OsPUB9 Gene Edited by CRISPR/Cas9 Enhanced Resistance to Bacterial Leaf Blight in Rice ( Oryza sativa L.)

• Published in: International journal of molecular sciences Vol. 25; no. 13; p. 7145
• Main Authors: Kim, Me-Sun, Le, Van Trang, Jung, Yu Jin, Kang, Kwon-Kyoo, Cho, Yong-Gu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.07.2024
• Subjects: bacterial leaf blight; Carbenicillin; Cell growth; CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Disease Resistance - genetics; E3 ligase; Gene Editing - methods; Genes; Genome editing; Genomes; Kinases; Mutation; Oryza - genetics; Oryza - microbiology; OsPUB; Pathogens; Plant Diseases - genetics; Plant Diseases - microbiology; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified - genetics; Proteins; Proteolysis; R&D; Research & development; Ubiquitin; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - metabolism; ubiquitination; South Korea; E3 ligase; CRISPR/Cas9; ubiquitination; bacterial leaf blight; OsPUB
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25137145

Ubiquitination plays a crucial role in regulating signal pathways during the post-translation stage of protein synthesis in response to various environmental stresses. E3 ubiquitin ligase has been discovered to ultimately control various intracellular activities by imparting specificity to proteins to be degraded. This study was conducted to confirm biological and genetic functions of the U-box type E3 ubiquitin ligase (PUB) gene against biotic stress in rice ( L.). gene-specific sgRNA were designed and transformants were developed through -mediated transformation. Deep sequencing using callus was performed to confirm the mutation type of T plants, and a total of three steps were performed to select null individuals without T-DNA insertion. In the case of the gene-edited line, a one bp insertion was generated by gene editing, and it was confirmed that early stop codon and multiple open reading frame (ORF) sites were created by inserting thymine. It is presumed that ubiquitination function also changed according to the change in protein structure of U-box E3 ubiquitin ligase. The gene-edited null lines were inoculated with bacterial leaf blight, and finally confirmed to have a resistance phenotype similar to Jinbaek, a bacterial blight-resistant cultivar. Therefore, it is assumed that the amino acid sequence derived from the gene is greatly changed, resulting in a loss of the original protein functions related to biological mechanisms. Comprehensively, it was confirmed that resistance to bacterial leaf blight stress was enhanced when a mutation occurred at a specific site of the gene.