Transformation of the p53 plasmid carrying Cas9 to Chili Callus (Capsicum annum) via Agrobacterium tumefaciens

Abstract

Geminivirus is a virus that causes curly yellow disease in chili plants, which causes yield losses ranging from 20-100%. CRISPR/Cas9 facilitates the development of virus-resistant plants using genetic engineering technology. In this study, the process of transferring the plasmid p53/Cas9 to Agrobacterium was executed using the freeze-thawing technique. To validate the achievement of the transformation, an amplification process utilizing specific primers and an in-out primer combination was performed. We have achieved a transformation efficiency of 60%, as indicated by the PCR detection analysis of the putative recombinant callus. This outcome serves as a crucial starting point for the subsequent stages involved in transforming the p53 plasmid containing the Cas9 gene into chili (Capsicum annum) via Agrobacterium tumefaciens.