Received: 29-11-2021
Accepted: 05-04-2022
DOI:
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Genetic Identification of Target Mutationsin OsSWEET14 Promoter-edited TBR225 Rice Lines
,
Pham Thu Hang
2
,
Cao Le Quyen
2
,
Pham Thi Van
2
,
Pham Xuan Hoi
2
,
Nguyen Duy Phuong
2
Keywords
Bacterial leaf blight disease, CRISPR/Cas9, OsSWEET14, TBR225, Xanthomonas oryzae
Abstract
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 ofrice cultivars. In a previous study, we generated some transgenic TBR225 rice lines containing a single copy of CRISPR/Cas9 expression construct for editing AvrXa7/Tal5EBE on the promoter OsSWEET14. In this study, we continued to evaluate genotype and phenotype of T1edited TBR225 lines to determinethe genetic stability and phenotypic effective of the CRISPR/Cas9-induced OsSWEET14mutations. The genotypic analysis showed that 65%of transgenic TBR225 plants harbored the targeted OsSWEET14mutation, including nucleotide (Nu) insertion and deletion. The OsSWEET14mutations 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 OsSWEET14promoter did not affect the growth and yield of gene-edited rice plants. Some T1TBR225 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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