Exploring the pathogenic function of Pantoea ananatis endogenous plasmid by an efficient and simple plasmid elimination strategy.

The bacterium Pantoea ananatis is associated with devastating plant diseases that cause serious economic losses. Strain DZ-12 was previously isolated from maize brown rot leaves in Hebei Province, China and its genome sequencing revealed that it belongs to P. ananatis. It contains a large, endogenous plasmid, pDZ-12. Different studies have shown that virulence determinants are frequently carried on plasmids. To determine whether pDZ-12 from P. ananatis has any effect on pathogenicity, the plasmid was eliminated by substituting its native replication genes with temperature-sensitive replication genes. The resulting temperature-sensitive plasmid could be cured by growing cells at high temperature (37℃). Loss of pDZ-12 from P. ananatis DZ-12 led to a decreased disease severity in maize plants suggesting that the endogenous plasmid is important for pathogenesis. Loss of pDZ-12 also affected the ability of the bacterium to form biofilms. The study provides the first evidence that the endogenous plasmid of P. ananatis DZ-12 is important for pathogenesis in maize plants and carries genes involved in biofilm formation. This study also presents the first report on curing a plasmid from P. ananatis.

Comprehensive genomic analysis reveals virulence factors and antibiotic resistance genes in Pantoea agglomerans KM1, a potential opportunistic pathogen.

Pantoea agglomerans is a Gram-negative facultative anaerobic bacillus causing a wide range of opportunistic infections in humans including septicemia, pneumonia, septic arthritis, wound infections and meningitis. To date, the determinants of virulence, antibiotic resistance, metabolic features conferring survival and host-associated pathogenic potential of this bacterium remain largely underexplored. In this study, we sequenced and assembled the whole-genome of P. agglomerans KM1 isolated from kimchi in South Korea. The genome contained one circular chromosome of 4,039,945 bp, 3 mega plasmids, and 2 prophages. The phage-derived genes encoded integrase, lysozyme and terminase. Six CRISPR loci were identified within the bacterial chromosome. Further in-depth analysis showed that the genome contained 13 antibiotic resistance genes conferring resistance to clinically important antibiotics such as penicillin G, bacitracin, rifampicin, vancomycin, and fosfomycin. Genes involved in adaptations to environmental stress were also identified which included factors providing resistance to osmotic lysis, oxidative stress, as well as heat and cold shock. The genomic analysis of virulence factors led to identification of a type VI secretion system, hemolysin, filamentous hemagglutinin, and genes involved in iron uptake and sequestration. Finally, the data provided here show that, the KM1 isolate exerted strong immunostimulatory properties on RAW 264.7 macrophages in vitro. Stimulated cells produced Nitric Oxide (NO) and pro-inflammatory cytokines TNF-α, IL-6 and the anti-inflammatory cytokine IL-10. The upstream signaling for production of TNF-α, IL-6, IL-10, and NO depended on TLR4 and TLR1/2. While production of TNF-α, IL-6 and NO involved solely activation of the NF-κB, IL-10 secretion was largely dependent on NF-κB and to a lesser extent on MAPK Kinases. Taken together, the analysis of the whole-genome and immunostimulatory properties provided in-depth characterization of the P. agglomerans KM1 isolate shedding a new light on determinants of virulence that drive its interactions with the environment, other microorganisms and eukaryotic hosts.

Retention of Pantoea agglomerans Sc1R across stadia of the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae).

