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.

参考文献

[1] Luo JL, Zhao N, Lu CM. Plant Trihelix transcription factors family. Hereditas (Beijing), 2012, 34(12): 1551–1560. 罗军玲, 赵娜, 卢长明. 植物Trihelix转录因子家族研究进展. 遗传, 2012, 34(12): 1551–1560.
[2] Kay SA, Keith B, Shinozaki K, Chye ML, Chua NH. The rice phytochrome gene: structure, autoregulated expression, and binding of GT-1 to a conserved site in the 5' upstream region. Plant Cell, 1989, 1(3): 351–360.
[3] Park HC, Kim ML, Kang YH, Jeon JM, Yoo JH, Kim MC, Park CY, Jeong JC, Moon BC, Lee JH, Yoon HW, Lee SH, Chung WS, Lim CO, Lee SY, Hong JC, Cho MJ. Pathogen- and NaCl-induced expression of the SCaM-4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant Physiol, 2004, 135(4): 2150–2161.
[4] Fang YJ, Xie KB, Hou X, Hu HH, Xiong LZ. Systematic analysis of GT factor family of rice reveals a novel subfamily involved in stress responses. Mol Genet Genom, 2010, 283(2): 157–169.
[5] Wang R, Hong GF, Han B. Transcript abundance of rml 1, encoding a putative GT1-like factor in rice, is up-regul?ated by Magnaporthe grisea and down-regulated by light. Gene, 2004, 324: 105–115.
[6] Wang XH, Li QT, Chen HW, Zhang WK, Ma B, Chen SY, Zhang JS. Trihelix transcription factor GT-4 mediates salt tolerance via interaction with TEM2 in Arabidopsis. BMC Plant Biol, 2014, 14(1): 339.
[7] Xie ZM, Zou HF, Lei G, Wei W, Zhou QY, Niu CF, Liao Y, Tian AG, Ma B, Zhang WK, Zhang JS, Chen SY. Soybean Trihelix transcription factors GmGT-2A and GmGT-2B improve plant tolerance to abiotic stresses in transgenic Arabidopsis. PLoS One, 2009, 4(9): e6898.
[8] Xi J, Qiu YJ, Du LQ, Poovaiah BW. Plant-specific trihelix transcription factor AtGT2L interacts with calcium/cal?modulin and responds to cold and salt stresses. Plant Sci, 2012, 185–186: 274–280.
[9] Li B, Jiang S, Yu X, Cheng C, Chen SX, Cheng YB, Yuan JS, Jiang DH, He P, Shan LB. Phosphorylation of Trihelix transcriptional repressor ASR3 by MAP KINASE4 negatively regulates Arabidopsis immunity. Plant Cell, 2015, 27(3): 839–856.
[10] Giuntoli B, Lee SC, Licausi F, Kosmacz M, Oosumi T, van Dongen JT, Bailey-Serres J, Perata P. A Trihelix DNA binding protein counterbalances hypoxia-responsive transcriptional activation in Arabidopsis. PLoS Biol, 2014, 12(9): e1001950.
[11] Yoo CY, Pence HE, Jin JB, Miura K, Gosney MJ, Hasegawa PM, Mickelbart MV. The Arabidopsis GTL1 transcription factor regulates water use efficiency and drought tolerance by modulating stomatal density via transrepression of SDD1. Plant Cell, 2010, 22(12): 4128–4141.
[12] Weng H, Yoo CY, Gosney MJ, Hasegawa PM, Mickelbart MV. Poplar GTL1 is a Ca2+/calmodulin-binding transcription factor that functions in plant water use efficiency and drought tolerance. PLoS One, 2012, 7(3): e32925.
[13] Griffith ME, Concei??o ADS, Smyth DR. PETAL LOSS gene regulates initiation and orientation of second whorl organs in the Arabidopsis flower. Development, 1999, 126(24): 5635–5644.
[14] Brewer PB, Howles PA, Dorian K, Griffith ME, Ishida T, Kaplan-Levy RN, Kilinc A, Smyth DR. PETAL LOSS, a trihelix transcription factor gene, regulates perianth architecture in the Arabidopsis flower. Development, 2004, 131(16): 4035–4045.
[15] O’Brien M, Kaplan-Levy RN, Quon T, Sappl PG, Smyth DR. PETAL LOSS, a trihelix transcription factor that represses growth in Arabidopsis thaliana, binds the energy-sensing SnRK1 kinase AKIN10. J Exp Bot, 2015, 66(9): 2475–2485.
[16] Tzafrir I, Pena-Muralla R, Dickerman A, Berg M, Rogers R, Hutchens S, Sweeney TC, McElver J, Aux G, Patton D, Meinke D. Identification of genes required for embryo development in Arabidopsis. Plant Physiol, 2004, 135(3): 1206–1220.
[17] Gao MJ, Lydiate DJ, Li X, Lui H, Gjetvaj B, Hegedus DD, Rozwadowski K. Repression of seed maturation genes by a Trihelix transcriptional repressor in Arabidopsis seedlings. Plant Cell, 2009, 21(1): 54–71.
[18] Willmann MR, Mehalick AJ, Packer RL, Jenik PD. MicroRNAs regulate the timing of embryo maturation in Arabidopsis. Plant Physiol, 2011, 155(4): 1871–1884.
[19] Kaplan-Levy RN, Brewer PB, Quon T, Smyth DR. The trihelix family of transcription factors-light, stress and development. Trends Plant Sci, 2012, 17(3): 163–171.
