ABSTRACT
A novel negative-stranded RNA mycovirus was isolated from the phytopathogenic fungus Fusarium sibiricum strain AH32. This virus, tentatively named "Fusarium sibiricum coguvirus 1" (FsCV1), has a bipartite genome consisting of two RNA segments (RNA1 and RNA2). The negative-sense RNA1 is 6711 nt in length, encoding the RNA-dependent RNA polymerase (RdRp, p251) in the viral complementary (vc) strand. The ambisense RNA2 (1204 nt long) encodes two proteins from overlapping genes: the nucleocapsid protein (NP, p38) in the vc strand and a protein of unknown function (UFP, p36) in the viral (v) strand. In contrast to other Bunyavirales members, in FsCV1, the two open reading frames are separated by a long AU-rich intergenic region (IR). Sequence comparisons and phylogenetic analysis based on RdRp and NP sequences demonstrated that FsCV1 is a novel bipartite negative-stranded RNA mycovirus of the genus Coguvirus, family Phenuiviridae, order Bunyavirales.
Subject(s)
Fungal Viruses , Fusarium , RNA Viruses , Phylogeny , RNA, Viral/genetics , RNA-Dependent RNA Polymerase/genetics , RNA, Double-Stranded/genetics , Open Reading Frames , Genome, ViralABSTRACT
Scientific communication is facilitated by a data-driven, scientifically sound taxonomy that considers the end-user's needs and established successful practice. In 2013, the Fusarium community voiced near unanimous support for a concept of Fusarium that represented a clade comprising all agriculturally and clinically important Fusarium species, including the F. solani species complex (FSSC). Subsequently, this concept was challenged in 2015 by one research group who proposed dividing the genus Fusarium into seven genera, including the FSSC described as members of the genus Neocosmospora, with subsequent justification in 2018 based on claims that the 2013 concept of Fusarium is polyphyletic. Here, we test this claim and provide a phylogeny based on exonic nucleotide sequences of 19 orthologous protein-coding genes that strongly support the monophyly of Fusarium including the FSSC. We reassert the practical and scientific argument in support of a genus Fusarium that includes the FSSC and several other basal lineages, consistent with the longstanding use of this name among plant pathologists, medical mycologists, quarantine officials, regulatory agencies, students, and researchers with a stake in its taxonomy. In recognition of this monophyly, 40 species described as genus Neocosmospora were recombined in genus Fusarium, and nine others were renamed Fusarium. Here the global Fusarium community voices strong support for the inclusion of the FSSC in Fusarium, as it remains the best scientific, nomenclatural, and practical taxonomic option available.
Subject(s)
Fusarium , Fusarium/genetics , Phylogeny , Plant Diseases , PlantsABSTRACT
The biological characterization of Microdochium majus, M. nivale, and M. seminicola strains with wide geographical origins showed the diversity of their pathogenic properties and metabolite compounds, allowing them to exist in their habitats. Significant differences in the ability of Microdochium fungi to cause lesions on wheat and oat leaves were found. The intensity of symptoms depended on the species and substrate origin of the strains. On average M. seminicola strains were able to cause less leaf necrosis than M. majus and M. nivale. The volatile organic compound (VOC) profile of Microdochium fungi included 29 putative fungal metabolites. The spectrum of the identified VOCs in M. seminicola strains was much richer than that in M. majus and M. nivale strains. In addition, the strains of M. seminicola emitted at least six sesquiterpenes. Mycotoxin analysis by HPLC/MS/MS revealed that the analyzed Microdochium strains did not produce any toxic metabolites typically produced by filamentous fungi.
