Genome-Edited Triple-Recessive Mutation Alters Seed Dormancy in Wheat
Genome-Edited Triple-Recessive Mutation Alters Seed Dormancy in Wheat
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DOI：
10.1016/j.celrep.2019.06.090
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发表时间：
2019-07-30
期刊：
影响因子：
8.8
通讯作者：
Sato, Kazuhiro
中科院分区：
文献类型：
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作者：
Abe, Fumitaka;Haque, Emdadul;Sato, Kazuhiro
普通小麦有三套亚基因组，使得突变难以观察，特别是对于由隐性基因控制的性状。在这里，我们通过农杆菌介导的CRISPR/Cas9产生了Qsd 1同源等位基因功能丧失的六倍体小麦品系，Qsd 1控制大麦种子休眠。在产生的8个转化小麦事件中，获得了3个携带小麦Qsd 1同源等位基因中的多个突变的独立事件。值得注意的是，一个品系在每个同源等位基因中都有突变。我们将这种植物与野生型栽培品种菲尔德杂交，产生一个无转基因的三隐性突变体，如孟德尔分离所揭示的。该突变体种子休眠期明显长于野生型，这可能导致穗上籽粒收获前发芽减少。PCR、Southern印迹和全基因组鸟枪测序表明，该分离株的基因组序列中缺乏转基因。该技术可作为小麦性状改良的模型，特别是遗传隐性性状，基于来自二倍体大麦的基因座信息。
Common wheat has three sets of sub-genomes, making mutations difficult to observe, especially for traits controlled by recessive genes. Here, we produced hexaploid wheat lines with loss of function of homeoalleles of Qsd1, which controls seed dormancy in barley, by Agrobacterium-mediated CRISPR/Cas9. Of the eight transformed wheat events produced, three independent events carrying multiple mutations in wheat Qsd1 homeoalleles were obtained. Notably, one line had mutations in every homeoallele. We crossed this plant with wild-type cultivar Fielder to generate a transgene-free triple-recessive mutant, as revealed by Mendelian segregation. The mutant showed a significantly longer seed dormancy period than wild-type, which may result in reduced pre-harvest sprouting of grains on spikes. PCR, southern blotting, and whole-genome shotgun sequencing revealed that this segregant lacked transgenes in its genomic sequence. This technique serves as a model for trait improvement in wheat, particularly for genetically recessive traits, based on locus information from diploid barley.