RESUMO
Pathogen introductions have led to numerous disease outbreaks in naive regions of the globe. The plant pathogen Xylella fastidiosa has been associated with various recent epidemics in Europe affecting agricultural crops, such as almond, grapevine, and olive, but also endemic species occurring in natural forest landscapes and ornamental plants. We compared whole-genome sequences of X. fastidiosa subspecies multiplex from America and strains associated with recent outbreaks in southern Europe to infer their likely origins and paths of introduction within and between the two continents. Phylogenetic analyses indicated multiple introductions of X. fastidiosa subspecies multiplex into Italy, Spain, and France, most of which emerged from a clade with limited genetic diversity with a likely origin in California, USA. The limited genetic diversity observed in X. fastidiosa subspecies multiplex strains originating from California is likely due to the clade itself being an introduction from X. fastidiosa subspecies multiplex populations in the southeastern United States, where this subspecies is most likely endemic. Despite the genetic diversity found in some areas in Europe, there was no clear evidence of recombination occurring among introduced X. fastidiosa strains in Europe. Sequence type taxonomy, based on multilocus sequence typing (MLST), was shown, at least in one case, to not lead to monophyletic clades of this pathogen; whole-genome sequence data were more informative in resolving the history of introductions than MLST data. Although additional data are necessary to carefully tease out the paths of these recent dispersal events, our results indicate that whole-genome sequence data should be considered when developing management strategies for X. fastidiosa outbreaks.IMPORTANCEXylella fastidiosa is an economically important plant-pathogenic bacterium that has emerged as a pathogen of global importance associated with a devastating epidemic in olive trees in Italy associated with X. fastidiosa subspecies pauca and other outbreaks in Europe, such as X. fastidiosa subspecies fastidiosa and X. fastidiosa subspecies multiplex in Spain and X. fastidiosa subspecies multiplex in France. We present evidence of multiple introductions of X. fastidiosa subspecies multiplex, likely from the United States, into Spain, Italy, and France. These introductions illustrate the risks associated with the commercial trade of plant material at global scales and the need to develop effective policy to limit the likelihood of pathogen pollution into naive regions. Our study demonstrates the need to utilize whole-genome sequence data to study X. fastidiosa introductions at outbreak stages, since a limited number of genetic markers does not provide sufficient phylogenetic resolution to determine dispersal paths or relationships among strains that are of biological and quarantine relevance.
Assuntos
Genoma Bacteriano , Doenças das Plantas/microbiologia , Xylella/genética , Brasil , Europa (Continente) , Espécies Introduzidas , Sequenciamento Completo do GenomaRESUMO
Xylella fastidiosa is a plant-pathogenic bacterium recently introduced in Europe that is causing decline in olive trees in the South of Italy. Genetic studies have consistently shown that the bacterial genotype recovered from infected olive trees belongs to the sequence type ST53 within subspecies pauca. This genotype, ST53, has also been reported to occur in Costa Rica. The ancestry of ST53 was recently clarified, showing it contains alleles that are monophyletic with those of subsp. pauca in South America. To more robustly determine the phylogenetic placement of ST53 within X. fastidiosa, we performed a comparative analysis based on single nucleotide polymorphisms (SNPs) and the study of the pan-genome of the 27 currently public available whole genome sequences of X. fastidiosa. The resulting maximum-parsimony and maximum likelihood trees constructed using the SNPs and the pan-genome analysis are consistent with previously described X. fastidiosa taxonomy, distinguishing the subsp. fastidiosa, multiplex, pauca, sandyi, and morus. Within the subsp. pauca, the Italian and three Costa Rican isolates, all belonging to ST53, formed a compact phylotype in a clade divergent from the South American pauca isolates, also distinct from the recently described coffee isolate CFBP8072 imported into Europe from Ecuador. These findings were also supported by the gene characterization of a conjugative plasmid shared by all the four ST53 isolates. Furthermore, isolates of the ST53 clade possess an exclusive locus encoding a putative ATP-binding protein belonging to the family of histidine kinase-like ATPase gene, which is not present in isolates from the subspecies multiplex, sandyi, and pauca, but was detected in ST21 isolates of the subspecies fastidiosa from Costa Rica. The clustering and distinctiveness of the ST53 isolates supports the hypothesis of their common origin, and the limited genetic diversity among these isolates suggests this is an emerging clade within subsp. pauca.
