标题:Biallelic Editing of the LOB1 YPromoter via CRISPR/Cas9 Creates Canker-Resistant 'Duncan' Grapefruit

作者:Jia, Hongge, Omar, Ahmad A., Orbovic, Vladimir and Wang, Nian

摘要: Citrus canker caused by Xanthomonas citri subsp. citri is one of the most devastating citrus diseases worldwide. Generating disease-resistant citrus varieties is considered one of the most efficient and environmentally friendly measures for controlling canker. X. citri subsp. citri causes canker symptoms by inducing the expression of canker susceptibility gene LOB1 via PthA4, a transcription activator-like (TAL) effector, by binding to the effector binding element (EBE) in the promoter region. In previous studies, canker-resistant plants were generated by mutating the coding region or the EBE of LOB1. However, homozygous or biallelic canker-resistant plants have not been generated for commercial citrus varieties, such as grapefruit (Citrus paradisi), which usually contain two alleles of LOB1 and thus, have two types of LOB1 promoter sequences: TI LOBP and TII LOBP. Two different sgRNAs were used to target both EBE types. Both 35S promoter and Yao promoter were used to drive the expression of SpCas9p to modify EBEPthA4-LOBP in grapefruit. Using 'Duncan' grapefruit epicotyls as explants, 19 genome-edited grape-fruit plants were generated with one biallelic mutant line (#Dun(Yao)7). X. citri subsp. citri caused canker symptoms on wild-type and nonbiallelic mutant plants but not on #Dun(Yao)7. XccPthA4 mutant containing the designer TAL effector dLOB1.5, which recognizes a conserved sequence in both wild-type and #Dun(Yao)7, caused canker symptoms on both wild-type and #Dun(Yao)7. No off-target mutations were detected in #Dun(Yao)7. This study represents the first time that CRISPR-mediated genome editing has been successfully used to generate disease-resistant plants for 'Duncan' grapefruit, paving the way for using disease-resistant varieties to control canker.

文献注录:Jia, Hongge, Omar, Ahmad A., Orbovic, Vladimir and Wang, Nian. Biallelic Editing of the LOB1 YPromoter via CRISPR/Cas9 Creates Canker-Resistant 'Duncan' Grapefruit [J]. . 2022.

报/刊名:》,发表于2022

文献类型: [J]

页码: 页 / 共

Copyright© 2012 中国农业科学院柑桔研究所 All rights reserved.   网站备案号:渝ICP备05000267号   栏目访问量:次.     联系站长