Food Safety Research Information Office
Title:Control of Toxic Endophytic Fungi with Bacterial Endophytes and Regulation of Bacterial Metabolites for Novel Uses in Food Safety
Objective:

1. Determine the potential and genetic mechanisms involved in horizontal gene transfer from Gram-negative pathogenic bacteria to a plant friendly Gram-positive bacterial endophyte Bacillus mojavensis affecting its use as a desirable biocontrol potential.

2. Determine the diversity, evolution, and function of bacterial endophytes in cornfield environments and their impact on the metabolic activity of Fusarium verticillioides, including the production of mycotoxins as well as the detoxification/inactivation of xenobiotic compounds.

3. Determine how the biocontrol organisms can be effectively utilized to prevent the accumulation of the fumonisins; specifically the enhancement of plant defense strategies and production of the antifungal agent surfactin by Bacillus mojavensis.

More Info:
Approach:
Objective 1: We will test to determine if pathogenic Gram-negative bacteria can transfer genetic information via conjugation to Gram-positive endophytic bacteria that could potentially alter an otherwise nontoxic plant friendly bacterium to a potential virulent human pathogen while maintaining its stable endophytic host relationship.
Objective 2: A multilevel approach using molecular genetics, bioinformatics, and in planta analyses will be used to provide data on variation among mycotoxin-producing Fusarium species. Emphasis will be on genes encoding diverse metabolic activities, including xenobiotic detoxification. The hypothesis that such genes were acquired via horizontal gene transfer will be addressed. The approach will evaluate the contribution of these metabolic activities to the general fitness and competitiveness of the fungi.
Objective 3: The same bank of strains of B. mojavensis used in the experimental procedures of Objective 1 will be used here as needed (see Appendix, Table 1), as will the seedling bioassay (Appendix, Figure 6) developed for rating the in planta control of B. mojavensis strains under control conditions. Two cultivars of corn will be used, one rated resistant to Fusarium and the other susceptible. Most of the techniques, inoculations, measurements of disease expressions, endophytism, surface disinfections, microscopic and visual interpretation of diseases have been practiced or created in our laboratory over the years of studying this and other endophytic associations. All analytical analyses for surfactin and fumonisins will be determined as we have done over the past research accomplishments.
Funding Source:United States Department of Agriculture (USDA), Agricultural Research Service (ARS)
Type:Appropriated
Start Date:2011
End Date:2016
Project Number:6612-42000-042-00
Accession Number:421722
Institutions:USDA/ARS - South Atlantic Area
Investigators:Bacon, Charles
Glenn, Anthony (Tony)
Gold, Scott
Hinton, Jr., Arthur
Riley, Ronald
Project Reports:2013 Annual Report
2012 Annual Report
Pending USDA ARS Publications (NP 108):
Published Journal
Articles USDA
ARS (NP 108):
Analyses of black Aspergillus species of peanut and maize for ochratoxins and fumonisins
Palencia ER, Mitchell TR, Snook ME, Glenn AE, Gold S, Hinton DM, Riley RT, Bacon CW.
J Food Prot. 2014 May;77(5):805-13.
Two members of the Ustilago maydis velvet family influence teliospore development and virulence on maize seedlings
Karakkat BB, Gold SE, Covert SF.
Fungal Genet Biol. 2013 Dec;61:111-9.
Metabolites from the entophytic fungus Sporormiella minimoides isolated from Hintonia latiflora
Leyte-Lugo M, Figueroa M, Gonzalez M del C, Glenn AE, Gonzalez-Andrade M, Mata R.
Phytochemistry. 2013 Dec;96:273-8.
Thielavins A, J and K: α-Glucosidase inhibitors from MEXU 27095, an endophytic fungus from Hintonia latiflora
Rivera-Chavez J, González-Andrade M, Gonzalez Mdel C, Glenn AE, Mata R.
Phytochemistry. 2013 Oct;94:198-205.
Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants
Palencia ER, Glenn AE, Hinton DM, Bacon CW.
J Microbiol Methods. 2013 Sep;94(3):381-9.