Subject(s)
Ascomycota/physiology , Ascomycota/metabolism , Ascomycota/pathogenicity , Chromatography, High Pressure Liquid/methods , Mycotoxins/metabolism , Plant Diseases/microbiology , Tandem Mass Spectrometry/methods , Volatile Organic Compounds/metabolismABSTRACT
The compositions of volatile organic compounds (VOCs) emitted by Fusarium fungi (F. langsethiae, F. sibiricum, F. poae, and F. sporotrichioides) grown on two nutritive substrates: potato sucrose agar (PSA) and autoclaved wheat kernels (WK) were investigated. The culturing of fungi and study of their VOC emissions were performed in chromatographic vials at room temperature (23 - 24 °C) and the VOCs were sampled by a solid-phase microextraction on a 85 µm carboxen/polydimethylsiloxane fiber. GC/MS was performed using a 60-m HP-5 capillary column. Components of the VOC mixture were identified by electron impact mass spectra and chromatographic retention indices (RIs). The most abundant components of the VOC mixture emitted by Fusarium fungi are EtOH, AcOH, (i) BuOH, 3-methylbutan-1-ol, 2-methylbutan-1-ol, ethyl 3-methylbutanoate, terpenes with M 136, sesquiterpenes with M 204 (a total of about 25), and trichodiene. It was found that the strains grown on PSA emit a wider spectrum and larger amount of VOCs compared with those grown on wheat kernels. F. langsethiae strain is the most active VOC producer on both substrates. The use of SPME and GC/MS also offers the potential for differentiation of fungal species and strains.
Subject(s)
Fusarium/growth & development , Fusarium/metabolism , Gases/analysis , Volatile Organic Compounds/analysis , Agar/metabolism , Gas Chromatography-Mass Spectrometry , Gases/chemistry , Gases/metabolism , Seeds/metabolism , Solanum tuberosum/chemistry , Solid Phase Microextraction , Sucrose/metabolism , Triticum , Volatile Organic Compounds/chemistry , Volatile Organic Compounds/metabolism , VolatilizationABSTRACT
Enzymes capable of modifying the sulfated polymeric molecule of fucoidan are mainly produced by different groups of marine organisms: invertebrates, bacteria, and also some fungi. We have discovered and identified a new strain of filamentous fungus Fusarium proliferatum LE1 (deposition number in Russian Collection of Agricultural Microorganisms is RCAM02409), which is a potential producer of fucoidan-degrading enzymes. The strain LE1 (RCAM02409) was identified on the basis of morphological characteristics and analysis of ITS sequences of ribosomal DNA. During submerged cultivation of F. proliferatum LE1 in the nutrient medium containing natural fucoidan sources (the mixture of brown algae Laminaria digitata and Fucus vesiculosus), enzymic activities of α-L-fucosidase and arylsulfatase were inducible. These enzymes hydrolyzed model substrates, para-nitrophenyl α-L-fucopyranoside and para-nitrophenyl sulfate, respectively. However, the α-L-fucosidase is appeared to be a secreted enzyme while the arylsulfatase was an intracellular one. No detectable fucoidanase activity was found during F. proliferatum LE1 growth in submerged culture or in a static one. Comparative screening for fucoidanase/arylsulfatase/α-L-fucosidase activities among several related Fusarium strains showed a uniqueness of F. proliferatum LE1 to produce arylsulfatase and α-L-fucosidase enzymes. Apart them, the strain was shown to produce other glycoside hydrolyses.
Subject(s)
Arylsulfatases/metabolism , Fusarium/metabolism , Polysaccharides/metabolism , alpha-L-Fucosidase/metabolism , DNA, Ribosomal , Fucus , Fusarium/growth & development , Fusarium/isolation & purification , Laminaria , Nitrobenzenes/metabolism , Nitrophenols/metabolism , Sequence AnalysisABSTRACT
Dry rot of potato tubers is a harmful disease caused by species of the Fusarium genus. Studies on the composition and features of Fusarium spp. that cause the disease in Russia are limited. Thirty-one Fusarium strains belonging to the F. sambucinum species complex (FSAMSC) and F. solani species complex (FSSC) were accurately identified using multilocus phylogenetic analysis of the tef and rpb2 loci, and their physiological characteristics were studied in detail. As a result, 21 strains of F. sambucinum s. str. and 1 strain of F. venenatum within the FSAMSC were identified. Among the analyzed strains within the FSSC, one strain of F. mori, four strains of F. noneumartii, and two strains of both F. stercicola and F. vanettenii species were identified. This is the first record of F. mori on potato as a novel host plant, and the first detection of F. noneumartii and F. stercicola species in Russia. The clear optimal temperature for the growth of the strains belonging to FSAMSC was noted to be 25 °C, with a growth rate of 11.6-15.0 mm/day, whereas, for the strains belonging to FSSC, the optimal temperature range was between 25 and 30 °C, with a growth rate of 5.5-14.1 mm/day. The distinctive ability of F. sambucinum strains to grow at 5 °C has been demonstrated. All analyzed Fusarium strains were pathogenic to potato cv. Gala and caused extensive damage of the tuber tissue at an incubation temperature of 23 °C for one month. Among the fungi belonging to the FSAMSC, the F. sambucinum strains were more aggressive and caused 23.9 ± 2.2 mm of necrosis in the tubers on average compared to the F. venenatum strain-17.7 ± 1.2 mm. Among the fungi belonging to the FSSC, the F. noneumartii strains were the most aggressive and caused 32.2 ± 0.8 mm of necrosis on average. The aggressiveness of the F. mori, F. stercicola, and especially the F. vanettenii strains was significantly lower: the average sizes of damage were 17.5 ± 0.5 mm, 17.2 ± 0.2 mm, and 12.5 ± 1.7 mm, respectively. At an incubation temperature of 5 °C, only the F. sambucinum strains caused tuber necroses in the range of 6.7 ± 0.5-15.9 ± 0.8 mm.
