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1.
Biomolecules ; 10(4)2020 04 16.
Article in English | MEDLINE | ID: mdl-32316239

ABSTRACT

Motile Methylobacterium sp. ME121 and non-motile Kaistia sp. 32K were isolated from the same soil sample. Interestingly, ME121 was significantly more motile in the coculture of ME121 and 32K than in the monoculture of ME121. This advanced motility of ME121 was also observed in the 32K culture supernatant. A swimming acceleration factor, which we named the K factor, was identified in the 32K culture supernatant, purified, characterized as an extracellular polysaccharide (5-10 kDa), and precipitated with 70% ethanol. These results suggest the possibility that the K factor was directly or indirectly sensed by the flagellar stator, accelerating the flagellar rotation of ME121. To the best of our knowledge, no reports describing an acceleration in motility due to coculture with two or more types of bacteria have been published. We propose a mechanism by which the increase in rotational force of the ME121 flagellar motor is caused by the introduction of the additional stator into the motor by the K factor.


Subject(s)
Bacterial Proteins/metabolism , Methylobacterium/physiology , Rhizobiaceae/metabolism , Chemical Precipitation , Ethanol/chemistry , Flagella/metabolism , Methylobacterium/growth & development , Monosaccharides/analysis , Movement , Rotation
2.
mSphere ; 5(1)2020 01 15.
Article in English | MEDLINE | ID: mdl-31941811

ABSTRACT

Microbial communities in the evaporator core (EC) of automobile air-conditioning systems have a large impact on indoor air quality, such as malodor and allergenicity. DNA-based microbial population analysis of the ECs collected from South Korea, China, the United States, India, and the United Arab Emirates revealed the extraordinary dominance of Methylobacterium species in EC biofilms. Mixed-volatile organic compound (VOC) utilization and biofilm-forming capabilities were evaluated to explain the dominance of Methylobacterium species in the ECs. The superior growth of all Methylobacterium species could be possible under mixed-VOC conditions. Interestingly, two lifestyle groups of Methylobacterium species could be categorized as the aggregator group, which sticks together but forms a small amount of biofilm, and the biofilm-forming group, which forms a large amount of biofilm, and their genomes along with phenotypic assays were analyzed. Pili are some of the major contributors to the aggregator lifestyle, and succinoglycan exopolysaccharide production may be responsible for the biofilm formation. However, the coexistence of these two lifestyle Methylobacterium groups enhanced their biofilm formation compared to that with each single culture.IMPORTANCE Air-conditioning systems (ACS) are indispensable for human daily life; however, microbial community analysis in automobile ACS has yet to be comprehensively investigated. A bacterial community analysis of 24 heat exchanger fins from five countries (South Korea, China, the United States, India, and the United Arab Emirates [UAE]) revealed that Methylobacterium species are some of the dominant bacteria in automobile ACS. Furthermore, we suggested that the predominance of Methylobacterium species in automobile ACS is due to the utilization of mixed volatile organic compounds and their great ability for aggregation and biofilm formation.


Subject(s)
Air Conditioning , Automobiles , Biofilms/growth & development , Methylobacterium/physiology , Microbiota , China , India , Republic of Korea , United Arab Emirates , United States , Volatile Organic Compounds/metabolism
3.
Microbes Environ ; 35(1)2020.
Article in English | MEDLINE | ID: mdl-31969531

ABSTRACT

The Vietnamese Mekong delta is one of the largest rice-producing areas globally. Methylobacterium spp. are persistent colonizers of the rice plant and exert beneficial effects on plant growth and health. Sixty-one Methylobacterium strains belonging to seven species were predominantly isolated from the phyllosphere of rice cultivated in six Mekong delta provinces. Inoculation tests revealed that some strains exhibited plant growth-promoting activity. Moreover, three strains possessed the novel characteristics of inducing leaf bleaching and killing rice seedlings. These results revealed the complex diversity of Methylobacterium in Mekong delta rice and that healthy and productive rice cultivation requires a proper balance of Methylobacterium.


