Genomic analysis of eight native plasmids of the phytopathogen Pseudomonas syringae

Comunicación a conferenciaThe pPT23A family of plasmids (PFPs) appears to be indigenous to the plant pathogen Pseudomonas syringae and these plasmids are widely distributed and widely transferred among pathovars of P. syringae and related species. PFPs are sources of accessory genes for their hosts that can include genes important for virulence and epiphytic colonization of plant leaf surfaces. Further understanding of the evolution of the pPT23A plasmid family and the role of these plasmids in P. syringae biology and pathogenesis, requires the determination and analysis of additional complete, closed plasmid genome sequences. Therefore, our main objective was to obtain complete genome sequences from PFPs from three different P. syringae pathovars and perform a comparative genomic analysis. In this work plasmid DNA isolation, purification by CsCl-EtBr gradients, and sequencing using 454 platform, were used to obtain the complete sequence of P. syringae plasmids. Different bioinformatic tools were used to analyze the plasmid synteny, to identify virulence genes (i.e. type 3 effectors) and to unravel the evolutionary history of PFPs. Our sequence analysis revealed that PFPs from P. syringae encode suites of accessory genes that are selected at different levels (universal, interpathovar and intrapathovar). The conservation of type IVSS encoding conjugation functions also contributes to the distribution of these plasmids within P. syringae populations. Thus, this study contributes to unravel the genetic basis of the role of PFPs in different P. syringae lifestyles.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Isolation, characterization and selection of bacterial isolates from a suppressive soil with beneficial traits to plants

Backgrounds This study focused on the characterization and selection of bacterial strains obtained from a suppressive soil displaying antifungal activity against the soilborne phytopathogenic fungi Rosellinia necatrix. Bacterial profile from this suppressive soil were first obtained by 16S rRNA gene sequencing, revealing a significant increase in the bacterial class Gammaproteobacteria, especially in some antagonistic representatives of Pseudomonas spp. Objectives To obtain and characterize a collection of 246 bacterial isolates obtained from this suppressive soil, in order to identify new strains with antifungal activity against fungal phytopathogens. Methods To obtain the bacterial collection, we performed an isolation on a selective medium for Pseudomonas-like microorganisms. Further characterization tests were used in order to analyse the bacterial collection, including identification of the general metabolic profile of glucose, the profiling of antifungals produced, including both the putative production of antifungal compounds and lytic exoenzymes, and the evaluation of traits related with beneficial effects on plants. Conclusions A final selection of representative strains resulted in antifungal isolates belonging to the genus Pseudomonas, but also some representatives of the genera Serratia and Stenotrophomonas. These selected strains were tested for plant protection by an in vivo experiment using avocado and wheat plants challenged by the pathogen R. necatrix, showing all of them an antifungal ability and plant disease protection. Pseudomonas-like strains isolated from suppressive soils constitute an excellent source for novel microbial biocontrol agents against soilborne fungal pathogens. This work was supported by grant AGL2014-52518-C2-1-R. Carmen Vida and Sandra Tienda are supported by a PhD fellowship from the FPI program of the Spanish Government.This work was supported by grant AGL2014-52518-C2-1-R. Carmen Vida and Sandra Tienda are supported by a PhD fellowship from the FPI program of the Spanish Government; Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Comparative genomic analysis of native pseudomonas syringae plasmids belonging to the ppt23 a family reveals their role in p. Syringae epiphytic and pathogenic lifestyles

Backgrounds The pPT23A family of plasmids (PFPs) appears to be indigenous to the plant pathogen Pseudomonas syringae and these plasmids are widely distributed and widely transferred among pathovars of P. syringae and related species. PFPs are sources of accessory genes for their hosts that can include genes important for virulence and epiphytic colonization of plant leaf surfaces. Objectives Further understanding of the evolution of the pPT23A plasmid family and the role of these plasmids in P. syringae biology and pathogenesis, requires the determination and analysis of additional complete, closed plasmid genome sequences. Therefore, our main objective was to obtain complete genome sequences of PFPs from three different P. syringae pathovars and perform a comprehensive comparative genomic analysis. Methods In this work plasmid DNA isolation, purification by CsCl-EtBr gradients, and sequencing using 454 platform, were carried out to obtain the complete sequence of P. syringae plasmids. Different bioinformatic tools were used to analyze the plasmid synteny, to identify virulence genes (i.e. type 3 effectors) and to unravel the evolutionary history of PFPs. Conclusions Our sequence analysis revealed that PFPs from P. syringae encode suites of accessory genes that are selected at different levels (universal, interpathovar and intrapathovar). The conservation of type IVSS encoding conjugation functions also contributes to the distribution of these plasmids within P. syringae populations. Thus, this study contributes to unravel the genetic bases of the role of PFPs in different P. syringae lifestyles. This work was supported by grants Proyecto de Excelencia, Junta de Andalucía (P07-AGR-02471; P12-AGR-1473) and by Michigan State University AgBioResearch.This work was supported by grants Proyecto de Excelencia, Junta de Andalucía (P07-AGR-02471; P12-AGR-1473) and by Michigan State University AgBioResearch; Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Genome Sequence of Paracoccus sp. JM45, a Bacterial Strain Isolated from a Marine Sponge with a Dual Quorum Sensing Inhibition Activity.

