Development of efficient methods for protoplast-to-plant regeneration in Coffea arabica and successful CRISPR-Cas9 editing in protoplasts
An efficient protocol for protoplast isolation from embryogenic calli and protoplast-to-plant regeneration in the allotetraploid species Coffea arabica is presented and polyethylene glycol-mediated transfection of coffee protoplasts with a CRISPR–Cas9 plasmid targeting the coffee xanthosine methyltransferase gene involved in caffeine biosynthesis is demonstrated.