Structural characterization of lipid A obtained from Pantoea agglomerans lipopolysaccharide
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Lipopolysaccharide isolated from Pantoea agglomerans showed higher priming and triggering activities for macrophages in terms of tumor necrosis factor production than other lipopolysaccharides. To identify the difference in biological activities of lipopolysaccharide of Pantoea agglomerans from other lipopolysaccharides on the basis of structure, we determined the structure of the lipid A part, which is the biological center of lipopolysaccharide, by quantitative analysis, nuclear magnetic resonance spectroscopy and mass spectrometry. Lipopolysaccharide of Pantoea agglomerans is constructed with at least two kinds of lipid A of different levels of acylation. One is of the same type as that of Escherichia coli with hexa-acyl lipid A and the other is the Salmonella minnesota type with hepta-acyl lipid A.Keywords:
Pantoea agglomerans
Lipid A
Pantoea
The genus Pantoea forms a complex of more than 25 species, among which several cause diseases of various crop plants, including rice. Notably, strains of Pantoea ananatis and P. stewartii have been repeatedly reported to cause bacterial leaf blight of rice, whereas other authors have observed that P. agglomerans can also cause bacterial leaf blight of rice. The contribution of these and perhaps other species of Pantoea to plant diseases and yield losses of crop plants is currently not well documented, partly due to the lack of efficient diagnostic tools. Using 32 whole-genome sequences of the three major plant-pathogenic Pantoea spp., a set of PCR primers that detect each of the three species P. agglomerans, P. ananatis, and P. stewartii was designed. A multiplex PCR scheme which can distinguish these three species and also detects members of other Pantoea spp. was further developed. Upon validation on a set of reference strains, 607 suspected Pantoea strains that were isolated from rice leaves or seed originating from 11 African countries were screened. In total, 41 P. agglomerans strains from 8 countries, 79 P. ananatis strains from 9 countries, 269 P. stewartii strains from 9 countries, and 218 unresolved Pantoea strains from 10 countries were identified. The PCR protocol allowed detection of Pantoea bacteria grown in vitro, in planta, and in rice seed. The detection threshold was estimated as total genomic DNA at 0.5 ng/µl and heated cells at 1 × 104 CFU/ml. This new molecular diagnostic tool will help to accurately diagnose major plant-pathogenic species of Pantoea. Due to its robustness, specificity, sensitivity, and cost efficiency, it will be very useful for plant protection services and for the epidemiological surveillance of these important crop-threatening bacteria.
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Abstract The genus Pantoea incorporates many economically and clinically important species. The plant-associated species, Pantoea agglomerans and Pantoea vagans, are closely related and are often isolated from similar environments. Plasmids conferring certain metabolic capabilities are also shared amongst these two species. The genomes of two isolates obtained from fungus-growing termites in South Africa were sequenced, assembled and annotated. A high number of orthologous genes are conserved within and between these species. The difference in genome size between P. agglomerans MP2 (4,733,829 bp) and P. vagans MP7 (4,598,703 bp) can largely be attributed to the differences in plasmid content. The genome sequences of these isolates may shed light on the common traits that enable P. agglomerans and P. vagans to co-occur in plant- and insect-associated niches.
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The Enterobacterial genus Pantoea contains both free-living and host-associating species, with considerable debate as to whether documented reports of human infections by members of this species group are accurate. MALDI-TOF-based identification methods are commonly used in clinical laboratories as a rapid means of identification, but its reliability for identification of Pantoea species is unclear. In this study, we carried out cpn60-based molecular typing of 54 clinical isolates that had been identified as Pantoea using MALDI-TOF and other clinical typing methods. We found that 24% had been misidentified, and were actually strains of Citrobacter, Enterobacter, Kosakonia, Klebsiella, Pseudocitrobacter, members of the newly described Erwinia gerundensis, and even several unclassified members of the Enterobacteriaceae. The 40 clinical strains that were confirmed to be Pantoea were identified as Pantoea agglomerans, Pantoea allii, Pantoea dispersa, Pantoea eucalypti, and Pantoea septica as well as the proposed species group, Pantoea latae. Some species groups considered largely environmental or plant-associated, such as P. allii and P. eucalypti were also among clinical specimens. Our results indicate that MALDI-TOF-based identification methods may misidentify strains of the Enterobacteriaceae as Pantoea.
