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Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis

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Abstract

DNA damage is the dominant source of mutation, which is the driving force of evolution. Therefore, it is important to quantitatively analyze the DNA damage caused by different mutagenesis methods, the subsequent mutation rates, and their relationship. Atmospheric and room temperature plasma (ARTP) mutagenesis has been used for the mutation breeding of more than 40 microorganisms. However, ARTP mutagenesis has not been quantitatively compared with conventional mutation methods. In this study, the umu test using a flow-cytometric analysis was developed to quantify the DNA damage in individual viable cells using Salmonella typhimurium NM2009 as the model strain and to determine the mutation rate. The newly developed method was used to evaluate four different mutagenesis systems: a new ARTP tool, ultraviolet radiation, 4-nitroquinoline-1-oxide (4-NQO), and N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) mutagenesis. The mutation rate was proportional to the corresponding SOS response induced by DNA damage. ARTP caused greater DNA damage to individual living cells than the other conventional mutagenesis methods, and the mutation rate was also higher. By quantitatively comparing the DNA damage and consequent mutation rate after different types of mutagenesis, we have shown that ARTP is a potentially powerful mutagenesis tool with which to improve the characteristics of microbial cell factories.

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Acknowledgments

This work was supported by the Tsinghua University Initiative Scientific Research Program (2011THZ 01019) and the JST CREST project of Japan.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical Engineering, Tsinghua University, Beijing, China

    Xue Zhang, Chong Zhang, Li-Yan Wang, Hai-Bo Chang & Xin-Hui Xing

  2. Biobreeding Research Center, Wuxi Research Institute of Applied Technologies, Tsinghua University, Wuxi, Jiangsu, China

    Chong Zhang, Qian-Qian Zhou, Li-Yan Wang, He-Ping Li & Xin-Hui Xing

  3. Department of Engineering Physics, Tsinghua University, Beijing, China

    Xiao-Fei Zhang & He-Ping Li

  4. Institute of Life and Environmental Sciences, Osaka Shin-Ai College, Tsurumi, Tsurumi-ku, Osaka, Japan

    Yoshimitsu Oda

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  1. Xue Zhang
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  2. Chong Zhang
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  3. Qian-Qian Zhou
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  9. Xin-Hui Xing
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Correspondence to Chong Zhang or Xin-Hui Xing.

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Zhang, X., Zhang, C., Zhou, QQ. et al. Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis. Appl Microbiol Biotechnol 99, 5639–5646 (2015). https://doi.org/10.1007/s00253-015-6678-y

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