Inoculation effects of Nitrospirillum amazonense and biofertilizer in sugarcane / Efeitos da inoculação de Nitrospirillum amazonense e biofertilizante em cana-de-açúcar

In this study, we analyzed the hypothesis that the combination of Nitrospirillum amazonense strain BR11145 with biological fertilizer prepared using Microgeo® and native microbiome from location of the product application results in morphological and nutritional gains for the initial development of sugarcane plants and soil chemistry. For this purpose, pre-sprouted sugarcane seedlings were grown in a greenhouse mesocosm experiment using soil amended with nitrogen/phosphorus/potassium fertilizer. The experimental treatments consisted of: 1) biological fertilizer with the addition of N. amazonense (100 mL ha-1), 2) biological fertilizer without the addition of N. amazonense, 3) inoculation with N. amazonense at a dose of 100 mL ha-1 with 2x108 viable cell mL-1, 4) inoculation with N. amazonense at a dose of 200 mL ha-1 with 2x108 viable cell mL-1, and 5) control, without the addition of biological fertilizer and N. amazonense. The biological fertilizer was applied at dose of 300 L ha-1, which was split at planting (200 L ha-1) and in the post-emergence phase (100 L ha-1). After 164 days of planting, it was detected an increase in leaf length +3, number of green leaves, leaf area and sulfur content in the leaves of sugarcane plants that received the biological fertilizer with the addition of N. amazonense. In conclusion, the combination of N. amazonense with biological fertilizer revealed positive effects through morphological and nutritional characteristics in sugarcane plants during their early stages of development when compared to plants grown only with the inoculation of N. amazonense, biological fertilizer or mineral fertilizers, with few notable positive effects on soil chemistry.

Neste estudo avaliou-se a hipótese de que a combinação de Nitrospirillum amazonense estirpe BR11145 com adubo biológico preparado com base em microbioma autóctone da localidade de aplicação do produto, produzido com Microgeo®, resulta em benefícios morfológicos e nutricionais para o desenvolvimento inicial de plantas de cana-de-açúcar e para a química do solo. Para tanto, mudas pré-brotadas de cana-de-açúcar foram crescidas em mesocosmos num experimento conduzido em casa-de-vegetação com solo enriquecido com fertilizante à base de nitrogênio/fósforo/potássio. Os tratamentos avaliados foram: 1) adubo biológico acrescido de N. amazonense (100 mL ha-1), 2) adubo biológico sem o acréscimo de N. amazonense, 3) inoculação de N. amazonense na dose de 100 mL ha-1 (2x108 células viáveis por mL), 4) inoculação de N. amazonense na dose de 200 mL ha-1 (2x108 células viáveis por mL), e 5) testemunha, sem a adição de fertilizante biológico e N. amazonense. O adubo biológico foi aplicado na dose de 300 L ha-1 sendo esta parcelada no plantio (200 L ha-1) e na fase de pós-emergência (100 L ha-1). Após 164 dias do plantio constatou-se aumento no comprimento da folha +3, número de folhas verdes, área foliar e no teor de enxofre nas folhas das plantas de cana-de-açúcar que receberam o adubo biológico combinado com N. amazonense. Conclui-se que a combinação de N. amazonense com o adubo biológico produzido com Microgeo® revelou efeitos positivos por meio de caraterísticas morfológicas e nutricionais em plantas de cana-de-açúcar durante os seus estádios iniciais de desenvolvimento quando comparadas com plantas crescidas apenas com a inoculação de N. amazonense, fertilizante biológico ou fertilizantes minerais, com poucos efeitos positivos notáveis na química do solo.

Enhancing Phosphorus and Nitrogen Uptake in Maize Crops with Food Industry Biosolids and Azotobacter nigricans.

