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Registros recuperados: 58
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Expresión de los genes SS y CAS en chile CM-334 inoculado con Nacobbus Aberrans y con Meloidogyne incognita. Colegio de Postgraduados
García Espinoza, Jesús Antonio.
Los esteroles son compuestos que los nematodos fitopatógenos son incapaces de sintetizar y que deben obtenerlos de su hospedante. Los genes SS y CAS codifican las enzimas escualeno sintasa y cicloartenol sintasa, respectivamente, que son clave en la síntesis de esteroles en chile. Este trabajo tuvo por objetivos, comparar la expresión de ambos genes mediante PCR en tiempo real, en la interacción compatible CM-334-Nacobbus aberrans y en la incompatible CM-334-Meloidogyne incognita, a los 2, 7, 14, 21 y 28 días después de la inoculación (ddi), así como cuantificar el número de nematodos por raíz a los 2, 7, 14, 21, 28, 35 y 42 ddi. N. aberrans completó su ciclo de vida a los 42 ddi e indujo la sobreexpresión de SS y CAS hasta dos veces más que el testigo; su...
Palavras-chave: Capsicum annuum; Cicloartenol sintasa; Escualeno sintasa; Nematodos agalladores; PCR en tiempo real; Cycloartenol synthase; Real-time PCR; Root-knot nematodes; Squalene synthase; Maestría; Fitopatología.
Ano: 2011 URL: http://hdl.handle.net/10521/492
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Transmisibilidad de aislamiento del Citrus Tristeza Virus (CTV) por áfidos y evaluación de la resistencia en cítricos tranformados con genes del CTV Colegio de Postgraduados
Loeza Kuk, Emiliano.
La sintomatología en árboles infectados por el Citrus Tristeza Virus (CTV) es influenciada por factores como la población de variantes de secuencia presentes. Esta población es modificada en la transmisión por áfidos y la evidencia sugiere que la interacción del CTV con Toxoptera citricida, incrementa la frecuencia de los aislamientos severos. Se estudió una huerta en Yucatán con una incidencia inicial de 11 árboles positivos a CTV y con tres haplotipos de aislamientos no severos, se caracterizaron los aislamientos presentes en los nuevos árboles positivos y de manera simultánea los presentes en el vector predominante (T. citricida). Después de tres años, la incidencia aumentó a 75 árboles positivos, los nuevos positivos generalmente fueron del...
Tipo: Tesis Palavras-chave: Transmisión por áfidos; SSCP; Muerte súbita de los cítricos; Plantas transgénicas; Inoculación; PCR tiempo real Aphid transmission; SSCP; Citrus sudden death; Transgenic plants; Challenging; Real-time PCR.
Ano: 2012 URL: http://hdl.handle.net/10521/1065
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Transmisibilidad de aislamiento del Citrus Tristeza Virus (CTV) por áfidos y evaluación de la resistencia en cítricos tranformados con genes del CTV Colegio de Postgraduados
Loeza Kuk, Emiliano.
La sintomatología en árboles infectados por el Citrus Tristeza Virus (CTV) es influenciada por factores como la población de variantes de secuencia presentes. Esta población es modificada en la transmisión por áfidos y la evidencia sugiere que la interacción del CTV con Toxoptera citricida, incrementa la frecuencia de los aislamientos severos. Se estudió una huerta en Yucatán con una incidencia inicial de 11 árboles positivos a CTV y con tres haplotipos de aislamientos no severos, se caracterizaron los aislamientos presentes en los nuevos árboles positivos y de manera simultánea los presentes en el vector predominante (T. citricida). Después de tres años, la incidencia aumentó a 75 árboles positivos, los nuevos positivos generalmente fueron del...
Tipo: Tesis Palavras-chave: Transmisión por áfidos; SSCP; Muerte súbita de los cítricos; Plantas transgénicas; Inoculación; PCR tiempo real; Doctorado; Fitopatología; Aphid transmission; Citrus sudden death; Transgenic plants; Challenging; Real-time PCR.
