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利用随机扩增多态性 DNA(RAPD)分析评估菜豆作为模式植物系统中重金属的遗传毒性。

Assessment of the genotoxicity of heavy metals in Phaseolus vulgaris L. as a model plant system by Random Amplified Polymorphic DNA (RAPD) analysis.

机构信息

Faculty of Medical Sciences, Goce Delčev University, Štip, Macedonia.

出版信息

J Environ Sci Health A Tox Hazard Subst Environ Eng. 2012;47(3):366-73. doi: 10.1080/10934529.2012.645784.

Abstract

Impact assessments of environmental pollutants are important in eco-genotoxicology. A random amplified polymorphic DNA (RAPD) technique was used to detect genotoxicity-induced DNA damage in Phaseolus vulgaris L. from heavy metals at two different concentrations. The results from six 10-base pair (bp) random RAPD primers with 60-70% GC content used, showed a total of 295 RAPD fragments of 700-4000 bp in molecular size in the seedlings of untreated and treated samples, of which only 163 fragments were polymorphic. Polymorphisms became evident as the disappearance and/or appearance of DNA fragments in treated samples compared to the control. A dendrogram constructed using the Numerical Taxonomy and Multivariate Analysis System (NTSYSps), showed that the control group merged with groups treated with CuSO(4)·5H(2)O (150 mg L(-1)) and MnSO(4)·H(2)O (150 mg L(-1)) in a separate cluster. These groups were linked with all of the other samples treated with metals at concentrations of 150 mg L(-1) and CuSO(4)·5H(2)O and Cd(NO(3))(2) at concentrations of 350 mg L(-1). Finally, the samples treated with metals at concentrations of 350 mg L(-1) together with NiSO(4) at the concentration of 150 mg L(-1), clustered separately. The DNA polymorphism detected by RAPD analysis offered a useful biomarker assay for the detection of toxic chemicals genotoxicity in plant model systems.

摘要

环境污染物的影响评估在生态遗传毒理学中非常重要。本研究采用随机扩增多态性 DNA(RAPD)技术,检测不同浓度重金属对菜豆的遗传毒性损伤。实验选用 6 个 10 碱基对(bp)随机 RAPD 引物,GC 含量为 60-70%,共扩增出 700-4000bp 大小的 295 个 RAPD 片段,其中只有 163 个片段表现出多态性。与对照相比,处理样品中 DNA 片段的消失和/或出现使多态性变得明显。通过数值分类和多元分析系统(NTSYSps)构建的聚类图显示,对照组与 CuSO4·5H2O(150mg/L)和 MnSO4·H2O(150mg/L)处理组聚为一类,而这两组又与其他所有用 150mg/L 浓度的金属和 350mg/L 浓度的 CuSO4·5H2O 和 Cd(NO3)2处理的样本聚在一起。最后,用 350mg/L 浓度的金属和 150mg/L 浓度的 NiSO4 处理的样本单独聚为一类。RAPD 分析检测到的 DNA 多态性为植物模型系统中检测有毒化学物质遗传毒性提供了一种有用的生物标志物检测方法。

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