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1.
Sci Rep ; 7(1): 6749, 2017 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-28751733

RESUMO

The tissue-specific etiology of aging and stress has been elusive due to limitations in data processing of current techniques. Despite that many techniques are high-throughput, they usually use singular features of the data (e.g. whole fluorescence). One technology at the nexus of fluorescence-based screens is large particle flow cytometry ("biosorter"), capable of recording positional fluorescence and object granularity information from many individual live animals. Current processing of biosorter data, however, do not integrate positional information into their analysis and data visualization. Here, we present a bioanalytical platform for the quantification of positional information ("longitudinal profiling") of C. elegans, which we posit embodies the benefits of both high-throughput screening and high-resolution microscopy. We show the use of these techniques in (1) characterizing distinct responses of a transcriptional reporter to various stresses in defined anatomical regions, (2) identifying regions of high mitochondrial membrane potential in live animals, (3) monitoring regional mitochondrial activity in aging models and during development, and (4) screening for regulators of muscle mitochondrial dynamics in a high-throughput format. This platform offers a significant improvement in the quality of high-throughput biosorter data analysis and visualization, opening new options for region-specific phenotypic screening of complex physiological phenomena and mitochondrial biology.


Assuntos
Envelhecimento/genética , Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Ensaios de Triagem em Larga Escala , Mitocôndrias/metabolismo , Dinâmica Mitocondrial/genética , Envelhecimento/metabolismo , Animais , Animais Geneticamente Modificados , Caenorhabditis elegans/crescimento & desenvolvimento , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/ultraestrutura , Proteínas de Caenorhabditis elegans/metabolismo , Escherichia coli/crescimento & desenvolvimento , Citometria de Fluxo/métodos , Regulação da Expressão Gênica no Desenvolvimento , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Intestinos/ultraestrutura , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Potencial da Membrana Mitocondrial/fisiologia , Mitocôndrias/ultraestrutura , Músculos/metabolismo , Músculos/ultraestrutura , Especificidade de Órgãos , Faringe/crescimento & desenvolvimento , Faringe/metabolismo , Faringe/ultraestrutura , Células Receptoras Sensoriais/citologia , Células Receptoras Sensoriais/metabolismo , Transcrição Gênica , Proteína Vermelha Fluorescente
2.
PLoS One ; 10(6): e0127539, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26062090

RESUMO

Climatic changes impact fruit tree growth and severely limit their production. Investigating the tree ability to cope with environmental variations is thus necessary to adapt breeding and management strategies in order to ensure sustainable production. In this study, we assessed the genetic parameters and genotype by environment interaction (GxE) during the early tree growth. One hundred and twenty olive seedlings derived from the cross 'Olivière' x 'Arbequina' were examined across two sites with contrasted environments, accounting for ontogenetic trends over three years. Models including the year of growth, branching order, environment, genotype effects, and their interactions were built with variance function and covariance structure of residuals when necessary. After selection of a model, broad sense heritabilities were estimated. Despite strong environmental effect on most traits, no GxE was found. Moreover, the internal structure of traits co-variation was similar in both sites. Ontogenetic growth variation, related to (i) the overall tree form and (ii) the growth and branching habit at growth unit scale, was not altered by the environment. Finally, a moderate to strong genetic control was identified for traits at the whole tree scale and at internode scale. Among all studied traits, the maximal internode length exhibited the highest heritability (H2 = 0.74). Considering the determinant role of this trait in tree architecture and its stability across environments, this study consolidates its relevance for breeding.


Assuntos
Interação Gene-Ambiente , Modelos Genéticos , Olea/genética , Olea/crescimento & desenvolvimento , Melhoramento Vegetal , Plântula/genética , Plântula/crescimento & desenvolvimento
3.
PLoS One ; 8(5): e62831, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23690957

RESUMO

One of the challenge fruit growers are facing is to balance between tree production and vegetative growth from year to year. To investigate the existence of genetic determinism for reproductive behaviour in olive tree, we studied an olive segregating population derived from a cross between 'Olivière' and 'Arbequina' cultivars. Our strategy was based on (i) an annual assessment of individual trees yield, and (ii) a decomposition of adult growth units at the crown periphery into quantitative variables related to both flowering and fruiting process in relation to their growth and branching. Genetic models, including the year, genotype effects and their interactions, were built with variance function and correlation structure of residuals when necessary. Among the progeny, trees were either 'ON' or 'OFF' for a given year and patterns of regular vs. irregular bearing were revealed. Genotype effect was significant on yield but not for flowering traits at growth unit (GU) scale, whereas the interaction between genotype and year was significant for both traits. A strong genetic effect was found for all fruiting traits without interaction with the year. Based on the new constructed genetic map, QTLs with small effects were detected, revealing multigenic control of the studied traits. Many were associated to alleles from 'Arbequina'. Genetic correlations were found between Yield and Fruit set at GU scale suggesting a common genetic control, even though QTL co-localisations were in spe`cific years only. Most QTL were associated to flowering traits in specific years, even though reproductive traits at GU scale did not capture the bearing status of the trees in a given year. Results were also interpreted with respect to ontogenetic changes of growth and branching, and an alternative sampling strategy was proposed for capturing tree fruiting behaviour. Regular bearing progenies were identified and could constitute innovative material for selection programs.


Assuntos
Flores/genética , Frutas/genética , Olea/genética , Locos de Características Quantitativas/genética , Mapeamento Cromossômico , Cruzamentos Genéticos , Genótipo , Modelos Genéticos , Olea/crescimento & desenvolvimento
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