Albicidin pathotoxin produced by Xanthomonas albilineans is encoded by three large PKS and NRPS genes present in a gene cluster also containing several putative modifying, regulatory, and resistance genes.

Xanthomonas albilineans, which causes leaf scald disease of sugarcane, produces a highly potent pathotoxin called albicidin. We report here sequencing and homology analysis of the major gene cluster, XALB1 (55,839 bp), and a second, smaller region, XALB2 (2,986 bp), involved in albicidin biosynthesis. XALB1 contains 20 open reading frames, including i) three large genes with a modular architecture characteristic of polyketide synthases (PKSs) and nonribosomal peptide synthases (NRPSs) and ii) several putative modifying, regulatory, and resistance genes. Sequencing and complementation studies of six albicidin-defective mutants enabled us to confirm the involvement of the three PKS and NRPS genes encoded by XALB1 in albicidin production. XALB2 contains only one gene that is required for post-translational activation of PKS and NRPS enzymes, confirming the involvement of these enzymes in albicidin biosynthesis. In silico analysis of these three PKS or NRPS enzymes allowed us to propose a model for the albicidin backbone assembly and to gain insight into the structural features of this pathotoxin. This is the first description of a complete mixed PKS-NRPS gene cluster for toxin production in the genus Xanthomonas.

The MAP kinase-encoding gene MgFus3 of the non-appressorium phytopathogen Mycosphaerella graminicola is required for penetration and in vitro pycnidia formation.

SUMMARY In eukaryotes, a family of serine/threonine protein kinases known as mitogen-activated protein kinases (MAPKs) is involved in the transduction of a variety of extracellular signals and in the regulation of growth and development. We identified a MAPK-encoding gene in Mycosphaerella graminicola strain IPO323 with high homology to the orthologous Fus3 gene of Saccharomyces cerevisiae and designated it MgFus3. Early colony development of the MgFus3 mutants during in vitro growth was similar to those of the wild-type and ectopic controls, but at the later stages of growth MgFus3 mutants did not become melanized, showed altered polarized growth and were unable to produce aerial mycelia. The MgFus3 mutants were non-pathogenic, and detailed microscopic analyses revealed that they failed to colonize the mesophyll tissue owing to the inability to penetrate stomata. Unlike the wild-type strain, MgFus3 mutants were unable to differentiate pycnidia on plant-derived media. Thus, in addition to the crucial role of MgFus3 in the regulation of penetration, it may also be involved in regulating asexual fructification. Hence, MgFus3 can be regarded as a multifunctional pathogenicity factor of M. graminicola.