Artículos de revista DPA - NES Aldizkari artikuluak

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Open Access
Adiposity and adipogenic gene expression in four different muscles in beef cattle
(Public Library of Science, 2017) Martínez del Pino, Lara; Arana Navarro, Ana; Alfonso Ruiz, Leopoldo; Mendizábal Aizpuru, José Antonio; Soret Lafraya, Beatriz; Producción Agraria; Nekazaritza Ekoizpena
Anatomical site and divergent functionalities of muscles can be related to differences in IMF content, metabolism and adipogenic gene expression. Then, potential differences in different muscles in beef cattle were studied. As a second objective, the main sources of experimental variability associated to RT-qPCR results were analyzed following a nested design in order to implement appropriate experimental designs minimizing gene expression variability. To perform the study Longissimus thoracis (LT), Semitendinosus (SM), Masseter (MS), Sternomandibularis (ST) and subcutaneous adipose tissue (SAT) samples of Pirenaica young bulls (n = 4) were collected for IMF, collagen and protein quantification, analysis of adipocyte size distribution and gene expression (PPARG, CEBPA, FAPB4 and WNT10B). A greater IMF content was observed in MS and SM muscles, which had a bimodal adipocyte size distribution while it was unimodal in the muscles LT and ST. This suggest that the different IMF accretion in the muscles studied might be related to different rates of hyperplasia and hypertrophy and that IMF might develop later in LT and ST muscles. The former differences were not mirrored by the expression of the genes analyzed, which might be related to the different contribution of mature and non-mature adipocytes to the total gene expression. When comparing IMF and SAT gene expression, late and early developing tissues respectively, expression of PPARG, CEBPA and FABP4 was higher in the SAT, in agreement with bigger cell size and numbers. The variability study indicates that the analytical factors that add higher variability to the gene expression are the sampling and RT and therefore, it would be appropriate to include those replicates in the design of future experiments. Based on the results, the use of MS and SM muscles could allow less expensive experimental designs and bigger sample size that could permit the detection of lower relevant differences in gene expression.
Open Access
Análisis de la relación entre el espesor de la grasa dorsal y el tamaño de los adipocitos en cerdas reproductoras
(Asociación Interprofesional para el Desarrollo Agrario, 2007) Mendizábal Aizpuru, José Antonio; Abadía Durán, Silvia; Abaurrea Aramburu, Eneko; Alfonso Ruiz, Leopoldo; Producción Agraria; Nekazaritza Ekoizpena
La correcta gestión de las reservas grasas durante el ciclo productivo de las cerdas reproductoras es un factor clave para la obtención de unos buenos resultados productivos. El principal depósito graso en las cerdas lo constituye el tejido graso subcutáneo, por lo que la medición del espesor graso dorsal es un buen indicador del estado de las reservas corporales. Aunque existen diversos puntos para la medición del espesor, se recomienda hacerlo a nivel de la última costilla flotante, punto que se conoce como P2 (Quiniou, 2004). En esta localización se pueden distinguir hasta tres capas distintas de grasa subcutánea (Moody y Zobrisky, 1966). Distintos trabajos han observado un desarrollo diferencial de las tres capas durante el crecimiento de los animales (Alfonso, 2004; Fortin, 1986), así como diferencias en su actividad lipogénica (Cámara et al., 1996). Por tanto, el estudio de las características de cada capa considerada como distinto depósito graso puede ser importante para conocer los mecanismos de acumulación y movilización de las reservas grasas. En este sentido, en el presente trabajo se estudia la relación entre el espesor de la grasa y el tamaño de los adipocitos de cada una de las capas descritas en la grasa subcutánea dorsal.
