Ralstonia solanacearum is a phytopathogenic
beta-proteobacterium capable of infecting a large number of plant
species. Genoscope has sequenced the genomes of three strains with
very different host ranges. The first genome sequenced was that of a
strain with a wide host range, GMI1000. It is organized in two
replicons, a chromosome of 3.7 Mb and a megaplasmid of 2.1 Mb, with
very similar GC% (about 67%). About 10 genome equivalents were
assembled using a sequential strategy in order to limit the problems
caused by repetitive sequences. This assembly benefited from end
sequencing of BAC clones with an average insert size of 100 kb. The
resulting sequence was subjected to a finishing phase, and the final
sequence was over 99% validated by comparison with experimentally
obtained restriction enzyme fingerprints on a minimum tiling path of
BAC clones. Annotation was carried out by
Christian
Boucher's group at the Laboratoire Interactions
Plantes-Microorganismes (
access
to the annotated sequence).
The genomes of Molk2 (a strain of race 2, isolated from a banana tree)
and 1609 (a race 3 strain, isolated from potato) have genomes which
are comparable to that of GMI1000 (about 6 Mb, GC% estimated at
67%). They are being sequenced at Genoscope at depths of 6.2X and 7.3X
respectively. Three libraries are being used for each project: two
libraries of plasmid clones with average insert sizes of 3 and 10 kb,
and a library of miniBACs with an average insert size of 25 kb. After
a first assembly, finishing will only be carried out on regions of
special interest, in each genome.