23 research outputs found

    From methylome to integrative analysis of tissue specificity

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    International audienceDNA methylation is the most studied epigenetic mark in both plants and animals. The gold standard for assaying genome-wide DNA methylation at single-base resolution is whole-genome bisulfite sequencing (WGBS). Here, we describe an improved procedure for WGBS and original bioinformatic workflows applied to unravel tissue-specific variations of the methylome in relation to gene expression and accumulation of secondary metabolites in the medicinal plant Catharanthus roseus

    Xylem sap proteomics

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    Xylem sap proteomic

    The constitutive phenolic composition of two Malus x domestica genotypes is not responsible for their contrasted susceptibilities to fire blight

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    Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699International audienceApple (Malus x domestica) leaf phenolics, in particular the three major dihydrochalcones, phloridzin, trilobatin and sieboldin, were studied for their potential contribution to the resistance to Erwinia amylovora, the causative agent of fire blight. The constitutive phenolic compositions of part of a progeny resulting from a cross between two apple genotypes, displaying contrasted susceptibilities to fire blight, were investigated by RP-HPLC-DAD. With regards to the major dihydrochalcones, offsprings were found to display the parent profile: either phloridzin alone or a combination of phloridzin, trilobatin and sieboldin. Despite a strong antioxidant activity that could interfere with the oxidative burst triggered by the bacteria, sieboldin did not segregate with resistance even though it was strongly associated with a high constitutive antioxidant capacity of leaf extracts. Several hydroxycinnamic acids and flavonols were identified in the progeny but profiles were very similar. However, concentrations of several flavonols positively correlated with resistance in the genotypes that possessed trilobatin and sieboldin. The major dihydrochalcones that represent more than 200 mg/g of leaf DW are not directly responsible for resistance to fire blight in this system. In addition, a high constitutive antioxidant activity is unlikely to be sufficient to block bacterial sprea

    An Erwinia amylovora inducible promoter for intragenic improvement of apple fire blight resistance

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    Intragenesis is an important alternative to transgenesis to produce modified plants 27 containing native DNA only. A key point to develop such a strategy is the availability of 28 regulatory sequences controlling the expression of the gene of interest. With the aim of 29 finding apple gene promoters either inducible by the fire blight pathogen Erwinia amylovora 30 (Ea) or moderately constitutive, we focused on polyphenoloxidase genes (PPO). These genes 31 encode oxidative enzymes involved in many physiological processes and have been 32 previously shown to be up-regulated during the Ea infection process. We found ten PPO and 33 two PPO-like sequences in the apple genome and characterized the promoters of MdPPO16 34 (pPPO16) and MdKFDV02 PPO-like (pKFDV02) for their potential as Ea-inducible and low-35 constitutive regulatory sequences respectively. Expression levels of reporter genes fused to 36 these promoters and transiently or stably expressed in apple were quantified after various 37 treatments. Unlike pKFDV02 which displayed a variable activity, pPPO16 allowed a fast and 38 strong expression of transgenes in apple following Ea infection in a Type 3 Secretion System 39 dependent manner. Altogether our results indicate that pKFDV02 did not keep its promises as 40 a constitutive and weak promoter whereas pPPO16, the first Ea-inducible promoter cloned 41 from apple, can be a useful component of intragenic strategies to create fire blight resistant 42 apple genotypes

    Transcriptional reprogramming and phenotypical changes associated with growth of Xanthomonas campestris pv. campestris in cabbage xylem sap

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    Xylem sap (XS) is the first environment that xylem phytopathogens meet in planta during the early infection steps. Xanthomonas campestris pv. campestris (Xcc), the causative agent of Brassicaceae black rot, colonizes the plant xylem vessels to ensure its multiplication and dissemination. Besides suppression of plant immunity, Xcc has to adapt its metabolism to exploit plant-derived nutrients present in XS. To study Xcc behaviour in the early infection steps, we used cabbage XS to analyse bacterial growth. Mineral and organic composition of XS were determined. Significant growth of Xcc in XS was allowed by the rapid catabolism of amino acids, sugars and organic acids, and it was accompanied by the formation of biofilm-like structures. Transcriptome analysis of Xcc cultivated in XS using cDNA microarrays revealed a XS-specific transcriptional reprogramming compared to minimal or rich media. More specifically, up-regulation of genes encoding transporters such as TonB-dependent transporters (TBDTs), that could be associated with nutrient acquisition and detoxification, was observed. In agreement with the aggregation phenotype, expression of genes important for twitching motility and adhesion was up-regulated in XS. Taken together, our data show specific responses of Xcc to colonization of cabbage XS that could be important for the pathogenesis process and establish XS as a model medium to study mechanisms important for the early infection events
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