Comparison of DNA damage in plants as measured by single cell gel electrophoresis and somatic leaf mutations induced by monofunctional alkylating agents.
The use of single cell gel electrophoresis (SCGE) has recently been applied to plant systems. We optimized the experimental conditions for SCGE analysis using nuclei isolated from different tissues of intact plants. Concentration-response curves of genomic DNA migration were analyzed in intact plants treated with the monofunctional alkylating agents ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), N-ethyl-N-nitrosourea (ENU), and N-methyl-N-nitrosourea (MNU). These data were used to calibrate SCGE tail moment values to induced somatic mutation in plant leaves. We used a genotoxicity index to compare genomic DNA damage and the induction of somatic mutation in the leaf tissues. The rank order of the genotoxic potency of these alkylating agents assayed by SCGE was MNU >> MMS > ENU > EMS. The rank order for the mutagenic potency of these agents was MNU >> ENU congruent with MMS > EMS. The data demonstrate the utility of SCGE analysis in plant systems. The use of SCGE will permit a larger range of plants for use as in situ environmental monitors. Also, this approach may be used to search for crop plant germplasm accessions with enhanced genomic stability. We investigated whether the intragenomic distributions of DNA damage induced by these alkylating agents were uniform and random. When a plot of the ratio of the %tail DNA and tail length versus the concentration of the test mutagen was generated, the induced SCGE data deviated from a random distribution of genomic DNA damage.
Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
T
T
Gichner
O
O
Ptácek
D A
DA
Stavreva
M J
MJ
Plewa
D000477:Alkylating Agents / Q000633:toxicity*; D004249:DNA Damage; D018744:DNA, Plant / Q000737:chemistry / Q000187:drug effects; D004586:Electrophoresis / Q000379:methods*; D004784:Environmental Monitoring; D005020:Ethyl Methanesulfonate / Q000633:toxicity; D005038:Ethylnitrosourea / Q000633:toxicity; D008741:Methyl Methanesulfonate / Q000633:toxicity; D008770:Methylnitrosourea / Q000633:toxicity; D009153:Mutagens / Q000633:toxicity*; D009690:Nucleic Acid Conformation; D018515:Plant Leaves / Q000187:drug effects* / Q000235:genetics; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics; D010947:Plants, Toxic; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics
D000477:Alkylating Agents; D018744:DNA, Plant; D009153:Mutagens; D008770:Methylnitrosourea; D005020:Ethyl Methanesulfonate; D008741:Methyl Methanesulfonate; D005038:Ethylnitrosourea