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    ABSTRACT: While steady-state (13)C metabolic flux analysis is a powerful method for deducing multiple fluxes in the central metabolic network of heterotrophic and mixotrophic plant tissues, it is also time-consuming and technically challenging. Key steps in the design and interpretation of steady-state (13)C labeling experiments are illustrated with a generic protocol based on applications to plant cell suspension cultures.
    No preview · Article · Jan 2014 · Methods in molecular biology (Clifton, N.J.)
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    ABSTRACT: The analysis of stable isotope incorporation following feeding of (13)C-labeled precursors to plant tissues provides the constraints necessary for metabolic flux analysis. This protocol describes the use of one-dimensional (1)H and (13)C nuclear magnetic resonance spectroscopy for the quantification of (13)C enrichments and isotopomer abundances in mixtures of metabolites or hydrolyzed biomass components.
    No preview · Article · Jan 2014 · Methods in molecular biology (Clifton, N.J.)
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    ABSTRACT: Inoculations with the saprophytic fungus Trichoderma spp. are now extensively used both to promote plant growth and suppress disease development. The underlying mechanisms for both roles have yet to be fully described so that the use of Trichoderma could be optimised. We here show that Trichoderma asperelloides effects include the manipulation of host NO production. NO was rapidly formed in Arabidopsis roots in response to soil borne necrotrophic pathogen Fusarium. oxysporum and persisted for ~1 h but is only transiently produced ( ~ 10 min) when roots interact with T. asperelloides (T203). However, inoculation of F. oxysporum infected roots with T. asperelloides suppressed F. oxysporum initiated NO production. A transcriptional study of 78 NO modulated genes indicated most genes were suppressed by single and combinational challenge with F. oxysporum or T. asperelloides. Only two F. oxysporum induced genes were suppressed by T. asperelloides inoculation undertaken either 10 min prior to or post pathogen infection; a concanavlin A-like lectin protein kinase (At4g28350) and the receptor-like protein, RLP30. Thus T. asperelloides can actively suppress NO production elicited by F. oxysporum and impacts on the expression of some genes reported to be NO-responsive. Of particular interest was the reduced expression of receptor-like genes which may be required for F. oxysporum dependent necrotrophic disease development.
    Full-text · Article · Nov 2013 · Molecular Plant-Microbe Interactions
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