Open Access
Analysis of the genetic diversity and structure across a wide range of germplasm reveals prominent gene flow in apple at the European level
(BioMed Central, 2016) Urrestarazu Vidart, Jorge; Denancé, Caroline; Ravon, Elisa; Guyader, Arnaud; Guisnel, Rémi; Feugey, Laurence; Poncet, Charles; Lateur, Marc; Houben, Patrick; Ordidge, Matthew; Fernández Fernández, Felicidad; Evans, Kate M.; Paprstein, Frantisek; Sedlak, Jiri; Nybom, Hilde; Garkava Gustavsson, Larisa; Miranda Jiménez, Carlos; Gassmann, Jennifer; Kellerhals, Markus; Suprun, Ivan; Pikunova, Anna V.; Krasova, Nina G.; Torutaeva, Elnura; Dondini, Luca; Tartarini, Stefano; Laurens, François; Durel, Charles Eric; Producción Agraria; Nekazaritza Ekoizpena
Background: The amount and structure of genetic diversity in dessert apple germplasm conserved at a European level is mostly unknown, since all diversity studies conducted in Europe until now have been performed on regional or national collections. Here, we applied a common set of 16 SSR markers to genotype more than 2,400 accessions across 14 collections representing three broad European geographic regions (North + East, West and South) with the aim to analyze the extent, distribution and structure of variation in the apple genetic resources in Europe. Results: A Bayesian model-based clustering approach showed that diversity was organized in three groups, although these were only moderately differentiated (FST = 0.031). A nested Bayesian clustering approach allowed identification of subgroups which revealed internal patterns of substructure within the groups, allowing a finer delineation of the variation into eight subgroups (FST = 0.044). The first level of stratification revealed an asymmetric division of the germplasm among the three groups, and a clear association was found with the geographical regions of origin of the cultivars. The substructure revealed clear partitioning of genetic groups among countries, but also interesting associations between subgroups and breeding purposes of recent cultivars or particular usage such as cider production. Additional parentage analyses allowed us to identify both putative parents of more than 40 old and/or local cultivars giving interesting insights in the pedigree of some emblematic cultivars. Conclusions: The variation found at group and subgroup levels may reflect a combination of historical processes of migration/selection and adaptive factors to diverse agricultural environments that, together with genetic drift, have resulted in extensive genetic variation but limited population structure. The European dessert apple germplasm represents an important source of genetic diversity with a strong historical and patrimonial value. The present work thus constitutes a decisive step in the field of conservation genetics. Moreover, the obtained data can be used for defining a European apple core collection useful for further identification of genomic regions associated with commercially important horticultural traits in apple through genome-wide association studies.
Open Access
Aportación veterinaria en los inicios de la avicultura española
(Consejo General de Veterinarios de España, 2015) Mendizábal Aizpuru, José Antonio; Producción Agraria; Nekazaritza Ekoizpena
El autor pretende mostrar la aportación de eminentes veterinarios en el desarrollo de la avicultura española, desde mediados del siglo XIX hasta la Guerra Civil de 1936.
Open Access
Assessment of the genetic and phenotypic diversity maintained in apple core collections constructed by using either agro-morphologic or molecular marker data
(Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), 2009) Santesteban García, Gonzaga; Miranda Jiménez, Carlos; Royo Díaz, José Bernardo; Producción Agraria; Nekazaritza Ekoizpena
Las colecciones nucleares de plantas pueden formarse a partir de la información obtenida en caracterizaciones morfológicas, agronómicas o eco-geográficas, así como a partir de caracterizaciones bioquímicas o moleculares. Un aspecto poco estudiado de la formación y validación de colecciones nucleares es la capacidad de éstas para conservar la estructura y diversidad de la colección global en aquellos caracteres que no han sido empleados para formar la colección nuclear. En el presente estudio se determinaron tres colecciones nucleares para el Banco de Germoplasma de Manzano de la Universidad Pública de Navarra: para el primero se emplearon los datos de caracterización de 10 marcadores SSR, la segunda fue formada a partir de la caracterización hecha con 12 loci de isoenzimas, y la tercera se construyó a partir de la información proporcionada por 23 caracteres morfo-agronómicos. Se comparó la estructura y diversidad genotípica y fenotípica de las tres colecciones, entre sí y respecto de la colección original, para determinar el impacto que sobre dicha estructura y diversidad tenía el tipo de dato con que se formó la colección nuclear. Las tres colecciones conservaron una diversidad similar y no difirieron de la original en sus índices de Nei y Shannon-Weaver, y siempre se mantuvieron las frecuencias de las clases alélicas o fenotípicas. En conjunto, el tipo de caracterización empleada en la formación de la colección nuclear tuvo escasa influencia sobre la diversidad retenida, de lo que se concluye que en el manzano los microsatélites son especialmente adecuados para formar dichas colecciones.
