Food Safety Research Information Office
Title:Developing Resistance to Aflatoxin through Seed-Based Technologies
Objective:

1. Develop and implement marker-assisted corn breeding strategy. Identify and characterize novel markers associated with aflatoxin-resistance, e.g., resistance-associated proteins (RAPs), in developing and mature kernels through proteomic and genomic comparisons of resistant and susceptible corn genotypes.

2. Identify new sources of corn germplasm and develop new germplasm resistant to fungal infection and aflatoxin contamination with national and international collaboration, using laboratory and field inoculations of corn kernels with tester fungi designed for rapid resistance screening.

3. Evaluate the contribution of novel RAP genes from corn (see Objective 1) for resistance to A. flavus growth and aflatoxin production and use these genes or others to develop transgenic cotton with enhanced resistance to aflatoxin contamination under greenhouse and field conditions. Identify and transfer resistant varieties to cooperating plant breeders for development of varieties for commercial application.

4. Develop rapid, non-destructive hyperspectral imaging methodology to:

    a) measure fungal growth and aflatoxin contamination in corn as a tool for use in enhancement of Homeland Security, and
    b) measure spectral signatures associated with traits for resistance to fungal infection and aflatoxin contamination in corn kernels.

More Info:
Approach:
Resistance to aflatoxin contamination will be enhanced in corn and cottonseed through marker assisted breeding and genetic engineering, respectively. In order to accomplish these goals, complex natural resistance mechanisms in corn kernels will be elucidated in resistant corn inbreds through identification of resistance associated proteins using proteomics and other resistance associated compounds through chemical analysis. Understanding the molecular basis of seed based resistances will lead to identification of biochemical factors correlated with resistance for use in marker assisted breeding and/or when pertinent resistance genes are identified and cloned, for use in enhancement of resistance in crops through genetic engineering. This strategy is especially pertinent to cottonseed, which does not possess practical levels of natural resistance to aflatoxin producing fungi in its germplasm base. Another goal is to assess resistance related biochemical products for their stability of expression in native and transgenic crops under environmental conditions (e.g. drought) known to be conducive to aflatoxin contamination. Also as a part of this project, rapid, non destructive detection methodology based upon hyperspectral imaging will be developed to measure fungal growth and aflatoxin in corn kernels and spectral signatures associated with traits for resistance to fungal infection and aflatoxin contamination in corn kernels, and also to measure physical and biochemical attributes in kernels potentially useful in resistance marker selection.
Funding Source:United States Department of Agriculture (USDA), Agricultural Research Service (ARS)
Type:Appropriated
Start Date:2010
End Date:2015
Project Number:6435-42000-021-00
Accession Number:420817
Institutions:USDA/ARS - Mid South Area
Investigators:Bhatnagar, Deepak
Brown, Robert
Cary, Jeffrey
Rajasekaran, Kanniah
Yu, Jiujiang
Project Reports:2013 Annual Report
2012 Annual Report
2011 Annual Report
Published Journal
Articles USDA
ARS (NP 108):
Co-inoculating of aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus to study fungal invasion, colonization, and competition in maize kernels
Hruska Z, Rajasekaran K, Yao H, Kincaid R, Darlington D, Brown RL, Bhatnagar D, Cleveland TE.
Front Microbiol. 2014 Mar 27. [Epub ahead of print]
Functional characterization of a veA-dependent polyketide synthase gene in Aspergillus flavus necessary for the synthesis of asparasone, a sclerotium-specific pigment
Cary JW, Harris-Coward PY, Ehrlich KC, Di Mavungu JD, Malysheva SV, De Saeger S, Dowd PF, Shantappa S, Martens SL, Calvo AM.
Fungal Genet Biol. 2014 Mar;64:25-35.
Fluorescence Imaging Spectroscopy (FIS) for Comparing Spectra from Corn Ears Naturally and Artificially Infected with Aflatoxin Producing Fungus
Hruska Z, Yao H, Kincaid R, Darlington D, Brown RL, Bhatnagar D, Cleveland TE.
J Food Sci. 2013 Aug;78(8):T1313-20.
Detecting maize inoculated with toxigenic and atoxigenic fungal strains with fluorescence hyperspectral imagery
Yao H, Hruska Z, Kincaid R, Brown RL, Bhatnagar D, Cleveland TE .
Biosystem Eng. 2013 Jun;115(2):125-135.
Evaluation of resistance to aflatoxin contamination in kernels of corn genotypes using a GFP-expressing Aspergillus flavus strain
Rajasekaran K, Sickler CM, Brown RL, Cary JW, Bhatnagar D.
World Mycotoxin J. 2013 May;6(2):151-8.
Evaluation of resistance to aflatoxin contamination in kernels of maize genotypes using a GFP-expressing Aspergillus flavus strain
Rajasekaran K, Sickler CM, Brown RL, Cary JW, Bhatnagar D.
World Mycotoxin J. 2013 May;6(2):151-8.
Identifying cotton (Gossypium hirsutum L.) genes induced in response to Aspergillus flavus infection
Lee S, Rajasekaran K, Ramanarao MV, Bedre R, Bhatnagar D, Baisakh N.
Physiol Mol Plant Pathol. 2012 Oct;80:35-40.
