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Enhancing resistance to bacterial blight in rice using CRISPR-based base editing technology
Friday, 2024/10/04 | 08:18:04
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Chenhao Li, Bo Liu, Hansong Dong, Bing Yang. The Crop Journal; Available online 21 September 2024 AbstractBacterial blight (BB), caused by Xanthomonas oryzae pathovar oryzae (Xoo), poses a significant threat to rice production, particularly in Asia and West Africa. Breeding resistance against BB in elite rice varieties is crucial to advancing rice breeding program and supporting smallholder farmers. Transcription Activator-Like effectors (TALes) are key virulence factors in Xoo, with some targeting the susceptibility (S) genes such as the sugar transporter SWEET genes in rice. Among these, SWEET14 is an important S gene, with its promoter bound by the TALe TalC which exists across all sequenced African Xoo isolates. In the present study, we utilized CRISPR/Cas9-based cytidine and adenine base editors to alter the effector binding element (EBE) of TalC in the promoter of SWEET14 in rice cultivars Kitaake, IR24, and Zhonghua 11. Mutations with C to T changes in EBE led to reduced SWEET14 induction by TalC-containing Xoo strains, resulting in resistance to African Xoo isolates reliant on TalC for virulence. Conversely, A to G changes retained SWEET14 inducibility and susceptibility to Xoo in edited lines. Importantly, no off-target mutations were detected at predicted sites, and the edited lines exhibited no obvious defects in major agronomic traits in Kitaake. These results underscore the effectiveness of base editing systems for both molecular biology research and crop improvement endeavors.
See https://www.sciencedirect.com/science/article/pii/S2214514124001831
Figure 1: The virulence of TalC depends on the induction of SWEET14 in rice. (A) The diagram of the SWEET14 gene structure. Black blocks indicate the exons, black lines between exons indicate the introns, and grey blocks indicate the untranslated regions. The numbers indicate the lengths of individual regions. The EBE sequence for TalC is shown. (B) The disease phenotypes of Kitaake are caused by ME2 and the TalC-containing strain ME2(TalC). Representative diseased leaves were photographed 14 d after inoculation, and red arrowheads indicate the edges of the lesions. (C) The gel images of semi-quantitative RT-PCR with SWEET14 after inoculation with ME2(TalC), with the rice actin gene used as an internal control.
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