Fusarium graminearum Species Complex: A Bibliographic Analysis and Web-Accessible Database for Global Mapping of Species and Trichothecene Toxin Chemotypes.

Fusarium graminearum is ranked among the five most destructive fungal pathogens that affect agroecosystems. It causes floral diseases in small grain cereals including wheat, barley, and oats, as well as maize and rice. We conducted a systematic review of peer-reviewed studies reporting species within the F. graminearum species complex (FGSC) and created two main data tables. The first contained summarized data from the articles including bibliographic, geographic, methodological (ID methods), host of origin and species, while the second data table contains information about the described strains such as publication, isolate code(s), host/substrate, year of isolation, geographical coordinates, species and trichothecene genotype. Analyses of the bibliographic data obtained from 123 publications from 2000 to 2021 by 498 unique authors and published in 40 journals are summarized. We describe the frequency of species and chemotypes for 16,274 strains for which geographical information was available, either provided as raw data or extracted from the publications, and sampled across six continents and 32 countries. The database and interactive interface are publicly available, allowing for searches, summarization, and mapping of strains according to several criteria including article, country, host, species and trichothecene genotype. The database will be updated as new articles are published and should be useful for guiding future surveys and exploring factors associated with species distribution such as climate and land use. Authors are encouraged to submit data at the strain level to the database, which is accessible at https://fgsc.netlify.app.

First report of Pyrenophora teres f. teres causing leaf spot on wheat in Brazil.

Pyrenophora teres f. teres is well known for causing net form net blotch on barley, but its pathogenicity on wheat has also been reported in Hungary (Tóth et al. 2008) and east Russia (Mikhailova et al. 2010). The report from Hungary mentioned symptoms of dark brown necrotic lesions with or without chlorotic margins, while the report from Russia described symptoms similar to tan spot. A survey of fungal pathogens causing leaf spot diseases on wheat in the humid subtropical areas of Paraná State, southern Brazil, was conducted in 2016. Twenty four isolates, representing different wheat fields in the municipalities of Guarapuava and Pinhão, in the state of Paraná, were collected from wheat plants at the soft dough stage. The isolates analyzed in our study were obtained from leaf fragments of wheat cv. Toruk and were removed from the edge of lesions resembling tan spot, a major fungal disease of wheat in that region, caused by Pyrenophora tritici-repentis. The leaves presented oval, diamond, or elongate-shape lesions with tan necrotic spots. Still, they did not always show the brown center, which seemed not very characteristics of tan spot caused by P. tritici-repentis. Yellow halos surrounded the necrotic lesions. The fragments were superficially disinfected for 30 sec in 0.5% sodium hypochlorite, washed in sterile water, dried with a sterile paper towel, and transferred to petri plates containing potato dextrose agar (PDA) medium. After four days of incubation at 24±2ºC and a photoperiod of 12 h, hyphal tips were removed from the PDA medium and transferred to petri plates containing V-8 medium. Plates were incubated for 14 days in alternate cycles of 12 h at 23±2ºC under light and 12 h at 19±2ºC in darkness for sporulation. Pure cultures were obtained from single conidia produced on V-8 medium. Single spore isolates, representing different fields, were obtained from individual leaves. Conidia measuring 38-82 µm × 16-20 µm were smoothly cylindrical and straight, with rounded ends, subhyaline or yellowish brown, with four to seven pseudosepta. Genomic DNA extracted from cultures grown in yeast extract broth medium was used for PCR amplification of the ribosomal internal transcribed spacers and the 5.8S ribosomal DNA gene (ITS-5.8S rDNA) using the primers ITS4 and ITS5 described by White et al. (1990). A BLAST search of the GenBank database revealed that the ITS-5.8S rDNA sequence fragments (505 bp) from 12 out of 67 isolates were 99-100% similar to P. teres accession number EF452452 (Andrie et al. 2008). The DNA sequences of three P. teres isolates (20C3, 20A3, and 20A1) were deposited in GenBank (accession nos. MH594043.1, MH594042.1, and MH594041.1, respectively). These isolates were obtained from different fields in the municipality of Pinhão (25° 42 '26 "S; 51° 38' 0" W), in Paraná State. To test the pathogenicity of the isolates 20C3, 20A3, and 20A1, a conidial suspension for each isolate with 2-3 × 103 conidia per mL was sprayed at the boot stage (BBCH-scale 45) on leaves of wheat cv. Toruk and on susceptible barley cv. Ana 01 plants growing in pots. Each pot containing 5-7 plants was considered as replicate. Wheat plants were also inoculated with a P. tritici-repentis isolate with a conidial suspension at the same concentration described above to serve as reference of symptoms for tan spot. Wheat and barley plants were also inoculated with sterile water to serve as a control. Six replications were inoculated for each treatment. After inoculation, the plants were maintained in a dew chamber for 24 h and then moved to a growth room at 20±2ºC and 16 h photoperiod. Initially, the leaves of wheat plants inoculated with P. teres exhibited small, irregular, dark lesions surrounded by yellow halos, and later exhibited oval to elongate, pale brown, necrotic lesions, and yellow halos. Differently from tan spot caused by P. tritici-repentis, the wheat leaves inoculated with P. teres f. teres presented a more diffuse yellowing, and the center of necrotic lesions was pale brown. The symptoms observed in the barley leaves were the net form net blotch, which allowed us to infer that the disease was caused by P. teres f. teres isolates. All inoculated plants presented symptoms, with some variation among them. Pyrenophora teres f. teres was reisolated from the inoculated wheat and barley plants. This is the first report of P. teres f. teres causing leaf disease on wheat in Brazil. The finding contributes to a better understanding of the etiology of wheat leaf spots in humid subtropical areas of southern Brazil.

