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An inoculant containing selected bacterial strains can be easily applied during the nursery please process, but in this case, substrate composition can affect its survival and, in consequence, bacterial colonization. The aim of this study was to evaluate the survival of five diazotrophic bacterial strains/species applied individually on 13 different substrates used in sugarcane seedling nurseries considering an active population higher than 105 cells g-1. In addition, one experiment was performed using two commercial substrates, coconut fiber, and Multiplant™, inoculated or not with a mixture of five bacterial strains to evaluate plant growth after 30 days. These strains are combined inoculants selected for sugarcane acting as plant growth promoters. Bacterial counts were determined every seven days using the Most Probable Number technique with four different semi-solid N-free media specific for each strain tested over 35 days. The survival order, independent of the substrate tested, was: Paraburkholderia tropica Pt-PPe8T > Nitrospirillum amazonense Na-CBAMc > Herbaspirillum seropedicae Hs-HRC54 = H. rubrisubalbicans Hr-HCC103 > Gluconacetobacter diazotrophicus Gd-PAL5T. All tested substrates influenced the bacterial survival, especially after 21 days of incubation. The population size can be partially controlled by the substrate pH and stimulated by the addition of slow-release fertilizer. Besides the differences in the bacterial population present in the two commercial substrates, plant growth was found to be stimulated by the inoculated bacteria, depending on the substrate and its sugarcane cultivar tested. The selection of a substrate used to produce new plantlets of sugarcane can contribute to bacterial survival and improve bacterial colonization.

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Agave lechuguilla is a widely distributed plant in arid ecosystems. It has been suggested that its microbiome is partially responsible for its great adaptability to the oligotrophic environments of the Chihuahuan Desert. To lead the recruitment of beneficial rhizobacteria, the root exudates are essential; however, the amino acids contained within these compounds had been largely overlooked. Thus, we investigated how the variations of amino acids in the rhizosphere at different growth stages of A. lechuguilla affect the rhizobacterial community composition, its functions, and activity of the beneficial bacteria. In this regard, it was found that arginine and tyrosine were related to the composition of the rhizobacterial community associated to A. lechuguilla, where the most abundant genera were from the phylum Proteobacteria and Bacteroidetes. Moreover, Firmicutes was largely represented by Bacillus in the phosphorus-mineralizing bacteria community, which may indicate its great distribution and versatility in the harsh environments of the Chihuahuan Desert. In contrast, we found a high proportion of Unknown taxa of nitrogen-fixing bacteria, reflecting the enormous diversity in the rhizosphere of these types of plants that remains to be explored. This work also reports the influence of micronutrients and the amino acids methionine and arginine over the increased activity of the nitrogen-fixing and phosphorus-mineralizing bacteria in the rhizosphere of lechuguillas. In addition, the results highlight the multiple beneficial functions present in the microbiome that could help the host to tolerate arid conditions and improve nutrient availability.

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Sugarcane is an economically important crop that is used for the production of fuel ethanol. Diazotrophic bacteria have been isolated from sugarcane tissues, without causing visible plant anatomical changes or disease symptoms. These bacteria can be beneficial to the plant by promoting root growth and an increase in plant yield. Different rates of Biological Nitrogen Fixation (BNF) were observed in different genotypes. The aim of this work was to conduct a comprehensive molecular and physiological analysis of two model genotypes for contrasting BNF efficiency in order to unravel plant genes that are differentially regulated during a natural association with diazotrophic bacteria. A next-generation sequencing of RNA samples from the genotypes SP70-1143 (high-BNF) and Chunee (low-BNF) was performed. A differential transcriptome analysis showed that several pathways were differentially regulated among the two BNF-contrasting genotypes, including nitrogen metabolism, hormone regulation and bacteria recognition. Physiological analyses, such as nitrogenase and GS activity quantification, bacterial colonization, auxin response and root architecture evaluation, supported the transcriptome expression analyses. The differences observed between the genotypes may explain, at least in part, the differences in BNF contributions. Some of the identified genes might be involved in key regulatory processes for a beneficial association and could be further used as tools for obtaining more efficient BNF genotypes.