Subject(s)
Genetic Variation , Methylobacterium/genetics , Oryza/microbiology , Lactuca/growth & development , Lactuca/microbiology , Methylobacterium/classification , Methylobacterium/physiology , Oryza/growth & development , Plant Components, Aerial/microbiology , Plant Growth Regulators/physiology , Vietnam
4.
FEMS Microbiol Ecol ; 95(12)2019 12 01.
Article in English | MEDLINE | ID: mdl-31697309

ABSTRACT

Greenland's Dark Zone is the largest contiguous region of bare terrestrial ice in the Northern Hemisphere and microbial processes play an important role in driving its darkening and thereby amplifying melt and runoff from the ice sheet. However, the dynamics of these microbiota have not been fully identified. Here, we present joint 16S rRNA gene and 16S rRNA (cDNA) comparison of input (snow), storage (cryoconite) and output (supraglacial stream water) habitats across the Dark Zone over the melt season. We reveal that all three Dark Zone communities have a preponderance of rare taxa exhibiting high protein synthesis potential (PSP). Furthermore, taxa with high PSP represent highly connected 'bottlenecks' within community structure, consistent with their roles as metabolic hubs. Finally, low abundance-high PSP taxa affiliated with Methylobacterium within snow and stream water suggest a novel role for Methylobacterium in the carbon cycle of Greenlandic snowpacks, and importantly, the export of potentially active methylotrophs to the bed of the Greenland Ice Sheet. By comparing the dynamics of bulk and potentially active microbiota in the Dark Zone of the Greenland Ice Sheet, we provide novel insights into the mechanisms and impacts of the microbial colonization of this critical region of our melting planet.


Subject(s)
Carbon Cycle/physiology , Ice Cover/microbiology , Methylobacterium/physiology , Snow/microbiology , Ecosystem , Freezing , Greenland , Microbiota/physiology , RNA, Ribosomal, 16S/genetics , Seasons
5.
Planta ; 249(6): 1903-1919, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30877435

ABSTRACT

MAIN CONCLUSION: Inoculation of endophytic Methylobacterium oryzae CBMB20 in salt-stressed rice plants improves photosynthesis and reduces stress volatile emissions due to mellowing of ethylene-dependent responses and activating vacuolar H+-ATPase. The objective of this study was to analyze the impact of ACC (1-aminocyclopropane-1-carboxylate) deaminase-producing Methylobacterium oryzae CBMB20 in acclimation of plant to salt stress by controlling photosynthetic characteristics and volatile emission in salt-sensitive (IR29) and moderately salt-resistant (FL478) rice (Oryza sativa L.) cultivars. Saline levels of 50 mM and 100 mM NaCl with and without bacteria inoculation were applied, and the temporal changes in stress response and salinity resistance were assessed by monitoring photosynthetic characteristics, ACC accumulation, ACC oxidase activity (ACO), vacuolar H+ ATPase activity, and volatile organic compound (VOC) emissions. Salt stress considerably reduced photosynthetic rate, stomatal conductance, PSII efficiency and vacuolar H+ ATPase activity, but it increased ACC accumulation, ACO activity, green leaf volatiles, mono- and sesquiterpenes, and other stress volatiles. These responses were enhanced with increasing salt stress and time. However, rice cultivars treated with CBMB20 showed improved plant vacuolar H+ ATPase activity, photosynthetic characteristics and decreased ACC accumulation, ACO activity and VOC emission. The bacteria-dependent changes were greater in the IR29 cultivar. These results indicate that decreasing photosynthesis and vacuolar H+ ATPase activity rates and increasing VOC emission rates in response to high-salinity stress were effectively mitigated by M. oryzae CBMB20 inoculation.


