Functional inhibition of the StERF3 gene by dual targeting through CRISPR/Cas9 enhances resistance to the late blight disease in Solanum tuberosum L

• Published in: Molecular biology reports Vol. 49; no. 12; pp. 11675 - 11684
• Main Authors: Razzaq, Hafiza Arooj, Ijaz, Siddra, Haq, Imran Ul, Khan, Iqrar Ahmad
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-12-2022; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Biomedical and Life Sciences; CRISPR; Cultivars; Disease resistance; Fungicides; Genes; Genome editing; Genomes; Histology; Late blight; Life Sciences; Morphology; Mutants; Original Article; Plant diseases; Potatoes; Sequence analysis; CRISPR; StERF3-loss-of-function mutants; Indel detection by amplicon analysis; Detached leaf assay; Cas9
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-022-07958-1

Background
Disease-resistant cultivars are the best solution to get their maximum yield potential and avoid fungicide application. There is no doubt about the contribution, and use of
R
genes (resistance genes) in resistance development in plants, while
S
genes (susceptibility genes) also hold a strong position in pathogenesis by resistance repression, and their loss of function contributes to enhanced resistance. Hence, we attempted to knock out the function of
the StERF3
gene in potatoes through CRISPR/Cas9-based genome editing and investigated the CRISPR/Cas9 approach as strategic control against late blight disease in potato plants.
Methods and results
The
StERF3
gene was edited in late blight susceptible cv. Lady Rosetta. Full allelic edited plants were identified through DnpI, and N1aIV mediated restriction digestion and then further analyzed through Indel Detection by Amplicon Analysis. Sequence analysis of targeted plants for indel identification showed full allelic editing. The detached leaf assay of full allelic edited plants demonstrated the role of the
StERF3
gene in susceptibility to late blight in potatoes. In planta disease assay also showed reduced, slowed, and delayed disease progression in StERF3-loss-of-function mutants compared to wild-type (control) plants. Less fungal biomass was quantified in knockouts through Real-time qPCR that supported less susceptibility of edited plants to late blight. Besides, relatively high expression of pathogens-related genes, StPR1, and StNPR1, were also observed in StERF3-loss-of-function mutants compared to the corresponding control.
Conclusion
The results showed the functional inhibition of
StERF3
genes using the CRISPR/Cas9 approach. The functional knockouts (
StERF3
gene-edited potato plants) revealed enhanced resistance against
Phytophthora infestans
, thereby demonstrating the best strategic control for late blight disease in potato plants.