Knockout Mutants of OsPUB7 Generated Using CRISPR/Cas9 Revealed Abiotic Stress Tolerance in Rice

• Published in: International journal of molecular sciences Vol. 24; no. 6; p. 5338
• Main Authors: Kim, Me-Sun, Ko, Seo-Rin, Jung, Yu Jin, Kang, Kwon-Kyoo, Lee, Yung-Jo, Cho, Yong-Gu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 10-03-2023; MDPI
• Subjects: CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Drought resistance; Droughts; E3 ligase; Flowers & plants; Gene expression; Gene Expression Regulation, Plant; Genetic modification; Kinases; Mutation; Oryza - metabolism; Oryza sativa; OsPUB; Phenotypes; Plant Breeding; Plant Proteins - metabolism; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Protein turnover; Proteins; Rice; Salinity; Salinity effects; Seeds; Stress, Physiological - genetics; T-DNA; transcriptomics; Ubiquitin; Ubiquitin-protein ligase; Ubiquitination; transcriptomics; E3 ligase; CRISPR/Cas9; ubiquitination; OsPUB
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24065338

Plants produce and accumulate stress-resistant substances when exposed to abiotic stress, which involves a protein conversion mechanism that breaks down stress-damaged proteins and supplies usable amino acids. Eukaryotic protein turnover is mostly driven by the ubiquitination pathway. Among the three enzymes required for protein degradation, E3 ubiquitin ligase plays a pivotal role in most cells, as it determines the specificity of ubiquitination and selects target proteins for degradation. In this study, to investigate the function of
(Plant U-box gene in
), we constructed a CRISPR/Cas9 vector, generated
gene-edited individuals, and evaluated resistance to abiotic stress using gene-edited lines. A stress-tolerant phenotype was observed as a result of drought and salinity stress treatment in the T
gene-edited null lines (PUB7-GE) lacking the T-DNA. In addition, although PUB7-GE did not show any significant change in mRNA expression analysis, it showed lower ion leakage and higher proline content than the wild type (WT). Protein-protein interaction analysis revealed that the expression of the genes (
,
,
, and
) known to be involved in stress increased in PUB7-GE and this, by forming a 1-node network with
and
, acted as a negative regulator of drought and salinity stress. This result provides evidence that
will be a useful target for both breeding and future research on drought tolerance/abiotic stress in rice.