A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants.
Transposition of the Anthirrinum majus Tam3 element and the Zea mays Ac element has been monitored in petunia and tobacco plants. Plant vectors were constructed with the transposable elements cloned into the leader sequence of a marker gene. Agrobacterium tumefaciens-mediated leaf disc transformation was used to introduce the transposable element constructs into plant cells. In transgenic plants, excision of the transposable element restores gene expression and results in a clearly distinguishable phenotype. Based on restored expression of the hygromycin phosphotransferase II (HPTII) gene, we established that Tam3 excises in 30% of the transformed petunia plants and in 60% of the transformed tobacco plants. Ac excises from the HPTII gene with comparable frequencies (30%) in both plant species. When the beta-glucuronidase (GUS) gene was used to detect transposition of Tam3, a significantly lower excision frequency (13%) was found in both plant species. It could be shown that deletion of parts of the transposable elements Tam3 and Ac, removing either one of the terminal inverted repeats (TIR) or part of the presumptive transposase coding region, abolished the excision from the marker genes. This demonstrates that excision of the transposable element Tam3 in heterologous plant species, as documented for the autonomous element Ac, also depends on both properties. Southern blot hybridization shows the expected excision pattern and the reintegration of Tam3 and Ac elements into the genome of tobacco plants.
Haring
M A
MA
Free University, Dept. of Genetics, Amsterdam, Netherlands.
Gao
J
J
Volbeda
T
T
Rommens
C M
CM
Nijkamp
H J
HJ
Hille
J
J
D016960:Agrobacterium tumefaciens / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004251:DNA Transposable Elements; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics*; D010947:Plants, Toxic; D015183:Restriction Mapping; D014026:Nicotiana / Q000235:genetics; D003313:Zea mays / Q000235:genetics
D004251:DNA Transposable Elements; D004247:DNA; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D005966:Glucuronidase