Progress 06/22/00 to 05/31/04
Outputs
4. What were the most significant accomplishments this past year? D. Progress Report: This report serves to document research conducted under a specific cooperative agreement between ARS and Iowa State University. Additional details of research can be found in the report for the parent project 3620-42000-024-00D, Control of Fusarium verticillioides, Fumonisins and Fusarium Diseases in Corn and Small Grains. This agreement with the Plant Transformation Facility at Iowa State University was set in place to help achieve our goals of developing a gene expression system for corn that would allow proteins that inhibit fungal growth or reduce mycotoxin toxicity to specifically accumulate within the basal maternal tissues of the developing maize kernel, where fungal invasion often begins. A final gene construct was tested for transgenic expression after transformation of corn by the Agrobacterium method. This construct was a needed control linking the
Agrobacterium-mediated transformation work with the previous biolistic-mediated transformations with the basal-kernel-specific promoter. The results showed clearly that the advantages of the Agro transformation method were innate and not the result of a construct modification. Again, the Agrobacterium-mediated transformant resulted in stable gene insertions that were heritable. This is extremely important if the gene expression cassette is to be of value as a biotechnological tool for maize improvement. This work is currently being incorporated into a publication.
Impacts
(N/A)
Publications
- Muhitch, M.J. Distribution of the glutamine synthetase isozyme GSp1 in maize (Zea mays). Journal of Plant Physiology. 2003. v. 160. p. 610-605.
- Muhitch, M.J., Liang, H., Sollenberger, K.G. Transformation efficiencies and expression patterns of a series of truncated GS1-2 promoter/GUS transgenes in maize. Physiologia Plantarum. 2003. v. 118. p. 346-351.
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Progress 10/01/02 to 09/30/03
Outputs
4. What were the most significant accomplishments this past year? D. Progress Report: This report serves to document research conducted under a specific cooperative agreement between ARS and Iowa State University. Additional details of research can be found in the report for the parent project 3620-42000-024-00D, Control of Fusarium verticillioides, Fumonisins and Fusarium Diseases in Corn and Small Grains. This agreement with the Plant Transformation Facility at Iowa State University was set in place to help achieve our goals of developing a gene expression system for corn that would allow proteins that inhibit fungal growth or reduce mycotoxin toxicity to specifically accumulate within the basal maternal tissues of the developing maize kernel, where fungal invasion often begins. Previous transformations with the tissue specific promoter resulted in low transformation efficiencies, so this year, a new approach, using Agrobacterium-mediated transformation was
tested. Analysis of over 40 lines of transgenic corn so transformed resulted in highly improved transformation efficiencies while maintaining gene expression within the maternal tissues of the developing maize kernel. The transformed maize plants also showed strong expression within the vascular tissues of stems, a primary route of fungal growth in infected seedlings. Most importantly, this new transformation method resulted in stable gene insertions which are heritable. This is extremely important if the gene expression cassette is to be of value as a biotechnological tool for maize improvement.
Impacts
(N/A)
Publications
- Muhitch, M.J. Distribution of the glutamine synthetase isozyme GSp1 in maize (Zea mays). Journal of Plant Physiology. 2003. v. 160. p. 610-605.
- Muhitch, M.J., Liang, H., Sollenberger, K.G. Transformation efficiencies and expression patterns of a series of truncated GS1-2 promoter/GUS transgenes in maize. Physiologia Plantarum. 2003. v. 118. p. 346-351.
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