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This research was aimed at obtaining varieties of soybean adaptive to acid soils and to obtain Plant Growth-Promoting Rhizobacteria (PGPR) isolates that can improve the agronomic characteristics of soybean and increase the ultisols fertility. This research was conducted in two-stages research on Sampali Village, Percut Sei Tuan sub-District, Deli Serdang District, North Sumatra Province, Indonesia from August 2019 until March 2020. The first stage (adaptive varieties screening) using the non-factorial Randomized Block Design (RBD) with the varieties of Argomulyo, Wilis, Kaba, Dena-1, Devon-1, Dega-1, Demas-1, Burangrang, Detam-1, and Kipas Merah. The second stage (application of PGPR isolates singly and in combinations) using the factorial RBD, the first factor of applicative single and the combination of PGPR isolates, the second factor of adaptive varieties including Detam-1 and Wilis. Data were analyzed with ANOVA and followed by DMRT at P<0.05. The results showed that the Detam-1 and Wilis varieties had significantly higher yield per plant of 14.73 g and 14.54 g, respectively, than other varieties. The applications of a single and combination of PGPR isolates significantly increased the number of branches, stem diameter, plant height, yield per plant, soil pH, organic-C, available-P, and total-N and decreased the soil C/N. The Detam-1 variety had the higher in yield per plant compared to Wilis variety. The isolates combination of Rhizobium leguminosarum+Rhizobium sp2+Bacillus sp+Burkholderia sp for Detam-1 and Wilis varieties can be recommended to support the growth and yield of soybean on ultisols.

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Abstract: Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum seropedicae occurred only in stems. Except to IPA-CF45A, all isolates were nitrogenase positive. All endophytes exhibit production of indol 3-acetic acid. Over 50% of endophytes solubilize phosphate, release N-acyl homoserine lactones, and present the activity of 1-aminocyclopropane-1-carboxylic acid deaminase, catalase, lipase and protease. The network analysis showed that isolates belonged to Burkholderia, Herbaspirillum, and Methylobacterium interact with Bacillus. Bacterial endophytes exhibited distinct morphological, physiological, and PGP traits that are useful for sustainable agriculture, highlighting the isolates IPA-CC33, IPA-CF65, IPA-CC9 and IPA-CF27. Further studies on the effects of these diazotrophic endophytes and their potential for providing microbial inoculants for improving sugarcane fields will provide valuable information to maintain the sustainability and environment quality.

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Agave lechuguilla has one of the widest distributions among other agave species in the Chihuahuan Desert. Their capacity to grow in poorly developed soils and harsh conditions has been related to their association with plant growth-promoting rhizobacteria. In this work, we explored how soil properties and plant growth stage influence the composition of the rhizobacterial communities, their interactions, and the enzymatic activity and abundance of nitrogen-fixing bacteria and organic phosphorus-mineralizing bacteria in two subregions of the Chihuahuan Desert. We found that mature plants of lechuguilla stimulated the activity and abundance of nutrient-improvement rhizobacteria, and these soil samples had a higher content of total organic carbon, ammonium (NH4) and nitrite + nitrate (NO2+NO3). Nutrient availability seems to be an essential driver of the bacterial community's structure since the genera with more connections (hubs) were those with known mechanisms related to the availability of nutrients, such as env. OPS17 (Bacteroidetes), Gemmatimonadaceae uncultured, S0134terrestrial group, BD211terrestrial group (Gemmatimonadetes), Chthoniobacteracea and Candidatus Udaeobacter (Verrucomicrobia). This work shows that the late growth stages of lechuguilla recruit beneficial bacteria that favor its establishment and tolerance to harsh conditions of the arid lands.