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Abstract Background The genus Pantoea forms a complex of more than 25 species, among which several cause diseases of several crop plants, including rice. Notably, strains of Pantoea ananatis and Pantoea stewartii have been found to cause bacterial leaf blight of rice in Togo and Benin, while other authors have observed that Pantoea agglomerans can also cause bacterial leaf blight of rice. The contribution of these and perhaps other species of Pantoea to plant diseases and yield losses of crop plants is currently not well documented, partly due to the lack of efficient diagnostic tools. Result Using 34 whole genome sequences of the three-major plant-pathogenic Pantoea species, a set of PCR primers that specifically detect each of the three species, P. agglomerans , P. ananatis , and P. stewartii , was designed. A multiplex PCR protocol which can distinguish these three species and also detects members of other Pantoea species was further developed. Upon validation on a set of reference strains, 609 suspected Pantoea strains that were isolated from rice leaves or seeds originating from 11 African countries were screened. In total, 41 P. agglomerans strains from eight countries, 79 P. ananatis strains from nine countries, 269 P. stewartii strains from nine countries and 220 unsolved Pantoea strains from ten countries were identified. The PCR protocol allowed detecting Pantoea bacteria grown in vitro, in planta and in rice seeds. The detection threshold was estimated at 5 ng/mL of total genomic DNA and 1 × 10 5 CFU/mL of heated cells. Conclusion This new molecular diagnostic tool will help accurately diagnose major plant-pathogenic species of Pantoea . Due to its robustness, specificity, sensitivity, and cost efficiency it will be very useful for plant protection services and for the epidemiological surveillance of these important crop-threatening bacteria.
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Abstract Pantoea spp. is one of the opportunistic pathogens, which causes the spread of hospital infection due to its transformation from bacteria to plant organisms to human pathogenic bacteria. A total of 623 samples were collected and distributed as 483 samples of different clinical cases (77.52%) and 140 environmental samples (22.47%) from Diwaniyah city hospitals. The results showed that 24 isolates of pantoea spp.s had been confirmed using API-20E Polymerase chain reaction technology. The current study reported the presence of two types of pathogens for the Pantoea spp.: the first is Pantoea agglomerans and Pantoea calida in hospitals in the city of Diwaniyah. Pantoea agglomerans was occurring in the studied isolates at a higher rate (2.27%) than Pantoea calida. All isolates of Pantoea spp were tested for the prevalence of virulence genes (avrxacE2, avrxacEl, hrpG, acrAB) using the polymerase chain reaction (PCR) technique. The highest incidence of hrpG was recorded as 79.16%, followed by avraxacE2 gene were 13 (54.16%). The current study did not record any presence of the virulence genes (avrxacE1 acrAB) among the isolates of Pantoea spp.
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Lipopolysaccharide isolated from Pantoea agglomerans showed higher priming and triggering activities for macrophages in terms of tumor necrosis factor production than other lipopolysaccharides. To identify the difference in biological activities of lipopolysaccharide of Pantoea agglomerans from other lipopolysaccharides on the basis of structure, we determined the structure of the lipid A part, which is the biological center of lipopolysaccharides, by quantitative analysis, nuclear magnetic resonance spectroscopy and mass spectrometry. Lipopolysaccharide of Pantoea agglomerans is constructed with at least two kinds of lipid A of different levels of acylation. One is of the same type as that of Escherichia coli with hexa-acyl lipid A and the other is the Salmonella minnesota type with hepta-acyl lipid A.
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Bacteria isolated from eucalyptus leaves and shoots showing symptoms of blight and die-back collected in Uganda, Uruguay and Argentina and from maize displaying brown stalk rot symptoms in South Africa were tentatively placed in the genus Pantoea on the basis of phenotypic and biochemical tests. These isolates, together with two strains (LMG 2558 and LMG 2560) previously assigned to Pantoea agglomerans based on protein electrophoregrams but later excluded from this species, were further investigated using molecular techniques. 16S rRNA gene sequencing and multilocus sequence analyses (MLSA) revealed that the strains were phylogenetically closely related to Pantoea agglomerans, Pantoea stewartii and Pantoea ananatis. MLSA and amplified fragment length polymorphism analysis placed the strains into four separate clusters, not containing any of the type strains of species of the genus Pantoea. DNA–DNA hybridization confirmed the classification of the isolates into four novel species, for which the names Pantoea vagans sp. nov. (type strain R-21566T =LMG 24199T =BCC 105T =BD 765T), Pantoea eucalypti sp. nov. (type strain R-25678T =LMG 24197T =BCC 076T =BD 769T), Pantoea deleyi sp. nov. (type strain R-31523T =LMG 24200T =BCC 109T =BD 767T) and Pantoea anthophila sp. nov. (type strain LMG 2558T =BD 871T =NCPPB 1682T) are proposed.
