The differential virulence of Fusarium strains causing corneal infections and plant diseases is associated with accessory chromosome composition.

Fusarium oxysporum is a cross-kingdom pathogen. While some strains cause disseminated fusariosis and blinding corneal infections in humans, others are responsible for devastating vascular wilt diseases in plants. To better understand the distinct adaptations of F. oxysporum to animal or plant hosts, we conducted a comparative phenotypic and genetic analysis of two strains: MRL8996 (isolated from a keratitis patient) and Fol4287 (isolated from a wilted tomato [Solanum lycopersicum]). In vivo infection of mouse corneas and tomato plants revealed that, while both strains cause symptoms in both hosts, MRL8996 caused more severe corneal ulceration and perforation in mice, whereas Fol4287 induced more pronounced wilting symptoms in tomato. In vitro assays using abiotic stress treatments revealed that the human pathogen MRL8996 was better adapted to elevated temperatures, whereas the plant pathogen Fol4287 was more tolerant of osmotic and cell wall stresses. Both strains displayed broad resistance to antifungal treatment, with MRL8996 exhibiting the paradoxical effect of increased tolerance to higher concentrations of the antifungal caspofungin. We identified a set of accessory chromosomes (ACs) and protein-encoding genes with distinct transposon profiles and functions, respectively, between MRL8996 and Fol4287. Interestingly, ACs from both genomes also encode proteins with shared functions, such as chromatin remodeling and post-translational protein modifications. Our phenotypic assays and comparative genomics analyses lay the foundation for future studies correlating genotype with phenotype and for developing targeted antifungals for agricultural and clinical uses.

The study of iron- and copper-binding proteome of Fusarium oxysporum and its effector candidates.

Fusarium oxysporum f.sp. lycopersici is a phytopathogen which causes vascular wilt disease in tomato plants. The survival tactics of both pathogens and hosts depend on intricate interactions between host plants and pathogenic microbes. Iron-binding proteins (IBPs) and copper-binding proteins (CBPs) play a crucial role in these interactions by participating in enzyme reactions, virulence, metabolism, and transport processes. We employed high-throughput computational tools at the sequence and structural levels to investigate the IBPs and CBPs of F. oxysporum. A total of 124 IBPs and 37 CBPs were identified in the proteome of Fusarium. The ranking of amino acids based on their affinity for binding with iron is Glu > His> Asp > Asn > Cys, and for copper is His > Asp > Cys respectively. The functional annotation, determination of subcellular localization, and Gene Ontology analysis of these putative IBPs and CBPs have unveiled their potential involvement in a diverse array of cellular and biological processes. Three iron-binding glycosyl hydrolase family proteins, along with four CBPs with carbohydrate-binding domains, have been identified as potential effector candidates. These proteins are distinct from the host Solanum lycopersicum proteome. Moreover, they are known to be located extracellularly and function as enzymes that degrade the host cell wall during pathogen-host interactions. The insights gained from this report on the role of metal ions in plant-pathogen interactions can help develop a better understanding of their fundamental biology and control vascular wilt disease in tomato plants.

First report of Bretziella fagacearum infecting chestnut in Michigan.

