Abstract
We show that major chromosomal rearrangements can occur upon T-DNA transformation of Arabidopsis thaliana. In the ACL4 line, two T-DNA insertion loci were found; one is a tandem T-DNA insert in a head-to-head orientation, and the other is a truncated insert with only the left part of the T-region. The four flanking DNA regions were isolated and located on the Arabidopsis chromosomes; for both inserts, one side of the T-DNA maps to chromosome 2, whereas the other side maps to chromosome 3. Both chromosome 3 flanking regions map to the same location, despite a 1.4-kb deletion at this point, whereas chromosome 2 flanking regions are located 40 cM apart on the bottom arm of chromosome 2. These results strongly suggest a reciprocal translocation between chromosomes 2 and 3, with the breakpoints located at the T-DNA insertion sites. The interchanged fragments roughly correspond to the 20-cM distal ends of both chromosomes. Moreover, a large inversion, spanning 40 cM on the genetic map, occurs on the bottom arm of chromosome 2. This was confirmed by genetic analyses that demonstrated a strong reduction of recombination in the inverted region. Models for T-DNA integration and the consequences for T-DNA tagging are discussed in light of these results.
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- Alexander M. P. Differential staining of aborted and nonaborted pollen. Stain Technol. 1969 May;44(3):117–122. doi: 10.3109/10520296909063335. [DOI] [PubMed] [Google Scholar]
- Azpiroz-Leehan R., Feldmann K. A. T-DNA insertion mutagenesis in Arabidopsis: going back and forth. Trends Genet. 1997 Apr;13(4):152–156. doi: 10.1016/s0168-9525(97)01094-9. [DOI] [PubMed] [Google Scholar]
- Castle L. A., Errampalli D., Atherton T. L., Franzmann L. H., Yoon E. S., Meinke D. W. Genetic and molecular characterization of embryonic mutants identified following seed transformation in Arabidopsis. Mol Gen Genet. 1993 Dec;241(5-6):504–514. doi: 10.1007/BF00279892. [DOI] [PubMed] [Google Scholar]
- Creusot F., Fouilloux E., Dron M., Lafleuriel J., Picard G., Billault A., Le Paslier D., Cohen D., Chabouté M. E., Durr A. The CIC library: a large insert YAC library for genome mapping in Arabidopsis thaliana. Plant J. 1995 Nov;8(5):763–770. doi: 10.1046/j.1365-313x.1995.08050763.x. [DOI] [PubMed] [Google Scholar]
- De Neve M., De Buck S., Jacobs A., Van Montagu M., Depicker A. T-DNA integration patterns in co-transformed plant cells suggest that T-DNA repeats originate from co-integration of separate T-DNAs. Plant J. 1997 Jan;11(1):15–29. doi: 10.1046/j.1365-313x.1997.11010015.x. [DOI] [PubMed] [Google Scholar]
- Deng X. W., Caspar T., Quail P. H. cop1: a regulatory locus involved in light-controlled development and gene expression in Arabidopsis. Genes Dev. 1991 Jul;5(7):1172–1182. doi: 10.1101/gad.5.7.1172. [DOI] [PubMed] [Google Scholar]
- Edwards K., Johnstone C., Thompson C. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 1991 Mar 25;19(6):1349–1349. doi: 10.1093/nar/19.6.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldmann K. A., Coury D. A., Christianson M. L. Exceptional segregation of a selectable marker (KanR) in Arabidopsis identifies genes important for gametophytic growth and development. Genetics. 1997 Nov;147(3):1411–1422. doi: 10.1093/genetics/147.3.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gheysen G., Villarroel R., Van Montagu M. Illegitimate recombination in plants: a model for T-DNA integration. Genes Dev. 1991 Feb;5(2):287–297. doi: 10.1101/gad.5.2.287. [DOI] [PubMed] [Google Scholar]
- Mayerhofer R., Koncz-Kalman Z., Nawrath C., Bakkeren G., Crameri A., Angelis K., Redei G. P., Schell J., Hohn B., Koncz C. T-DNA integration: a mode of illegitimate recombination in plants. EMBO J. 1991 Mar;10(3):697–704. doi: 10.1002/j.1460-2075.1991.tb07999.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Negruk V., Eisner G., Lemieux B. Addition-deletion mutations in transgenic Arabidopsis thaliana generated by the seed co-cultivation method. Genome. 1996 Dec;39(6):1117–1122. doi: 10.1139/g96-140. [DOI] [PubMed] [Google Scholar]
- Ohba T., Yoshioka Y., Machida C., Machida Y. DNA rearrangement associated with the integration of T-DNA in tobacco: an example for multiple duplications of DNA around the integration target. Plant J. 1995 Jan;7(1):157–164. doi: 10.1046/j.1365-313x.1995.07010157.x. [DOI] [PubMed] [Google Scholar]
- Park S. H., Pinson S. R., Smith R. H. T-DNA integration into genomic DNA of rice following Agrobacterium inoculation of isolated shoot apices. Plant Mol Biol. 1996 Dec;32(6):1135–1148. doi: 10.1007/BF00041397. [DOI] [PubMed] [Google Scholar]
- Ray S. M., Park S. S., Ray A. Pollen tube guidance by the female gametophyte. Development. 1997 Jun;124(12):2489–2498. doi: 10.1242/dev.124.12.2489. [DOI] [PubMed] [Google Scholar]
- Round E. K., Flowers S. K., Richards E. J. Arabidopsis thaliana centromere regions: genetic map positions and repetitive DNA structure. Genome Res. 1997 Nov;7(11):1045–1053. doi: 10.1101/gr.7.11.1045. [DOI] [PubMed] [Google Scholar]
- Takano M., Egawa H., Ikeda J. E., Wakasa K. The structures of integration sites in transgenic rice. Plant J. 1997 Mar;11(3):353–361. doi: 10.1046/j.1365-313x.1997.11030353.x. [DOI] [PubMed] [Google Scholar]
- Zachgo E. A., Wang M. L., Dewdney J., Bouchez D., Camilleri C., Belmonte S., Huang L., Dolan M., Goodman H. M. A physical map of chromosome 2 of Arabidopsis thaliana. Genome Res. 1996 Jan;6(1):19–25. doi: 10.1101/gr.6.1.19. [DOI] [PubMed] [Google Scholar]
- Zambryski P., Tempe J., Schell J. Transfer and function of T-DNA genes from agrobacterium Ti and Ri plasmids in plants. Cell. 1989 Jan 27;56(2):193–201. doi: 10.1016/0092-8674(89)90892-1. [DOI] [PubMed] [Google Scholar]