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Expediting Next-Generation Hybrid Technology in Recalcitrant Maize Inbreds through In Vivo Targeted Activity of CRISPR/Cas9

文献类型： 外文期刊

作者： Hou, Liudi 1 ; Xiao, Bing 3 ; Zhu, Jinjie 2 ; Liu, Changlin 2 ; Wu, Qingyu 3 ; Xie, Chuanxiao 2 ; Zou, Huawen 1 ; Qi, Xiantao 2 ;

作者机构： 1.Yangtze Univ, Coll Agr, Jingzhou 434000, Peoples R China

2.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China

3.Chinese Acad Agr Sci, Inst Agr Resources & Reg Planning, State Key Lab Efficient Utilizat Arid & Semiarid A, Beijing 100081, Peoples R China

4.China Agr Univ, Natl Acad Agr Green Dev, Coll Resources & Environm Sci, Key Lab Plant Soil Interact,Minist Educ, Beijing 100193, Peoples R China

关键词： Zea mays L.; molecular breeding; next-generation hybrid technology; CRISPR/Cas9; genic male sterile

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：4.9； 五年影响因子：5.6 ）

ISSN： 1661-6596

年卷期： 2024 年 25 卷 11 期

页码：

收录情况： SCI

摘要： 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 MS26 Delta E5 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 MS26 Delta E5 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 ms26 Delta E5 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.

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