| Autor: |
Phad, Archana P., Takate, Umesh B., Rawal, Shuban K., Pyati, Prashant S., Lomate, Purushottam R. |
| Zdroj: |
Journal of Crop Science & Biotechnology; Mar2024, Vol. 27 Issue 2, p249-259, 11p |
| Abstrakt: |
Genome editing using CRISPR/Cas9 has become powerful tool for enabling targeted genetic changes into crop species. The present study was aimed to establish genome editing in eggplant (Solanum melongena) via generating phytoene desaturase (SmPDS) knockout plants using CRISPR/Cas9 technology. A binary vector containing SmPDS-specific gRNA was designed and transformed into eggplant inbred line using Agrobacterium-mediated transformation. The efficiency of Agrobacterium-mediated transformation was found to be 71%. Three types of regenerated plants (albino, chimeric, and non-edited green plants) were observed that growing on kanamycin. The edited plants exhibited a characteristic albino phenotype, which confirmed the efficient knockout of the PDS gene. The knockout frequency of CRISPR/Cas9 was found to be 67% using the binary construct SmPDSC1. The PCR amplification confirmed the integration of CRISPR/Cas9 T-DNA into the genome of transformed plants. The sequence analysis revealed a 15-nucleotide deletion in knockout target region in all transgenic events that displayed albino phenotype. The T7 endonuclease assay validated CRISPR/Cas9-induced mutations in the SmPDS gene. A three-primer system was used to determine the homozygosity of mutant plants. Results showed that all the albino plants tested were homozygous. Our study demonstrates the successful use of the CRISPR/Cas9 system to generate PDS knockout mutants in eggplant, which has potential applications in plant biotechnology. This work will be beneficial for functional genomics research in eggplant, and the CRISPR/Cas9 system can be used for generating desirable traits in eggplant. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
|
Full text from SpringerLink