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Application of the comet assay to measure DNA damage induced by UV radiation in the hydrophyte, Spirodela polyrhiza
Author(s) -
Jiang Lei,
Wang Yan,
Li Shaoshan
Publication year - 2007
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.2006.00820.x
Subject(s) - comet assay , dna damage , dna , endonuclease , gel electrophoresis , pyrimidine dimer , biology , plant cell , microbiology and biotechnology , hydrogen peroxide , biophysics , chemistry , biochemistry , gene
The single‐cell gel electrophoresis or comet assay is now widely used to detect DNA damage in animal cells induced by radiation or chemicals. Here, we apply the comet assay to measure ultraviolet (UV)‐B‐induced DNA damage in plant cells. The accepted animal cell protocol for the comet assay was modified to adapt it to plant cells. The major modifications were conversion of the plant cells to protoplasts and the use of T4 endonuclease V. As a positive control hydrogen peroxide was applied. Significant DNA damage was detected at 100 μ M H 2 O 2 . This type of DNA damage was not affected by T4 endonuclease V treatment, which implies that the mechanism of H 2 O 2 ‐induced DNA damage was different from UV‐B‐induced DNA damage. Our results also indicate that both UV‐A and UV‐B radiation can induce DNA single‐strand breaks in plant cells, while UV‐B was more effective than UV‐A for inducing pyrimidine dimer formation.