白粉菌诱导的小麦品种Brock的差异表达基因解析  

Analysis on differentially expressed genes of wheat Brock inoculated by Blumeria gramini f. sp. tritici

作　　者：王艳红[1,2] 肖莹[1,2] 郑舒扬 刘晓颖[1,2] 王振英[1,2] 

机构地区：[1]天津师范大学生命科学学院,天津300387 [2]天津师范大学天津市动植物抗性重点实验室,天津300387

出　　处：《天津师范大学学报（自然科学版）》2015年第2期71-76,93,共7页Journal of Tianjin Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目(31071671);天津市应用基础与前沿技术研究计划青年基金资助项目(14JCQNJC14900)

摘　　要：为了解抗病小麦品种Brock对白粉菌的抗性机制,采用m RNA差异显示技术(DDRT-PCR)和反向Northern杂交技术分析小麦Brock在白粉菌诱导下的差异表达基因.对照组共获得5 072个片段,诱导组获得5 366个片段,反向Northern杂交筛选后,获得94个差异表达片段.基因测序和Blast比对后,共有71个片段得到功能注释,涉及抗病防御相关基因、能量代谢相关基因、转录因子、次级代谢相关基因以及信号转导基因等.选取与RPP13同源的片段7和与MYB44同源的片段10进行半定量表达模式分析,发现白粉菌诱导后这2个基因的表达明显上调,在染菌8 h后达到峰值,表明二者可能参与Brock白粉菌侵染的早期应答反应.To investigate the resistant mechanisms to Blumeria gramini f. sp. tritici （Bgt）in disease-resistant wheat Brock, differentially expressed genes of wheat Brock induced by Bgt were analyzed by mRNA differential display reverse transcrip- tion PCR （DDRT-PCR） and reverse Northern hybridization technique. A total of 5 366 highly reproducible fragments in the Bgt-inoculated Brock and 5 072 fragments in the control were obtained separately. 94 differential expression fragments （DEFs） were produced after reverse Northern Blot screening. After gene sequencing and comparison by the method of Basic Local Alignment Search Tool （BLAST）, the gene annotation of 71 fragments were conducted, which shared significant simi- larities with genes involved in disease defense, energy metabolism, transcription factor, signal transduction, and secondary metabolism, et al. DEF7 was 94% similar to disease resistance gene RPP13 and DEF10 was 98% similar to transcription fac- tor gene MYB44. These two DEFs were selected and their transcriptional levels were assayed by the method of semiquantita- tive RT-PCR in Brock. The result showed that the expression levels of DEF7 and DEF10 were up-regulated obviously, and reached the maximums after inoculation of 8 h. This suggested that DEF7 and DEF10 might be involved in the early responses in Brock resistance to Bgt.

关 键 词：禾布氏白粉菌 差异表达基因 MRNA差异显示技术 反向Northern杂交 抗病基因 

分 类 号：Q945.8[农业科学—植物病理学] S512[生物学—植物学]