Open Access
B regulates IS256-mediated Staphylococcus aureus biofilm phenotypic variation
(American Society for Microbiology, 2007) Valle Turrillas, Jaione; Vergara Irigaray, Marta; Merino Barberá, Nekane; Penadés, José R.; Lasa Uzcudun, Íñigo; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Biofilm formation in Staphylococcus aureus is subject to phase variation, and biofilm-negative derivatives emerge sporadically from a biofilm-positive bacterial population. To date, the only known mechanism for generating biofilm phenotypic variation in staphylococci is the reversible insertion/excision of IS256 in biofilm-essential genes. In this study, we present evidence suggesting that the absence of the σB transcription factor dramatically increases the rate of switching to the biofilm-negative phenotype in the clinical isolate S. aureus 15981, under both steady-state and flow conditions. The phenotypic switching correlates with a dramatic increase in the number of IS256 copies in the chromosomes of biofilm-negative variants, as well as with an augmented IS256 insertion frequency into the icaC and the sarA genes. IS256-mediated biofilm switching is reversible, and biofilm-positive variants could emerge from biofilm-negative σB mutants. Analysis of the chromosomal insertion frequency using a recombinant IS256 element tagged with an erythromycin marker showed an almost three-times-higher transposition frequency in a ΔσB strain. However, regulation of IS256 activity by σB appears to be indirect, since transposase transcription is not affected in the absence of σB and IS256 activity is inhibited to wild-type levels in a ΔσB strain under NaCl stress. Overall, our results identify a new role for σB as a negative regulator of insertion sequence transposition and support the idea that deregulation of IS256 activity abrogates biofilm formation capacity in S. aureus.
Open Access
Bacillus thuringiensis toxins: an overview of their biocidal activity
(MDPI, 2014) Palma Dovis, Leopoldo; Muñoz Labiano, Delia; Berry, Colin; Murillo Martínez, Jesús; Caballero Murillo, Primitivo; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Bacillus thuringiensis (Bt) is a Gram positive, spore-forming bacterium that synthesizes parasporal crystalline inclusions containing Cry and Cyt proteins, some of which are toxic against a wide range of insect orders, nematodes and human-cancer cells. These toxins have been successfully used as bioinsecticides against caterpillars, beetles, and flies, including mosquitoes and blackflies. Bt also synthesizes insecticidal proteins during the vegetative growth phase, which are subsequently secreted into the growth medium. These proteins are commonly known as vegetative insecticidal proteins (Vips) and hold insecticidal activity against lepidopteran, coleopteran and some homopteran pests. A less well characterized secretory protein with no amino acid similarity to Vip proteins has shown insecticidal activity against coleopteran pests and is termed Sip (secreted insecticidal protein). Bin-like and ETX_MTX2-family proteins (Pfam PF03318), which share amino acid similarities with mosquitocidal binary (Bin) and Mtx2 toxins, respectively, from Lysinibacillus sphaericus, are also produced by some Bt strains. In addition, vast numbers of Bt isolates naturally present in the soil and the phylloplane also synthesize crystal proteins whose biological activity is still unknown. In this review, we provide an updated overview of the known active Bt toxins to date and discuss their activities.
Open Access
Bap, a Staphylococcus aureus surface protein involved in biofilm formation
(American Society for Microbiology, 2001) Cucarella, Carme; Solano Goñi, Cristina; Valle Turrillas, Jaione; Amorena Zabalza, Beatriz; Lasa Uzcudun, Íñigo; Penadés, José R.; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Gobierno de Navarra / Nafarroako Gobernua
Identification of new genes involved in biofilm formation is needed to understand the molecular basis of strain variation and the pathogenic mechanisms implicated in chronic staphylococcal infections. A biofilm-producing Staphylococcus aureus isolate was used to generate biofilm-negative transposon (Tn917) insertion mutants. Two mutants were found with a significant decrease in attachment to inert surfaces (early adherence), intercellular adhesion, and biofilm formation. The transposon was inserted at the same locus in both mutants. This locus (bap [for biofilm associated protein]) encodes a novel cell wall associated protein of 2,276 amino acids (Bap), which shows global organizational similarities to surface proteins of gram-negative (Pseudomonas aeruginosa andSalmonella enterica serovar Typhi) and gram-positive (Enteroccocus faecalis) microorganisms. Bap's core region represents 52% of the protein and consists of 13 successive nearly identical repeats, each containing 86 amino acids. bap was present in a small fraction of bovine mastitis isolates (5% of the 350S. aureus isolates tested), but it was absent from the 75 clinical human S. aureus isolates analyzed. All staphylococcal isolates harboring bap were highly adherent and strong biofilm producers. In a mouse infection modelbap was involved in pathogenesis, causing a persistent infection.
