-
AP2(APETALA2)/ERF(ethylene responsive factor,ERF)转录因子超家族是植物中重要的转录因子类型,其共同的结构特征是在DNA结合区包含1个或多个由约60~70个氨基酸组成的高度保守的AP2/ERF结构域。根据结构域数目及其所结合的顺式作用元件的差异,AP2/ERF转录因子超家族可分为:AP2,乙烯反应因子,干旱反应元件结合蛋白(dehydration responsive element binding protein,DREB)和RAV 4个转录因子家族。ERFs是植物中特有的转录因子家族,研究证实ERF转录因子参与调控包括植物生长、发育在内的多个生物学过程[1-3]。另外,通过AP2/ERF结构域和PR基因启动子顺式作用元件GCC-box(AGCCGCC)的结合,ERF基因参与植物对环境刺激,尤其是对病原菌胁迫的响应。为了发现AP2/ERF转录因子在林木抗非生物胁迫以及抗病性中的作用,近年来,一些研究者对多年生木本模式植物——杨树Populus spp.的ERF和DREB转录因子进行了深入研究。Nanjo等[4]在脱水、高盐、高温、冷冻、脱落酸以及过氧化氢处理后的意大利杨Populus nigra var. italica EST文库鉴定到13个ERF/AP2转录因子,其中一些仅在特定的环境胁迫下特异表达。Zhuang等[5]对杨树AP2/ERF转录因子超家族进行了全基因组分析,鉴别出了91个ERF转录因子以及77个DREB转录因子。Maki等[6]发现欧洲黑杨Populus nigra ERF基因——PncERF1基因在室温生长的杨树细胞内不表达,仅对低温处理后产生响应表达。Li等[7]报道转番茄Solanum lycopersicum ERF转录因子的杂交杨Populus alba × Populus berolinensis获得了对盐胁迫的抗性,并且在200.0 mmol·L-1氯化钠下,转基因杨的干物质总量、树高,叶片中脯氨酸含量、钠离子(Na+)含量均高于非转基因杨。最近,应用数字基因表达方法(digital gene expression,DGE),Chen等[8]发现了2个盐胁迫后下调表达的小黑杨Populus simonii × Populus nigra AP2/ERF转录因子。此外,研究发现ERF转录因子也在杨树顶芽的休眠诱导和维持过程中表达[9]。近来,作者在溃疡病菌胁迫的多种杨树杂交无性系中发现1个特定ERF转录因子基因(ERF-18基因)的上调表达,预期该基因可能与杨树的抗逆反应相关。为深入揭示该基因的功能,本研究对中国优良杨树乡土树种——山海关杨Populus deltoides ‘Shanghaiguan’ ERF-18基因(PdERF-18基因)进行生物信息学分析,实时定量PCR(real-time QPCR)技术检测其在不同环境胁迫下的表达模式,以期阐明该基因的功能和抗逆反应机制,并为杨树抗逆遗传育种提供实验基础。
Expression patterns for a PdERF-18 response to different stresses in Populus deltoides 'Shanhaiguan'
-
摘要: 对一个溃疡病菌Botryosphaeria dothidea胁迫相关的山海关杨Populus deltoides ‘Shanghaiguan’乙烯反应因子(ethylene responsive factors, ERFs)基因——PdERF-18基因的系统发育、编码产物结构以及多种胁迫下的表达模式进行了分析。结果表明:PdERF-18基因编码产物属于ERF转录因子家族B-6类群, 具有植物ERF转录因子的典型结构, 但它与转录调控功能密切相关的AP2/ERF结构域第19位氨基酸位点发生替换, 推测PdERF-18基因可能具有不同于其他典型ERF转录因子的功能; 实时定量分析表明:溃疡病菌、根癌农杆菌Agrobacterium tumefaciens(Rhizobium radiobacter), 脱水、机械胁迫、盐胁迫和高温处理、茉莉酸以及水杨酸处理均可诱导PdERF-18基因的上调表达。其中溃疡病菌与茉莉酸处理以及根癌农杆菌与水杨酸处理下, 杨树PdERF-18基因分别具有相似的表达特征, 显示溃疡病菌、根癌农杆菌侵染的信号转导可能分别与茉莉酸/乙烯途径、水杨酸信号传导途径有关。PdERF-18基因可以对多种非生物、生物胁迫产生响应, PdERF-18基因有可能作为一种新的遗传资源而应用于杨树抗性遗传改良。Abstract: Ethylene responsive factors (ERFs), one kind of APETALA2(AP2)/ERF transcriptional factor that responds to environmental stimuli, especially to pathogen attacks, play an important role in plant-pathogen interactions. Recently, we noticed that one ERF-18 gene involved in the pathogenesis processes of the poplar stem canker disease of some poplar hybrid clones. However, the expression patterns of this gene response to other environmental stimuli are unclear. In this study, the phylogenesis, structure, and expression patterns of one Populus deltoides ERF gene (PdERF-18) were reported. Treatments including salicylic acid and jasmonic acid (JA) induction, Botryosphaeria dothidea and Agrobacterium tumefaciens (Rhizobium radiobacter) inoculation, mechanical stress, high temperature, salt stress, and dehydration stress were tested with real-time reverse transcription polymerase chain reaction (RT-qPCR). Results revealed a PdERF-18 coding for a B-6 member in the ERF family B. The 14th alanine and 19th aspartate residues in the AP2/ERF domain, typically characteristic of plant ERFs, were altered with an aspartate site substituted by a glutamate (an amino acid residue with stronger electronegativity than aspartate) in