Agrobacterium tumefaciens-Mediated Plant Transformation and Gene Editing in Rice

Bottlenecks in plant transformation and regeneration have slowed progress in applying CRISPR/Cas9-based genome editing for crop improvement. Rice (Oryza sativa L.) has highly efficient temperate japonica transformation protocols, along with reasonably efficient indica protocols using immature embryos. However, rapid and efficient protocols are not available for transformation and regeneration in tropical japonica varieties, even though they represent most of the rice production in the USA and South America, along with some regions in Asia. This chapter describes a protocol for CRISPR/Cas9 gene editing using Agrobacterium-mediated transformation for the tropical japonica rice cultivar Presidio leading to knock-out mutations in the phytoene desaturase (PDS) gene.