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CRISPR/Cas9-mediated editing of PhMLO1 confers powdery mildew resistance in petunia

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Abstract

Mildew Locus O (MLO), which encodes a seven-transmembrane domain protein, is responsible for the development of powdery mildew disease symptoms in many plant species. Hence, we knocked out PhMLO1 from Petunia hybrida cv. Mirage Rose at the genomic level using the Clustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) system and investigated involvement of the gene in powdery mildew disease development in the petunia. The MLO1 transcript levels observed in all mlo1 mutants were significantly lower than that in the wild type (WT). Specifically, the transcript level was the lowest in the mutants (line no. 1, 25, 33, and 85), followed by the mutants (line no. 14, 17, and 81), and line no. 6. Disease development was positively linked to the higher transcript levels of PhMLO1, because the disease severity observed in WT was greater than that observed in the mutants (line no. 6,14, 17, and 81), and disease symptoms were almost not observed in the mutants (line no. 1, 25, 33, and 85). In addition, the transmission of the edited alleles from the T0 to T1 generation was also confirmed, and the disease severity results were identical to those observed in the T0 mutant lines. Overall, the editing of PhMLO1 completely or partially prevented the development of this disease in this petunia, depending on MLO1 transcript level of the mutants. This study highlights the role of PhMLO1 in the development of powdery mildew disease in petunia and points to the need to edit the MLO genes in other ornamental plants to improve ornamental quality against powdery mildew disease.

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Acknowledgements

This work was carried out with the support of "Cooperative Research Program for Agriculture Science & Technology Development  (Project No. RS-2022-RD010015)" Rural Development Administration, Republic of Korea.

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Author notes

  1. Junping Xu and Aung Htay Naing have equally contributed to this work.

Authors and Affiliations

  1. Department of Horticultural Science, Kyungpook National University, Daegu, 41566, South Korea

    Junping Xu, Aung Htay Naing, Hyunhee Kang & Chang Kil Kim

  2. Life Science and Technology School, Lingnan Normal University, Zhanjiang, 524048, China

    Junping Xu

  3. Floriculture Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju, 55365, South Korea

    Su Young Lee

  4. National Agrobiodiversity Center, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju, 54874, Republic of Korea

    Weilan Li

  5. Department of Agricultural Education, Sunchon National University, Suncheon, South Korea

    Mi Young Chung

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  1. Junping Xu
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Contributions

AHN and JX designed the study. WL constructed the vector and JX assisted the vector construction. MYC generated MLO1-edited transgenic lines. JX extracted DNA for Sanger sequencing and did powdery mildew test. JX produced T1 mutants and did powdery mildew test. AHN and JX analyzed the data. HK assisted the experiments. SYL helped fungal infection in the mutants. AHN wrote and revised the manuscript. CKK supervised the project. All authors read and approved the final manuscript.

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Correspondence to Aung Htay Naing or Chang Kil Kim.

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Xu, J., Naing, A.H., Kang, H. et al. CRISPR/Cas9-mediated editing of PhMLO1 confers powdery mildew resistance in petunia. Plant Biotechnol Rep 17, 767–775 (2023). https://doi.org/10.1007/s11816-023-00854-5

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  • DOI: https://doi.org/10.1007/s11816-023-00854-5

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