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The ubiquitous Gram-negative bacterium Pantoea agglomerans (synonyms: Enterobacter agglomerans, Erwinia herbicola) is known both as an epiphytic microbe developing on the surface of plants and as an endophytic organism living inside the plants. The bacterium occurs also abundantly in plant and animal products, in the body of arthropods and other animals, in water, soil, dust and air, and occasionally in humans. From the human viewpoint, the role of this organism is ambiguous, both deleterious and beneficial: on one side it causes disorders in people exposed to inhalation of organic dusts and diseases of crops, and on the other side it produces substances effective in the treatment of cancer and other diseases of humans and animals, suppresses the development of various plant pathogens, promotes plant growth, and appears as a potentially efficient biofertilizer and bioremediator. P. agglomerans was identified as a predominant bacterium on cotton plant grown all over the world, usually as an epiphyte, rarely as pathogen. It is particularly numerous on cotton bract after senescence. During processing of cotton in mills, bacteria and their products are released with cotton dust into air and are inhaled by workers, causing respiratory and general disorders, usually defined as byssinosis. The most adverse substance is endotoxin, a heteropolymer macromolecule present in the outermost part of the cell wall, consisting of lipopolysaccharide (LPS) as a major constituent, phospholipids and protein. The numerous experiments carried out in last quarter of XXth century on laboratory animals and human volunteers supported a convincing evidence that the inhaled endotoxin produced by P. agglomerans causes numerous pathologic effects similar to those elicited by cotton dust, such as influx of free lung cells into airways and activation of alveolar macrophages which secrete mediators (prostaglandins, platelet-activating factor, interleukin-1, tumor necrosis factor) that cause accumulation of platelets in pulmonary capillaries initiating an acute and chronic inflammation resulting in endothelial cell damage and extravasation of cells and fluids into the lung interstitium. These changes cause bronchoconstriction, the decrement of lung function expressed as reduction of forced expiratory volume in one second (FEV1) and/or diffusion capacity, increase in the airway hyperreactivity and subjective symptoms such as fever, airway irritation and chest tightness. The conclusions from these experiments, performed mostly 2-3 decades ago, did not loose their actuality until recently as so far no other cotton dust component was identified as a more important work-related hazard than bacterial endotoxin. Though also other microbial and plant constituents are considered as potential causative agents of byssinosis, the endotoxin produced by Pantoea agglomerans and other Gram-negative bacteria present in cotton dust is still regarded as a major cause of this mysterious disease.
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Abstract The effect of N-acyl-homoserine lactone (AHL)-based quorum sensing (QS) on biogenic amine (BA) formation by Pantoea agglomerans was studied. Agrobacterium tumefaciens A136 and KYC55-based bioassays confirmed AHLs production by Pantoea agglomerans . The production ability of AHLs was quantified on the basis of β-galactosidase activity. The influence of temperature (10°C and 20°C) and pH (5.5 and 6.5) on β-galactosidase activity and BAs production by Pantoea agglomerans was determined. Acidification of the environment adversely affected the growth and β-galactosidase activity of Pantoea agglomerans , and AHLs production and BAs accumulation by Pantoea agglomerans was inhibited at low temperature. A significant correlation between β-galactosidase activity and BAs (putrescine, histamine, putrescine and tryptamine) was identified ( P < 0.01). Based on the results of this study, the AHL-based QS system influences the concentrations and types of BAs produced by Pantoea agglomerans .
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Lipopolysaccharide isolated from Pantoea agglomerans showed higher priming and triggering activities for macrophages in terms of tumor necrosis factor production than other lipopolysaccharides. To identify the difference in biological activities of lipopolysaccharide of Pantoea agglomerans from other lipopolysaccharides on the basis of structure, we determined the structure of the lipid A part, which is the biological center of lipopolysaccharide, by quantitative analysis, nuclear magnetic resonance spectroscopy and mass spectrometry. Lipopolysaccharide of Pantoea agglomerans is constructed with at least two kinds of lipid A of different levels of acylation. One is of the same type as that of Escherichia coli with hexa-acyl lipid A and the other is the Salmonella minnesota type with hepta-acyl lipid A.
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