A conserved fungal glycosyltransferase facilitates pathogenesis of plants by enabling hyphal growth on solid surfaces

Article Authors Metrics Comments Related Content Abstract Author summary Introduction Results and discussion Conclusions Methods Supporting information Acknowledgments References Reader Comments (0) Media Coverage (0) Figures Abstract Pathogenic fungi must extend filamentous hyphae across solid surfaces to cause diseases of plants. However, the full inventory of genes which support this is incomplete and many may be currently concealed due to their essentiality for the hyphal growth form. During a random T-DNA mutagenesis screen performed on the pleomorphic wheat (Triticum aestivum) pathogen Zymoseptoria tritici, we acquired a mutant unable to extend hyphae specifically when on solid surfaces. In contrast “yeast-like” growth, and all other growth forms, were unaffected. The inability to extend surface hyphae resulted in a complete loss of virulence on plants. The affected gene encoded a predicted type 2 glycosyltransferase (ZtGT2). Analysis of >800 genomes from taxonomically diverse fungi highlighted a generally widespread, but discontinuous, distribution of ZtGT2 orthologues, and a complete absence of any similar proteins in non-filamentous ascomycete yeasts. Deletion mutants of the ZtGT2 orthologue in the taxonomically un-related fungus Fusarium graminearum were also severely impaired in hyphal growth and non-pathogenic on wheat ears. ZtGT2 expression increased during filamentous growth and electron microscopy on deletion mutants (ΔZtGT2) suggested the protein functions to maintain the outermost surface of the fungal cell wall. Despite this, adhesion to leaf surfaces was unaffected in ΔZtGT2 mutants and global RNAseq-based gene expression profiling highlighted that surface-sensing and protein secretion was also largely unaffected. However, ΔZtGT2 mutants constitutively overexpressed several transmembrane and secreted proteins, including an important LysM-domain chitin-binding virulence effector, Zt3LysM. ZtGT2 likely functions in the synthesis of a currently unknown, potentially minor but widespread, extracellular or outer cell wall polysaccharide which plays a key role in facilitating many interactions between plants and fungi by enabling hyphal growth on solid matrices

Molecular characterization of environmental Cryptococcus neoformans isolated in Vitoria, ES, Brazil Caracterização molecular de cepas ambientais de Cryptococcus neoformans isoladas em Vitória, ES, Brasil

Cryptococcus neoformans is the major cause of fungal meningitis, a potentially lethal mycosis. Bird excreta can be considered a significant environmental reservoir of this species in urban areas, thirty-three samples of pigeon excreta were collected within the city of Vitoria, Brazil. Cryptococcus neoformans was isolated and identified using standard biochemical assays in ten samples. PCR amplification with primer M13 and orotidine monophosphate pyrophosphorylase (URA5) gene-restriction fragment length polymorphism (RFLP) analysis discerned serotypes and genotypes within this species. All isolates were serotype A (C. neoformans var. grubii) and genotype VNI. The two alternative alleles a and &#945; at the mating type locus were determined by PCR amplification and mating assays performed on V8 medium. All isolates were MAT &#945; mating type but only 50% were able to mate in vitro with the opposite mating type MAT a tester strains (JEC20, KN99a and Bt63). This study adds information on the ecology and molecular characterization of C. neoformans in the Southeast region of Brazil.<br>O "complexo Cryptococcus neoformans" é constituído por C. neoformans var. neoformans, C. neoformans var. grubii, e C. gattii. Trinta por cento de amostras de excrementos de pombos coletados dentro da cidade de Vitória, Brasil, foram positivas para Cryptococcus neoformans, espécie identificada por testes bioquímicos convencionais. Amplificação por PCR com primer M13 e análise por orotidine monophosphate pyrophosphorylase (URA5) gene-"restriction fragment length polymorphism" (RFLP) distinguiram sorotipos e genotipos dentro desta espécie. Todos os isolados ambientais foram sorotipo A (C. neoformans var. grubii) e genotipo VNI. Os dois alelos alternativos a e &#945; do locus "mating type" foram determinados por PCR e por testes de "mating" em meio V8. Todos os isolados foram "mating type" tipo MAT &#945; mas somente 50% foram capazes de conjugar in vitro com cepas MAT a, de "mating type" oposto (JEC20, KN99a e Bt63). Este estudo adiciona informações sobre a ecologia e caracterização molecular de cepas ambientais de C. neoformans, isoladas na região sudeste do Brasil

Characterization of Cryptococcus neoformans isolated from urban environmental sources in Goiânia, Goiás State, Brazil Caracterização de Cryptococcus neoformans isolados de fontes ambientais urbanas na cidade de Goiânia, estado de Goiás, Brasil

Cryptococcus neoformans is an opportunistic fungal pathogen that causes meningoencephalitis as the most frequent clinical presentation in immunocompromised patients, mainly in people infected by HIV. This fungus is an environmental encapsulated yeast, commonly found in soil enriched with avian droppings and plant material. A total of 290 samples of pigeon and the other avian droppings, soil, ornamental trees and vegetable material associated with Eucalyptus trees were collected to study environmental sources of Cryptococcus species in Goiânia, Goiás State. The determination of varieties, serotypes and the susceptibility in vitro to fluconazole, itraconazole and amphotericin B of C. neoformans isolates were performed. C. neoformans var. grubii (serotype A) was found in 20.3% (36/177) of pigeon dropping samples and in 14.3% (5/35) of samples of Eucalyptus. None of the environmental isolates of C. neoformans showed in vitro resistance to three antifungal agents. The knowledge of major route for human cryptococcal infection (inhalation of infectious particles from saprophytic sources) and a total of 60 C. neoformans isolates obtained from AIDS patients with cryptococcal meningitis between October 2001 and April 2002 justify the study of the habitats of these yeasts as probable sources of cryptococcosis in this city.<br>Cryptococcus neoformans é um fungo patogênico oportunista que causa meningoencefalite como a apresentação clínica mais importante em pacientes imunocomprometidos, principalmente, em pessoas infectadas pelo HIV. O agente é uma levedura encapsulada, comumente encontrada em solo enriquecido com excretas de aves e em resíduos de plantas. O total de 290 amostras de excretas de pombos e outras aves, de árvores ornamentais e materiais vegetais de Eucalyptus foram coletadas para estudar possíveis fontes ambientais de Cryptococcus spp, na cidade de Goiânia, Goiás. A determinação das variedades, sorotipos e suscetibilidade in vitro frente a fluconazol, itraconazol e anfotericina B dos isolados de C. neoformans foram realizadas. C. neoformans var. grubii (sorotipo A) foi a única isolada, ocorrendo em 36 (20.3%) das 177 amostras fecais de pombos e em 5 (14.3%) das 35 amostras de Eucalyptus. Nenhum dos isolados ambientais de C. neoformans mostrou resistência in vitro aos três antifúngicos avaliados. O conhecimento da principal via para infecção criptocócica humana, isto é inalação de partículas infecciosas de fontes saprofíticas e a ocorrência de 60 casos de criptococose em pacientes com AIDS, em Goiânia, entre outubro de 2001 e abril de 2002, justificam o estudo de habitats do agente como prováveis fontes de criptococose nesta cidade