The problem of phosphorus and nitrogen deficiency in agricultural soils has been solved by adding chemical fertilizers. However, their excessive use and their accumulation have only contributed to environmental contamination. Given the high content of nutrients in biosolids collected from a food industry waste treatment plant, their use as fertilizers was investigated in Zea mays plants grown in sandy loam soil collected from a semi-desert area. These biosolids contained insoluble phosphorus sources; therefore, given the ability of Azotobacter nigricans to solubilize phosphates, this strain was incorporated into the study. In vitro, the suitable conditions for the growth of Z. mays plants were determined by using biosolids as a fertilizer and A. nigricans as a plant-growth-promoting microorganism; in vitro, the ability of A. nigricans to solubilize phosphates, fix nitrogen, and produce indole acetic acid, a phytohormone that promotes root formation, was also evaluated. At the greenhouse stage, the Z. mays plants fertilized with biosolids at concentrations of 15 and 20% (v/w) and inoculated with A. nigricans favored the development of bending strength plants, which was observed on the increased stem diameter (>13.5% compared with the negative control and >7.4% compared with the positive control), as well as a better absorption of phosphorus and nitrogen, the concentration of which increased up to 62.8% when compared with that in the control treatments. The interactions between plants and A. nigricans were observed via scanning electron microscopy. The application of biosolids and A. nigricans in Z. mays plants grown in greenhouses presented better development than when Z. mays plants were treated with a chemical fertilizer. The enhanced plant growth was attributed to the increase in root surface area.

The Friend Within: Endophytic Bacteria as a Tool for Sustainability in Strawberry Crops.

Strawberry (Fragaria x ananassa, Duch.) is an important crop worldwide. However, since it is a highly demanding crop in terms of the chemical conditions of the substrate, a large part of strawberry production implies the application of large amounts of fertilizers in the production fields. This practice can cause environmental problems, in addition to increases in the fruit's production costs. In this context, applying plant growth-promoting bacteria in production fields can be an essential strategy, especially thanks to their ability to stimulate plant growth via different mechanisms. Therefore, this study aimed to test in vitro and in vivo the potential of bacteria isolated from strawberry leaves and roots to directly promote plant growth. The isolates were tested in vitro for their ability to produce auxins, solubilize phosphate and fix nitrogen. Isolates selected in vitro were tested on strawberry plants to promote plant growth and increase the accumulation of nitrogen and phosphorus in the leaves. The tested isolates showed an effect on plant growth according to biometric parameters. Among the tested isolates, more expressive results for the studied variables were observed with the inoculation of the isolate MET12M2, belonging to the species Brevibacillus fluminis. In general, bacterial inoculation induced strain-dependent effects on strawberry growth. In vitro and in vivo assays showed the potential use of the B. fluminis MET12M2 isolate as a growth promoter for strawberries.

Gram-negative bacteria associated with a dominant arboreal ant species outcompete phyllosphere-associated bacteria species in a tropical canopy.

Ants have efficient and well-studied social immunity mechanisms, which prevent the colony contamination. Little is known about how workers keep their outside territory clear of diseases. We investigated the interactions between Azteca chartifex ants, their associated bacteria and bacteria on the phyllosphere of Byrsonima sericea trees, comparing plants patrolled and not by the ants. The hypothesis is that bacteria associated with the worker's exoskeleton may outcompete the leaf bacteria. Culturable bacteria were isolated from ants, from the main and satellite nests, and from phyllosphere of B. sericea taken from trees that had A. chartifex nests and from trees without nests. The isolates were grouped by Gram guilds and identified at the genus level. There was a higher percentage of Gram-negative isolates in the ants and on the leaves patrolled by them. There was a higher growth rate of ant bacteria from the main nest compared to those found in ants from the satellite nests. The most representative genus among ant isolates was Enterobacter, also found on leaves patrolled by ants. Under favourable in vitro conditions, A. chartifex Gram-negative bacteria outcompete leaf bacteria by overgrowth, showing a greater competition capacity over the Gram-positive bacteria from leaves with no previous interaction with ants in the field. It was demonstrated that ants carry bacteria capable of outcompeting exogenous bacteria associated with their outside territory. The leaf microbiota of a patrolled tree could be shaped by the ant microbiota, suggesting that large ant colonies may have a key role in structuring canopy plant-microbe interactions.

Growth promotion in Arabidopsis thaliana by bacterial cyclodipeptides involves the TOR/S6K pathway activation.

Cyclodipeptides (CDPs) are the smallest peptidic molecules that can be produced by diverse organisms such as bacteria, fungi, and animals. They have multiple biological effects. In this paper, we examined the CDPs produced by the bacteria Pseudomonas aeruginosa PAO1, which are known as opportunistic pathogens of humans and plants on TARGET OF RAPAMYCIN (TOR) signaling pathways, and regulation of root system architecture. This bacterium produces the bioactive CDPs: cyclo(L-Pro-L-Leu), cyclo(L-Pro-L-Phe), cyclo(L-Pro-L-Tyr), and cyclo(L-Pro-L-Val). In a previous report, these molecules were found to modulate basic cellular programs not only via auxin mechanisms but also by promoting the phosphorylation of the S6 ribosomal protein kinase (S6K), a downstream substrate of the TOR kinase. In the present work, we found that the inoculation of Arabidopsis plants with P. aeruginosa PAO1, the non-pathogenic P. aeruginosa ΔlasI/Δrhll strain (JM2), or by direct exposure of plants to CDPs influenced growth and promoted root branching depending upon the treatment imposed, while genetic evidence using Arabidopsis lines with enhanced or decreased TOR levels indicated a critical role of this pathway in the bacterial phytostimulation.

