Understanding host-pathogen interactions is key in breeding disease-resistant crops, including bacterial leaf blight. As a key virulence strategy to cause bacterial leaf blight, Xanthomonas oryzae pv. oryzae (Xoo) secretes transcription activator-like effectors (TALEs) to control the expression of host target genes through binding to the effector-binding elements (EBEs) located on the promoter regions. Precise mutagenesis at the TALE-interaction sites on the host genes by using gene editing tools such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRSIPR/Cas9) has been seen as a promising way to improve BLB resistance of rice cultivars. In a previous study, we generated some transgenic TBR225 rice lines containing a single copy of CRISPR/Cas9 expression construct for editing AvrXa7/Tal5 EBE on the promoter OsSWEET14. In this study, we continued to evaluate genotype and phenotype of T1 edited TBR225 lines to determine the genetic stability and phenotypic effective of the CRISPR/Cas9-induced OsSWEET14 mutations. The genotypic analysis showed that 65%of transgenic TBR225 plants harbored the targeted OsSWEET14 mutation, including nucleotide (Nu) insertion and deletion. The OsSWEET14 mutations were stably inherited to the next generation with an expected segregation ratio consistent with Mendelian segregation (1:2:1). The preliminary phenotypic analysis suggested that the mutation of OsSWEET14 promoter did not affect the growth and yield of gene-edited rice plants. Some T1 TBR225 lines showed the resistance to Xoo isolate VXO_11 in the leaf-clipping experiment. The obtained results are a premise for development of the BLB-resistant TBR225 rice variety in the future.
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