水稻泛素结合酶基因家族的生物信息学与表达分析

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  • 福建农林大学 功能基因组学研究中心, 福州 350002;
*通讯联系人, E-mail: gdlufafu@163.com

收稿日期: 2015-11-30

  修回日期: 2016-03-16

  网络出版日期: 2016-05-10

基金资助

国家科技支撑计划资助项目(2013BAD19B03)

Bioinformatic and Expression Analysis of Rice Ubiquitin-conjugating Enzyme Gene Family

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  • The Functional Genomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
*Corresponding author, E-mail:gdlufafu@163.com

Received date: 2015-11-30

  Revised date: 2016-03-16

  Online published: 2016-05-10

摘要

泛素/蛋白酶体系统在植物的生长发育、形态建成和抗病反应等过程中起着重要的作用。近年的研究表明,某些病原菌能够模拟寄主植物泛素/蛋白酶体系统组分,从而达到利用该系统为病原菌服务的目的。泛素结合酶是泛素化反应过程中的第二个酶,对植物泛素/蛋白酶体系统的正常运行不可或缺。已有的研究表明,水稻基因组数据库中存在48个预测的泛素结合酶基因。为了初步揭示这些泛素结合酶基因在植物抗病防御反应中的功能,研究其与植物抗病性的关系。本研究通过生物信息学、RNA-seq和qRT-PCR的方法,分析了水稻泛素结合酶基因家族的特征及其表达模式。系统进化树分析表明,48个水稻泛素结合酶基因可分为3个大组,总共7个亚组。蛋白结构域分析表明,水稻泛素结合酶基因主要由一个泛素结合酶催化结构域组成。电子表达谱分析表明,大多数水稻泛素结合酶基因能被稻瘟病菌诱导表达。启动子区顺式作用元件分析表明,4个抗病相关顺式作用元件和1个过敏性反应相关顺式作用元件在48个水稻泛素结合酶基因的启动子区有很高的分布。稻瘟病菌接种亲和性和非亲和性水稻单基因系的RNA-seq结果表明,处理36 h读取到的水稻泛素结合酶基因为44个,其中高表达基因数量超过读取到的泛素结合酶基因总数的50%。qRT-PCR分析结果表明稻瘟病菌的侵染在亲和和非亲和组合中都能诱导部分水稻泛素结合酶基因的表达。在非亲和组合中水稻泛素结合酶基因的表达倾向于受到抑制。

本文引用格式

刘鑫, 张恒, 阚虎飞, 周立帅, 黄昊, 宋林林, 翟焕趁, 张君, 鲁国东 . 水稻泛素结合酶基因家族的生物信息学与表达分析[J]. 中国水稻科学, 2016 , 30(3) : 223 -231 . DOI: 10.16819/j.1001-7216.2016.5177

Abstract

The ubiquitin / proteasome system plays an important role in plant growth and development, morphogenesis and disease resistance. Recent studies have shown that some pathogens can mimic the host plant ubiquitin / proteasome system components to achieve their own purposes. Ubiquitin-conjugating enzyme is the second enzyme in the ubiquitination process and is indispensable for the plant ubiquitin/proteasome system. Previous studies showed that there are 48 predicted ubiquitin-conjugating enzyme genes in rice genome. In order to preliminarily elucidate the functions of rice ubiquitin-conjugating enzyme genes in plant disease resistance, bioinformatic, RNA-seq and qRT-PCR methods were used to analyze characteristics and expression patterns of rice ubiquitin-conjugating enzyme gene family. Phylogenetic tree analyses indicate that the 48 rice ubiquitin-conjugating enzyme genes can be divided into 3 groups, 7 sub-groups in total. Protein domain analysis showed that ubiquitin-conjugating enzyme genes mainly consist of a big ubiquitin-conjugating enzyme catalytic domain. Expression analysis in silico suggested that most of the rice ubiquitin-conjugating enzymes can be induced by blast fungus infection. Plant cis-acting elements analysis indicated that four pathogen resistance cis-acting elements and one hypersensitivity reaction cis-acting element have high distribution in the promoter region of the 48 rice ubiquitin-conjugating enzyme genes. RNA-seq data from compatible and incompatible monogenic rice after rice blast fungus infection showed that 44 rice ubiquitin-conjugating enzyme genes were expressed at 36 hours after treatment, among which more than 50% were highly expressed genes. qRT-PCR analysis showed that expression of some ubiquitin-conjugating enzyme genes can be induced by the inoculation of rice blast fungus both in compatible and incompatible monogenic rice. However, in incompatible rice the expression of rice ubiquitin-conjugating enzyme genes tends to be inhibited after rice blast fungus inoculation.

