title: Stable plastid transformation in lettuce (Lactuca sativa L.).
abstract: Although plastid transformation in higher plants was first demonstrated in the early 1990s it is only recently that the technology is being extended to a broader range of species. To date, the production of fertile transplastomic plants has been reported for tobacco, tomato, petunia, soybean, cotton and Lesquerella fendleri (Brassicaceae). In this study we demonstrate a polyethylene glycol-mediated plastid transformation system for lettuce that generates fertile, homoplasmic, plastid-transformed lines. Transformation was achieved using a vector that targets genes to the trnA/trnI intergenic region of the lettuce plastid genome employing the aadA gene as a selectable marker against spectinomycin. Spectinomycin resistance and heterologous gene transcription were shown in T(1) plants derived from self-pollinated primary regenerants demonstrating transmission of the plastid-encoded transgene to the first seed generation. Crossing with male sterile wild-type lettuce showed that spectinomycin resistance was not transmitted via pollen. Constructs containing the gfp gene showed plastid-based expression of green fluorescent protein. The lettuce plastid could have potential both as a production and a delivery system for edible human therapeutic proteins.
authors.affiliation: Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
authors.forename: Cilia L C
authors.initials: CLC
authors.lastname: Lelivelt
authors.affiliation: Institute of Bioengineering and Agroecology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: McCabe
authors.affiliation: Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK.
authors.forename: Christine A
authors.initials: CA
authors.lastname: Newell
authors.affiliation: Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
authors.forename: C Bastiaan
authors.initials: CB
authors.lastname: deSnoo
authors.affiliation: Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
authors.forename: Kees M P
authors.initials: KMP
authors.lastname: van Dun
authors.affiliation: Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK.|Department of Anatomy, University of Cambridge, Downing Street, CB2 3DY, Cambridge, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Birch-Machin
authors.affiliation: Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK.
authors.forename: John C
authors.initials: JC
authors.lastname: Gray
authors.affiliation: Department of Biochemistry, Trinity College, Dublin 2, Ireland.
authors.forename: Kingston H G
authors.initials: KHG
authors.lastname: Mills
authors.affiliation: Institute of Bioengineering and Agroecology, National University of Ireland, Maynooth, Co. Kildare, Ireland. jackie.nugent@nuim.ie.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Nugent
mesh_terms: D003433:Crosses, Genetic; D004351:Drug Resistance / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D018545:Lactuca / Q000166:cytology* / Q000187:drug effects / Q000235:genetics*; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D018087:Plastids / Q000235:genetics*; D011092:Polyethylene Glycols; D036226:Seedlings / Q000187:drug effects / Q000235:genetics; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D000198:Spectinomycin / Q000494:pharmacology; D014170:Transformation, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics
chemical_list: D010940:Plant Proteins; D011092:Polyethylene Glycols; D000198:Spectinomycin