Open Access
Calidad de la canal y de la carne caballar de raza Burguete
(Asociación Interprofesional para el Desarrollo Agrario, 2001) Sarriés Martínez, María Victoria; Larrea Reta, Izaskun; Induráin Báñez, Gregorio; Goñi Turumbay, Virginia; Eguinoa Ancho, Paola; Gorraiz, C.; Martín, M.; Alzueta Aldunate, María Jesús; Beriain Apesteguía, María José; Pérez de Muniáin, A.; Producción Agraria; Nekazaritza Ekoizpena
La raza caballar Burguete es una raza autóctona de Navarra que en el pasado disfrutó de gran prestigio como animal de trabajo en la Comunidad Foral y zonas limítrofes. Hoy en día por su clara aptitud cárnica es importante mantener dicho patrimonio genético y mejorar los productos obtenidos para la promoción de esta raza estableciendo criterios de tipificación (Pérez de Muniáin et al, 2000). Se trata de un producto poco conocido en el mercado de la carne, aunque en la actualidad está despertando interés debido a sus reconocidas cualidades nutritivas (producto natural y sano) como consecuencia del bajo contenido en grasa. El objetivo del presente trabajo ha sido la caracterización de la canal y de la carne caballar de raza Burguete.
Open Access
Caracterización de la distribución del tamaño de los adipocitos para el estudio del tejido adiposo en producción animal
(Asociación Interprofesional para el Desarrollo Agrario, 2016) Alfonso Ruiz, Leopoldo; Mendizábal Aizpuru, José Antonio; Producción Agraria; Nekazaritza Ekoizpena
El tamaño de los adipocitos, principales células constituyentes del tejido adiposo, ha sido objeto de numerosos estudios por el hecho de estar relacionado con el grado de desarrollo y la actividad metabólica de ese tejido. Su análisis resulta en ocasiones complejo dada la bimodalidad de su distribución. Este trabajo presenta un método de análisis basado en contrastar, en primer lugar la unimodalidad de la distribución del tamaño de los adipocitos, frente a una distribución bimodal. Posteriormente propone algunos parámetros para describir adecuadamente la distribución bimodal. Para su aplicación se desarrolló un sencillo programa informático que se utiliza, a modo de ejemplo, para analizar los datos de dos trabajos, previamente publicados, sobre el desarrollo adipocitario. Los resultados muestran el interés de utilizar contrastes de bimodalidad frente a la mera inspección visual de los histogramas de distribución. Además, cuando la hipótesis de unimodalidad es rechazada, la utilización de parámetros descriptivos de bimodalidad, como el porcentaje de adipocitos por encima del punto de inflexión entre ambas modas, permite una comparación más adecuada entre tratamientos experimentales.
Open Access
Changes in race-specific virulence in Pseudomonas syringae pv. phaseolicola are associated with a chimeric transposable element and rare deletion events in a plasmid-borne pathogenicity island
(American Society for Microbiology, 2005) Rivas Mena, Luis; Mansfield, John W.; Tsiamis, George; Jackson, Robert W.; Murillo Martínez, Jesús; Producción Agraria; Nekazaritza Ekoizpena
Virulence for bean and soybean is determined by effector genes in a plasmid-borne pathogenicity island (PAI) in race 7 strain 1449B of Pseudomonas syringae pv. phaseolicola. One of the effector genes, avrPphF, confers either pathogenicity, virulence, or avirulence depending on the plant host and is absent from races 2, 3, 4, 6, and 8 of this pathogen. Analysis of cosmid clones and comparison of DNA sequences showed that the absence of avrPphF from strain 1448A is due to deletion of a continuous 9.5-kb fragment. The remainder of the PAI is well conserved in strains 1448A and 1449B. The left junction of the deleted region consists of a chimeric transposable element generated from the fusion of homologs of IS1492 from Pseudomonas putida and IS1090 from Ralstonia eutropha. The borders of the deletion were conserved in 66 P. syringae pv. phaseolicola strains isolated in different countries and representing the five races lacking avrPphF. However, six strains isolated in Spain had a 10.5-kb deletion that extended 1 kb further from the right junction. The perfect conservation of the 28-nucleotide right repeat of the IS1090 homolog in the two deletion types and in the other 47 insertions of the IS1090 homolog in the 1448A genome strongly suggests that the avrPphF deletions were mediated by the activity of the chimeric mobile element. Our data strongly support a clonal origin for the races of P. syringae pv. phaseolicola lacking avrPphF.
