Françoise Munaut

A European Database of Fusarium graminearum and F. culmorum Trichothecene Genotypes

Fusarium species, particularly Fusarium graminearum and F. culmorum, are the main cause of trichothecene type B contamination in cereals. Data on the distribution of Fusarium trichothecene genotypes in cereals in Europe are scattered in time and space. Furthermore, a common core set of related variables (sampling method, host cultivar, previous crop, etc.) that would allow more effective analysis of factors influencing the spatial and temporal population distribution, is lacking. Consequently, based on the available data, it is difficult to identify factors influencing chemotype distribution and spread at the European level. Here we describe the results of a collaborative integrated work which aims (1) to characterize the trichothecene genotypes of strains from three Fusarium species, collected over the period 2000–2013 and (2) to enhance the standardization of epidemiological data collection. Information on host plant, country of origin, sampling location, year of sampling and previous crop of 1147 F. graminearum, 479 F. culmorum, and 3 F. cortaderiae strains obtained from 17 European countries was compiled and a map of trichothecene type B genotype distribution was plotted for each species. All information on the strains was collected in a freely accessible and updatable database (www.catalogueeu.luxmcc.lu), which will serve as a starting point for epidemiological analysis of potential spatial and temporal trichothecene genotype shifts in Europe. The analysis of the currently available European dataset showed that in F. graminearum, the predominant genotype was 15-acetyldeoxynivalenol (15-ADON) (82.9%), followed by 3-acetyldeoxynivalenol (3-ADON) (13.6%), and nivalenol (NIV) (3.5%). In F. culmorum, the prevalent genotype was 3-ADON (59.9%), while the NIV genotype accounted for the remaining 40.1%. Both, geographical and temporal patterns of trichothecene genotypes distribution were identified.

One fungus, one name

In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice.

One fungus, one name: defining the genus Fusarium in a scientifically robust way that preserves longstanding use.

In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice.

Fusarium temperatum sp. nov. from maize, an emergent species closely related to Fusarium subglutinans.

A large number of Fusarium isolates closely related to F. subglutinans were collected from maize in Belgium. We used a robust polyphasic approach to describe a new biological species, Fusarium temperatum, within the Gibberella fujikuroi species complex. F. temperatum can be distinguished from F. subglutinans and from other Fusarium species within the Gibberella fujikuroi species complex with AFLP fingerprint profile, differences in the translation elongation factor 1-α and β-tubulin DNA sequence and interspecies mating compatibility analyses. Intraspecies mating compatibility suggests that sexual reproduction might be common for field isolates of F. temperatum, and reliable female fertile mating population tester strains were proposed for this heterothallic species.

Molecular strategy for identification in Aspergillus section Flavi

Aspergillus flavus is one of the most common contaminants that produces aflatoxins in foodstuffs. It is also a human allergen and a pathogen of animals and plants. Aspergillus flavus is included in the Aspergillus section Flavi that comprises 11 closely related species producing different profiles of secondary metabolites. A six-step strategy has been developed that allows identification of nine of the 11 species. First, three real-time PCR reactions allowed us to discriminate four groups within the section: (1) A. flavus/Aspergillus oryzae/Aspergillus minisclerotigenes/Aspergillus parvisclerotigenus; (2) Aspergillus parasiticus/Aspergillus sojae/Aspergillus arachidicola; (3) Aspergillus tamarii/Aspergillus bombycis/Aspergillus pseudotamarii; and (4) Aspergillus nomius. Secondly, random amplification of polymorphic DNA (RAPD) amplifications or SmaI digestion allowed us to differentiate (1) A. flavus, A. oryzae and A. minisclerotigenes; (2) A. parasiticus, A. sojae and A. arachidicola; (3) A. tamarii, A. bombycis and A. pseudotamarii. Among the 11 species, only A. parvisclerotigenus cannot be differentiated from A. flavus. Using the results of real-time PCR, RAPD and SmaI digestion, a decision-making tree was drawn up to identify nine of the 11 species of section Flavi. In contrast to conventional morphological methods, which are often time-consuming, the molecular strategy proposed here is based mainly on real-time PCR, which is rapid and requires minimal handling.