Southern green stink bug [Nezara viridula (L.)] adults and other pentatomid pests can transmit pathogens (e.g., the bacterium Pantoea agglomerans) that cause disease in cotton (Gossypium hirsutum L.) and other high-value cash crops worldwide. First instars of N. viridula were recently shown to ingest P. agglomerans strain Sc1R with rifampicin-resistance, and to retain the pathogen to the 2nd instar. The objective of this study was to determine the acquisition of P. agglomerans Sc1R by early instars of N. viridula and determine persistence of P. agglomerans Sc1R across subsequent stadia. In three trials, early instars (1st and 2nd) were exposed to P. agglomerans Sc1R and subsequently maintained to adulthood; cohorts were sampled at 3rd and 5th instars, as well as adults. In every trial, P. agglomerans Sc1R was detected in all stadia, including adults, but significantly higher frequencies of infection than expected were observed at the initial stage of infection (either 1st or 2nd instar). Higher densities of P. agglomerans Sc1R were detected in 1st and 2nd instars, and lower densities were observed in subsequent stadia. Densities of innate microbiota were generally lower when the initial stage of exposure was at 1st instar than when the initial stage of exposure was at the 2nd instar. Overall, half of the adults possessed P. agglomerans Sc1R. These findings demonstrated that N. viridula nymphs can acquire P. agglomerans Sc1R and retain the pathogen to adulthood. Potential avenues of research to further elucidate the implications of nymphs harboring pathogens to adulthood are discussed.

Thiosulfinate Tolerance Is a Virulence Strategy of an Atypical Bacterial Pathogen of Onion.

Unlike most characterized bacterial plant pathogens, the broad-host-range plant pathogen Pantoea ananatis lacks both the virulence-associated type III and type II secretion systems. In the absence of these typical pathogenicity factors, P. ananatis induces necrotic symptoms and extensive cell death in onion tissue dependent on the HiVir proposed secondary metabolite synthesis gene cluster. Onion (Allium. cepa L), garlic (A. sativum L.), and other members of the Allium genus produce volatile antimicrobial thiosulfinates upon cellular damage. However, the roles of endogenous thiosulfinate production in host-bacterial pathogen interactions have not been described. We found a strong correlation between the genetic requirements for P. ananatis to colonize necrotized onion tissue and its capacity for tolerance to the thiosulfinate "allicin" based on the presence of an eleven-gene, plasmid-borne, virulence cluster of sulfur redox genes. We have designated them "alt" genes for allicin tolerance. We show that allicin and onion thiosulfinates restrict bacterial growth with similar kinetics. The alt gene cluster is sufficient to confer allicin tolerance and protects the glutathione pool during allicin treatment. Independent alt genes make partial phenotypic contributions indicating that they function as a collective cohort to manage thiol stress. Our work implicates endogenous onion thiosulfinates produced during cellular damage as major mediators of interactions with bacteria. The P. ananatis-onion pathosystem can be modeled as a chemical arms race of pathogen attack, host chemical counterattack, and pathogen defense.

Genetic diversity of Pantoea stewartii subspecies stewartii causing jackfruit-bronzing disease in Malaysia.

Jackfruit-bronzing is caused by bacteria Pantoea stewartii subspecies stewartii (P. stewartii subsp. stewartii), showing symptoms of yellowish-orange to reddish discolouration and rusty specks on pulps and rags of jackfruit. Twenty-eight pure bacterial strains were collected from four different jackfruit outbreak collection areas in Peninsular Malaysia (Jenderam, Maran, Muadzam Shah and Ipoh). Positive P. stewartii subsp. stewartii verification obtained in the study was based on the phenotypic, hypersensitivity, pathogenicity and molecular tests. Multilocus sequence analysis (MLSA) was performed using four housekeeping genes (gyrB, rpoB, atpD and infB) on all 28 bacterial strains. Single gyrB, rpoB, atpD and infB phylogenetic trees analyses revealed the bootstrap value of 99-100% between our bacterial strains with P. stewartii subsp. stewartii reference strains and P. stewartii subsp. indologenes reference strains. On the other hand, phylogenetic tree of the concatenated sequences of the four housekeeping genes revealed that our 28 bacterial strains were more closely related to P. stewartii subsp. stewartii (99% similarities) compared to its close relative P. stewartii subsp. indologenes, although sequence similarity between these two subspecies were up to 100%. All the strains collected from the four collection areas clustered together, pointing to no variation among the bacterial strains. This study improves our understanding and provided new insight on the genetic diversity of P. stewartii subsp. stewartii associated with jackfruit-bronzing in Malaysia.

Infección del tracto urinario por Pantoea agglomerans: ¿un patógeno de pacientes inmunodeprimidos? / Urinary tract infection by Pantoea agglomerans: a pathogen of immunosuppressed patients?