the coding product for the PdERF-18 gene implying that PdERF-18 might have some additional functions besides its resistance to a pathogen. RT-qPCR data of the diverse stresses revealed up-regulated expressed patterns in P. deltoides. Also, similar expression pattern responses for both B. dothidea and JA as well as responses to A. tumefaciens and salicylic acid were found. Thus, it was inferred that the JA/ethylene (ET) transduction pathway regulated gene expression in poplar canker pathogen attacks, but the SA pathway was involved in the responsiveness to poplar crown gall disease; therefore, it was deduced that PdERF-18 was a multi-functional transcriptional factor involved in many biotic and abiotic stresses.
-
-
[1] FENG Jianxun, LIU Di, PAN Yi, et al. An annotation update via cDNA sequence analysis and comprehensive profiling of developmental, hormonal or environmental responsiveness of the Arabidopsis AP2/EREBP transcription factor gene family[J]. Plant Mol Biol, 2005, 59(6):853-868. [2] GUTTERSON N, REUBER T L. Regulation of disease resistance pathways by AP2/ERF transcription factors[J]. Curr Opin Plant Biol, 2004, 7(4):465-471. [3] NAKANO T, SUZUKI K, FUJIMURA T, et al. Genome-wide analysis of the ERF gene family in Arabidopsis and rice[J]. Plant Physiol, 2006, 140(2):411-432. [4] NANJO T, FUTAMURA N, NISHIGUCHI M, et al. Characterization of full-length enriched expressed sequence tags of stress-treated poplar leaves[J]. Plant Cell Physiol, 2004, 45(12):1738-1748. [5] ZHUNAG Jing, CAI Bin, PENG Rihe, et al. Genome-wide analysis of the AP2/ERF gene family in Populus trichocarpa[J]. Biochem Biophys Res Commun, 2008, 371(3):468-474. [6] MAKI H, SATO M, OGAWA K, et al. Cloning and expression profile of an ERF gene isolated from cold-stressed poplar cells (Populus nigra)[J]. Cytol Int J Cytol, 2011, 76(1):11-18. [7] LI Yiliang, SU Xiaohua, ZHANG Bingyu, et al. Expression of jasmonic ethylene responsive factor gene in transgenic poplar tree leads to increased salt tolerance[J]. Tree Physiol, 2009, 29(2):273-279. [8] CHEN Su, JIANG Jing, LI Huiyu, et al. The salt-responsive transcriptome of Populus simonii×Populus nigra via DGE[J]. Gene, 2012, 504(2):203-212. [9] ROHDE A, RUTTINK T, HOSTYN V, et al. Gene expression during the induction, maintenance, and release of dormancy in apical buds of poplar[J]. J Exper Bot, 2007, 58(15/16):4047-4060. [10] CHANG Shujun, PURYEAR J, CAIRNEY J. A simple and efficient method for isolating RNA from pine trees[J]. Plant Mol Biol Rep, 1993, 11(2):113-116. [11] TAMURA K, PETERSON D, PETERSON N, et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28(10):2731-2739. [12] SAKUMA Y, LIU Qiang, DUBOUZET J G, et al. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration and cold inducible gene expression[J]. Biochem Biophys Res Commun, 2002, 290(3):998-1009. [13] EDGAR R C. MUSCLE:multiple sequence alignment with high accuracy and high throughput[J]. Nucl Acids Res, 2004, 32(5):1792-1797. [14] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and and the 2-⊿⊿CT method[J]. Methods, 2001, 25(4):402-408. [15] AGARWAL M, HAO Yujin, KAPOOR A, et al. A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance[J]. J Biol Chem, 2006, 281(49):37636-37645. [16] McGRATH K C, DOMBRECHT B, MANNERS J M, et al. Repressor and activator type ethylene response factors functioning in jasmonate signaling and disease resistance identified via a genome-wide screen of Arabidopsis transcription factor gene expression[J]. Plant Physiol, 2005, 139(2):949-959. [17] 郭琦, 王保垒, 王博文, 等.