Risk assessment of cadmium-contaminated soil on plant DNA damage using RAPD and physiological indices
Risk assessment of cadmium-contaminated soil on plant DNA damage using RAPD and physiological indices
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利用RAPD和生理指标评估镉污染土壤对植物DNA损伤的风险
DOI：
10.1016/j.jhazmat.2008.04.038
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发表时间：
2009-01-30
影响因子：
13.6
通讯作者：
Han, Y. P.
中科院分区：
文献类型：
--
作者：
Liu, Wan;Yang, Y. S.;Han, Y. P.
土壤污染物影响评价是环境质量研究和污染土地修复的重要内容。建立了一种随机扩增多态性DNA （RAPD）指纹图谱技术，用于检测重金属污染土壤中基因毒素诱导的植物DNA损伤。本研究比较了土壤中镉（Cd）污染对大麦幼苗在分子水平和群体水平上的影响。结果表明，随着Cd浓度的升高，大麦幼苗根系生长下降，根尖总可溶性蛋白水平升高。在RAPD分析中，9个GC含量为60-70%的10碱基对随机RAPD引物（十聚体）产生了独特的多态性带模式，随后在对照幼苗的根尖上产生了129个分子量为144-2639碱基对的RAPD片段。9个引物的结果表明，与对照苗相比，Cd处理后根尖RAPD谱的变化包括条带强度的变化以及条带的增加或减少。在10、20和40 mg L-1 Cd的9个引物（1 - 4个新的聚合酶链反应产物）中，几乎出现了分子量约为154 ~ 2245 bp的新扩增片段，并且随着Cd浓度的增加，9个引物缺失的条带数量增加。这些结果表明，在上述镉浓度下，反映RAPD谱变化的基因组模板稳定性受到了显著影响，其结果优于生长和可溶性蛋白水平等传统指标。RAPD检测到的DNA多态性可作为检测土壤镉胁迫对植物遗传毒性的一种合适的生物标志物。RAPD分析作为一种风险评估工具，可用于土壤镉危害的表征。(C) 2008 Elsevier B.V.版权所有
Impact assessment of contaminants in soil is an important issue in environmental quality study and remediation of contaminated land. A random amplified polymorphic DNA (RAPD) 'fingerprinting' technique was exhibited to detect genotoxin-induced DNA damage of plants from heavy metal contaminated soil. This study compared the effects occurring at molecular and population levels in barley seedlings exposed to cadmium (Cd) contamination in soil. Results indicate that reduction of root growth and increase of total soluble protein level in the root tips of barley seedlings occurred with the ascending Cd concentrations. For the RAPD analyses, nine 10-base pair (bp) random RAPD primers (decamers) with 60-70% GC content were found to produce unique polymorphic band patterns and subsequently were used to produce a total of 129 RAPD fragments of 144-2639 base pair in molecular size in the root tips of control seedlings. Results produced from nine primers indicate that the changes occurring in RAPD profiles of the root tips following Cd treatment included alterations in band intensity as well as gain or loss of bands compared with the control seedlings. New amplified fragments at molecular size from approximately 154 to 2245 bp appeared almost for 10, 20 and 40 mg L-1 Cd with 9 primers (one-four new polymerase chain reaction, (PCR) products), and the number of missing bands enhanced with the increasing Cd concentration for nine primers. These results suggest that genomic template stability reflecting changes in RAPD profiles were significantly affected and it compared favourably with the traditional indices such as growth and soluble protein level at the above Cd concentrations. The DNA polymorphisms detected by RAPD can be applied as a suitable biomarker assay for detection of the genotoxic effects of Cd stress in soil on plants. As a tool in risk assessment the RAPD assay can be used in characterisation of Cd hazard in soil. (C) 2008 Elsevier B.V. All rights reserved.