Microbial profiling of a suppressiveness-induced agricultural soil amended with composted almond shells lead to isolation of new biocontrol agents

Vida et al., 2016. Microbial profiling of a suppressiveness-induced agricultural soil amended with composted almond shells lead to isolation of new biocontrol agents. Biological and integrated control of plant pathogens IOBC-WPRS Bulletin Vol. 117, 2016 pp. 140-143This study focused on the microbial profile present in an agricultural soil that becomes suppressive after the application of composted almond shells (AS) as organic amendments. The role of microbes in the suppression of Rosellinia necatrix, the causative agent of avocado white root rot, was determined after heat-treatment and complementation experiments with different types of soil. Bacterial and fungal profiles based on the 16S rRNA gene and ITS sequencing, the soil under the influence of composted almond shells revealed an increase in Proteobacteria and Ascomycota groups, as well as a reduction in Acidobacteria and Xylariales (where R. necatrix is allocated). Complementary to these findings, functional analysis by GeoChip 4.6 confirmed the improvement of a group of specific probes included in the “soil benefit” category was present only in AS-amended soils, corresponding to specific microorganisms previously described as potential biocontrol agents, such as Pseudomonas spp., Burkholderia spp. or Actinobacteria. Based in such data, a model for the microbial-based suppressiveness is proposed and further isolation of representative microorganisms were performed.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Age and growth of the highly exploited narrownose smooth-hound (Mustelus schmitti) (Pisces: Elasmobranchii)

The narrownose smoothhound (Mustelus schmitti) is the most exploited elasmobranch of Argentina, Brazil, and Uruguay and is considered endangered (IUCN Red List of Threatened Species). Providing information on age and growth can improve efforts for conservation of this species. Therefore, our objective was to provide accurate estimates of the age structure and growth parameters for narrownose smooth-hound from Anegada Bay, an important shark nursery area in Argentina. In vertebrae of narrownose smooth-hound, we observed a pattern of alternating opaque and translucent bands and a yearly periodicity in the deposition of this pattern. Ages determined from vertebral band counts ranged from 0 to 11 years. Calculated longevity and total natural mortality rates were 20.87 years and 0.19/year for females and 12.24 years and 0.26/year for males, respectively. This species reached a size of approximately 400 mm in total length in the initial year of growth, and the age at first maturity was 7.61 years for females and 6.79 years for males. The slow growth and late age at maturity of the narrownose smoothhound indicate a need for additional conservation measures to rebuild the population and achieve a sustainable fishery in the 3 countries in which it is distributed.Fil: Molina, Juan Manuel. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Blasina, Gabriela Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto Argentino de Oceanografía. Universidad Nacional del Sur. Instituto Argentino de Oceanografía; ArgentinaFil: Lopez Cazorla, Andrea Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto Argentino de Oceanografía. Universidad Nacional del Sur. Instituto Argentino de Oceanografía; Argentin

Características histológicas del útero de Sympterygia acuta (Garman, 1877) y Sympterygia bonapartii (Müller & Henle, 1841) (Chondrichthyes; Rajidae)

