Generating herbicide resistant and dwarf rice germplasms through precise sequence insertion or replacement
Xiaoxia Liu1,2†, Yingying Wang3,4†, Hongzhi Wang3,4†, Yuxin He3,4, Yijiao Song5, Zuren Li6, Ming Li1, Chuang Wei1, Yinghui Dong1, Li Xue1, Jinshan Zhang7, Jian-Kang Zhu1,8* and Mugui Wang3*
Plant Biotechnology Journal；2023；IF 13.8
DOI: 10.1111/pbi.14225.
Abstract
Many agronomic traits are affected by DNA fragment variations, such as deletion, insertion, duplication，and replacements. These variations are difficult to correct by base editors and traditional prime editors. The twinPE and GRAND editing strategies were developed recently for insertion or replacement of larger DNA fragments. In this study, we tested GRAND editing strategy for targeted insertion and replacement of DNA fragments in rice genome. We successfully inserted a 28-bp fragment for repairing a dysfunctional his1 allele in triketone-sensitive Indica varieties to restore their herbicide resistance. We also introduced the glyphosate-resistant TIPS (T173I+P177S) mutations into the OsEPSPS gene, and replaced a 28-bp fragment in the OsSLR1 gene to create dwarf germplasm at efficiencies up to 50%. Our results show that GRAND editing is an effective strategy for insertion or replacement of DNA fragments to create herbicide resistant and other germplasms in rice.
