Application of random amplified polymorphic DNA (RAPD) to detect the genotoxic effect of heavy metals

Abstract

This paper presents the results of a study on the influence of lead, copper, manganese and cadmium on DNA integrity in plant cells. Plants, as biological indicators, can measure the potential effects of pollutants when they are used to measure effects of heavy metals. The genotoxicity of heavy metals in kidney-bean (Phaseolus vulgaris) seedlings was subjected to RAPD (random amplified polymorphic DNA) analysis. An RAPD 'fingerprinting' technique was used to detect DNA damage in the kidney-bean seedlings treated with two selected heavy metals at concentrations of 150 and 350 mg x l(-1). Polymorphisms became evident as the presence and/or absence of DNA fragments in treated samples compared with the untreated one. At 350 mg x l(-1), a high number of both missing bands and new amplified fragment were observed. Results suggested that a qualitative measure reflecting changes in RAPD profiles were significantly affected at higher concentrations (350 mg x l(-1)) of the tested heavy metals. A total of 467 RAPD fragments in RAPD profiles were detected by using six random primers (decamers) and 224 of these fragments showed polymorphism. There was a distinct distance between the band patterns of treated plants and the control samples when the cluster method was applied. In addition, the result derived from numerical analysis revealed a considerable distance between the band pattern of the plant samples treated with 350 mg x l(-1) heavy metals and the control sample. Finally, a comparison between untreated and treated genomes shows that RAPD analysis can be used to evaluate how the environmental pollutants modify the structure of DNA in living organisms.

Mineral nutrient imbalance, total antioxidants level and DNA damage in common bean (Phaseolus vulgaris L.) exposed to heavy metals

“…This variable correlations can be explained by a range of influencing factors, including the diversity among samples, variable reaction kinetics, presence and activity of enzymes responsible for mediating oxidative stress, (Jaio and Wang 2000;Wu et al 2004;Ozgen et al 2006) and etc. In the samples treated with Ni and Cd as non-redox metals, correspondingly to previously published facts we detected high levels of (Enan 2006;Kekec et al 2010;Cenkci et al 2009). The RAPD, PCR based assay described here is fast, reliable, and easy to conduct in any laboratory for assessment of environmental hazardous metals on plants.…”

“…This plant is exposed to heavy metals in its natural environment as a result of various human activities. Bean is a diploid (2n=22) and it has been widely used in physiological and molecular analysis in toxicology (Enan 2006;Kekec et al 2010). Seeds were surface-sterilized with 75 % (v/v) ethanol for 5 min, followed by 10 % (m/v) sodium hypochlorite for 10 min.…”

“…Advantages of measuring effects of genotoxic chemicals directly on DNA are mainly related to the sensitivity and short response time. DNA based techniques like Random Amplified Polymorphic DNA (RAPD) is used to evaluate the variation at the DNA level and can clearly be shown when comparing DNA fingerprints from untreated and treated individuals to genotoxic agents (Savva 1998;Atienzar and Jha 2006;Enan 2006;Kekec et al 2010).…”

QUALITATIVE ASSESSMENT OF CADMIUM STRESS USING GENOME TEMPLATE STABILITY IN Hordeum vulgare

“…RAPD analysis have been reported as more sensitive test than classic genotoxic tests since it was capable of detecting temporary DNA changes that may not finally manifest themselves as mutations (Labra et al, 2003;Liu et al, 2007). The present results of Cd exposure in barley seedlings are consistent with the earlier studies on other organisms (Enan, 2006;Liu et al, 2007;Cambier et al, 2010).…”

Alterations in Biomarkers Associated with Sterility in Pectinophora gossypiella (Saunders) Induced by Gamma Irradiation

“…Recently, several selective and sensitive assays developed, such as restriction fragment length polymorphism, quantitative traits loci, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphisms, simple sequence repeat, variable number of tandem repeats for DNA analysis in ecogenotoxicology as a result of the advances in molecular biology 9 . RAPD-PCR is one of the most feasible methods or techniques used for detection of DNA damage and mutations comparing DNA fingerprints from untreated and treated samples 10,11 . Previous studies have shown the biochemical and biological effect of gamma radiation doses (100 and 150 Gray) on F1 adult male and female almond moths, Ephestia cautella (Walker) (Pyralidae: Lepidoptera) descendant of irradiated parental male pupae 12 , and the variation induced in elemental contents in the body tissues of Galleria mellonella adult males and females parents irradiated with 50 and 150 Gy and their F1 generation 13 .…”