沙柳SpsTAC2基因克隆、生物信息学及组织特异表达分析
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摘要
TAC基因隶属IGT基因家族,该基因家族主要参与植物分枝角度的调控,目前国内外针对木本植物中TAC基因的研究较少。为了获得沙柳中的TAC基因并对该序列进行初步分析,本研究用同源克隆技术在沙柳中克隆得到长度为783 bp的CDS序列并命名为Sps TAC2基因,对其进行生物信息学及组织特异性表达分析。结果分析表明,该基因编码260个氨基酸,为亲水性蛋白,未发现跨膜结构域,无信号肽区域,亚细胞定位预测在细胞核,基因内含有IGT基因家族典型结构域。系统进化树将SpsTAC2氨基酸序列与杨、柳位于第二条染色体上的TAC-like基因所编码氨基酸序列聚为一类。RT-qPCR结果表明,该基因在成熟叶表达量最高,其次是韧皮部,在根部几乎不表达。从沙柳中成功克隆出了调控分枝角度的IGT基因家族的SpsTAC2基因,推测该基因的主要作用部位可能在成熟叶、茎中。沙柳SpsTAC2基因的研究为IGT基因家族研究及沙柳育种工作提供了理论依据。
TAC gene belongs to IGT gene family which is mainly involved in the regulation of plant branching angle. At present, there are few studies on TAC genes in xylophyta at home and abroad. In order to obtain the TAC gene from Salix psammophila and to preliminarily analyze the sequence, a 783 bp CDS sequence was cloned from Salix psammophila by homologous cloning method in this study, which was named as SpsTAC2 gene, and then bioinformatics and tissue specific expression analysis of this gene was performed. The result showed that SpsTAC2 gene encoded 260 amino acid, which was hydrophilic proteins without transmembrane structure and signal peptide. It was located in nucleus by subcellular localization prediction, and it contained typically conserved motifs of IGT gene family. Phylogenetic tree classified SpsTAC2 amino acid sequence with TAC-like gene coding amino acid sequence of poplar and willow on the second chromosome. The RT-qPCR results indicated that SpsTAC2 gene had the highest expression in the mature leaves, followed by phloems, and the expression was the lowest in roots. In this study, the SpsTAC2 gene in IGT gene family which could regulate branch angle was successfully cloned from Salix psammophila. It was speculated that this gene might have important function in mature leaves and stems. The research of SpsTAC2 gene in Salix psammophila could provide theoretical basis for IGT gene family exploration and breeding of Salix psammophila.
TAC gene belongs to IGT gene family which is mainly involved in the regulation of plant branching angle. At present, there are few studies on TAC genes in xylophyta at home and abroad. In order to obtain the TAC gene from Salix psammophila and to preliminarily analyze the sequence, a 783 bp CDS sequence was cloned from Salix psammophila by homologous cloning method in this study, which was named as SpsTAC2 gene, and then bioinformatics and tissue specific expression analysis of this gene was performed. The result showed that SpsTAC2 gene encoded 260 amino acid, which was hydrophilic proteins without transmembrane structure and signal peptide. It was located in nucleus by subcellular localization prediction, and it contained typically conserved motifs of IGT gene family. Phylogenetic tree classified SpsTAC2 amino acid sequence with TAC-like gene coding amino acid sequence of poplar and willow on the second chromosome. The RT-qPCR results indicated that SpsTAC2 gene had the highest expression in the mature leaves, followed by phloems, and the expression was the lowest in roots. In this study, the SpsTAC2 gene in IGT gene family which could regulate branch angle was successfully cloned from Salix psammophila. It was speculated that this gene might have important function in mature leaves and stems. The research of SpsTAC2 gene in Salix psammophila could provide theoretical basis for IGT gene family exploration and breeding of Salix psammophila.
