DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase-I

• Published in: Mutation research Vol. 605; no. 1; pp. 17 - 21
• Main Authors: Gichner, Tomáš, Mukherjee, Anita, Velemínský, Jiří
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 16.06.2006; Elsevier
• Subjects: Alkylating Agents - toxicity; Benzoxazoles - chemistry; Biological and medical sciences; Comet Assay; Deoxyribonuclease I - metabolism; Deoxyribonuclease I - pharmacology; DNA migration; DNA, Plant - analysis; DNA, Plant - chemistry; Ethidium - chemistry; Ethyl Methanesulfonate - toxicity; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; Hot Temperature; Indoles - chemistry; Medical sciences; Nicotiana - chemistry; Nicotiana - drug effects; Nicotiana tabacum; Plant Leaves - chemistry; Plant Leaves - drug effects; Plant Roots - chemistry; Plant Roots - drug effects; Quinolinium Compounds - chemistry; Seedlings - chemistry; Seedlings - drug effects; Single-Cell Analysis; Single-cell gel electrophoresis; Staining and Labeling - methods; Toxicology; Single-cell gel electrophoresis; YOYO-1; DNA migration; DAPI; EMS; Nicotiana tabacum; EtBr; Enzyme; Mutagenicity testing; Esterases; Comet assay; Tobacco; Toxicology; Deoxyribonuclease I; DNA; Hydrolases; Genetics; Hyperthermia
• ISSN: 1383-5718; 1879-3592; 1873-135X
• DOI: 10.1016/j.mrgentox.2006.01.005

We applied the alkaline version of the single-cell gel electrophoresis (comet) assay to roots and leaves of tobacco ( Nicotiana tabacum var. xanthi) seedlings or isolated leaf nuclei treated with: (1) the alkylating agent ethyl methanesulphonate, (2) necrotic heat treatments at 50 °C, and (3) DNase-I. All three treatments induced a dose-dependent increase in DNA migration, expressed as percentage of tail DNA. A comparison of the fluorochrome DNA dyes ethidium bromide, DAPI and YOYO-1 demonstrated that for the alkaline version of the comet assay in plants, the commonly used fluorescent dye ethidium bromide can be used with the same efficiency as DAPI or YOYO-1.