Ano: 2008 URL: http://hdl.handle.net/10521/1399
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Expresión de los genes SS y CAS en chile CM-334 inoculado con Nacobbus Aberrans y con Meloidogyne incognita. Colegio de Postgraduados
García Espinoza, Jesús Antonio.
Los esteroles son compuestos que los nematodos fitopatógenos son incapaces de sintetizar y que deben obtenerlos de su hospedante. Los genes SS y CAS codifican las enzimas escualeno sintasa y cicloartenol sintasa, respectivamente, que son clave en la síntesis de esteroles en chile. Este trabajo tuvo por objetivos, comparar la expresión de ambos genes mediante PCR en tiempo real, en la interacción compatible CM-334-Nacobbus aberrans y en la incompatible CM-334-Meloidogyne incognita, a los 2, 7, 14, 21 y 28 días después de la inoculación (ddi), así como cuantificar el número de nematodos por raíz a los 2, 7, 14, 21, 28, 35 y 42 ddi. N. aberrans completó su ciclo de vida a los 42 ddi e indujo la sobreexpresión de SS y CAS hasta dos veces más que el testigo; su...
Palavras-chave: Capsicum annuum; Cicloartenol sintasa; Escualeno sintasa; Nematodos agalladores; PCR en tiempo real; Cycloartenol synthase; Real-time PCR; Root-knot nematodes; Squalene synthase; Maestría; Fitopatología.
Ano: 2011 URL: http://hdl.handle.net/10521/492
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Importancia de las arvenses en la dispersión del amarillamiento letal del cocotero. Colegio de Postgraduados
Terán Villanueva, Nelba.
El cocotero (Cocos nucifera) uno de los cultivos tropicales de mayor importancia económica por su contribución sustancial a la seguridad alimentaria, se ha visto afectado por la enfermedad conocida como amarillamiento letal del cocotero (ALC), una de las más devastadores y que se ha localizado en todas las regiones productoras de México. El objetivo de la presente investigación fue analizar el papel que juegan las arvenses en la dispersión del ALC; para ello se realizó un registro de la flora asociada a la plantación, identificándose taxonómicamente las especies de plantas (arvenses); se hicieron análisis moleculares para detectar la presencia del fitoplasma de dicha enfermedad en las arvenses en ocho sitios dentro de la plantación. Posteriormente se...
Palavras-chave: Fitoplasma; PCR en tiempo real; Flora asociada; ALC; Myndus crudus; Phytoplasma; Real-time PCR; Associated flora; Producción Agroalimentaria en el Trópico; Maestría.
Ano: 2014 URL: http://hdl.handle.net/10521/2381
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Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit Electron. J. Biotechnol.
Taski-Ajdukovic,Ksenija; Nagl,Nevena; Kovacev,Lazar; Curcic,Zivko; Danojevic,Dario.
Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the genomic level. Since recent research efforts have focused on the molecular response of the plant in order to identify water deficit inducible genes, the aim of this investigation was to develop qRT-PCR methodology for the quantification of gene expression in sugar beet under conditions of water deficiency in vitro. Sugar beet genotypes, selected for different response to water deficit, were grown and multiplied in vitro. Axilary shoots were placed on...
Tipo: Journal article Palavras-chave: Beta vulgaris; Drought; In vitro; Real-time PCR.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000600010
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Effect of cryopreservation on the efficiency of exogenous gene, genetic transformation and expression level of Arabidopsis thaliana Electron. J. Biotechnol.
Li,Zhongai; Du,Yaqiong; Wang,Zicheng.
Background: Cryopreservation refers to the storage of a living organism at ultra-low-temperature for long-term preservation of plant germplasm. The effect of cryopreservation on the efficiency of exogenous gene genetic transformation and expression level were studied herein. In this work, transgenic Arabidopsis thaliana were successfully conserved in vitro by cryopreservation methods. Results: The effects of osmotic stress due to cryoprotectants during pretreatment and of storage at -196ºC on the stability, the efficiency of genetic transformation and the expression level of exogenous gene were analyzed in Arabidopsis. The results showed that there had not any significant increasing in the efficiency of genetic transformation after cryopreservation, and...
Tipo: Journal article Palavras-chave: Arabidopsis thaliana; Cryopreservation; Real-time PCR; Transformation efficiency.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600012
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A real-time PCR genotyping assay to detect FAD2A SNPs in peanuts (Arachis hypogaea L.) Electron. J. Biotechnol.
Barkley,Noelle A; Wang,Ming Li; Pittman,Roy N.
The high oleic (C18:1) phenotype in peanuts has been previously demonstrated to result from a homozygous recessive genotype (ol1ol1ol2ol2) in two homeologous fatty acid desaturase genes (FAD2A and FAD2B) with two key SNPs. These mutant SNPs, specifically G448A in FAD2A and 442insA in FAD2B, significantly limit the normal function of the desaturase enzyme activity which converts oleic acid into linoleic acid by the addition of a second double bond in the hydrocarbon chain. Previously, a genotyping assay was developed to detect wild type and mutant alleles in FAD2B. A real-time PCR assay has now been developed to detect wild type and mutant alleles (G448A) in FAD2A using either seed or leaf tissue. This assay was demonstrated to be applicable for the...
Tipo: Journal article Palavras-chave: Fatty acid composition; Gas chromatography; Peanut (Arachis hypogaea L.); Real-time PCR; SNP genotyping.
Ano: 2011 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100009
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Molecular identification of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila Genet. Mol. Biol.
Mota,Adolfo Jose; Back-Brito,Graziella Nuernberg; Nobrega,Francisco G..
Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila. As result, these species are often misidentified. In this work, we established a reliable method for the identification/differentiation of these species. Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region. This assay provides a new tool for pathogen identification and for epidemiological, drug resistance and virulence studies of these organisms.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Candida palmioleophila; Debaryomyces hansenii; Differential identification; Pichia guilliermondii; Real-time PCR.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000100017
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Quantitative real-time PCR for the clinical detection of Helicobacter pylori Genet. Mol. Biol.
Ribeiro,Marcelo Lima; Ecclissato,Christina Cunha; Mattos,Ricardo Gabriel; Mendonca,Sergio; Pedrazzoli Jr.,José.
Accurate diagnosis of Helicobacter pylori infection is very important in both clinical practice and research. We evaluated the sensitivity of real-time PCR (RT-PCR) for the detection and quantification of Helicobacter pylori using DNA from 91 human gastric biopsy samples divided into three groups: 46 biopsies from untreated patients who according to the references methods were considered H. pylori-negative (group A); 35 biopsies from patients previously treated against H. pylori and considered to be cured by "gold standard" tests (group B); and 10 biopsies from patients H. pylori-positive by all available methods (group C). The sensitivity of the RT-PCR assay was higher than that of standard methods. Of the 81 patients considered to be uninfected according...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Diagnostic methods; Helicobacter pylori; Real-time PCR.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300022
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Expression profile of genes associated with mastitis in dairy cattle Genet. Mol. Biol.
Fonseca,Isabela; Silva,Priscila Vendramini; Lange,Carla Christine; Guimarães,Marta F.M.; Weller,Mayara Morena Del Cambre Amaral; Sousa,Katiene Régia Silva; Lopes,Paulo Sávio; Guimarães,José Domingos; Guimarães,Simone E.F..
In order to characterize the expression of genes associated with immune response mechanisms to mastitis, we quantified the relative expression of the IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ and TNF-α genes in milk cells of healthy cows and cows with clinical mastitis. Total RNA was extracted from milk cells of six Black and White Holstein (BW) cows and six Gyr cows, including three animals with and three without mastitis per breed. Gene expression was analyzed by real-time PCR. IL-10 gene expression was higher in the group of BW and Gyr cows with mastitis compared to animals free of infection from both breeds (p < 0.05). It was also higher in BW Holstein animals with clinical mastitis (p < 0.001), but it was not significant when Gyr cows with and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cytokine; Gene expression; Immune response; Real-time PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000400018
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Simultaneous quantitative assessment of circulating cell-free mitochondrial and nuclear DNA by multiplex real-time PCR Genet. Mol. Biol.
Xia,Peng; Radpour,Ramin; Zachariah,Rebecca; Fan,Alex Xiu Cheng; Kohler,Corina; Hahn,Sinuhe; Holzgreve,Wolfgang; Zhong,Xiao Yan.
Quantification of circulating nucleic acids in plasma and serum could be used as a non-invasive diagnostic tool for monitoring a wide variety of diseases and conditions. We describe here a rapid, simple and accurate multiplex real-time PCR method for direct synchronized analysis of circulating cell-free (ccf) mitochondrial (mtDNA) and nuclear (nDNA) DNA in plasma and serum samples. The method is based on one-step multiplex real-time PCR using a FAM-labeled MGB probe and primers to amplify the mtDNA sequence of the ATP 8 gene, and a VIC-labeled MGB probe and primers to amplify the nDNA sequence of the glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) gene, in plasma and serum samples simultaneously. The efficiencies of the multiplex assays were measured in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Circulating cell-free DNA; Mitochondrial DNA; Nuclear DNA; Real-time PCR; Quantitative PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000100003
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NFAT5 is differentially expressed in Sprague-Dawley rat tissues in response to high salt and high fructose diets Genet. Mol. Biol.
Herman,Braden A.; Ferguson,Kaylee M.; Fernandez,Jared V.B.; Kauffman,Samantha; Spicher,Jason T.; King,Rachel J.; Halterman,Julia A..
Abstract Current diets contain an increasing amount of salt and high fructose corn syrup, but it remains unclear as to how dietary salt and fructose affect organ function at the molecular level. This study aimed to test the hypothesis that consumption of high salt and fructose diets would increase tissue-specific expression of two critical osmotically-regulated genes, nuclear factor of activated T-cells 5 (NFAT5) and aldose reductase (AR). Fifty Sprague-Dawley rats were placed on a control, 4% NaCl, 8% NaCl, or 64% fructose diet for eight weeks. Fourteen different tissue samples were harvested and snap-frozen, followed by RNA purification, cDNA synthesis, and NFAT5 and AR gene expression quantification by real-time PCR.Our findings demonstrate that NFAT5...
Tipo: Info:eu-repo/semantics/article Palavras-chave: NFAT5; Aldose reductase; Salt; Fructose; Real-time PCR.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572019000300452
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Caractérisation et quantification moléculaires de l’écosystème microbien d’altération du saumon cru et des crevettes cuites ArchiMer
Mace, Sabrina.
Raw salmon and cooked shrimp sold in supermarket are usually found under vacuum or CO2 modified atmosphere packaging (MAP). This type of packaging inhibit growth of some bacteria, therefore, CO2 tolerant bacteria dominate spoilage microbiota of these products. The aim of this study is to improve knowledge about bacteria involved in spoilage. Spoilage microbial ecosystems of raw salmon and tropical cooked shrimp were described using classic miocrobiological and molecular methods like PCR-TTGE and pyrosequencing. Lactic acid bacteria, notably Lactococcus piscium, Carnobacterium maltaromaticum, Carnobacterium divergens, Lactobacillus sakei, and Gram negative bacteria like Photobacterium phosphoreum and Enterobacteriaceae (Serratia spp.) has been described as...
Tipo: Text Palavras-chave: Produits de la mer; Microbiote; PCR-TTGE; Pyroséquençage; Potentiel d’altération; P. phosphoreum; PCR en temps-réel; Seafood products; Microbiota; PCR-TTGE; Pyrosequencing; Spoilage potential; P. phosphoreum; Real-time PCR.
Ano: 2013 URL: https://archimer.ifremer.fr/doc/00166/27773/25963.pdf
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Quantification des bactéries histaminogènes et maîtrise de la formation d'histamine dans les produits marins par biopréservation ArchiMer
Podeur, Gaetan.
Histamine-producing bacteria were isolated from naturally contaminated seafood, some of responsible for histamine-poisoning. At the same time, lactic acid bacteria were isolated on the same products and clustered according to their inhibitory activity against 4 histamine-producing species. Most of the strains were identified as Lactobacillus fuchuensis or Lactobacillus sakei. Twelve lactic acid bacteria were selected for challenge-test on canned, cooked or smoked tuna. Challenge-test combined with sensory analysis performed on cooked vacuum-packed tuna against Morganella psychrotolerans and Morganella morganii, demonstrated an important inhibitory effect on growth and production of histamine with L. sakei CNCM I-4707. Bacterial count was reduced by 3 to 5...
Tipo: Text Palavras-chave: Biopréservation; Thon; Bactérie lactique; Intoxication histaminique; Morganella morganii; Morganella psychrotolerans; Lactobacillus sakei; Analyse sensorielle; PCR en temps réel; Étape d'enrichissement; RT-qPCR; Biopreservation; Tuna; Lactic acid bacteria; Scombroid poisoning; Morganella morganii; Morganella psychrotolerans; Lactobacillus sakei; Sensory analysis; Real-time PCR; Enrichment step; RT-qPCR.
Ano: 2014 URL: https://archimer.ifremer.fr/doc/00241/35203/33709.pdf
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Identification of the autophagy pathway in a mollusk bivalve, Crassostrea gigas ArchiMer
Picot, Sandy; Faury, Nicole; Arzul, Isabelle; Chollet, Bruno; Renault, Tristan; Morga, Benjamin.
The Pacific oyster, Crassostrea gigas, is a mollusk bivalve commercially important as a food source. Pacific oysters are subjected to stress and diseases during culture. The autophagy pathway is involved in numerous cellular processes, including responses to starvation, cell death, and microorganism elimination. Autophagy also exists in C. gigas, and plays a role in the immune response against infections. Although this process is well-documented and conserved in most animals, it is still poorly understood in mollusks. To date, no study has provided a complete overview of the molecular mechanism of autophagy in mollusk bivalves. In this study, human and yeast ATG protein sequences and public databases (Uniprot and NCBI) were used to identify protein members...
Tipo: Text Palavras-chave: Autophagy; Autophagy related; Crassostrea gigas; Data mining; Immunohistochemistry; Real-time PCR; Western blot.
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00606/71785/70270.pdf
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Recent innovation in microbial source tracking using bacterial real-time PCR markers in shellfish ArchiMer
Mauffret, Aourell; Mieszkin, Sophie; Morizur, Mael; Alfiansah, Yustian Rovi; Lozach, Solen; Gourmelon, Michele.
We assessed the capacity of real-time PCR markers to identify the origin of contamination in shellfish. Oyster, cockles or clams were either contaminated with fecal materials and host-associated markers designed from Bacteroidales or Catellicoccus marimammalium 16S RNA genes were extracted from their intravalvular liquid, digestive tissues or shellfish flesh. Extraction of bacterial DNA from the oyster intravalvular liquid with FastDNA spin kit for soil enabled the selected markers to be quantified in 100% of artificially contaminated samples, and the source of contamination to be identified in 13 out of 38 naturally contaminated batches from European Class B and Class C areas. However, this protocol did not enable the origin of the contamination to be...
Tipo: Text Palavras-chave: Microbial source tracking; Shellfish; Bacteroidales; Real-time PCR; Intravalvular liquid; Digestive tissues.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00137/24776/25188.pdf
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Development of TaqMan real-time PCR assays for monitoring Vibrio harveyi infection and a plasmid harbored by virulent strains in European abalone Haliotis tuberculata aquaculture ArchiMer
Schikorski, David; Renault, Tristan; Paillard, Christine; Bidault-toffin, A.; Tourbiez, Delphine; Saulnier, Denis.
The Gram-negative bacterium Vibrio harveyi is known to be highly pathogenic for the European abalone Haliotis tuberculata, which is a gastronomically important marine gastropod with a high commercial value. Since 1998, some particular bacterial strains are described as implicated in recurrent mortality outbreaks in French farm and field stocks of abalone. Recently, a 9.6kb plasmid named pVCR1, was shown to be harbored by one highly V. harveyi virulent ORM4 strain suggesting its involvement in virulence phenotype. Thus, we have developed in the present study two TaqMan real-time PCR assays allowing to (i) rapidly and specifically detect, by a duplex procedure and in less than 2 h, both V. harveyi and the presence of plasmid pVCR1 from unidentified bacterial...
Tipo: Text Palavras-chave: Vibrio harveyi; Haliotis tuberculata; Real-time PCR; Plasmid; Marine pathogen; Molecular diagnostic.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00124/23540/21383.pdf
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Historical records from dated sediment cores reveal the multidecadal dynamic of the toxic dinoflagellate Alexandrium minutum in the Bay of Brest (France) ArchiMer
Klouch, Khadidja; Schmidt, Sabine; Andrieux-loyer, Francoise; Le Gac, Mickael; Hervio-heath, Dominique; Qui-minet, Zujaila Nohemi; Quere, Julien; Bigeard, Estelle; Guillou, Laure; Siano, Raffaele.
The multiannual dynamic of the cyst-forming and toxic marine dinoflagellate Alexandrium minutum was studied over a time scale of about 150 years by a paleoecological approach based on ancient DNA (aDNA) quantification and cyst revivification data obtained from two dated sediment cores of the Bay of Brest (Brittany, France). The first genetic traces of the species presence in the study area dated back to 1873 ± 6. Specific aDNA could be quantified by a newly-developed real-time PCR assay in the upper core layers, in which the germination of the species (in up to 17–19 year-old sediments) was also obtained. In both cores studied, our quantitative paleogenetic data showed a statistically significant increasing trend in the abundance of A. minutum ITS1 rDNA...
Tipo: Text Palavras-chave: Paleoecology; Ancient DNA; Real-time PCR; Harmful algal blooms (HAB); Dinoflagellates; Coastal ecology.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00334/44516/44212.pdf
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Etude des interactions hôte/virus chez l’huître creuse Crassostrea gigas et son virus Ostreid herpesvirus 1 ArchiMer
Segarra, Amelie.
Ostreid herpesvirus type 1 (OsHV-1) can be considered one of the major infectious agents in Pacific oysters, Crassostrea gigas, in France and all around the world. Differences in terms of mortality among infected animals were reported suggesting a genetic basis of the susceptibility to the OsHV-1 infection. In this context, the main objective of this thesis was to understand the interactions between Pacific oysters and OsHV-1, in particular, the molecular basis of the viral cycle. Results showed that the virus is able to replicate in the host regardless of its stage of development or its susceptibility. However, multiplication kinetics is faster in the most susceptible individuals compared to less susceptible ones. After an active replication phase, it...
Tipo: Text Palavras-chave: Bivalves; Huître creuse; Crassostrea gigas; Mortalités; OsHV-1; Gènes viruax; Interaction hôte-virus; Transcriptome; Sensibilité; PCR en temps réel; Bivalve; Pacific oyster; Crassostrea ggias; Mortality; OsHV-1; Viral gene; Interaction host-virus; Transcriptome; Susceptibility; Real-time PCR.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00252/36297/34826.pdf
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