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The increasing use of chemical fertilizers causes the loss of natural biological nitrogen fixation in soils, water eutrophication and emits more than 300 Mton CO2 per year. It also limits the success of external bacterial inoculation in the soil. Nitrogen fixing bacteria can be inhibited by the presence of ammonia as its presence can inhibit biological nitrogen fixation. Two aerobic sludges from wastewater treatment plants (WWTP) were exposed to high ammonium salts concentrations (>450 mg L-1 and >2 dS m-1). Microbial analysis after treatment through 16S pyrosequencing showed the presence of Fluviicola sp. (17.70%), a genus of the Clostridiaceae family (11.17%), and Azospirillum sp. (10.42%), which were present at the beginning with lower abundance. Denaturing gradient gel electrophoresis (DGGE) analysis based on nifH genes did not show changes in the nitrogen-fixing population. Nitrogen-Fixing Bacteria (NFB) were identified and associated with other microorganisms involved in the nitrogen cycle, presumably for survival at extreme conditions. The potential use of aerobic sludges enriched with NFB is proposed as an alternative to chemical fertilizer as this bacteria could supplement nitrogen to the plant showing competitive results with chemical fertilization.

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Basic knowledge of transcriptional regulation is needed to understand the mechanisms governing biological processes, i.e., nitrogen fixation by Rhizobiales bacteria in symbiosis with leguminous plants. The RhizoBindingSites database is a computer-assisted framework providing motif-gene-associated conserved sequences potentially implicated in transcriptional regulation in nine symbiotic species. A dyad analysis algorithm was used to deduce motifs in the upstream regulatory region of orthologous genes, and only motifs also located in the gene seed promoter with a p-value of 1e-4 were accepted. A genomic scan analysis of the upstoream sequences with these motifs was performed. These predicted binding sites were categorized according to low, medium and high homology between the matrix and the upstream regulatory sequence. On average, 62.7% of the genes had a motif, accounting for 80.44% of the genes per genome, with 19613 matrices (a matrix is a representation of a motif). The RhizoBindingSites database provides motif and gene information, motif conservation in the order Rhizobiales, matrices, motif logos, regulatory networks constructed from theoretical or experimental data, a criterion for selecting motifs and a guide for users. The RhizoBindingSites database is freely available online at rhizobindingsites.ccg.unam.mx.

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Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing Azospirillum brasilense. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications.

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RESUMEN Gluconacetobacter diazotrophicus es una bacteria endófita promotora del crecimiento vegetal utilizada como inoculante microbiano en diferentes cultivos agrícolas. El objetivo del presente trabajo fue aplicar diferentes modelos matemáticos para representar su crecimiento en un cultivo sumergido por lotes empleando un biorreactor de 3 L y usando melazas de caña y sacarosa como fuente de energía. Se obtuvo el perfil temporal de pH, biomasa celular y azúcares totales. Se compararon los modelos estudiados por calidad de ajuste y complejidad y se realizó un análisis de sensibilidad paramétrica. Se consideraron modelos de cuatro y cinco parámetros con expresiones que incluyen efectos de inhibición por sustrato y por biomasa. El modelo con mayor calidad de ajuste fue el de Herbert-Pirt-Contois con coeficientes de determinación para biomasa y sustrato de 0,888 y 0,425 respectivamente. Estos valores indican una mayor correspondencia de los datos experimentales de biomasa con los datos calculados por el modelo, en comparación con los resultados obtenidos para azúcares totales para los que esta correspondencia fue menor. Este modelo generó la mejor combinación de calidad de ajuste y complejidad según el criterio de información de Akaike. El estudio cinético desarrollado permitió observar un comportamiento bifásico en la etapa de crecimiento de la bacteria cuando se cultiva en melaza y un efecto de limitación de su crecimiento por la biomasa. Los resultados obtenidos proporcionan una descripción matemática útil para el diseño, escalamiento y operación de un futuro proceso de producción de un inoculante microbiano a base de la bacteria G. diazotrophicus.