Antifungal activities of Hedychium essential oils and plant extracts against mycotoxigenic fungi
Rajasekaran K, Sakhanokho HF, Tabanca N.
J Crop Improv. 2012 May 30;26:389-96.
Functional and phylogenetic analysis of the Aspergillus ochraceoroseus aflQ (ordA) gene ortholog
Cary JW, Harris-Coward PY, Ehrlich KC, Moore GG, Wei Q, Bhatnagar D.
Mycologia. 2012 Apr 11;104(4):857-64.
The veA gene of the pine needle pathogen Dothistroma septosporum regulates sporulation and secondary metabolism
Chettri P, Calvo AM, Cary JW, Dhingra S, Guo Y, McDougal RL, Bradshaw RE.
Fungal Genet Biol. 2012 Feb;49(2):141-51.
Non-Journal Publications:
Breeding aflatoxin resistant maize lines using recent advances in technologies-a review - (Review Article)
Brown, R.L., Menkir, A., Chen, Z-Y., Bhatnagar, D., Yu, J., Yao, H., Cleveland, T.E. 2013. Breeding aflatoxin resistant maize lines using recent advances in technologies-a review. Food Additives & Contaminants. Part A, (30)8:1382-1391.
Development of maize host resistance to aflatoxigenic fungi - (Book / Chapter)
Brown, R.L., Bhatnagar, D., Cleveland, T.E., Chen, Z.-Y., Menkir, A. 2013. Development of maize host resistance to aflatoxigenic fungi. In: Prof. Mehdi Razzaghi-Abyaneh (Ed.). Aflatoxins - Recent Advances and Future Prospects. ISBN: 978-953-51-0904-4. InTech, Available: http://www.intechopen.com/books/aflatoxins-recent-advances-and-future-prospects/development-of-maize-host-resistance-to-aflatoxigenic-fungi.
Biolistic transformation of cotton zygotic embryo meristem - (Book / Chapter)
Rajasekaran, K. 2013. Biolistic transformation of cotton zygotic embryo meristem. In: Zhang, B-H. (editor). Transgenic Cotton: Methods and Protocols, Methods in Molecular Biology. Humana Press. 958:47-57.
Biolistic transformation of cotton embryogenic cell suspension cultures - (Book / Chapter)
Rajasekaran, K. 2013. Biolistic transformation of cotton embryogenic cell suspension cultures. In: Zhang, B-H. (editor). Transgenic Cotton: Methods and Protocols, Methods in Molecular Biology. Humana Press. 958:59-70.
Utilizing fluorescence hyperspectral imaging to differentiate corn inoculated with toxigenic and atoxigenic fungal strains - (Proceedings)
Yao, H., Hruska, Z., Kincaid, R., Brown, R.L., Bhatnagar, D., Cleveland, T.E. 2012. Utilizing fluorescence hyperspectral imaging to differentiate corn inoculated with toxigenic and atoxigenic fungal strains. In: Proceedings of 2012 SPIE Conference, April 24-25, 2012, Baltimore, Maryland. p 8369-10.
SVM-based feature extraction and classification of aflatoxin contaminated corn using fluorescence hyperspectral data - (Proceedings)
Accepted Publication (09-Apr-12)
Small wonders: peptides for disease control - (Book / Chapter)
Rajasekaran, K., Cary, J.W., Jaynes, J.M., Montesinos, E. 2012. Small wonders: peptides for disease control. American Chemical Society Symposium Series 1095. American Chemical Society, Washington, DC: Oxford University Press, Inc. 472 p.
Preface - (Book / Chapter)
Rajasekaran, K., Cary, J.W., Jaynes, J.M., Montesinos, E. 2012. Preface. In: Rajasekaran, K., Cary, J.W., Jaynes, J.M., Montesinos, E., editors. Small Wonders: Peptides for Disease Control. American Chemical Society Symposium Series. American Chemical Society, Washington, DC: Oxford University Press, Inc. p. ix-xi.
Strategies for controlling plant diseases and mycotoxin contamination using antimicrobial synthetic peptides - (Book / Chapter)
Rajasekaran, K., Cary, J.W., Chlan, C.A., Jaynes, J.M., Bhatnagar, D. 2012.Strategies for controlling plant diseases and mycotoxin contamination using antimicrobial synthetic peptides. In: Rajasekaran, K., Cary, J.W., Jaynes, J.M., Montesinos, E., editors. Small Wonders: Peptides for Disease Control. American Chemical Society Symposium Series. American Chemical Society, Washington, DC: Oxford University Press, Inc. p. 295-315.
Novel synthetic peptides control mycotoxigenic fungi - (Abstract Only)
Accepted Publication (21-Oct-11)
Identification of gene markers in aflatoxin-resistant maize germplasm for marker-assisted breeding - (Book / Chapter)
Brown, R.L., Menkir, A., Chen, Z.-Y., Luo, M., Bhatnagar, D. 2011. Identification of gene markers in aflatoxin-resistant maize germplasm for marker-assisted breeding. In: Guevara-Gonzalez, R.G., editor. Aflatoxins - Biochemistry and Molecular Biology. Rijeka, Croatia: Intech Open Access Publishers. pp 91-106.
Developing resistance to aflatoxin in maize and cottonseed - (Review Article)
Cary, J.W., Rajasekaran, K., Brown, R.L., Luo, M., Chen, Z.-Y., Bhatnagar, D. 2011. Developing resistance to aflatoxin in maize and cottonseed. Toxins. 3:678-696.
Development of narrow-band fluorescence index for the detection of aflatoxin contaminated corn - (Proceedings)
Yao, H., Hruska, Z., Kincaid, R., Ononye, A., Brown, R.L., Bhatnagar, D., Cleveland, T.E. 2011. Development of narrow-band fluorescence index for the detection of aflatoxin contaminated corn. Proceedings of 2011 SPIE Conference, Sensing for Agriculture and Food Quality and Safety III, April 26-27, 2011, Orlando, FL. p.8027-12.
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Government Policy and Regulations
Methodology
Plant Science & Plant Products
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