Konstruksi Dan Mutasi Terarah Gen Pengkode Enzim Pensintesa Etilen ACO-1 Menggunakan Crispr/Cas9 Untuk Meningkatkan Usia Simpan Buah Tanaman Tomat

Abstract

Ethylene is a hormone that plays a role in the ripening process of climacteric fruits, particularly tomatoes. The ACO1 gene is part of the ACO (ACC Oxidase) gene family, which plays a role in the final stage of ethylene biosynthesis in plants; ACC (1-aminocyclopropane-1-carboxylic acid) is converted into ethylene (C2H4) by the ACO gene. However, high ethylene accumulation can accelerate the ripening rate of tomato fruit, resulting in a relatively shorter shelf life. This study employed genome editing of the ACO1 gene using the pRGEB32 plasmid mediated by Agrobacterium tumefaciens strain GV3101 to achieve ACO1 knockout. This study aimed to determine the effect of ACO1 gene knockout using the Agrobacterium tumefaciens-mediated pRGEB32 plasmid on ethylene production in Biromaru tomato plants through genetic transformation. This study was conducted at the Agrobiotechnology Laboratory, Faculty of Agriculture, University of Jember. The results were analyzed using simple descriptive statistics to assess the transformation efficiency of tomato plants with the ACO1 target gene inserted via pRGEB32.sgRNAI and pRGEB32.sgRNAII which ranged from 0.01% to 0.02%. The study resulted in the development of sgRNA designs and plasmid constructs.