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| Web URL(s): | http://archive.lib.msu.edu/tic/rpr/1998/61853,%20OK%20State, Anderson.PDF
Last checked: 10/21/2013
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| Publication Type:
| Report |
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| Material Type: | Manuscript |
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| Monographic Author(s): | Anderson, Michael P.; Guenzi, Arron C.; Taliaferro, Charles M.; Anderson, Jeffrey A. |
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| Author Affiliation: | Anderson, M., Guenzi, and Taliaferro: Principle Investigator, Department of Plant & Soil Sciences; and Anderson, J.: Principle Investigator, Dep. of Horticulture & Landscape Architecture, Oklahoma State University, Stillwater, Oklahoma |
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| Monograph Title: | Transformation of Bermudagrass for Improved Fungal Resistance: [1998 Annual Research Report] , 1998. |
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| Publishing Information: | Stillwater, Oklahoma: Oklahoma State University |
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| # of Pages: | 16 |
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| Collation: | [2], 14 pp. |
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| Abstract/Contents: | "A major disease commonly known as spring dead spot (SDS) causes significant economic damage to bermudagrass in the Southeastern United States. The causal agent for SDS throughout most of the United States is Ophiosphaerella herpotricha and Ophioshpearella korrea. Both fungal species are very active in the fall and early spring when the temperatures are cool and moisture is plentiful. Infected areas appear as regular circular patches of dead and diseased turf that generally occurs in more mature stands of bermudagrass. The long-term goal of this project is to increase resistance in bermudagrass turf varieties to SDS through gene transformation technology. To accomplish this objective we have three main objectives: 1) to develop an efficient and reliable transformation system for bermudagrass species, 2) to identify and isolate SDS inhibitory factors, and 3) to transform bermudagrass with the genes that code for these anti-SDS factors. This report describes the current progress and results for the development of the transformation system and the isolation and characterization of antifungal actors during FY1998. Bermudagrass Transformation: The use of high velocity microprojectiles (BiolisticsTM) to deliver recombinant DNA into intact plant cells has been successfully utilized to transform many grass species, and is considered the method of choice for most grass species. Immature inflorescences of the bermudagrass cultivar 'Brazos' were used to induce the formation of embryogenic callus tissue. Brazos was chosen for this experiment because it had previously demonstrated superior growth and plant regeneration potential in tissue culture. Tissue was transformed with plasmid containing chimeric genes of interests, the bar and uidA genes, under the control of ubiquitin promoters. The bar and uidA genes serve as a selectable marker and reporter gene, respectively. The GUS enzyme, coded for by uidA, can be assayed by accumulation of fluorogenic products by providing the enzyme substrate. PAT detoxifies bialaphos, the active ingredient in the herbicide 'Liberty' in the selective media, thereby allowing transgenic cells and plants to continue to grow. Six hundred and seventy one putative transgenic plants have been recovered from this experiment. We are currently evaluating these putative transformants with PCR to determine if they contain the bar gene (Figure 1). PCR positive plants will be characterized by Southern analysis and enzyme assays for phosphinothricin acetyl transferase during FY 1999. Anti-SDS Proteins: Living organisms to protect themselves from pathogens, or to give them a competitive advantage for nutrients, produce many antimicrobial compounds. They range from the small molecular weight antibiotics and secondary metabolites to the larger macromolecular proteins and assorted polypeptides. Recently we discovered a bacterium that was strongly and persistently inhibitory towards O. herpotricha. The bacterium was identified to the genus taxonomic level with confidence by a GC-FAME and BIOLOG technology. This bacterium secreted many proteins into the extracellular matrix. Dialysis of the extracellular excretions suggested that the antifungal factor was a protein. Purification of the antifungal proteins on anion exchange, hydroxyapatite, and Mono Q chromatography resulted in the isolation of a 36 kD protein that is most likely expressed as multiple isoforms (Figure 2). Analysis of the purification results suggested that there are at least two distinct antifungal factors antagonistic against O. herpotricha secreted by the bacterium. Experiments are in preparation to identify, sequence, and characterize the 36 kD protein." |
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| Language: | English |
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| References: | 0 |
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| See Also: | See also related summary article "Transformation of bermudagrass for improved fungal resistance" 1998 Turfgrass and Environmental Research Summary [USGA], 1998, p. 47, R=61853. R=61853 |
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| Note: | Also appears as pp. 232-247 in the USGA Turfgrass Research Committee Reporting Binders for 1998.
"November 2, 1998"
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