Background

Genome editing tools are important for functional genomics research and biotechnology applications. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 (Cas9) system for gene knockout has emerged as the most effective genome-editing tool. It has previously been reported that, in rice plants, knockdown of the Os8N3 gene resulted in enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo), while displaying abnormal pollen development.

Results

The CRISPR/Cas9 system was employed to knockout rice Os8N3, in order to confer enhanced resistance to Xoo. Analysis of the genotypes and edited Os8N3 in T₀, T₁, T₂, and T₃ transgenic rice plants showed that the mutations were transmitted to subsequent generations, and homozygous mutants displayed significantly enhanced resistance to Xoo. Stable transmission of CRISPR/Cas9-mediated Os8N3 gene editing without the transferred DNA (T-DNA) was confirmed by segregation in the T₁ generation. With respect to many investigated agronomic traits including pollen development, there was no significant difference between homozygous mutants and non-transgenic control plants under greenhouse growth conditions.

Conclusion

Data from this study indicate that the CRISPR/Cas9-mediated Os8N3 edition can be successfully employed for non-transgenic crop improvements.

中文翻译：

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水稻中Os8N3的CRISPR / Cas9靶向诱变，赋予了对米氏黄单胞菌PV的抗性。水稻

背景

基因组编辑工具对于功能基因组学研究和生物技术应用非常重要。最近，用于基因敲除的簇状规则间隔的短回文重复序列（CRISPR）/ CRISPR相关蛋白9（Cas9）系统已成为最有效的基因组编辑工具。以前有报道说，在水稻植株中，Os8N3基因的敲低导致对米生黄单胞菌pv的抗性增强。米粉（Xoo），同时显示出异常的花粉发育。

结果

为了增强对Xoo的抗性，采用了CRISPR / Cas9系统来敲除水稻Os8N3。对T ₀，T ₁，T ₂和T ₃转基因水稻植株的基因型和经编辑的Os8N3进行分析后发现，该突变已传递至后代，纯合突变体显示出对Xoo的抗性显着增强。通过在T _1中分离，证实了CRISPR / Cas9介导的Os8N3基因编辑的稳定传播，而没有转移的DNA（T-DNA）代。关于许多研究的农艺性状，包括花粉发育，在温室生长条件下，纯合突变体与非转基因对照植物之间没有显着差异。

结论

这项研究的数据表明，CRISPR / Cas9介导的Os8N3版本可以成功用于非转基因作物改良。