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A DMP-triggered in vivo maternal haploid induction system in the dicotyledonous Arabidopsis

Abstract

Doubled haploid technology using inducer lines carrying mutations in ZmPLA1/MTL/NLD and ZmDMP1,2,3,4 has revolutionized traditional maize breeding. ZmPLA1/MTL/NLD is conserved in monocots and has been used to extend the system from maize to other monocots5,6,7, but no functional orthologue has been identified in dicots, while ZmDMP-like genes exist in both monocots and dicots4,8,9. Here, we report that loss-of-function mutations in the Arabidopsis thaliana ZmDMP-like genes AtDMP8 and AtDMP9 induce maternal haploids, with an average haploid induction rate of 2.1 ± 1.1%. In addition, to facilitate haploid seed identification in dicots, we established an efficient FAST-Red fluorescent marker-based haploid identification system that enables the identification of haploid seeds with >90% accuracy. These results show that mutations in DMP genes also trigger haploid induction in dicots. The conserved expression patterns and amino acid sequences of ZmDMP-like genes in dicots suggest that DMP mutations could be used to develop in vivo haploid induction systems in dicots.

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Fig. 1: Mutation of Arabidopsis DMP genes results in multiple seed phenotypes.
Fig. 2: Haploid seed production and identification with the FAST-Red marker.

Data availability

The datasets generated and/or analysed during the current study are available from the corresponding author on request. The raw whole-genome sequencing data have been deposited in the NCBI Sequence Read Archive under the accession code PRJNA608056. Source Data for Figs. 1 and 2 are provided with the paper.

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Acknowledgements

We thank D. Ye and L. Chen for providing the ms1 and ms1-1 mutants, X. Yang for help with the analysis of haploid sequence data, Y. Leng for help with the analysis of seed development, and Z. Li for help with the phylogenetic analysis. This work was supported by the National Key Research and Development Program of China (2016YFD0101200 and 2018YFD0100201), Modern Maize Industry Technology System (CARS-02-04) and National Natural Science Foundation of China (91935303) (to S.C), as well as a China Scholarship Council grant (to B.C).

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Authors and Affiliations

Authors

Contributions

Y.Z., B.C., C.L. and S.C. conceived of and designed the experiments. Y.Z., M.L. and B.C. performed most of the experiments. D.W., Y.J., X.Q., Z.L., C.C., Y.W., M.C., J.L., Z.X. and D.C. performed some of the experiments. Y.Z., B.C., S.C., C.L., M.W., W.L. and M.L. analysed the data. Y.Z., B.C., C.L., K.B. and S.C. discussed and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Chenxu Liu or Shaojiang Chen.

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The authors declare no competing interests.

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Peer review information Nature Plants thanks Thomas Widiez and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–6 and Tables 2, 4, 6 and 7.

Reporting Summary

Supplementary Table 1

ZmDMP-like genes in plants.

Supplementary Table 3

A summary of CRISPR–Cas9 mutagenesis.

Supplementary Table 5

Genotyping data of haploids.

Supplementary Table 8

SNP data of haploids.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

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Zhong, Y., Chen, B., Li, M. et al. A DMP-triggered in vivo maternal haploid induction system in the dicotyledonous Arabidopsis. Nat. Plants 6, 466–472 (2020). https://doi.org/10.1038/s41477-020-0658-7

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