摘要
为了探明类甜蛋白(thaumatin-like proteins,TLPs)基因与小麦抗叶锈病防御反应的相关性,本研究采用RT-PCR和电子克隆技术从叶锈菌侵染的TcLr19小麦叶片中获得了一个类甜蛋白基因,命名为TaLr19TLP1(GeneBank登录号:KJ764822)。生物信息学分析表明该基因包含一个长516 bp的开放阅读框(open reading frame,ORF),不含内含子,编码171个氨基酸残基,具有一个甜蛋白保守结构域(thaumatin,THN),氨基酸序列与其他物种类甜蛋白序列具有较高同源性。实时荧光定量PCR分析明确该基因表达受叶锈菌、脱落酸和水杨酸的诱导。接种叶锈菌后不同时间点,TaLr19TLP1基因在非亲和组合中表达早于亲和组合,且表达量明显高于亲和组合。另外,该基因表达具有组织特异性,在茎部表达量高于叶部和根部。Western杂交分析表明,TaLr19TLP1蛋白与小麦叶锈菌诱导具有相关性,与基因水平表达模式相一致。以上研究结果说明TaLr19TLP1基因参与小麦TcLr19小麦抗叶锈防御反应。
To understand the roles of thaumatin-like proteins （TLPs） gene in the response of wheat to leaf rust （Puccinia triticina） infection, one thaumatin-like proteins gene designated TaLr19TLP1 （GeneBank accession number： KJ764822） was obtained from TcLrl9 wheat leaves infected by leaf rust pathogen using RT-PCR and silico cloning methods. Bioinformatics analysis showed that TaLr19TLP1 contained a single opening reading frame of 516 bp encoding 171 amino acid-peptides without intron, and the deduced amino acids sequence of TaLr19TLP1 has high homology with pathogenesis-related protein 5 obtained from other species with a thaumatin THN domain. The expression profiles of the TaLr19TLP1 gene detected by real-time quantitative PCR showed that TaLr19TLP1 gene could be induced by Puccinia triticina, abscisic acid （ABA） and salicylic acid （SA）. TaLr19TLP1 was induced earlier and its expression level was higher in the incompatible interaction between wheat and Puccinia triticina than in compatible interactions. The tissue organ-specific expression pattern of the TaLr19TLP1 showed that its transcripts were detected abundantly in stems, and weakly in leaves and roots. Western blotting was conducted to confirm that TaLrl9TLP1 protein was induced by Puccinia triticina infection which is similar to the expression profiles at the nucleotide level. All these results indicated that TaLr19TLP1 may play an important role in wheat resistance to leaf rust pathogen attack.
出处
《分子植物育种》
CAS
CSCD
北大核心
2014年第5期867-874,共8页
Molecular Plant Breeding
基金
河北省自然科学基金(C2012204005)
国家重点基础研究发展计划资助基金(2013CB127700)共同资助
关键词
小麦
叶锈菌
类甜蛋白
表达分析
Wheat
Puccinia triticina
Thaumatin-like proteins
Expression analysis
