Microsatellite flanking region similarities among different loci within insect species.

Although microsatellites are ubiquitous in eukaryota, the number of available markers varies strongly among taxa. This meta-analysis was conducted on 32 insect species. Sequences were obtained from two assembled whole genomes, whole genome shotgun (WGS) sequences from 10 species and screening partial genomic libraries for microsatellites from 23 species. We have demonstrated: (1) strong differences in the abundance of microsatellites among species; (2) that microsatellites within species are often grouped into families based on similarities in their flanking sequences; (3) that the proportion of microsatellites grouped into families varies strongly among taxa; and (4) that microsatellite families were significantly more often associated with transposable elements - or their remnants - than unique microsatellite sequences.

Molecular characterization of a subtilase from the vascular wilt fungus Fusarium oxysporum.

The gene prt1 was isolated from the tomato vascular wilt fungus Fusarium oxysporum f. sp. lycopersici, whose predicted amino acid sequence shows significant homology with subtilisin-like fungal proteinases. Prt1 is a single-copy gene, and its structure is highly conserved among different formae speciales of F. oxysporum. Prt1 is expressed constitutively at low levels during growth on different carbon and nitrogen sources and strongly induced in medium containing collagen and glucose. As shown by reverse transcription-polymerase chain reaction and fluorescence microscopy of F. oxysporum strains carrying a prt1-promoter-green fluorescent protein fusion, prt1 is expressed at low levels during the entire cycle of infection on tomato plants. F. oxysporum strains transformed with an expression vector containing the prt1 coding region fused to the inducible endopolygalacturonase pg1 gene promoter and grown under promoter-inducing conditions secreted high levels of extracellular subtilase activity that resolved into a single peak of pI 4.0 upon isoelectric focusing. The active fraction produced two clearing bands of 29 and 32 kDa in sodium dodecyl sulfate gels containing gelatin. Targeted inactivation of prt1 in F. oxysporum f. sp. lycopersici had no detectable effect on mycelial growth, sporulation, and pathogenicity on tomato plants.

A MAP kinase of the vascular wilt fungus Fusarium oxysporum is essential for root penetration and pathogenesis.

The soil-borne vascular wilt fungus Fusarium oxysporum infects a wide variety of plant species by directly penetrating roots, invading the cortex and colonizing the vascular tissue. We have identified fmk1, encoding a mitogen-activated protein kinase (MAPK) of F. oxysporum that belongs to the yeast and fungal extracellular signal-regulated kinase (YERK1) subfamily. Targeted mutants of F. oxysporum f. sp. lycopersici carrying an inactivated copy of fmk1 have lost pathogenicity on tomato plants but show normal vegetative growth and conidiation in culture. Colonies of the fmk1 mutants are easily wettable, and hyphae are impaired in breaching the liquid-air interface, suggesting defects in surface hydrophobicity. Fmk1 mutants also show reduced invasive growth on tomato fruit tissue and drastically reduced transcript levels of pl1 encoding the cell wall-degrading enzyme pectate lyase. Conidia of the mutants germinating in the tomato rhizosphere fail to differentiate penetration hyphae, resulting in greatly impaired root attachment. The orthologous MAPK gene Pmk1 from the rice leaf pathogen Magnaporthe grisea complements invasive growth and partially restores surface hydrophobicity, root attachment and pathogenicity in an fmk1 mutant. These results demonstrate that FMK1 controls several key steps in the pathogenesis of F. oxysporum and suggest a fundamentally conserved role for the corresponding MAPK pathway in soil-borne and foliar plant pathogens.

Role of cell wall-degrading enzymes in pathogenicity of Fusarium oxysporum.

Fusarium oxysporum invades its host plants through the roots and colonizes the vascular system. It produces a great variety of cell-wall degrading enzymes (CWDE), such as cellulases, xylanases, pectinases and proteases. Our group has purified and characterized an endopolygalacturonase (PG1), two exopolygalacturonases (PG2 and PG3), an endoxylanase (XYL1) and an endo pectatelyase (PL1). We have isolated the following CWDE-encoding genes: pg1, pgx4, pg5, xyl2, xyl3, prt1 and pl1. Gene expression in different culture conditions has been determined by Northern analysis. The occurrence of these genes in different formae speciales has been analyzed by Southern analysis and PCR. All these genes are expressed during different stages of the interaction with the host plant indicating a possible role in pathogenesis. At present, targeted gene disruption is being carried out, in order to determine the role of each gene in the pathogenicity process.

Allozyme variation in Parnassius mnemosyne (L.) (Lepidoptera) populations in North-East Hungary: variation within a subspecies group.

Allozyme polymorphism was studied in 11 Parnassius mnemosyne (Linnaeus, 1758) populations in North-East Hungary. Significant departures from Hardy-Weinberg equilibrium were observed in several cases due to heterozygote deficiency. Genetic variability did not display geographical pattern; the level of genetic differentiation was similar between adjacent populations and between populations originating from different geographical regions. Even a completely isolated population was not differentiated markedly from the others. Thus, genetic drift can be expected as the main evolutionary force acting in the populations.

Enzyme polymorphism in Drosophila melanogaster populations collected in two different habitats in Hungary.

The level of enzyme polymorphism was compared in ten Drosophila melanogaster populations collected in farmyards and distilleries in two regions of Hungary. The total genetic diversity was partitioned into between- and within-population components at each investigated locus using Wright's F-statistics. Population differentiation was studied in two different ways. Genetic distances between pairs of populations were calculated and a hierarchical analysis of gene diversity was performed. Based on the F values gene flow was estimated among the populations at different levels of the hierarchy. The results indicated that our 'farmyard populations' collected within a region could be considered as parallel samples from a panmictic population rather than samples of distinct populations. In distilleries, the flies might be influenced by two different evolutionary forces: (i) selection due to the extremely high concentration of ethanol in the fermenting mash and (ii) genetic drift due to the combination of repeated founder effects and fluctuating population size. Our results suggested that 'distillery populations' could not be regarded as real populations either. They could be considered as peculiar cases: founder individuals taken from the total population (region) established special populations which survived in the distilleries for many generations. Thus the dominating force acting on the 'distillery populations' was genetic drift.