La infección por Pantoea agglomerans es muy rara de forma global y particularmente en Pediatría. La mayoría de las infecciones que causa son nosocomiales en pacientes inmunodeprimidos. Se han descrito infecciones articulares tras punción con espinas vegetales dando lugar a artritis, sinovitis u osteítis; meningitis o septicemia neonatal tras la contaminación de la nutrición parenteral o transfusiones de hematíes; infecciones de orina o respiratorias y peritonitis. En inmunocompetentes las infecciones son excepcionales, por lo que se aconseja realizar estudio de inmunidad, como en nuestro caso. El germen es muy resistente a antibióticos betalactámicos y para erradicarlo suele ser necesario tratar según antibiograma

Pantoea agglomerans infections are very rare globally and particularly in Pediatrics. Most of them are nosocomial and affect immnunosuppresed patients. Articular infections have been described after puncture with plant thorns giving rise to arthritis, synovitis or osteitis; they also can cause meningitis or neonatal septicemia after contamination of parenteral nutrition or red blood cell transfusions; urinary or respiratory infections, and peritonitis. In immunocompetent patients the infections are exceptional, so it is advisable to perform an immunity study, as in our case. The germ is very resistant to beta-lactam antibiotics and to eradicate it, it is usually necessary to treat according to antibiogram

Pathogenicity and a TaqMan Real-Time PCR for Specific Detection of Pantoea allii, a Bacterial Pathogen of Onions.

Bacterial diseases of onion are reported to cause significant economic losses. Pantoea allii Brady, one of the pathogens causing the center rot on onions, has not yet been reported in Canada. We report the pathogenicity of P. allii on commercially available Canadian green onions (scallions). All P. allii-inoculated plants, irrespective of the inoculum concentration, exhibited typical leaf chlorotic discoloration on green onion leaves, which can reduce their marketability. Reisolation of P. allii from infected scallion tissues and reidentification by sequencing and phylogenetic analyses of the leuS gene suggest that the pathogen can survive in infected tissues 21 days after inoculation. This is the first report of P. allii as a potential pathogen of green onions. This study also reports the development and validation of a TaqMan real-time PCR assay targeting the leuS gene for reliable detection of P. allii in pure cultures and in planta. A 642-bp leuS gene fragment was targeted because it showed high nucleotide diversity and positively correlated with genome-based average nucleotide identity with respect to percent similarity index and identity of Pantoea species. The assay specificity was validated using 61 bacterial and fungal strains. Under optimal conditions, the selected primers and FAM-labeled TaqMan probe were specific for the detection of nine reference P. allii strains by real-time PCR. The 52 strains of other Pantoea spp. (n = 25), non-Pantoea spp. (n = 20), and fungi/oomycetes (n = 7) tested negative (no detectable fluorescence). Onion tissues spiked with P. allii, naturally infested onion bulbs, greenhouse infected green onion leaf samples, as well as an interlaboratory blind test were used to validate the assay specificity. The sensitivities of a 1-pg DNA concentration and 30 CFU are comparable to previously reported real-time PCR assays of other bacterial pathogens. The TaqMan real-time PCR assay developed in this study will facilitate reliable detection of P. allii and could be a useful tool for screening onion imports or exports for the presence of this pathogen.

Atividade de nucleação de gelo e densidade populacional do patógeno podem ser fatores que contribuem para o progresso da doença mancha branca do milho / Ice nucleation activity and density of the pathogen population may be factors that contribute to the progression of maize white spot disease / La actividad de nucleación de hielo y la densidad de población del patógeno pueden ser factores que contribuyen al progreso de la enfermedad mancha blanca del maíz

