Creation of gene-speciifc rice mutants by AvrXa23-based TALENs

摘要

Transcription activator-like effector (TALE) nucleases (TALENs) are increasingly used as a powerful tool for genome edit-ing in a variety of organisms. We have previously cloned the TALE-coding geneavrXa23 fromXanthomonas oryzaepv. oryzae and developed an AvrXa23-based assembly system for designer TALEs or TALENs. Here, we exploit TALENs to induce mutagenesis of the rice ethylene response factor (ERF) transcription factorOsERF922 for testing the gene-editing efifciency of AvrXa23-based TALENs system. A pair of TALENs (T-KJ9/KJ10) was assembled and their nuclease activities were ifrst conifrmed in rice protoplast transient assay. The TALENs-expressing construct pT-KJ9/KJ10 was then used for rice transformation. We observed targeting somatic mutagenesis frequency of 15.0% in positive transgenic rice cali and obtained two mutant plants with nucleotide deletion or insertion at the designer target region. Our work demonstrates that the AvrXa23-based TALENs system can be used for site-speciifc genome editing in rice.