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.
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 degrees 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.
Institute of Experimental Botany, Academy of Sciences of Czech Republic, Na Karlovce 1a, 160 00 Prague 6, Czech Republic. gichner@ueb.cas.cz
Tomás
T
Gichner
Anita
A
Mukherjee
Jirí
J
Velemínský
D000477:Alkylating Agents / Q000633:toxicity*; D001583:Benzoxazoles / Q000737:chemistry; D020552:Comet Assay; D018744:DNA, Plant / Q000032:analysis* / Q000737:chemistry; D003850:Deoxyribonuclease I / Q000378:metabolism / Q000494:pharmacology*; D004996:Ethidium / Q000737:chemistry; D005020:Ethyl Methanesulfonate / Q000633:toxicity*; D005456:Fluorescent Dyes; D006358:Hot Temperature; D007211:Indoles / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry / Q000187:drug effects; D018517:Plant Roots / Q000737:chemistry / Q000187:drug effects; D011806:Quinolinium Compounds / Q000737:chemistry; D036226:Seedlings / Q000737:chemistry / Q000187:drug effects; D059010:Single-Cell Analysis; D013194:Staining and Labeling / Q000379:methods*; D014026:Nicotiana / Q000737:chemistry / Q000187:drug effects*
D000477:Alkylating Agents; D001583:Benzoxazoles; D018744:DNA, Plant; D005456:Fluorescent Dyes; D007211:Indoles; D011806:Quinolinium Compounds; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; C007293:DAPI; D005020:Ethyl Methanesulfonate; D003850:Deoxyribonuclease I; D004996:Ethidium