A Mancha Branca do Milho (MBM) é uma doença foliar que tem causado perdas apreciáveis, tanto qualitativas como quantitativas para a produção de milho. Seu agente etiológico, a Pantoea ananatis, é uma bactéria epifítica, Gram-negativa formadora de colônia amarela, capaz de formar gelo, mesmo em ambiente tropical (ice nucleation activity - INA). Este estudo teve por objetivo caracterizar isolados de P. ananatis quanto à atividade INA e avaliar o efeito da densidade bacteriana na expressão do fenótipo INA e no processo de desenvolvimento da doença. O agente patogênico foi isolado de lesões iniciais da doença, as anasarcas, e avaliado quanto à expressão do fenótipo INA em diferentes concentrações bacterianas. O mesmo foi feito com isolados epifíticos obtidos da superfície foliar de plantas de milho sadias. Dos 24 isolados bacterianos estudados, apenas 13 apresentaram o fenótipo INA+. A expressão deste fenótipo foi dependente da densidade celular. Isolados INA+ e INA­ foram inoculados em folhas destacadas e em plantas da cultivar HS200 a campo, em diferentes concentrações do inóculo. Nenhum isolado INA­ reproduziu sintomas em laboratório. Dos cinco isolados INA+ somente um deles reproduziu sintomas em laboratório. Em campo o isolado INA+ foi capaz de promover lesões em todas as concentrações avaliadas. Conclui-se que a atividade de nucleação de gelo pela bactéria P. ananatis é dependente da linhagem e da densidade bacteriana e este fenômeno pode estar envolvido no desenvolvimento dos sintomas da MBM.(AU)

Maize White Spot Disease is a leaf disease that has caused considerable losses, both qualitative and quantitative for corn production. Its etiologic agent, Pantoea ananatis, is an epiphytic, Gram-negative, yellow colony-forming bacterium, capable of forming ice, even in tropical environments at temperatures where this normally does not occur (Ice Nucleation Activity - INA). This study aimed at characterizing P. ananatis isolates in terms of INA activity and evaluating the effect of bacterial density on the expression of the INA phenotype and on the disease development process. The pathogen was isolated from the initial lesions of the disease, the anasarcas, and were evaluated for the expression of the INA phenotype in different bacterial concentrations. The same procedure was performed on epiphytic isolates obtained from the leaf surface of healthy maize plants. From the 24 bacterial isolates studied, only 13 presented the INA+ phenotype. The expression of this phenotype is dependent on cell density. INA+ and INA­ isolates were inoculated on detached leaves and on plants of cultivar HS200 in the field, in different concentrations of the inoculum. No INA­ isolates reproduced symptoms in the laboratory. From the five INA+ isolates, only one of them reproduced symptoms in the laboratory. In the field, the INA+ isolate was able to promote lesions in all concentrations evaluated. It can be concluded that the ice nucleation activity by P. ananatis is dependent on the strain and bacterial density and this phenomenon may be involved in the development of Maize White Spot Disease symptoms.(AU)

La mancha blanca del maíz es una enfermedad de la hoja que ha causado pérdidas considerables, tanto cualitativas como cuantitativas para la producción de maíz. Su agente etiológico, Pantoea ananatis, es una bacteria epífita, Gram-negativa, formadora de colonias amarillas, capaz de causar hielo, incluso en ambientes tropicales a temperaturas donde esto normalmente no ocurre (actividad de nucleación de hielo - INA). Este estudio tuvo como objetivo caracterizar los aislados de P. ananatis en términos de actividad de INA y evaluar el efecto de la densidad bacteriana en la expresión del fenotipo de INA y en el proceso de desarrollo de la enfermedad. El patógeno se aisló de las lesiones iniciales de la enfermedad, las anasarcas, y se evaluó la expresión del fenotipo INA en diferentes concentraciones bacterianas. Lo mismo se hizo con aislamientos epifitos obtenidos de la superficie de la hoja de plantas de maíz sanas. De los 24 aislados bacterianos estudiados, solo 13 presentaron el fenotipo INA+. La expresión de este fenotipo depende de la densidad celular. Se inocularon aislamientos INA+ e INA- en hojas desprendidas y en plantas del cultivar HS200 en el campo, en diferentes concentraciones del inóculo. Ninguno aislado INA- reprodujo síntomas en el laboratorio. De los cinco aislamientos de INA+, solo uno de ellos reprodujo síntomas en el laboratorio. En el campo, el aislado INA+ pudo promover lesiones en todas las concentraciones evaluadas. Se concluye que la nucleación de hielo por P. ananatis depende de la cepa y la densidad bacteriana, y este fenómeno puede estar involucrado en el desarrollo de los síntomas de la enfermedad de la mancha blanca del maíz.(AU)

Pantoea agglomerans as a New Etiological Agent of a Bacterial Necrotic Disease of Mango Trees.

