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Genetic Manipulation of Reactive Oxygen Species (ROS) Homeostasis Utilizing CRISPR/Cas9-Based Gene Editing in Rice

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Reactive Oxygen Species in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2526))

Abstract

Reactive oxygen species (ROS) are now recognized as key signals in plant stress responses. Adverse environmental conditions can either promote ROS production or downregulate antioxidative enzymes, leading to the alteration of redox homeostasis and activation of ROS-linked stress signaling. To uncover their signaling mechanisms and to characterize related components, genetic modification of ROS homeostasis is a central approach. CRISPR/Cas9-based genome editing system has become a powerful tool for gene mutation in a variety of organisms, including plants. Within this chapter, we describe a method that can be applied to manipulate ROS homeostasis in rice (Oryza sativa L.) utilizing CRISPR/Cas9 technology. Step-by-step protocols including the design and construction of Cas9/sgRNA, agrobacterium-mediated transformation, and mutation characterization are described. Application of this system in editing a rice catalase gene CatC, a key antioxidative enzyme in controlling ROS homeostasis, is also presented.

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Acknowledgments

We would like to thank Prof. Yaoguang Liu (South China Agricultural University) for providing the original CRISPR/Cas9 vectors. This work was funded by grants from the National Natural Science Foundation of China (31300225, 31301798, 31822047) and start-up funding of Anhui Agricultural University.

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Authors and Affiliations

  1. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, Jiangsu, China

    Sheng Xu & Ren Wang

  2. School of Food and Biological Engineering, Hefei University of Technology, Hefei, Anhui, China

    Tao Chen, Mimi Tian, Linlin Zhang & Yi Han

  3. Institute of Ecology and Environmental Sciences of Paris, Molecular Ecophysiology of Stressed plant, Faculté des Sciences et Technologie, Université Paris Est Créteil, Créteil, France

    Marie-Sylviane Rahantaniaina

  4. MOA Key Laboratory of Plant Nutrition and Fertilization in Lower-Middle Reaches of the Yangtze River and State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, Jiangsu, China

    Wei Xuan

  5. National Engineering Laboratory of Crop Stress Resistance Breeding, School of Life Sciences, Anhui Agricultural University, Hefei, China

    Yi Han

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  1. Sheng Xu
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  2. Tao Chen
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  3. Mimi Tian
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  4. Marie-Sylviane Rahantaniaina
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  5. Linlin Zhang
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  6. Ren Wang
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  7. Wei Xuan
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  8. Yi Han
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Corresponding author

Correspondence to Yi Han .

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Editors and Affiliations

  1. VIB Center for Plant Systems Biology, Ghent University, Zwijnaarde, Belgium

    Amna Mhamdi

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Xu, S. et al. (2022). Genetic Manipulation of Reactive Oxygen Species (ROS) Homeostasis Utilizing CRISPR/Cas9-Based Gene Editing in Rice. In: Mhamdi, A. (eds) Reactive Oxygen Species in Plants. Methods in Molecular Biology, vol 2526. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2469-2_3

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  • DOI: https://doi.org/10.1007/978-1-0716-2469-2_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2468-5

  • Online ISBN: 978-1-0716-2469-2

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