基于RAPD技术检测不同烃链长度咪唑基离子液体对日本三角涡虫的遗传毒性

Genotoxicity Evaluation of Imidazolium-based Ionic Liquids with Different Alkyl-chain Lengths on Dugesia japonica via RAPD

RAPD标记通常用于遗传作图、分类和系统发育研究,该技术也可用于检测DNA损伤或突变。本研究利用RAPD技术检测了4种不同烃链长度的咪唑基离子液体对日本三角涡虫(Dugesia japonica)的遗传毒性。在20条随机引物中共筛选出11条用于RAPD扩增,其中对照组共扩增出65条带。与对照组相比,离子液体处理组的RAPD图谱变化表现在条带亮度增强或减弱以及新带的出现和正常带的消失。各处理组多态性带及变异带的总数随离子液体烃链碳原子数增加而增加,而基因组模板稳定性则随离子液体烃链长度增加而降低,这说明咪唑基离子液体对涡虫有遗传毒性,且随着离子液体烃链长度增加其对涡虫的遗传毒性增强。同时研究结果也证明RAPD分析是一种检测环境污染物对动物DNA损伤的灵敏方法。

Though commonly used in genetic mapping, taxonomic and phylogenetic studies, RAPD markers can also be used to detect DNA damage and mutation. In the present study, genotoxicity of imidazolium-based ionic liquids with different alkyl-chain lengths in planarian Dugesia japonica was evaluated using the RAPD assay. Eleven primers were screened out of 20 RAPD random primers to apply to the analysis. A total of 65 bands were observed in the untreated control planarians. Compared with the control, the ILs treated groups displayed different RAPD patterns in the band intensity, disappearance of normal bands and appearance of new bands. The total number of polymorphic bands and varied bands in treated groups increased with the increase of carbon atoms in the alkyl chain of ILs, while the genome template stability （GTS） decreased. This suggested that imidazolium-based ILs had genotoxic effects on planarians and the genotoxicity increased with the lengthening of the alkyl-chain length of the imidazolium-based ILs. Our results also confirmed that RAPD analysis was a highly sensitive method for the detection of DNA damage in animals induced by environmental pollutants such as toxic chemicals.