Insights into the early stages of plant-endophytic bacteria interaction.

The plant holobiont is a complex entity composed of the plant and the organisms that live in and on it including its microbiota. The plant microbiota includes, among other microorganisms, bacterial endophytes, which are bacteria that can invade living plant tissues without causing symptoms of disease. The interaction between the endophytic bacterial microbiota and their plant host has profound influences on their fitness and depends on biotic and abiotic factors. For these interactions to be established, the bacteria have to be present at the right time, in the right place either colonizing the soil or the seed. In this review we summarize the current knowledge regarding the sources of the bacterial endophytic microbiome and the processes involved in the assemblage of the resulting community during the initial stages of plant development. The adaptations that allow the spatial approximation of soil- and seed-borne bacteria towards infection and colonization of the internal tissues of plants will be addressed in this review.

The protective role of PHB and its degradation products against stress situations in bacteria.

Many bacteria produce storage biopolymers that are mobilized under conditions of metabolic adaptation, for example, low nutrient availability and cellular stress. Polyhydroxyalkanoates are often found as carbon storage in Bacteria or Archaea, and of these polyhydroxybutyrate (PHB) is the most frequently occurring PHA type. Bacteria usually produce PHB upon availability of a carbon source and limitation of another essential nutrient. Therefore, it is widely believed that the function of PHB is to serve as a mobilizable carbon repository when bacteria face carbon limitation, supporting their survival. However, recent findings indicate that bacteria switch from PHB synthesis to mobilization under stress conditions such as thermal and oxidative shock. The mobilization products, 3-hydroxybutyrate and its oligomers, show a protective effect against protein aggregation and cellular damage caused by reactive oxygen species and heat shock. Thus, bacteria should have an environmental monitoring mechanism directly connected to the regulation of the PHB metabolism. Here, we review the current knowledge on PHB physiology together with a summary of recent findings on novel functions of PHB in stress resistance. Potential applications of these new functions are also presented.

Membrane Homeoviscous Adaptation in Sinorhizobium Submitted to a Stressful Thermal Cycle Contributes to the Maintenance of the Symbiotic Plant-Bacteria Interaction.

Here, we estimate fast changes in the fluidity of Sinorhizobium meliloti membranes submitted to cyclic temperature changes (10°C-40°C-10°C) by monitoring the fluorescence polarization (P) of DPH and TMA-DPH of the whole cell (WC) as well as in its outer (OM) and inner (IM) membranes. Additionally, the long-term response to thermal changes is demonstrated through the dynamics of the phospholipid and fatty acid composition in each membrane. This allowed membrane homeoviscous adaptation by the return to optimal fluidity levels as measured by the PDPH/TMA-DPH in WC, OM, IM, and multilamellar vesicles of lipids extracted from OM and IM. Due to probe-partitioning preferences and membranes' compositional characteristics, DPH and TMA-DPH exhibit different behaviors in IM and OM. The rapid effect of cyclic temperature changes on the P was the opposite in both membranes with the IM being the one that exhibited the thermal behavior expected for lipid bilayers. Interestingly, only after the incubation at 40°C, cells were unable to recover the membrane preheating P levels when cooled up to 10°C. Solely in this condition, the formation of threads and nodular structures in Medicago sativa infected with S. meliloti were delayed, indicating that the symbiotic interaction was partially altered but not halted.

Interaction among endophytic bacteria, sweet sorghum (Sorghum bicolor) cultivars and chemical nitrogen fertilization.

The application of new agricultural technologies to attain sustainable production systems is necessary. The use of plant growth-promoting bacteria to improve plant growth and health has been studied for decades. This work aimed to isolate diazotrophic endophytic bacteria associated with sweet sorghum plants and study the interaction of their inoculation in combination with chemical N-fertilization on different sorghum cultivars. A bacterial collection of 181 isolates was constructed and characterized in vitro and in vivo. From that, the strains Enterobacter sp. UYSB89 and Kosakonia sp. UYSB139 were nifH+, produce IAA, defined as true endophytes and able to promote growth of two sweet sorghum under greenhouse conditions. The evaluated cultivars responded differentially to bacterial inoculation, the nitrogen fertilization doses and their interaction. Thus, plant growth is a multifactorial consequence of the interrelation between crop practices and the plant genotypes. This knowledge is a valuable factor in terms of understanding plant-bacteria endophyte interactions to preserve environmental sustainability during the implementation of agronomic practices.

