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CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China

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Abstract

Key Message

The blast resistance allele of OsBsr-d1 does not exist in most japonica rice varieties of Jilin Province in China. The development of Bsr-d1 knockout mutants via CRISPR/Cas9 enhances broad-spectrum resistance to rice blast in Northeast China.

Abstract

Rice blast is a global disease that has a significant negative impact on rice yield and quality. Due to the complexity and variability of the physiological races of rice blast, controlling rice blast is challenging in agricultural production. Bsr-d1, a negative transcription factor that confers broad-spectrum resistance to rice blast, was identified in the indica rice cultivar Digu; however, its biological function in japonica rice varieties is still unclear. In this study, we analyzed the blast resistance allele of Bsr-d1 in a total of 256 japonica rice varieties from Jilin Province in Northeast China and found that this allele was not present in these varieties. Therefore, we generated Bsr-d1 knockout mutants via the CRISPR/Cas9 system using the japonica rice variety Jigeng88 (JG88) as a recipient variety. Compared with those of the wild-type JG88, the homozygous Bsr-d1 mutant lines KO#1 and KO#2 showed enhanced leaf blast resistance at the seedling stage to several Magnaporthe oryzae (M. oryzae) races collected from Jilin Province in Northeast China. Physiological and biochemical indices revealed that the homozygous mutant lines produced more hydrogen peroxide than did JG88 plants when infected with M. oryzae. Comparative RNA-seq revealed that the DEGs were mainly involved in the synthesis of amide compounds, zinc finger proteins, transmembrane transporters, etc. In summary, our results indicate that the development of Bsr-d1 knockout mutants through CRISPR/Cas9 can enhance the broad-spectrum resistance of rice in Northeast China to rice blast. This study not only provides a theoretical basis for disease resistance breeding involving the Bsr-d1 gene in Northeast China, but also provides new germplasm resources for disease-resistance rice breeding.

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The original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding author.

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Acknowledgements

This research was supported by grants from the Jilin Provincial Natural Science Fundation in China (20210101026JC) and the Jilin Provincial Agricultural Science and Technology Innovation Project in China (CXGC2023RCY050).

Funding

Natural Science Fundation of Jilin Province, 20210101026JC, Yong-Mei Jin, Jilin Provincial Agricultural Science and Technology Independent Innovation Fund, CXGC2023RCY050, Anqi Song.

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Authors and Affiliations

  1. Research Institute of Agricultural Biotechnology/Jilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, Changchun, 130033, People’s Republic of China

    Ying Zhang, Xiu-Feng Lin, Anqi Song, Mengmeng Gao & Yong-Mei Jin

  2. Rice Research Institute, Jilin Academy of Agricultural Sciences, Gongzhuling, 136100, People’s Republic of China

    Ying Zhang & Ri-Hua Piao

  3. Institute of Plant Protection, Jilin Academy of Agricultural Sciences/Jilin Key Laboratory of Agricultural Microbiology/Key Laboratory of Integrated Pest Management On Crops in Northeast China, Ministry of Agriculture and Rural Affairs, Gongzhuling, 136100, People’s Republic of China

    Li Li

  4. College of Agricultural Sciences, Yanbian University, Yanji, 133000, People’s Republic of China

    Songquan Wu, Anqi Song & Mengmeng Gao

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  1. Ying Zhang
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Contributions

Y-MJ and YZ designed the experiments. YZ, X-FL, LL, R-HP, SW, AS and MG performed the experiments. YZ, LL, R-HP and Y-MJ analyzed the data. Y-MJ, YZ and R-HP participated in the writing of the manuscript. All the authors contributed to the article and approved the submitted version.

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Correspondence to Yong-Mei Jin.

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Communicated by Zheng-Yi Xu.

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Zhang, Y., Lin, XF., Li, L. et al. CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China. Plant Cell Rep 43, 100 (2024). https://doi.org/10.1007/s00299-024-03192-0

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