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基因敲除是植物功能基因组研究和农艺性状定向遗传改良的关键步骤,近年来,CRISPR/Cas9介导的目标基因敲除在拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)等植物中取得飞速的发展,然而在棉花(Gossypium spp.)中目前尚没有成功的报道.陆地棉(G.hirsutum L.)是典型的异源四倍体模式作物,包括A亚基因组和D亚基因组.利用CRISPR/Cas9技术同时成功地敲除A和D亚基因组精氨酸酶编码基因(GhARG),Gh_A05G2143和Gh_D05G2397.无论高氮还是低氮条件下,T1代双基因纯和株系侧根生长发育显著地提高.因此,本研究的CRISPR/Cas9系统可以同时高效地编辑棉花中多个同源基因,从而实现关键农艺性状简单快速得改良.
In recent years, CRISPR / Cas9-mediated knockdown of target genes has achieved rapid progress in plants such as Arabidopsis thaliana and Oryza sativa However, there are no successful reports in cotton (Gossypium spp.) G. hirsutum L. is a typical allotetraploid crop including A subgenome and D subgenome.Using CRISPR / Cas9 knock-out of both A and D subunit genomic arginase-encoding genes (GhARG), Gh_A05G2143 and Gh_D05G2397. The growth and development of lateral roots of both pure lines and T1 lines were significantly increased under both high and low N levels Therefore, the CRISPR / Cas9 system of this study can edit multiple homologous genes in cotton efficiently at the same time, so that the key agronomic traits can be quickly and easily improved.