Bacterial apical necrosis of mango trees, a disease elicited by Pseudomonas syringae pv. syringae, is a primary limiting factor of mango crop production in the Mediterranean region. In this study, a collection of bacterial isolates associated with necrotic symptoms in mango trees similar to those produced by bacterial apical necrosis disease were isolated over five consecutive years in orchards from the Canary Islands. The bacterial isolates were characterized and identified as Pantoea agglomerans. Pathogenicity tests conducted on onion bulbs and mango plants confirmed that P. agglomerans strains isolated from mango trees are a new etiological agent of a bacterial necrotic disease in the Canary Islands. Pathogenicity plasmids of the pPATH family have been previously reported in P. agglomerans. The majority of putatively pathogenic (n = 23) and pathogenic (n = 4) P. agglomerans strains isolated from mango trees harbored four plasmids, one of which was close in size to the 135-kb pPATH pathogenicity plasmid. The analysis of the presence of two major genes in pPATH plasmids (repA and hrpJ) was undertaken in P. agglomerans strains isolated from mango trees. The hrpJ gene was detected in the 140-kb plasmid of pathogenic P. agglomerans strains isolated from mango trees but it showed differences in nucleotide sequences compared with other pathogenic strains. In contrast, the repA gene was not detected in any of the putatively pathogenic and pathogenic P. agglomerans strains isolated from mango trees. Finally, genetic characterization and phylogenetic analysis using the hrpJ gene and the housekeeping genes gyrB and rpoB showed that almost all P. agglomerans strains that were putatively pathogenic and pathogenic on mango trees clustered together, forming a differentiated phylogroup with respect to the other pathogenic P. agglomerans strains described from other hosts.

Antimicrobial resistance and molecular characterization of Pantoea agglomerans isolated from rainbow trout (Oncorhynchus mykiss) fry.

Aquaculture has become an important candidate as an animal protein source through its growth over the last decade. Based upon a report from the Food and Agriculture Organization of the United Nations, it is the fastest growing sector of the food industry, yet the pathogenicity of many biological agents involved in aquaculture is still unknown. In this study, we isolated Pantoe agglomerans from diseased rainbow trout on several occasions and also attempted to determine their phenotypic and genotypic characteristics, including antimicrobial resistance, of four bacterial isolates. In the present study, P. agglomerans was isolated from diseased rainbow trout as a pathogenic agent. The identification of the P. agglomerans isolates from the rainbow trout was performed through biochemical tests and 16S rRNA sequence analysis. These isolates were predominately biochemically homogeneous, although some features were different, such as seen in methyl-red, mannose and lipase activity tests. All four studied isolates were identified as 99% identical to P. agglomerans based on sequence analysis. The isolates were compared through a phylogenetic analysis with P. agglomerans sequences recovered from 16 other countries and accessed from the GenBank database. All isolates in our study were at least 98.2% similar to sequences from GenBank. Furthermore, the phenotypic antimicrobial susceptibility of the isolates in this study was analyzed through both disc diffusion and broth micro dilution minimum inhibitory concentration (MIC) tests. Although there were some differences between two phenotypic antimicrobial tests, all studied isolates were found susceptible to different antimicrobials. In addition genotypic antimicrobial resistance characteristics were assessed by the presence of antimicrobial resistance genes (ARGs), in which qnrS and sul2 were detected for the first time in P. agglomerans.

Revealing the inventory of type III effectors in Pantoea agglomerans gall-forming pathovars using draft genome sequences and a machine-learning approach.

