[1]吴致君,卢莉,黎星辉,等.茶树AP2/ERF-B3类转录因子基因的克隆与表达特性分析[J].南京农业大学学报,2014,37(4):67-75.[doi:10.7685/j.issn.1000-2030.2014.04.010]
 WU Zhijun,LU Li,LI Xinghui,et al.Isolation and expression profiles analysis of AP2/ERF-B3 group transcription factor from Camellia sinensis[J].Journal of Nanjing Agricultural University,2014,37(4):67-75.[doi:10.7685/j.issn.1000-2030.2014.04.010]
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茶树AP2/ERF-B3类转录因子基因的克隆与表达特性分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
37卷
期数:
2014年4期
页码:
67-75
栏目:
出版日期:
2014-06-25

文章信息/Info

Title:
Isolation and expression profiles analysis of AP2/ERF-B3 group transcription factor from Camellia sinensis
作者:
吴致君1 卢莉2 黎星辉1 房婉萍1 周琳1 谭国飞3 庄静1
1. 南京农业大学园艺学院/茶叶科学研究所, 江苏 南京 210095;
2. 武夷学院茶与食品学院, 福建 武夷山 354300;
3. 南京农业大学作物遗传与种质创新国家重点实验室, 江苏 南京 210095
Author(s):
WU Zhijun1 LU Li2 LI Xinghui1 FANG Wanping1 ZHOU Lin1 TAN Guofei3 ZHUANG Jing1
1. College of Horticulture/Tea Research Institute, Nanjing Agricultural University, Nanjing 210095, China;
2. School of Tea and Food Science, Wuyi University, Wuyishan 354300, China;
3. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
关键词:
茶树转录因子AP2/ERF进化分析三级结构表达分析
Keywords:
Camellia sinensistranscription factorAP2/ERFphylogenetic analysisthree-dimensional structureexpression analysis
分类号:
S571.1
DOI:
10.7685/j.issn.1000-2030.2014.04.010
摘要:
植物在受到高温、低温、干旱和盐害等逆境胁迫时,ERF作为信号转导因子被诱导并调控其他抗逆基因的表达。研究茶树AP2/ERF-B3类转录因子在不同茶树品种间的分子特性、组织表达和逆境响应情况,将有助于了解AP2/ERF转录因子在茶树逆境调控中的作用。以2个茶树品种‘安吉白茶’和‘迎霜’为试验材料,通过RT-PCR方法分别从2种茶树的cDNA中克隆得到CsERF-B3基因。利用实时定量PCR技术检测该基因在茶树根、茎、叶、花各组织和4种非生物逆境胁迫处理(4 ℃低温、38 ℃高温、200 g·L-1 PEG干旱处理、200 mmol·L-1 NaCl)中的表达情况。结果表明:2种茶树中CsERF-B3基因全长均为639 bp,编码212个氨基酸,含有保守的AP2结合域,是植物典型的AP2/ERF家族转录因子;该转录因子属于AP2/ERF转录因子家族中的ERF亚族B3组;该转录因子是亲水性蛋白,无序化特征明显,并与拟南芥AtERF1具有相似的三级结构;该基因在茶树根中表达量最高,并且均能快速响应高温(38 ℃)、低温(4 ℃)和高盐(200 mmol·L-1 NaCl)等非生物逆境胁迫。结论:环境中常见非生物胁迫可诱导茶树中CsERF-B3基因的表达,表明该AP2/ERF-B3类转录因子在茶树非生物胁迫中起着重要调节作用。
Abstract:
In higher plant,ERF subfamily transcription factor involved in growth and development,signal transduction,and regulated the expression of related genes in order to improve the resistance to the abiotic stresses,such as high temperature,low temperature,drought and salt damage.Here,the molecular characterization,tissue expression and abiotic stresses response of the AP2/ERF-B3 group transcription factor in different tea varieties were investigated.The results will contribute to understanding the roles of AP2/ERF-B3 transcription factor in tea plant(Camellia sinensis).The CsERF-B3 genes,which encode to the AP2/ERF-B3 transcription factor,were cloned by RT-PCR method using cDNA as template,from two tea plant cultivars‘Anjibaicha’and‘Yingshuang’,respectively.Then,cDNA and deduced amino acid sequence,phylogenetic tree,physical and chemical characterization,hydrophilicity/hydrophobicity,molecular modeling and disordered residues were analyzed by related software.The expression profiles of the CsERF-B3 gene in the tissues of tea plants(roots,stems,leaves and flowers)were detected.The leaves,which were respectively treated with 2 h under 4 kinds of abiotic stresses(4 ℃ low temperature,38 ℃ high temperature,200 g·L-1 PEG and 200 mmol·L-1 NaCl),were also detected by the quantitative real-time PCR analysis.Then,the multiple comparisons and plotting were finished by using Excel program.The results of sequence alignments and phylogenetic analysis showed that lengths of CsERF-B3 genes from the two tea plant cultivars were 639 bp,encoding 212 amino acids,respectively.The transcription factor of CsERF-B3 between 110-145 amino acid sites contained the AP2 DNA binding domain where both contained a conservative WLG and YRG element.The CsERF-B3 of tea plants was a typical AP2/ERF family transcription factors in plants.Homologous phylogenetic tree showed that this ERF transcription factor was the closest relatives to AtERF1(AT4G17500.1)and AtERF2(AT5G47220.1)in Arabidopsis,they all belong to group B3,one group of the ERF subfamily transcription factors.The CsERF-B3 was hydrophilic protein and chemical properties were similar among related species.The CsERF-B3 and AtERF1 had similar three-dimension structure and disordered residues feature.The homologies of ABERF and YSERF were 92.1% and 93.7% compared with the DNA binding domain of crystal structure PDB ID:1gcc,respectively.The highest expression levels of the CsERF-B3 genes were found in the root of tea plant.The CsERF-B3 gene could respond quickly to high temperature(38 ℃),low temperature(4 ℃)and high-salinity(200 mmol·L-1 NaCl)treatments,respectively.The expression of CsERF-B3 gene was induced by abiotic stresses in tea plant,indicating that the transcription factor of AP2/ERF-B3 plays an important role in the regulation of abiotic stress in tea plant.