In the summer of 2021, a 20-year-old 'Colossal' (Castanea sativa × C. crenata hybrid) tree in a commercial chestnut orchard in northwest Michigan suddenly declined. Until 2023, an additional 26 adjacent trees declined, suggesting the occurrence of root-graft transmission of the pathogen. The initial wilting of leaves progressed to complete tree death in about 10 days. Symptoms included wilting, and bronzing, followed by tanning starting at leaf apex and margins, with significant defoliation. Sometimes black-to-brown streaks of discoloration appear in the sapwood, with no signs of mycelial mat production on dead trees. Branches from symptomatic trees in two different areas of the orchard were submitted to Plant and Pest Diagnostics at Michigan State University. Bretziella fagacearum (Bretz) Z.W. de Beer, Marinc., T.A. Duong & M.J. Wingf. was detected in both samples using nested PCR (Wu et al. 2011) and qPCR (Bourgault et al. 2022). The products of the nested PCR were sequenced (GenBank accession nos. OR522695-OR522696) and BLASTn search results showed 100% identity to an ex-type strain of B. fagacearum (MH865866). Surface-sterilized discolored sapwood chips were plated on acidified potato dextrose agar (aPDA). Bretziella fagacearum was consistently recovered; colony and endoconidia morphology aligned with the description of the pathogen (De Beer et al. 2017). A pure culture (BF277) was obtained for inoculation experiments. To confirm pathogenicity, 10 'Colossal' chestnut seedlings (average stem diameter of 9 mm) were inoculated in the greenhouse with a 14-day old culture of BF277. Using a conical drill bit, two 0.4 mm diameter holes were drilled, one was 5 cm above the soil line at a 45° angle and the other was on the opposite side of the stem at least 10 cm above the soil line. A 50-µl conidial suspension (1 × 107 conidia per ml) was applied and the holes were sealed with Parafilm. Five 'Colossal' seedlings were inoculated with sterile water. Leaf epinasty with bent petioles was observed 14 days later. Leaf wilting and necrosis similar to natural infection in the orchard were observed at 24 and 34 days after inoculation, respectively. Water-inoculated control plants showed no symptoms. Bretziella fagacearum was reisolated from symptomatic plants by surface sterilizing leaf petioles with 75% ethanol (30 s), followed by 10% (v/v) bleach (1 min), and two rinses with sterile deionized water (>1 min). Petiole pieces (~1 cm) were plated on aPDA. The pathogen was reisolated from six symptomatic plants and detected using qPCR in the remaining four seedlings. Bretziella fagacearum was not detected in control plants. The identity of the recovered fungus was confirmed following the amplification of the internal transcribed spacer (ITS) from extracted genomic DNA, as described in Chahal et al. 2022. The resulting PCR product was sequenced and assembled into a consensus sequence using Geneious Prime. The consensus sequence (accession no. OR515809) revealed 100% identity to the ex-type of B. fagacearum (KU042044). This is the first record of B. fagacearum infecting chestnut trees in Michigan. Previously, B. fagacearum has been reported infecting Chinese chestnut (C. mollissima) in Missouri (Bretz and Long, 1950). Oak wilt is widely distributed in Michigan and is the predominant disease afflicting red oaks in the Midwestern U.S. Consequently, constant vigilance and monitoring are essential in chestnut orchards to promptly detect and effectively manage potential infections.

Morphophysiological characterization of Ceratocystis fimbriata isolates from yerba mate / Caracterização morfofisiológica de isolados de Ceratocystis fimbriata de erva-mate

ABSTRACT: This study aimed to morphologically characterize the isolates of Ceratocystis fimbriata from yerba mate and to evaluate the effect of culture medium and temperature on mycelial growth and sporulation of C. fimbriata. For the morphological characterization of the 11 monosporic isolates of the fungus, slides were prepared to determine the dimensions of the sexual and asexual structures of the fungus. Four experiments were conducted to evaluate the mycelial growth and to evaluate the sporulation of C. fimbriata in different culture mediums and temperatures. The isolates of C. fimbriata from yerba mate showed perithecia with brown to black necks, divergent ostiolar hyphae, hatshaped hyaline ascospores, single-celled, cylindrical endoconidia, and globular to ovoid aleurioconidia. PDA and V8-agar media showed the highest mycelial growth. The average optimum temperature for mycelial growth and sporulation of isolates of C. fimbriata of yerba mate were 22.5 and 22.4 ºC, respectively.

