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张雪, 张海波, 苏荣国, 汪靖超, 佘定懿, 宋修涵, 沙珍霞. 0#柴油对斑马鱼基因组DNA和环境DNA遗传毒性的RAPD比较[J]. 水生生物学报, 2020, 44(4): 764-773. DOI: 10.7541/2020.092
引用本文: 张雪, 张海波, 苏荣国, 汪靖超, 佘定懿, 宋修涵, 沙珍霞. 0#柴油对斑马鱼基因组DNA和环境DNA遗传毒性的RAPD比较[J]. 水生生物学报, 2020, 44(4): 764-773. DOI: 10.7541/2020.092
ZHANG Xue, ZHANG Hai-Bo, SU Rong-Guo, WANG Jing-Chao, SHE Ding-Yi, SONG Xiu-Han, SHA Zhen-Xia. COMPARISON OF DIESEL GENOTOXICITY ON ZEBRAFISH GENOMIC DNA AND ENVIRONMENTAL DNA FROM WATER ENVIRONMENT OF ZEBRAFISH BY RAPD[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(4): 764-773. DOI: 10.7541/2020.092
Citation: ZHANG Xue, ZHANG Hai-Bo, SU Rong-Guo, WANG Jing-Chao, SHE Ding-Yi, SONG Xiu-Han, SHA Zhen-Xia. COMPARISON OF DIESEL GENOTOXICITY ON ZEBRAFISH GENOMIC DNA AND ENVIRONMENTAL DNA FROM WATER ENVIRONMENT OF ZEBRAFISH BY RAPD[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(4): 764-773. DOI: 10.7541/2020.092

0#柴油对斑马鱼基因组DNA和环境DNA遗传毒性的RAPD比较

COMPARISON OF DIESEL GENOTOXICITY ON ZEBRAFISH GENOMIC DNA AND ENVIRONMENTAL DNA FROM WATER ENVIRONMENT OF ZEBRAFISH BY RAPD

  • 摘要: 研究利用随机扩增多态性DNA (Random Amplified Polymorphic DNA, RAPD)技术, 以斑马鱼基因组DNA和其养殖水体中的环境DNA (environmental DNA, eDNA)为模板, 检测0#柴油可溶性组分对斑马鱼(Danio rerio)遗传毒性的影响。结果显示, 通过基因组DNA和eDNA扩增的RAPD图谱均可检测到0#柴油对斑马鱼的遗传毒性。在未受到柴油暴露时, 斑马鱼基因组DNA和水环境中eDNA在96h内的RAPD图谱均无明显变化; 在不同浓度的柴油暴露下, 随着暴露时间(0、24h、48h、72h、96h)延长, 基因组DNA和eDNA的多态性位点减少, 模板稳定性降低; 随着柴油浓度(15%、50%、100%)的增加, 基因组DNA和eDNA的多态性位点也减少, 模板稳定性降低。这表明0#柴油对斑马鱼基因组DNA和eDNA的遗传毒性均呈现时间-效应和浓度-效应关系, 并且无论以斑马鱼基因组DNA还是eDNA为模板, 柴油暴露组和未进行暴露的对照组的RAPD扩增图谱条带变化趋势一致。研究结果为通过RAPD技术检测柴油对水生生物的遗传毒性提供了新的研究思路和技术手段。

     

    Abstract: The increased demand of diesel fuel enhanced the risk of diesel leaks causing deleterious effects on the aquatic environments. However, attempts to establish useful tools that use environmental DNA (eDNA) to detect genotoxicity by water environment contaminants have received little attention. The present study used RAPD (Random Amplified Polymorphic DNA) technology to detect the genotoxicity of soluble fraction of 0# diesel on the zebrafish (Danio rerio) using zebrafish genomic DNA and water environment DNA (eDNA) as templates. The results showed no significant change in the RAPD fingerprint of both genomic DNA and eDNA within 96h under control condition. When zebrafish were exposed to different concentration diesel with the prolonged exposure time (0, 24h, 48h, 72h, 96h), the polymorphic sites and template stability of genomic DNA and eDNA decreased. The increased diesel concentration (15%, 50%, 100%) reduced the polymorphic sites and the template stability of genomic DNA and eDNA. These results indicated that 0# diesel has a time- and concentration-dependent effects on the genotoxicity of zebrafish genomic DNA and eDNA. Moreover, the pattern of RAPD amplification fingerprint of genomic DNA was consistent with that of eDNA RAPD amplification fingerprint. Our research provides new research clues and technical means for detecting the genotoxicity of diesel oil on aquatic organisms by measuring the eDNA damage in water environment using RAPD technique.

     

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