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THE PLANT CELL, Vol 6, Issue 11 1635-1641, Copyright © 1994 by American Society of Plant Biologists
DNA Damage Levels Determine Cyclobutyl Pyrimidine Dimer Repair Mechanisms in Alfalfa Seedlings
F. E. Quaite, S. Takayanagi, J. Ruffini, J. C. Sutherland and B. M. Sutherland
Biology Department, Brookhaven National Laboratory, Upton, New York 11973-5000
Ultraviolet radiation in sunlight damages DNA in plants, but little is
understood about the types, lesion capacity, and coordination of repair
pathways. We challenged intact alfalfa seedlings with UV doses that induced
different initial levels of cyclobutyl pyrimidine dimers and measured
repair by excision and photoreactivation. By using alkaline gel
electrophoresis of nonradioactive DNAs treated with a cyclobutyl pyrimidine
dimer-specific UV endonuclease, we quantitated ethidium-stained DNA by
electronic imaging and calculated lesion frequencies from the number
average molecular lengths. At low initial dimer frequencies (less than ~30
dimers per million bases), the seedlings used only photoreactivation to
repair dimers; excision repair was not significant. At higher damage
levels, both excision and photorepair contributed significantly. This
strategy would allow plants with low damage levels to use error-free repair
requiring only an external light energy source, whereas seedlings subjected
to higher damage frequencies could call on additional repair processes
requiring cellular energy. Characterization of repair in plants thus
requires an investigation of a range of conditions, including the level of
initial damage.
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