Subject(s)
Ethylenes/metabolism , Methylobacterium/physiology , Oryza/microbiology , Photosynthesis , Plant Growth Regulators/metabolism , Volatile Organic Compounds/metabolism , Amino Acid Oxidoreductases/genetics , Amino Acid Oxidoreductases/metabolism , Carbon-Carbon Lyases/genetics , Carbon-Carbon Lyases/metabolism , Endophytes , Genotype , Oryza/enzymology , Oryza/genetics , Oryza/physiology , Phenotype , Plant Proteins/genetics , Plant Proteins/metabolism , Salinity , Salt Stress , Stress, Physiological , Vacuolar Proton-Translocating ATPases/genetics , Vacuolar Proton-Translocating ATPases/metabolism
6.
Biosci Biotechnol Biochem ; 83(3): 569-577, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30475153

ABSTRACT

A number of pink-pigmented facultative methylotrophs (PPFMs) belonging to Methylobacterium spp. isolated from living plant samples were found to require B vitamins for their growth in minimal medium, and most B vitamin-auxotrophic PPFMs required pantothenate (vitamin B5). Further investigation of pantothenate auxotrophy using the representative strain Methylobacterium sp. OR01 demonstrated that this strain cannot synthesize ß-alanine, one of the precursors of pantothenate. ß-alanine and several precursors of pantothenate restored the growth of Methylobacterium sp. OR01 in minimal medium. Furthermore, this strain could colonize leaves of Arabidopsis thaliana cultivated in medium without pantothenate or its precursors. Pantothenate, ß-alanine and several precursors were detected in the suspension of A. thaliana leaves. These results suggest that pantothenate-auxotrophic PPFMs can symbiotically colonize the surface of plant leaves by acquiring ß-alanine and other precursors, in addition to pantothenate. Finally, the fitness advantage of B vitamin auxotrophy of PPFMs in the phyllosphere environment is discussed.


Subject(s)
Autotrophic Processes , Methylobacterium/metabolism , Pantothenic Acid/metabolism , Arabidopsis/microbiology , Methylobacterium/physiology , Plant Leaves/microbiology , beta-Alanine/metabolism
7.
Physiol Plant ; 166(3): 729-747, 2019 Jul.
Article in English | MEDLINE | ID: mdl-30175853

ABSTRACT

The study was envisaged to assess the extent of normally uncultivable endophytic bacteria in field papaya plants and in vitro established cultures adopting cultivation vs molecular analysis and microscopy. Surface-sterilized axillary shoot-buds of papaya 'Arka Surya' revealed high bacterial diversity as per 16S rRNA metagene amplicon sequencing (6 phyla, 10 classes, 21 families) with an abundance of Pseudomonas (Gammaproteobacteria), which also formed a common contaminant for in vitro cultured field explants. Molecular analysis of seedling shoot-tip-derived healthy proliferating cultures of three genotypes ('Arka Surya', 'Arka Prabhath', 'Red Lady') with regular monthly subculturing also displayed high bacterial diversity (11-16 phyla, >25 classes, >50 families, >200 genera) about 12-18 months after initial establishment. 'Arka Surya' and 'Red Lady' cultures bore predominantly Actinobacteria (75-78%) while 'Arka Prabhath' showed largely Alphaproteobacteria corroborating the slowly activated Methylobacterium sp. Bright-field direct microscopy on tissue sections and tissue homogenate and epi-fluorescence microscopy employing bacterial DNA probe SYTO-9 revealed abundant intracellular bacteria embracing the next-generation sequencing elucidated high taxonomic diversity. Phylogenetic investigation of communities by reconstruction of unobserved states- PICRUSt- functional annotation suggested significant operational roles for the bacterial-biome. Metabolism, environmental information processing, and genetic information processing constituted major Kyoto Encyclopedia of Genes and Genomes KEGG attributes. Papaya stocks occasionally displayed bacterial growth on culture medium arising from the activation of originally uncultivable organisms to cultivation. The organisms included Bacillus (35%), Methylobacterium (15%), Pseudomonas (10%) and seven other genera (40%). This study reveals a hidden world of diverse and abundant conventionally uncultivable cellular-colonizing endophytic bacteria in field shoots and micropropagating papaya stocks with high genotypic similarity and silent participation in various plant processes/pathways.


