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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 T 0 , T 1 , T 2 , and T 3  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 T 1  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.    Electronic supplementary material  The online version of this article (10.1186/s12284-019-0325-7) contains supplementary material, which is available to authorized users.

CRISPR/Cas9-targeted mutagenesis of Os8N3 in rice to confer resistance to Xanthomonas oryzae pv. oryzae