ABSTRACT
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.
Subject(s)
Fusarium/classification , Plants/microbiology , Fusarium/genetics , Phylogeny , Plant Diseases/microbiologyABSTRACT
A Fusarium species associated with sunflower based on multilocus genealogy, morphological, physiological, ecological, mating type, and mycotoxin production data is formally described as the newly discovered species Fusarium bilaiae. The F. bilaiae strains formed a genealogically exclusive lineage within the African clade of the F. fujikuroi species complex. Comparison of morphological characteristics of F. bilaiae strains with those of the closely related F. phyllophilum strain NRRL 13617 revealed similarities in the main micromorphology of both species: production of numerous one-celled microconidia in false heads and short chains on monophialides and polyphialides and the absence of macroconidia and sporodochia. There was a slight but significant distinction between the two species when the strains were grown on different agar media, as well as in the shape and width of microconidia. Fusarium bilaiae strains isolated from symptomatic sunflower were not pathogenic to members of the Asteraceae tested; apparently, they live as saprophytes or endophytes in sunflower tissues. A difference between the strains of the two species in the production of mycotoxins was demonstrated with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis. On autoclaved rice, F. bilaiae did not produce fumonisins and beauvericin but produced moniliformin, whereas F. phyllophilum produced all these mycotoxins. A polymerase chain reaction (PCR) assay specific for mating type alleles identified F. bilaiae as a putative heterothallic species with MAT1-1 and MAT1-2 idiomorphs, but laboratory crosses were unsuccessful. Determining the area and host range of the new endophytic species F. bilaiae is a priority for future research.
Subject(s)
Asteraceae , Fusarium , Helianthus , Mycotoxins , Fusarium/genetics , Tandem Mass SpectrometryABSTRACT
Fusarium langsethiae is a suspected plant-pathogenic fungus causing cereal contamination with trichothecene mycotoxins. Here, we report the genome sequences of two F. langsethiae strains, MFG217701 (a prototroph) and MFG217702 (a biotin auxotroph), isolated from a grain of oat harvested in Russia.
ABSTRACT
The first appearance of Fusarium head blight (FHB)-and the beginning of scientific research of this disease-occurred the Far East region of Russia at the end of the 19th century. In the summer of 2019, in the Amur region, which comprises 60-70% of grain production in the Russian Far East, flooding caused a state of emergency. The quality of wheat and barley grains grown under natural conditions of FHB outbreaks, including grain infection, fungal species composition, DNA content of F. graminearum and chemotypes, and the presence of various mycotoxins, was studied. Fusarium infection rates reached extremely high percentages, 51-98%, the majority of which were F. graminearum infections. The amount of F. graminearum DNA in wheat grain samples was higher than in the barley grain samples and averaged 6.1 and 2.1 pg/ng, respectively. The content of deoxynivalenol (DON) in the wheat samples reached 13,343 ppb and in barley reached 7,755 ppb. A multilocus genotyping assay was conducted on the partially sequenced fragments of the translation elongation factor EF-1a, ammonium ligase gene, reductase gene, and 3-O-acetyltransferase gene in 29 Fusarium graminearum sensu lato strains from the grain harvested in the Amur region. All strains from the Far East region were characterized as F. graminearum sensu stricto; 70% were the 15-AcDON chemotype, while the other strains were the 3-AcDON chemotype. According to the results, after 140 years of study of FHB, we are still not very successful in controlling this disease if conditions are favorable for pathogen development. Even at present, some of the grain harvested must be destroyed, as high contamination of mycotoxins renders it unusable.