Subject(s)
Carica/microbiology , Endophytes/physiology , Plant Shoots/microbiology , Alphaproteobacteria/genetics , Alphaproteobacteria/physiology , Bacillus/genetics , Bacillus/physiology , Endophytes/genetics , Genotype , Methylobacterium/genetics , Methylobacterium/physiology , Phylogeny , Plant Shoots/genetics , Pseudomonas/genetics , Pseudomonas/physiology , RNA, Ribosomal, 16S/genetics
8.
Biosci Biotechnol Biochem ; 83(1): 174-180, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30200829

ABSTRACT

Numerous gram-negative bacteria have quorum-sensing systems and produce AHL as a quorum-sensing signal molecule. In this study, we demonstrated that Methylobacterium populi P-1M, an isolate from a pink-pigmented household biofilm, produced two AHLs, C14:1-HSL as a predominant product and 3OHC14-HSL as a minor product. The complete genome sequence of M. populi P-1M revealed the presence of genes that are predicted to encode an AHL synthase (mpoI) and AHL receptor (mpoR). M. populi P-1M formed a pellicle-like biofilm, which had a flat surface and was easily removable. In contrast, biofilms formed by mpoI and/or mpoR deletion mutants had a wavy surface structure and strongly adhered to the glass tube. When C14:1-HSL was added to the mpoI mutant culture, the biofilm structure resembled that of the wild-type strain. These results demonstrated that the structure and adhesion strength of M. populi P-1M biofilms are determined in part by AHL-mediated quorum sensing.Abbreviations: AHL: N-acyl-l-homoserine lactone; C14:1-HSL: N-tetradecenoyl-l-homoserine lactone; 3OHC14-HSL: N-(3-hydroxytetradecanoyl)-l-homoserine lactone; SAM: S-adenosyl-l-methionine; ACP: acyl-acyl carrier protein; EPS: extracellular polysaccharide; DMSO: dimethyl sulfoxide.


Subject(s)
4-Butyrolactone/analogs & derivatives , Biofilms/growth & development , Housing , Methylobacterium/cytology , Methylobacterium/physiology , Pigmentation , Quorum Sensing , 4-Butyrolactone/metabolism , Methylobacterium/genetics , Methylobacterium/metabolism , Mutation
10.
Protoplasma ; 254(5): 1867-1877, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28054231

ABSTRACT

In numerous experimental studies, seedlings of the model dicot Arabidopsis thaliana have been raised on sterile mineral salt agar. However, under natural conditions, no plant has ever grown in an environment without bacteria. Here, we document that germ-free (gnotobiotic) seedlings, raised on mineral salt agar without sucrose, develop very short root hairs. In the presence of a soil extract that contains naturally occurring microbes, root hair elongation is promoted; this effect can be mimicked by the addition of methylobacteria to germ-free seedlings. Using five different bacterial species (Methylobacterium mesophilicum, Methylobacterium extorquens, Methylobacterium oryzae, Methylobacterium podarium, and Methylobacterium radiotolerans), we show that, over 9 days of seedling development in a light-dark cycle, root development (hair elongation, length of the primary root, branching patterns) is regulated by these epiphytic microbes that occur in the rhizosphere of field-grown plants. In a sterile liquid culture test system, auxin (IAA) inhibited root growth with little effect on hair elongation and significantly stimulated hypocotyl enlargement. Cytokinins (trans-zeatin, kinetin) and ethylene (application of the precursor ACC) likewise exerted an inhibitory effect on root growth but, in contrast to IAA, drastically stimulated root hair elongation. Methylobacteria are phytosymbionts that produce/secrete cytokinins. We conclude that, under real-world conditions (soil), the provision of these phytohormones by methylobacteria (and other epiphytic microbes) regulates root development during seedling establishment.