RESUMO: Este estudo teve como objetivo caracterizar morfologicamente os isolados de Ceratocystis fimbriata e avaliar o efeito do meio de cultura e da temperatura no crescimento micelial e na esporulação de C. fimbriata. Para a caracterização morfológica dos 11 isolados monospóricos do fungo foram preparadas lâminas para determinar as dimensões das estruturas sexuadas e assexuadas do fungo. Quatro experimentos foram conduzidos para avaliar o crescimento micelial e esporulação of C. fimbriata em diferentes meios de culturae temperaturas. Os isolados de C. fimbriata de erva-mate apresentaram peritécios com pescoço de marrom a preto, hifa ostiolar divergente, ascósporos hialinos em formato de chapéu, endoconídios unicelulares, cilíndricos, e aleuroconídios com formato globoso a ovoide. Os meios de cultura PDA e V8-ágar apresentaram os maiores crescimentos miceliais. A temperatura ótima média para crescimento micelial e esporulação dos isolados de C. fimbriata de erva-mate foram de 22,5 e 22,4 ºC, respectivamente.

Hongos fitopatógenos asociados a la enfermedad de muerte regresiva y pudrición del fuste de Pachaco (Schizolobium parahybum) en el trópico húmedo ecuatoriano / Fungi pathogens associated disease dieback and stem rot of Pachaco (Schizolobium parahybum) in the Ecuadorian humed tropics

La enfermedad de muerte regresiva y pudrición del fuste de Pachaco (Schizolobium parahybum Vell Blake) fue detectada inicialmente 1988, matando árboles jóvenes en la zona central del Trópico Húmedo Ecuatoriano, desde entonces, cada año, miles de árboles mueren por esta causa. En el año 2002 se realizó la presente investigación que abarcó tres rodales de distintas edades, ubicados en provincias diferentes, planteándose los siguientes objetivos: describir la sintomatología de la enfermedad, su incidencia en las zonas de estudio e identificar los microorganismos asociados a árboles de S. parahybum enfermos. La enfermedad mostró una sintomatología compleja, empezando con una marchitez vascular y perdida de coloración verde oscura de las hojas hasta tornarse cloróticas. Las heridas no cicatrizadas mostraron una pudrición circular que afecta la corteza y el xilema, emanando fluidos de color amarillo claro a café oscuro, con fuerte olor a materia orgánica en descomposición. En la provincia de Los Ríos (rodal mayor a 25 años) se encontró la mayor incidencia de la enfermedad (55 por ciento), seguido de las provincias Esmeraldas (rodal de 8 años) y Santo Domingo de Los Tsáchilas (rodal de 4 años) con 29 y 24 por ciento, respectivamente. De árboles enfermos se aislaron e identificaron cinco hongos, tres a nivel de género: Macrophoma sp., Fusarium sp. y Graphium sp. y dos a nivel de especies: Ceratocystis paradoxa (Dade) y C. moniliformis (Hedgc.). Las pruebas de patogenicidad demostraron que C. paradoxa, Macrophoma sp. y C. moniliformis provocaron los mayores valores de volumen aparente de necrosis con 5.40, 3.55 y 3.48 cm3respectivamente.

Dieback disease and stem rot of Pachaco (Schizolobium parahybum Vell Blake) was detected initially in 1988, killing young trees in the Ecuadorian Humed Tropics, since then, thousands of trees are dying annually. The present investigation was made in 2002, where were included three rodales of different ages, located in different provinces. The aim for this research was describe the symptoms its incidence in the zones of study and identifying the associated microorganisms affecting S. parahybum. The disease showed complex symtoms, beginning with a wither of dark, green coloration of the leaves until becoming clorotics. The healed wounds did not show a circular root that affects the crust and xylem, of where emanates fluid of clear yellow color to dark coffee, with strong smell to organic matter in decomposition. In Los Ríos province (rodal more than 25 years old) the greater incidence of the disease was 55 percent, followed by he provinces of Esmeraldas (rodal of 8 years old) and Santo Domingo de Los Tsáchilas (rodal of 4 years old) with 29 and 24 percent respectively. Of ill trees five fungi isolated themselves and identified, three at sort level: Macrophoma sp., Fusarium sp. and Graphium sp. and two at level of species: Ceratocystis paradoxa (Dade) and C. Moniliformis (Hedgc.). The pathogenicity tests demonstrated that C. paradoxa, Macrophoma sp. and C. moniliformis caused the higher values of apparent volume of necrosis with 5.40, 3.55 and 3.48 cm3 respectively.