Fusarium species are common pathogens on maize and reduce the product quality through contamination with mycotoxins thus jeopardizing safety of both animal feed and human food products. Monitoring of Fusarium infected maize ears and stalks was conducted in Germany to determine the range of Fusarium species present in the field and to assess the impact of tillage, crop rotation, and weather conditions on the frequency of Fusarium species. From 2016 till 2018, a total of 387 infected ears and 190 stalk segments from 58 locations in Germany were collected. For each sample location, site-specific agronomic data on tillage and previous crops as well as meteorological data on precipitation, air temperature, and relative humidity during the vegetation period were recorded. The most frequent Fusarium species detected in maize ears were Fusarium graminearum, F. verticillioides and F. temperatum, whereas, F. graminearum, F. equiseti, F. culmorum, and F. temperatum were the species prevailing on maize stalks. Differences in the local species composition were found to be primarily associated with weather variations between the years and the microclimate at the different locations. The results indicate that mean temperature and precipitation in July, during flowering, has the strongest impact on the local range of Fusarium spp. on ears, whereas the incidence of Fusarium species on stalks is mostly affected by weather conditions during September. Ploughing significantly reduced the infection with F. graminearum and F. temperatum, while crop rotation exerted only minor effects.

Trichoderma species are widespread filamentous fungi in soils, on plant roots and decaying plant residues. Due to their strong competitiveness and mycoparasitic activity against other fungi, particular strains of Trichoderma sp. are used in agriculture as biocontrol agents against plant pathogens. Commercial products based on strains of T. harzianum or T. afroharzianum have been applied to control Rhizoctonia spp., Fusarium spp., and Phytophthora spp. in various crops. In 2018, however, severe infections of Trichoderma on maize ears were recorded for the first time in a field in Southern Germany. Infected maize cobs were sampled, the fungus was isolated in pure culture and preliminarily identified microscopically as T. harzianum. After silk channel inoculation in the greenhouse, ear rot disease of high severity was observed. In addition to fungal colonization, the dry matter content in cobs was significantly reduced compared to water inoculated cobs. In 2018 and 2019, a total of 13 T. harzianum isolates from maize cobs and maize stalks were isolated and tested, for pathogenicity on maize plants in the greenhouse, compared to several reference isolates. Four isolates proved to be highly aggressive, two biocontrol isolates, Trichodex (T39), and strain T12, induced slight infection and eleven isolates were non-pathogenic. After sequencing of the translation elongation factor-1α (tef-1α) and internal transcribes spacers (ITS), the four highly aggressive isolates were reassigned to T. afroharzianum, while the commercial biocontrol isolates Trichodex (T39), and T12, as well as the other non-pathogenic strains belonged to T. harzianum, T. atroviride, or T. tomentosum. This, to our knowledge, is the first report on Trichoderma sp. as a pathogen causing ear rot disease in maize in Europe with the potential to incite significant yield losses. We therefore propose to name this disease as "Trichoderma ear rot on maize".

Maize plants are often infected with fungal pathogens of the genus Fusarium. Taxonomic characterization of these species by microscopic examination of pure cultures or assignment to mating populations is time-consuming and requires specific expertise. Reliable taxonomic assignment may be strengthened by the analysis of DNA sequences. Species-specific PCR assays are available for most Fusarium pathogens, but the number of species that infect maize increases the labor and costs required for analysis. In this work, a diagnostic assay for major Fusarium pathogens of maize based on the analysis of melting curves of PCR amplicons was established. Short segments of genes RPB2 and TEF-1α, which have been widely used in molecular taxonomy of Fusarium, were amplified with universal primers in a real-time thermocycler and high-resolution melting (HRM) curves of the products were recorded. Among major Fusarium pathogens of maize ears, F. cerealis, F. culmorum, F. graminearum, F. equiseti, F. poae, F. temperatum, F. tricinctum, and F. verticillioides, all species except for the pair F. culmorum/F. graminearum could be distinguished by HRM analysis of a 304 bp segment of the RPB2 gene. The latter two species could be differentiated by HRM analysis of a 247 bp segment of the TEF-1α gene. The assay was validated with DNA extracted from pure cultures of fungal strains, successfully applied to total DNA extracted from infected maize ears and also to fungal mycelium that was added directly to the PCR master mix (“colony PCR”). HRM analysis thus offers a cost-efficient method suitable for the diagnosis of multiple fungal pathogens.

Fusarium subglutinans is a plant pathogenic fungus infecting cereal grain crops. In 2011, the species was divided in Fusarium temperatumsp. nov. and F. subglutinans sensu stricto. In order to determine the occurrence and significance of F. temperatum and F. subglutinans on maize, a monitoring of maize ears and stalks was carried out in Germany in 2017 and 2018. Species identification was conducted by analysis of the translation elongation factor 1α (TEF-1α) gene. Ninety-four isolates of F. temperatum and eight isolates of F. subglutinans were obtained during two years of monitoring from 60 sampling sites in nine federal states of Germany. Inoculation of maize ears revealed a superior aggressiveness for F. temperatum, followed by Fusarium graminearum, Fusarium verticillioides, and F. subglutinans. On maize stalks, F. graminearum was the most aggressive species while F. temperatum and F. subglutinans caused only small lesions. The optimal temperature for infection of maize ears with F. temperatum was 24 °C and 21 °C for F. subglutinans. All strains of F. temperatum and F. subglutinans were pathogenic on wheat and capable to cause moderate to severe head blight symptoms. The assessment of mycotoxin production of 60 strains of F. temperatum cultivated on rice revealed that all strains produced beauvericin, moniliformin, fusaric acid, and fusaproliferin. The results demonstrate a higher prevalence and aggressiveness of F. temperatum compared to F. subglutinans in German maize cultivation areas.

Fusarium culmorum is a major pathogen of grain crops. Infected plants accumulate deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), or nivalenol (NIV), which are mycotoxins of the trichothecene B group. These toxins are also produced by F. graminearum species complex. New trichothecenes structurally similar to trichothecenes B but lacking the carbonyl group on C-8, designated NX toxins, were recently discovered in atypical isolates of F. graminearum from North America. Only these isolates and a few strains of a yet to be characterized Fusarium species from South Africa are known to produce NX-2 and other NX toxins. Here, we report that among 20 F. culmorum strains isolated from maize, wheat, and oat in Europe and Asia over a period of 70 years, 18 strains produced NX-2 simultaneously with 3-ADON and DON or NIV. Rice cultures of strains producing 3-ADON accumulated NX-2 in amounts corresponding to 2–8% of 3-ADON (1.2–36 mg/kg). A strain producing NIV accumulated NX-2 and NIV at comparable amounts (13.6 and 10.3 mg/kg, respectively). In F. graminearum, producers of NX-2 possess a special variant of cytochrome P450 monooxygenase encoded by TRI1 that is unable to oxidize C-8. In F. culmorum, producers and nonproducers of NX-2 possess identical TRI1; the reason for the production of NX-2 is unknown. Our results indicate that the production of NX-2 simultaneously with trichothecenes B is a common feature of F. culmorum.