Los condrictios constituyen un clado sumamente conservado y vulnerable a la explotación, que ha cobrado una importancia creciente en los últimos años. Sin embargo, existe poca información sobre los aspectos morfológicos relacionados con la reproducción; tanto así, que la estructura del útero en hembras grávidas sólo ha sido estudiada en profundidad en una especie ovípara. El presente trabajo describe las características morfológicas, histológicas e histoquímicas del útero de dos especies de Rájidos del género Sympterygia a lo largo del ciclo de vida. Ambas especies presentan úteros pares que desembocan independientemente en un seno urogenital común. El útero de las hembras inmaduras es filiforme, translúcido y poco especializado. Cuando se inicia el desarrollo sexual, todo el sistema genital exhibe un cambio direccional y progresivo. En todas las hembras estudiadas, independientemente del grado de madurez, se encontraron las mismas cuatro túnicas a lo largo del útero: una mucosa, compuesta por un epitelio de revestimiento y un corion, una submucosa, una muscular y una serosa. Los cambios mayores en la estructura del útero a lo largo del ciclo de vida ocurren a nivel de la mucosa. En el útero de hembras maduras, grávidas o no grávidas, se pueden reconocer tres regiones claramente diferenciables, craneal, media y caudal, que varían tanto en la estructura como en la afinidad histoquímica de la mucosa, así como en el grosor de la túnica muscular. Los resultados del presente trabajo se discuten en un contexto fisiológico, filogenético y adaptativo.Chondrichthyes constitute a conserve and vulnerable clade of increasing importance. However, there is scarce knowledge about the morphological aspects of the reproduction, so that only one work exists about the structure of the gravid uterus of an oviparous species. The work presented here describes the morphological, histological and histochemical remarks of the uterus of two Rajids of the Sympterygia genus along the life cycle. Both species presented paired uteri that lead independently to a common urogenital sinus. The uteri of immature females are filiform, translucent and unspecialized tubes. When sexual development starts, all the genital system displays a transitional and progressive change. In all studied females, independently of the maturing stage, the same four layers were present all over the uterus: a mucosa, which comprises the lining epithelium and lamina propria, a submucosa, a muscular and a serosa. The major changes in the structure of the uterus along the life cycle occur in the mucosa. The uteri of mature, gravid and nongravid, females presents three distinguishable areas, cranial, medial and caudal, that vary both in the structure and histochemistry of the mucosa and in the thickness of the muscular layer. The results presented here are discussed in a physiological, adaptive and phylogenetic context.Fil: Díaz Andrade, María Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Lopez Cazorla, Andrea Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Galíndez, Elena Juana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentin

Deciphering the suppressive soil microbiota from an avocado crop

Comunicación en pósterDifferent strategies based on ecological principles have been approached in sustainable agriculture causing positive effects, including the induction of soil suppressiveness against a wide range of soilborne pathogens. Suppressiveness against the phytopathogenic fungus Rosellinia necatrix was observed after the application of composted almond shells in avocado crops. Previous works have analyzed the use of this traditional strategy and applied new microbial community analysis techniques in order to help in the identification of targeted sustainable agricultural strategies. These studies have focused on the microbial profile from an induced-suppressive soil where the soil microbiome had a proven essential role. Microbial profiles based on the 16S rRNA gene and ITS regions sequencing were analysed and an increase in Gammaproteobacteria and Dothideomycetes groups, as well as a reduction in Xylariales (where R. necatrix is allocated) were observed. These results led to the bacterial isolation of different groups of Gammaproteobacteria from this suppressive soil in order to identify new strains with biological control properties. Different characterization tests were performed, and a final selection of representative strains belonging to the genus Pseudomonas and related groups showed, all of them, plant disease protection abilities. Moreover, using previously described biological control agents against R. necatrix, a bacterial synthetic community have been design in order to improve the knowledge of the multitrophic interactions that occur during biological control process.This work was supported by Plan Nacional I+D+I (MINECO, Spain) (AGL2014-52518-C2-IR) and co-financed by FEDER funds (EU). C.Vida was supported by a PhD fellowship from the FPI program of MINECO. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Isolation, characterization and selection of Pseudomonas spp. as biological control agents from a suppressive soil

Bacterial profile from a suppressive soil against Rosellinia necatrix, fungal pathogen of avocado roots, were obtained by 16S rRNA gene sequencing. The results revealed a significant increase in the bacterial class of Gammaproteobacteria, especially in some antagonistic representatives of Pseudomonas spp. For this reason, a collection of 246 bacterial isolates was obtained from this suppressive soil in order to identify new strains with antifungal activity against this fungal phytopathogen. First, we performed an isolation on a selective medium for Pseudomonas-like microorganisms. Then, we used different characterization tests in order to analyse the bacterial collection, including the identification of the general metabolic profile of glucose, the profiling of antifungals produced, both the putative production of antifungal compounds and lytic exoenzymes, as well as the evaluation of traits related with beneficial effects on plants. A final selection of representative strains resulted in antifungal isolates belonging to the genus Pseudomonas and related groups. 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.This work was supported by Plan Nacional I+D+I from MINECO (Spain) (AGL2014-52518-C2-IR) and co-financed by FEDER funds (EU). C.Vida was supported by a PhD fellowship from the FPI program of MINECO.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Foliculogénesis Ovárica en la Raya Marmorada, Sympterygia bonapartii (Müller & Henle, 1841) (Chondrichthyes, Rajidae)