ABSTRACT Gluconacetobacter diazotrophicus is a plant-growth promoting endophytic bacterium used as a microbial inoculant for different crops. The objective of this work was to apply different mathematical models to represent its growth in a batch submerged culture employing a 3-L bioreactor and using sugarcane molasses and sucrose as energy sources. The time profile of pH, cell biomass, and total sugars was obtained. Models studied were compared considering their fit quality and complexity, and a parametric sensitivity analysis was performed. Four- and five-parameter models with expressions involving substrate and biomass inhibition effects were considered. The Herbert-Pirt-Contois model achieved the highest fit quality with determination coefficients of 0.888 and 0.425 for biomass and substrate, respectively. These values indicate a higher correspondence between the experimental data of biomass concentration and the data calculated by the model, compared to results obtained for total sugars for which this correspondence was lower. This model reached the best combination considering the fit quality and complexity according to the Akaike's information criterion. The kinetic study performed enabled to observe a bi-phasic behavior in the growth stage of the bacterium when grown on molasses, and a growth limitation effect due to biomass concentration. The outcomes obtained provide a mathematical description useful for design, scale-up, and operation of a future process for the production of a microbial inoculant based on G. diazotrophicus.

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The response of sugarcane (Saccharum spp.) families to the inoculation with plant growth-promoting bacteria (PGPB) may show positive or negative interaction values depending on the plant genotype, allowing the selection of responsive genotypes to the association. Based on these precepts, this study aimed to evaluate the response of 27 sugarcane families treated with two PGPB-based inoculants in two sugarcane cycles: plant-cane and first ratoon. Four months after germination, 27 families from crosses between clones of sugarcane were: (T0) not inoculated; (T1) inoculated with Triazo, which consists of a mixture of the strains Abv5, Abv6, and Abv7 of Azospirillum brasilense; and (T2) inoculated with an inoculant called Mix, which contains five strains of PGPB (Gluconacetobacter diazotrophicus Pal5, Azospirillum amazonense CBAmC, Burkholderia tropica Ppe8, Herbaspirillum rubrisubalbicans HCc103, and Herbaspirillum seropedicae HRC54). The productivity traits evaluated were the number of stalks, average height, stalk diameter, and Brix. The correlation values between the two cycles were 0.87 for the number of stalks, 0.70 for average height, 0.48 for stalk diameter, and 0.63 for Brix. Differences were observed between treatments for average height, stalk diameter, and Brix. Differences were also observed between families for all the studied traits. Brix was the only parameter that...(AU)

A resposta de famílias de cana-de-açúcar (Saccharum spp.) à inoculação com bactérias promotoras de crescimento vegetal (PGPB) pode apresentar valores positivos e negativos de interação, o que possibilita a seleção genótipos responsivos à associação. O objetivo do presente estudo foi avaliar a resposta de 27 famílias de cana-de-açúcar tratadas com dois diferentes inoculantes a base de PGPB em dois ciclos de cultivo, cana-planta e cana-soca. Famílias provenientes de cruzamentos biparentais receberam três diferentes tratamentos: (T0) não inoculadas; (T1) inoculadas com Triazo (composto pelas estirpes Abv5, Abv6 e Abv7 de Azospirillum brasilense); (T2) inoculadas com um Mix contendo cinco diferentes espécies de PGPB (Gluconacetobacter diazotrophicus Pal5, Azospirillum amazonense CBAmC, Burkholderia tropica Ppe8, Herbaspirillum rubrisubalbicans HCc103 e Herbaspirillum seropedicae HRC54). Foram avaliadas as seguintes variáveis por planta: número de colmos, altura média, diâmetro médio e Brix. Os resultados mostraram que o valor de correlação entre os dois ciclos foi de 0,87, 0,70, 0,48 e 0,63, para número de colmos, altura, diâmetro e Brix, respectivamente. Houve efeito significativo do tratamento para as variáveis, altura e diâmetro médio da planta e Brix, assim como diferença entre as famílias para todas as variáveis estudadas. A variável Brix foi a única que apresentou interação...(AU)

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The response of sugarcane (Saccharum spp.) families to the inoculation with plant growth-promoting bacteria (PGPB) may show positive or negative interaction values depending on the plant genotype, allowing the selection of responsive genotypes to the association. Based on these precepts, this study aimed to evaluate the response of 27 sugarcane families treated with two PGPB-based inoculants in two sugarcane cycles: plant-cane and first ratoon. Four months after germination, 27 families from crosses between clones of sugarcane were: (T0) not inoculated; (T1) inoculated with Triazo, which consists of a mixture of the strains Abv5, Abv6, and Abv7 of Azospirillum brasilense; and (T2) inoculated with an inoculant called Mix, which contains five strains of PGPB (Gluconacetobacter diazotrophicus Pal5, Azospirillum amazonense CBAmC, Burkholderia tropica Ppe8, Herbaspirillum rubrisubalbicans HCc103, and Herbaspirillum seropedicae HRC54). The productivity traits evaluated were the number of stalks, average height, stalk diameter, and Brix. The correlation values between the two cycles were 0.87 for the number of stalks, 0.70 for average height, 0.48 for stalk diameter, and 0.63 for Brix. Differences were observed between treatments for average height, stalk diameter, and Brix. Differences were also observed between families for all the studied traits. Brix was the only parameter that...

A resposta de famílias de cana-de-açúcar (Saccharum spp.) à inoculação com bactérias promotoras de crescimento vegetal (PGPB) pode apresentar valores positivos e negativos de interação, o que possibilita a seleção genótipos responsivos à associação. O objetivo do presente estudo foi avaliar a resposta de 27 famílias de cana-de-açúcar tratadas com dois diferentes inoculantes a base de PGPB em dois ciclos de cultivo, cana-planta e cana-soca. Famílias provenientes de cruzamentos biparentais receberam três diferentes tratamentos: (T0) não inoculadas; (T1) inoculadas com Triazo (composto pelas estirpes Abv5, Abv6 e Abv7 de Azospirillum brasilense); (T2) inoculadas com um Mix contendo cinco diferentes espécies de PGPB (Gluconacetobacter diazotrophicus Pal5, Azospirillum amazonense CBAmC, Burkholderia tropica Ppe8, Herbaspirillum rubrisubalbicans HCc103 e Herbaspirillum seropedicae HRC54). Foram avaliadas as seguintes variáveis por planta: número de colmos, altura média, diâmetro médio e Brix. Os resultados mostraram que o valor de correlação entre os dois ciclos foi de 0,87, 0,70, 0,48 e 0,63, para número de colmos, altura, diâmetro e Brix, respectivamente. Houve efeito significativo do tratamento para as variáveis, altura e diâmetro médio da planta e Brix, assim como diferença entre as famílias para todas as variáveis estudadas. A variável Brix foi a única que apresentou interação...

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Plant growth-promoting bacteria (PGPB) and humic acids (HA) have been used as biostimulants in field conditions. The complete genomic and proteomic transcription of Herbaspirillum seropedicae and Gluconacetobacter diazotrophicus is available but interpreting and utilizing this information in the field to increase crop performance is challenging. The identification and characterization of metabolites that are induced by genomic changes may be used to improve plant responses to inoculation. The objective of this study was to describe changes in sugarcane metabolic profile that occur when HA and PGPB are used as biostimulants. Inoculum was applied to soil containing 45-day old sugarcane stalks. One week after inoculation, the methanolic extracts from leaves were obtained and analyzed by gas chromatography coupled to time-of-flight mass spectrometry; a total of 1,880 compounds were observed and 280 were identified in all samples. The application of HA significantly decreased the concentration of 15 metabolites, which generally included amino acids. HA increased the levels of 40 compounds, and these included metabolites linked to the stress response (shikimic, caffeic, hydroxycinnamic acids, putrescine, behenic acid, quinoline xylulose, galactose, lactose proline, oxyproline and valeric acid) and cellular growth (adenine and adenosine derivatives, ribose, ribonic acid and citric acid). Similarly, PGPB enhanced the level of metabolites identified in HA-treated soils; e.g., 48 metabolites were elevated and included amino acids, nucleic acids, organic acids, and lipids. Co-inoculation (HA+PGPB) boosted the level of 110 metabolites with respect to non-inoculated controls; these included amino acids, lipids and nitrogenous compounds. Changes in the metabolic profile induced by HA+PGPB influenced both glucose and pentose pathways and resulted in the accumulation of heptuloses and riboses, which are substrates in the nucleoside biosynthesis and shikimic acid pathways. The mevalonate pathway was also activated, thus increasing phytosterol synthesis. The improvement in cellular metabolism observed with PGPB+HA was compatible with high levels of vitamins. Glucuronate and amino sugars were stimulated in addition to the products and intermediary compounds of tricarboxylic acid metabolism. Lipids and amino acids were the main compounds induced by co-inoculation in addition to antioxidants, stress-related metabolites, and compounds involved in cellular redox. The primary compounds observed in each treatment were identified, and the effect of co-inoculation (HA+PGPB) on metabolite levels was discussed.

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Eucalyptus plantations offer a cost-effective and renewable source of raw material. There is substantial interest in improving forestry production, especially through sustainable strategies such as the use of plant growth-promoting bacteria. However, little is known about Eucalyptus microbiology. In this study, the endophytic bacterial community was assessed in Eucalyptus urograndis roots using culture-dependent and culture-independent techniques with plants grown under different conditions. Three phyla accounted for approximately 95% of the community, with Actinobacteria corresponding to approximately 59%. This contrasts with previous studies in which Actinobacteria accounted for only 5 to 10%. Our data also revealed a high diversity of bacteria, with 359 different genera but a high level of dominance. Six genera, Mycobacterium, Bradyrhizobium, Streptomyces, Bacillus, Actinospica, and Burkholderia, accounted for more than 50% of the classified sequences. We observed a significant influence of the treatments on some genera, causing changes in the bacterial community structure. The obtained data also suggest that Eucalyptus may benefit from biological nitrogen fixation, with many abundant genera being closely related to nitrogen-fixing bacteria. Using N-depleted media, we also cultured 95 bacterial isolates, of which 24 tested positive for the nifH gene and were able to maintain growth without any N source in the medium.

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The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that coinoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution.

El objetivo de esta investigación fue evaluar si la aplicación de 2 (rizo)bacterias promotoras del crecimiento vegetal podría reducir la dosis de fertilizante nitrogenado en el cultivo de algodón. Se usaron las cepas Azotobacter chroococcum AC1 y AC10 por su habilidad para promover la germinación de semillas y el crecimiento del algodonero. Estos microorganismos fueron caracterizados sobre la base de sus actividades de promoción del crecimiento vegetal. Luego se realizó un estudio de invernadero con plantas de algodón para evaluar la capacidad de promoción del crecimiento vegetal de dichas cepas con dosis reducidas de urea. Los resultados revelaron que ambas cepas son capaces de fijar nitrógeno, solubilizar fósforo, sintetizar compuestos indólicos y producir enzimas hidrolíticas. Después de 12 semanas, el experimento de invernadero permitió observar que el crecimiento del algodón fue influido positivamente por la inoculación bacteriana con respecto a la fertilización química. En particular, se evidenció que la inoculación microbiana impactó más en la biomasa vegetal (p<0,05) que en el contenido de nitrógeno. Curiosamente, la coinoculación exhibió un mayor efecto positivo sobre los parámetros de crecimiento en comparación con la inoculación simple. Además, se observaron resultados similares, sin diferencias estadísticamente significativas, entre la coinoculación bacteriana más del 50% de urea y el 100% de fertilización. Estos hallazgos indican que la coinoculación de las cepas de A. chroococcum AC1 y AC10 permitiría reducir las dosis de fertilización nitrogenada del cultivo de arroz en hasta el 50% y aliviar, de esta manera, el deterioro ambiental relacionado con la contaminación por N.

