During the dry season, declining soil moisture frequently limits water availability for potatoes (Solanum tuberosum L.), exposing plants to drought stress that disrupts normal growth and ultimately reduces tuber yield. Therefore, drought stress-tolerant potato cultivars are essential to address this alarming issue. Stomatal conductance plays a central role in plant responses to drought because it regulates both water losses through transpiration and CO₂ uptake for photosynthesis. Hence, optimizing stomatal conductance represents an effective strategy to improve water-use efficiency while sustaining plant growth under water-limited conditions. The Pain-1 gene with reduced expression promotes the degradation of sucrose into glucose and fructose through genome editing using CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9). This study aimed to identify the potato explant type with the highest transformation efficiency to support efficient CRISPR/Cas9-mediated editing of the Pain-1 gene. As a preliminary study, transformation efficiency analysis ensued through leaf and internode explants of potato clone BR1 plantlets used for genome editing. The transformation took place using the Agrobacterium tumefaciens strain EHA101 containing the pGW903 vector plasmid, target sequence, and two sgRNA designs (VIST2 and VIST3). Transformant status received assessment by identifying the presence of the Cas9 and hpt genes. The results showed internode explants exhibited significantly higher transformation efficiency (33.46%) than leaf explants (7.95%).
Potato (S. tuberosum L.), drought tolerance, genome editing, CRISPR/Cas9, explant, sgRNA, tuber yield
In potatoes (S. tuberosum L.), the efficient genome editing was highly dependent on the explant type. This preliminary study revealed a significant difference between the transformation efficiency of potato leaves and internodes. Internode explants consistently showed higher transformation efficiency across two different sgRNA designs. The results suggested the use of internodes as explants for genome editing in potatoes.