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| Web URL(s): | http://archive.lib.msu.edu/tic/rpr/1992/Turfgrass/Rutgers-Day.PDF
Last checked: 08/22/2019
Requires: PDF Reader |
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| Publication Type:
| Report |
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| Material Type: | Manuscript |
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| Monographic Author(s): | Day, Peter; Funk, Reed |
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| Author Affiliation: | Day: AgBiotech Center; and Funk: Department of Crop Science, Rutgers |
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| Monograph Title: | Endophytes of Turfgrasses: New Tools and Approaches: [1992 Annual Research Report], 1992. |
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| Publishing Information: | [New Brunswick, New Jersey]: [Rutgers University] |
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| # of Pages: | 18 |
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| Collation: | [18] pp. |
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| Abstract/Contents: | "This project was proposed and initiated by Dr. Peter Day, AgBiotech Center, and Dr. Reed Funk, Department of Crop Science, Rutgers. Our program goals are: (a) to produce a germplasm collection of fungal endophyte-infected grasses concentrating on Poa and Agrostis species; (b) to produce a collection of unifungal endophyte cultures for classical and molecular analysis; (c) to extend the host range of endophytes to grasses where they have not been found; (d) to produce endophyte-specific DNA probes; (e) to use the probes to characterize endophyte variability and produce RFLP maps for taxonomy; (f) to develop turfgrass tissue culture regeneration and transformation systems for the incorporation of agronomically important genes into turfgrass. After extensive screening of collections of Poa and Agrostis species throughout the U.S.A. and Europe (in collaboration with Dr. Jim White, Auburn University, Alabama), we have obtained a limited number of fungal endophyte-infected grasses in these two genera. However, as of now we did not find fungal endophytes in Kentucky bluegrass (Poa pratensis) and Creeping bentgrass (Agrostis palustris). Therefore, we are currently developing an in vitro culture system for extending the host range of endophyte to Kentucky bluegrass and Creeping bentgrass. To date, a collection of unifungal endophyte cultures has been established and contains representative isolates from a wide variety of turfgrass genera. We are using mycelial macerates of these endophyte isolates to inoculate embryogenic callus lines of Poa pratensis and Agrostis palustris to obtain novel combinations of endophyte infected grasses. We have inoculated several embryogenic callus lines of creeping bentgrass with endophytes and successfully regenerated plants. Regenerating grasses were planted in the soil and are currently being examined for the presence of endophyte in the leaf bases. Several are positive. We produced fungal endophyte-specific DNA probes by the polymerase chain reaction (PCR) and diagnostic fingerprints of DNA sequences generated by randomly amplified polymorphic DNA (RAPD) PCR method. RAPD markers obtained with four ten base random DNA primers were successfully used to discriminate among individual endophyte isolates. A phylogenetic tree of endophyte isolates was generated using RAPD markers for Kentucky bluegrass and Creeping bentgrass. The embryogenic callus lines established provide alternative tissues for creating endophyte containing grasses and target callus lines with high regeneration potentials have been established with several varieties of Kentucky bluegrass and Creeping bentgrass. We are using E. coli β-glucuronidase as a reporter gene to assay transient expression of an added gene in grass cells after bombardment. Various parameters were optimized to improve transient expression in cultured bentgrass to high levels. We are currently selecting for stably transformed turfgrass cell lines which express a bar gene conferring resistance to the herbicide bialaphos (BastaTM). For identification of putative transformants, we used a well known tissue culture model system, Black Mexican Sweetcorn (BMS) suspension cells, to obtain BMS clones resistant to bialaphos through particle bombardment and with these clones, we developed a PCR assay for early identification of transgenic turfgrass plants. Regenerants from putative transformed callus will be confirmed by Southern blot hybridization and phosphinothricin acetyltransferase (PAT) assay. Other single gene traits of interest confer insect resistance, virus resistance, and resistance to fungal and bacterial pathogens." |
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| Language: | English |
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| References: | 3 |
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| See Also: | See also related leaflet, Endophytes of Turfgrasses: New Tools and Approaches, 1990, R=306587. R=306587 |
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| Note: | "November 1992"
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| ASA/CSSA/SSSA Citation (Crop Science-Like - may be incomplete):
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Last checked: 08/22/2019
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