[20] Yu CY, Cai XF, Ye ZB, Li HX. Genome-wide identification and expression profiling analysis of trihelix gene family in tomato. Biochem Biophys Res Commun, 2015, 468(4): 653–659.
[21] Scott A, Wyatt S, Tsou PL, Robertson D, Allen NS. Model system for plant cell biology: GFP imaging in living onion epidermal cells. Biotechniques, 1999, 26(6): 1125, 1128–1132.
[22] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2?ΔΔCT method. Methods, 2001, 25(4): 402–408.
[23] Zhou HB, Li SF, Deng ZY, Wang XP, Chen T, Zhang JS, Chen SY, Ling HQ, Zhang AM, Wang DW, Zhang XQ. Molecular analysis of three new receptor-like kinase genes from hexaploid wheat and evidence for their participation in the wheat hypersensitive response to stripe rust fungus infection. Plant J, 2007, 52(3): 420–434.
[24] Wang GF, Wei XN, Fan RC, Zhou HB, Wang XP, Yu CM, Dong LL, Dong ZY, Wang XJ, Kang ZS, Ling HQ, Shen QH, Wang DW, Zhang XQ. Molecular analysis of common wheat genes encoding three types of cytosolic heat shock protein 90 (Hsp90): functional involvement of cytosolic Hsp90s in the control of wheat seedling growth and disease resistance. New Phytol, 2011, 191(2): 418–431.
[25] Bruun-Rasmussen M, Madsen CT, Jessing S, Albrechtsen M. Stability of barley stripe mosaic virus-induced gene silencing in barley. Mol Plant-Microbe Interact, 2007, 20(11): 1323–1331.
[26] Kaplan-Levy RN, Quon T, O'Brien M, Sappl PG, Smyth DR. Functional domains of the PETAL LOSS protein, a trihelix transcription factor that represses regional growth in Arabidopsis thaliana. Plant J, 2014, 79(3): 477–491.
[27] Ayadi M, Delaporte V, Li YF, Zhou DX. Analysis of GT-3a identifies a distinct subgroup of trihelix DNA-binding transcription factors in Arabidopsis. FEBS Lett, 2004, 562(1–3): 147–154.
[28] García-Cano E, Magori S, Sun Q, Ding ZH, Lazarowitz SG, Citovsky V. Interaction of Arabidopsis trihelix-do?main transcription factors VFP3 and VFP5 with Agrobacterium virulence protein VirF. PLoS One, 2015, 10(11): e0142128. [29] Maréchal E, Hiratsuka K, Delgado J, Nairn A, Qin J, Chait BT, Chua NH. Modulation of GT-1 DNA-binding activity by calcium-dependent phosphorylation. Plant Mol Biol, 1999, 40(3): 373–386.
[30] Hiratsuka K, Wu XD, Fukuzawa H, Chua NH. Molecular dissection of GT-1 from Arabidopsis. Plant Cell, 1994, 6(12): 1805–1813.
[31] Nagata T, Niyada E, Fujimoto N, Nagasaki Y, Noto K, Miyanoiri Y, Murata J, Hiratsuka K, Katahira M. Solution structures of the trihelix DNA-binding domains of the wild-type and a phosphomimetic mutant of Arabidopsis GT-1: mechanism for an increase in DNA-binding affinity through phosphorylation. Proteins, 2010, 78(14): 3033– 3047.
[32] Gourrierec JL, Li YF, Zhou DX. Transcriptional activation by Arabidopsis GT-1 may be through interaction with TFIIA-TBP-TATA complex. Plant J, 1999, 18(6): 663–668.
[33] Breuer C, Kawamura A, Ichikawa T, Tominaga-Wada R, Wada T, Kondou Y, Muto S, Matsui M, Sugimoto K. The Trihelix transcription factor GTL1 regulates ploidy-dependent cell growth in the Arabidopsis trichome. Plant Cell, 2009, 21(8): 2307–2322.
[34] Ji JH, Zhou YJ, Wu HH, Yang LM. Genome-wide analysis and functional prediction of the Trihelix transcription factor family in rice. Hereditas (Beijing), 2015, 37(12): 1228–1241.
纪剑辉, 周颖君, 吴贺贺, 杨立明. 水稻Trihelix转录因子家族全基因组分析及功能预测. 遗传, 2015, 37(12): 1228–1241.
[35] Lin ZW, Griffith ME, Li XR, Zhu ZF, Tan LB, Fu YC, Zhang WX, Wang XK, Xie DX, Sun CQ. Origin of seed shattering in rice (Oryza sativa L.). Planta, 2007, 226(1): 11–20.
[36] Li CB, Zhou AL, Sang T. Rice domestication by reducing shattering. Science, 2006, 311(5769): 1936–1939. [37] Li Y, Liu XD, Dong YM, Xie ZM, Chen SY. Cloning and functional analysis of the cotton Trihelix transcription factor GhGT29. Hereditas (Beijing), 2015, 37(12): 1218– 1227.
李月, 刘晓东, 董永梅, 谢宗铭, 陈受宜. 棉花Trihelix转录因子GhGT29基因的克隆及功能分析. 遗传, 2015, 37(12): 1218–1227.
[38] Green PJ, Kay SA, Chua NH. Sequence-specific interactions of a pea nuclear factor with light-responsive elements upstream of the rbcS-3A gene. EMBO J, 1987, 6(9): 2543–2549.

