精确的序列插入或植物中的替代在技术上具有挑战性,但在农作物育种中至关重要,因为许多农艺性状都受DNA碎片变化的影响。尽管已连续优化了素数(PE)以改善练习剂(Jiang etal。,2022; li etal。,2022a,b; Zong et al。,2022),但对于靶向插入或更换长时间的靶向插入还是不明显的。Similar strategies, twinPE (Anzalone et al ., 2022 ) and GRAND editing (Wang et al ., 2022 ), in which a pairofPEguideRNAs(pegRNAs)arepartially complementarytoeach other in their reverse transcriptase template (RTT) but are not homologous to the genomic sequences, were recently developed to facilitate longer sequence insertion (Figure 1a ).HPPD抑制剂除草剂(例如B-三酮)有效地控制出现的抗性杂草。水稻中的HIS1基因赋予了对三酮除草剂的广谱耐药性,而在Triketone敏感的Indica品种中发现了功能失调的HIS1等位基因(Maeda等,2019)。A genetic survey for 631 Indica varieties commonly used in rice breeding revealed that the 28-bp deletion is widely distributed, including 50.7% 3-line restorers, 40.7% 2-line restorers and 18.1% conventional varieties (Lv et al ., 2021 ), which causes a huge risk for applying HPPD-inhibitor herbicides in Indica rice cultivating area.s1035是一种精英常规的Indica品种,由于HIS1的28 bp缺失,对Triketone敏感。PE和大编辑策略已被测试以插入28 bp