23.
Cotyledons from germinating seeds of the soybean cultivar Peking were inoculated with virulent
Agrobacterium tumefaciens strain A281:pZA-7 which carries a wild type Ti plasmid pTiBo542 and a disarmed Ti plasmid (a binary vector)pZA-7 which contains the
glucuronidase (
uidA) and neomycin phosphotransferase (
nptII) genes. Tumors were produced on all inoculated explants and 82% of these tumor lines were cotransformed by the
nptII gene from the binary vector pZA-7 as shown by PCR analysis (18 of 22 lines tested). Eleven of these 18 lines were also resistant to kanamycin. Eleven lines expressed
-glucuronidase activity (GUS), six of which were also kanamycin resistant. Since there is a high rate of coexpression of genes carried by the binary vector, this system provides a simple and rapid method for the expression of genes of interest in transformed soybean tissue which has been used successfully to test constructs designed for soybean transformation.
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