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| Web URL(s): | http://archive.lib.msu.edu/tic/rpr/1999/72323,%20Ok%20State, Anderson.PDF
Last checked: 04/04/2013 |
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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; Taliaferro, Charles |
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| Monograph Title: | Transformation of Bermudagrass for Improved Fungal Tolerance: USGA Project Annual Report for 1999, 1999. |
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| Publishing Information: | Stillwater, Oklahoma: Oklahoma State University, Department of Plant and Soil Sciences |
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| # of Pages: | 13 |
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| Collation: | 13 pp. |
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| Abstract/Contents: | "Spring dead spot is a major bermudagrass disease in the Southern United States. The causal agent is Ophiosphaerella herpotricha and Ophiosphearella korrea. The most susceptible varieties include many high quality vegetatively progated bermudas. Improvement in resistance in these sexually isolated lines require the development of genetic transformation protocols. The objectives of this work is to first develop an efficient bermudagrass transformation system, second, to isolate genes and/or factors with specific activity against the causal agent of spring dead spot and other fungal diseases, and third, to utilize these agents or genes to increase resistance to fungal diseases. Transformation of the bermudagrass variety Brazos was performed using the Biolistics bombardment with DNA containing an expression cassette that include the bar gene coupled to a constitutive ubiquitin promoter. The gar gene codes for an enzyme that metabolizes the herbicide Liberty TM. After bombardment Liberty was used in the selection media to screen for resistant transgenic tissues. Current efforts to screen transgenic plants using PCR and Southern hybridization techniques identified three more consistent positives (P252, P303, and P319). Of the three positives, P303, showed more intense hybridization possibly indicating incorporation of multiple copies of the bar gene. Furthermore, in an herbicide swipe test P303 showed higher levels of resistance to the herbicide Liberty. Further work to exhaustively screen all 671 transformants is in progress. A microorganism with potent activity against O. herpotricha was recently discovered and characterized. Activity of the antifungal factor was stable in vitro over a six month period. The microorganism was identified to the genus level using GC-FAME and Biolog analysis. Activity was due to the secretion of a potent antifungal factor that was stable under the harshest conditions. Idolation of the factor using preparative SDS PAGE and reverse phase chromatography was successful. The compound was identified through mass spectroscopy and other analytical techniques. Currently we are interested in developing either the microorganism or the antifungal factor as a biocontrol agent. Chitinases are well known enzymes that possess potent antifungal activity. To determine if chitinases play a role in the resistance mechanism against O. herpotricha we inoculated pots with the dungus, grew the plants for more than a month to establish the infection and then transferred the plants to a 15 C temperature, a temperature more conducive to O. herpotricha growth. We developed a novel protocol to isolate individual chitinase isozymes and determine enzyme activity. Isozymes were separated according to their isoelectric point. Infection with O. herpotricha resulted in a doubling of basic chitinases in resistant Midiron, but not Tifgreen. Furthermore, northern blot analysis clearly showed an increase in basic chitinase gene expression at 15 C. This data indicates that basic chitases may have a role in disease resistance in bermuda crown tissues." |
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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", 1999 Turfgrass and Environmental Research Summary [USGA], p. 38-39, 1999, R=72323. R=72323 |
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| Note: | "USGA Project Annual Report for 1999"
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Last checked: 04/04/2013 |
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