Using genetic resistance against bacterial blight (BB) caused by Xanthomonas oryzae pathovar oryzae (Xoo) is a major objective in rice breeding programmes. Prime editing (PE) has the potential to create novel germplasm against Xoo. Here, we use an improved prime-editing system to implement two new strategies for BB resistance. Knock-in of TAL effector binding elements (EBE) derived from the BB susceptible gene SWEET14 into the promoter of a dysfunctional executor R gene xa23 reaches 47.2% with desired edits including biallelic editing at 18% in T₀ generation that enables an inducible TALE-dependent BB resistance. Editing the transcription factor TFIIA gene TFIIAγ5 required for TAL effector-dependent BB susceptibility recapitulates the resistance of xa5 at an editing efficiency of 88.5% with biallelic editing rate of 30% in T₀ generation. The engineered loci provided resistance against multiple Xoo strains in T₁ generation. Whole-genome sequencing detected no OsMLH1dn-associated random mutations and no off-target editing demonstrating high specificity of this PE system. This is the first-ever report to use PE system to engineer resistance against biotic stress and to demonstrate knock-in of 30-nucleotides cis-regulatory element at high efficiency. The new strategies hold promises to fend rice off the evolving Xoo strains and protect it from epidemics.

中文翻译：

---

高效引物编辑实现了广谱抗水稻白叶枯病的新策略

利用遗传抗性对抗由米黄单胞菌（ Xanthomonas oryzae pathovar oryzae）（Xoo）引起的白叶枯病（BB ）是水稻育种计划的一个主要目标。Prime 编辑 (PE) 有潜力创造针对Xoo 的新种质。在这里，我们使用改进的素数编辑系统来实施两种新的 BB 抵抗策略。将源自 BB 易感基因SWEET14的 TAL 效应结合元件 (EBE) 敲入功能失调的执行器R基因xa23的启动子中，达到 47.2%，并实现所需的编辑，包括 T ₀中 18% 的双等位基因编辑产生可诱导的依赖于 TALE 的 BB 抗性。编辑TAL效应子依赖性BB易感性所需的转录因子TFIIA基因TFIIAγ5概括了xa5的抗性，在T ₀代中编辑效率为88.5％，双等位基因编辑率为30％。工程基因座在 T _{1代中提供了针对多种}Xoo菌株的抗性。全基因组测序未检测到OsMLH1dn-相关的随机突变和无脱靶编辑，证明了该 PE 系统的高度特异性。这是第一份利用 PE 系统设计生物胁迫抗性并证明高效敲入 30 个核苷酸顺式调控元件的报告。新策略有望抵御不断进化的Xoo菌株，并保护其免受流行病的侵害。