参考文献

[1] Hershko A.The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle.Cell Death Differ, 2005, 12: 1191-1197.
[2] Dielen A, Badaoui S, Candresse T, et al.The ubiquitin/26S proteasome system in plant-pathogen interactions: A never-ending hide-and-seek game.Mol Plant Pathol, 2010, 11(2): 293-308.
[3] Zeng L R, Miguel E V, Zhu T, et al.Ubiquitination-mediated protein degradation and modification: An emerging theme in plant-microbe interactions.Cell Res, 2006, 16: 423-426.
[4] Stone S L.The role of ubiquitin and the 26S proteasome in plant abiotic stress signaling.Front Plant Sci, 2014, 5.
[5] Marino D, Peeters N, Rivas S.Ubiquitination during plant immune signaling.Plant Physiol, 2012, 160(1): 15-27.
[6] Smalle J, Vierstra R D.The ubiquitin 26S proteasome proteolytic pathway.Annu Rev Plant Biol, 2004, 55: 555-590.
[7] Madden L V, Wheelis M.The threat of plant pathogens as weapons against U.S. crops.Annu Rev Phytopathol, 2003, 41(4): 155-176.
[8] Zeng L R, Qu S H, Bordeos A, et al.Spotted leaf11, a negative regulator of plant cell death and defense, encodes a U-Box/Armadillo repeat protein endowed with E3 ubiquitin ligase activity.Plant Cell, 2004, 16: 2795-2808.
[9] Park C, Chen S B, Shirsekar G, et al.The Magnaporthe oryzae effector AvrPiz-t targets the RING E3 ubiquitin ligase APIP6 to suppress pathogen-associated molecular pattern-triggered immunity in rice.Plant Cell, 2012, 24: 4748-4762.
[10] 蒋春苗, 黄兴奇, 付坚, 等.疣粒野生稻泛素结合酶基因的全长cDNA序列克隆与分析. 作物学报,2012,38(5): 808-813.
[10] Jiang C M, Huang X Q, Fu J, et al.Cloning and analysis on full-Length cDNA sequence of ubiquitin-conjugating enzyme gene from Oryza meyeriana Baill.Acta Agron Sin, 2012, 38(5): 808-813. (in Chinese with English abstract)
[11] 胡婷丽, 李魏, 刘雄伦, 等. 泛素化在植物抗病中的作用. 微生物学通报, 2014, 41(6): 1175-1179.
[11] Hu T L, Li W, Liu X L, et al.The role of ubiquitination in plant disease resistance.Microbiol China, 2014, 41(6): 1175-1179. (in Chinese with English abstract)
[12] 杨玖霞, 张浩, 王志龙, 等. E3泛素连接酶调控植物抗病分子机理研究进展. 植物保护,2015,41(4): 1-8.
[12] Yang J X, Zhang H, Wang Z L, et al.Recent progresses in the regulation mechanism of E3 ligases in plant disease resistance.Plant Protect, 2015, 41(4): 1-8. (in Chinese with English abstract)
[13] 林艺娟. OsRBCS和OsUBC2在水稻抗病防御中的作用机制. 福州:福建农林大学,2014.
[13] Lin Y J.The mechanism of OsRBCs and OsUBC2 in rice disease resistance. Fuzhou: Fujian Agriculture and Forestry University, 2014.(in Chinese with English abstract)
[14] Hansol B, Woo T K.Classification and interaction modes of 40 rice E2 ubiquitin-conjugating enzymes with 17 rice ARM-U-box E3 ubiquitin ligases.Biochem Biophys Res Commun, 2014, 444: 575-580.
[15] E Z G, Zhang Y P, Li T T, et al. Characterization of the ubiquitin-conjugating enzyme gene family in rice and evaluation of expression profiles under abiotic stresses and hormone treatments.PLoS ONE, 2015, 10(4).