引文
Cao X.,Deng M.,Zhang Z.L.,Liu Y.J.,Yang X.L.,Zhou H.,and Liu Y.X.,2017,Molecular characterization and expression analysis of TaTAC1 gene in Triticum aestivum L.,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),18(1):125-132(曹鑫,邓梅,张正丽,刘宇娇,杨希兰,周红,刘亚西,2017,小麦分蘖角度TaTAC1基因同源克隆及表达分析,植物遗传资源学报,18(1):125-132)
Chen Z.X.,Zuo S.M.,Zhang Y.F.,Chen H.Q.,Feng M.F.,Jiang W.,Feng F.,Ma Y.Y.,Hua H.L.,Li G.S.,and Pan X.B.,2014,Breeding potential and interaction effects of qSB-9TQand TAC1TQin rice breeding against sheath blight disease,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),28(5):479-486(陈宗祥,左示敏,张亚芳,陈红旗,冯明辉,姜伟,冯凡,马玉银,华鹤良,李国生,潘学彪,2014,抗纹枯病基因q SB-9TQ和分蘖角基因TAC1TQ在抗病育种中的互作效应及育种价值,中国水稻科学,28(5):479-486)
Dardick C.,Callahan A.,Horn R.,Ruiz K.,Zhebentyayeva T.,Hollender C.,Whitaker M.,Abbott A.,and Scorza R.,2013,PpeTAC1 promotes the horizontal growth of branches in peach trees and is a member of a functionally conserved gene family found in diverse plants species,Plant J.,75(4):618-630
Dong Z.,Jiang C.,Chen X.Y.,Zhang T.,Ding L.,Song W.B.,Luo H.B.,Lai J.S.,Chen H.B.,Liu R.Y.,Zhang X.L.,and Jin W.W.,2013,Maize LAZY1 mediates shoot gravitropism and inflorescence development through regulating auxin transport,auxin signaling,and light response,Plant Physiol.,163(3):1306-1322
Guseman J.M.,Webb K.,Srinivasan C.,and Dardick C.,2017,DRO1 influences root system architecture in Arabidopsis and Prunus species,Plant J.,89(6):1093-1105
Hollender C.A.,and Dardick C.,2015,Molecular basis of angiosperm tree architecture,New Phytologist,206(2):541-556
Hou J.,2016,Genomic bases leading to the divergence between willow and poplar and the differentiation in their sex determination systems,Dissertation for Ph.D.,Nanjing Forestry University,Supervisor:Yin T.M.,pp.20-25(侯静,2016,杨柳分化及其性别决定的基因组学基础,博士学位论文,南京林业大学,导师:尹佟明,pp.20-25)
Jiang J.H.,Tan L.B.,Zhu Z.F.,Fu Y.C.,Liu F.X.,Cai H.W.,and Sun C.Q.,2012,Molecular evolution of the TAC1 gene from rice(Oryza sativa L.),J.Genet.Genomics,39(10):551-560
Ku L.X.,Wei X.M.,Zhang S.F.,Zhang J.,Guo S.L.,and Chen Y.H.,2011,Cloning and characterization of a putative TAC1ortholog associated with leaf angle in maize(Zea mays L.),Plo S One,6(6):e20621
Li P.,Wang Y.H.,Qian Q.,Fu Z.M.,Wang M.,Zeng D.,Li B.H.,Wang X.J.,and Li J.Y.,2007,LAZY1 controls rice shoot gravitropism through regulating polar auxin transport,Cell Res.,17(5):402-410
Li X.,Huang Y.M.,Gong J.R.,and Zhang X.S.,2010,A study of the development of bio-energy resources and the status of eco-society in China,Energy,35(11):4451-4456
Ma Y.L.,2016,Cloning and bioinformatics of TAC1 in Salix psammophila and characterization of its tissue expression,Inner Mongolia Agricultural University,Supervisor:Yang H.F.,pp.20-25(马月林,2016,沙柳TAC1基因克隆与生物信息学及组织表达特性分析,硕士学位论文,内蒙古农业大学,导师:杨海峰,pp.20-25)
Qi X.,2015,Research on branch angle genes PopTAC and PopLAXY in narrow crown poplar,Shandong Agricultural University,Supervisor:Li J.H.,pp.23(亓晓,2015,窄冠型杨树分枝相关基因PopTAC、PopLAXY的克隆及功能分析,硕士学位论文,山东农业大学,导师:李际红,pp.23)
Surigega,Wang T.J.,Sun H.Y.,Yuan Y.L.,and Du J.R.,2015,Studies on architectural characteristics and sand-fixing effects of three Artemisia plants in Horqin sandy land,Neimenggu Daxue Xuebao(Journal of Inner Mongolia A-gricultural University(Natural Science Edition)),46(3):270-276(苏日格嘎,王铁娟,孙海玉,员艳丽,杜俊瑞,2015,科尔沁沙地三种沙蒿植株构型特征与防风固沙效应研究,内蒙古大学学报(自然科学版),46(3):270-276)
Tuskan G.A.,Difazio S.,Faivre-Rampant P.,Gaudet M.,Harfouche A.,Jorge V.,LabbéJ.L.,Ranjan P.,Sabatti M.,Slavov G.,Street N.,Tschaplinski T.J.,and Yinet T.,2012,The obscure events contributing to the evolution of an incipient sex chromosome in Populus:a retrospective working hypothesis,Tree Genet.Genomes,8(3):559-571
Uga Y.,Okuno K.,and Yano M.,2011,Dro1,a major QTL involved in deep rooting of rice under upland field conditions,J.Exp.Bot.,62(8):2485-2494
