The study demonstrates the successful application of the CRISPR/Cas9/sgRNA system for targeted gene modification in Arabidopsis, tobacco, rice, and sorghum. The Cas9/sgRNA system was delivered to plant cells using Agrobacterium tumefaciens, which introduced genes encoding Cas9, sgRNA, and a non-functional mutant green fluorescent protein (GFP). The Cas9/sgRNA complex cleaved the target DNA in the mutant GFP gene, leading to error-prone DNA repair by non-homologous end-joining (NHEJ), which restored the functional GFP gene. DNA sequencing confirmed the mutations at the target site. In rice, the Cas9/sgRNA system was used to target the promoter region of the bacterial blight susceptibility genes OsSWEET14 and OsSWEET11, resulting in mutated DNA sequences at the target sites. In sorghum, the system was used to target a non-functional DsRED2 gene, which was converted into a functional gene through Cas9/sgRNA-mediated cleavage and NHEJ repair. The study also showed that the Cas9/sgRNA system can be used for gene modification in plant cells without the need for Agrobacterium, as demonstrated by the successful transformation of rice protoplasts using PEG-mediated delivery. The results indicate that the Cas9/sgRNA system is functional in both model plant species (Arabidopsis and tobacco) and major crop species (rice and sorghum). The system enables targeted gene modification, including the restoration of functional genes, and has the potential to be a powerful tool for plant genetic engineering in scientific and agricultural applications. The study provides compelling evidence that the Cas9/sgRNA system is fully functional in plant cells and suggests its promise for future applications in plant biology and crop improvement.The study demonstrates the successful application of the CRISPR/Cas9/sgRNA system for targeted gene modification in Arabidopsis, tobacco, rice, and sorghum. The Cas9/sgRNA system was delivered to plant cells using Agrobacterium tumefaciens, which introduced genes encoding Cas9, sgRNA, and a non-functional mutant green fluorescent protein (GFP). The Cas9/sgRNA complex cleaved the target DNA in the mutant GFP gene, leading to error-prone DNA repair by non-homologous end-joining (NHEJ), which restored the functional GFP gene. DNA sequencing confirmed the mutations at the target site. In rice, the Cas9/sgRNA system was used to target the promoter region of the bacterial blight susceptibility genes OsSWEET14 and OsSWEET11, resulting in mutated DNA sequences at the target sites. In sorghum, the system was used to target a non-functional DsRED2 gene, which was converted into a functional gene through Cas9/sgRNA-mediated cleavage and NHEJ repair. The study also showed that the Cas9/sgRNA system can be used for gene modification in plant cells without the need for Agrobacterium, as demonstrated by the successful transformation of rice protoplasts using PEG-mediated delivery. The results indicate that the Cas9/sgRNA system is functional in both model plant species (Arabidopsis and tobacco) and major crop species (rice and sorghum). The system enables targeted gene modification, including the restoration of functional genes, and has the potential to be a powerful tool for plant genetic engineering in scientific and agricultural applications. The study provides compelling evidence that the Cas9/sgRNA system is fully functional in plant cells and suggests its promise for future applications in plant biology and crop improvement.