The draft genome sequence of Paracoccus sp. strain JM45, isolated from a marine sponge harvested off the west coast of Ireland, is reported here. Quorum sensing and quorum sensing inhibition activities have been reported recently for this bacterium, and genomic analysis supports its potential use for novel therapeutic development

Arsenic, fluoride and other trace elements in the Argentina Pampean plain

The contents of arsenic (As), fluoride (F) and other trace elements (B, Cd, Cr, Cu, Fe, Mn, Ni, Pb, V, Zn, Ba, Si and Sr) have been determined in groundwater samples from the Langueyú creek basin, in the Argentina Pampean plain. This research aims to establish the baseline concentration and geographical distribution of trace elements in this basin. This aim has particular interest to public health in the city of Tandil where groundwater is the principal source of water for human supply. The baseline concentrations of elements in the Langueyú creek basin are in good agreement with published data from other locations of the Pampean aquifer. The arsenic limit of 10mg/l, established as provisional limit by the World Health Organization (WHO), was exceeded in 78% of the sampled wells, with As concentration increasing in the direction of groundwater flow. Concentrations of B, Cd, Cu, Cr, Fe, Mn, Ni, Pb and Zn regulated by the Argentinian Food Code (CAA) do not exceed the maximum limit for drinking water, although concentrations of Ni, Zn or Pb peaked up at some wells, probably due to pipeline corrosion. The strong correlation observed between As, F, V, Cr and B has been related to their anionic character at the groundwater natural alkaline pH that is likely associated with similar mobilization (adsorption/desorption) processes. Worst consequences for human health have arisen in areas with the highest arsenic concentration in drinking water. The conclusions of this study contribute to understand the provenance and mobilization processes of some trace elements in groundwater. It enables the decision making regarding the public health priorities and the technological treatments of water resources in urban and rural areas

Coumarin: a novel player in microbial quorum sensing and biofilm formation inhibition

Antibiotic resistance is a growing threat worldwide, causing serious problems in the treatment of microbial infections. The discovery and development of new drugs is urgently needed to overcome this problem which has greatly undermined the clinical effectiveness of conventional antibiotics. An intricate cell-cell communication system termed quorum sensing (QS) and the coordinated multicellular behaviour of biofilm formation have both been identified as promising targets for the treatment and clinical management of microbial infections. QS systems allow bacteria to adapt rapidly to harsh conditions, and are known to promote the formation of antibiotic tolerant biofilm communities. It is well known that biofilm is a recalcitrant mode of growth and it also increases bacterial resistance to conventional antibiotics. The pharmacological properties of coumarins have been well described, and these have included several that possess antimicrobial properties. More recently, reports have highlighted the potential role of coumarins as alternative therapeutic strategies based on their ability to block the QS signalling systems and to inhibit the formation of biofilms in clinically relevant pathogens. In addition to human infections, coumarins have also been found to be effective in controlling plant pathogens, infections in aquaculture, food spoilage and in reducing biofouling caused by eukaryotic organisms. Thus, the coumarin class of small molecule natural product are emerging as a promising strategy to combat bacterial infections in the new era of antimicrobial resistance

A novel gene cluster involved in high tolerance to copper compounds has been detected in Pseudomonas syringae pv. syringae

Pseudomonas syringae pv. syringae (Pss), is the most polyphagous bacterium of the P. syringae complex, based on its wide host range, that severely affects woody and herbaceous plants. Pss has been described as the causal agent of bacterial apical necrosis (BAN) on mango crops, an important threat that drastically reduces the mango fruit yield in the Mediterranean region. Copper compounds have traditionally been used as standard bactericides to manage woody plant diseases, being the spraying of copper compound known as Bordeaux mixture (BM), the conventional treatment for controlling BAN disease. The extensive use of copper compounds can lead to many problems, and among them, the emergence of copper-resistant strains is one of the most serious. Pss strains isolated from mango trees harbour at least, two different genetic structures (copABCD and copABCD modified) encoded by plasmids that are involved in copper detoxification. Determination of the minimum inhibitory concentration of copper, together with a phylogenetic distribution analysis performed in a collection of Pss strains isolated from mango trees revealed a phylogenetic group of Pss strongly associated with high levels of copper resistance. Genome mining and functional characterization of one Pss strain present in this group have uncovered a novel gene cluster of chromosomal location that could be playing a major role in high levels of copper resistance during the epiphytic survival of Pss on mango tree surfaces.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Análisis comparativo de dos colecciones de pseudo monas syringae pv. syringae asociadas a mango