Subject(s)
Arabidopsis/metabolism , Arabidopsis/microbiology , Methylobacterium/physiology , Plant Roots/metabolism , Plant Roots/microbiology , Arabidopsis/growth & development , Arabidopsis Proteins/metabolism , Cytokinins/metabolism , Indoleacetic Acids/metabolism , Plant Roots/growth & development
11.
Int J Mycobacteriol ; 5(2): 240-3, 2016 06.
Article in English | MEDLINE | ID: mdl-27242240

ABSTRACT

OBJECTIVE/BACKGROUND: A published survey of bacteria in showerhead biofilm samples revealed that Methylobacterium spp. and Mycobacterium spp. seldom coexisted in biofilms. METHODS: To confirm that information, biofilm samples were collected from household plumbing of Mycobacterium avium patients and Methylobacterium spp. and M. avium numbers were measured by direct colony counts. RESULTS: The results demonstrated that if Methylobacterium spp. were present, Mycobacterium spp. were absent, and the opposite. CONCLUSION: The data demonstrate that microbial populations in biofilms can influence the presence or absence of opportunistic premise plumbing pathogens and, thereby, increase the range of strategies to reduce exposure to waterborne pathogens. Finally, by assessing for the visual presence of methylobacteria as pink pigmentation on showers and shower curtains, homeowners and managers of hospitals and other buildings can quickly determine whether a premise plumbing biofilm sample has mycobacteria with a high degree of assurance.


Subject(s)
Methylobacterium/isolation & purification , Mycobacterium/isolation & purification , Sanitary Engineering/instrumentation , Biofilms , Household Articles , Humans , Methylobacterium/growth & development , Methylobacterium/physiology , Mycobacterium/growth & development , Mycobacterium/physiology , Water Microbiology
12.
Arch Microbiol ; 198(1): 17-26, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26475353

ABSTRACT

One-carbon compounds such as methanol, dimethylsulfide (DMS) and dimethylsulfoxide (DMSO) are significant intermediates in biogeochemical cycles. They are suggested to affect atmospheric chemistry and global climate. Methylotrophic microorganisms are considered as a significant sink for these compounds; therefore, we analyzed the diversity of terrestrial bacteria that utilize methanol, DMS and DMSO as carbon and energy source using culture-dependent and culture-independent methods. The effect of habitat type on the methylotrophic community structure was also investigated in rhizosphere and bulk soil. While thirteen strains affiliated to the genera Hyphomicrobium, Methylobacterium, Pseudomonas, Hydrogenophaga, Rhodococcus, Flavobacterium and Variovorax were isolated, denaturing gradient gel electrophoresis revealed the dominance of Thiobacillus, Rhodococcus, Flavobacterium and Bacteroidetes species. Furthermore, methylotrophic communities that degrade methanol or DMS are not shaped by terrestrial habitat type. Rhizosphere and soil samples showed dominance of Methylophilus spp. and Methylovorus spp. for methanol enrichments; Cytophaga spp., Pseudomonas tremae and Thiobacillus thioparus for DMS enrichments.


Subject(s)
Bacteria/classification , Bacteriological Techniques , Biodiversity , Ecosystem , Soil Microbiology , Bacteria/metabolism , Bacteroidetes/physiology , Hyphomicrobium/physiology , Methanol/metabolism , Methylobacterium/physiology
13.
Biocontrol Sci ; 20(2): 91-7, 2015.
Article in English | MEDLINE | ID: mdl-26133506

ABSTRACT

The killing effect of peppermint vapor (PMV) against pink-slime forming microorganisms, Methylobacterium mesophilicum as a bacterium and Rhodotorula mucilaginosa as a yeast, was investigated by the agar vapor assay. In this method, microbial cells were spread over the agar surface exposed to PMV in a petri dish, and then transferred into a recovery liquid. When 60µl of the peppermint liquid was added to a paper disc, a marked killing effect of PMV was observed after 48h against M. mesophilicum and after 168h against R. mucilaginosa. M. mesophilicum and R. mucilaginosa were found to be more resistant to PMV than Escherichia coli and Candida albicans, used as reference microorganisms, respectively. With the addition of 0.03% sodium pyruvate as a hydrogen peroxide scavenger in agar, the killing effect of PMV against E. coli and C. albicans was decreased, whereas it was little changed against M. mesophilicum and R. mucilaginosa. In fact, the properties of the killing effect of hydrogen peroxide solution at 0.2-1.0mM was in accord with those of PMV. M. mesophilicum and R. mucilaginosa were more resistant to the oxidant than E. coli and C. albicans, respectively. Results obtained suggested that reactive oxygen species (ROS) may be involved in the killing action of PMV and therefore pink-slime formers are more resistant to PMV than non-pink-slime formers because of the presence of carotenoids as an antioxidant in cells. We also suggest that the use of PMV appeared to be a potential tool for the control of pink-slime forming microorganisms occurring in wet areas of houses such as the bathroom and washing room.


