Chromosomal localization of cotransformed transgenes in the hexaploid cultivated oat Avena sativa L. using fluorescence in situ hybridization.
Fluorescence in situ hybridization (FISH) was used to localize two transgenes (gus and bar), carried on plasmids pACT-1F and pUBA, respectively, on mitotic metaphase squashes of T1 plants of the cultivated hexaploid oat Avena sativa L. cotransformed by microprojectile bombardment of embryogenic callus. Among the eight progeny analysed by FISH in each of two lines, we detected plants null, hetero- and homozygous for the two genes in one line, and plants null and heterozygous for the two genes in the other line. Our results demonstrated that in the two independent transformation events, the gus and bar genes had inserted in the same position relative to each other. In each transformation event, the insertions occurred on D satellite (SAT) chromosomes bearing a C genome translocation.
Leggett
J M
JM
Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, UK. mike.leggett@bbsrc.ac.uk
Perret
S J
SJ
Harper
J
J
Morris
P
P
D018554:Avena / Q000235:genetics*; D015139:Blotting, Southern; D002874:Chromosome Mapping / Q000379:methods*; D018628:Gene Dosage; D017343:Genes, Plant; D006579:Heterozygote; D017404:In Situ Hybridization, Fluorescence; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D011123:Polyploidy; D014170:Transformation, Genetic; D019076:Transgenes