Herbaspirillum rubrisubalbicans as a Phytopathogenic Model to Study the Immune System of Sorghum bicolor.

Herbaspirillum rubrisubalbicans is the causal agent of red stripe disease (RSD) and mottle stripe disease of sorghum and sugarcane, respectively. In all, 63 genotypes of Sorghum bicolor were inoculated with H. rubrisubalbicans, with 59 showing RSD symptoms. Quantitative trait loci (QTL) analysis in a recombinant inbred line (RIL) population identified several QTL associated with variation in resistance to RSD. RNA sequencing analysis identified a number of genes whose transcript levels were differentially regulated during H. rubrisubalbicans infection. Among those genes that responded to H. rubrisubalbicans inoculation were many involved in plant-pathogen interactions such as leucine-rich repeat receptors, mitogen-activated protein kinase 1, calcium-binding proteins, transcriptional factors (ethylene-responsive element binding factor), and callose synthase. Pretreatment of sorghum leaves with the pathogen-associated molecular pattern (PAMP) molecules flg22 and chitooctaose provided protection against subsequent challenge with the pathogen, suggesting that PAMP-triggered immunity plays an important role in the sorghum immunity response. These data present baseline information for the use of the genetically tractable H. rubrisubalbicans-sorghum pathosystem for the study of innate immunity and disease resistance in this important grain and bioenergy crop. Information gained from the use of this system is likely to be informative for other monocots, including those more intractable for experimental study (e.g., sugarcane).

Isolation, Biochemical and Genomic Characterization of Glyphosate Tolerant Bacteria to Perform Microbe-Assisted Phytoremediation.

The large-scale use of the herbicide glyphosate leads to growing ecotoxicological and human health concerns. Microbe-assisted phytoremediation arises as a good option to remove, contain, or degrade glyphosate from soils and waterbodies, and thus avoid further spreading to non-target areas. To achieve this, availability of plant-colonizing, glyphosate-tolerant and -degrading strains is required and at the same time, it must be linked to plant-microorganism interaction studies focusing on a substantive ability to colonize the roots and degrade or transform the herbicide. In this work, we isolated bacteria from a chronically glyphosate-exposed site in Argentina, evaluated their glyphosate tolerance using the minimum inhibitory concentration assay, their in vitro degradation potential, their plant growth-promotion traits, and performed whole genome sequencing to gain insight into the application of a phytoremediation strategy to remediate glyphosate contaminated agronomic soils. Twenty-four soil and root-associated bacterial strains were isolated. Sixteen could grow using glyphosate as the sole source of phosphorous. As shown in MIC assay, some strains tolerated up to 10000 mg kg-1 of glyphosate. Most of them also demonstrated a diverse spectrum of in vitro plant growth-promotion traits, confirmed in their genome sequences. Two representative isolates were studied for their root colonization. An isolate of Ochrobactrum haematophilum exhibited different colonization patterns in the rhizoplane compared to an isolate of Rhizobium sp. Both strains were able to metabolize almost 50% of the original glyphosate concentration of 50 mg l-1 in 9 days. In a microcosms experiment with Lotus corniculatus L, O. haematophilum performed better than Rhizobium, with 97% of glyphosate transformed after 20 days. The results suggest that L. corniculatus in combination with to O. haematophilum can be adopted for phytoremediation of glyphosate on agricultural soils. An effective strategy is presented of linking the experimental data from the isolation of tolerant bacteria with performing plant-bacteria interaction tests to demonstrate positive effects on the removal of glyphosate from soils.

Interaction between sugarcane families and plant growth-promoting bacteria in two crop cycles / Interação entre famílias de cana-de-açúcar e bactérias promotoras do crescimento vegetal em dois ciclos de cultivo

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)

Interaction between sugarcane families and plant growth-promoting bacteria in two crop cycles / Interação entre famílias de cana-de-açúcar e bactérias promotoras do crescimento vegetal em dois ciclos de cultivo

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...

Enhanced drought tolerance in seedlings of Neotropical tree species inoculated with plant growth-promoting bacteria.

