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CRISPR/Cas9 gene editing in peanut by Agrobacterium tumefaciens-mediated pollen tube transformation

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Abstract

Peanut (Arachis hypogaea L.) is an important global food crop, providing a valuable source of protein, oil and nutrition. The functional analysis of genes associated with yield traits is an important step in molecular breeding, which requires effective methods of genetic transformation. In this study, we applied the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 system in conjunction with Agrobacterium tumefaciens-mediated pollen tube transformation to create knockout mutants of the peanut MUTATOR-LIKE TRANSPOSABLE ELEMENT9A (AhMULE9A) gene, which plays an importantly regulatory role in response to Al stress in peanut. The gene mutations were identified by the analysis of genomic sequences flanking the sgRNA target sites. The sequencing of AhMULE9A mutations showed that eleven of the fifteen transgenics produced resulted in protein sequence changes, demonstrating the efficacy of this method for the production of targeted mutations in allotetraploid peanut.

Key message

This study highlights the efficiency and utility of CRISPR/Cas9 gene editing in peanut in conjunction with Agrobacterium tumefaciens-mediated pollen tube transformation.

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Data sharing not applicable to this article as no datasets were generated or analyzed during the current study.

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Acknowledgements

This work was financially supported by the Central Guidance for Local Science and Technology Development Funds Projects (Grant No. ZY22096013), the Guangxi Natural Science Foundation (Grant No. 2022GXNSFAA035437), the Guangxi Innovation Team Project of Soybean and Oil Crops of Modern Agricultural Industrial Technology System of China (nycytxgxcxtd-20-02) and Training Program for 1000 Young and Middle-aged Key Teachers in Guangxi at 2019.

Funding

This study was funded by the Central Guidance for Local Science and Technology Development Funds Projects (Grant No. ZY22096013), the Guangxi Natural Science Foundation (Grant No. 2022GXNSFAA035437), the Guangxi Innovation Team Project of Soybean and Oil Crops of Modern Agricultural Industrial Technology System of China (nycytxgxcxtd-20-02) and Training Program for 1000 Young and Middle–aged Key Teachers in Guangxi at 2019.

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Authors and Affiliations

  1. College of Agriculture, Guangxi University, Nanning, 530004, Guangxi, China

    Ailing Li, Min Zhou, Guoting Liao, Xinyue Li, Aiqin Wang, Dong Xiao, Longfei He & Jie Zhan

  2. Key Laboratory of Crop Cultivation and Physiology, Education Department of Guangxi Zhuang Autonomous Region, Guangxi University, Nanning, 530004, China

    Aiqin Wang, Dong Xiao, Longfei He & Jie Zhan

  3. Guangxi Key Laboratory of Agro-environment and Agric-products Safety, College of Agriculture, Guangxi University, Nanning, 530004, Guangxi, China

    Aiqin Wang, Dong Xiao, Longfei He & Jie Zhan

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  1. Ailing Li
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  2. Min Zhou
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  3. Guoting Liao
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  4. Xinyue Li
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  8. Jie Zhan
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Contributions

JZ conceived the experiments and revised this manuscript; ALL performed most of the experiments and wrote the manuscript; MZ, GTL, XYL performed some of the experiments; AQW, DX, LFH and JZ analyzed the data. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Jie Zhan.

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Communicated by Sergio J. Ochatt.

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Li, A., Zhou, M., Liao, G. et al. CRISPR/Cas9 gene editing in peanut by Agrobacterium tumefaciens-mediated pollen tube transformation. Plant Cell Tiss Organ Cult 155, 883–892 (2023). https://doi.org/10.1007/s11240-023-02607-2

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