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Although Azospirillum strains used in commercial inoculant formulations presents diazotrophic activity, it has been reported that their ability to produce phytohormones plays a pivotal role in plant growth-promotion, leading to a general recommendation of its use in association with regular N-fertilizer doses. In addition, a high variability in the effectiveness of Azospirillum inoculants is still reported under field conditions, contributing to the adoption of the inoculation technology as an additional management practice rather than its use as an alternative practice to the use of chemical inputs in agriculture. To investigate whether the content of stress-resistance biopolymers would improve the viability and performance of Azospirillum inoculants when used as substitute of N-fertilizers, biomass of A. brasilense strain Ab-V5 enriched in exopolysaccharides (EPS) and polyhydroxybutirate (PHB) was produced using a new culture medium developed by factorial mixture design, and the effectiveness of resulting inoculants was evaluated under field conditions. The culture medium formulation extended the log phase of A. brasilense cultures, which presented higher cell counts and increased EPS and PHB contents than observed in the cultures grown in the OAB medium used as control. An inoculation trial with maize conducted under greenhouse conditions and using the biopolymers-enriched Ab-V5 cells demonstrated the importance of EPS and PHB to the long term bacterial viability in soil and to the effectiveness of inoculation. The effectiveness of liquid and peat inoculants prepared with Ab-V5 cells enriched with EPS and PHB was also evaluated under field conditions, using maize as target crop along different seasons, with the inoculants applied directly over seeds or at topdressing under limiting levels of N-fertilization. No additive effect on yield resulted from inoculation under high N fertilizer input, while inoculated plants grown under 80% reduction in N fertilizer showed yields at levels compared to fully fertilized plants, regardless the inoculation method. The presented data highlights the feasibility to partially substitute the N-fertilizer demand in non-legume crops using high-quality inoculant formulations, prepared with diazotrophic bacteria enriched with stress-resistance biopolymers that confer increased viability an effectiveness to the bacterial cells.

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The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that co-inoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution.

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RESUMO

Microbial solubilisation of low soluble inorganic phosphates is an important process contributing for the phosphorus available to plants in tropical soils. This study evaluates the ability of inoculant strains for tropical legumes to solubilise inorganic phosphates of low solubility that are found in tropical soils. Seven strains of Leguminosae nodulating bacteria (LNB) were compared with one another and with a non-nodulating positive control, Burkholderia cepacia (LMG 1222T). Four of the strains are used as inoculants for cowpeas (Vigna unguiculata) (Bradyrhizobium sp. UFLA 03-84; Bradyrhizobium elkani INPA 03-11B and Bradyrhizobium japonicum BR3267) or for common beans (Phaseolus vulgaris) (Rhizobium tropici CIAT 899T). Rhizobium etli UFLA 02-100 and Rhizobium leguminosarum 316C10a are also efficient nodulators of beans and Cupriavidus taiwanensis LMG 19424T nodulates on Mimosa pudica. Two experiments, with solid and liquid media, were performed to determine whether the strains were able to solubilise CaHPO4, Al(H2PO4)3 or FePO4.2H2O. On solid GELP medium none of the strains dissolved FePO4.2H2O, but LMG 1222, UFLA 03-84 and CIAT 899 solubilised CaHPO4 particularly well. These strains, along with LMG 19424 and BR 3267, were also able to increase the solubility of Al(H2PO4)3. In liquid GELP medium, LMG 1222 solubilised all phosphate sources, but no legume nodulating strain could increase the solubility of Al(H2PO4)3. The strains CIAT 899 and UFLA 02-100 were the only legume nodulating bacteria able to solubilise the other phosphate sources in liquid media, dissolving both CaHPO4 and FePO4.2H2O. There was a negative correlation between the pH of the culture medium and the concentration of soluble phosphate when the phosphorus source was CaHPO4 or FePO4.2H2O. The contribution of these strains to increasing the phosphorus nutrition of legumes and non-legume plant species should be investigated further by in vivo experiments.

