Project description:The actinobacteria Frankia alni is able to induce the formation of nodules on the root of a large spectrum of actinorhizal plants, where it converts dinitrogen to ammonia in exchange for plant photosynthates. In the present study, transcriptional analyses were performed on nitrogen-replete free-living cells and on Alnus glutinosa nodule bacteria, using whole genome microarrays. Distribution of nodule-induced genes on the genome was found to be mostly over regions with high synteny between three Frankia genomes, while nodule-repressed genes, which were mostly hypothetical and not conserved, were spread around the genome. Genes known to be related to symbiosis were highly induced: nif (nitrogenase), hup2 (hydrogenase uptake), suf (sulfur-iron cluster) and shc (hopanoids synthesis). The expression of genes involved in ammonium assimilation and transport was strongly modified suggesting that bacteria ammonium assimilation was limited. Genes involved in particular in transcriptional regulation, signalling processes, protein drug export, protein secretion, lipopolysaccharide and peptidoglycan biosynthesis that may play a role in symbiosis were also identified. We showed that this nodule transcriptome of Frankia was highly similar among phylogenetically distant plant families. To address gene expression changes of Frankia alni ACN in the symbiotic state, we compared transcript levels between young nodules formed on 4 species of trees (Alnus glutinosa, Alnus nepalensis, Myrica gale and Myrica rubra) and free-living cells grown in nitrogen-replete minimal medium. For A. glutinosa nodule and free-living cells, two sets of experiments (A and B) were performed in two different laboratories. Three biological replicates were preformed for each condition.
Project description:This work introduces a novel online Headspace-Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry-based untargeted metabolomics approach, suggested as an alternative tool to study the environmental fate of volatile xenometabolites in emerging complex biopesticides, e.g. the Myrica gale methanolic extract herbicide containing several unknown metabolites. A 'living' microcosm sample was designed for non-destructive analysis by a 35-min HS-SPME automated extraction and a 36-min GC-MS run. A 38-day kinetics study was then applied on two groups of soil samples: control and spiked. Statistical tools were used for the comparative kinetics study. The Principal Component Analysis revealed and explained the evolution and the dissipation of the herbicide volatile xenometabolome over time. The time-series Heatmap and Multivariate Empirical Bayes Analysis of Variance allowed the prioritization of 101 relevant compounds including 22 degradation by-products. Out of them, 96 xenometabolites were putatively identified. They included 63 compounds that are identified as herbicide components for the first time. The Orthogonal Projections to Latent Structures Discriminant Analysis and its Cross-Validation test were used to assess the total dissipation of the herbicide volatile residues and method detection limit. The reproducibility of the method was also assessed. The highest inter-samples (n = 3) Peak Area RSD was 7.75 %. The highest inter-samples (n = 3) and inter-days (n = 8) Retention Time SD were 0.43 sec and 3.44 sec, respectively. The work presents a green, non-laborious and high-throughput approach. It required a small number of environmental samples (6 microcosms) that were analyzed 8 times and were not destroyed during the study.