Pantoea agglomerans, a widespread epiphytic bacterium, has evolved into a hypersensitive response and pathogenicity (hrp)-dependent and host-specific gall-forming pathogen by the acquisition of a pathogenicity plasmid containing a type III secretion system (T3SS) and its effectors (T3Es). Pantoea agglomerans pv. betae (Pab) elicits galls on beet (Beta vulgaris) and gypsophila (Gypsophila paniculata), whereas P. agglomerans pv. gypsophilae (Pag) incites galls on gypsophila and a hypersensitive response (HR) on beet. Draft genome sequences were generated and employed in combination with a machine-learning approach and a translocation assay into beet roots to identify the pools of T3Es in the two pathovars. The genomes of the sequenced Pab4188 and Pag824-1 strains have a similar size (∼5 MB) and GC content (∼55%). Mutational analysis revealed that, in Pab4188, eight T3Es (HsvB, HsvG, PseB, DspA/E, HopAY1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on beet and gypsophila. In Pag824-1, nine T3Es (HsvG, HsvB, PthG, DspA/E, HopAY1, HopD1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on gypsophila, whereas the PthG effector triggers HR on beet. HsvB, HsvG, PthG and PseB appear to endow pathovar specificities to Pab and Pag, and no homologous T3Es were identified for these proteins in other phytopathogenic bacteria. Conversely, the remaining T3Es contribute to the virulence of both pathovars, and homologous T3Es were found in other phytopathogenic bacteria. Remarkably, HsvG and HsvB, which act as host-specific transcription factors, displayed the largest contribution to disease development.

Clinical and microbiological characteristics of Pantoea agglomerans infection in children.

Pantoea agglomerans is an environmental Gram-negative bacterium that rarely is responsible for the infections in humans but it is often a causative factor of a number of occupational diseases. This study evaluated the clinical and microbiological characteristics and pathogenicity of P. agglomerans in children. We retrospectively reviewed microbiological test results for all children (1 month old to 18 years old) who were admitted to our pediatric hospital between January 2000 to June 2015 and had positive clinical specimen cultures for P. agglomerans. Isolates were identified using conventional tests and the BBL Crystal E/NF ID or MALDI-TOF MS systems. Antibiotic susceptibilities were evaluated using the Kirby-Bauer disc diffusion method. We identified fifteen positive cultures from 14 patients with confirmed infections. The positive specimens included pus, urine, tracheal aspirate, blood, and central venous line samples that yielded P. agglomerans. The median patient age was 8.8 years (range: 1.5 months to 16.5 years), and all patients had underlying comorbidities. Five patients had medical devices, and two devices were removed. The most common P. agglomerans infections involved wound infections (35.7%), pneumonia (21.4%), and urinary tract infections (21.4%). Three patients had concomitant infections (Enterococcus faecium, Pseudomonas aeruginosa, and Aspergillus fumigatus). Five patients had anemia. Three patients (21.4%) died, and all three had carbapenem-resistant P. agglomerans that was detected after the first week of hospitalization; two cases involved pneumonia, which was ineffectively treated. P. agglomerans infections may be life-threatening, especially in young patients with pneumonia. Hospital-acquired P. agglomerans may have different pathogenicity and clinical features, compared to community-acquired P. agglomerans, although further studies are needed to understand the drug-resistance patterns in this bacterium.

Pantoea Species Bacteremia in a Child With Sickle Cell Disease: Looking for a Culprit.

Pantoea agglomerans has been classically associated with cellulitis or synovitis secondary to penetrating trauma by vegetation. It is an infrequent cause of systemic infections. We describe the case of a 5-year-old girl with sickle cell disease with P. agglomerans bacteremia and review its potential causes.

Swimming and twitching motility are essential for attachment and virulence of Pantoea ananatis in onion seedlings.

Pantoea ananatis is a widespread phytopathogen with a broad host range. Despite its ability to infect economically important crops, such as maize, rice and onion, relatively little is known about how this bacterium infects and colonizes host tissue or spreads within and between hosts. To study the role of motility in pathogenicity, we analysed both swimming and twitching motility in P. ananatis LMG 20103. Genetic recombineering was used to construct four mutants affected in motility. Two flagellar mutants were disrupted in the flgK and motA genes, required for flagellar assembly and flagellar rotation, respectively. Similarly, two twitching motility mutants were generated, impaired in the structure (pilA) and functioning (pilT) of the type IV pili. The role of swimming and twitching motility during the infection cycle of P. ananatis in onion seedlings was determined by comparing the mutant- and wild-type strains using several in vitro and in planta assays. From the results obtained, it was evident that flagella aid P. ananatis in locating and attaching to onion leaf surfaces, as well as in pathogenicity, whereas twitching motility is instrumental in the spread of the bacteria on the surface once attachment has occurred. Both swimming and twitching motility contribute towards the ability of P. ananatis to cause disease in onions.