Assuntos
Genoma Bacteriano , Estudo de Associação Genômica Ampla , Genótipo , Xylella/genética , Costa Rica , DNA Bacteriano/genética , Regulação Bacteriana da Expressão Gênica , Itália , Filogenia , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Aiming to combine the flexibility of Brucella lumazine synthase (BLS) to adapt different protein domains in a decameric structure and the capacity of BLS and flagellin to enhance the immunogenicity of peptides that are linked to their structure, we generated a chimeric protein (BLS-FliC131) by fusing flagellin from Salmonella in the N-termini of BLS. The obtained protein was recognized by anti-flagellin and anti-BLS antibodies, keeping the oligomerization capacity of BLS, without affecting the folding of the monomeric protein components determined by circular dichroism. Furthermore, the thermal stability of each fusion partner is conserved, indicating that the interactions that participate in its folding are not affected by the genetic fusion. Besides, either in vitro or in vivo using TLR5-deficient animals we could determine that BLS-FliC131 retains the capacity of triggering TLR5. The humoral response against BLS elicited by BLS-FliC131 was stronger than the one elicited by equimolar amounts of BLS + FliC. Since BLS scaffold allows the generation of hetero-decameric structures, we expect that flagellin oligomerization on this protein scaffold will generate a new vaccine platform with enhanced capacity to activate immune responses.
Assuntos
Brucella , Flagelina , Complexos Multienzimáticos , Proteínas Recombinantes de Fusão , Salmonella typhimurium , Animais , Brucella/enzimologia , Brucella/genética , Brucella/imunologia , Células CACO-2 , Feminino , Flagelina/biossíntese , Flagelina/genética , Flagelina/imunologia , Humanos , Imunidade Humoral , Camundongos , Camundongos Knockout , Complexos Multienzimáticos/biossíntese , Complexos Multienzimáticos/genética , Complexos Multienzimáticos/imunologia , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Salmonella typhimurium/genética , Salmonella typhimurium/imunologia , Salmonella typhimurium/metabolismo , Receptor 5 Toll-Like/genética , Receptor 5 Toll-Like/imunologiaRESUMO
Ante la acelerada pérdida de biodiversidad causada por actividades humanas, las áreas protegidas (APs) constituyen la principal respuesta para mitigar esta crisis. La Reserva Natural Iberá (RNI) incluye 13000 km2 de territorios privados y fiscales, que contienen humedales y tierras altas con elevada biodiversidad, endemismos e importantes poblaciones de especies amenazadas. Para implementar su conservación, a finales del siglo pasado se definieron cinco Unidades de Conservación (UC) priorizadas en infraestructura y guardaparques, seleccionadas mediante criterios pobremente explicitados. En este trabajo definimos áreas prioritarias para la conservación (APC) usando los patrones de distribución de los reptiles en la RNI, para evaluar la eficiencia de las UC instauradas y detectar otras posibles áreas a ser consideradas en gestiones de conservación. La RNI se dividió en 28 celdas de 0,25° de latitud-longitud y mediante muestreos de campo y revisión de colecciones se obtuvieron 1 482 registros de 71 especies de reptiles computándose su presencia/ausencia en las celdas. Se calculó por celda riqueza específica e Índice Combinado de Biodiversidad (ICB) (que incluye rareza y grado de amenaza). Las APC fueron definidas mediante una búsqueda exacta (Complementariedad), obteniendo el mínimo conjunto de áreas que contuvieran a todas las especies y valores del ICB, lo que se denomina "eficiencia máxima". La superficie mínima necesaria que representa a todas las especies es de diez celdas (36%) y sólo dos incluyen UC actuales. Se necesitan adicionar ocho celdas para cubrir los vacíos de conservación de reptiles, particularmente en el norte de la RNI. Sobre 20 especies amenazadas o insuficientemente conocidas, 12 (60%) no fueron registradas en celdas con UC. Proponemos UC adicionales para efectivizar la protección de todos los reptiles e incluir especies amenazadas.(AU)
