CRISPR/Cas9-mediated Inhibition of Vacuolar Invertase (VInv) Gene in Potato

Abstract

Potato is the third most important food crop worldwide after the wheat and rice. It has potential to overcome the issues of world’s food security. Potato crop faces some challenges during storage as Cold Induce Sweetening (CIS). During cold storage, the accumulation of reducing sugars such as fructose and glucose takes place in potato tubers. These reducing sugars later react with free amino acid and produce dark-brown pigmentation on potato products during frying. That leads towards reduction of potato quality and consumption. The increased quantity of sucrose is further hydrolyzed by Vacuolar Invertase (VInv) gene into reducing sugars. VInv gene acts as a key player in inducing CIS in potato. The objective of study was cloning of CRISPR/Cas9 construct and successful inhibition of VInv gene function in potato. Agrobacterium-mediated transformation of VInv gene in potato tissues was performed. The knockout of VInv gene in transformed plants was analyzed by using specific primers through polymerase chain reaction (PCR). Real time polymerase chain reaction (RT-PCR) was carried out to analyze the expression of transgene in transgenic potato plants. The knockout of VInv gene using CRISPR/Cas9 strategy could maintain the quality of potato tubers during cold storage.