黑胫病菌侵染过程中油菜响应基因的表达分析  

Gene Expression Analysis of Brassica napus in Response to Leptosphaeria biglobosa Infection Progress

作　　者：郝丽芬[1,2] 燕孟娇 房永雨 史志丹 李子钦 韩冰 HAO Lifen;YAN Mengjiao;FANG Yongyu;SHI Zhidan;LI Ziqin;HAN Bing(College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010011, China;Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China)

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010011 [2]内蒙古自治区农牧业科学院,呼和浩特010031

出　　处：《西北植物学报》2021年第2期197-211,共15页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家重点研发计划(2018YFD0200907);内蒙古科技计划(2019GG343);内蒙古科技成果转化专项(2019CG014)。

摘　　要：油菜黑胫病是造成油菜产量损失的病害之一,致病菌为Leptosphaeria biglobosa。该研究采用形态学观察和转录组测序技术,分析油菜接种病原菌Leptosphaeria biglobosa 4、12、24、36、48和96 h后的表型及基因表达变化情况,以探讨响应死体营养型真菌L.biglobosa侵染时油菜的防御反应及抗病机理,为揭示油菜与L.biglobosa互作的分子机制提供理论依据,并为培育油菜抗病品种积累了基因资源信息。结果显示:(1)接种4~96 h,叶片病斑逐渐扩大,病原菌侵染48~96 h后形成菌丝网。(2)通过RNA-Seq测序,在L.biglobosa侵染油菜的不同时间点(4、12、24、36、48和96 h)分别得到3384、2270、3802、5811、6155和7153个差异表达基因。(3)15个油菜差异表达基因的qRT-PCR检测表达水平与转录组测序结果基本一致。(4)利用短时间序列聚类和KEGG富集分析差异表达基因,结果发现植物病原菌互作、蛋白激酶、茉莉酸/乙烯/水杨酸和芥子油苷合成途径中的基因被强烈诱导表达,而且基因表达呈动态变化趋势。Blackleg caused by Leptosphaeria biglobosa,is one of the damaging diseases of oilseed rape(Brassica napus).Based on histological observation and RNA-Seq,we analyzed the phenotypes and gene expression changes of oilseed rape after inoculation with L.biglobosa for 4,12,24,36,48 and 96 h to discuss the defense response and disease resistance mechanism of oilseed rape in response to the infection of L.biglobosa,provided a theoretical basis for comprehensively revealing the molecular mechanism of interaction between oilseed rape and L.biglobosa,and accumulated genetic information for breeding the disease resistance varieties of oilseed rape.The results showed that:(1)after inoculation for 4-96 h,the leaf spot gradually expanded,and the L.biglobosa formed mycelium within 48-96 h.(2)By RNA-Seq,a number of 3384,2270,3802,5811,6155,7153 differentially expressed genes(DEGs)were obtained at 4,12,24,36,48 and 96 h of L.biglobosa infection on oilseed rape.(3)qRT-PCR results showed that the levels of the 15 differentially expressed genes in oilseed rape were basically consistent with the results of RNA-Seq.(4)The differentially expressed genes were analyzed by short time series clustering and KEGG enrichment analysis.The results showed that the genes in the pathways of plant and L.biglobosa interaction,protein kinase,jasmonate/ethylene/salicylic acid signaling,and glucosinolate biosynthesis were strongly activated,and their gene expression showed dynamic change trend.

关 键 词：差异表达基因(DEGs) Leptosphaeria biglobosa 植物-病原菌互作 MAPK 茉莉酸/乙烯/水杨酸 芥子油苷合成 

分 类 号：Q786[生物学—分子生物学] Q789