Open Access
Characterisation of the mgo operon in Pseudomonas syringae pv. syringae UMAF0158 that is required for mangotoxin production
(BioMed Central, 2012) Arrebola, Eva; Carrión, Víctor J.; Cazorla, Francisco M.; Pérez García, Alejandro; Murillo Martínez, Jesús; Vicente, Antonio de; Producción Agraria; Nekazaritza Ekoizpena
Background: Mangotoxin is an antimetabolite toxin that is produced by strains of Pseudomonas syringae pv. syringae; mangotoxin-producing strains are primarily isolated from mango tissues with symptoms of bacterial apical necrosis. The toxin is an oligopeptide that inhibits ornithine N-acetyl transferase (OAT), a key enzyme in the biosynthetic pathway of the essential amino acids ornithine and arginine. The involvement of a putative nonribosomal peptide synthetase gene (mgoA) in mangotoxin production and virulence has been reported. Results: In the present study, we performed a RT-PCR analysis, insertional inactivation mutagenesis, a promoter expression analysis and terminator localisation to study the gene cluster containing the mgoA gene. Additionally, we evaluated the importance of mgoC, mgoA and mgoD in mangotoxin production. A sequence analysis revealed an operon-like organisation. A promoter sequence was located upstream of the mgoB gene and was found to drive lacZ transcription. Two terminators were located downstream of the mgoD gene. RT-PCR experiments indicated that the four genes (mgoBCAD) constitute a transcriptional unit. This operon is similar in genetic organisation to those in the three other P. syringae pathovars for which complete genomes are available (P. syringae pv. syringae B728a, P. syringae pv. tomato DC3000 and P. syringae pv. phaseolicola 1448A). Interestingly, none of these three reference strains is capable of producing mangotoxin. Additionally, extract complementation resulted in a recovery of mangotoxin production when the defective mutant was complemented with wild-type extracts. Conclusions: The results of this study confirm that mgoB, mgoC, mgoA and mgoD function as a transcriptional unit and operon. While this operon is composed of four genes, only the last three are directly involved in mangotoxin production.
Open Access
Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV): natural occurrence and efficacy as a biological insecticide on young banana plants in greenhouse and open-field conditions on the Canary Islands
(Public Library of Science, 2017) Fuentes Barrera, Ernesto Gabriel; Hernández Suárez, Estrella; Simón de Goñi, Oihane; Williams, Trevor; Caballero Murillo, Primitivo; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Chrysodeixis chalcites, an important pest of banana crops on the Canary Islands, is usually controlled by chemical insecticides. The present study aimed to evaluate the efficacy of the most prevalent isolate of the Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV, Baculoviridae) as a biological insecticide. Overall the prevalence of ChchNPV infection in C. chalcites populations was 2.3% (103 infected larvae out of 4,438 sampled), but varied from 0±4.8% on Tenerife and was usually low (0±2%) on the other islands. On Tenerife, infected larvae were present at 11 out of 17 plantations sampled. The prevalence of infection in larvae on bananas grown under greenhouse structures was significantly higher (3%) than in open-field sites (1.4%). The ChchNPV-TF1 isolate was the most abundant and widespread of four genetic variants of the virus. Application of 1.0x109 viral occlusion bodies (OBs)/l of ChchNPV-TF1 significantly reduced C. chalcites foliar damage in young banana plants as did commonly used pesticides, both in greenhouse and open-field sites. The insecticidal efficacy of ChchNPV-TF1 was similar to that of indoxacarb and a Bacillus thuringiensis (Bt)- based insecticide in one year of trials and similar to Bt in the following year of trails in greenhouse and field crops. However, larvae collected at different time intervals following virus treatments and reared in the laboratory experienced 2±7 fold more mortality than insects from conventional insecticide treatments. This suggests that the acquisition of lethal dose occurred over an extended period (up to 7 days) compared to a brief peak in larvae on plants treated with conventional insecticides. These results should prove useful for the registration of a ChchNPV-based insecticide for integrated management of this pest in banana crops on the Canary Islands.
