如何提高作物杂交种的生产效率是现代农业科技的重要目标之一. 目前, 玉米杂交制种主要采用母本物理去雄或胞质型不育系等方法, 制种质量控制难, 效率低且成本高昂. 细胞核基因不育系因其易于建立恢保关系而具有显著优势, 光温敏感型核不育基因已在水稻杂交种产业中实现大规模应用, 但在玉米种业上仍是尚未攻克的难题, 限制了杂交种生产的效率. 本研究采用CRISPR/Cas9基因编辑技术靶向编辑玉米ZmTms5基因, 创制了11种该基因功能丧失型突变体. 表型鉴定结果表明, 突变体在四分体至花粉成熟期日平均气温(day average temperatures, DAT)达28 ℃的高温条件下, 表现为雄性不育; 而在DAT不高于25 ℃的低温条件下, 花粉发育正常, 表现为雄性可育. 在全国玉米主要生态区域的14个试验地点进行的表型鉴定试验进一步证实了这一发现, 并明确了适宜繁亲和繁杂的区域. 本研究不仅提供了一种基于基因编辑的玉米温敏型核不育系创制技术策略, 而且为该类种质的杂交制种应用明确了产业化技术布局区域. 为提高杂交种制种效率和降低成本提供了新的解决方案, 具有重要的理论研究和产业应用价值. 此外, 本研究也为其它作物的温敏不育系创制提供了参考.

The Manipulated Genic Male Sterile Maintainer (MGM) system, a next-generation hybrid seed technology, enables efficient production of sortable seeds from genic male sterile (GMS) lines. However, implementing robust MGM systems in commercial maize inbred lines requires stable transformation, a genotype-specific and laborious process. This study aimed to integrate MGM technology into the commercial maize inbred line Z372, developing both GMS and MGM lines. We utilized the MGM line ZC01-3A-7, which contains the MS26ΔE5 editor T-DNA and MGM T-DNA, previously established in the highly transformable ZC01 recipient plants. Through a combination of crossing and backcrossing with Z372, we targeted the fertility gene Ms26 within the Z372 genome for mutation using the in vivo CRISPR/Cas9 activity within the MS26ΔE5 editor T-DNA construct. This approach facilitated precise editing of the Ms26 locus, minimizing linkage drag associated with the Ms26 mutation. Whole-genome SNP analysis achieved a 98.74% recovery rate for GMS and 96.32% for MGM in the BC2F2 generation. Importantly, the Z372-GMS line with the ms26ΔE5 mutation is non-transgenic, avoiding linkage drag and demonstrating production readiness. This study represents a significant advancement in maize breeding, enabling the rapid generation of GMS and MGM lines for efficient hybrid seed production.