Abstract
The frequency of climate change is increasing globally, which makes predictions challenging. Cold spells during the rice seedling stage can significantly reduce yield, prompting a constant need for cold-tolerant cultivars, which is a major breeding goal. However, the traditional crossbreeding of rice cultivars requires substantial time and effort. Recently, the application of CRISPR/Cas9 to reduce defects in elite cultivars has become a more cost-effective and time-efficient method for breeding cultivars than cross-breeding methods and can alleviate food insecurity. In the present study, CRISPR/Cas9-mediated genome editing was performed for OsCS511 a gene involved in cold susceptibility, identified using quantitative trait loci (QTL) mapping in Ilmi (Oryza sativa L. spp. Japonica cv. Ilmi). In Ilmi, CRISPR/Cas9 tool-edited OsCS511 homozygous lines were used in T0 and advanced generations in the field. CRISPR/Cas9 induced variations in the DNA sequence and plants with insertions or deletions compared to OsCS511 of Ilmi were selected as genome-edited lines. Agricultural traits, reactive oxygen species scavenging capacity, and stress-tolerance-related gene expression levels were evaluated under normal and cold stress conditions. Under normal conditions, all traits evaluated in the Ilmi and OsCS511 genome-edited lines exhibited similar results; however, when subjected to cold stress, the cold tolerance of OsCS511 genome-edited lines improved or reached the same level as that of Ilmi. OsCS511 genome-edited lines recovered and survived. From a breeding perspective, we suggest that CRISPR/Cas9 technology can precisely reduce defects in existing superior rice cultivars with high efficiency and speed.
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