Open Access
A Chrysodeixis chalcites single-nucleocapsid nucleopolyhedrovirus population from the Canary Islands is genotypically structured to maximize survival
(American Society for Microbiology, 2013) Bernal Rodríguez, Alexandra; Simón de Goñi, Oihane; Williams, Trevor; Muñoz Labiano, Delia; Caballero Murillo, Primitivo; Producción Agraria; Nekazaritza Ekoizpena; Gobierno de Navarra / Nafarroako Gobernua, IIQ14065:RI1
A Chrysodeixis chalcites single-nucleocapsid nucleopolyhedrovirus wild-type isolate from the Canary Islands, Spain, named ChchSNPV-TF1 (ChchTF1-wt), appears to have great potential as the basis for a biological insecticide for control of the pest. An improved understanding of the genotypic structure of this wild-type strain population should facilitate the selection of genotypes for inclusion in a bioinsecticidal product. Eight genetically distinct genotypes were cloned in vitro: ChchTF1-A to ChchTF1-H. Quantitative real-time PCR (qPCR) analysis confirmed that ChchTF1-A accounted for 36% of the genotypes in the wild-type population. In bioassays, ChchTF1-wt occlusion bodies (OBs) were significantly more pathogenic than any of the component single-genotype OBs, indicating that genotype interactions were likely responsible for the pathogenicity phenotype of wild-type OBs. However, the wild-type population was slower killing and produced higher OB yields than any of the single genotypes alone. These results strongly suggested that the ChchTF1-wt population is structured to maximize its transmission efficiency. Experimental OB mixtures and cooccluded genotype mixtures containing the most abundant and the rarest genotypes, at frequencies similar to those at which they were isolated, revealed a mutualistic interaction that restored the pathogenicity of OBs. In OB and cooccluded mixtures containing only the most abundant genotypes, ChchTF1-ABC, OB pathogenicity was even greater than that of wild-type OBs. The ChchTF1-ABC cooccluded mixture killed larvae 33 h faster than the wild-type population and remained genotypically and biologically stable throughout five successive passages in vivo. In conclusion, the ChchTF1- ABC mixture shows great potential as the active ingredient of a bioinsecticide to control C. chalcites in the Canary Islands.
Open Access
Co-infection with iflaviruses influences the insecticidal properties of Spodoptera exigua multiple nucleopolyhedrovirus occlusion bodies: implications for the production and biosecurity of baculovirus insecticides
(Public Library of Science, 2017) Carballo Palos, Arkaitz; Murillo Pérez, Rosa; Jakubowska, Agata; Herrero, Salvador; Williams, Trevor; Caballero Murillo, Primitivo; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Biological insecticides based on Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) can efficiently control S. exigua larvae on field and greenhouse crops in many parts of the world. Spanish wild populations and laboratory colonies of S. exigua are infected by two iflaviruses (SeIV-1 and SeIV-2). Here we evaluated the effect of iflavirus co-infection on the insecticidal characteristics of SeMNPV occlusion bodies (OBs). Overall, iflavirus coinoculation consistently reduced median lethal concentrations (LC50) for SeMNPV OBs compared to larvae infected with SeMNPV alone. However, the speed of kill of SeMNPV was similar in the presence or absence of the iflaviruses. A reduction of the weight gain (27%) associated with iflavirus infection resulted in a 30% reduction in total OB production per larva. Adult survivors of SeMNPV OB inoculation were examined for covert infection. SeMNPV DNA was found to be present at a high prevalence in all SeIV-1 and SeIV-2 coinfection treatments. Interestingly, co-inoculation of SeMNPV with SeIV-2 alone or in mixtures with SeIV-1 resulted in a significant increase in the SeMNPV load of sublethally infected adults, suggesting a role for SeIV-2 in vertical transmission or reactivation of sublethal SeMNPV infections. In conclusion, iflaviruses are not desirable in insect colonies used for large scale baculovirus production, as they may result in diminished larval growth, reduced OB production and, depending on their host-range, potential risks to non-target Lepidoptera.
