BIOCONTROL OF FUMONISIN AND OTHER MYCOTOXINS IN CORN AND TALL FESCUE WITH MICROBIAL ENDOPHYTES
Location: Toxicology and Mycotoxin Research
Project Number: 6612-42000-039-00
Start Date: Apr 25, 2006
End Date: Apr 24, 2011
The purpose of the proposed research is to define and control the relationship between economically important plants and three diverse fungal groups that produce mycotoxins on corn and forage grasses. One group includes fungi belonging to the Liseola section of the Fusarium, represented by Fusarium verticillioides (synonym=F. moniliforme), the second includes the black-spored Aspergillus species, and the third group consists of obligate grass endophytes of the family Clavicipitaceae, species of Neotyphodium Glenn, Bacon & Hanlin. There are four objectives: 1) To decrease the fumonisin mycotoxin content in corn using biocontrol strategies based on the endophytic bacterium Bacillus mojavensis and soil fungi such as Trichoderma koningii; 2) Define the importance of the fumonisins to corn development and plant or fungus survival, as well as its absorption and translocation in corn; 3) To develop the use of novel Neotyphodium fungal endophytes in tall fescue for specific reduction of cattle toxins, and enhanced forage persistence measured by nematode resistance; and 4) To develop an integrated analysis to determine the extent of the mycotoxin ochratoxin A production by isolates of the lesser known but widely prevalent temperate to tropical species Aspergillus niger and A. carbonarius (the Nigri section) that are endophytic to corn kernels, and to determine any co-endophytic interactions with F. verticillioides and/or the fumonisins produced during corn storage.
Objective 1: Pre-harvest control of fumonisin production will be conducted under controlled conditions of soil moisture, soil inoculum content, over time using corn seedlings and plants of three cultivars that will be inoculated with and without our patented bacterial endophyte, Bacillus mojavensis or Trichoderma koningii, each co-inoculated with strains of Fusarium moniliforme. Fumonisin and fusaric acid, and plant performance will be determined under the above conditions. Objective 2: To address the importance of fumonisin on corn development and virulence factor as well as its translocation by plants, several commercial and experimental corn lines will be screened for tolerance or sensitivity to fumonisin, and identified tolerance and sensitive corn lines will be tested for their ability to translocate fumonisin. All plants will be analyzed for fumonisin content using standard chemical procedures and fungal biomass determined by real-time PCR. The mechanism of corn tolerance will be determined with suppression subtraction hybridization of cDNAs and microarray analyses of sensitive and tolerant lines of corn seedlings. Additionally, a specific approach will target corn genes involved in sphingolipid biosynthesis, such as ASC1. Objective 3: Sterile nematode assays, using two or more nematode species, will be used to determine toxic fractions from the tall fescue endophyte-grass association. The nematode assay will be used both with wild-type and novel endophyte-infected tall fescues. Chemical extracts from infected grasses will be used in the bioassay, and all positive extracts will be separated into specific chemical fractions for chemical identification of the toxin responsible for nematode toxicity. Objective 4: Survey both egg and broiler houses (feed, and litter) for the black-spored Aspergillus niger complex, determine if any of these can produce ochratoxin A on one of several media, including corn, and identify positive isolates to species using molecular techniques. Infect corn seedlings with positive A. niger isolates and determine host-parasite interactions, as well as the effects on toxin accumulation in planta, especially when co-infected with fumonisin producing and non-producing strains of F. verticillioides.