Integration and inheritance stability of foreignBt toxin gene in the bivalent insect-resistant transgenic cotton plants

Abstract

Genetic and expressional stability ofBt toxin gene is crucial for the breeding of insect-resistant transgenic cotton varieties and their commercialization. Genomic Southern blot analysis of R3, R4 and R5 generations of bivalent transgenic insect-resistant cotton plants was done in order to determine the integration, the copy number and the inheritance stability ofBt toxin gene in the transgenic cotton plants. The results indicated that there was a 4.7 kb positive band in the Southern blot when the genomic DNA of the bivalent transgenic insect-resistant cotton plants and the positive control (the plasmid) were digested withHind III respectively. This result proved that theBt toxin gene had been integrated into the genome of the cotton in full length. There is only oneXho I restriction site in theBt toxin gene. Southern blot analysis indicated that many copies ofBt toxin gene had been integrated into the genome of the cotton when the genomic DNA of transgenic plants was digested withXho I. Among them, there were four copies (about 17.7, 8, 5.5 and 4.7 kb in size) existing in all the tested plants of R3, R4 and R5 generations. The preliminary conclusion was that there were more than four copies ofBt toxin gene integrated into the genome of the cotton, among them, more than one copy can express and inherit steadily. This result provides a scientific basis for the breeding of the bivalent insect-resistant transgenic cotton plants and its commercialization.