Regional and field-specific factors affect the composition of fusarium head blight pathogens in subtropical no-till wheat agroecosystem of Brazil.

A multiyear survey of >200 wheat fields in Paraná (PR) and Rio Grande do Sul (RS) states was conducted to assess the extent and distribution of Fusarium graminearum species complex (FGSC) diversity in the southern Brazilian wheat agroecosystem. Five species and three trichothecene genotypes were found among 671 FGSC isolates from Fusarium head blight (FHB)-infected wheat heads: F. graminearum (83%) of the 15-acetyldeoxynivalenol (15-ADON) genotype, F. meridionale (12.8%) and F. asiaticum (0.4%) of the nivalenol (NIV) genotype, and F. cortaderiae (2.5%) and F. austroamericanum (0.9%) with either the NIV or the 3-ADON genotype. Regional differences in FGSC composition were observed, with F. meridionale and the NIV type being significantly (P<0.001) more prevalent in PR (>28%) than in RS (≤9%). Within RS, F. graminearum was overrepresented in fields below 600 m in elevation and in fields with higher levels of FHB incidence (P<0.05). Species composition was not significantly influenced by previous crop or the stage of grain development at sampling. Habitat-specific differences in FGSC composition were evaluated in three fields by characterizing a total of 189 isolates collected from corn stubble, air above the wheat canopy, and symptomatic wheat kernels. Significant differences in FGSC composition were observed among these habitats (P<0.001). Most strikingly, F. meridionale and F. cortaderiae of the NIV genotype accounted for the vast majority (>96%) of isolates from corn stubble, whereas F. graminearum with the 15-ADON genotype was dominant (>84%) among isolates from diseased wheat kernels. Potential differences in pathogenic fitness on wheat were also suggested by a greenhouse competitiveness assay in which F. graminearum was recovered at much higher frequency (>90%) than F. meridionale from four wheat varieties inoculated with an equal mixture of F. graminearum and F. meridionale isolates. Taken together, the data presented here suggest that FGSC composition and, consequently, the trichothecene contamination in wheat grown in southern Brazil is influenced by host adaptation and pathogenic fitness. Evidence that F. meridionale and F. cortaderiae with the NIV genotype are regionally significant contributors to FHB may have significant implications for food safety and the economics of cereal production.

