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切尔诺贝利大豆幼苗中的DNA损伤、修复监测及表观遗传DNA甲基化变化

DNA damage, repair monitoring and epigenetic DNA methylation changes in seedlings of Chernobyl soybeans.

作者信息

Georgieva Mariyana, Rashydov Namik M, Hajduch Martin

机构信息

Department of Reproduction and Developmental Biology, Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Nitra, Slovakia; Department of Molecular Biology and Genetics, Laboratory of Genome Dynamics and Stability, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Sofia, Bulgaria.

Department of Biophysics and Radiobiology, Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv, Ukraine.

出版信息

DNA Repair (Amst). 2017 Feb;50:14-21. doi: 10.1016/j.dnarep.2016.12.002. Epub 2016 Dec 16.

Abstract

This pilot study was carried out to assess the effect of radio-contaminated Chernobyl environment on plant genome integrity 27 years after the accident. For this purpose, nuclei were isolated from root tips of the soybean seedlings harvested from plants grown in the Chernobyl area for seven generations. Neutral, neutral-alkaline, and methylation-sensitive comet assays were performed to evaluate the induction and repair of primary DNA damage and the epigenetic contribution to stress adaptation mechanisms. An increased level of single and double strand breaks in the radio-contaminated Chernobyl seedlings at the stage of primary root development was detected in comparison to the controls. However, the kinetics of the recovery of DNA breaks of radio-contaminated Chernobyl samples revealed that lesions were efficiently repaired at the stage of cotyledon. Methylation-sensitive comet assay revealed comparable levels in the CCGG methylation pattern between control and radio-contaminated samples with a slight increase of approximately 10% in the latter ones. The obtained preliminary data allow us to speculate about the onset of mechanisms providing an adaptation potential to the accumulated internal irradiation after the Chernobyl accident. Despite the limitations of this study, we showed that comet assay is a sensitive and flexible technique which can be efficiently used for genotoxic screening of plant specimens in natural and human-made radio-contaminated areas, as well as for safety monitoring of agricultural products.

摘要

这项初步研究旨在评估切尔诺贝利事故27年后受放射性污染的环境对植物基因组完整性的影响。为此,从在切尔诺贝利地区种植了七代的大豆幼苗根尖中分离出细胞核。进行了中性、中性碱性和甲基化敏感彗星试验,以评估原发性DNA损伤的诱导和修复以及表观遗传对应激适应机制的贡献。与对照相比,在主根发育阶段,受切尔诺贝利放射性污染的幼苗中单链和双链断裂水平增加。然而,受切尔诺贝利放射性污染样本的DNA断裂恢复动力学表明,在子叶阶段损伤得到了有效修复。甲基化敏感彗星试验显示,对照样本和受放射性污染样本的CCGG甲基化模式水平相当,后者略有增加,约为10%。获得的初步数据使我们能够推测切尔诺贝利事故后为累积的内部辐射提供适应潜力的机制的启动。尽管本研究存在局限性,但我们表明彗星试验是一种灵敏且灵活的技术,可有效用于对天然和人为放射性污染地区的植物标本进行遗传毒性筛选,以及对农产品进行安全监测。

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