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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Research » Research » Publications at this Location » Publication #398984
Research Project: Improvement of Biotic Stress Resistance in Durum and Hard Red Spring Wheat Using Genetics and Genomics

Location: Cereal Crops Research

Title: Targeting wheat disease susceptibility genes in wheat through wide hybridization with maize expressing Cas9 and guide RNA

Author
item KARMACHARYA, ANIL - North Dakota State University
item Li, Dandan
item LENG, YUEQIANG - North Dakota State University
item SHI, GONGJUN - North Dakota State University
item LIU, ZHAOHUI - North Dakota State University
item Yang, Shengming
item DU, YANG - Valley City State University
item DAI, WENHAO - North Dakota State University
item ZHONG, SHAOBIN - North Dakota State University

Submitted to: Molecular Plant-Microbe Interactions
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/2/2023
Publication Date: 4/4/2023
Citation: Karmacharya, A., Li, D., Leng, Y., Shi, G., Liu, Z., Yang, S., Du, Y., Dai, W., Zhong, S. 2023. Targeting wheat disease susceptibility genes in wheat through wide hybridization with maize expressing Cas9 and guide RNA. Molecular Plant-Microbe Interactions. https://doi.org/10.1094/MPMI-01-23-0004-SC.
DOI: https://doi.org/10.1094/MPMI-01-23-0004-SC

Interpretive Summary: Elimination of susceptibility genes is an effective means to improve disease resistance in wheat. Although the current gene-editing technology is a powerful tool for gene mutation, this technology can only be applied to a very limited number of wheat varieties due to technical issues. In the present study, we transferred the gene-editing elements targeting two wheat susceptibility genes to maize. The decedents of transgenic maize plants were used to pollinate a wheat variety which is not compatible with the traditional gene-editing technology. Sequence analysis indicated that we successfully obtained mutations in both susceptibility genes with high efficiencies. Therefore, our study provides geneticists an upgraded protocol to broaden the gene-editing spectrum in wheat, as well as resistant materials for breeders to improve disease resistance.

Technical Abstract: Two wheat genes (TaHRC and Tsn1) were targeted using wide hybridization between wheat and maize expressing Cas9 and guide RNA. For each gene, two target sites were selected and used to make a gene construct that contains CRISPR/Cas9-mediated genome editing machinery. The gene constructs were used to transform the hybrid maize Hi-II to generate T0 and T1 plants, which were used to pollinate emasculated wheat spikes. Embryos were rescued in vitro to generate haploid plants. PCR amplification and sequencing indicated that 15-33% of the haploid plants contained the target gene with mutations at the target sites. This approach should be very useful for targeting any other susceptibility genes to improve wheat disease resistance without regulatory issues.