Functional analyses of laccase genes from Fusarium oxysporum.

Six laccase genes, lcc1, lcc2, lcc3, lcc4, lcc5, and lcc9, of the vascular wilt fungus Fusarium oxysporum were isolated and characterized. All genes have the characteristic conserved domains for copper binding of phenol oxidase enzymes. Targeted inactivation of lcc1, lcc3, and lcc5 resulted in a significant decrease of extracellular laccase activity. Reverse transcription-polymerase chain reaction showed that lcc1, lcc2, and lcc9 were constitutively expressed in culture, whereas lcc3 and lcc5 appeared down and up-regulated, respectively, by PacC. Oxidative stress conditions and phenolic compounds altered the growth rate of the Deltalcc3 mutant compared with the wild-type. lcc1, lcc3, and lcc9 were expressed in roots and stems during the infection process. However, inactivation of lcc1, lcc3, and lcc5 had no detectable effects on virulence on tomato plants.

Role in pathogenesis of two endo-beta-1,4-xylanase genes from the vascular wilt fungus Fusarium oxysporum.

A gene, xyl4, whose predicted amino acid sequence shows significant homology with family 11 xylanases, was identified from the tomato vascular wilt fungus Fusarium oxysporum f. sp. lycopersici. Expression of xyl4 is induced on oat spelt xylan as the carbon source, subject to carbon catabolite repression and preferentially expressed at alkaline ambient pH. Transcript levels of xyl4 on an inducing carbon source are differentially regulated by the nature and concentration of the nitrogen source. As shown by RT-PCR, xyl4 is expressed by F. oxysporum during the entire cycle of infection on tomato plants. Targeted inactivation of xyl4 and of xyl3, a previously identified gene of F. oxysporum f. sp. lycopersici encoding a family 10 xylanase, had no detectable effect on virulence on tomato plants, demonstrating that both genes are not essential for pathogenicity.