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

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

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	史晓黎,何伊琳,凌宏清. 小麦A基因组测序与进化研究进展[J]. 遗传, 2019, 41(9): 836-844.
[2]	孙兆庆, 闫波. 转录因子GATA6在心血管疾病中的作用及其调控机制[J]. 遗传, 2019, 41(5): 375-383.
[3]	于好强,孙福艾,冯文奇,路风中,李晚忱,付凤玲. 转录因子BES1/BZR1调控植物生长发育及抗逆性[J]. 遗传, 2019, 41(3): 206-214.
[4]	鞠君毅,赵权. γ-珠蛋白基因表达调控机制与临床应用[J]. 遗传, 2018, 40(6): 429-444.
[5]	丁庆倩,王小婷,胡利琴,齐欣,葛林豪,徐伟亚,徐兆师,周永斌,贾冠清,刁现民,闵东红,马有志,陈明. 谷子MYB类转录因子SiMYB42提高转基因拟南芥低氮胁迫耐性[J]. 遗传, 2018, 40(4): 327-338.
[6]	任岚,肖茹丹,张倩,娄晓敏,张昭军,方向东. KLF1和KLF9对K562细胞红系分化的协同调控作用[J]. 遗传, 2018, 40(11): 998-1006.
[7]	张玲, 何建波. GATA6在肝脏发育中的作用及调控机制[J]. 遗传, 2018, 40(1): 22-32.
[8]	岳敏, 杨禹, 郭改丽, 秦曦明. 哺乳动物生物钟的遗传和表观遗传研究进展[J]. 遗传, 2017, 39(12): 1122-1137.
[9]	郭文雅,崔艳梅,王婷婷,喻德跃,黄方. 野生大豆花发育相关基因GsLFY的功能研究[J]. 遗传, 2017, 39(1): 56-65.
[10]	向小华, 吴新儒, 晁江涛, 杨明磊, 杨帆, 陈果, 刘贯山, 王元英. 普通烟草WRKY基因家族的鉴定及表达分析[J]. 遗传, 2016, 38(9): 840-856.
[11]	李晓旭, 刘成, 李伟, 张增林, 高晓明, 周慧, 郭永峰. 番茄WOX转录因子家族的鉴定及其进化、表达分析[J]. 遗传, 2016, 38(5): 444-460.
[12]	杨明磊, 晁江涛, 王大伟, 胡军华, 吴华, 龚达平, 刘贯山. 烟草C2H2锌指蛋白转录因子家族成员的鉴定与表达分析[J]. 遗传, 2016, 38(4): 337-349.
[13]	翟亚男, 许泉, 郭亚, 吴强. 原钙粘蛋白基因簇调控区域中成簇的CTCF结合位点分析[J]. 遗传, 2016, 38(4): 323-336.
[14]	谷彦冰, 冀志蕊, 迟福梅, 乔壮, 徐成楠, 张俊祥, 周宗山, 董庆龙. 桃WRKY基因家族全基因组鉴定和表达分析[J]. 遗传, 2016, 38(3): 254-270.
[15]	马建辉, 仝豆豆, 张文利, 张黛静, 邵云, 杨云, 姜丽娜. 乌拉尔图小麦NAC转录因子的筛选与分析[J]. 遗传, 2016, 38(3): 243-253.