Owing to accelerated biodiversity loss caused by human activities, the protected areas (PAs) are the main response to mitigate this crisis. The Iberá Natural Reserve (INR) includes 13000 km2 of private and state lands, which contain wetland and upland areas with high biodiversity, endemism and significant populations of threatened species. In order to implement its conservation, late last century, five Units of Conservation (UC) were defined and prioritized due to infrastructure and park-rangers, although poorly selected by explicit criteria. We defined priority areas for conservation (APCs) using the distribution patterns of reptiles in the INR, in order to evaluate the efficiency of the five UC located within its boundaries and detect other possible areas to be considered in conservation efforts. The INR was divided into 28 quadrat-cells of 0.25° latitude-longitude and through field samplings and revision of collections we obtained 1482 records of 71 species of reptiles, computed their presence / absence in the cells. We estimated species richness and Combined Biodiversity Index (including rarity and degree of threat) by cell. APCs were defined by an exact search (complementarity index), obtaining the minimum set of areas containing higher values of the indexes used, which is called "maximum efficiency". The minimum area required to represent all species was ten cells (36%) and only two of them includes current UC. Add eight cells are needed to fill conservation gaps of reptiles, particularly in the north of the INR. About 20 threatened or poorly known species, 12 (60%) were not recorded in cells with UC. We proposed additional UC in order to protect all reptiles and include threatened species.(AU)
Assuntos
Reservas Naturais/análise , Gestão Ambiental/métodos , Eficiência Organizacional , Áreas Protegidas/análise , Répteis , BiodiversidadeRESUMO
Ante la acelerada pérdida de biodiversidad causada por actividades humanas, las áreas protegidas (APs) constituyen la principal respuesta para mitigar esta crisis. La Reserva Natural Iberá (RNI) incluye 13000 km2 de territorios privados y fiscales, que contienen humedales y tierras altas con elevada biodiversidad, endemismos e importantes poblaciones de especies amenazadas. Para implementar su conservación, a finales del siglo pasado se definieron cinco Unidades de Conservación (UC) priorizadas en infraestructura y guardaparques, seleccionadas mediante criterios pobremente explicitados. En este trabajo definimos áreas prioritarias para la conservación (APC) usando los patrones de distribución de los reptiles en la RNI, para evaluar la eficiencia de las UC instauradas y detectar otras posibles áreas a ser consideradas en gestiones de conservación. La RNI se dividió en 28 celdas de 0,25° de latitud-longitud y mediante muestreos de campo y revisión de colecciones se obtuvieron 1 482 registros de 71 especies de reptiles computándose su presencia/ausencia en las celdas. Se calculó por celda riqueza específica e Índice Combinado de Biodiversidad (ICB) (que incluye rareza y grado de amenaza). Las APC fueron definidas mediante una búsqueda exacta (Complementariedad), obteniendo el mínimo conjunto de áreas que contuvieran a todas las especies y valores del ICB, lo que se denomina "eficiencia máxima". La superficie mínima necesaria que representa a todas las especies es de diez celdas (36%) y sólo dos incluyen UC actuales. Se necesitan adicionar ocho celdas para cubrir los vacíos de conservación de reptiles, particularmente en el norte de la RNI. Sobre 20 especies amenazadas o insuficientemente conocidas, 12 (60%) no fueron registradas en celdas con UC. Proponemos UC adicionales para efectivizar la protección de todos los reptiles e incluir especies amenazadas.
Owing to accelerated biodiversity loss caused by human activities, the protected areas (PAs) are the main response to mitigate this crisis. The Iberá Natural Reserve (INR) includes 13000 km2 of private and state lands, which contain wetland and upland areas with high biodiversity, endemism and significant populations of threatened species. In order to implement its conservation, late last century, five Units of Conservation (UC) were defined and prioritized due to infrastructure and park-rangers, although poorly selected by explicit criteria. We defined priority areas for conservation (APCs) using the distribution patterns of reptiles in the INR, in order to evaluate the efficiency of the five UC located within its boundaries and detect other possible areas to be considered in conservation efforts. The INR was divided into 28 quadrat-cells of 0.25° latitude-longitude and through field samplings and revision of collections we obtained 1482 records of 71 species of reptiles, computed their presence / absence in the cells. We estimated species richness and Combined Biodiversity Index (including rarity and degree of threat) by cell. APCs were defined by an exact search (complementarity index), obtaining the minimum set of areas containing higher values of the indexes used, which is called "maximum efficiency". The minimum area required to represent all species was ten cells (36%) and only two of them includes current UC. Add eight cells are needed to fill conservation gaps of reptiles, particularly in the north of the INR. About 20 threatened or poorly known species, 12 (60%) were not recorded in cells with UC. We proposed additional UC in order to protect all reptiles and include threatened species.
