The preformed antimicrobial compounds produced by maize, 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3-one and its desmethoxy derivative 2,4-dihydroxy-2H-1,4-benzoxazin-3-one, are highly reactive benzoxazinoids that quickly degrade to the antimicrobials 6-methoxy-2-benzoxazolinone (MBOA) and 2-benzoxazolinone (BOA), respectively. Fusarium verticillioides ؍( F. moniliforme) is highly tolerant to MBOA and BOA and can actively transform these compounds to nontoxic metabolites. Eleven of 29 Fusarium species had some level of tolerance to MBOA and BOA; the most tolerant, in decreasing order, were F. verticillioides, F. subglutinans, F. cerealis ؍( F. crookwellense), and F. graminearum. The difference in tolerance among species was due to their ability to detoxify the antimicrobials. The limited number of species having tolerance suggested the potential utility of these compounds as biologically active agents for inclusion within a semiselective isolation medium. By replacing the pentachloronitrobenzene in Nash-Snyder medium with 1.0 mg of BOA per ml, we developed a medium that resulted in superior frequencies of isolation of F. verticillioides from corn while effectively suppressing competing fungi. Since the BOA medium provided consistent, quantitative results with reduced in vitro and taxonomic efforts, it should prove useful for surveys of F. verticillioides infection in field samples.
Kernels of corn, Zea mays, were inoculated with Fusarium moniliforme to analyze seedling growth and development during endophytic, symptomless infection. In planta F. moniliforme distribution and seedling growth, expressed as shoot diameter, plant height, leaf length, and dry weight, were examined weekly for 28 days after planting. Even though no visible disease symptoms developed, F. moniliforme was isolated from most segments taken from seedlings grown from inoculated, but not noninoculated, kernels from the earliest to the latest sampling. F. moniliforme did not alter the rate or percentage of kernel germination, but seedlings grown from inoculated kernels had suppressed shoot diameter, plant height, leaf length, and plant weight 7 days after planting. However, seedling growth from inoculated kernels was similar to or greater than that from noninoculated kernels at 28 days. Histological modifications in seedlings grown from inoculated kernels included accelerated lignin deposition in shoots and modified chloroplast orientation in leaves. In summary, gross morphology and histology were altered in corn seedlings during symptomless, endophytic infection by F. moniliforme.
Fusarium verticillioides is a very important genus from the aspects of plant disease, cereal production, and food safety. A major concern of this species is its mycotoxins, which are harmful to humans and animals ingesting Fusariumcontaminated food or feed products. The fungus exists as a symptomless intercellular endophyte in both field and sweet maize, but its role during the symptomless state of infection is ambiguous. Most strains produce the fumonisin in large quantities during the preharvest and postharvest periods of maize production, even during the symptomless state. The dual nature of F. verticillioides as both pathogen and a symptomless endophyte indicates a complex relationship with maize. Interactive biotic factors such as plant genetics, along with abiotic factors, may alter the required balanced relationships, resulting in a weakened plant and changing the relationship into a disease, during which mycotoxins are produced. Consequently, the development of appropriate control measures for the virulent state is expected to be difficult. Two biocontrol agents and approaches are also reviewed, along that offering some pre-and postharvest biological control measures designed to reduce maize contamination by F. verticillioides and the fumonisin mycotoxins.
Fusarium moniliforme Sheldon, a biological species of the mating populations within the (italic)Gibberella fujikuroi species complex, i.e., population A [= G. moniliformis (Sheld.) Wineland], is an example of a facultative fungal endophyte. During the biotrophic endophytic association with maize, as well as during saprophytic growth, F. moniliforme produces the fumonisins. The fungus is transmitted vertically and horizontally to the next generation of plants via clonal infection of seeds and plant debris. Horizontal infection is the manner by which this fungus is spread contagiously and through which infection occurs from the outside that can be reduced by application of certain fungicides. The endophytic phase is vertically transmitted. This type infection is important because it is not controlled by seed applications of fungicides, and it remains the reservoir from which infection and toxin biosynthesis takes place in each generation of plants. Thus, vertical transmission of this fungus is just as important as horizontal transmission. A biological control system using an endophytic bacterium, Bacillus subtilis, has been developed that shows great promise for reducing mycotoxin accumulation during the endophytic (vertical transmission) growth phase. Because this bacterium occupies the identical ecological niche within the plant, it is considered an ecological homologue to F. moniliforme, and the inhibitory mechanism, regardless of the mode of action, operates on the competitive exclusion principle. In addition to this bacterium, an isolate of a species of the fungus Trichoderma shows promise in the postharvest control of the growth and toxin accumulation from F. moniliforme on corn in storage.
Biocontrol activity against Fusarium moniliforme was analyzed for a Trichoderma viride strain isolated from root segments of corn plants grown in Piedmont Georgia. The isolate suppressed radial extension of F. moniliforme colonies during cocultivation on potato dextrose agar and fumonisin B1 (FB1) production during incubation of both fungi on corn kernels. T. viride decreased radial extension of F. moniliforme by 46% after 6 days and by 90% after 14 days. Furthermore, the colony diameter of F. moniliforme was less at 14 days than at 5 days, suggesting that F. moniliforme mycelia were undergoing lysis. FB1 production by F. moniliforme on corn kernels decreased by 85% when both organisms were inoculated the same day onto corn kernels and by 72% when inoculation of T. viride was delayed by 7 days after F. moniliforme inoculation. These results are the first to demonstrate that T. viride can suppress FB1 production by F. moniliforme, thereby functioning to control mycotoxin production. Thus, this isolate may be useful in biological control to inhibit F. moniliforme growth as a preharvest agent to prevent disease during plant development and/or as a postharvest agent during seed storage to suppress FB1 accumulation when kernels are dried inadequately.
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