中国油料作物学报2025，Vol.47Issue(2)：319-327,9.DOI:10.19802/j.issn.1007-9084.2023311
利用CRISPR/Cas9技术敲除BnERF019降低甘蓝型油菜对黑胫病的抗性
Genome editing of BnERF019 gene by CRISPR/Cas9 for reduced blackleg disease resistance in Brassica napus L.
摘要
Abstract
To assist in disease-resistant breeding,function of transcription factor BnERF019 was investigated,which is up-regulated after induction by oilseed rape(Brassica napus)black leg pathogen(Leptosphaeria biglobosa)for 96 h.By utilizing Golden Gate vector system,modified gene editing vector pHS-BnERF019 was constructed for developing multi-copy CRISPR/Cas9 system.Agrobacteria-mediated genetic transformation was employed to infect Westar hypocotyls.Consequently,28 regenerated seedlings with positive gene editing were obtained(positive rate as 32.18%).Among them,15 exhibited sequential edits in target BnaC07g13470D and 19 displayed sequential edits in target BnaA07g10270D.Additionally,18 seedlings with edited sequences for target BnaA09g30360D and 23 seed-lings with edited sequences for target BnaC05g18050D were obtained.Furthermore,3 regenerated seedlings had single target sites gene editing while 11 had double target sites gene editing;6 had 3 target sites gene editing and 8 had 4 target sites gene editing.Results of blackleg disease resistance identification showed that the resistance of re-generated seedlings with 4-copy gene editing was significantly weaker than those of wild-type seedlings.关键词
CRISPR/Cas9/甘蓝型油菜/转录因子/BnERF019/遗传转化Key words
CRISPR/Cas9/Brassica napus L./transcription factor/BnERF019/genetic transformation分类
农业科学引用本文复制引用
赵丽丽,皇甫海燕,郭晨,牟春雨,李子钦,潘勇,燕孟娇,魏玉龙,史志丹,王胜国..利用CRISPR/Cas9技术敲除BnERF019降低甘蓝型油菜对黑胫病的抗性[J].中国油料作物学报,2025,47(2):319-327,9.基金项目
国家自然科学基金(32160676) （32160676）
内蒙古自然科学基金(2021SHZR2732) （2021SHZR2732）
内蒙古农牧业科技创新基金(2021CXJJN05) （2021CXJJN05）
