Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

ZHAO Tian-tian; LIANG Li-song; MA Qing-hua; WANG Gui-xi

Volume 29 Issue 2

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Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

1.
State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Silviculture of State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Received Date: 2014-12-03
Accepted Date: 2015-11-24

Abstract

[Objective]To analyze the sequence features and expression rules of WRKY gene from Corylus heterophylla Fisch. [Method]The gene was cloned by RACE-PCR. Quantitative real-time PCR was used in analyzing gene expression in various tissues and different abiotic stresses, including cold, high salinity and drought. [Result]The cDNA of WRKY is 1 342 bp in length, including a complete open reading frame (ORF) of 963 bp encoding a protein of 320 amino acids, designated as ChWRKY28. Phylogeny tree results showed ChWRKY28 was much closer to AtWRKY28 from Arabidopsis thaliana and PtrWRKY93 from Populus trichocarpa, generated 49% and 60% amino acids similarity. Spatial expression analyses demonstrated that the expression level of ChWRKY28 was higher in stem than that in male anthotaxy and floral buds which indicating tissue-specific expression. ChWRKY28 was clearly induced by cold, high salinity and drought, but that the expression tendency were evidently different of this gene. The subcellular localization analysis showd that ChWRKY28 protein was targeted to the nucleus. [Conclusion]This study indicated that ChWRKY28 gene may be involved in response to abiotic stress signal transduction pathway.
- Corylus heterophylla Fisch,
- transcription factors,
- clone,
- subcellular localization,
- expression

References

[1]	马庆华,王贵禧,梁维坚,等.我国榛属植物种质资源的研究、利用与创新[J]. 果树学报,2013,30(1):159-164.
[2]	Mehlenbacher S A. Hazelnuts. Genetic resources in temperate fruit and nut crops[J]. Acta Horticulturae,1991,290:791-838.
[3]	Liu J,Cheng Y,Liu C,et al. Temporal changes of disodium fluorescein transport in hazelnut during fruit development stage[J]. Sci Hortic Amsterdam,2013,150:348-353.
[4]	Coyne C J, Mehlenbacher S A, Smith D C. Sources of resistance to Eastern Filbert Blight in hazelnut [J]. Journal of the American Society for Horticultural Science,1998,123(2):253-257.
[5]	吕跃东,董凤祥,王贵禧,等. 平欧杂交榛抗寒性综合评价体系的建立与应用[J]. 林业科学,2008,44(9):31-35.
[6]	彭立新,王明启,梁维坚,等. 榛属(Corylus L.)植物抗寒性研究[J]. 吉林林学院学报,1994,10(3):166-170.
[7]	赵天田,陈新,刘庆忠,等.平榛NAC转录因子的分离及表达特性分析[J]. 植物遗传资源学报,2012,13(2):265-270.
[8]	王磊,高晓清,朱苓华,等.植物WRKY转录因子家族基因抗病相关功能的研究进展[J]. 植物遗传资源学报,2011,12(1):80-85.
[9]	Wang H H,Hao J J,Chen X J,et al. Overexpression of rice WRKY89 enhances ultraviolet B tolerance and disease resistance in rice plants[J]. Plant Molecular Biology,2007,65:799-815.
