Transformation of CRISPR/Cas9 expression construct to enhance saturated fatty acid synthesis in oil palm for biofuel production

Abstract

New technology innovations and the utilization of elite varieties are necessary for oil palm intensification to improve productivity. In addition to productivity improvement, oil palm intensification is also directed to improve the quality of crude palm oil (CPO) to be used in biofuel production. The modification of the fatty acid biosynthesis pathway through genetic engineering is one of the most promising approaches. The research aims to apply a genetic transformation to oil palm calli and to improve success regeneration of the transformed explants. A CRISPR/Cas9 expression construct to edit the PTE (palmitoyl-acyl carrier protein thioesterase) gene, which protein is involved in the formation of palmitic acid, was generated in the previous research. In this study, the genetic transformation of the construct into 4month-old oil palm calli was conducted via Agrobacterium tumefaciens. The transformed calli were subsequently subcultured into selection media containing 100 ppm of cefotaxime and 50 ppm of kanamycin antibiotics. After several weeks, the transformed calli were transferred into DF10 media without antibiotics. Non-transformed calli were able to grow on the selection media with antibiotics but still in the globular phase. In contrast, the transformed calli on the selection media were able to grow to the scutellar phase with a compact structure in the embryogenic callus development stages. The next step is to optimize the transformation method as well as the regeneration media composition.