ABSTRACT
Alternaria fungi dominate the grain microbiota in many regions of the world; therefore, the detection of species that are able to produce mycotoxins has received much attention. A total of 178 grain samples of wheat, barley and oat obtained from the Urals and West Siberia regions of Russia in 2017-2019 were included in the study. Grain contamination with Alternaria fungi belonging to sections Alternaria and Infectoriae was analysed using qPCR with specific primers. The occurrence of four mycotoxins produced by Alternaria, AOH, AME, TEN, and TeA, was defined by HPLC-MS/MS. Alternaria DNA was found in all analysed grain samples. The prevalence of DNA of Alternaria sect. Alternaria fungi (range 53 × 10-4-21,731 × 10-4 pg/ng) over the DNA of Alternaria sect. Infectoriae (range 11 × 10-4â4237 × 10-4 pg/ng) in the grain samples was revealed. Sixty-two percent of grain samples were contaminated by at least two Alternaria mycotoxins. The combination of TEN and TeA was found most often. Eight percent of grain samples were contaminated by all four mycotoxins, and only 3% of samples were free from the analysed secondary toxic metabolites. The amounts varied in a range of 2-53 µg/kg for AOH, 3-56 µg/kg for AME, 3-131 µg/kg for TEN and 9-15,000 µg/kg for TeA. To our knowledge, a new global maximum level of natural contamination of wheat grain with TeA was detected. A positive correlation between the amount of DNA from Alternaria sect. Alternaria and TeA was observed. The significant effects of cereal species and geographic origin of samples on the amounts of DNA and mycotoxins of Alternaria spp. in grain were revealed. Barley was the most heavily contaminated with fungi belonging to both sections. The content of AOH in oat grain was, on average, higher than that found in wheat and barley. The content of TEN in the grain of barley was lower than that in wheat and similar to that in oat. The content of TeA did not depend on the cereal crop. The effect of weather conditions (summer temperature and rainfall) on the final fungal and mycotoxin contamination of grain was discussed. The frequent co-occurrence of different Alternaria fungi and their mycotoxins in grain indicates the need for further studies investigating this issue.
Subject(s)
Alternaria/isolation & purification , Edible Grain/microbiology , Mycotoxins/analysis , Alternaria/genetics , Avena/microbiology , DNA, Fungal/isolation & purification , Food Contamination/analysis , Hordeum/microbiology , Russia , Triticum/microbiology , WeatherABSTRACT
In total, 46 Microdochium strains from five different geographic regions of Russia were explored with respect to genetic diversity, morphology, and secondary metabolites. Based on the results of PCR, 59% and 28% of the strains were identified as M. nivale and M. majus, respectively. As a result of sequencing four genome regions, namely ITS, LSU, BTUB, and RPB2 (2778 bp), five genetically and phenotypically similar strains from Western Siberia were identified as M. seminicola, which, according to our findings, is the prevalent Microdochium species in this territory. This is the first record of M. seminicola in Russia. Attempts were made to distinguish between Microdochium species and to identify species-specific morphological characteristics in the anamorph and teleomorph stages and physiological properties. We examined the occurrence frequency of conidia with different numbers of septa in the strains of Microdochium. The predominance of three-septate macroconidia in M. majus was higher than that in M. nivale and typically exceeded 60% occurrence. Most M. majus and M. nivale strains formed walled protoperithecia on wheat stems. Only three strains of M. majus and one strain each of M. nivale and M. seminicola produced mature perithecia. The growth rate of M. seminicola strains was significantly lower on agar media at 5-25 °C than those of M. majus and M. nivale strains. Multimycotoxin analysis by HPLC-MS/MS revealed that the strains of three Microdochium species did not produce any toxic metabolites.