Plant-symbiotic fungi as chemical engineers: multi-genome analysis of the Clavicipitaceae reveals dynamics of alkaloid loci
Schardl CL, Young CA, Hesse U, Jaromczyk JW, Farman ML, Tudzynski P, Roe BA, Amyotte SG, An Z, Andreeva K, Arnaoudova EG, Bullock CT, Calie P, Charlton ND, Chen L, Fleetwood DJ, Florea S, Guldener U, Harris DR, Haws DC, Jaromczyk J, Johnson RD, Khan AK, Liu J, Liu M, Machado C, Moore N, Nagabhyru P, Oeser B, Pan J, Panaccione DG, Schmid J, Schweri KK, Scott B, Sugawara K, Takach J, Voisey CR, Webb JS, Wiseman J, Zeng Z, Cox M, Dinkins RD, Glenn AE, Gordon A, Hollin W, Leistner E, Leuchtmann A, Li C, Mace W, O'Sullivan DM, Steiner U, Tanaka E, Wilson EV, Yoshida R.
PLoS Genetics. 2013 Feb 28;9(2):e1003323.
The virally encoded killer proteins from Ustilago maydis
Allen A, Islamovic E, Kaur J, Gold S, Shah D, Smith T.
Fungal Biol Reviews. 2013 Jan;26(4):166-73.
(+)-Ascosalitoxin and vermelhotin, a calmodulin inhibitor, from an endophytic fungus isolated from Hintonia latiflora
Leyte-Lugo M, González-Andrade M, González Mdel C, Glenn AE, Cerda-García-Rojas CM, Mata R.
J Nat Prod. 2012 Sep 28;75(9):1571-7.
Deletion of the Ustilago maydis ortholog of the Aspergillus sporulation regulator medA affects mating and virulence through pheromone response
Chacko N, Gold S.
Fungal Genet Biol. 2012 Jun;49(6):426-32.
The secret world of endophytes in perspective
White JF, Bacon CW.
Fungal Ecol. 2012 Jun;5(3):287-8.
Isolation of UmRrm75, a gene involved in dimorphism and virulence of Ustilago maydis
Rodríguez-Kessler M, Baeza-Montañez L, García-Pedrajas MD, Tapia-Moreno A, Gold S, Jiménez-Bremont JF, Ruiz-Herrera J.
Microbiol Res. 2012 May 20;167(5):270-82.
Fungal endophyte diversity in Sarracenia
Glenn A, Bodri MS.
PLoS One. 2012;7(3):e32980.
Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut
Allen A, Islamovic E, Kaur J, Gold S, Shah D, Smith TJ.
Plant Biotechnol J. 2011 Oct;9(8):857-64.
Non-Journal Publications:
The abiotic and biotic plant stress tolerant and beneficial secondary metabolites produced by endophytic Bacillus species - (Book / Chapter)
(04-Apr-14)
Historical accounts of George Washington Carver's contributions to the study of fungi and fungal diseases of plants in the southestern United States - (Abstract Only)
(21-Feb-14)
Nitric oxide detoxification by Fusarium verticillioides involves flavohemoglobins and the denitrification pathway - (Abstract Only)
Baldwin, T.T., Glenn, A.E. 2013. Nitric oxide detoxification by Fusarium verticillioides involves flavohemoglobins and the denitrification pathway [abstract]. Phytopathology 103(Suppl. 2):S2.11. DOI: 10.1094/PHYTO-103-6-S2.11
Genomic and biochemical characterization of cyclic lipopeptides of Bacillus mojavensis RRC101, an antagonist of Fusarium verticillioides - (Abstract Only)
Blacutt, A.A., Gold, S.E., Mitchell, T.R. 2013. Genomic and biochemical characterization of cyclic lipopeptides of Bacillus mojavensis RRC101, an antagonist of Fusarium verticillioides [abstract]. Phtopathology 103(Suppl. 2):S2.16. DOI: 10.1094/PHYTO-103-6-S2.16
Microbial endophytes: future challenges - (Book / Chapter)
Accepted Publication (07-Mar-13)
Additional hosts for Balansia epichloe in tall fescue pastures - (Abstract Only)
Bacon, C.W., Hinton, D.M. 2013. Additional hosts for Balansia epichloe in tall fescue pastures [abstract]. Phytopathology 103(Suppl. 2):S2.11. DOI: 10.1094/PHYTO-103-6-S2.11
Additional hosts for Balansia epichloe in tall fescue pastures - (Abstract Only)
Bacon, C.W., Hinton, D.M. 2013. Additional hosts for Balansia epichloe in tall fescue pastures [abstract]. Phytopathology 103(Suppl. 2):S2.11. DOI: 10.1094/PHYTO-103-6-S2.11
Characterization of strains of Bacillus mojavensis for biocontrol of a maize pathogen - (Abstract Only)
(01-Feb-13)
Detoxification of nitric oxide by flavohemoglobin and the denitrification pathway in the maize pathogen Fusarium verticillioides - (Abstract Only)
Baldwin, T.T., Glenn, A.E. 2013. Detoxification of nitric oxide by flavohemoglobin and the denitrification pathway in the maize pathogen Fusarium verticillioides [abstract]. Fungal Genetics Reports. 60(Suppl):538.
