Stable plastid transformation in lettuce (Lactuca sativa L.).
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.
Lelivelt
Cilia L C
CLC
Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
McCabe
Matthew S
MS
Institute of Bioengineering and Agroecology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Newell
Christine A
CA
Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK.
deSnoo
C Bastiaan
CB
Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
van Dun
Kees M P
KMP
Rijk Zwaan Breeding B.V., 1e Kruisweg 9, 4793 RS, Fijnaart, The Netherlands.
Birch-Machin
Ian
I
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.
Gray
John C
JC
Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK.
Mills
Kingston H G
KHG
Department of Biochemistry, Trinity College, Dublin 2, Ireland.
Nugent
Jacqueline M
JM
Institute of Bioengineering and Agroecology, National University of Ireland, Maynooth, Co. Kildare, Ireland. jackie.nugent@nuim.ie.
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
D010940:Plant Proteins; D011092:Polyethylene Glycols; D000198:Spectinomycin