Electrical charge-mediated transformation for efficient plant genome editing

Abstract

Genome editing technologies possess significant potential to enhance plant breeding; however, the delivery of editing constructs poses challenges in numerous crop species due to their resistance to transformation or tissue culture. This study introduces an innovative approach for the delivery of ribonucleoprotein (RNP) complexes and plasmid vectors into intact, regenerable faba bean plant tissues. This is accomplished by applying an electric current that makes plant cell walls and membranes permeable, thereby allowing the entry of macromolecular constructs into the cell and nucleus. This study assessed the efficacy of electric pulse-mediated transfection in faba bean by generating stable GFP-expressing faba bean plants. Furthermore, we incorporated it into faba bean leaf tissue and demonstrated its application in both embryos and leaf tissues. We demonstrate DNA-free genome editing by targeting the endogenous phytoene desaturase gene (PDS), achieving a mutation success rate of 50%. This method is efficient and economical, necessitating limited technical training. It is applicable to both leaf and embryo tissues, thereby enhancing its utility for crop improvement. This technique shows potential for the development of new crop varieties that can more effectively address global climate challenges.