[10]	Ulker B,Somssich I E. WRKY transcription factors:from DNA binding towards biological function[J]. Current Opinion in Plant Biology,2004,7(5):491-498.
[11]	Dong J,Chen C,Chen Z. Expression profiles of the Arabidopsis WRKY gene superfamily during plant defense response [J]. Plant Molecular Biology,2003,51 (1):21-37.
[12]	Eulgem T,Somssich I E. Networks of WRKY transcription factors in defense signaling [J]. Current Opinion in Plant Biology,2007,10:366-371.
[13]	Ishiguro S,Nakamura K. Characterization of a cDNA encoding a novel DNA-binding protein,SPF1,that recognizes SP8 sequences in the 50 upstream regions of genes coding for sporamin and beta-amylase from sweet potato [J]. Mol Gen Genet,1994,244(6):563-571.
[14]	Seki M,Narusaka M,Ishida J,et al. Monitoring the expression profiles of 7000 Arabidopsis genes under drought,cold and high-salinity stresses using a full-length cDNA microarray[J]. The Plant Journal,2002,31 (3):279-292.
[15]	Jiang Y,Deyholos M. Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses [J]. Plant Molecular Biology,2009,69 (1-2):91-105.
[16]	Chen X,Zhang J,Liu Q Z,et al. Transcriptome Sequencing and Identification of Cold Tolerance Genes in Hardy Corylus Species (C. heterophylla Fisch) Floral Buds[J]. PLoS One,2013,9(9):e108604.
[17]	Wang L Q,Wang C,Wang D Y,et al. Molecular characterization and transcript profiling of NAC genes in response to abiotic stress in Tamarix hispida[J]. Tree Genetics & Genomes,2014,10(1):157-171.
[18]	Chang S,Puryear J,Cairney J. A simple and eficient method for isolating RNA from pine trees [J]. Plant Molecular Biol Reporter,1993,11(2):113-116.
[19]	Zhang Y,Feng J C. Identification and characterization of the grape WRKY family[J]. Biomed Research International,2014,787680.
[20]	Wang C,Deng P Y,Chen L L,et al. A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco[J]. PLoS One,2013,8(6):e65120.
[21]	Jiang Y Z,Duan Y J,Yin J,et al. Genome-wide identification and characterization of the Populus WRKY transcription factor family and analysis of their expression in response to biotic and abiotic stresses[J]. J Exp Bot,2014,65:6629-6644.
[22]	罗昌国,渠慎春,张计育,等.湖北海棠MhWRKY40b在几种胁迫下的表达分析[J].园艺学报,2013,40(1):1-9.
[23]	张娜,赵佩,沈法富.陆地棉三个WRKY基因的克隆及表达分析[J].分子植物育种,2012,10(2):169-173.
[24]	Ling J,Jiang W J,Zhang Y,et al. Genome-wide analysis of WRKY gene family in Cucumis sativus[J]. BMC Genomics,2011,12:471.
[25]	Marcel C,John F,Huub J M. WRKY transcription factors involved in activation of SA biosynthesis genes [J]. BMC Plant Biology,2011,11:89.
[26]	Babitha K C,Ramu S V,Pruthvi V,et al. Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress in Arabidopsis[J]. Transgenic Res,2013,22:327-341.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

