转录因子基因TuGTγ-3参与乌拉尔图小麦对条锈病的抗性

doi:10.16288/j.yczz.16-133

遗传 ›› 2016, Vol. 38 ›› Issue (12): 1090-1101.doi: 10.16288/j.yczz.16-133

转录因子基因TuGTγ-3参与乌拉尔图小麦对条锈病的抗性

丁刘军^{1, 2}，普明宇^{1, 2}，卫波¹，王献平¹，范仁春¹，张相岐¹

1. 中国科学院遗传与发育生物学研究所，植物细胞与染色体国家重点实验室，北京 100101；
2. 中国科学院大学，北京 100049

收稿日期:2016-04-18 修回日期:2016-07-10 出版日期:2016-12-20 发布日期:2016-09-14
通讯作者: 范仁春，助理研究员，研究方向：小麦抗病基因的分离与功能研究。E-mail: rcfan@genetics.ac.cn张相岐，研究员，研究方向：小麦重要基因克隆与分子育种。E-mail: xqzhang@genetics.ac.cn
作者简介:丁刘军，硕士研究生，专业方向：作物遗传育种学。E-mail: ljding@genetics.ac.cn
基金资助:
国家自然科学基金面上项目(编号：31471484)和国家重点基础研究发展计划(973计划)项目(编号：2013CB127703)资助

Transcription factor gene TuGTγ-3 is involved in the stripe rust resistance in Triticum urartu

Liujun Ding^{1, 2}, Mingyu Pu^{1, 2}, Bo Wei¹, Xianping Wang¹, Renchun Fan¹, Xiangqi Zhang¹

1. State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2016-04-18 Revised:2016-07-10 Online:2016-12-20 Published:2016-09-14
Supported by:
Supported by the National Natural Science Foundation of China (No. 31471484) and National Basic Research Program of China (973 Plan) ( No. 2013CB127703)

摘要/Abstract

摘要： 小麦条锈病是由条形柄锈菌小麦专化型(Puccinia striiformis West. f. sp. tritici Eriks.＆Henn., Pst)引起的一种严重的真菌病害，发掘新的抗条锈病相关基因对于小麦抗病育种和抗病机理研究都具有重要意义。Trihelix是植物特有的转录因子家族，参与调控生长发育、形态建成、胁迫应答等过程。迄今，小麦属Trihelix家族与抗条锈病相关的研究尚未见报道。本研究从乌拉尔图小麦(Triticum urartu Tum., 2n=2x=14, AA)中克隆了Trihelix家族GTγ亚家族中的一个基因，命名为TuGTγ-3。序列分析表明，TuGTγ-3基因具有完整的开放阅读框(ORF)，编码序列(CDS)全长1329 bp，编码442个氨基酸，推测其编码蛋白的分子量为50.31 kDa，理论等电点为6.12。生物信息学预测TuGTγ-3蛋白有一个单分型核定位信号(GLPMQKKMRYT)，没有信号肽和跨膜结构域。TuGTγ-3蛋白的保守trihelix结构域的氨基酸序列位置为Q115~R187，第四α-螺旋位置为F234~Y241，CC结构域的位置为K362~K436。二级结构分析显示，TuGTγ-3蛋白由43.89%的α-螺旋、9.51%的伸展链、9.95%的β-转角和36.65%的不规则卷曲构成。利用普通小麦的基因组数据库BLAST分析表明，TuGTγ-3被定位于5A染色体长臂上。瞬时表达实验显示，TuGTγ-3蛋白主要定位在细胞核中，但细胞质中也有少量分布。表达谱分析表明，TuGTγ-3基因在叶片中的表达量显著高于根和叶鞘，且受小麦条锈菌小种CYR32的诱导而强烈上调表达。进一步通过大麦条纹花叶病毒诱导的基因沉默(BSMV-VIGS)实验证明，转录因子TuGTγ-3正向调控了乌拉尔图小麦对条锈病的抗性。

关键词: 乌拉尔图小麦, 转录因子, TuGTγ-3基因, 条锈病抗性, BSMV-VIGS

Abstract: Wheat stripe rust, caused by Puccinia striiformis West. f. sp. tritici Eriks. ＆Henn. (Pst), is a serious fungal disease. Identification of new genes associate with stripe rust resistance is important for developing disease resistant wheat cultivars and studying the mechanism of disease resistance. Trihelix is a plant specific transcription factor family, which is involved in regulation of growth and development, morphogenesis, and response to stresses. So far, no study reports on the relationship between the Trihelix family and wheat stripe rust. In this study, a gene in the GTγ subfamily of Trihelix family, designated TuGTγ-3, was cloned from Triticum urartu Tum. (2n=2x=14, AA). The results of sequencing demonstrated that TuGTγ-3 gene consisted of a complete open reading frame (ORF), and its coding sequence was 1329 bp in length, which encoded a protein with 442 amino acids. The predicted molecular weight of this protein was 50.31 kDa and the theoretical isoelectric point was 6.12. Bioinformatic analysis revealed that TuGTγ-3 protein had a monopartite nuclear localization signal (GLPMQKKMRYT), and had neither transmembrane domain nor signal peptide. The conserved trihelix domain, the fourth α-helix and the CC domain were located in the regions of Q115?R187, F234?Y241 and K362?K436, respectively. Dissection of secondary structure showed that TuGTγ-3 protein comprised of 43.89% α-helix, 9.51% extended strand, 9.95% β-turn and 36.65% random coil structures. Based on the BLAST search against the genome database of common wheat from IWGSC, TuGTγ-3 was located on the long arm of chromosome 5A. Transient expression experiment using onion inner epidermal cell showed that the fusion protein TuGTγ-3-GFP distributed mainly in nuclear and slightly in cytoplasm. Expression profiles in different organs indicated that expression level of TuGTγ-3 was much higher in leaves than that in roots or leaf sheaths, and the expression in leaves was extremely up-regulated by infection of the Pst race CYR32. Furthermore, the BSMV-VIGS experiment demonstrated that the transcription factor TuGTγ-3 positively regulated resistance to stripe rust in T. urartu.

Key words: Triticum urartu, transcription factor, TuGTγ-3, stripe rust resistance, BSMV-VIGS

丁刘军，普明宇，卫波，王献平，范仁春，张相岐. 转录因子基因TuGTγ-3参与乌拉尔图小麦对条锈病的抗性[J]. 遗传, 2016, 38(12): 1090-1101.

Liujun Ding, Mingyu Pu, Bo Wei, Xianping Wang, Renchun Fan, Xiangqi Zhang. Transcription factor gene TuGTγ-3 is involved in the stripe rust resistance in Triticum urartu[J]. Hereditas(Beijing), 2016, 38(12): 1090-1101.

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转录因子基因TuGTγ-3参与乌拉尔图小麦对条锈病的抗性

Transcription factor gene TuGTγ-3 is involved in the stripe rust resistance in Triticum urartu

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