Characterization of two LuxI/R homologs in Pantoea ananatis LMG 2665T.

Quorum sensing (QS) plays an important role in the regulation of bacteria-host interactions and ecological fitness in many bacteria. In this study, 2 luxI/R homologs, namely eanI/eanR and rhlI/rhlR, were identified in the genome sequence of Pantoea ananatis LMG 2665T. To determine a role for these luxI/R homologs in pathogenicity and biofilm formation, mutant bacterial strains lacking either eanI/R or rhlI/R and both of these homologs were generated. The results indicated that both the RhlI/R and EanI/R systems are required for pathogenicity and biofilm formation in strain LMG 2665T. This is the first study to characterize the biological significance of the RhlI/R QS system in P. ananatis.

Pantoea agglomerans: a mysterious bacterium of evil and good. Part III. Deleterious effects: infections of humans, animals and plants.

Pantoea agglomerans, a bacterium associated with plants, is not an obligate infectious agent in humans. However, it could be a cause of opportunistic human infections, mostly by wound infection with plant material, or as a hospital-acquired infection, mostly in immunocompromised individuals. Wound infection with P. agglomerans usually follow piercing or laceration of skin with a plant thorn, wooden splinter or other plant material and subsequent inoculation of the plant-residing bacteria, mostly during performing of agricultural occupations and gardening, or children playing. Septic arthritis or synovitis appears as a common clinical outcome of exogenous infection with P. agglomerans, others include endophthalmitis, periostitis, endocarditis and osteomyelitis. Another major reason for clinical infection with P. agglomerans is exposure of hospitalized, often immunodeficient individuals to medical equipment or fluids contaminated with this bacterium. Epidemics of nosocomial septicemia with fatal cases have been described in several countries, both in adult and paediatric patients. In most cases, however, the clinical course of the hospital-acquired disease was mild and application of the proper antibiotic treatment led to full recovery. Compared to humans, there are only few reports on infectious diseases caused by Pantoea agglomerans in vertebrate animals. This species has been identified as a possible cause of equine abortion and placentitis and a haemorrhagic disease in dolphin fish (Coryphaena hippurus). P. agglomerans strains occur commonly, usually as symbionts, in insects and other arthropods. Pantoea agglomerans usually occurs in plants as an epi- or endophytic symbiont, often as mutualist. Nevertheless, this species has also also been identified as a cause of diseases in a range of cultivable plants, such as cotton, sweet onion, rice, maize, sorghum, bamboo, walnut, an ornamental plant called Chinese taro (Alocasia cucullata), and a grass called onion couch (Arrhenatherum elatius). Some plant-pathogenic strains of P. agglomerans are tumourigenic, inducing gall formation on table beet, an ornamental plant gypsophila (Gypsophila paniculata), wisteria, Douglas-fir and cranberry. Recently, a Pantoea species closely related to P. agglomerans has been identified as a cause of bacterial blight disease in the edible mushroom Pleurotus eryngii cultivated in China. The genetically governed determinants of plant pathogenicity in Pantoea agglomerans include such mechanisms as the hypersensitive response and pathogenicity (hrp) system, phytohormones, the quorum-sensing (QS) feedback system and type III secretion system (T3SS) injecting the effector proteins into the cytosol of a plant cell.

Perturbation of maize phenylpropanoid metabolism by an AvrE family type III effector from Pantoea stewartii.

