CRISPR/Cas9-mediated genome editing in diploid and tetraploid potatoes

CRISPR/Cas9 is a unique technology that has enabled researchers to edit genomes. The current study was conducted to investigate the efficiency of genome editing in diploid and tetraploid potato varieties. The stem internodes of AGB and M6 plants (5-6 weeks old) were infected with Agrobacterium strain LBA4404, harboring CRISPR/Cas9 vector, expressing a particular guide RNA against vacuolar invertase; and Cas9 endonuclease. Both tetraploid and diploid potatoes showed encouraging regenerative response on selection media. The transformation efficiency of 11.7% was obtained in potatoes in this study, whereas an indel% was found to be 6 for diploid and 5 for tetraploid variety via Sanger sequencing data. The regenerated plantlets were shifted to culture tubes where M6 diploid variety needed indole-3-butyric acid (IBA) for root development in comparison to tetraploid variety that rooted without IBA. All molecular analysis confirmed the integration of cassette and expression of Cas9 in primary transformants. Overall, the results concluded that the adopted transformation protocol regenerated plants with edited targeted genes that can be further used in an efficient potato breeding programme.