Horizontal transfer of genetic elements in the black Aspergilli

The thesis deals with the horizontal transfer of genetic elements in the black Aspergilli . The black Aspergilli form a complex group of asexual species. All share a characteristic black conidiospore color and the ability to efficiently degrade tannin. Selective isolation of all different black Aspergillus types is possible on media with 20% tannin. Tannins can form complexes with proteins, that are difficult to mineralize. Therefore, the strains may have a special niche in the control of the natural nitrogen cycle. Black Aspergilli occur worldwide and especially in warmer regions at high densities. The spores have an efficient aerial distribution, which produces a well-mixed sporebank in soil throughout the world.Under laboratory conditions isogenic lines are capable, after hyphal fusions, to form a heteroplasmic heterokaryon and (transient) diploids. This so-called parasexual cycle can result in recombination via reassortment of chromosomes, mitotic crossing-over and/or exchange of cytoplasmic genetic elements. Most of the natural isolates are heterokaryon incompatible with one another and unable to form a stable heterokaryon. About the exact mechanism of the heterokaryon incompatibility reactions in the black Aspergilli little is known. Confrontations between heterokaryon self-incompatible strains suggest that prefusion genes are involved. The fact that protoplast fusions are partly able to overcome incompatibility reactions suggests that also fusion and postfusion genes are involved.One of the cytoplasmic candidates for horizontal transfer is the mitochondrion. Different mitochondrial haplotypes can be distinguished, corresponding with different black Aspergillus types . No horizontal transfer or recombination of mitochondria was observed in our natural isolates, though in protoplast fusions mitochondria can recombine. In nature new mitochondrial types may result from mutations.Most of the transfer experiments in these thesis were done with cytoplamsic dsRNA mycoviruses. In nature 10% of the population is infected with a variety of different dsRNA fragments of different viral origins. These mycoviruses can cause serious reductions in their host's fitness on traits as spore production and growth rate. Population genetic models predict that deleterious elements should disappear from a population, unless they have an extra way of transfer than just vertical transmission to offspring. Interspecies transfer of mycoviruses with species like Fusarium poae was in our experiments less difficult to achieve (and thus perhaps more likely in nature) than intraspecies transfer between different black Aspergillus types.In a diploid both interchromosomal and intrachromosomal mitotic recombination could take place. However, molecular data suggests that there is little (para)sexual recombination in the black Aspergillus population, in contrast to other presumably asexual fungi tested so far. Recently transposable elements have been found in black Aspergillus strains. These do seem to have transposed between different, quite unrelated strains. Circular intermediates of these transposable elements may also transfer little parts of genomic DNA, which may lead to some recombination. The size of the genetic elements may influence the chance on horizontal transfer during cell contact: no detectable transfer of mitochondria, very little of mycoviruses and some transfer of transposable elements.</p

Calendar 2012

Background Fusarium species are among the most common fungi present in the environment and some species have emerged as major opportunistic fungal infection in human. However, in immunocompromised hosts they can be virulent pathogens and can cause death. The pathogenesis of this infection relies on three factors: colonization, tissue damage, and immunosuppression. A novel Fusarium species is reported for the first time from keratitis in an agriculture worker who acquired the infection from plant material of maize. Maize plants are the natural host of this fungus where it causes stalk rot and seeding malformation under temperate and humid climatic conditions. The clinical manifestation, microbiological morphology, physiological features and molecular data are described.MethodsDiagnosis was established by using polymerase chain reaction of fungal DNA followed by sequencing portions of translation elongation factor 1 alpha (TEF1 ¿) and beta-tubulin (BT2) genes. Susceptibility profiles of this fungus were evaluated using CLSI broth microdilution method.ResultsThe analyses of these two genes sequences support a novel opportunist with the designation Fusarium temperatum. Phylogenetic analyses showed that the reported clinical isolate was nested within the Fusarium fujikuroi species complex. Antifungal susceptibility testing demonstrated that the fungus had low MICs of micafungin (0.031 ¿g/ml), posaconazole (0.25 ¿g/ml) and amphotericin B (0.5 ¿g/ml).ConclusionThe present case extends the significance of the genus Fusarium as agents of keratitis and underscores the utility of molecular verification of these emerging fungi in the human host

