Enhanced Rice Blast Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922

By Fujun Wang, Chunlian Wang, Pi-qing Liu, C. Lei, Wei-wei Hao, Ying Gao, Yaoguang Liu, Kaijun Zhao. PLoS ONE, 2016

667 citations recorded by the source corpus.

Rice blast is one of the most destructive diseases affecting rice worldwide. The adoption of host resistance has proven to be the most economical and effective approach to control rice blast. In recent years, sequence-specific nucleases (SSNs) have been demonstrated to be powerful tools for the improvement of crops via gene-specific genome editing, and CRISPR/Cas9 is thought to be the most effective SSN. Here, we report the improvement of rice blast resistance by engineering a CRISPR/Cas9 SSN (C-ERF922) targeting the OsERF922 gene in rice. Twenty-one C-ERF922-induced mutant plants (42.0%) were identified from 50 T0 transgenic plants. Sanger sequencing revealed that these plants harbored various insertion or deletion (InDel) mutations at the target site. We showed that all of the C-ERF922-induced allele mutations were transmitted to subsequent generations. Mutant plants harboring the desired gene modification but not containing the transferred DNA were obtained by segregation in the T1 and T2 generations. Six T2 homozygous mutant lines were further examined for a blast resistance phenotype and agronomic traits, such as plant height, flag leaf length and width, number of productive panicles, panicle length, number of grains per panicle, seed setting percentage and thousand seed weight. The results revealed that the number of blast lesions formed following pathogen infection was significantly decreased in all 6 mutant lines compared with wild-type plants at both the seedling and tillering stages. Furthermore, there were no significant differences between any of the 6 T2 mutant lines and the wild-type plants with regard to the agronomic traits tested. We also simultaneously targeted multiple sites within OsERF922 by using Cas9/Multi-target-sgRNAs (C-ERF922S1S2 and C-ERF922S1S2S3) to obtain plants harboring mutations at two or three sites. Our results indicate that gene modification via CRISPR/Cas9 is a useful approach for enhancing blast resistance in rice.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/730bf934ea734cdc5780074a1d2af0f30facb559

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Citations in the corpus

Improvement of salinity stress tolerance in rice: Challenges and opportunities (2016, methodology): “Recent studies have shown improved resistance to rice blast by mutating the OsERF922 gene using the CRISPR/Cas system [187].”
Genome engineering for crop improvement and future agriculture. (2021, positive): “For example, CRISPR-Cas9 was successfully used to knock out OsERF922 , a negative regulator of fungal blast resistance in plants, leading to the production of blast-resistant rice (Wang et al., 2016).”
Genome Editing—Principles and Applications for Functional Genomics Research and Crop Improvement (2017, background): “Knockout of the ERF transcription factor OsERF922, a negative regulator of rice blast resistance, resulted in enhanced resistance (Wang et al., 2016a).”
Past and Future Use of Wild Relatives in Crop Breeding (2017, background): “Although these methods are still in their infancy, their increased precision will likely result in substantial efficiency gains in crop improvement processes (Hu et al., 2016; Wang et al., 2016).”
CRISPR/Cas Genome Editing and Precision Plant Breeding in Agriculture. (2019, background): “Rice blast is a destructive fungal disease; blast-resistant rice was obtained via knockout of OsERF922, an ethyleneresponsive factor transcription factor gene (164).”
The Pivotal Role of Ethylene in Plant Growth (2018, background): “Magnaporthe oryzae, but no difference with the wild type for several yield-related traits [51].”