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<p>Doubled haploid (DH) technology rapidly generates completely homozygous lines and is widely used in modern crop breeding. In <i>Brassica</i> crops such as Pak–choi (<i>Brassica rapa</i> ssp. <i>chinensis</i>), traditional DH methods, such as microspore culture, remain highly genotype–dependent and technically challenging due to whole–genome triplication, gene redundancy, and low regeneration efficiency. Consequently, the development of <i>in planta</i> genome–editing strategies is vital for overcoming these limitations and providing a practical alternative to conventional tissue culture–based approaches. Here, we review CRISPR/Cas9–mediated haploid induction in plants, focusing on three pathways: centromere–mediated genome elimination by modifying the centromere–specific histone <i>CENH3</i>, fertilization–related maternal haploid induction via the gynoecium–expressed patatin–related phospholipase <i>pPLAIIγ</i>, and male gametophyte–dependent haploid induction controlled by <i>DMP</i> genes. We compare these pathways regarding their molecular mechanisms, developmental timing, gene essentiality, and suitability for genome editing, with particular emphasis on <i>Brassica</i> species. Special attention is given to haploid inducer gene homologs in Pak–choi and to the challenges imposed by ancestral genome triplication in <i>Brassica rapa</i>. Finally, we discuss how to use CRISPR/Cas9–based haploid induction systems in <i>Brassica</i> breeding programs, highlighting their value for accelerating doubled haploid production and functional genomics in genetically complex crop species.</p>

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CRISPR/Cas9–mediated haploid induction in Brassica crops: a review of CENH3, pPLAIIγ, and DMP targets and breeding prospects

  • 期刊论文

  • 发表日期：

    2026年6月19日

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  • May Phyo Thu,
  • Young–Cheon Kim,
  • Jeong Hwan Lee

摘要

Doubled haploid (DH) technology rapidly generates completely homozygous lines and is widely used in modern crop breeding. In Brassica crops such as Pak–choi (Brassica rapa ssp. chinensis), traditional DH methods, such as microspore culture, remain highly genotype–dependent and technically challenging due to whole–genome triplication, gene redundancy, and low regeneration efficiency. Consequently, the development of in planta genome–editing strategies is vital for overcoming these limitations and providing a practical alternative to conventional tissue culture–based approaches. Here, we review CRISPR/Cas9–mediated haploid induction in plants, focusing on three pathways: centromere–mediated genome elimination by modifying the centromere–specific histone CENH3, fertilization–related maternal haploid induction via the gynoecium–expressed patatin–related phospholipase pPLAIIγ, and male gametophyte–dependent haploid induction controlled by DMP genes. We compare these pathways regarding their molecular mechanisms, developmental timing, gene essentiality, and suitability for genome editing, with particular emphasis on Brassica species. Special attention is given to haploid inducer gene homologs in Pak–choi and to the challenges imposed by ancestral genome triplication in Brassica rapa. Finally, we discuss how to use CRISPR/Cas9–based haploid induction systems in Brassica breeding programs, highlighting their value for accelerating doubled haploid production and functional genomics in genetically complex crop species.