[16] Xu L, Ménard R, Berr A, et al.The E2 ubiquitin-conjugating enzymes, AtUBC1 and AtUBC2, play redundant roles and are involved in activation of FLC expression and repression of flowering in Arabidopsis thaliana.Plant J, 2009, 57(2): 279-288.
[17] Cui F, Liu L J, Zhao Q Z, et al.Arabidopsis ubiquitin conjugase UBC32 is an ERAD component that functions in brassinosteroid-mediated salt stress tolerance.Plant Cell, 2012, 24(1): 233-244.
[18] Chung E, Cho C W, So H A, et al.Overexpression of VrUBC1, a mung bean E2 ubiquitin-conjugating enzyme, enhances osmotic stress tolerance in Arabidopsis.PLoS ONE, 2013, 8(6).
[19] Wan X R, Mo A Q, Liu S, et al.Constitutive expression of a peanut ubiquitin-conjugating enzyme gene in Arabidopsis confers improved water-stress tolerance through regulation of stress-responsive gene expression.J Biosci Bioengin, 2011, 111(4): 478-484.
[20] Zhou G A, Chang R Z, Qiu L J.Overexpression of soybean ubiquitin-conjugating enzyme gene GmUBC2 confers enhanced drought and salt tolerance through modulating abiotic stress-responsive gene expression in Arabidopsis.Plant Mol Biol, 2010, 72(4-5): 357-367.
[21] Jeon E H, Pak J H, Kim M J, et al.Ectopic expression of ubiquitin-conjugating enzyme gene from wild rice, OgUBC1, confers resistance against UV-B radiation and Botrytis infection in Arabidopsis thaliana.Biochem Biophys Res Commun, 2012, 427(2): 309-314.
[22] Kawahara Y, Bastide M D L, Hamilton J P, et al. Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data.Rice, 2013, 6: 4.
[23] Finn R D, Bateman A, Clements J, et al. The Pfam protein families database. Nucl Acids Res, 2014, Database Issue 42: D222-D230.
[24] Higo K, Ugawa Y, Iwamoto M,et al.Plant cis-acting regulatory DNA elements (PLACE) database.Nucl Acids Res, 1999, 27: 297-300.
[25] Prestridge D S.SIGNAL SCAN: A computer program that scans DNA sequences for eukaryotic transcriptional elements.Computer Appl Biosci, 1991, 7: 203-206.
[26] de Hoon M J L, Imoto S, Nolan J, et al. Open source clustering software.Bioinformatics, 2004, 20(9): 1453-1454.
[27] Mortazavi A, Williams B A, Mccue, et al. Mapping and quantifying mammalian transcriptomes by RNA-Seq.Nat Methods, 2008, 5(7): 621-628.
[28] van Demark A P, Hofmann R M, Tsui C, et al. Molecular insights into polyubiquitin chain assembly: Crystal structure of the Mms2/Ubc13 heterodimer.Cell, 2001, 105(6): 711-720.
[29] Andersen P L, Zhou H, Pastushok L, et al.Distinct regulation of Ubc13 functions by the two ubiquitin-conjugating enzyme variants Mms2 and Uev1A.J Cell Biol, 2005, 170(5): 745-755.
[30] Ye Y, Rape M.Building ubiquitin chains: E2 enzymes at work.Nat Rev Mol Cell Biol, 2009, 10(11): 755-764.
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