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为了研究普通小麦(TriticumaestivumL.)中是否有EDR1途径存在,根据拟南芥EDR1基因及其同源物设计了一对兼并性引物,用来分离小麦的EDR1同源物。以用小麦叶片RNA合成的cDNA为模板进行RT-PCR扩增,获得了代表小麦EDR1基因(命名为TaEDR1)的627bp长的cDNA片段(GenBank登录号:AY743662)。此后,通过RACE技术成功地获得了编码959个氨基酸的全长TaEDR1基因的cDNA序列。TaEDR1的氨基酸序列与大麦EDR1(标记为HvEDR1)有92%的相同。在TaEDR1的羧基末端有一个高度保守的丝氨酸/苏氨酸激酶催化功能域。因为存在一个推测的核定位基序,这个蛋白可能在细胞核中起作用。首次提供了证明普通小麦中存在EDR1同源物的分子生物学证据。用半定量RT-PCR方法研究了接种小麦白粉病菌[Blumeriagraminis(DC.)E.O.Speerf.sp.triticiEm.Marchal,Bgt]后叶片中TaEDR1基因的转录谱。结果表明,在接种白粉病菌后TaEDR1基因在叶片中的转录水平提高。组织特异性表达谱分析证明,小麦TaEDR1基因在叶片、茎、穗、根中均有表达。研究提示TaEDR1可能在小麦防卫应答反应中起作用。
In order to study whether there is EDR1 pathway in common wheat (Triticum aestivum L.), a pair of degenerate primers was designed based on the Arabidopsis EDR1 gene and its homologues to isolate wheat EDR1 homologues. A 627-bp cDNA fragment (GenBank accession number: AY743662) representing the wheat EDR1 gene (named TaEDR1) was obtained by RT-PCR amplification using cDNA synthesized from wheat leaf RNA as a template. Since then, the cDNA sequence encoding the full-length TaEDR1 gene of 959 amino acids has been successfully obtained by RACE technology. The amino acid sequence of TaEDR1 is 92% identical to that of barley EDR1 (labeled as HvEDR1). There is a highly conserved serine / threonine kinase catalytic domain at the carboxy terminus of TaEDR1. Because of the existence of a putative nuclear localization motif, this protein may play a role in the nucleus. For the first time, molecular biological evidence to demonstrate the presence of EDR1 homologs in common wheat is provided. The TaEDR1 gene transcriptional profile in leaf after inoculation of Blumeria graminis (DC.) E.O.peerf.sp. triticiEm. Marchal, Bgt was studied by semi-quantitative RT-PCR. The results showed that the transcription level of TaEDR1 gene in leaves increased after inoculation with powdery mildew. Tissue-specific expression profiling demonstrated that the wheat TaEDR1 gene was expressed in leaves, stems, spikes and roots. Research suggests that TaEDR1 may play a role in the defense response of wheat.