Phytophthora acaciae sp. nov., a new species causing gummosis of black wattle in Brazil.

A new Phytophthora species was found associated with gummosis in black wattle plantations in the subtropical, humid, south of Brazil. The new species Phytophthora acaciae is formally named herein based on phylogenetic and morphological analyses. This is the fourth Phytophthora species found from this pathogen complex in black wattle plantations causing gummosis in Brazil. The other three species are P. nicotianae, P. boehmeriae, and P. frigida. Phytophthora acaciae is heterothallic with amphigynous antheridia, noncaducous, papillate sporangia and is placed in the Phytophthora clade 2 based on nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) sequences. Maximum parsimony and maximum likelihood phylogenetic analyses of P. acaciae isolates based on multigene sequences, including partial DNA sequences of three nuclear protein-coding genes (ß-tubulin, translation elongation factor-1α, and ras-related protein), two mitochondrial protein-coding genes (cytochrome c oxidase subunits I and II), in addition to ITS sequence data, support the delimitation of this new species on Acacia mearnsii from the other previously described clade 2 Phytophthora species. Pathogenicity trial confirmed that the new species causes necrotic lesions on the plant stem, with either the presence or absence of gum.

Fusarium subtropicale, sp. nov., a novel nivalenol mycotoxin-producing species isolated from barley (Hordeum vulgare) in Brazil and sister to F. praegraminearum.

Surveys were conducted in commercial wheat and barley fields in the south central production regions of state of Paraná, Brazil, from 2011 to 2015. Spikes displaying visible Fusarium head blight symptoms were collected and the pathogen isolated from the tissues. The 754 Fusarium isolates recovered were identified by a high-throughput multilocus genotyping assay (MLGT) designed to identify trichothecene toxin-producing fusaria (i.e., formerly B-clade, but referred to here as F. sambucinum species complex lineage 1 [FSAMSC-1]) together with sequencing a portion of the translation elongation factor 1-α (TEF1) gene. One strain was discovered that appeared to be closely related to but phylogenetically distinct from F. praegraminearum based on the relatively low 97.7% TEF1 identity and positive genotype obtained with one of the two F. praegraminearum species-specific MLGT probes. Molecular phylogenetic analyses of a 10-gene data set resolved this novel FSAMSC-1 species and F. praegraminearum as sisters. Formally described herein as F. subtropicale, it is phenotypically distinct from the 22 other FSAMSC-1 species in that it produces mostly 1-3-septate macroconidia. Whole-genome sequence data were used to predict its potential to produce mycotoxins. Chemical analyses confirmed that F. subtropicale could produce the mycotoxins 4,15-diacetylnivalenol, butenolide, culmorin, and fusarin C in vitro, and the pathogenicity experiment revealed that F. subtropicale could infect but not spread in susceptible hard red spring wheat cultivar "Norm."

Fusarium paranaense sp. nov., a member of the Fusarium solani species complex causes root rot on soybean in Brazil.

Isolates of Fusarium obtained from soybean plants showing symptoms of root rot collected in subtropical southern and tropical central Brazil were characterized based on phylogenetic analyses, sexual crossing, morphology, and pathogenicity tests. A novel species within the Fusarium solani species complex (FSSC) causing soybean root rot is formally described herein as Fusarium paranaense. This species can be distinguished from the other soybean root rot pathogens in the FSSC, which are commonly associated with soybean sudden death syndrome (SDS) based on analyses of the combined DNA sequences of translation elongation factor 1-α and the second largest subunit of RNA polymerase II and on interspecies mating compatibility. Bayesian and maximum parsimony phylogenetic analyses showed that isolates of F. paranaense formed a distinct group in clade 3 of the FSSC in contrast to the pathogens currently known to cause SDS, which are in clade 2. Female fertile tester strains were developed that can be used for the identification of this new species in the FSSC based on sexual crosses. All isolates were heterothallic and belonged to a distinct mating population. Fusarium tucumaniae, a known SDS pathogen, was found in the subtropical southern region of the country.