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Genome Editing Enables Next-Generation Hybrid Seed Production Technology.
Molecular Plant ( IF 27.5 ) Pub Date : 2020-07-08 , DOI: 10.1016/j.molp.2020.06.003
Xiantao Qi 1 , Congsheng Zhang 1 , Jinjie Zhu 1 , Changlin Liu 1 , Changling Huang 1 , Xinhai Li 2 , Chuanxiao Xie 1
Affiliation  

The next-generation hybrid seed technology enables the successful production of sortable hybrid seeds from genic male sterile (GMS) lines and maintainers; however, it requires multiple laborious and complicated steps. Here, we designed a simple next-generation hybrid seed production strategy that takes advantage of the CRISPR/Cas9 technology to create a Manipulated GMS Maintainer (MGM) system via a single transformation. Under this schema, the maize male fertility gene ZmMS26 was nullified by removal of its fifth exon using the CRISPR/Cas9 system on a vector, and a second vector carrying a functional ZmMS26 cDNA was co-transformed to restore fertility. The second vector also contains a male gametophyte inactivation gene (ZmAA1) encoding maize α-amylase driven by the pollen-specific promoter PG47 and an endosperm fluorescent marker (DsRED) driven by the barley endosperm aleurone-specific promoter Ltp2. The derived single-copy hemizygous MGM lines bore a mutated MS26 gene, leading to complete male sterility but normal vegetative growth and grain yield. The MGM system could prevent genetic transmission of the MGM elements via male gametophytes, providing an efficient method for sorting maintainer seeds labeled by DsRED. This strategy can be extended to any GMS gene and to hybrid crops other than maize.



中文翻译:

基因组编辑启用了下一代杂交种子生产技术。

下一代杂交种子技术可从基因雄性不育(GMS)品系和维持者成功生产可分选的杂交种子。然而,这需要多个费力且复杂的步骤。在这里,我们设计了一种简单的下一代杂交种子生产策略,该策略利用CRISPR / Cas9技术通过一次转化创建了一个可操纵的GMS保持器(MGM)系统。在这种模式下,通过在载体上使用CRISPR / Cas9系统去除其第五个外显子,使玉米雄性育性基因ZmMS26无效,并将带有功能性ZmMS26 cDNA的第二个载体共转化以恢复生育力。第二个载体还包含一个雄配子体失活基因(ZmAA1)编码花粉特异性启动子PG47驱动的玉米α-淀粉酶 和大麦胚乳糊粉特异性启动子Ltp2驱动的胚乳荧光标记(DsRED)。衍生的单拷贝半合子MGM品系带有突变的MS26基因,导致完全雄性不育,但营养生长和谷物产量正常。MGM系统可以阻止MGM元素通过雄配子体的遗传传播,从而为分选DsRED标记的维持种子提供了一种有效的方法。该策略可以扩展到任何GMS基因,也可以扩展到玉米以外的杂交作物。

更新日期:2020-07-08
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