expression that accounts for the inhibitory effect of hydrogen concentration, and a hydrogen model that represents the relationship between hydrogen and biomass concentrations. The sucrose-based medium proved its suitability for G. diazotrophicus growth. A higher biomass concentration (1.10 g/L) was obtained in a modified LGI-P medium containing 30 g/L sucrose with a three-fold increase in biomass production relative to the initial inoculation. The model allowed a satisfactory description of the experimental data obtained, and it could be used to design a cultivation strategy to maximize biomass production leading to the production of an alternative microbial inoculant for plant growth promotion of economically important crops. Full article

Gluconacetobacter diazotrophicus has been the focus of several studies aiming to understand the mechanisms behind this endophytic diazotrophic bacterium. The present study is the first global analysis of the early transcriptional response of exponentially growing G. diazotrophicus to iron, an essential cofactor for many enzymes involved in various metabolic pathways. RNA-seq, targeted gene mutagenesis and computational motif discovery tools were used to define the G. diazotrophicusfur regulon. The data analysis showed that genes encoding functions related to iron homeostasis were significantly upregulated in response to iron limitations. Certain genes involved in secondary metabolism were overexpressed under iron-limited conditions. In contrast, it was observed that the expression of genes involved in Fe-S cluster biosynthesis, flagellar biosynthesis and type IV secretion systems were downregulated in an iron-depleted culture medium. Our results support a model that controls transcription in G. diazotrophicus by fur function. The G. diazotrophicusfur protein was able to complement an E. colifur mutant. These results provide new insights into the effects of iron on the metabolism of G. diazotrophicus, as well as demonstrate the essentiality of this micronutrient for the main characteristics of plant growth promotion by G. diazotrophicus. Full article

The growth-promoting effects of Gluconacetobacter diazotrophicus inoculation on the leaf lettuce (Lactuca sativa L.) cultivars “Black Seeded Simpson” and “Bibb/Limestone” were investigated. Plants of each cultivar were grown hydroponically in Kratky jars in a growth chamber-controlled environment in a completely randomized factorial design with three or four replications. Each experiment was repeated once. Factors were (1) with or without inoculant and (2) seven levels of nitrogen (N) fertilization ranging from deficient (37.5 mg L⁻¹ N) to excessive (172.5 mg L⁻¹ N). The shoot, root, and total biomass accumulation, nitrogen density, and carbon/nitrogen (C/N) ratios were measured for each variety. Black Seeded Simpson demonstrated a shifting of production towards aerial tissues, with significantly greater shoot production and reduced root production. The observed increase in shoot biomass was greatest at the slightly deficient N rate of 105 mg L⁻¹ N where inoculated plants produced 14.8% more than uninoculated plants. Lower N density and higher C/N ratios in inoculated shoot tissues indicate greater N use efficiency. Bibb/Limestone responded to inoculation with an average increase of 10.9% in shoot production and with greater root biomass. Bibb/Limestone also exhibited lower N density in inoculated shoot tissues with a corresponding increase in the C/N ratio. For growers looking to maximize lettuce yields, G. diaz inoculation may present a beneficial additive to the growing system by increasing leaf yields while not increasing N fertilizer requirements. Full article

The biosynthesis of exopolysaccharides (EPSs) is essential for endophytic bacterial colonisation in plants bacause this exopolymer both protects bacterial cells against the defence and oxidative systems of plants and acts on the plant colonisation mechanism in Gluconacetobacter diazotrophicus. The pathway involved in the biosynthesis of bacterial EPS has not been fully elucidated, and several areas related to its molecular regulation mechanisms are still lacking. G. diazotrophicus relies heavily on EPS for survival indirectly by protecting plants from pathogen attack as well as for endophytic maintenance and adhesion in plant tissues. Here, we report that EPS from G. diazotrophicus strain Pal5 is a signal polymer that controls its own biosynthesis. EPS production depends on a bacterial tyrosine (BY) kinase (Wzc) that consists of a component that is able to phosphorylate a glycosyltranferase or to self-phosphorylate. EPS interacts with the extracellular domain of Wzc, which regulates kinase activity. In G. diazotrophicus strains that are deficient in EPS production, the Wzc is rendered inoperative by self-phosphorylation. The presence of EPS promotes the phosphorylation of a glycosyltransferase in the pathway, thus producing EPS. Wzc-mediated self-regulation is an attribute for the control of exopolysaccharide biosynthesis in G. diazotrophicus. Full article

