Abstract
Agrobacterium rhizogenes was assessed as a vehicle for transformation ofLotus corniculatus. Plants were co-transformed usingA. rhizogenes strain LBA 9402 harbouring the bacterial plasmid pRi1855 and the binary transformation vector pJit 73. pRi 1855 transfers both TL and TR sequences, while pJit 73 encodes β-glucuronidase (GUS) and also two selectable marker genes giving resistance to the antibiotics kanamycin and hygromycin. Three primary transformants (lines 1,6 and 12) were subjected to detailed morphological and biochemical analysis and lines 6 and 12 were also analysed at the molecular level. Tissues of both lines 6 and 12 were resistant to hygromycin and expressed GUS. Analysis of various tissues of each line showed a significantly lower GUS activity in line 6 than in line 12. Genetical analysis of progeny produced between control plants and lines 6 and 12 indicated that line 6 had one dose of theuid gene while line 12 had two or more independently segregating doses of the gene. Both lines 6 and 12 contained multiple copies of TL-DNA, while only line 6 was TR positive. In the progeny of lines 6 and 12 there was no evidence for linkage of TL-DNA withuid, while in the progeny of line 6, TR-DNA was under-represented. GUS-positive progeny which were free of both TL and TR sequences were identified from both lines.
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Webb, K.J., Robbins, M.P. & Mizen, S. Expression of GUS in primary transformants and segregation patterns of GUS, TL- and TR-DNA in the T1 generation of hairy root transformants ofLotus corniculatus . Transgenic Research 3, 232–240 (1994). https://doi.org/10.1007/BF02336776
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DOI: https://doi.org/10.1007/BF02336776