Gibberella musae (Fusarium musae) sp. nov., a recently discovered species from banana is sister to F. verticillioides

Several strains of Fusarium isolated from banana were identified previously as F. verticillioides (Sacc.) Nirenberg but described as unable to produce fumonisin. Here we report biochemical and morphological evidence, as well as multilocus phylogenetic analyses based on elongation factor (EF-1α), calmodulin, β-tubulin, and the second largest subunit of RNA polymerase II (RPB2) sequences, indicating that these isolates represent a unique lineage in the Gibberella fujikuroi species complex related to but distinct from F. verticillioides. Together with previous results of molecular studies, as well as with results of metabolite analyses, crossing experiments, pathogenicity tests and morphological characterization, these new data indicate that these strains isolated from banana represent a new species, Gibberella musae Van Hove et al. sp. nov. (anamorph: Fusarium musae Van Hove et al. sp. nov.), which is described herein.

Biodiversity of Fusarium species in ears and stalks of maize plants in Belgium

In order to investigate the pre-harvest contamination of maize plants by Fusarium species in Belgium, a three-year survey has been performed in five fields in which three hybrids differing in susceptibility to maize stalk rot were sampled at four different physiological stages. An extensive collection of 5,659 Fusarium isolates characterized at the species level was established during the 2005, 2006, and 2007 growing seasons, with a total of 23 different Fusarium species identified to occur on ears and stalks. A high number of plants was already contaminated by Fusarium spp. at the anthesis stage, although no symptoms were visible on ears or on stalks. As the season progressed, the incidence of Fusarium-infected maize plants reached 100% in several fields. At the end of the growing season, the most frequently isolated species in maize ears were F. graminearum, sometimes associated with F. avenaceum, F. crookwellense, F. culmorum, F. poae, and F. temperatum, a new species recently described on maize. The predominant Fusarium species detected in stalks at the end of the growing season were F. graminearum and F. crookwellense, often associated with F. culmorum and F. temperatum. Year-to-year variability observed for the incidence of F. graminearum can most likely be associated with differences in climatic conditions among the three years.

The compositional mosaic of Fusarium species and their mycotoxins in unprocessed cereals, food and feed products in Belgium

Global food safety depends on continuous monitoring of food contaminants such as mycotoxins in cereals and cereal-derived products. Here, we combine this type of investigation with quantitative occurrence data on Fusarium infestation of these products in extensive correlation studies. Finally, this contributes to a thorough understanding of the presence, origin and physiology of Fusarium Head Blight (FHB) related mycotoxins and the correlations within their ranks. Two hundred and thirty-seven samples were analyzed from diverse cereal matrices, representing the most important stages of the cereal food and feed chain in Belgium. Food, feed and non-processed field samples were investigated, with a strong emphasis on whole-grain food products. Two approaches were pursued to estimate the full scope of FHB and its repercussions: UPLC-MS/MS was applied to detect twelve different mycotoxins, and Q-PCR was used to measure the presence of ten Fusarium species. We found that different matrices have different characteristic contamination profiles, and extensive correlation studies identified certain mycotoxins for future assessment (e.g. moniliformin produced by the Fusarium avenaceum/Fusarium tricinctum species group). The investigated harvest year of 2012 yielded many non-processed field materials containing elevated levels of deoxynivalenol (DON), while even in a so-called DON-year less prevalent toxins such as T-2 and HT-2 might be considered problematic due to their consistent co-occurrence with related mycotoxins. Our data illustrate complex interactions between the many Fusarium species that are responsible for FHB and their mycotoxins. Correlation studies demonstrate that consistent co-occurrence of mycotoxins is not to be neglected, and pinpoint issues for future surveillance and legislation.

