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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Guo, S. D., Ni, W. C. et al., The application of the fused gene encoding the insecticidal protein and the expression vector (in Chinese), 1995, 12.28 Patent ZL 95119563.8,C12N.
Guo, S. D., Cui, H. Z., Xia, L. Q. et al., The study of the bivalent insect-resistant transgenic cotton plants, Scientia Agriculturia Sinica (in Chinese), 1999, 32(3): 1.
Tang, C. M., Sun, J., Zhu, X. F. et al., The genetic analysis of the insect-resistant activity of the threeBt transgenic cotton plants in China, Chinese Science Bulletin, 1999, 44 (19): 2064.
Zhang, T, Z., Tang, C. M., The utilization and dissemination of theBt transgenic cotton plants and the management of the insect-resistance activity, Chinese Science Bulletin, 2000, 45(2): 119.
Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, 1989.
Peterson, A. H., A rapid method for the extraction of cotton genomic DNA suitable for RFLP or PCR analysis, Plant Molecular Biology Reporter, 1993, 11: 122.
Peng, J., Wen, F., Lister, R. L. et al., Inheritance of gusA and neo genes in transgenic rice, Plant Mol. Biol., 1995, 27: 91.
Rathore, K. S., Chowdhury, V. K., Hodges, T. K., Use of bar as a selectable marker gene and for the production of herbicide resistant rice plants from protoplast, Plant Mol. Biol., 1993, 21: 871.
Cao, J., Duan, X., Mcelroy, D. et al., Regeneration of herbicide resistant transgenic rice plants following miroprojectile mediated transformation of suspension culture cells, Plant Cell. Rep., 1992, 11: 586.
Matassi, G., Montero, L. M., Salinas, J. et al., The isochore organization and the compositional distribution of homologous coding sequences in the nuclear genome of plants, Nucleic Acids Research, 1989, 17: 5273.
Guo, W. Z., Zhang, T. Z., Zhu, X. F. et al., Variation induced by DNA rearrangement in a transgenicBt+CpTI cotton strain, Chinese Science Bulletin, 2001, 46(7): 559.
Finnegan, J., McElroy, D., Transgene inactivation: plants fight bank! Bio/Technology, 1994, 12: 883.
Zhou, G. Y., Wang, J., Gong, Z. et al., Molecular breeding of agriculture: a technique for introducing exogenouse DNA into plants after self-pollination, Scientia Agriculturia Sinica, 1988, 21: 1.
Ni, W. C., Zhang, Z. L., Guo, S. D., The cultivation of the insect-resistant transgenic cotton plants, Scientia Agriculturia Sinica, 31(2): 8.
Langridge, P., Brettschneider, R., Lazzeri, P. et al., Transformation of cereals via Agrobacterium and the pollen pathway: a critical assessment, The Plant Journal, 1992, 2(4): 631.
Flavell, R. B., Inativation of gene expression in plants as consequence of specific sequence duplication, Proc. Natl. Acad., Sci. USA, 1994, 91: 3490.
Hobbs, S. L. A., Thomas, D. W., Delong, C. M. O., Transgene copy number can be positively or negatively associated with transgene expression, Plant Mol. Bio., 1993, 21: 17.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Xia, L., Guo, S. Integration and inheritance stability of foreignBt toxin gene in the bivalent insect-resistant transgenic cotton plants. Chin.Sci.Bull. 46, 1372–1375 (2001). https://doi.org/10.1007/BF03183392
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF03183392