A fungal metallo-beta-lactamase necessary for biotransformation of maize phytoprotectant compounds - (Abstract Only)
Glenn, A.E., Davis, C.B., Snook, M.E., Gold, S.E. 2013. A fungal metallo-beta-lactamase necessary for biotransformation of maize phytoprotectant compounds [abstract]. Fungal Genetics Reports. 60(Suppl):540.
Family disintegration: one fusarium verticillioides beta-lactamase at a time - (Abstract Only)
Gold, S.E., Lin, X., Crenshaw, N.J., Glenn, A.E. 2013. Family disintegration: one fusarium verticillioides beta-lactamase at a time [abstract]. Fungal Genetics Reports 60(Suppl:Abstract #539. p. 253
Rapid deletion plasmid construction methods for protoplast and Agrobacterium based fungal transformation systems - (Book / Chapter)
Garcia-Pedrajas, M.D., Paz, Z., Andrews, D.L., Baeza-Montanez, L., Gold, S.E. 2012. Rapid deletion plasmid construction methods for protoplast and Agrobacterium based fungal transformation systems. In: Gupta, V., Tuohy, M., Ayyachamy, M., Turner, K., O'Donovan, A., editors. Laboratory Protocols in Fungal Biology. New York, NY: Springer-Verlag New York Inc. p. 375-394.
Comparative genomic, phylogenetic, and functional investigation of the xenobiotic metabolizing arylamine N-acetyltransferase enzyme family among fungi - (Abstract Only)
Accepted Publication (15-May-11)
Comparative investigation of the xenobiotic metabolizing arylamine N-acetyltransferase enzyme family among fungi - (Abstract Only)
Accepted Publication (15-May-11)
Assessment of autophagosome formation by transmission electron microscopy - (Book / Chapter)
Nadal, M., Gold, S.E. 2011. Assessment of autophagosome formation by transmission electron microscopy. In: Bolton, M.D., Thomma, B., editors. Methods in Molecular Biology--Plant Fungal Pathogens: Methods and Protocols. New York, NY: Humana Press. p. 481-489.
Allelochemical effects of volatile compounds from Muscodor yucatanensis, an endophytic fungus from Bursera simaruba - (Abstract Only)
Accepted Publication (15-Mar-11)
Hintoniamine, a new calmodulin inhibitor from the endophytic fungal species 39140-2 - (Abstract Only)
Accepted Publication (15-Mar-11)
Nitric Oxide Metabolism and the Role of NO Detoxifying Flavohaemoglobin in Fusarium verticillioides - (Abstract Only)
Accepted Publication (03-Feb-11)
Comparative genomic survey, exon-intron annotation and phylogenetic analysis of NAT-homologous sequences in archaea, protists, fungi, viruses, and invertebrates - (Proceedings)
Glenn, A.E., Karagianni, E.P., Ulndreaj, A., Boukouvala, S. 2010. Comparative genomic survey, exon-intron annotation and phylogenetic analysis of NAT-homologous sequences in archaea, protists, fungi, viruses, and invertebrates. In: Proceedings of the 5th International Workshop on the Arylamine N-Acetyltransferases, September 1-3, 2010, Paris, France. p. 15.
Arylamine N-acetyltransferases in mycotoxigenic and related fungi of agricultural significance - (Proceedings)
Karagianni, E.P., Ormiston, C.B., Fakis, G., Glenn, A.E., Boukouvala, S. 2010. Arylamine N-acetyltransferases in mycotoxigenic and related fungi of agricultural significance. In: Proceedings of the 5th International Workshop on the Arylamine N-Acetyltransferases, September 1-3, 2010, Paris, France. p. 16.
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