Ralstonia solanacearum is a devastating, soil-borne plant pathogen with a global distribution and an unusually wide host range. It is a model system for the dissection of molecular determinants governing pathogenicity. We present here the complete genome sequence and its analysis of strain GMI1000. The 5.8-megabase (Mb) genome is organized into two replicons: a 3.7-Mb chromosome and a 2.1-Mb megaplasmid. Both replicons have a mosaic structure providing evidence for the acquisition of genes through horizontal gene transfer. Regions containing genetically mobile elements associated with the percentage of G+C bias may have an important function in genome evolution. The genome encodes many proteins potentially associated with a role in pathogenicity. In particular, many putative attachment factors were identified. The complete repertoire of type III secreted effector proteins can be studied. Over 40 candidates were identified. Comparison with other genomes suggests that bacterial plant pathogens and animal pathogens harbour distinct arrays of specialized type III-dependent effectors.
The advent of fully sequenced genomes opens the ground for the reconstruction of metabolic pathways on the basis of the identification of enzyme-coding genes. Here we describe PRIAM, a method for automated enzyme detection in a fully sequenced genome, based on the classification of enzymes in the ENZYME database. PRIAM relies on sets of position-specific scoring matrices ('profiles') automatically tailored for each ENZYME entry. Automatically generated logical rules define which of these profiles is required in order to infer the presence of the corresponding enzyme in an organism. As an example, PRIAM was applied to identify potential metabolic pathways from the complete genome of the nitrogen-fixing bacterium Sinorhizobium meliloti. The results of this automated method were compared with the original genome annotation and visualised on KEGG graphs in order to facilitate the interpretation of metabolic pathways and to highlight potentially missing enzymes.
Studies of the interaction between Arabidopsis thaliana and the necrotrophic fungal pathogen Sclerotinia sclerotiorum have been hampered by the extreme susceptibility of this model plant to the fungus. In addition, analyses of the plant defense response suggested the implication of a complex interplay of hormonal and signaling pathways. To get a deeper insight into this host-pathogen interaction, we first analyzed the natural variation in Arabidopsis for resistance to S. sclerotiorum. The results revealed a large variation of resistance and susceptibility in Arabidopsis, with some ecotypes, such as Ws-4, Col-0, and Rbz-1, being strongly resistant, and others, such as Shahdara, Ita-0, and Cvi-0, exhibiting an extreme susceptibility. The role of different signaling pathways in resistance was then determined by assessing the symptoms of mutants affected in the perception, production, or transduction of hormonal signals after inoculation with S. sclerotiorum. This analysis led to the conclusions that i) signaling of inducible defenses is predominantly mediated by jasmonic acid and abscisic acid, influenced by ethylene, and independent of salicylic acid; and ii) nitric oxide (NO) and reactive oxygen species are important signals required for plant resistance to S. sclerotiorum. Defense gene expression analysis supported the specific role of NO in defense activation.
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