毛白杨PtDREB2A基因的克隆、表达及单核苷酸多态性分析[J].林业科学, 2011, 47(4):49-56. GUO Qi, WANG Baolei, WANG Bowen, et al. Isolation, expression and single nucleotide polymorphisms analysis of PtDREB2A in Populus tomentosa[J]. Sci Silv Sin, 2011, 47(4):49-56. [18] 刘士旺, 吴学龙, 郭泽建.拟南芥的抗病信号传导途径[J].植物病理学报, 2003, 33(2):104-111. LIU Shiwang, WU Xuelong, GUO Zejian. The signaling pathways of disease resistance in Arabidopsis[J]. Acta Phytopathol Sin, 2003, 33(2):104-111. [19] LIU Sixin, ANDERSON J A. Marker assisted evaluation of Fusarium head blight resistant wheat germplasm[J]. Crop Sci, 2003, 43(3):760-766. [20] THOMMA B P H J, EGGERMONT K, BROEKAERT W F, et al. Disease development of several fungi on Arabidopsis can be reduced by treatment with methyl jasmonate[J]. Plant Physiol Biochem, 2000, 38(5):421-427. [21] BERROCAL-LOBO M, MOLINA A. Ethylene response factor 1 mediates Arabidopsis resistance to soilborne fungus Fusarium oxysporum[J]. Mol Plant-Micr Int, 2004, 17(7):763-770. [22] 高海波, 沈应柏.水杨酸甲酯、苯骈噻唑及茉莉酸甲酯对合作杨防御物质的影响[J].西北林学院学报, 2007, 22(6):32-35. GAO Haibo, SHEN Yingbai. Effects of BTH, MeSA and MeJA on defensive components of Populus simonii×P. pyramidalis[J]. J Northwest For Univ, 2007, 22(6):32-35. [23] 张可文, 安钰, 胡增辉, 等.脂氧合酶、脱落酸与茉莉酸在合作杨损伤信号传递中的相互关系[J].林业科学研究, 2005, 18(3):300-304. ZHANG Kewen, AN Yu, HU Zenghui, et al. Relationship between lipoxygenase and ABA and JA in wounded signal transduction of heathy populus seedlings[J]. For Res, 2005, 18(3):300-304. [24] CHENG Qiang, ZHANG Bo, ZHUGE Qiang, et al. Expression profiles of two novel lipoxygenase genes in Populus deltoides[J]. Plant Sci, 2006, 170(6):1027-1035. [25] HOOYKAAS P J J, BEIJERSBERGEN. A G M. The virulence system of Agrobacterium tumefaciens[J]. Annu Rev Phytopathol, 1994, 32(1):157-179. [26] PRADEL K S, ULLRICH C I, CRUZ S S, et al. Symplastic continuity in Agrobacterium tumefaciens-induced tumors[J]. J Exp Bot, 1999, 50(331):183-192. [27] DITT R F, NESTER E W, COMAI L. Plant gene expression response to Agrobacterium tumefaciens[J]. Proc Nat Acad Sci USA, 2001, 98(19):10954-10959. [28] VEENA, JIANG Hongmei, DOERGE R W, et al. Transfer of T-DNA and Vir proteins to plant cells by Agrobacterium tumefaciens induces expression of host genes involved in mediating transformation and suppresses host defense gene expression[J]. Plant J, 2003, 35(2):219-236. [29] LORENZO O, PIQUERAS R, SáNCHEZ-SERRANO J J, et al. Ethylene-response-factor 1 integrates signals from ethylene and jasmonate pathways in plant defense[J]. Plant Cell, 2003, 15(1):165-178. [30] GU Yongqiang, WILDERMUTH M C, CHAKRAVARTHY S, et al. Tomato transcription factors Pti4, Pti5, and Pti6 activate defense responses when expressed in Arabidopsis[J]. Plant Cell, 2002, 14(4):817-831. -
链接本文:
https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.2014.05.009