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Generation of paternal haploids in wheat by genome editing of the centromeric histone CENH3

Nature Biotechnology volume 38, pages 1397–1401 (2020)Cite this article

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Abstract

New breeding technologies accelerate germplasm improvement and reduce the cost of goods in seed production1,2,3. Many such technologies could use in vivo paternal haploid induction (HI), which occurs when double fertilization precedes maternal (egg cell) genome loss. Engineering of the essential CENTROMERIC HISTONE (CENH3) gene induces paternal HI in Arabidopsis4,5,6. Despite conservation of CENH3 function across crops, CENH3-based HI has not been successful outside of the Arabidopsis model system7. Here we report a commercially operable paternal HI line in wheat with a ~7% HI rate, identified by screening genome-edited TaCENH3α-heteroallelic combinations. Unlike in Arabidopsis, edited alleles exhibited reduced transmission in female gametophytes, and heterozygous genotypes triggered higher HI rates than homozygous combinations. These developments might pave the way for the deployment of CENH3 HI technology in diverse crops.

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Fig. 1: Genome editing strategy and generation of HI alleles.
Fig. 2: Haploid and inducer phenotypes.

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Data availability

The authors declare that all data supporting the findings of this study are available in the manuscript and the Supplementary Materials.

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Acknowledgements

We thank Z. Chai, L. Geng, Y. Liu, X. Zhang, R. Cui and Y. Ren for their work generating the genome-edited plants. Thanks to Y. Ma and Y. Zhang for data analysis. We thank Y. Gao and W. Teng for wheat outcrossing and G. Tang for gene sequencing and characterization of CENH3 expression. Thanks to M. Zong for ploidy analysis support and to W. Huang, C. Ma, Y. Pan, X. He, W. Jin, C. Gao and F. Han for technique suggestions. We thank J. Green, L. Kavanaugh, M. Rose and X. Tan for bioinformatics support and C. Leming for intellectual property guidance. We thank X. Zhang, D. Skibbe, X. Chen, J. Xu, K. White, T. Zhu, W. Cao, R. Quadt, B. Anindya, L. Shi, E. Dunder, I. Jepsen, G. Wu and B. Zhang for leadership and project guidance.

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Author notes

  1. These authors contributed equally: Jian Lv, Kun Yu.

Authors and Affiliations

  1. Seeds Research, Syngenta Biotechnology China, ZhongGuanCun Life Science Park, Beijing, China

    Jian Lv, Kun Yu, Juan Wei, Huaping Gui, Chunxia Liu, Dawei Liang, Yanli Wang, Hongju Zhou, Tiancong Lu & Xingping Zhang

  2. Seeds Research, Syngenta Crop Protection, LLC, Research Triangle Park, NC, USA

    Ryan Carlin, Qiudeng Que, Wen Chung Wang & Tim Kelliher

  3. Seeds Research, Syngenta Seeds Research, Junction City, KS, USA

    Randy Rich

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  1. Jian Lv
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Contributions

J.L. and T.K. designed the experiments and wrote the manuscript. K.Y. managed plant care in the greenhouse, including seed increase and outcrossing. J.W. developed the genotyping strategy and conducted T2/F1 genotyping and KASP check. C.L. did the vector design and construction. H.G. and Y.W. performed the wheat transformation. H.Z. and J.L. sequenced T0/T1 genotypes. D.L. and J.L. performed pollen staining, stomata check and flower structure phenotyping, and R.C. performed the qRT–PCR of the three homoeologs of TaCENH3α and TaCENH3β in various tissues. T.L. analyzed histone modification sites. Q.Q., W.W., R.R. and X.Z. provided technique suggestions.

Corresponding authors

Correspondence to Jian Lv or Tim Kelliher.

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A patent covering the information in this manuscript was submitted on 2 October 2018.

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Supplementary Figs. 1–4 and Tables 1–7

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Lv, J., Yu, K., Wei, J. et al. Generation of paternal haploids in wheat by genome editing of the centromeric histone CENH3. Nat Biotechnol 38, 1397–1401 (2020). https://doi.org/10.1038/s41587-020-0728-4

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