RESUMO

Microbial solubilisation of low soluble inorganic phosphates is an important process contributing for the phosphorus available to plants in tropical soils. This study evaluates the ability of inoculant strains for tropical legumes to solubilise inorganic phosphates of low solubility that are found in tropical soils. Seven strains of Leguminosae nodulating bacteria (LNB) were compared with one another and with a non-nodulating positive control, Burkholderia cepacia (LMG 1222T). Four of the strains are used as inoculants for cowpeas (Vigna unguiculata) (Bradyrhizobium sp. UFLA 03-84; Bradyrhizobium elkani INPA 03-11B and Bradyrhizobium japonicum BR3267) or for common beans (Phaseolus vulgaris) (Rhizobium tropici CIAT 899T). Rhizobium etli UFLA 02-100 and Rhizobium leguminosarum 316C10a are also efficient nodulators of beans and Cupriavidus taiwanensis LMG 19424T nodulates on Mimosa pudica. Two experiments, with solid and liquid media, were performed to determine whether the strains were able to solubilise CaHPO4, Al(H2PO4)3 or FePO4.2H2O. On solid GELP medium none of the strains dissolved FePO4.2H2O, but LMG 1222, UFLA 03-84 and CIAT 899 solubilised CaHPO4 particularly well. These strains, along with LMG 19424 and BR 3267, were also able to increase the solubility of Al(H2PO4)3. In liquid GELP medium, LMG 1222 solubilised all phosphate sources, but no legume nodulating strain could increase the solubility of Al(H2PO4)3. The strains CIAT 899 and UFLA 02-100 were the only legume nodulating bacteria able to solubilise the other phosphate sources in liquid media, dissolving both CaHPO4 and FePO4.2H2O. There was a negative correlation between the pH of the culture medium and the concentration of soluble phosphate when the phosphorus source was CaHPO4 or FePO4.2H2O. The contribution of these strains to increasing the phosphorus nutrition of legumes and non-legume plant species should be investigated further by in vivo experiments.

RESUMO

Distribution of potentially nitrogen-fixing bacteria and its relationship with physicochemical parameters in soils with three vegetation types in the southern Colombian Amazon region. Potentially nitrogen-fixing microaerobic and aerobic bacteria were isolated from several Colombian Amazon soils (forest, pastures and chagras) and two landscapes (floodable and non floodable areas). The abundance and distribution of bacteria were evaluated, as well as their relationship with soil physical and chemical characteristics. Landscape had a direct influence on the abundance of the microaerobic bacteria, with higher numbers in forest and pasture soils in non- floodable zones. The aerobic isolates (N=51) were grouped into 19 morphologies, with the highest numbers found in forest soil in floodable zones. A higher number of aerobic morphologies was shared among forest sites (Nonmetric Multidimensional Scaling and Analysis of Similarity p<0.05), and 40% of the distribution was explained by lime percentage and Al concentration. Rev. Biol. Trop. 57 (4): 915- 927. Epub 2009 December 01.

Se evaluó la abundancia y distribución de bacterias microaerófilas y aerobias potencialmente fijadoras de nitrógeno aisladas a partir de suelos bajo coberturas de bosque, pastizal y chagra en dos paisajes, terraza y llanura inundable, en el sur de trapecio amazónico. Se relacionó el recuento en placa en el medio Ashby de estas bacterias aerobias con las características físicas y químicas del suelo. Se encontró mayor abundancia de bacterias aerobias y microaerofilas en suelos bajo cobertura de pastizal. Se observó influencia directa del paisaje sobre la abundancia de bacterias microaerófilas registrando mayores recuentos los suelos bajo terraza en las coberturas de bosque y pastizal. Los aislamientos aerobios obtenidos (51) se agruparon en 19 morfologías, de las cuales se obtuvo mayor número en suelos bajo bosque y en el paisaje de llanura inundable. A través del análisis multidimensional no métrico (NMDS) y análisis de similaridades (ANOSIM) (p<0.05) se comprobó que entre los sitios de muestreo bajo la cobertura de bosque se comparten mayor número de morfologías de bacterias aerobias que bajo las demás coberturas. El programa BIOENV indicó que esta distribución fue explicada en un 40% por las variables % limo y Al (meq/100 g).

Assuntos