Menke M, Chen I, Angelis K J, Schubert I
Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany.
Mutat Res. 2001 Jun 27;493(1-2):87-93. doi: 10.1016/s1383-5718(01)00165-6.
The three protocols of the comet assay A/N, A/A and N/N were for the first time applied to the plant species Arabidopsis thaliana. The purpose of the experiments was to establish conditions for genotoxic exposure causing DNA damage in Arabidopsis nuclei. This is required for comprehensive gene expression profiling with the intention to screen for genes involved in response of Arabidopsis cells to genotoxic stress. Five chemicals belonging to different classes of mutagens (the monofunctional alkylating agents N-methyl-N-nitrosourea and methyl methanesulfonate, the polyfunctional alkylating agent mitomycin C, the radiomimetic bleomycin and the herbicide maleic hydrazide) were tested. Except for maleic hydrazide, dose-dependent increases in DNA damage were found using the A/N comet assay protocol. While a rapid repair of bleomycin-mediated SSBs and DSBs was found, no significant reduction of DNA migration was observed up to 48h after treatment with the monofunctional alkylating agents.
彗星试验的三种方案A/N、A/A和N/N首次应用于植物拟南芥。实验目的是建立导致拟南芥细胞核DNA损伤的遗传毒性暴露条件。这对于全面的基因表达谱分析是必要的，目的是筛选参与拟南芥细胞对遗传毒性应激反应的基因。测试了五种属于不同诱变剂类别的化学物质（单功能烷基化剂N-甲基-N-亚硝基脲和甲磺酸甲酯、多功能烷基化剂丝裂霉素C、放射模拟物博来霉素和除草剂马来酰肼）。除马来酰肼外，使用A/N彗星试验方案发现DNA损伤呈剂量依赖性增加。虽然发现博来霉素介导的单链断裂和双链断裂能快速修复，但在用单功能烷基化剂处理后48小时内未观察到DNA迁移的显著减少。