Subject(s)
Anti-Infective Agents/pharmacology , Biofilms/drug effects , Mentha piperita/chemistry , Methylobacterium/drug effects , Rhodotorula/drug effects , Volatile Organic Compounds/pharmacology , Anti-Infective Agents/isolation & purification , Candida albicans/drug effects , Candida albicans/physiology , Escherichia coli/drug effects , Escherichia coli/physiology , Methylobacterium/physiology , Microbial Viability/drug effects , Rhodotorula/physiology , Volatile Organic Compounds/isolation & purification
14.
PLoS One ; 10(6): e0129509, 2015.
Article in English | MEDLINE | ID: mdl-26053875

ABSTRACT

Methylobacterium species frequently inhabit plant surfaces and are able to utilize the methanol emitted from plants as carbon and energy sources. As some of the Methylobacterium species are known to promote plant growth, significant attention has been paid to the mechanism of growth promotion and the specificity of plant-microbe interactions. By screening our Methylobacterium isolate collection for the high growth promotion effect in vitro, we selected some candidates for field and pot growth tests for rice and barley, respectively. We found that inoculation resulted in better ripening of rice seeds, and increased the size of barley grains but not the total yield. In addition, using whole-cell matrix-assister laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) analysis, we identified and classified Methylobacterium isolates from Methylobacterium-inoculated rice plants. The inoculated species could not be recovered from the rice plants, and in some cases, the Methylobacterium community structure was affected by the inoculation, but not with predomination of the inoculated species. The isolates from non-inoculated barley of various cultivars grown in the same field fell into just two species. These results suggest that there is a strong selection pressure at the species level of Methylobacterium residing on a given plant species, and that selection of appropriate species that can persist on the plant is important to achieve growth promotion.


Subject(s)
Hordeum/growth & development , Hordeum/microbiology , Methylobacterium/physiology , Oryza/growth & development , Oryza/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Likelihood Functions , Methylobacterium/genetics , Methylobacterium/isolation & purification , Molecular Sequence Data , Phylogeny , Plant Leaves/microbiology , RNA, Ribosomal, 16S/genetics , Seeds/microbiology , Species Specificity
15.
Biomed Res Int ; 2015: 909016, 2015.
Article in English | MEDLINE | ID: mdl-25861650

ABSTRACT

The genus Methylobacterium is composed of pink-pigmented facultative methylotrophic (PPFM) bacteria, which are able to synthesize carotenoids and grow on reduced organic compounds containing one carbon (C1), such as methanol and methylamine. Due to their high phenotypic plasticity, these bacteria are able to colonize different habitats, such as soil, water, and sediment, and different host plants as both endophytes and epiphytes. In plant colonization, the frequency and distribution may be influenced by plant genotype or by interactions with other associated microorganisms, which may result in increasing plant fitness. In this review, different aspects of interactions with the host plant are discussed, including their capacity to fix nitrogen, nodule the host plant, produce cytokinins, auxin and enzymes involved in the induction of systemic resistance, such as pectinase and cellulase, and therefore plant growth promotion. In addition, bacteria belonging to this group can be used to reduce environmental contamination because they are able to degrade toxic compounds, tolerate high heavy metal concentrations, and increase plant tolerance to these compounds. Moreover, genome sequencing and omics approaches have revealed genes related to plant-bacteria interactions that may be important for developing strains able to promote plant growth and protection against phytopathogens.


