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Enhanced rice salinity tolerance via CRISPR/Cas9-targeted mutagenesis of the OsRR22 gene

机译:通过osrr22基因的CRISPR / CAS9靶向诱变增强水稻盐度耐受性

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Salinity is one of the most important abiotic stress affecting the world rice production. The cultivation of salinity-tolerant cultivars is the most cost-effective and environmentally friendly approach for salinity control. In recent years, CRISPR/Cas9 systems have been widely used for target-site genome editing; however, their application for the improvement of elite rice cultivars has rarely been reported. Here, we report the improvement of the rice salinity tolerance by engineering a Cas9-OsRR22-gRNA expressing vector, targeting the OsRR22 gene in rice. Nine mutant plants were identified from 14 T-0 transgenic plants. Sequencing showed that these plants had six mutation types at the target site, all of which were successfully transmitted to the next generations. Mutant plants without transferred DNA (T-DNA) were obtained via segregation in the T1 generations. Two T2 homozygous mutant lines were further examined for their salinity tolerance and agronomic traits. The results showed that, at the seedling stage, the salinity tolerance of T2 homozygous mutant lines was significantly enhanced compared to wild-type plants. Furthermore, no significantly different agronomic traits were found between T2 homozygous mutant lines and wild-type plants. Our results indicate CRISPR/Cas9 as a useful approach to enhance the salinity tolerance of rice.
机译:盐度是影响世界稻米生产的最重要的非生物压力之一。盐度耐受栽培品种是最具成本效益和环保的盐度控制的方法。近年来,CRISPR / CAS9系统已广泛用于靶网站基因组编辑;然而,他们对改善精英水稻品种的应用很少。在这里,我们通过工程通过工程CAS9-OSRR22-GRNA表达载体来提高水稻盐度耐受性,靶向水稻中的OSRR22基因。从14个T-0转基因植物鉴定九种突变植物。测序表明,这些植物在靶位点具有六种突变类型,所有这些植物成功地传播到下一代。通过T1代中的偏析获得不转移DNA(T-DNA)的突变植物。进一步检查两种T2纯合突变体,用于它们的盐度耐受性和农艺性状。结果表明,与野生型植物相比,在幼苗阶段,T2纯合突变体系的盐度耐受性显着增强。此外,在T2纯合突变体系和野生型植物之间发现了没有显着不同的农艺性状。我们的结果表明CRISPR / CAS9作为提高水稻盐度耐受性的有用方法。

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