Full TGIF Record # 79890
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Web URL(s):https://turf.rutgers.edu/research/abstracts/symposium2002.pdf#page=49
    Last checked: 02/06/2017
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Publication Type:
i
Report
Author(s):Pitcher, Lynne H.; Zilinskas, Barbara A.
Author Affiliation:Department of Plant Biology and Pathology, Rutgers University
Title:Agrobacterium tumefaciens-mediated transformation of turfgrasses using the selectable marker, phosphomannose isomerase
Section:Poster presentations
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Meeting Info.:Cook College, Rutgers, NJ: January 10-11, 2002
Source:Proceedings of the Eleventh Annual Rutgers TurfgrassSymposium. 2002, p. 48-49.
Publishing Information:New Brunswick, NJ: Center for Turfgrass Science, Cook College, Rutgers, The State University of New Jersey
# of Pages:2
Keywords:TIC Keywords: Agrobacterium tumefaciens; Disease resistance; Evaluations; Genetic analysis; Genetic markers; Isomerases; Transgenic turfgrasses
Abstract/Contents:"Genetically modified crops continue to face controversy and opposition, one reason for which is the use of bacterial antibiotic resistance genes as selectable markers and the still debated concern over the effect of these markers on the environment and human health. The combination of E. coli. phosphomannose isomerase (PMI) as a genetic marker with mannose as a selection agent has been suggested as being more environmentally friendly and socially acceptable, as well as possibily less injurious to the plant tissue to be transformed. A mannose selection system has been described for sugarbeet (Joersbo et al., 1998 and 1999) and maize (Wang et al., 2000) wherein the expression of bacterial PMI allowed the transformed plant cells to use mannose as a source of carbohydrate that would otherwise not be metabolized. Here we report the transformation of three turfgrass species using the mannose selection system. Plasmid pNOV2804 (provided by Syngenta Seeds AG) contains the PMI structural gene (manA) of E. coli which converts mannose-6-P to fructose-6-P. This plasmid was modified to place the expression of PMI under the control of the strong, constitutive Ubi-1 promoter from maize; the resultant plasmid pUBIMAN1 was transferred into Agrobacterium tumefaciens strain Gv3101. Callus was derived from mature seeds of velvet bentgrass cv SR7200, creeping bentgrass cv Crenshaw, and tall fescue cv Plantation and Genesis. After co-cultivation of callus with Agrobacterium containing pUBIMAN, transformed callus was selected by its ability to grow on mannose. Callus, thus selected, was allowed to proliferate on mannose and then tested for increased PMI activity (relative to non-transformed callus) by enzymatic assay (Gracy and Noltmann, 1968). Analysis to date shows that there is much greater potential for this technique with bentgrasses than with tall fescue inasmuch as bentgrasses have a much lower basal PMI activity than does tall fescue, allowing for more certainty in selecting transformed tissue. Calli that proliferated on mannose and had higher PMI activity than non-transformed controls were placed on regeneration media; green shoots were allowed to grow in tissue culture before being transferred to soil. DNA from putative transformants will be subjected to PCR analysis and further confirmation will be by Southern analysis of DNA extracted from the resultant plants."
Language:English
References:4
ASA/CSSA/SSSA Citation (Crop Science-Like - may be incomplete):
Pitcher, L. H., and B. A. Zilinskas. 2002. Agrobacterium tumefaciens-mediated transformation of turfgrasses using the selectable marker, phosphomannose isomerase. Proc. Annu. Rutgers Turfgrass Symp. p. 48-49.
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https://turf.rutgers.edu/research/abstracts/symposium2002.pdf#page=49
    Last checked: 02/06/2017
    Requires: PDF Reader
    Notes: Item is within a single large file
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MSU catalog number: SB 433 .R88
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