参考文献/References:

[1] 童启庆.茶树栽培学[M].北京:中国农业出版社, 2007:4-33[Tong Q Q.Tea Tree Cultivation[M].Beijing:China Agriculture Press, 2007:4-33(in Chinese)]
[2] 宛晓春.茶叶生物化学[M].北京:中国农业出版社, 2003:319-358[Wan X C.Tea Biochemistry[M].Beijing:China Agriculture Press, 2003:319-358(in Chinese)]
[3] 胡雲飞, 张玥, 张彩丽, 等.茶树根系内生真菌与根际土壤真菌的季节多样性分析[J].南京农业大学学报, 2013, 36(3):41-46.doi:10.7685/j.issn.1000-2030.2013.03.007[Hu Y F, Zhang Y, Zhang C L, et al.Diversity analysis of the endophytic fungi in root and fungi in rhizosphere soil of tea plant in different seasons[J].Journal of Nanjing Agricultural University, 2013, 36(3):41-46(in Chinese with English abstract)]
[4] 房婉萍, 张玥, 阮光兴, 等.茶树CsH 1 基因结构及其内含子信息分析[J].南京农业大学学报, 2012, 35(4):37-40.doi:10.7685/j.issn.1000-2030.2012.04.007[Fang W P, Zhang Y, Ruan G X, et al.Analysis of genomic structure and introns of CsH 1 gene from tea plant[J].Journal of Nanjing Agricultural University, 2012, 35(4):37-40(in Chinese with English abstract)]
[5] Riechmann J L, Heard J, Martin G, et al.Arabidopsis transcription factors:genome-wide comparative analysis among eukaryotes[J].Science, 2000, 290(5499):2105-2110
[6] Sakuma Y, Liu Q, 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
[7] Zhuang J, Cai B, Peng R H, et al.Genome-wide analysis of the AP2/ERF gene family in Populus trichocarpa[J].Biochem Biophys Res Commun, 2008, 371(3):468-474
[8] 张晗, 信月芝, 郭惠明, 等.CBF转录因子及其在植物抗冷反应中的作用[J].核农学报, 2006, 20(5):406-409[Zhang H, Xin Y Z, Guo H M, et al.CBF transcriptional factors and its roles in plant freezing tolerance[J].Journal of Nuclear Agricultural Sciences, 2006, 20(5):406-409(in Chinese with English abstract)]
[9] 阳文龙, 刘敬梅, 刘强, 等.高羊茅DREB类转录因子基因的分离及鉴定分析[J].核农学报, 2006, 20(3):187-192[Yang W L, Liu J M, Liu Q, et al.Isolation and characterization of a DREB-LIKE transcription factor gene from tall fescue[J].Journal of Nuclear Agricultural Sciences, 2006, 20(3):187-192(in Chinese with English abstract)]
[10] Zhuang J, Peng R H, Cheng Z M, et al.Genome-wide analysis of the putative AP2/ERF family genes in Vitis vinifera[J].Scientia Horticulturae, 2009, 123(1):73-81
[11] 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
[12] Li M Y, Wang F, Jiang Q, et al.Genome-wide analysis of the distribution of AP2/ERF transcription factors reveals duplication and elucidates their potential function in Chinese cabbage(Brassica rapa ssp.pekinensis)[J].Plant Molecular Biology Reporter, 2013, 31(4):1002-1011
[13] Tamura K, Peterson D, Peterson N, et al.MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J].Molecular Biology and Evolution, 2011, 28(10):2731-2739
[14] Schwede T, Kopp J, Guex N, et al.SWISS-MODEL:an automated protein homology-modeling server[J].Nucleic Acids Research, 2003, 31(13):3381-3385
[15] Prilusky J, Felder C E, Zeev-Ben-Mordehai T, et al.FoldIndex?:a simple tool to predict whether a given protein sequence is intrinsically unfolded[J].Bioinformatics, 2005, 21(16):3435-3438
[16] Wilkins M R, Gasteiger E, Bairoch A, et al.Protein identification and analysis tools in the ExPASy server[J].Methods Mol Biol, 1999, 112:531-552
