Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB1 promoter in citrus

文献类型： 外文期刊

第一作者： Peng, Aihong

作者： Peng, Aihong;Chen, Shanchun;Lei, Tiangang;Xu, Lanzhen;He, Yongrui;Yao, Lixiao;Zou, Xiuping;Peng, Aihong;Chen, Shanchun;Lei, Tiangang;Xu, Lanzhen;He, Yongrui;Yao, Lixiao;Zou, Xiuping;Peng, Aihong;Chen, Shanchun;Lei, Tiangang;Xu, Lanzhen;He, Yongrui;Wu, Liu;Yao, Lixiao;Zou, Xiuping

作者机构：

关键词： citrus canker;CsLOB1;genome editing;CRISPR;Cas9;resistance

期刊名称：PLANT BIOTECHNOLOGY JOURNAL （ 影响因子：9.803； 五年影响因子：9.555 ）

ISSN： 1467-7644

年卷期： 2017 年 15 卷 12 期

页码：

收录情况： SCI

摘要： Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is severely damaging to the global citrus industry. Targeted editing of host disease-susceptibility genes represents an interesting and potentially durable alternative in plant breeding for resistance. Here, we report improvement of citrus canker resistance through CRISPR/Cas9-targeted modification of the susceptibility gene CsLOB1 promoter in citrus. Wanjincheng orange (Citrus sinensis Osbeck) harbours at least three copies of the CsLOB1(G) allele and one copy of the CsLOB1(-) allele. The promoter of both alleles contains the effector binding element (EBEPthA4), which is recognized by the main effector PthA4 of Xcc to activate CsLOB1 expression to promote citrus canker development. Five pCas9/CsLOB1sgRNA constructs were designed to modify the EBEPthA4 of the CsLOB1 promoter in Wanjincheng orange. Among these constructs, mutation rates were 11.5%-64.7%. Homozygous mutants were generated directly from citrus explants. Sixteen lines that harboured EBEPthA4 modifications were identified from 38 mutant plants. Four mutation lines (S2-5, S2-6, S2-12 and S5-13), in which promoter editing disrupted CsLOB1 induction in response to Xcc infection, showed enhanced resistance to citrus canker compared with the wild type. No canker symptoms were observed in the S2-6 and S5-13 lines. Promoter editing of CsLOB1(G) alone was sufficient to enhance citrus canker resistance in Wanjincheng orange. Deletion of the entire EBEPthA4 sequence from both CsLOB1 alleles conferred a high degree of resistance to citrus canker. The results demonstrate that CRISPR/Cas9-mediated promoter editing of CsLOB1 is an efficient strategy for generation of canker-resistant citrus cultivars.

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