Subject(s)
Host-Pathogen Interactions/physiology , Methylobacterium/metabolism , Methylobacterium/physiology , Plants/microbiology , Biotechnology/methods , Carbon/metabolism , Ecosystem , Humans
16.
Microbes Environ ; 29(4): 388-92, 2014.
Article in English | MEDLINE | ID: mdl-25381715

ABSTRACT

Concern regarding household biofilms has grown due to their widespread existence and potential to threaten human health by serving as pathogen reservoirs. Previous studies identified Methylobacterium as one of the dominant genera found in household biofilms. In the present study, we examined the mechanisms underlying biofilm formation by using the bacterial consortium found in household pink slime. A clone library analysis revealed that Methylobacterium was the predominant genus in household pink slime. In addition, 16 out of 21 pink-pigmented bacterial isolates were assigned to the genus Methylobacterium. Although all of the Methylobacterium isolates formed low-level biofilms, the amount of the biofilms formed by Methylobacterium sp. P-1M and P-18S was significantly increased by co-culturing with other Methylobacterium strains that belonged to a specific phylogenetic group. The single-species biofilm was easily washed from the glass surface, whereas the dual-species biofilm strongly adhered after washing. A confocal laser scanning microscopy analysis showed that the dual-species biofilms were significantly thicker and tighter than the single-species biofilms.


Subject(s)
Biofilms/growth & development , Environmental Microbiology , Methylobacterium/physiology , Microbial Interactions , Pigments, Biological , Cluster Analysis , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Family Characteristics , Humans , Methylobacterium/classification , Methylobacterium/growth & development , Methylobacterium/isolation & purification , Microscopy, Confocal , Molecular Sequence Data , Phylogeny , Pigments, Biological/analysis , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA
17.
Tree Physiol ; 34(9): 993-1005, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25149086

ABSTRACT

Tissues of Scots pine (Pinus sylvestris L.) contain several endophytic microorganisms of which Methylobacterium extorquens DSM13060 is a dominant species throughout the year. Similar to other endophytic bacteria, M. extorquens is able to colonize host plant tissues without causing any symptoms of disease. In addition to endophytic bacteria, plants associate simultaneously with a diverse set of microorganisms. Furthermore, plant-colonizing microorganisms interact with each other in a species- or strain-specific manner. Several studies on beneficial microorganisms interacting with plants have been carried out, but few deal with interactions between different symbiotic organisms and specifically, how these interactions affect the growth and development of the host plant. Our aim was to study how the pine endophyte M. extorquens DSM13060 affects pine seedlings and how the co-inoculation with ectomycorrhizal (ECM) fungi [Suillus variegatus (SV) or Pisolithus tinctorius (PT)] alters the response of Scots pine. We determined the growth, polyamine and nutrient contents of inoculated and non-inoculated Scots pine seedlings in vitro. Our results show that M. extorquens is able to improve the growth of seedlings at the same level as the ECM fungi SV and PT do. The effect of co-inoculation using different symbiotic organisms was seen in terms of changes in growth and nutrient uptake. Inoculation using M. extorquens together with ECM fungi improved the growth of the host plant even more than single ECM inoculation. Symbiotic organisms also had a strong effect on the potassium content of the seedling. The results indicate that interaction between endophyte and ECM fungus is species dependent, leading to increased or decreased nutrient content and growth of pine seedlings.


