Haploid induction (HI) is an important tool in crop breeding. Phospholipase A1 (ZmPLA1)/NOT LIKE DAD (NLD)/MATRILINEAL (MTL) is a key gene controlling HI in maize; however, the underlying molecular mechanism remains unclear. In this study, to dissect why loss of ZmPLA1 function could mediate HI we performed a comprehensive multiple omics analysis of zmpla1 mutant anthers by integrating transcriptome, metabolome, quantitative proteome, and protein modification data. Functional classes of significantly enriched or differentially abundant molecular entities were found to be associated with the oxidative stress response, suggesting that a reactive oxygen species (ROS) burst plays a critical role in HI. In support of this, we further discovered that a simple chemical treatment of pollen with ROS reagents could lead to HI. Moreover, we identified ZmPOD65, which encodes a sperm-specific peroxidase, as a new gene controlling HI. Taken together, our study revealed a likely mechanism of HI, discovered a new gene controlling HI, and created a new method for HI in maize, indicating the importance of ROS balance in maintaining normal reproduction and providing a potential route to accelerate crop breeding.

单倍体诱导（HI）是作物育种的重要工具。磷脂酶A1（ZmPLA1）/NOT LIKE DAD（NLD）/MATRILINEAL（MTL）是控制玉米HI的关键基因；然而，潜在的分子机制仍不清楚。在这项研究中，为了剖析为什么ZmPLA1功能的丧失可以介导 HI，我们对zmpla1进行了全面的多组学分析通过整合转录组、代谢组、定量蛋白质组和蛋白质修饰数据来获得突变花药。发现显着富集或差异丰富的分子实体的功能类别与氧化应激反应有关，这表明活性氧 (ROS) 爆发在 HI 中起关键作用。为了支持这一点，我们进一步发现用 ROS 试剂对花粉进行简单的化学处理可能会导致 HI。此外，我们确定了 ZmPOD65，它编码精子特异性过氧化物酶，作为控制 HI 的新基因。综上所述，我们的研究揭示了 HI 的可能机制，发现了控制 HI 的新基因，并创造了玉米 HI 的新方法，表明 ROS 平衡在维持正常繁殖中的重要性，并为加速作物育种提供了潜在途径。