Genomic and pathogenic diversity in Colletotrichum gloeosporioides from wild native Mexican Stylosanthes spp., and taxonomic implications

RAPD amplifications of 119 Mexican Colletotrichum gloeosporioides isolates from wild native Stylosanthes plants, of two Australian (internal reference isolates of type A and type B) and of two African (previously demonstrated as different from the A and B types) allowed the generation of four polymorphic clusters. The RAPD cluster I contained the Australian type A and 103 Mexican isolates from other species than S. guianensis. The cluster II contained the Australian type B isolate and 14 Mexican isolates from S. guianensis. The clusters III and IV contained one African and one Mexican isolate each. A phylogenetic study was performed using the ITS1 sequences from 32 representative Mexican isolates selected in the four RAPD clusters, from the two Australian and the two African used as references, from 13 C. gloeosporioides from various hosts and origins available in the EMBL databank and from three phylogenetically related species of Colletotrichum. Two clusters (S and VHO) were generated. Cluster S contained the 30 isolates clustered in RAPD I and II, and the type A and B isolates from Stylosanthes selected in EMBL. Cluster VHO contained the two isolates belonging to RAPD clusters III and IV as well as the remaining Colletotrichum spp. and C. gloeosporioides isolates from various hosts and origins selected in EMBL. Severe type A lesions were observed on several Stylosanthes species, including S. guianensis, after inoculation with the 26 Mexican isolates selected in RAPD cluster I. The four Mexican isolates selected in RAPD cluster II induced extensive type B lesions only on S. guianensis genotypes. The virulence spectra of the isolates was not linked to variations in their ITS1 region sequence. A wide range of anthracnose lesions was induced by the Mexican isolates in interactions with the various Stylosanthes genotypes. On the basis of results obtained from the molecular and pathogenic experiments in the present study, of morphological and molecular data from previous work, as well as from other authors, a taxonomic infraspecific differentiation was proposed, as C. gloeosporioides f. stylosanthis f. sp. stylosanthis and f. sp. guianensis.

Gibberella xylarioides sensu lato from Coffea canephora: a new mating population in the Gibberella fujikuroi species complex.

ABSTRACT Gibberella xylarioides Heim & Saccas (presumed anamorph, Fusarium xylarioides Steyaert) is the causal agent of coffee wilt disease, an economically important tracheomycosis in Africa. In vitro crosses carried out with Congolese, Ugandan, and Tanzanian single-ascospore/conidial isolates originating from diseased Coffea canephora/excelsa demonstrated a heterothallic mating system, controlled by a single locus with two alleles, MAT-1 and MAT-2. Compatible isolates produced fertile perithecia within 2 to 8 weeks after mating. Mating type (MAT) was characterized by PCR with primer pairs previously developed for the Gibberella fujikuroi species complex (GFC) and for Fusarium oxysporum. All strains analyzed were morphologically identical and corresponded to Booths description of the “female” F. xylarioides strain. Based on crossing results and MAT-2/translation elongation 1-α (tef) sequence data, G. xylarioides, as currently understood, is demonstrated to encompass at least three “groups”: G. xylarioides sensu strictu Ia, defined hitherto by two “historical” West African strains originating from the severe 1930s to 1950s epidemic (CBS 25852 and CBS 74979); G. xylarioides sensu strictu Ib, defined by two “historical” Central African lowland strains (DSMZ 62457 and ATCC 15664); and G. xylarioides sensu lato II, containing Congolese, Ugandan, and Tanzanian C. canephora/excelsa isolates. Infertility of crosses between the coffee wilt pathogen and known GFC mating populations demonstrates that G. xylarioides sensu lato constitutes a new biological species within the G. fujikuroi complex. MUCL 44532/MUCL 43887 and MUCL 35223/MUCL 44549 are proposed as G. xylarioides sensu lato II MAT-1/MAT-2 reference mating type tester strains.