Erturk Filiz Aygun, Ay Hilal, Nardemir Gokce, Agar Guleray
Department of Biology, Atatürk University, Erzurum, Turkey.
Toxicol Ind Health. 2013 Aug;29(7):662-71. doi: 10.1177/0748233712442709. Epub 2012 Apr 12.
Assessment of DNA damages stemming from toxic chemicals is an important issue in terms of genotoxicology. In this study, maize (Zea mays L.) seedlings were used for screening the genotoxic effects of cobalt (Co) and nickel (Ni) treatments at various concentrations (5 mM, 10 mM, 20 mM and 40 mM). For this purpose, randomly amplified polymorphic DNA (RAPD) technique was applied to genomic DNA extracted from metal-exposed and unexposed plant materials. Besides, changes in total protein contents were screened by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. For RAPD analysis, 16 RAPD primers were found to produce unique polymorphic band profiles on different concentrations of Co-/Ni-treated maize seedlings. Increased polymorphism resulting from the appearance of new bands or disappearance of normal bands was observed with increasing concentration of Co and Ni treatments. Genomic template stability, a qualitative measurement of changes in RAPD patterns of genomic DNA, decreased with increasing metal concentration. In SDS-PAGE analysis, it was observed that the total soluble protein content decreased by Co treatment, while it increased by Ni treatment. The results obtained from this study revealed that RAPD profiles and total soluble protein levels can be applied to detect genotoxicity, and these analyses can offer useful biomarker assays for the evaluation of genotoxic effects on Co- and Ni-polluted plants.
从遗传毒理学角度来看，评估有毒化学物质造成的DNA损伤是一个重要问题。在本研究中，使用玉米（Zea mays L.）幼苗来筛选不同浓度（5 mM、10 mM、20 mM和40 mM）的钴（Co）和镍（Ni）处理的遗传毒性效应。为此，将随机扩增多态性DNA（RAPD）技术应用于从暴露于金属和未暴露于金属的植物材料中提取的基因组DNA。此外，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析筛选总蛋白含量的变化。对于RAPD分析，发现16条RAPD引物在不同浓度的Co/Ni处理的玉米幼苗上产生独特的多态性条带图谱。随着Co和Ni处理浓度的增加，观察到由于新条带的出现或正常条带的消失而导致的多态性增加。基因组模板稳定性是对基因组DNA的RAPD模式变化的定性测量，随着金属浓度的增加而降低。在SDS-PAGE分析中，观察到Co处理使总可溶性蛋白含量降低，而Ni处理使其增加。本研究获得的结果表明，RAPD图谱和总可溶性蛋白水平可用于检测遗传毒性，并且这些分析可为评估Co和Ni污染植物的遗传毒性效应提供有用的生物标志物检测方法。