Assuntos
Eficiência Organizacional , Gestão Ambiental/métodos , Reservas Naturais/análise , Répteis , Áreas Protegidas/análise , BiodiversidadeRESUMO
RESUMEN Ante la acelerada pérdida de biodiversidad causada por actividades humanas, las áreas protegidas (APs) constituyen la principal respuesta para mitigar esta crisis. La Reserva Natural Iberá (RNI) incluye 13000 km2 de territorios privados y fiscales, que contienen humedales y tierras altas con elevada biodiversidad, endemismos e importantes poblaciones de especies amenazadas. Para implementar su conservación, a finales del siglo pasado se definieron cinco Unidades de Conservación (UC) priorizadas en infraestructura y guardaparques, seleccionadas mediante criterios pobremente explicitados. En este trabajo definimos áreas prioritarias para la conservación (APC) usando los patrones de distribución de los reptiles en la RNI, para evaluar la eficiencia de las UC instauradas y detectar otras posibles áreas a ser consideradas en gestiones de conservación. La RNI se dividió en 28 celdas de 0,25° de latitud-longitud y mediante muestreos de campo y revisión de colecciones se obtuvieron 1 482 registros de 71 especies de reptiles computándose su presencia/ausencia en las celdas. Se calculó por celda riqueza específica e Índice Combinado de Biodiversidad (ICB) (que incluye rareza y grado de amenaza). Las APC fueron definidas mediante una búsqueda exacta (Complementariedad), obteniendo el mínimo conjunto de áreas que contuvieran a todas las especies y valores del ICB, lo que se denomina eficiencia máxima. La superficie mínima necesaria que representa a todas las especies es de diez celdas (36%) y sólo dos incluyen UC actuales. Se necesitan adicionar ocho celdas para cubrir los vacíos de conservación de reptiles, particularmente en el norte de la RNI. Sobre 20 especies amenazadas o insuficientemente conocidas, 12 (60%) no fueron registradas en celdas con UC. Proponemos UC adicionales para efectivizar la protección de todos los reptiles e incluir especies amenazadas.
ABSTRACT Owing to accelerated biodiversity loss caused by human activities, the protected areas (PAs) are the main response to mitigate this crisis. The Iberá Natural Reserve (INR) includes 13000 km2 of private and state lands, which contain wetland and upland areas with high biodiversity, endemism and significant populations of threatened species. In order to implement its conservation, late last century, five Units of Conservation (UC) were defined and prioritized due to infrastructure and park-rangers, although poorly selected by explicit criteria. We defined priority areas for conservation (APCs) using the distribution patterns of reptiles in the INR, in order to evaluate the efficiency of the five UC located within its boundaries and detect other possible areas to be considered in conservation efforts. The INR was divided into 28 quadrat-cells of 0.25° latitude-longitude and through field samplings and revision of collections we obtained 1482 records of 71 species of reptiles, computed their presence / absence in the cells. We estimated species richness and Combined Biodiversity Index (including rarity and degree of threat) by cell. APCs were defined by an exact search (complementarity index), obtaining the minimum set of areas containing higher values of the indexes used, which is called maximum efficiency. The minimum area required to represent all species was ten cells (36%) and only two of them includes current UC. Add eight cells are needed to fill conservation gaps of reptiles, particularly in the north of the INR. About 20 threatened or poorly known species, 12 (60%) were not recorded in cells with UC. We proposed additional UC in order to protect all reptiles and include threatened species.
