Trichoderma atroviride as a promising biocontrol agent in seed coating for reducing Fusarium damping-off on maize.

AIMS: The objective of this work was to identify a fungal strain showing potential biocontrol abilities against two Fusarium damping-off agents and to test it as a Biological Control Agent (BCA) in maize seed coating under field conditions. METHODS AND RESULTS: A collection of native fungal strains associated with maize in Belgium was screened for antagonistic potential against Fusarium avenaceum and Fusarium culmorum. The strain with highest biocontrol potential was identified as an endophytic Trichoderma atroviride BC0584. In greenhouse, it significantly improves the emergence of seedlings infected by F. avenaceum or F. culmorum pathogens. In most field trials carried out during the season 2017, it significantly increased the emergence rate of infected seedlings compared to untreated seeds. One slurriable powder formulation allows BCA conidia to survive over a 6-month storage period at 4°C. CONCLUSIONS: The fungal BC0584 strain is a promising BCA that could be an alternative to synthetic fungicides. It is adapted to local environmental conditions, is easily and cheaply produced and can be stored in a low-cost formulation. SIGNIFICANCE AND IMPACT OF THE STUDY: In Belgium, this is the first study to use a T. atroviride native strain against Fusarium damping-off on maize crop. Modes of action and required conditions for ensuring high biocontrol activity in the field have still to be investigated.

Screening survey of co-production of fusaric acid, fusarin C, and fumonisins B1, B2 and B3 by Fusarium strains grown in maize grains.

Fusarium species isolated from Belgian maize were screened for their ability to produce fusarin C, fusaric acid, fumonisins B1 (FB1), FB2 and FB3 in maize grains. First, cultivation of Fusarium species in Myro liquid medium allowed overcoming the shortage of the standard of fusarin C on the market. All Fusarium verticillioides produced much higher contents of mycotoxins in Myro compared to Fusarium graminearum or Fusarium venenatum. The optimization of the LC-MS/MS method resulted in low limits of detection and quantification for fusarin C, fusaric acid, FB1, FB2 and FB3 determination in maize grains. Its application for screening the potential toxin production ability evidenced that the concentrations of the analytes were significantly increased at various levels when F. verticillioides strains were cultivated in maize grains and reached 441 mg kg(-1) for fusaric acid, 74 mg kg(-1) for fusarin C, 1,301 mg kg(-1) for FB1, 367 mg kg(-1) for FB2 and 753 mg kg(-1) for FB3.

First Report of Fusarium temperatum Causing Seedling Blight and Stalk Rot on Maize in Spain.

In Europe, several diseases of maize (Zea mays L.) including seedling blight and stalk rot are caused by different Fusarium species, mainly Fusarium graminearum, F. verticillioides, F. subglutinans, and F. proliferatum (3). In recent years, these Fusarium spp. have received significant attention not only because of their impact on yield and grain quality, but also for their association with mycotoxin contamination of maize kernels (1,4). From October 2011 to October 2012, surveys were conducted in a maize plantation located in Galicia (northwest Spain). In each sampling, 100 kernels and 10 maize stalks were collected from plants exhibiting symptoms of ear and stalk rot. Dried kernels and small stalk pieces (1 to 2 cm near the nodes) were placed onto potato dextrose agar medium and incubated in the dark for 7 days. Fungal colonies displaying morphological characteristics of Fusarium spp. (2) were subcultured as single conidia onto SNA (Spezieller Nahrstoffarmer agar) (2) and identified by morphological characteristics, as well as by DNA sequence analysis. A large number of Fusarium species (F. verticillioides, F. subglutinans, F. graminearum, and F. avenaceum) (1,2) were identified. These Fusarium species often cause ear and stalk rot on maize. In addition, a new species, F. temperatum, recently described in Belgium (3), was also identified. F. temperatum is within the Gibberella fujikuroi species complex and is morphologically and phylogenetically closely related to F. subglutinans (2,3). Similar to previous studies (3), our isolates were characterized based on the presence of white cottony mycelium, becoming pinkish white. Conidiophores were erect, branched, and terminating in 1 to 3 phialides. Microconidia were abundant, hyaline, 0 to 2 septa; ellipsoidal to oval, produced singly or in false heads, and on monophialides, intercalary phialides, and polyphialides. Microconidia were not produced in chains. No chlamydospores were observed (3). Macroconidia in carnation leaf agar medium (2) were hyaline, 3 to 6 septate, mostly 4, falcate, with a distinct foot-like basal cell (2,3). DNA was amplified with primers ITS1/ITS4 and EF1/EF2 (3). Partial sequences of gene EF-1α showed 100% homology with F. temperatum (3) (GenBank Accession Nos. HM067687 and HM067688). DNA sequences of EF-1α gene and ITS region obtained were deposited in GenBank (KC179824, KC179825, KC179826, and KC179827). Pathogenicity of one representative isolate was confirmed using a soil inoculation method adapted from Scauflaire et al., 2012 (4). F. temperatum isolate was cultured on sterile wheat grains. Colonized wheat grains (10 g) were mixed with sterilized sand in 10 cm diameter pots. Ten kernels per pot were surface disinfected in 2% sodium hypochlorite for 10 min, rinsed with sterilized water, drained (4), placed on the soil surface, and covered with a 2 cm layer of sterilized sand. Five pots were inoculated and five uninoculated controls were included. Pots were maintained at 22 to 24°C and 80% humidity for 30 days. Seedling malformations, chlorosis, shoot reduction, and stalk rot were observed on maize growing in inoculated soil and not from controls. F. temperatum was reisolated from the inoculated seedlings but not from the controls. References: (1) B. J. Bush et al. Phytopathology 94:88, 2003. (2) J. F. Leslie et al. The Fusarium Laboratory Manual, page 388. Blackwell Publishing, 2006. (3) J. Scauflaire et al. Mycologia 103:586, 2011. (4) J. Scauflaire et al. Eur. J. Plant Pathol. 133:911, 2012.