Pseudomonas syringae es una bacteria ubicua a nivel ambiental. Presenta una taxonomía compleja en la que se han descrito más de 60 patovares que causan enfermedad en muchas especies de plantas diferentes. Entre los patovares más importantes se encuentra el pv. syringae debido a su amplio rango de hospedador, y al arsenal de mecanismos de virulencia que posee. A principios de los años 90, se describió a Pseudomonas syringae pv. syringae (Pss) como el agente causal de la necrosis apical del mango (NAM). Pss aisladas de mango muestran características importantes para su biología, tanto en sus mecanismos de virulencia como para el fitness epifítico. En este trabajo, se plantea conocer en profundidad cómo han evolucionado las poblaciones de Pss a lo largo del tiempo sobre el mangoUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Este trabajo ha sido financiado por ayuda de la Consejería de Economía, Innovación y Ciencia, Junta de Andalucía (Pl2-AGR-1473), cofinanciados con fondos FEDER (UE

Aislamiento de cepas de Pseudomonas syringae pv. syringae asociadas a mango para su uso en análisis evolutivos y epidemiológicos

Comunicación oralLa necrosis apical del mango (NAM) es una enfermedad que se ha observado en el litoral andaluz desde la implantación de este cultivo, y cuyo agente causal es la bacteria Pseudomonas syringae pv. syringae (Pss). Esta enfermedad aparece principalmente asociada a climas con inviernos frescos y húmedos, tal y como ocurre en la cuenca mediterránea, donde, además de en España, se ha descrito en otros países (Israel, Portugal, Italia, Egipto), así como en otras áreas de cultivo con clima similar, como el Noroeste de Australia. Las cepas de Pss aisladas de mango muestran características importantes para su biología, tanto a nivel de virulencia como de fitness epifítico. En trabajos previos se han descrito diferentes genes implicados en el desarrollo de los síntomas de la enfermedad, así como en aumentar la capacidad de la bacteria de sobrevivir y persistir sobre el tejido vegetal; el operón mbo implicado en la producción de mangotoxina, los genes copABCD o cusCBA, implicados en la capacidad de resistencia al cobre, o el operón wss, responsable de la producción de celulosa. Por otro lado, gracias a análisis filogenéticos, se ha podido agrupar a todas las cepas de Pss aisladas de mango y productoras de mangotoxinas en el filotipo I. El objetivo de este trabajo es el análisis fenético comparado de cepas de Pss aisladas de mango de las diferentes zonas de estudio (España, Portugal, Italia, Australia), antes del año 2000 (colección I) y disponibles en nuestro laboratorio, con aislamientos actuales (2016-2018, Colección II). Este estudio se ha iniciado con la caracterización y selección previa de cepas representativas de cada una de las colecciones, para iniciar un abordaje en detalle y comparación de los diferentes atributos.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Contribution of mangotoxin to the virulence and epiphytic fitness of Pseudomonas syringae pv. syringae

Mangotoxin is an antimetabolite toxin that inhibits ornithine acetyl transferase, a key enzyme in the biosynthetic pathway of ornithine and arginine and recently reported in strains of Pseudomonas syringae pv. syringae (Pss) isolated from mango. Since symptoms on mango tissues are very difficult to reproduce, in this study the role of mangotoxin in Pss virulence was addressed by analyzing the in planta growth and development of disease symptoms on tomato leaflets. Inoculation experiments were carried out following several procedures using the wild-type strain Pss UMAF0158, two Tn5-mutant derivative strains defective in mangotoxin production, and their complemented derivative strains in which mangotoxin production is restored. The ability of the mangotoxin-defective mutants to grow in planta was similar, and their epiphytic survival on the tomato leaf surface identical to the wild-type and complemented strains. However, both the disease index data of incidence and the severity of necrotic symptoms indicated that mangotoxin-defective mutants were less virulent, indicating that mangotoxin is a virulence factor. Furthermore, competition experiments showed that the survival values of the wild-type strain were slightly but significantly higher than those of the mangotoxin-defective mutants, suggesting that mangotoxin production would improve the epiphytic fitness of Pss. [Int Microbiol 2009; 12(2):87-95