Transposition-based plant transformation.
Agrobacterium T-DNAs were used to deliver transposable Dissociation (Ds) elements into the nuclei of potato (Solanum tuberosum) cells. A double-selection system was applied to enrich for plants that only contained a transposed Ds element. This system consisted of a positive selection for the neomycin phosphotransferase (nptII) gene positioned within Ds followed by a negative selection against stable integration of the cytosine deaminase (codA) gene-containing T-DNA. Sixteen of 29 transgenic plants were found to contain a transposed element while lacking any superfluous T-DNA sequences. The occurrence of this genotype indicates that Ds elements can transpose from relatively short extrachromosomal DNA molecules into the plant genome. The frequency of single-copy Ds transformation was determined at 0.3%, which is only about 2.5-fold lower than the potato transformation frequency for backbone-free and single-copy T-DNAs. Because of the generally high expression levels of genes positioned within transposed elements, the new transformation method may find broad applicability to crops that are accessible to Agrobacterium T-DNA transfer.
Yan
Hua
H
Simplot Plant Sciences, J.R. Simplot Company, Boise, Idaho 83706, USA.
Rommens
Caius M
CM
D004251:DNA Transposable Elements / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005818:Genetic Engineering / Q000379:methods*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012231:Rhizobium / Q000378:metabolism; D011198:Solanum tuberosum / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics*
D004251:DNA Transposable Elements; D018744:DNA, Plant; D010940:Plant Proteins