AUTHOR=Norouzi Moshen , Nazarain-Firouzabadi Farhad , Ismaili Ahmad , Ahmadvand Rahim , Poormazaheri Helen 

TITLE=CRISPR/Cas StNRL1 gene knockout increases resistance to late blight and susceptibility to early blight in potato

JOURNAL=Frontiers in Plant Science

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1278127

DOI=10.3389/fpls.2023.1278127

ISSN=1664-462X

ABSTRACT=<p>With the development of genome editing technologies, editing susceptible genes is a promising method to modify plants for resistance to stress. <italic>NPH3/RPT2-LIKE1</italic> protein (<italic>NRL1</italic>) interacts with effector Pi02860 of <italic>Phytophthora infestans</italic> and creates a protein complex, promoting the proteasome-mediated degradation of the guanine nucleotide exchange factor SWAP70. SWAP70, as a positive regulator, enhances cell death triggered by the perception of the <italic>P. infestans</italic> pathogen-associated molecular pattern (PAMP) INF1. Using a clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, a construct was made to introduce four guide RNAs into the potato cultivar Agria. A total of 60 putative transgenic lines were regenerated, in which 10 transgenic lines with deletions were selected and analyzed. A mutant line with a four-allelic knockdown of <italic>StNRL1</italic> gene was obtained, showing an ~90% reduction in <italic>StNRL1</italic> expression level, resulting in enhanced resistance to <italic>P. infestans</italic>. Surprisingly, mutant lines were susceptible to <italic>Alternaria alternata</italic>, suggesting that <italic>StNRL1</italic> may play a role as a resistance gene; hence, silencing <italic>StNRL1</italic> enhances resistance to <italic>P. infestans</italic>.</p>