The inoculation of tree species with plant growth-promoting bacteria (PGPB) has emerged as an important strategy for the acclimation of seedlings by improving plant tolerance to biotic and abiotic stresses. This study aimed to evaluate the effects of inoculation with bacterial species (Azospirillum brasilense - Ab-V5, Bacillus sp., Azomonas sp. and Azorhizophillus sp.) on the growth and physiology of the Neotropical tree species Trema micrantha and Cariniana estrellensis under drought conditions. When associated with Ab-V5 and Azomonas sp., T. micrantha showed increased protein in the leaves, starch in the leaves and roots, photosynthesis, instantaneous carboxylation efficiency and root and shoot dry mass. Moreover, there were reductions in hydrogen peroxide, lipid peroxidation, water potential and proline. In C. estrellensis associated with Ab-V5, higher values of photosynthesis and instantaneous carboxylation efficiency were observed, in addition to higher starch content in the leaves and roots and higher protein content in the leaves; lower hydrogen peroxide and lipid peroxidation contents were also observed. The associations of T. micrantha with Ab-V5 and Azomonas sp. and C. estrellensis with Ab-V5 favored the activation of metabolic processes under drought, leading to greater drought tolerance. This work demonstrates the effects of compatible associations of Neotropical tree and PGPB species and suggests that the identification of compatible PGPB strains can result in tree seedlings with increased tolerance to abiotic stresses, such as drought.

Genome wide transcriptional profiling of Herbaspirillum seropedicae SmR1 grown in the presence of naringenin.

Herbaspirillum seropedicae is a diazotrophic bacterium which associates endophytically with economically important gramineae. Flavonoids such as naringenin have been shown to have an effect on the interaction between H. seropedicae and its host plants. We used a high-throughput sequencing based method (RNA-Seq) to access the influence of naringenin on the whole transcriptome profile of H. seropedicae. Three hundred and four genes were downregulated and seventy seven were upregulated by naringenin. Data analysis revealed that genes related to bacterial flagella biosynthesis, chemotaxis and biosynthesis of peptidoglycan were repressed by naringenin. Moreover, genes involved in aromatic metabolism and multidrug transport efllux were actived.

Methylobacterium-plant interaction genes regulated by plant exudate and quorum sensing molecules

Bacteria from the genus Methylobacterium interact symbiotically (endophytically and epiphytically) with different plant species. These interactions can promote plant growth or induce systemic resistance, increasing plant fitness. The plant colonization is guided by molecular communication between bacteria-bacteria and bacteria-plants, where the bacteria recognize specific exuded compounds by other bacteria (e.g. homoserine molecules) and/or by the plant roots (e.g. flavonoids, ethanol and methanol), respectively. In this context, the aim of this study was to evaluate the effect of quorum sensing molecules (N-acyl-homoserine lactones) and plant exudates (including ethanol) in the expression of a series of bacterial genes involved in Methylobacterium-plant interaction. The selected genes are related to bacterial metabolism (mxaF), adaptation to stressful environment (crtI, phoU and sss), to interactions with plant metabolism compounds (acdS) and pathogenicity (patatin and phoU). Under in vitro conditions, our results showed the differential expression of some important genes related to metabolism, stress and pathogenesis, thereby AHL molecules up-regulate all tested genes, except phoU, while plant exudates induce only mxaF gene expression. In the presence of plant exudates there is a lower bacterial density (due the endophytic and epiphytic colonization), which produce less AHL, leading to down regulation of genes when compared to the control. Therefore, bacterial density, more than plant exudate, influences the expression of genes related to plant-bacteria interaction.

Methylobacterium-plant interaction genes regulated by plant exudate and quorum sensing molecules.

Bacteria from the genus Methylobacterium interact symbiotically (endophytically and epiphytically) with different plant species. These interactions can promote plant growth or induce systemic resistance, increasing plant fitness. The plant colonization is guided by molecular communication between bacteria-bacteria and bacteria-plants, where the bacteria recognize specific exuded compounds by other bacteria (e.g. homoserine molecules) and/or by the plant roots (e.g. flavonoids, ethanol and methanol), respectively. In this context, the aim of this study was to evaluate the effect of quorum sensing molecules (N-acyl-homoserine lactones) and plant exudates (including ethanol) in the expression of a series of bacterial genes involved in Methylobacterium-plant interaction. The selected genes are related to bacterial metabolism (mxaF), adaptation to stressful environment (crtI, phoU and sss), to interactions with plant metabolism compounds (acdS) and pathogenicity (patatin and phoU). Under in vitro conditions, our results showed the differential expression of some important genes related to metabolism, stress and pathogenesis, thereby AHL molecules up-regulate all tested genes, except phoU, while plant exudates induce only mxaF gene expression. In the presence of plant exudates there is a lower bacterial density (due the endophytic and epiphytic colonization), which produce less AHL, leading to down regulation of genes when compared to the control. Therefore, bacterial density, more than plant exudate, influences the expression of genes related to plant-bacteria interaction.