AvrE family type III effector proteins share the ability to suppress host defenses, induce disease-associated cell death, and promote bacterial growth. However, despite widespread contributions to numerous bacterial diseases in agriculturally important plants, the mode of action of these effectors remains largely unknown. WtsE is an AvrE family member required for the ability of Pantoea stewartii ssp. stewartii (Pnss) to proliferate efficiently and cause wilt and leaf blight symptoms in maize (Zea mays) plants. Notably, when WtsE is delivered by a heterologous system into the leaf cells of susceptible maize seedlings, it alone produces water-soaked disease symptoms reminiscent of those produced by Pnss. Thus, WtsE is a pathogenicity and virulence factor in maize, and an Escherichia coli heterologous delivery system can be used to study the activity of WtsE in isolation from other factors produced by Pnss. Transcriptional profiling of maize revealed the effects of WtsE, including induction of genes involved in secondary metabolism and suppression of genes involved in photosynthesis. Targeted metabolite quantification revealed that WtsE perturbs maize metabolism, including the induction of coumaroyl tyramine. The ability of mutant WtsE derivatives to elicit transcriptional and metabolic changes in susceptible maize seedlings correlated with their ability to promote disease. Furthermore, chemical inhibitors that block metabolic flux into the phenylpropanoid pathways targeted by WtsE also disrupted the pathogenicity and virulence activity of WtsE. While numerous metabolites produced downstream of the shikimate pathway are known to promote plant defense, our results indicate that misregulated induction of phenylpropanoid metabolism also can be used to promote pathogen virulence.

Pantoea ananatis Utilizes a Type VI Secretion System for Pathogenesis and Bacterial Competition.

Type VI secretion systems (T6SSs) are a class of macromolecular machines that are recognized as an important virulence mechanism in several gram-negative bacteria. The genome of Pantoea ananatis LMG 2665(T), a pathogen of pineapple fruit and onion plants, carries two gene clusters whose predicted products have homology with T6SS-associated gene products from other bacteria. Nothing is known regarding the role of these T6SS-1 and T6SS-3 gene clusters in the biology of P. ananatis. Here, we present evidence that T6SS-1 plays an important role in the pathogenicity of P. ananatis LMG 2665(T) in onion plants, while a strain lacking T6SS-3 remains as pathogenic as the wild-type strain. We also investigated the role of the T6SS-1 system in bacterial competition, the results of which indicated that several bacteria compete less efficiently against wild-type LMG 2665(T) than a strain lacking T6SS-1. Additionally, we demonstrated that these phenotypes of strain LMG 2665(T) were reliant on the core T6SS products TssA and TssD (Hcp), thus indicating that the T6SS-1 gene cluster encodes a functioning T6SS. Collectively, our data provide the first evidence demonstrating that the T6SS-1 system is a virulence determinant of P. ananatis LMG 2665(T) and plays a role in bacterial competition.

[THE SENSITIVITY OF PHYTOPATHOGENIC BACTERIA TO STREPTOMYCIN UNDER THE INFLUENCE OF PESTICIDES].

The results of the streptomycin sensitivity changes of phytopathogenic Pseudomonas syringae and Xanthomonas translucens bacteria under the action of pesticides are pre- sented. It is demonstrated that phytopathogenic strains show greater changes of strepto- mycin sensitivity compared to epiphytic Pantoea agglomerans strain under the pesticides influence. Granstar herbicide, Tviks and Alpha Super insecticides increase the number of streptomycin resistant cells of Xanthomonas translucens 3164, P syringae pv. syringae YKM B-1027 and P syringae pv. atrofaciens YKM B-1011. This fact indicates mutagenic action of these pesticides against researched phytopathogenic bacteria.

An unusual cause of peritonitis in peritoneal dialysis patients: Pantoea agglomerans.

Peritonitis is a serious infection and early diagnosis and treatment is mandatory. A variety of microorganisms are identified in these cases and during recent years a new one was included, Pantoea agglomerans. In this case report, a female patient on continuous ambulatory peritoneal dialysis therapy with a peritonitis episode caused by this organism is described. The source of infection was thought to be due to contact of catheter with non-sterile surfaces. In microbiologic culture, this organism was identified and the patient successfully treated with a three week course of gentamicin therapy. The number of reported cases with this organism has increased in last years and various infection localizations and clinical progress patterns have been identified. In peritoneal dialysis patients presenting with peritonitis, this organism must be kept in mind.