Efficient degradation of tannic acid by black Aspergillus species

A set of aspergillus strains from culture collections and wild-type black aspergilli isolated on non-selective media were used to validate the use of media with 20 % tannic acid for exclusive and complete selection of the black aspergilli. The 20% tannic acid medium proved useful for both quantitative and qualitative selection of all different black aspergilli, including all recognized species: A. carbonarius, A. japonicus, A. aculeatus, A foetidus, A. heteromorphus, A. niger, A. tubingensis and A. brasiliensis haplotypes. Even higher concentrations of tannic acid can be utilized by the black aspergilli suggesting a very efficient tannic acid-degrading system. Colour mutants show that the characteristic ability to grow on high tannic acid concentrations is not causally linked to the other typical feature of these aspergilli, i.e. the formation of brown-black pigments. Sequence analysis of the A. niger genome using the A. orvzae tannase gene yielded eleven tannase-like genes, far more than in related species. Therefore, a unique ecological niche in the degradation of tannic acid and connected nitrogen release seems to be reserved for these black-spored cosmopolitans

Diversity of Mobile Genetic Elements in the Mitogenomes of Closely Related Fusarium culmorum and F. graminearum sensu stricto Strains and Its Implication for Diagnostic Purposes

Much of the mitogenome variation observed in fungal lineages seems driven by mobile genetic elements (MGEs), which have invaded their genomes throughout evolution. The variation in the distribution and nucleotide diversity of these elements appears to be the main distinction between different fungal taxa, making them promising candidates for diagnostic purposes. Fungi of the genus Fusarium display a high variation in MGE content, from MGE-poor (Fusarium oxysporum and Fusarium fujikuroi species complex) to MGE-rich mitogenomes found in the important cereal pathogens F. culmorum and F. graminearum sensu stricto. In this study, we investigated the MGE variation in these latter two species by mitogenome analysis of geographically diverse strains. In addition, a smaller set of F. cerealis and F. pseudograminearum strains was included for comparison. Forty-seven introns harboring from 0 to 3 endonucleases (HEGs) were identified in the standard set of mitochondrial protein-coding genes. Most of them belonged to the group I intron family and harbored either LAGLIDADG or GIY-YIG HEGs. Among a total of 53 HEGs, 27 were shared by all fungal strains. Most of the optional HEGs were irregularly distributed among fungal strains/species indicating ancestral mosaicism in MGEs. However, among optional MGEs, one exhibited species-specific conservation in F. culmorum. While in F. graminearum s.s. MGE patterns in cox3 and in the intergenic spacer between cox2 and nad4L may facilitate the identification of this species. Thus, our results demonstrate distinctive traits of mitogenomes for diagnostic purposes of Fusaria

Diversity of mobile genetic elements in the mitogenome of closely related Fusarium culmorum and F. graminearum sensu stricto strains ans its implication for diagnostic purposes

Much of the mitogenome variation observed in fungal lineages seems driven by mobile genetic elements (MGEs), which have invaded their genomes throughout evolution. The variation in the distribution and nucleotide diversity of these elements appears to be the main distinction between different fungal taxa, making them promising candidates for diagnostic purposes. Fungi of the genus Fusarium display a high variation in MGE content, from MGE-poor (F. oxysporum and Fusarium fujikuroi species complex) to MGE-rich mitogenomes found in the important cereal pathogens F. culmorum and F. graminearum sensu stricto. In this study, we investigated the MGE variation in these latter two species by mitogenome analysis of geographically diverse strains. In addition, a smaller set of F. cerealis and F. pseudograminearum strains was included for comparison. Forty-seven introns harboring from 0 to 3 endonucleases (HEGs) were identified in the standard set of mitochondrial protein-coding genes. Most of them belonged to the group I intron family and harbored either LAGLIDADG or GIY-YIG HEGs. Among a total of 53 HEGs, 27 were shared by all fungal strains. Most of the optional HEGs were irregularly distributed among fungal strains/species indicating ancestral mosaicism in MGEs. However, among optional MGEs, one exhibited species-specific conservation in F. culmorum. While in F. graminearum s.s. MGE patterns in cox3 and in the intergenic spacer between cox2 and nad4L may facilitate the identification of this species. Thus, our results demonstrate distinctive traits of mitogenomes for diagnostic purposes of Fusaria.Fil: Kulik, Tomasz. Department Of Botany And Nature Protection, University; PoloniaFil: Brankovics, Balazs. Wageningen Plant Research, Wageningen University; Países BajosFil: Van Diepeningen, Anne D.. Waneningen Plant Research; Países BajosFil: Bilska, Katarzyna. Department Of Botany And Nature Protection, University; PoloniaFil: Zelechowski, Maciej. Department Of Botany And Nature Protection, University; PoloniaFil: Myszczyński, Kamil. Department Of Botany And Nature Protection, University; PoloniaFil: Molcan, Tomasz. Faculty Of Biology And Biotechnology, University; PoloniaFil: Stakheev. Alexander. Institute Of Bioorganic Chemistry (ras); RusiaFil: Stenglein, Sebastian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnolológico Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología. Laboratorio de Biología Funcional y Biotecnología; ArgentinaFil: Beyer, Marco. Luxembourg Institute Of Science And Technology; LuxemburgoFil: Pasquali, Matias. Faculty Of Agricultural And Food Sciences; ItaliaFil: Sawicki, Jakub. Department Of Botany And Nature Protection, University; PoloniaFil: Baturo Cieśniewska, Anna. Baturo-cie&#347;niewska; Poloni