Los Condrictios se han convertido en un recurso económico importante en las últimas décadas, siendo Argentina uno de los países que más explota tiburones y rayas, incluso a niveles que exceden los límites de varias especies. A pesar de esto, es poco lo que se conoce sobre la biología reproductiva de este grupo, particularmente en especies del Hemisferio Sur. En este trabajo se estudian los estadios más relevantes de la foliculogénesis en Sympterygia bonapartii. Los resultados muestran que las ovogonias están presentes tanto en ejemplares inmaduros como subadultos. Las características más importantes que varían a lo largo del desarrollo folicular son el número de capas y tipos celulares que constituyen el epitelio folicular, el desarrollo de proyecciones de las células de la granulosa y el grado de desarrollo de las tecas. Las células foliculares son, al menos, de dos tipos y ambos emiten proyecciones que atraviesan la zona pelúcida. La teca externa presenta características compatibles con la actividad sintética. Folículos atrésicos de distintos tamaños están presentes en ejemplares de todos los estadios de madurez sexual. Estos resultados se discuten en un marco fisiológico y adaptativo.Chondrichthyes have become an important economic resource in the last decades, with Argentina as one of the countries that exploits more sharks and skates, even at levels that exceed de limits of many species. However, there is scarce knowledge of the reproductive biology of this group, particularly from species inhabiting the Southern hemisphere. This work shows the most relevant facts during folliculogenesis in Sympterygia bonapartii. Results show that germinal cells are present in immature and maturing females. The most important facts that vary along de follicular development are the number of types and layers of follicular cells, the establishment of thin projections from the follicular cells and the degree of development of the thecae. Follicular cells are, at least, of two different types and both of them emit projections that break through the zona pellucida. The outer theca shows signs of synthetic activity. Atretic follicles of different sizes are present in exemplars of all the reproductive stages. These results are discussed in a physiological and adaptive context.Fil: Díaz Andrade, María Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Galíndez, Elena Juana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Lopez Cazorla, Andrea Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Estecondo, Silvia. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentin

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

Análisis de la comunidad microbiana de un suelo supresivo en el cultivo del aguacate

Comunicación oralLa podredumbre blanca de la raíz causada por el hongo fitopatógeno Rosellinia necatrix, es uno de los problemas más graves del cultivo del aguacate en el área mediterránea. Desde hace años, el manejo integrado de la enfermedad ha incluido la aplicación de enmiendas orgánicas como estrategia para mejorar el estado fitosanitario de los suelos agrícolas. En trabajos anteriores, se demostró el efecto positivo que tenía la aplicación de cáscara de almendra compostada en el control biológico del patógeno, relacionado con los cambios inducidos en las propiedades fisicoquímicas y microbianas del suelo. En este trabajo, nos centramos en analizar el papel de la comunidad microbiana de estos suelos denominados suelos supresivos. Para ello, se llevaron a cabo ensayos “in vitro” frente a R. necatrix que mostraron que la adición de cáscara de almendra compostada al suelo, causa un aumento de la supresividad frente a R. necatrix asociada a la microbiota. El uso de técnicas de genómica molecular nos ha permitido conocer la composición y el potencial funcional (GeoChip®) de la microbiota de estos suelos. Concretamente, esta actividad supresiva era llevada a cabo por algunos grupos específicos de microorganismos que realizaban una actividad integrada. El aislamiento y caracterización de bacterias cultivables de la clase Gammaproteobacteria, mostró la capacidad de estos aislados para controlar el índice de enfermedad causado por el patógeno, utilizando diferentes estrategias de control biológico como la posible producción de compuestos antifúngicos, exoenzimas líticas o la promoción de crecimiento vegetal. Además, se ha diseñado un consorcio bacteriano artificial formado por representantes de esta clase de bacterias de suelo, descritos en trabajos anteriores como agentes de control biológico frente a R. necatrix, con el fin de profundizar en el conocimiento de los posibles mecanismos implicados en la supresividad, así como, en el patrón de colonización de raíz de este consorcio bacteriano artificial.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech