5 resultados para Molecular Analysis

em Universidad Politécnica de Madrid


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The endo-β-mannanase (MAN) family is represented in the Arabidopsis genome by eight members, all with canonical signal peptides and only half of them being expressed in germinating seeds. The transcripts of these genes were localized in the radicle and micropylar endosperm (ME) before radicle protrusion and this expression disappears as soon as the endosperm is broken by the emerging radicle tip. However, only three of these MAN genes, AtMAN5, AtMAN7 and especially AtMAN6 influence the germination time (t50) as assessed by the analysis of the corresponding knock-out lines. The data suggest a possible interaction between embryo and ME regarding the role of MAN during the Arabidopsis germination process.

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Bread wheat quality constitutes a key trait for the demands of the baking industry as well as the broad consumer preferences. The role of the low molecular weight glutenin subunits (LMW-GS) with regard to bread quality is so far not well understood owing to their genetic complexity and to the use of different nomenclatures and standards for the LMW-GS assignment by different research groups, which has made difficult the undertaking of association studies between genotypes and bread quality. The development of molecular markers to carry out genetic characterization and allele determination is demanding. Nowadays, the most promising LMW gene marker system is based on PCR and high resolution capillary electrophoresis for the simultaneous analysis of the complete multigene family. The molecular analysis of the bread wheat Glu-B3 locus in F2 and F4:6 populations expressed the expected one-locus Mendelian segregation pattern, thus validating the suitability of this marker system for the characterization of LMW-GS genes in segregating populations, allowing for the successful undertaking of studies related to bread-making quality. Moreover, the Glu-B3 allele characterization of standard cultivars with the molecular marker system has revealed its potential as a complementary tool for the allelic determination of this complex multigene family.

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In prokaryotes, nickel is an essential element participating in the structure of enzymes involved in multiple cellular processes. Nickel transport is a challenge for microorganisms since, although essential, high levels of this metal inside the cell are toxic. For this reason, bacteria have developed high-affinity nickel transporters as well as nickel-specific detoxification systems. Ultramafic soils, and soils contaminated with heavy metals are excellent sources of nickel resistant bacteria. Molecular analysis of strains isolated in the habitats has revealed novel genetic systems involved in adaptation to such hostile conditions.

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La vid silvestre se considera como el ancestro autóctono de las vides cultivadas y una enorme reserva genética en peligro de extinción. La prospección llevada a cabo entre 2003 y 2004 permitió catalogar 51 localizaciones de vides silvestres españolas, la mayoría de ellas ubicadas en riberas de ríos. Estos ejemplares se incluyeron en el Banco de Germoplasma de la Finca "El Encín" (BGVCAM - Alcalá de Henares, Madrid, España). En primer lugar, se caracterizó la cantidad y la distribución de su diversidad genética utilizando 25 loci empleando microsatélites nucleares (SSR). Hemos analizado también la posible coexistencia en el hábitat natural de vides silvestres con vides cultivadas naturalizadas y portainjertos. De este modo, los análisis fenotípicos y genéticos identificaron el 19% de las muestras recogidas como derivadas de genotipos cultivados, siendo, o bien vides cultivadas naturalizadas o genotipos híbridos derivados de cruces espontáneos entre vides silvestres y cultivadas. La diversidad genética de las poblaciones de vides silvestres fue similar a la observada en el grupo de las cultivadas. El análisis molecular mostró que el germoplasma de cultivadas y silvestres es genéticamente divergente con bajo nivel de introgresión. Se ha identificado cuatro grupos genéticos, con dos de ellos fundamentalmente representados por los genotipos de vides cultivadas y dos por las accesiones silvestres. El análisis de los vínculos genéticos entre las vides silvestres y cultivadas podría sugerir una contribución genética de las accesiones silvestres españolas a las actuales variedades occidentales. En segundo lugar, se realizó un profundo estudio morfológico "ex situ " y se contrastaron con los resultados de la caracterización realizada en 182 variedades comerciales españolas de la misma colección. Todos los individuos silvestres mostraron diferencias morfológicas con Vitis vinifera L subsp. vinifera, pero no se encontraron diferencias significativas dentro Vitis vinifera L. subsp. sylvestris, ni por localización geográfica ni por sexo. Los resultados de este estudio describen las principales características morfológicas de las vides silvestres españolas y sus rasgos diferenciales con su pariente cultivada. Por último, se analizó la composición antociánica presente en 21 accesiones de vides silvestres de la Península Ibérica conservadas en el BGVCAM de la Finca "El Encín" y seleccionadas basándose en diferencias ampelográficas y caracterización molecular. La concentración de antocianinas es similar la encontrad en vides cultivadas con destino a la vinificación. Las accesiones estudiadas mostraron una variabilidad considerable en su perfil antociánico y fue posible distinguir varios grupos. Sin embargo, la presencia de material silvestre con perfiles antociánicos poco comunes o inexistentes en variedades españolas, sugiere que la variabilidad genética relacionada con antocianinas en poblaciones españolas de vides silvestres podría ser más alta que la de variedades cultivadas comúnmente consideradas de origen español. ABSTRACT The wild grapevine is considered an autochthonous relative of cultivated vines and a huge gene pool endangered in Europe. Prospecting carried out between 2003 and 2004 enabled to inventory 51 Spanish sites with wild grapevines, most of them located near rivers. These individuals were grafted in the collection of "El Encín" (BGVCAM - Alcalá de Henares, Madrid, Spain). Firstly, werw characterized the amount and distribution of their genetic diversity using 25 nuclear SSR loci. We have also analysed the possible coexistence in the natural habitat of wild grapevines with naturalized grapevine cultivars and rootstocks. In this way, phenotypic and genetic analyses identified 19% of the collected samples as derived from cultivated genotypes, being either naturalized cultivars or hybrid genotypes derived from spontaneous crosses between wild and cultivated grapevines. The genetic diversity of wild grapevine populations was similar than that observed in the cultivated group. The molecular analysis showed that cultivated germplasm and wild germplasm are genetically divergent with low level of introgression. We identified four genetic groups, with two of them fundamentally represented among cultivated genotypes and two among wild accessions. The analyses of genetic relationships between wild and cultivated grapevines could suggest a genetic contribution of wild accessions from Spain to current Western cultivars. Secondly, a morphological study was done "ex situ" and were compared with data from 182 Spanish commercial cultivars grown in the same collection. All wild individuals showed morphological differences with Vitis vinifera L. ssp. vinifera but no significant differences were found within Vitis vinifera L subsp. sylvestris neither by geographic origin nor by sex. A pattern with the main characteristics of Spanish wild grapevines is suggested. Ultimately, were investigated the anthocyanin composition of 21 mostly Spanish wild grapevine accessions preserved at BGVCAM "El Encín" and selected in consideration of observed ampelographic differences and molecular characterization. Total anthocyanin concentration was similar to that found in winegrape cultivars. The accessions studied showed considerable variability in their anthocyanin fingerprints and it was possible to distinguish several groups, similar to previous reports on the anthocyanin fingerprint of winegrapes. The anthocyanin composition of wild grapevine accessions was similar to that of cultivated grapes. Nevertheless, the presence of wild accessions with anthocyanin fingerprints uncommon or nonexistent in Spanish cultivated varieties suggests that the genetic variability related to anthocyanins in Spanish wild grapevine populations may be higher than that of cultivated varieties commonly considered of Spanish origin.

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One of the biggest challenges that software developers face is to make an accurate estimate of the project effort. Radial basis function neural networks have been used to software effort estimation in this work using NASA dataset. This paper evaluates and compares radial basis function versus a regression model. The results show that radial basis function neural network have obtained less Mean Square Error than the regression method.