RESUMO
Two haplotypes of Xylella fastidiosa subsp. pauca (Xfp) that correlated with their host of origin were identified in a collection of 90 isolates infecting citrus and coffee plants in Brazil, based on a single-nucleotide polymorphism in the gyrB sequence. A new single-nucleotide primer extension (SNuPE) protocol was designed for rapid identification of Xfp according to the host source. The protocol proved to be robust for the prediction of the Xfp host source in blind tests using DNA from cultures of the bacterium, infected plants, and insect vectors allowed to feed on Xfp-infected citrus plants. AMOVA and STRUCTURE analyses of microsatellite data separated most Xfp populations on the basis of their host source, indicating that they were genetically distinct. The combined use of the SNaPshot protocol and three previously developed multilocus SSR markers showed that two haplotypes and distinct isolates of Xfp infect citrus and coffee in Brazil and that multiple, genetically different isolates can be present in a single orchard or infect a single tree. This combined approach will be very useful in studies of the epidemiology of Xfp-induced diseases, host specificity of bacterial genotypes, the occurrence of Xfp host jumping, vector feeding habits, etc., in economically important cultivated plants or weed host reservoirs of Xfp in Brazil and elsewhere.
Assuntos
Citrus/microbiologia , Coffea/microbiologia , Variação Genética , Doenças das Plantas/microbiologia , Xylella/genética , Proteínas de Bactérias/genética , Sequência de Bases , Brasil , DNA Girase/genética , Primers do DNA/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Marcadores Genéticos/genética , Genótipo , Interações Hospedeiro-Patógeno , Repetições de Microssatélites/genética , Dados de Sequência Molecular , Polimorfismo de Nucleotídeo Único/genética , Análise de Sequência de DNA , Xylella/classificação , Xylella/isolamento & purificaçãoRESUMO
Reproductive aspects of Dendropsophus sanborni (Anura, Hylidae) in northeastern Argentina. The aim of this investigation was to characterize reproductive aspects of Dendropsophus sanborni (Schmidt, 1944) (Anura, Hylidae) in the northwest of Corrientes province, in northeastern Argentina. The reproductive cycles of males and females, the existence of sexual dimorphism and the males vocalization period were analyzed. Samplings were made from September 2011 to August 2012. Dendropsophus sanborni showed a prolonged reproductive pattern because reproductive activity was observed during most part of the year. Calling males were recorded all months except in July. The spermatogenic cycle can be characterized as potentially continuous. In males, snout-vent length was significantly correlated with mean testicular volume. The species showed sexual dimorphism in body mass, with females larger than males. Post-ovulatory females were found from March to June and most females had oocytes at different stages of maturity, with abundant post-vitellogenic oocytes from August to December.(AU)
El objetivo del presente estudio fue caracterizar aspectos reproductivos de Dendropsophus sanborni (Schmidt, 1944) (Anura, Hylidae) en la región noroeste de la provincia de Corrientes, al Nordeste de Argentina. Para esto, se analizó el ciclo gonadal de machos y hembras, la existencia de dimorfismo sexual y el periodo de vocalización de los machos. Los muestreos se efectuaron desde Septiembre de 2011 hasta Agosto de 2012. Dendropsophus sanborni exhibe un patrón reproductivo prolongado, dado que la actividad reproductiva ocurre durante gran parte del año. Se registraron machos en actividad de vocalización durante todos los meses excepto en julio. El ciclo espermatogénico puede ser caracterizado como potencialmente continuo. En los machos hubo correlación significativa entre la longitud hocico-cloaca y el volumen testicular medio. La especie presentó dimorfismo sexual en relación a la masa corporal, siendo las hembras mayores que los machos. Se encontraron hembras post-ovulatorias desde Marzo a Junio y una mayor proporción de hembras con ovocitos en diferentes estados de maduración, con abundantes ovocitos post-vitelogénicos, en los meses de Agosto a Diciembre.(AU)
Assuntos
Animais , Gônadas , Reprodução/fisiologia , Vocalização Animal , Caracteres SexuaisRESUMO
Reproductive aspects of Dendropsophus sanborni (Anura, Hylidae) in northeastern Argentina. The aim of this investigation was to characterize reproductive aspects of Dendropsophus sanborni (Schmidt, 1944) (Anura, Hylidae) in the northwest of Corrientes province, in northeastern Argentina. The reproductive cycles of males and females, the existence of sexual dimorphism and the males vocalization period were analyzed. Samplings were made from September 2011 to August 2012. Dendropsophus sanborni showed a prolonged reproductive pattern because reproductive activity was observed during most part of the year. Calling males were recorded all months except in July. The spermatogenic cycle can be characterized as potentially continuous. In males, snout-vent length was significantly correlated with mean testicular volume. The species showed sexual dimorphism in body mass, with females larger than males. Post-ovulatory females were found from March to June and most females had oocytes at different stages of maturity, with abundant post-vitellogenic oocytes from August to December.
