Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial blight of rice, employs the transcription activator-like effectors (TALEs) to induce the expression of the OsSWEET family of putative sugar transporter genes, which function in conferring disease susceptibility (S) in rice plants. To engineer broad-spectrum bacterial blight resistance, we used CRISPR/Cas9-mediated gene editing to disrupt the TALE-binding elements (EBEs) of two S genes, OsSWEET11 and OsSWEET14, in rice cv. Kitaake, which harbors the recessive resistance allele of Xa25/OsSWEET13. The engineered rice line MS14K exhibited broad-spectrum resistance to most Xoo strains with a few exceptions, suggesting that the compatible strains may contain new TALEs. We identified two PthXo2-like TALEs, Tal5_LN18 and Tal7_PXO61, as major virulence factors in the compatible Xoo strains LN18 and PXO61, respectively, and found that Xoo encodes at least five types of PthXo2-like effectors. Given that PthXo2/PthXo2.1 target OsSWEET13 for transcriptional activation, the genomes of 3000 rice varieties were analyzed for EBE variationsin the OsSWEET13 promoter, and 10 Xa25-like haplotypes were identified. We found that Tal5_LN18 and Tal7_PXO61 bind slightly different EBE sequences in the OsSWEET13 promoter to activate its expression. CRISPR/Cas9 technology was then used to generate InDels in the EBE of the OsSWEET13 promoter in MS14K to creat a new germplasm with three edited OsSWEET EBEs and broad-spectrum resistance against all Xoo strains tested. Collectively, our findings illustrate how to disarm TALE-S co-evolved loci to generate broad-spectrum resistance through the loss of effector-triggered susceptibility in plants.

Xanthomonas oryzae pv。米曲霉（白叶枯病），水稻细菌性枯萎病的致病剂，采用转录激活状效应（故事）以诱导表达OsSWEET家族的推定的糖转运蛋白基因，其功能在赋予疾病易感性（小号在水稻植物） 。为了工程化广谱细菌白叶枯病的抗性，我们使用了CRISPR / Cas9介导的基因编辑来破坏水稻cv中两个S基因OsSWEET11和OsSWEET14的TALE结合元件（EBE）。Kitaake，其中包含Xa25 / OsSWEET13的隐性抗性等位基因。工程水稻品系MS14K对大多数Xoo菌株表现出广谱抗性，只有少数例外，这表明兼容的菌株可能含有新的TALE。我们在兼容的Xoo菌株LN18和PXO61中确定了两个PthXo2样TALE，分别为Tal5 _LN18和Tal7 _PXO61，作为主要毒力因子，发现Xoo编码至少五种类型的PthXo2样效应子。考虑到PthXo2 / PthXo2.1靶向OsSWEET13进行转录激活，分析了3000个水稻品种的基因组中OsSWEET13启动子中的EBE变异，并鉴定了10个Xa25样单倍型。我们发现Tal5_LN18和Tal7 _PXO61结合OsSWEET13启动子中略有不同的EBE序列以激活其表达。然后，使用CRISPR / Cas9技术在MS14K中OsSWEET13启动子的EBE中生成InDels，以创建具有三个经过编辑的OsSWEET EBE和针对所有测试Xoo菌株的广谱抗性的新种质。总的来说，我们的发现说明了如何使TALE - S共同进化的基因座解除武装，从而通过失去效应子触发的植物敏感性而产生广谱抗性。