Mutation of ZmDMP enhances haploid induction in maize

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Abstract

Doubled haploid (DH) breeding based on in vivo haploid induction has led to a new approach for maize breeding1. All modern haploid inducers used in DH breeding are derived from the haploid inducer line Stock6. Two key quantitative trait loci, qhir1 and qhir8, lead to high-frequency haploid induction2. Mutation of the gene MTL/ZmPLA1/NLD in qhir1 could generate a ~2% haploid induction rate (HIR)3,4,5; nevertheless, this mutation is insufficient for modern haploid inducers whose average HIR is ~10%6. Therefore, cloning of the gene underlying qhir8 is important for illuminating the genetic basis of haploid induction. Here, we present the discovery that mutation of a non-Stock6-originating gene in qhir8, namely, ZmDMP, enhances and triggers haploid induction. ZmDMP was identified by map-based cloning and further verified by CRISPR–Cas9-mediated knockout experiments. A single-nucleotide change in ZmDMP leads to a 2–3-fold increase in the HIR. ZmDMP knockout triggered haploid induction with a HIR of 0.1–0.3% and exhibited a greater ability to increase the HIR by 5–6-fold in the presence of mtl/zmpla1/nld. ZmDMP was highly expressed during the late stage of pollen development and localized to the plasma membrane. These findings provide important approaches for studying the molecular mechanism of haploid induction and improving DH breeding efficiency in maize.

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Fig. 1: Map-based cloning of ZmDMP and knockout experiments.
Fig. 2: Phenotypic evidence of transgenic events that enhanced HIRs.
Fig. 3: Verification of the haploid phenotypes.
Fig. 4: Expression and subcellular localization profiling of ZmDMP and zmdmp.

Data availability

The data sets generated and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We acknowledge support from the National Key Research and Development Program of China—Maize heterosis utilization technology and strong heterosis hybrids breeding (2016YFD0101200, 2016YFD0101003 and 2018YFD0100201)—and the Modern Maize Industry Technology System (CARS–02–04).

Author information

S.C., Y.Z. and C.L. conceived and designed the experiments. Y.Z., C.L., X.Q. and Y.J. performed the experiments. Y.Z., S.C., C.L. and X.Q. analysed the data. C.L., S.C., Y.Z., D.W., Y.W., Z.L., C.C., B.C. X.T., J.Li, M.C., X.D., X.X., L.L., W.Li., W.Liu., W.J. and J.Lai. wrote the manuscript with input from all authors.

Correspondence to Shaojiang Chen.

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The authors declare no competing interests.

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Peer review information: Nature Plants thanks Jose Seguí-Simarro and other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Zhong, Y., Liu, C., Qi, X. et al. Mutation of ZmDMP enhances haploid induction in maize. Nat. Plants 5, 575–580 (2019) doi:10.1038/s41477-019-0443-7

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