Progress 05/15/04 to 05/14/08
Outputs
OUTPUTS: Tomato plants (Lycopersicon esculentum Miller 'Money Maker' transformed with the oxalate oxidase gene (pVSPB OxO) and the PSE antimicrobial peptide gene (PSE39) were more resistant to Ralstonia solanacearum race 1 than non transformed plants. both transformed and non transformecd plants were grown in Magenta GA7 vessel in a soil less medium supplemented with Gamborg's B-5 basal medium with minimal organics (pH 5.8) in a 24 h light photoperiod (25 umol m-1) at 25C. Plant were inoculated with 100,000,00018 h old bacterial suspension. PARTICIPANTS: The project was used to trained graduate students in agricultural biotechnology. Several graduate and undergraduate students were involved in this research under research assistant Tingting Chen. These students developed skills in tissue culture, transformation technologies, and in plant pathology. TARGET AUDIENCES: Results of this work was presented at seminars, presentations at the Southern Nurserymen's Association and at the annual project directors meetings that were held in conjuction with the American Phytopathological Society Annual meetings. PARTICIPANTS: Six graduate students worked on this project, 1 master's student received her master's degree by doing her theses research on the project. TARGET AUDIENCES: The antimicrobial peptided genes evaluated in this project did not increase disease resistance to Pseudomonas solanacearum or to a fungal plant pathogen (Botritis sp.) but increased plant tolerance to abiotic stresses. Four transformed plants were saved and they are being used in other research projects. PROJECT MODIFICATIONS: Initially, a post-doctoral scientist was to be hired to do the transformation work in two years. Since it was not possible to hire a post-doctoral scientist, we recruited graduate students to perform this work. Because of those reasons the project was extended for two additional years.
Impacts
Six graduate students were trained in biotechnology; one student completed her master's thesis research; regenerated plants are being used in other research projects; tomato plants transformed with antimicrobial peptides resulted in increased resistance to environmental stress
Publications
- No publications reported this period
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: Tomato plants (Lycopersicon esculentum Miller 'Money Maker' transformed with the oxalate oxidase gene (pVSPB OxO) and the PSE antimicrobial peptide gene (PSE39) were more resistant to Ralstonia solanacearum race 1 than non transformed plants. both transformed and non transformecd plants were grown in Magenta GA7 vessel in a soil less medium supplemented with Gamborg's B-5 basal medium with minimal organics (pH 5.8) in a 24 h light photoperiod (25 umol m-1) at 25C. Plant were inoculated with 100,000,00018 h old bacterial suspension.
PARTICIPANTS: The project was used to trained graduate students in agricultural biotechnology. Several graduate and undergraduate students were involved in this research under research assistant Tingting Chen. These students developed skills in tissue culture, transformation technologies, and in plant pathology.
TARGET AUDIENCES: Results of this work was presented at seminars, presentations at the Southern Nurserymen's Association and at the annual project directors meetings that were held in conjuction with the American Phytopathological Society Annual meetings.
Impacts
The objective of this project was to evaluate the effect of the oxalate oxidase gene and the PSE antimicrobial peptide gene in transformed plants inoculated with a bacterial plant pathogen. Plantlets were regenerated from various tissues and transformed using a bacterial vector. Once transformed tissues developed into plantlets, they were inoculated in vitro.
Publications
- Sauve, R.J. and Zhou, S. 2007. Effects of Ralsonia solanacearum race 1 on tomato plants transformed with the oxalate oxidase gene. Project Directors July 30 meeting, APS Annual meeting, San Diego, CA.
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Progress 01/01/06 to 12/31/06
Outputs
The goals of this research project were: 1) to develop an efficient transformation system for tomato; 2) to transform tomato with the wheat oxalate oxidase gene; and 3) to later test transgenic plants against different pathogens, esp. Ralstonia solanacearum. Tomato plants were regenerated from hypocotyl and cotyledon explants obtained from ten-day-old seedlings. Explants were plated on Murashige and Skoog supplemented with 0.1 mg/L of IAA with 1 mg/L of zeatin. For genetic transformation, calli generated from cotyledon and hypocotyls were innoculated with the bacterium Agrobacterium tumefaciens EHA105 harboring the gene vectors for the Oxalate oxidase gene that confers resistance to plant pathogenic bacteria and fungil. Transgenic shoots were regenerated on selection media containing glufosinate ammonium. PCR tests and sequence analysis were used to screen putatively transformed plants.
Impacts
Economic losses that could result from Ralstonia, not including field crops such as the potato, are estimated to in the tens of millions to Geranium producers alone. This project examines the effectiveness of antibacterial peptide genes in transformed tomato for resistance to bacterial diseases.
Publications
- No publications reported this period
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Progress 01/01/05 to 12/31/05
Outputs
Tomato plants(Lycopersicon esculentum Miller) Money-Maker cultivar were regenerated from hypocotyl and cotyledon explants obtained from ten-day-old seedlings. Explants were plated on Murashige and Skoog (MS) supplemented with 0.1 mg/L of IAA with 1 mg/L of zeatin. For genetic transformation, calli generated from cotyledon and hypocotyls were infected with a bacterium (Agrobacterium tumefaciens EHA105) harboring the gene vectors for the Oxalate oxidase gene (pVSPB OxO) and PSE antimicrobial peptide gene (PSE39) that confer resistance to bacterial and fungal plant pathogens. Transgenic shoots were regenerated on selection media containing kanamycin or glufosinate ammonium. PCR tests were used to screen putatively transformed plants. The tomato was used as a model plant to evaluate the effects of these genes on disease resistance. Plants are now being propagated for screening for resistance to bacterial plant pathogens.
Impacts
Economic losses that could result from Ralstonia, not including field crops such as the potato, are estimated to in the tens of millions to Geranium producers alone. This project examines the effectiveness of antibacterial peptide genes in transformed tomato for resistance to bacterial diseases.
Publications
- No publications reported this period
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Progress 01/01/04 to 12/31/04
Outputs
The regeneration system for tomato has been optimized and plant tissue is being transformed with Agrobacterium tumefaciens. We have encountered difficulties in cleaning up transformed tissue. Different antibiotic combinations are presently being evaluated. Tomato plants were regenerated from hypocotyls and cotyledon of 10-days-old seedlings. Murashige and Skoog medium supplemented with 5 different combinations of auxins and cytokinins were used. After four weeks of incubation shoots were formed from cotyledons on 3 combinations of cytokinins and auxinx and from hypocotyls on four different combinations of cytokinins. The combination of 0.1 mg/L of IAA with 1 mg/L of Zeatin produced the best shoot quality and in the most number from cotyledon explants.
Impacts
Development of tomato cultivars resistant to bacterial diseases.
Publications
- No publications reported this period
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