1. State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Silviculture of State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Received Date: 2014-12-03

Accepted Date: 2015-11-24

Abstract: [Objective]To analyze the sequence features and expression rules of WRKY gene from Corylus heterophylla Fisch. [Method]The gene was cloned by RACE-PCR. Quantitative real-time PCR was used in analyzing gene expression in various tissues and different abiotic stresses, including cold, high salinity and drought. [Result]The cDNA of WRKY is 1 342 bp in length, including a complete open reading frame (ORF) of 963 bp encoding a protein of 320 amino acids, designated as ChWRKY28. Phylogeny tree results showed ChWRKY28 was much closer to AtWRKY28 from Arabidopsis thaliana and PtrWRKY93 from Populus trichocarpa, generated 49% and 60% amino acids similarity. Spatial expression analyses demonstrated that the expression level of ChWRKY28 was higher in stem than that in male anthotaxy and floral buds which indicating tissue-specific expression. ChWRKY28 was clearly induced by cold, high salinity and drought, but that the expression tendency were evidently different of this gene. The subcellular localization analysis showd that ChWRKY28 protein was targeted to the nucleus. [Conclusion]This study indicated that ChWRKY28 gene may be involved in response to abiotic stress signal transduction pathway.

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Reference (26)

[1]	马庆华,王贵禧,梁维坚,等.我国榛属植物种质资源的研究、利用与创新[J]. 果树学报,2013,30(1):159-164.
[2]	Mehlenbacher S A. Hazelnuts. Genetic resources in temperate fruit and nut crops[J]. Acta Horticulturae,1991,290:791-838.
[3]	Liu J,Cheng Y,Liu C,et al. Temporal changes of disodium fluorescein transport in hazelnut during fruit development stage[J]. Sci Hortic Amsterdam,2013,150:348-353.
[4]	Coyne C J, Mehlenbacher S A, Smith D C. Sources of resistance to Eastern Filbert Blight in hazelnut [J]. Journal of the American Society for Horticultural Science,1998,123(2):253-257.
[5]	吕跃东,董凤祥,王贵禧,等. 平欧杂交榛抗寒性综合评价体系的建立与应用[J]. 林业科学,2008,44(9):31-35.
[6]	彭立新,王明启,梁维坚,等. 榛属(Corylus L.)植物抗寒性研究[J]. 吉林林学院学报,1994,10(3):166-170.
[7]	赵天田,陈新,刘庆忠,等.平榛NAC转录因子的分离及表达特性分析[J]. 植物遗传资源学报,2012,13(2):265-270.
[8]	王磊,高晓清,朱苓华,等.植物WRKY转录因子家族基因抗病相关功能的研究进展[J]. 植物遗传资源学报,2011,12(1):80-85.
[9]	Wang H H,Hao J J,Chen X J,et al. Overexpression of rice WRKY89 enhances ultraviolet B tolerance and disease resistance in rice plants[J]. Plant Molecular Biology,2007,65:799-815.
[10]	Ulker B,Somssich I E. WRKY transcription factors:from DNA binding towards biological function[J]. Current Opinion in Plant Biology,2004,7(5):491-498.
[11]	Dong J,Chen C,Chen Z. Expression profiles of the Arabidopsis WRKY gene superfamily during plant defense response [J]. Plant Molecular Biology,2003,51 (1):21-37.
[12]	Eulgem T,Somssich I E. Networks of WRKY transcription factors in defense signaling [J]. Current Opinion in Plant Biology,2007,10:366-371.
[13]	Ishiguro S,Nakamura K. Characterization of a cDNA encoding a novel DNA-binding protein,SPF1,that recognizes SP8 sequences in the 50 upstream regions of genes coding for sporamin and beta-amylase from sweet potato [J]. Mol Gen Genet,1994,244(6):563-571.
[14]	Seki M,Narusaka M,Ishida J,et al. Monitoring the expression profiles of 7000 Arabidopsis genes under drought,cold and high-salinity stresses using a full-length cDNA microarray[J]. The Plant Journal,2002,31 (3):279-292.
[15]	Jiang Y,Deyholos M. Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses [J]. Plant Molecular Biology,2009,69 (1-2):91-105.
[16]	Chen X,Zhang J,Liu Q Z,et al. Transcriptome Sequencing and Identification of Cold Tolerance Genes in Hardy Corylus Species (C. heterophylla Fisch) Floral Buds[J]. PLoS One,2013,9(9):e108604.
[17]	Wang L Q,Wang C,Wang D Y,et al. Molecular characterization and transcript profiling of NAC genes in response to abiotic stress in Tamarix hispida[J]. Tree Genetics & Genomes,2014,10(1):157-171.
[18]	Chang S,Puryear J,Cairney J. A simple and eficient method for isolating RNA from pine trees [J]. Plant Molecular Biol Reporter,1993,11(2):113-116.
[19]	Zhang Y,Feng J C. Identification and characterization of the grape WRKY family[J]. Biomed Research International,2014,787680.
[20]	Wang C,Deng P Y,Chen L L,et al. A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco[J]. PLoS One,2013,8(6):e65120.
[21]	Jiang Y Z,Duan Y J,Yin J,et al. Genome-wide identification and characterization of the Populus WRKY transcription factor family and analysis of their expression in response to biotic and abiotic stresses[J]. J Exp Bot,2014,65:6629-6644.
[22]	罗昌国,渠慎春,张计育,等.湖北海棠MhWRKY40b在几种胁迫下的表达分析[J].园艺学报,2013,40(1):1-9.
[23]	张娜,赵佩,沈法富.陆地棉三个WRKY基因的克隆及表达分析[J].分子植物育种,2012,10(2):169-173.
[24]	Ling J,Jiang W J,Zhang Y,et al. Genome-wide analysis of WRKY gene family in Cucumis sativus[J]. BMC Genomics,2011,12:471.
[25]	Marcel C,John F,Huub J M. WRKY transcription factors involved in activation of SA biosynthesis genes [J]. BMC Plant Biology,2011,11:89.
[26]	Babitha K C,Ramu S V,Pruthvi V,et al. Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress in Arabidopsis[J]. Transgenic Res,2013,22:327-341.

Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

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通讯作者: 陈斌, bchen63@163.com

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Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

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