Subject(s)
Basidiomycota/physiology , Methylobacterium/physiology , Pinus sylvestris/growth & development , Pinus sylvestris/microbiology , Polyamines/metabolism , Endophytes/physiology , Mycorrhizae/physiology , Pinus sylvestris/metabolism , Plant Roots/growth & development , Plant Roots/metabolism , Plant Roots/microbiology , Seedlings/growth & development , Seedlings/metabolism , Seedlings/microbiology
18.
Naturwissenschaften ; 101(4): 339-46, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24566997

ABSTRACT

On the leaf surfaces of numerous plant species, inclusive of sunflower (Helianthus annuus L.), pink-pigmented, methanol-consuming, phytohormone-secreting prokaryotes of the genus Methylobacterium have been detected. However, neither the roles, nor the exact mode of colonization of these epiphytic microbes have been explored in detail. Using germ-free sunflower seeds, we document that, during the first days of seedling development, methylobacteria exert no promotive effect on organ growth. Since the microbes are evenly distributed over the outer surface of the above-ground phytosphere, we analyzed the behavior of populations taken from two bacterial strains that were cultivated as solid, biofilm-like clones on agar plates in different aqueous environments (Methylobacterium mesophilicum and M. marchantiae, respectively). After transfer into liquid medium, the rod-shaped, immobile methylobacteria assembled a flagellum and developed into planktonic microbes that were motile. During the linear phase of microbial growth in liquid cultures, the percentage of swimming, flagellated bacteria reached a maximum, and thereafter declined. In stationary populations, living, immotile bacteria, and isolated flagella were observed. Hence, methylobacteria that live in a biofilm, transferred into aqueous environments, assemble a flagellum that is lost when cell density has reached a maximum. This swimming motility, which appeared during ontogenetic development within growing microbial populations, may be a means to colonize the moist outer surfaces of leaves.


Subject(s)
Flagella/physiology , Helianthus/microbiology , Methylobacterium/physiology , Biofilms , Methylobacterium/cytology , Plant Leaves/microbiology , Population Density
19.
J Appl Microbiol ; 116(2): 408-23, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24188110

ABSTRACT

AIM: To develop co-aggregated bacterial inoculant comprising of Methylobacterium oryzae CBMB20/Methylobacterium suomiense CBMB120 strains with Azospirillum brasilense (CW903) strain and testing their efficiency as inoculants for plant growth promotion (PGP). METHODS AND RESULTS: Biofilm formation and co-aggregation efficiency was studied between A. brasilense CW903 and methylobacterial strains M. oryzae CBMB20 and M. suomiense CBMB120. Survival and release of these co-aggregated bacterial strains entrapped in alginate beads were assessed. PGP attributes of the co-aggregated bacterial inoculant were tested in tomato plants under water-stressed conditions. Results suggest that the biofilm formation efficiency of the CBMB20 and CBMB120 strains increased by 15 and 34%, respectively, when co-cultivated with CW903. Co-aggregation with CW903 enhanced the survivability of CBMB20 strain in alginate beads. Water stress index score showed least stress index in plants inoculated with CW903 and CBMB20 strains maintained as a co-aggregated inoculant. CONCLUSIONS: This study reports the development of co-aggregated cell inoculants containing M. oryzae CBMB20 and A. brasilense CW903 strains conferred better shelf life and stress abatement in inoculated tomato plants. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings could be extended to other PGP bacterial species to develop multigeneric bioinoculants with multiple benefits for various crops.


Subject(s)
Alginates/chemistry , Azospirillum brasilense/physiology , Biofilms/growth & development , Methylobacterium/physiology , Solanum lycopersicum/growth & development , Azospirillum brasilense/enzymology , Azospirillum brasilense/ultrastructure , Dehydration/prevention & control , Droughts , Ethylenes/metabolism , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Hydrolysis , Lipid Peroxidation , Solanum lycopersicum/microbiology , Malondialdehyde/metabolism , Methylobacterium/enzymology , Methylobacterium/ultrastructure , Microscopy, Electron, Scanning , Microspheres , Peroxidase/metabolism , Soil/chemistry
20.
Braz. j. microbiol ; 44(4): 1331-1339, Oct.-Dec. 2013. graf, tab
Article in English | LILACS | ID: lil-705276

ABSTRACT

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.


Subject(s)
Acyl-Butyrolactones/metabolism , Gene Expression Regulation, Bacterial/drug effects , Host-Parasite Interactions , Methylobacterium/physiology , Plant Extracts/metabolism , Plants/microbiology , Methylobacterium/growth & development
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