El objetivo del presente estudio fue caracterizar aspectos reproductivos de Dendropsophus sanborni (Schmidt, 1944) (Anura, Hylidae) en la región noroeste de la provincia de Corrientes, al Nordeste de Argentina. Para esto, se analizó el ciclo gonadal de machos y hembras, la existencia de dimorfismo sexual y el periodo de vocalización de los machos. Los muestreos se efectuaron desde Septiembre de 2011 hasta Agosto de 2012. Dendropsophus sanborni exhibe un patrón reproductivo prolongado, dado que la actividad reproductiva ocurre durante gran parte del año. Se registraron machos en actividad de vocalización durante todos los meses excepto en julio. El ciclo espermatogénico puede ser caracterizado como potencialmente continuo. En los machos hubo correlación significativa entre la longitud hocico-cloaca y el volumen testicular medio. La especie presentó dimorfismo sexual en relación a la masa corporal, siendo las hembras mayores que los machos. Se encontraron hembras post-ovulatorias desde Marzo a Junio y una mayor proporción de hembras con ovocitos en diferentes estados de maduración, con abundantes ovocitos post-vitelogénicos, en los meses de Agosto a Diciembre.
Assuntos
Animais , Caracteres Sexuais , Gônadas , Reprodução/fisiologia , Vocalização AnimalRESUMO
Incidence and nematode population densities of plant-parasitic nematodes were determined in 64 samples of soil and grapevine roots collected from commercial vineyards in southern Spain between October 2003 and May 2005. In addition, a histopathological study was done of root-stock roots naturally infected by root-knot nematodes (Meloidogyne spp.). Nematodes infecting the rootstocks were identified according to conventional procedures, and the Meloidogyne spp. were furthermore identified by sequence characterized amplified region-polymerase chain reaction (SCAR-PCR) and isozyme esterase analyses. The most important plant-parasitic nematodes detected, in order of decreasing frequency of total soil infestation and root infection (percentage of samples), were Mesocriconema xenoplax (34.4%), Meloidogyne incognita (26.6%), Meloidogyne javanica (14.1%), Xiphinema index (12.5%), Xiphinema italiae (10.9%), Pratylenchus vulnus (6.3%), and Meloidogyne arenaria (1.6%). No disease symptoms were observed on aboveground plant parts of the infected grapevines, except for plants in some fields where soil was infested with the virus-vector nematodes X. index and X. italiae. Those grapevines showed a yellow mosaic pattern in leaves early in the growing season and the internode shortening characteristic of infections by Grapevine fanleaf virus. Rootstocks infected by root-knot nematodes (Meloidogyne spp.) showed distorted feeder roots and large- to moderate-sized root galls, present either singly or in clusters. Histopathology of galled roots showed a typical susceptible response to infection by root-knot nematodes: cellular alterations were induced in the cortex, endodermis, pericycle, and vascular system, including giant-cell formation and severe distortion of vascular tissues. Most Meloidogyne egg masses ocurred on the surface of the galled root tissues, a position that could facilitate dispersion of the nematode eggs and juveniles and the occurrence of secondary infections. Some of the grapevine rootstocks surveyed in this study (Paulsen 1103, Richter 110, Rupestris du Lot, and SO4) had previously been reported to be resistant to Meloidogyne spp.; however, the population densities of these nematodes found in soil and roots sampled in the present study, as well as the compatible host-parasite relationship revealed by histopathology, indicate a susceptible response to Meloidogyne spp. from southern Spain.