CRISPR/Cas9-mediated mutagenesis of VvMLO3 results in enhanced resistance to powdery mildew in grapevine (Vitis vinifera).

Printer-friendly versionPrinter-friendly versionPDF versionPDF version
TitleCRISPR/Cas9-mediated mutagenesis of VvMLO3 results in enhanced resistance to powdery mildew in grapevine (Vitis vinifera).
Publication TypeJournal Article
Year of Publication2020
AuthorsWan, D-Y, Guo, Y, Cheng, Y, Hu, Y, Xiao, S, Wang, Y, Wen, Y-Q
JournalHortic Res
Volume7
Issue1
Pagination116
Date Published2020 Aug 01
ISSN2662-6810
Abstract

Grapevine (Vitis vinifera), one of the most economically important fruit crops in the world, suffers significant yield losses from powdery mildew, a major fungal disease caused by Erysiphe necator. In addition to suppressing host immunity, phytopathogens modulate host proteins termed susceptibility (S) factors to promote their proliferation in plants. In this study, CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated 9) technology was used to enable the targeted mutagenesis of MLO (mildew resistance Locus O) family genes that are thought to serve as S factors for powdery mildew fungi. Small deletions or insertions were induced in one or both alleles of two grapevine MLO genes, VvMLO3 and VvMLO4, in the transgenic plantlets of the powdery mildew-susceptible cultivar Thompson Seedless. The editing efficiency achieved with different CRISPR/Cas9 constructs varied from 0 to 38.5%. Among the 20 VvMLO3/4-edited lines obtained, one was homozygous for a single mutation, three harbored biallelic mutations, seven were heterozygous for the mutations, and nine were chimeric, as indicated by the presence of more than two mutated alleles in each line. Six of the 20 VvMLO3/4-edited grapevine lines showed normal growth, while the remaining lines exhibited senescence-like chlorosis and necrosis. Importantly, four VvMLO3-edited lines showed enhanced resistance to powdery mildew, which was associated with host cell death, cell wall apposition (CWA) and HO accumulation. Taken together, our results demonstrate that CRISPR/Cas9 genome-editing technology can be successfully used to induce targeted mutations in genes of interest to improve traits of economic importance, such as disease resistance in grapevines.

DOI10.1038/s41438-020-0339-8
Alternate JournalHortic Res
PubMed ID34333528
Grant List31772264 / / National Natural Science Foundation of China (National Science Foundation of China) /
31772264 / / National Natural Science Foundation of China (National Science Foundation of China) /
31772264 / / National Natural Science Foundation of China (National Science Foundation of China) /
31772264 / / National Natural Science Foundation of China (National Science Foundation of China) /
31772264 / / National Natural Science Foundation of China (National Science Foundation of China) /
201810712097 / / Northwest A and F University (Northwest A & F University) /
201810712097 / / Northwest A and F University (Northwest A & F University) /
201810712097 / / Northwest A and F University (Northwest A & F University) /
201810712097 / / Northwest A and F University (Northwest A & F University) /
201810712097 / / Northwest A and F University (Northwest A & F University) /