A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques.
Two barley transformation systems, Agrobacterium-mediated and particle bombardment, were compared in terms of transformation efficiency, transgene copy number, expression, inheritance and physical structure of the transgenic loci using fluorescence in situ hybridisation (FISH). The efficiency of Agrobacterium-mediated transformation was double that obtained with particle bombardment. While 100% of the Agrobacterium-derived lines integrated between one and three copies of the transgene, 60% of the transgenic lines derived by particle bombardment integrated more than eight copies of the transgene. In most of the Agrobacterium-derived lines, the integrated T-DNA was stable and inherited as a simple Mendelian trait. Transgene silencing was frequently observed in the T1 populations of the bombardment-derived lines. The FISH technique was able to reveal additional details of the transgene integration site. For the efficient production of transgenic barley plants, with stable transgene expression and reduced silencing, the Agrobacterium-mediated method appears to offer significant advantages over particle bombardment.
Travella
S
S
John Innes Centre, Norwich Research Park, Colney, Norwich, NR4 7UH, UK.
Ross
S M
SM
Harden
J
J
Everett
C
C
Snape
J W
JW
Harwood
W A
WA
D000383:Agriculture / Q000379:methods*; D019470:Biolistics; D002875:Chromosomes / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018628:Gene Dosage; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D018745:Genome, Plant; D001467:Hordeum / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D012231:Rhizobium / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics; D019076:Transgenes / Q000235:genetics
D018744:DNA, Plant