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. 1992 Mar;4(3):319–332. doi: 10.1105/tpc.4.3.319

Somatic and Meiotic Chromosomal Recombination between Inverted Duplications in Transgenic Tobacco Plants.

J Tovar 1, C Lichtenstein 1
PMCID: PMC160132  PMID: 12297648

Abstract

Homologous recombination has been extensively studied in bacteria, yeast, and more recently in animal cells, but little is known about this process in plants. We present here an analysis of meiotic and somatic chromosomal recombination between closely linked inverted duplications located on a single chromosomal region in tobacco. Transgenic tobacco lines were constructed by Agrobacterium transformation with plasmid vectors containing a functional hygromycin phosphotransferase (hyg) selectable marker flanked by a pair of defective neomycin phosphotransferase (neo) genes positioned as inverted repeats. As each neo gene is mutated in a different site, recombination between the two defective genes can be detected following selection for kanamycin-resistant plant cells. The recombination substrates were designed to allow investigation into the nature of molecular events underlying homologous recombination by restriction endonuclease analysis. Chromosomal recombination was studied in mitotically dividing cells (cultured leaf mesophyll cells) and after meiosis (germinated seedlings). Spontaneous somatic recombinants were recovered at frequencies between ~3 x 10-5 to 10-6 events per cell. Low dose [gamma] irradiation of somatic cells resulted in a threefold maximum increase in the recovery of recombinants. Recombinants were also detected at low frequency when transgenic T3 seeds were germinated under kanamycin selection. DNA gel blot analyses demonstrated that homologous recombination occurred mainly as gene conversion unassociated with reciprocal exchange, although a variety of other events including gene coconversion were also observed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. An G. Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol. 1986 May;81(1):86–91. doi: 10.1104/pp.81.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baur M., Potrykus I., Paszkowski J. Intermolecular homologous recombination in plants. Mol Cell Biol. 1990 Feb;10(2):492–500. doi: 10.1128/mcb.10.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dooner H. K., Kermicle J. L. The Transposable Element Ds Affects the Pattern of Intragenic Recombination at the bz and R Loci in Maize. Genetics. 1986 May;113(1):135–143. doi: 10.1093/genetics/113.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fu T. K., Sears E. R. The Relationship between Chiasmata and Crossing over in TRITICUM AESTIVUM. Genetics. 1973 Oct;75(2):231–246. doi: 10.1093/genetics/75.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gal S., Pisan B., Hohn T., Grimsley N., Hohn B. Genomic homologous recombination in planta. EMBO J. 1991 Jun;10(6):1571–1578. doi: 10.1002/j.1460-2075.1991.tb07677.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Game J. C., Zamb T. J., Braun R. J., Resnick M., Roth R. M. The Role of Radiation (rad) Genes in Meiotic Recombination in Yeast. Genetics. 1980 Jan;94(1):51–68. doi: 10.1093/genetics/94.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HOLLIDAY R. THE INDUCTION OF MITOTIC RECOMBINATION BY MITOMYCIN C IN USTILAGO AND SACCHAROMYCES. Genetics. 1964 Sep;50:323–335. doi: 10.1093/genetics/50.3.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Katz E. R., Kao V. Evidence for mitotic recombination in the cellular slime mold Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4025–4026. doi: 10.1073/pnas.71.10.4025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Letsou A., Liskay R. M. Effect of the molecular nature of mutation on the efficiency of intrachromosomal gene conversion in mouse cells. Genetics. 1987 Dec;117(4):759–769. doi: 10.1093/genetics/117.4.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liskay R. M., Stachelek J. L., Letsou A. Homologous recombination between repeated chromosomal sequences in mouse cells. Cold Spring Harb Symp Quant Biol. 1984;49:183–189. doi: 10.1101/sqb.1984.049.01.021. [DOI] [PubMed] [Google Scholar]
  12. MURRAY N. E. Polarized recombination and fine structure within the me-2 gene of Neurospora crassa. Genetics. 1963 Sep;48:1163–1183. doi: 10.1093/genetics/48.9.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McClintock B. The Stability of Broken Ends of Chromosomes in Zea Mays. Genetics. 1941 Mar;26(2):234–282. doi: 10.1093/genetics/26.2.234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nelson O. E. The WAXY Locus in Maize. II. the Location of the Controlling Element Alleles. Genetics. 1968 Nov;60(3):507–524. doi: 10.1093/genetics/60.3.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Neuffer M. G. Crossing over in heterozygotes carrying different mutable alleles at the A1 locus in maize. Genetics. 1965 Sep;52(3):521–528. doi: 10.1093/genetics/52.3.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Peterhans A., Schlüpmann H., Basse C., Paszkowski J. Intrachromosomal recombination in plants. EMBO J. 1990 Nov;9(11):3437–3445. doi: 10.1002/j.1460-2075.1990.tb07551.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith A. J., Berg P. Homologous recombination between defective neo genes in mouse 3T6 cells. Cold Spring Harb Symp Quant Biol. 1984;49:171–181. doi: 10.1101/sqb.1984.049.01.020. [DOI] [PubMed] [Google Scholar]
  20. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]

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