Open Access
Comparative and transcriptional analysis of the predicted secretome in the lignocellulose-degrading basidiomycete fungus Pleurotus ostreatus
(Wiley, 2016) Alfaro Sánchez, Manuel; Castanera Andrés, Raúl; Lavín Trueba, José Luis; Oguiza Tomé, José Antonio; Ramírez Nasto, Lucía; Pisabarro de Lucas, Gerardo; Producción Agraria; Nekazaritza Ekoizpena; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Fungi interact with their environment by secreting proteins to obtain nutrients, elicit responses and modify their surroundings. Because the set of proteins secreted by a fungus is related to its lifestyle, it should be possible to use it as a tool to predict fungal lifestyle. To test this hypothesis, we bioinformatically identified 538 and 554 secretable proteins in the monokaryotic strains PC9 and PC15 of the white rot basidiomycete Pleurotus ostreatus. Functional annotation revealed unknown functions (37.2%), glycosyl hydrolases (26.5%) and redox enzymes (11.5%) as the main groups in the two strains. When these results were combined with RNA‐seq analyses, we found that the relative importance of each group was different in different strains and culture conditions and the relevance of the unknown function proteins was enhanced. Only a few genes were actively expressed in a given culture condition in expanded multigene families, suggesting that family expansi on could increase adaptive opportunities rather than activity under a specific culture condition. Finally, we used the set of P. ostreatus secreted proteins as a query to search their counterparts in other fungal genomes and found that the secretome profiles cluster the tested basidiomycetes into lifestyle rather than phylogenetic groups.
Open Access
Comparative description of growth, fat deposition, carcass and meat quality characteristics of Basque and Large White pigs
(EDP Sciences, 2005) Alfonso Ruiz, Leopoldo; Mourot, Jacques; Insausti Barrenetxea, Kizkitza; Mendizábal Aizpuru, José Antonio; Arana Navarro, Ana; Producción Agraria; Nekazaritza Ekoizpena; Gobierno de Navarra / Nafarroako Gobernua
Characteristics of growth, fat deposition, carcass and meat quality of pigs from the Basque Black Pied breed were described and compared with those of Large White pigs. Four pens, two per breed, of eleven pigs born during the same two week period, were simultaneously fattened and slaughtered, under the same conditions. The experiment was carried out over a fixed duration (124 days) and slaughter was carried out at a fixed average age (202 days). Basque pigs showed lower growth and feed efficiency and higher backfat depth (2.6 vs. 1.7 cm, P < 0.001) than Large White pigs. The difference was especially noticeable in the middle subcutaneous fat layer (0.5 cm, P < 0.001). The meat of Basque pigs was darker, redder, more marbled, and with higher pH values than in Large White pigs. Differences in fatty acid composition were observed between breeds but they were not statistically significant (P > 0.05) because of high variability observed between animals. The Basque breed exhibited an early and higher adipose development and a higher activity of enzymes responsible for lipid synthesis than the Large White. The diameter of intramuscular adipose cells was larger in Basque (40.2 vs. 33.0 μm, P < 0.001) than in Large White pigs. The results show the particular characteristics of the Basque breed as compared to pig lines highly selected for lean growth efficiency.
Open Access
Comparative genomic analysis of two-component regulatory proteins in pseudomonas syringae
(BioMed Central, 2007) Lavín Trueba, José Luis; Kiil, Kristoffer; Resano Aranibar, Ohiana; Ussery, David W.; Oguiza Tomé, José Antonio; Producción Agraria; Nekazaritza Ekoizpena
Background: Pseudomonas syringae is a widespread bacterial plant pathogen, and strains of P. syringae may be assigned to different pathovars based on host specificity among different plant species. The genomes of P. syringae pv. syringae (Psy) B728a, pv. tomato (Pto) DC3000 and pv. phaseolicola (Pph) 1448A have been recently sequenced providing a major resource for comparative genomic analysis. A mechanism commonly found in bacteria for signal transduction is the two-component system (TCS), which typically consists of a sensor histidine kinase (HK) and a response regulator (RR). P. syringae requires a complex array of TCS proteins to cope with diverse plant hosts, host responses, and environmental conditions. Results: Based on the genomic data, pattern searches with Hidden Markov Model (HMM) profiles have been used to identify putative HKs and RRs. The genomes of Psy B728a, Pto DC3000 and Pph 1448A were found to contain a large number of genes encoding TCS proteins, and a core of complete TCS proteins were shared between these genomes: 30 putative TCS clusters, 11 orphan HKs, 33 orphan RRs, and 16 hybrid HKs. A close analysis of the distribution of genes encoding TCS proteins revealed important differences in TCS proteins among the three P. syringae pathovars. Conclusion: In this article we present a thorough analysis of the identification and distribution of TCS proteins among the sequenced genomes of P. syringae. We have identified differences in TCS proteins among the three P. syringae pathovars that may contribute to their diverse host ranges and association with plant hosts. The identification and analysis of the repertoire of TCS proteins in the genomes of P. syringae pathovars constitute a basis for future functional genomic studies of the signal transduction pathways in this important bacterial phytopathogen.
