Commercially available natural inhibitors of trichothecene production in Fusarium graminearum: A strategy to manage Fusarium head blight of wheat

Abstract

Fusarium Head Blight (FHB) is one of the most important diseases affecting wheat production. The disease causes yield and quality losses and hinders commercialization because of frequent grain contamination with trichothecenes, sesquiterpene mycotoxins that are harmful to human and animals and act as pathogenicity factors for FHB. Although not always effective, application of fungicides is the most common method for FHB management; thus, there is an urgent need to develop new efficient and sustainable tools. We analyzed the inhibitory activity of a group of commercially available naturally occurring compounds on the production of trichothecenes and on the severity of FHB on field-grown wheat spikes inoculated with Fusarium graminearum. The gene TRI5 codifies trichodiene synthase, the enzyme that catalyzes the synthesis of trichodiene, a volatile intermediate to trichothecenes. Our first aim was to obtain a trichothecene non-producing mutant of the pathogen by gene disruption of this gene. Seventeen putative ΔTRI5 mutants produced in the laboratory were characterized for pathogenicity and trichothecene production. The target gene was correctly and efficiently replaced in more than 75% of the transformants obtained, indicating a high efficacy of the approach used for gene disruption. Selected commercially available natural inhibitors -eugenol, apocynin, caffeic acid and propyl gallate-, were used to test their effect on trichodiene production in vitro and in field-grown wheat spikes inoculated with F. graminearum, using a ΔTRI5 mutant as non-trichodiene production control. Fungal volatile organic compounds production was measured by gas chromatography coupled to mass spectrometry. Caffeic acid, apocynin and eugenol produced a significant reduction of trichodiene production in vitro. In field assays, sesquiterpene inhibition was also significant at 2 days post inoculation (dpi), showing that effective inhibition of the trichothecene precursor can be achieved through the action of commercially available naturally occurring compounds. However, at later stages (>7 dpi) the production of trichodiene was similar to that of the F. graminearum-inoculated control. At 21 dpi, FHB severity did not differ between the treatments with inhibitors and the inoculated control. Hence, further experiments are required to identify the most appropriate inhibitors concentrations and formulations to improve their bioavailability and to achieve long lasting protection against F. graminearum infections.