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THE PLANT CELL, Vol 4, Issue 3 319-332, Copyright © 1992 by American Society of Plant Biologists
Somatic and Meiotic Chromosomal Recombination between Inverted Duplications in Transgenic Tobacco Plants
J. Tovar and C. Lichtenstein
Centre for Biotechnology, Imperial College of Science, Technology, and Medicine, Exhibition Road, London SW7 2AZ, United Kingdom
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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