Ectopic expression of PtaRHE1, encoding a poplar RING-H2 protein with E3 ligase activity, alters plant development and induces defence-related responses

RING (really interesting new gene)-H2 domain-containing proteins are widely represented in plants and play important roles in the regulation of many developmental processes as well as in plant–environment interactions. In the present report, experiments were performed to unravel the role of the poplar gene PtaRHE1, coding for a RING-H2 protein. In vitro ubiquitination assays indicate a functional E3 ligase activity for PtaRHE1 with the specific E2 ubiquitin-conjugating enzyme UbcH5a. The overexpression of PtaRHE1 in tobacco resulted in a pleiotropic phenotype characterized by a curling of the leaves, the formation of necrotic lesions on leaf blades, growth retardation, and a delay in floral transition. The plant gene expression response to PtaRHE1 overexpression provided evidence for the up-regulation of defence- and/or programmed cell death-related genes. Moreover, genes coding for WRKY transcription factors as well as for mitogen-activated protein kinases, such as wound-induced protein kinase (WIPK), were also found to be induced in the transgenic lines as compared with the wild type. In addition, histochemical β-glucuronidase staining showed that the PtaRHE1 promoter is induced by plant pathogens and by elicitors such as salicylic acid and cellulase. Taken together, these results suggest that the E3 ligase PtaRHE1 plays a role in the ubiquitination-mediated regulation of defence response, possibly by acting upstream of WIPK and/or in the activation of WRKY factors

Can we identify wild-born salmon from parentage assignment data? A case study in the Garonne-Dordogne rivers salmon restoration programme in France

Parentage assignment with genomic markers provides an opportunity to monitor salmon restocking programs. Most of the time, it is used to study the fate of hatchery-born fish in those programs, as well as the genetic impacts of restocking. In such analyses, only fish that are assigned to their parents are considered. In the Garonne-Dordogne river basin in France, native salmon have disappeared, and supportive breeding is being used to try to reinstate a self-sustained population. It is therefore of primary importance to assess the numbers of wild-born returning salmon, which could appear as wrongly assigned or not assigned, depending on the power of the marker set and on the size of the mating plan. We used the genotypes at nine microsatellites of the 5800 hatchery broodstock which were used from 2008 to 2014, and of 884 upstream migrating fish collected from 2008 to 2016, to assess our ability to identify wild-born salmon. We simulated genotypes of hatchery fish and wild-born fish and assessed how they were identified by the parentage assignment software Accurassign. We showed that 98.7% of the fish assigned within the recorded mating plan could be considered hatchery fish, while 93.3% of the fish in other assignment categories (assigned out of the mating plan, assigned to several parent pairs, not assigned) could be considered wild-born. Using a Bayesian approach, we showed that 31.3% of the 457 upstream migrating fish sampled from 2014 to 2016 were wild-born. This approach is thus efficient to identify wild-born fish in a restoration program. It remains dependent on the quality of the recording of the mating plan, which we showed was rather good (<5% mistakes) in this program. To limit this potential dependence, an increase in the number of markers genotyped (17 instead of 9) is now being implemented

Development of a candidate reference material for adventitious virus detection in vaccine and biologicals manufacturing by deep sequencing.

Unbiased deep sequencing offers the potential for improved adventitious virus screening in vaccines and biotherapeutics. Successful implementation of such assays will require appropriate control materials to confirm assay performance and sensitivity. A common reference material containing 25 target viruses was produced and 16 laboratories were invited to process it using their preferred adventitious virus detection assay. Fifteen laboratories returned results, obtained using a wide range of wet-lab and informatics methods. Six of 25 target viruses were detected by all laboratories, with the remaining viruses detected by 4-14 laboratories. Six non-target viruses were detected by three or more laboratories. The study demonstrated that a wide range of methods are currently used for adventitious virus detection screening in biological products by deep sequencing and that they can yield significantly different results. This underscores the need for common reference materials to ensure satisfactory assay performance and enable comparisons between laboratories

Analysis of Genome Sequences from Plant Pathogenic Rhodococcus Reveals Genetic Novelties in Virulence Loci

Members of Gram-positive Actinobacteria cause economically important diseases to plants. Within the Rhodococcus genus, some members can cause growth deformities and persist as pathogens on a wide range of host plants. The current model predicts that phytopathogenic isolates require a cluster of three loci present on a linear plasmid, with the fas operon central to virulence. The Fas proteins synthesize, modify, and activate a mixture of growth regulating cytokinins, which cause a hormonal imbalance in plants, resulting in abnormal growth. We sequenced and compared the genomes of 20 isolates of Rhodococcus to gain insights into the mechanisms and evolution of virulence in these bacteria. Horizontal gene transfer was identified as critical but limited in the scale of virulence evolution, as few loci are conserved and exclusive to phytopathogenic isolates. Although the fas operon is present in most phytopathogenic isolates, it is absent from phytopathogenic isolate A21d2. Instead, this isolate has a horizontally acquired gene chimera that encodes a novel fusion protein with isopentyltransferase and phosphoribohydrolase domains, predicted to be capable of catalyzing and activating cytokinins, respectively. Cytokinin profiling of the archetypal D188 isolate revealed only one activate cytokinin type that was specifically synthesized in a fas-dependent manner. These results suggest that only the isopentenyladenine cytokinin type is synthesized and necessary for Rhodococcus phytopathogenicity, which is not consistent with the extant model stating that a mixture of cytokinins is necessary for Rhodococcus to cause leafy gall symptoms. In all, data indicate that only four horizontally acquired functions are sufficient to confer the trait of phytopathogenicity to members of the genetically diverse clade of Rhodococcus

Communications inter-règnes dans les interactions plantes-microorganismes

info:eu-repo/semantics/nonPublishe

Les microARNs: biogenèse, fonctionnement et rôles de ces nouveaux interrupteurs moléculaires

info:eu-repo/semantics/nonPublishe

Molecular bases of the Rhodococcus fascians - plant interaction :bacterial signal molecules and early plant gene responses

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Rhéostats moléculaires: les miRNAs et les siRNAs, ou comment réguler l’expression des protéines avec des petits ARNs

info:eu-repo/semantics/nonPublishe

Le Cylce Cellulaire chez les Plantes

info:eu-repo/semantics/nonPublishe