ABSTRACT
We investigated Fusarium graminearum complex (Fg complex) species diversity and toxin potential in European and Asian regions of the Russian Federation and adjoining regions northwest to Finland and south near Harbin, Heilongjiang Province, China, to expand our knowledge of the host range and geographic distribution of these economically devastating cereal head blight pathogens. Results of a recently described multilocus genotyping (MLGT) assay revealed that F. graminearum was the sole Fg complex pathogen in northern Europe and the predominant one in Asia (90.5%). Even though isolates of F. graminearum were segregating for 3-acetyldeoxynivalenol (3ADON) and 15-acetyldeoxynivalenol (15ADON) chemotype in nearly equal frequencies in the regions sampled on both continents, significant differences in the geographic distribution of isolates producing these acetyl ester derivatives of deoxynivalenol (DON) were observed in Europe. While 93.5% of the isolates in southern Russia (n = 43 of 46) possessed the 15ADON chemotype, isolates of F. graminearum recovered in Finland and northwestern Russia (n = 40) were exclusively 3ADON producers. Based on results of the MLGT assay, species identity of 10 genetically novel Fg complex isolates from the Russian Far East was investigated further via molecular phylogenetic analyses of multilocus DNA sequence data. Results of these analyses resolved these isolates as a phylogenetically distinct, reciprocally monophyletic sister lineage of F. asiaticum, which together with F. vorosii form a newly discovered Asian clade within the Fg complex. Because this novel lineage fulfills the highly conservative criterion of genealogical exclusivity under phylogenetic species recognition it is formally described herein as F. ussurianum. In addition to morphologically characterizing isolates of F. ussurianum, experiments were conducted to assess pathogenicity to wheat and trichothecene toxin potential in planta.
Subject(s)
Edible Grain/microbiology , Fusarium/classification , Plant Diseases/microbiology , Asia , DNA, Fungal/analysis , DNA, Fungal/genetics , Europe , Fusarium/genetics , Fusarium/metabolism , Genetic Variation , Geography , Phylogeny , Sequence Analysis, DNA , Species Specificity , Trichothecenes/biosynthesisABSTRACT
Wheat grains collected in three regions of Russia-Volga, Ural, and West Siberia-were analyzed for triangulation of methods in analysis of toxigenic Fusarium species. The presence of fungi and quantitative content of their biomass were detected by using various analytical methods, including a mycological and immunochemical methods, and quantitative PCR. Additionally, an enzyme-linked immunosorbent assay and high-performance liquid chromatography with tandem mass spectrometry were applied for determination of mycotoxins. Regional differences were found regarding the contamination of wheat grain by Fusarium fungi and their toxins. The most important observation was the detection of F. graminearum in the Ural and West Siberian regions, where this pathogen had not been found previously. A maximum damaged grains by F. graminearum and F. sporotrichioides was found in the grain samples from West Siberia. The DNA of F. graminearum was detected in 19.2% and DNA of F. sporotrichioides was found in 84.1% of the analyzed grain samples. The amount of Fusarium antigens in the grain samples from the West Siberian region was 7-8 times higher than in the grain samples from the other two regions. Significant contamination of the grain with deoxynivalenol and T-2/HT-2 toxins (maximum contents were 2239 ppb and 199 ppb, respectively) was detected in the West Siberian region.
Subject(s)
Edible Grain/microbiology , Fusarium , Triticum/microbiology , Antigens, Fungal/analysis , DNA, Fungal/analysis , Edible Grain/chemistry , Environmental Monitoring , Food Contamination/analysis , Fusarium/genetics , Mycotoxins/analysis , Russia , Triticum/chemistryABSTRACT
Trichothecenes are sesquiterpenoid mycotoxins produced by fungi from the order Hypocreales, including members of the Fusarium genus that infect cereal grain crops. Different trichothecene-producing Fusarium species and strains have different trichothecene chemotypes belonging to the Type A and B class. These fungi cause a disease of small grain cereals, called Fusarium head blight, and their toxins contaminate host tissues. As potent inhibitors of eukaryotic protein synthesis, trichothecenes pose a health risk to human and animal consumers of infected cereal grains. In 2009, Foroud and Eudes published a review of trichothecenes in cereal grains for human consumption. As an update to this review, the work herein provides a comprehensive and multi-disciplinary review of the Fusarium trichothecenes covering topics in chemistry and biochemistry, pathogen biology, trichothecene toxicity, molecular mechanisms of resistance or detoxification, genetics of resistance and breeding strategies to reduce their contamination of wheat and barley.