Methylobacterium-plant interaction genes regulated by plant exudate and quorum sensing molecules

Bacteria from the genus Methylobacterium interact symbiotically (endophytically and epiphytically) with different plant species. These interactions can promote plant growth or induce systemic resistance, increasing plant fitness. The plant colonization is guided by molecular communication between bacteria-bacteria and bacteria-plants, where the bacteria recognize specific exuded compounds by other bacteria (e.g. homoserine molecules) and/or by the plant roots (e.g. flavonoids, ethanol and methanol), respectively. In this context, the aim of this study was to evaluate the effect of quorum sensing molecules (N-acyl-homoserine lactones) and plant exudates (including ethanol) in the expression of a series of bacterial genes involved in Methylobacterium-plant interaction. The selected genes are related to bacterial metabolism (mxaF), adaptation to stressful environment (crtI, phoU and sss), to interactions with plant metabolism compounds (acdS) and pathogenicity (patatin and phoU). Under in vitro conditions, our results showed the differential expression of some important genes related to metabolism, stress and pathogenesis, thereby AHL molecules up-regulate all tested genes, except phoU, while plant exudates induce only mxaF gene expression. In the presence of plant exudates there is a lower bacterial density (due the endophytic and epiphytic colonization), which produce less AHL, leading to down regulation of genes when compared to the control. Therefore, bacterial density, more than plant exudate, influences the expression of genes related to plant-bacteria interaction.

Analysis of 16S rRNA and mxaF genes revealing insights into Methylobacterium niche-specific plant association.

The genus Methylobacterium comprises pink-pigmented facultative methylotrophic (PPFM) bacteria, known to be an important plant-associated bacterial group. Species of this group, described as plant-nodulating, have the dual capacity of producing cytokinin and enzymes, such as pectinase and cellulase, involved in systemic resistance induction and nitrogen fixation under specific plant environmental conditions. The aim hereby was to evaluate the phylogenetic distribution of Methylobacterium spp. isolates from different host plants. Thus, a comparative analysis between sequences from structural (16S rRNA) and functional mxaF (which codifies for a subunit of the enzyme methanol dehydrogenase) ubiquitous genes, was undertaken. Notably, some Methylobacterium spp. isolates are generalists through colonizing more than one host plant, whereas others are exclusively found in certain specific plant-species. Congruency between phylogeny and specific host inhabitance was higher in the mxaF gene than in the 16S rRNA, a possible indication of function-based selection in this niche. Therefore, in a first stage, plant colonization by Methylobacterium spp. could represent generalist behavior, possibly related to microbial competition and adaptation to a plant environment. Otherwise, niche-specific colonization is apparently impelled by the host plant.

Analysis of 16S rRNA and mxaF genes reveling insights into Methylobacterium niche-specific plant association

The genus Methylobacterium comprises pink-pigmented facultative methylotrophic (PPFM) bacteria, known to be an important plant-associated bacterial group. Species of this group, described as plant-nodulating, have the dual capacity of producing cytokinin and enzymes, such as pectinase and cellulase, involved in systemic resistance induction and nitrogen fixation under specific plant environmental conditions. The aim hereby was to evaluate the phylogenetic distribution of Methylobacterium spp. isolates from different host plants. Thus, a comparative analysis between sequences from structural (16S rRNA) and functional mxaF (which codifies for a subunit of the enzyme methanol dehydrogenase) ubiquitous genes, was undertaken. Notably, some Methylobacterium spp. isolates are generalists through colonizing more than one host plant, whereas others are exclusively found in certain specific plant-species. Congruency between phylogeny and specific host inhabitance was higher in the mxaF gene than in the 16S rRNA, a possible indication of function-based selection in this niche. Therefore, in a first stage, plant colonization by Methylobacterium spp. could represent generalist behavior, possibly related to microbial competition and adaptation to a plant environment. Otherwise, niche-specific colonization is apparently impelled by the host plant.