Industrial hemp (Cannabis sativa L.) is a multipurpose plant used in several fields. Several phytopathogens attack hemp crops. Fusarium oxysporum is a common fungal pathogen that causes wilt disease in nurseries and in field cultivation and causes high losses. In the present study, a pathogenic strain belonging to F. oxysporum f. sp. cannabis was isolated from a plant showing Fusarium wilt. After isolation, identification was conducted based on morphological and molecular characterizations and pathogenicity tests. Selected plant growth-promoting bacteria with interesting biocontrol properties—Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and Burkholderia ambifaria—were tested against this pathogen. In vitro antagonistic activity was determined by the dual culture method. Effective strains (in vitro inhibition > of 50%) G. diazotrophicus, H. seropedicae and B. ambifaria were combined in a consortium and screened for in planta antagonistic activity in pre-emergence (before germination) and post-emergence (after germination). The consortium counteracted Fusarium infection both in pre-emergence and post-emergence. Our preliminary results show that the selected consortium could be further investigated as an effective biocontrol agent for the management of this pathogen. Full article

Plant growth promoting endophytic bacteria, which can fix nitrogen, plays a vital role in plant growth promotion. Previous authors have evaluated the effect of Gluconacetobacter diazotrophicus Pal5 inoculation on plants subjected to different sources of abiotic stress on an individual basis. The present study aimed to appraise the effect of G. diazotrophicus inoculation on the amelioration of the individual and combined effects of drought and nitrogen stress in maize plants (Zea mays L.). A pot experiment was conducted whereby treatments consisted of maize plants cultivated under drought stress, in soil with a low nitrogen concentration and these two stress sources combined, with and without G. diazotrophicus seed inoculation. The inoculated plants showed increased plant biomass, chlorophyll content, plant nitrogen uptake, and water use efficiency. A general increase in copy numbers of G. diazotrophicus, based on 16S rRNA gene quantification, was detected under combined moderate stress, in addition to an increase in the abundance of genes involved in N fixation (nifH). Endophytic colonization of bacteria was negatively affected by severe stress treatments. Overall, G. diazotrophicus Pal5 can be considered as an effective tool to increase maize crop production under drought conditions with low application of nitrogen fertilizer. Full article

Background: Inoculation with Gluconacetobacter diazotrophicus has shown to influence root development in red rice plants, and more recently, the induced systemic tolerance (IST) response to drought was also demonstrated. The goal of this study was to evaluate the inoculation effect of G. diazotrophicus strain Pal5 on the amelioration of drought stress and root development in red rice (Oryza sativa L.). Methods: The experimental treatments consist of red rice plants inoculated with and without strain Pal5 in presence and absence of water restriction. Physiological, biochemical, and molecular analyses of plant roots were carried out, along with measurements of growth and biochemical components. Results: The plants showed a positive response to the bacterial inoculation, with root growth promotion and induction of tolerance to drought. An increase in the root area and higher levels of osmoprotectant solutes were observed in roots. Bacterial inoculation increased the drought tolerance and positively regulated certain root development genes against the water deficit in plants. Conclusion: G. diazotrophicus Pal5 strain inoculation favored red rice plants by promoting various root growth and developmental mechanisms against drought stress, enabling root development and improving biochemical composition. Full article

Gluconacetobacter diazotrophicus PAL5 (GDI) is an endophytic bacterium with potential biotechnological applications in industry and agronomy. The recent description of its complete genome and its principal metabolic enzymes suggests that glucose metabolism is accomplished through the pentose phosphate pathway (PPP); however, the enzymes participating in this pathway have not yet been characterized in detail. The objective of the present work was to clone, purify, and biochemically and physicochemically characterize glucose-6-phosphate dehydrogenase (G6PD) from GDI. The gene was cloned and expressed as a tagged protein in E. coli to be purified by affinity chromatography. The native state of the G6PD protein in the solution was found to be a tetramer with optimal activity at pH 8.8 and a temperature between 37 and 50 °C. The apparent Km values for G6P and nicotinamide adenine dinucleotide phosphate (NADP⁺) were 63 and 7.2 μM, respectively. Finally, from the amino acid sequence a three-dimensional (3D) model was obtained, which allowed the arrangement of the amino acids involved in the catalytic activity, which are conserved (RIDHYLGKE, GxGGDLT, and EKPxG) with those of other species, to be identified. This characterization of the enzyme could help to identify new environmental conditions for the knowledge of the plant–microorganism interactions and a better use of GDI in new technological applications. Full article