Open Access
Comparative genomics of Ceriporiopsis subvermispora and Phanerochaete chrysosporium provide insight into selective ligninolysis
(National Academy of Sciences, 2012) Fernández Fueyo, Elena; Ruiz Dueñas, Francisco J.; Ferreira, Patricia; Floudas, Dimitrios; Lavín Trueba, José Luis; Oguiza Tomé, José Antonio; Pérez Garrido, María Gumersinda; Pisabarro de Lucas, Gerardo; Ramírez Nasto, Lucía; Santoyo Santos, Francisco; Producción Agraria; Nekazaritza Ekoizpena
Efficient lignin depolymerization is unique to the wood decay basidiomycetes, collectively referred to as white rot fungi. Phanerochaete chrysosporium simultaneously degrades lignin and cellulose, whereas the closely related species, Ceriporiopsis subvermispora, also depolymerizes lignin but may do so with relatively little cellulose degradation. To investigate the basis for selective ligninolysis, we conducted comparative genome analysis of C. subvermispora and P. chrysosporium. Genes encoding manganese peroxidase numbered 13 and five in C. subvermispora and P. chrysosporium, respectively. In addition, the C. subvermispora genome contains at least seven genes predicted to encode laccases, whereas the P. chrysosporium genome contains none. We also observed expansion of the number of C. subvermispora desaturase-encoding genes putatively involved in lipid metabolism. Microarray-based transcriptome analysis showed substantial up-regulation of several desaturase and MnP genes in wood-containing medium. MS identified MnP proteins in C. subvermispora culture filtrates, but none in P. chrysosporium cultures. These results support the importance of MnP and a lignin degradation mechanism whereby cleavage of the dominant nonphenolic structures is mediated by lipid peroxidation products. Two C. subvermispora genes were predicted to encode peroxidases structurally similar to P. chrysosporium lignin peroxidase and, following heterologous expression in Escherichia coli, the enzymes were shown to oxidize high redox potential substrates, but not Mn2+. Apart from oxidative lignin degradation, we also examined cellulolytic and hemicellulolytic systems in both fungi. In summary, the C. subvermispora genetic inventory and expression patterns exhibit increased oxidoreductase potential and diminished cellulolytic capability relative to P. chrysosporium.
Open Access
Comparative genomics of Coniophora olivacea reveals different patterns of genome expansion in Boletales
(BioMed Central, 2017) Castanera Andrés, Raúl; Pérez Garrido, María Gumersinda; López Varas, Leticia; Amselem, Joëlle; LaButti, Kurt; Singan, Vasanth; Lipzen, Anna; Haridas, Sajeet; Barry, Kerrie; Grigoriev, Igor V.; Pisabarro de Lucas, Gerardo; Ramírez Nasto, Lucía; Producción Agraria; Nekazaritza Ekoizpena; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Background: Coniophora olivacea is a basidiomycete fungus belonging to the order Boletales that produces brown-rot decay on dead wood of conifers. The Boletales order comprises a diverse group of species including saprotrophs and ectomycorrhizal fungi that show important differences in genome size. Results: In this study we report the 39.07-megabase (Mb) draft genome assembly and annotation of C. olivacea. A total of 14,928 genes were annotated, including 470 putatively secreted proteins enriched in functions involved in lignocellulose degradation. Using similarity clustering and protein structure prediction we identified a new family of 10 putative lytic polysaccharide monooxygenase genes. This family is conserved in basidiomycota and lacks of previous functional annotation. Further analyses showed that C. olivacea has a low repetitive genome, with 2.91% of repeats and a restrained content of transposable elements (TEs). The annotation of TEs in four related Boletales yielded important differences in repeat content, ranging from 3.94 to 41.17% of the genome size. The distribution of insertion ages of LTRretrotransposons showed that differential expansions of these repetitive elements have shaped the genome architecture of Boletales over the last 60 million years. Conclusions: Coniophora olivacea has a small, compact genome that shows macrosynteny with Coniophora puteana. The functional annotation revealed the enzymatic signature of a canonical brown-rot. The annotation and comparative genomics of transposable elements uncovered their particular contraction in the Coniophora genera, highlighting their role in the differential genome expansions found in Boletales species.