[17] Pfaffl M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Research, 2001, 29(9):2002-2007
[18] Allen M D, Yamasaki K, Ohme-Takagi M, et al.A novel mode of DNA recognition by a β-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA[J].EMBO J, 1998, 17(18):5484-5496
[19] Fink A L.Natively unfolded proteins[J].Curr Opin Struct Biol, 2005, 15(1):35-41
[20] Fujimoto S Y, Ohta M, Usui A, et al.Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression[J].Plant Cell, 2000, 12(3):393-404
[21] Qin Q L, Liu J G, Zhang Z, et al.Isolation, optimization, and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza sativa L.[J].Molecular Breeding, 2007, 19(4):329-340
[22] 韦善君, 孙振元, 巨关升, 等.冷诱导基因转录因子CBF1的组成型表达对植物的抗寒性及生长发育的影响[J].核农学报, 2005, 19(6):465-468[Wei S J, Sun Z Y, Ju G S, et al.Effect of constituted expression of CBF1 transcriptional factor on plant cold tolerance, growth and development[J]. Journal of Nuclear Agricultural Sciences, 2005, 19(6):465-468(in Chinese with English abstract)]
[23] Gao M J, Allard G, Byass L, et al.Regulation and characterization of four CBF transcription factors from Brassica napus[J].Plant Molecular Biology, 2002, 49(5):459-471
[24] Berrocal-Lobo M, Molina A, Solano R.Constitutive expression of ethylene-response-factor 1 in Arabidopsis confers resistance to several necrotrophic fungi[J].Plant Journal, 2002, 29(1):23-32
[25] Lorenzo O, Piqueras R, Sanchez-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
[26] Berrocal-Lobo M, Molina A.Ethylene response factor 1 mediates Arabidopsis resistance to the soilborne fungus Fusarium oxysporum[J].Molecular Plant-Microbe Interactions, 2004, 17(7):763-770
[27] 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
[28] Solano R, Stepanova A, Chao Q M, et al.Nuclear events in ethylene signaling:a transcriptional cascade mediated by ethylene-insensitive 3 and ethylene-response-factor 1[J].Genes and Development, 1998, 12(23):3703-3714
[29] Zhu Z Q, An F Y, Feng Y, et al.Derepression of ethylene-stabilized transcription factors(EIN3/EIL1)mediates jasmonate and ethylene signaling synergy in Arabidopsis[J].Proc Natl Acad Sci USA, 2011, 108(30):12539-12544
[30] 庄静, 周熙荣, 孙超才, 等.油菜沪油15中AP2/ERF-B3亚族转录因子的克隆和生物信息学分析[J].分子细胞生物学报, 2008, 41(3):192-205[Zhuang J, Zhou X R, Sun C C, et al.Cloning and bioinformatic analyzing of transcription factor AP2/ERF-B3 subfamily genes from Brassica napus l.HUYOU 15[J].Journal of Molecular Cell Biology, 2008, 41(3):192-205(in Chinese with English abstract)]
[31] Zhang W, Zou A, Miao J, et al.LeERF-1 , a novel AP2/ERF family gene within the B3 subcluster, is down-regulated by light signals in Lithospermum erythrorhizon[J].Plant Biology, 2011, 13(2):343-348

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备注/Memo

备注/Memo:
收稿日期:2013-8-13。
基金项目:国家自然科学基金项目(31200520);江苏省自然科学基金项目(BK2012774);教育部高等学校博士点基金项目(20120097120031);国家博士后科学基金项目(2013M541686)
作者简介:吴致君,硕士研究生。
通讯作者:庄静,教授,博导,研究方向为茶树分子生物学,E-mail:zhuangjing@njau.edu.cn。
更新日期/Last Update: 1900-01-01