On the isoperimetric problem for the Laplacian with Robin and Wentzell boundary conditions

Doctor of PhilosophyWe consider the problem of minimising the eigenvalues of the Laplacian with Robin boundary conditions $\frac{\partial u}{\partial \nu} + \alpha u = 0$ and generalised Wentzell boundary conditions $\Delta u + \beta \frac{\partial u}{\partial \nu} + \gamma u = 0$ with respect to the domain $\Omega \subset \mathbb R^N$ on which the problem is defined. For the Robin problem, when $\alpha > 0$ we extend the Faber-Krahn inequality of Daners [Math. Ann. 335 (2006), 767--785], which states that the ball minimises the first eigenvalue, to prove that the minimiser is unique amongst domains of class $C^2$. The method of proof uses a functional of the level sets to estimate the first eigenvalue from below, together with a rearrangement of the ball's eigenfunction onto the domain $\Omega$ and the usual isoperimetric inequality. We then prove that the second eigenvalue attains its minimum only on the disjoint union of two equal balls, and set the proof up so it works for the Robin $p$-Laplacian. For the higher eigenvalues, we show that it is in general impossible for a minimiser to exist independently of $\alpha > 0$. When $\alpha < 0$, we prove that every eigenvalue behaves like $-\alpha^2$ as $\alpha \to -\infty$, provided only that $\Omega$ is bounded with $C^1$ boundary. This generalises a result of Lou and Zhu [Pacific J. Math. 214 (2004), 323--334] for the first eigenvalue. For the Wentzell problem, we (re-)prove general operator properties, including for the less-studied case $\beta 0$ establish a type of equivalence property between the Wentzell and Robin minimisers for all eigenvalues. This yields a minimiser of the second Wentzell eigenvalue. We also prove a Cheeger-type inequality for the first eigenvalue in this case

Eighty Years of Mycopathologia: A Retrospective Analysis of Progress Made in Understanding Human and Animal Fungal Pathogens

Mycopathologia was founded in 1938 to \u27diffuse the understanding of fungal diseases in man and animals among mycologists.\u27 This was an important mission considering that pathogenic fungi for humans and animals represent a tiny minority of the estimated 1.5-5 million fungal inhabitants on Earth. These pathogens have diverged from the usual saprotrophic lifestyles of most fungi to colonize and infect humans and animals. Medical and veterinary mycology is the subdiscipline of microbiology that dwells into the mysteries of parasitic, fungal lifestyles. Among the oldest continuing scientific publications on the subject, Mycopathologia had its share of \u27classic papers\u27 since the first issue was published in 1938. An analysis of the eight decades of notable contributions reveals many facets of host-pathogen interactions among 183 volumes comprising about 6885 articles. We have analyzed the impact and relevance of this body of work using a combination of citation tools (Google Scholar and Scopus) since no single citation metric gives an inclusive perspective. Among the highly cited Mycopathologia publications, those on experimental mycology accounted for the major part of the articles (36%), followed by diagnostic mycology (16%), ecology and epidemiology (15%), clinical mycology (14%), taxonomy and classification (10%), and veterinary mycology (9%). The first classic publication, collecting nearly 200 citations, appeared in 1957, while two articles published in 2010 received nearly 150 citations each, which is notable for a journal covering a highly specialized field of study. An empirical analysis of the publication trends suggests continuing interests in novel diagnostics, fungal pathogenesis, review of clinical diseases especially with relevance to the laboratory scientists, taxonomy and classification of fungal pathogens, fungal infections and carriage in pets and wildlife, and changing ecology and epidemiology of fungal diseases around the globe. We anticipate that emerging and re-emerging fungal pathogens will continue to cause significant health burden in the coming decades. It remains vital that scientists and physicians continue to collaborate by learning each other\u27s language for the study of fungal diseases, and Mycopathologia will strive to be their partner in this increasingly important endeavor to its 100th anniversary in 2038 and beyond

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