Subject(s)
Edible Grain , Food Contamination/analysis , Trichothecenes , Animal Feed/analysis , Animal Feed/microbiology , Edible Grain/microbiology , Fungal Proteins/genetics , Fusarium/genetics , Fusarium/growth & development , Fusarium/metabolism , Hordeum/microbiology , Plant Diseases/microbiology , Trichothecenes/analysis , Trichothecenes/metabolism , Triticum/microbiologyABSTRACT
Fusarium avenaceum and closely related species are common fungi on various plants, cultivated in different climatic regions. The aim of this study was to determine the taxonomic affiliations of the F. avenaceum, Fusarium arthrosporioides, and Fusarium anguioides strains by using morphological, physiological and molecular-genetic approaches. Twenty-six single-spored morphologically identified strains, which were mainly from cereals, were investigated in order to find out, if they belong to a separate species. Pathogenicity of strains to wheat seedlings and ISSR (Inter Simple Sequence Repeats) fingerprint and beta-tubulin DNA sequence patterns were analyzed. According to phylogenetic analyses, the strains could be divided into two big groups consisting of mostly F. avenaceum or F. anguioides strains. F. arthrosporioides was not detected as a separate species by the sum of the characters. F. anguioides was characterized as a separate species, which could be identified by morphological and molecular data. High genetic diversity of the F. avenaceum and related species was revealed. One F. anguioides strain (rudbeckia, Vladivostok, Russia), had an identical beta-tubulin sequence with two previously sequenced strains of Fusarium tricinctum species complex, which were isolated from dicotyledonous plants in Asia.
ABSTRACT
Analysis of 49 strains of Fusarium langsethiae originating from northern Europe (Russia, Finland, Sweden, UK, Norway, and Latvia) revealed the presence of spontaneous auxotrophic mutants that reflect natural intraspecific diversity. Our investigations detected that 49.0% of F. langsethiae strains were auxotrophic mutants for biotin, and 8.2% of the strains required thiamine as a growth factor. They failed to grow on vitamin-free media. For both prototrophic and auxotrophic strains, no growth defect was observed in rich organic media. Without essential vitamins, a significant reduction in the growth of the auxotrophic strains results in a decrease of the formation of T-2 toxin and diacetoxyscirpenol. In addition, all analysed F. langsethiae strains were distinguished into two subgroups based on PCR product sizes. According to our results, 26 and 23 strains of F. langsethiae belong to subgroups I and II respectively. We determined that the deletion in the intergenic spacer (IGS) region of the rDNA of F. langsethiae belonging to subgroup II is linked with temperature sensitivity and causes a decrease in strain growth at 30 °C. Four thiamine auxotrophic strains were found in subgroup I, while 21 biotin auxotrophic strains were detected in subgroups II. To the best of our knowledge, the spontaneous mutations in F. langsethiae observed in the present work have not been previously reported.
ABSTRACT
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.
ABSTRACT
Production of type A trichothecenes has been reported in the closely related species Fusarium langsethiae and F. sporotrichioides. Here, we characterized a collection of Fusarium isolates from Siberia and the Russian Far East (hereafter Asian isolates) that produce high levels of the type A trichothecene T-2 toxin and are similar in morphology to the type A trichothecene-producing F. langsethiae, and to F. poae which often produces the type B trichothecene nivalenol. The Asian isolates possess unique macroscopic and microscopic characters and have a unique TG repeat in the nuclear ribosomal intergenic spacer (IGS rDNA) region. In Asian isolates, the TRI1-TRI16 locus, which determines type A versus type B trichothecene production in other species, is more similar in organization and sequence to the TRI1-TRI16 locus in F. sporotrichioides and F. langsethiae than to that in F. poae. Phylogenetic analysis of the TRI1 and TRI16 gene coding regions indicates that the genes in the Asian isolates are more closely related to those of F. sporotrichioides than F. langsethiae. Phylogenetic analysis of the beta-tubulin, translation elongation factor, RNA polymerase II and phosphate permease gene sequences resolved the Asian isolates into a well-supported sister lineage to F. sporotrichioides, with F. langsethiae forming a sister lineage to F. sporotrichioides and the Asian isolates. The Asian isolates are conspecific with Norwegian isolate IBT 9959 based on morphological and molecular analyses. In addition, the European F. langsethiae isolates from Finland and Russia were resolved into two distinct subgroups based on analyses of translation elongation factor and IGS rDNA sequences. Nucleotide polymorphisms within the IGS rDNA were used to design PCR primers that successfully differentiated the Asian isolates from F. sporotrichioides and F. langsethiae. Based on these data, we formally propose that the Asian isolates together with Norwegian isolate IBT 9959 comprise a novel phylogenetic species, F. sibiricum, while the two subgroups of F. langsethiae only represent intraspecific groups.