Uga Y.,Sugimoto K.,Ogawa S.,Rane J.,Ishitani M.,Hara N.,Kitomi Y.,Inukai Y.,Ono K.,Kanno N.,Inoue H.,Takehisa H.,Motoyama R.,Nagamura Y.,Wu J.Z.,Matsumoto T.,Takai T.,Okuno K.,and Yano M.,2013,Control of root system architecture by DEEPER ROOTING 1 increases rice yield under drought conditions,Nat.Genet.,45(9):1097-1102
Wen M.X.,Chen A.D.,Bian N.F.,Yang H.F.,Qu C.X.,and Wang X.E.,2011,Research on genes related to development of plant shoot branch,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),23(4):111-114(温明星,陈爱大,卞能飞,杨红福,曲朝喜,王秀娥,2011,植物分枝发育的相关基因研究,江西农业学报,23(4):111-114)
Yoshihara T.,and Iino M.,2007,Identification of the gravitropism-related rice gene LAZY1 and elucidation of LAZY1-dependent and-independent gravity signaling pathways,Plant Cell Physiol.,48(5):678-688
Yu B.,Lin Z.W.,Li H.X.,Li X.J.,Li J.Y.,Wang Y.H.,Zhang X.,Zhu Z.F.,Zhai W.X.,Wang X.K.,Xie D.X.,and Sun C.Q.,2007,TAC1,a major quantitative trait locus controlling tiller angle in rice,Plant J.,52(5):891-898
Zhang L.,Ye Z.W.,Zhao C.X.,Zhang G.S.,and Yang H.F.,2017,Cloning and bioinformatics analysis of LAZY1a and LAZY1b genes in Salix psammophila,Xibei Linxueyuan Xuebao(Journal of Northwest Forestry University),32(1):98-105(张磊,叶志玮,赵晨曦,张国盛,杨海峰,2017,沙柳SpsLAZY1a和SpsLAZY1b基因克隆及生物信息学分析,西北林学院学报,32(1):98-105)
Zhao H.,Huai Z.X.,Xiao Y.J.,Wang X.H.,Yu J.Y.,Ding G.D.,and Peng J.H.,2014,Natural variation and genetic analysis of the tiller angle gene MsTAC1 in Miscanthus sinensis,Planta,240(1):161-175
Chen Z.X.,Zuo S.M.,Zhang Y.F.,Chen H.Q.,Feng M.F.,Jiang W.,Feng F.,Ma Y.Y.,Hua H.L.,Li G.S.,and Pan X.B.,2014,Breeding potential and interaction effects of qSB-9TQand TAC1TQin rice breeding against sheath blight disease,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),28(5):479-486(陈宗祥,左示敏,张亚芳,陈红旗,冯明辉,姜伟,冯凡,马玉银,华鹤良,李国生,潘学彪,2014,抗纹枯病基因q SB-9TQ和分蘖角基因TAC1TQ在抗病育种中的互作效应及育种价值,中国水稻科学,28(5):479-486)
Dardick C.,Callahan A.,Horn R.,Ruiz K.,Zhebentyayeva T.,Hollender C.,Whitaker M.,Abbott A.,and Scorza R.,2013,PpeTAC1 promotes the horizontal growth of branches in peach trees and is a member of a functionally conserved gene family found in diverse plants species,Plant J.,75(4):618-630
Dong Z.,Jiang C.,Chen X.Y.,Zhang T.,Ding L.,Song W.B.,Luo H.B.,Lai J.S.,Chen H.B.,Liu R.Y.,Zhang X.L.,and Jin W.W.,2013,Maize LAZY1 mediates shoot gravitropism and inflorescence development through regulating auxin transport,auxin signaling,and light response,Plant Physiol.,163(3):1306-1322
Guseman J.M.,Webb K.,Srinivasan C.,and Dardick C.,2017,DRO1 influences root system architecture in Arabidopsis and Prunus species,Plant J.,89(6):1093-1105
Hollender C.A.,and Dardick C.,2015,Molecular basis of angiosperm tree architecture,New Phytologist,206(2):541-556
Hou J.,2016,Genomic bases leading to the divergence between willow and poplar and the differentiation in their sex determination systems,Dissertation for Ph.D.,Nanjing Forestry University,Supervisor:Yin T.M.,pp.20-25(侯静,2016,杨柳分化及其性别决定的基因组学基础,博士学位论文,南京林业大学,导师:尹佟明,pp.20-25)
Jiang J.H.,Tan L.B.,Zhu Z.F.,Fu Y.C.,Liu F.X.,Cai H.W.,and Sun C.Q.,2012,Molecular evolution of the TAC1 gene from rice(Oryza sativa L.),J.Genet.Genomics,39(10):551-560
Ku L.X.,Wei X.M.,Zhang S.F.,Zhang J.,Guo S.L.,and Chen Y.H.,2011,Cloning and characterization of a putative TAC1ortholog associated with leaf angle in maize(Zea mays L.),Plo S One,6(6):e20621
Li P.,Wang Y.H.,Qian Q.,Fu Z.M.,Wang M.,Zeng D.,Li B.H.,Wang X.J.,and Li J.Y.,2007,LAZY1 controls rice shoot gravitropism through regulating polar auxin transport,Cell Res.,17(5):402-410
Li X.,Huang Y.M.,Gong J.R.,and Zhang X.S.,2010,A study of the development of bio-energy resources and the status of eco-society in China,Energy,35(11):4451-4456
Ma Y.L.,2016,Cloning and bioinformatics of TAC1 in Salix psammophila and characterization of its tissue expression,Inner Mongolia Agricultural University,Supervisor:Yang H.F.,pp.20-25(马月林,2016,沙柳TAC1基因克隆与生物信息学及组织表达特性分析,硕士学位论文,内蒙古农业大学,导师:杨海峰,pp.20-25)
Qi X.,2015,Research on branch angle genes PopTAC and PopLAXY in narrow crown poplar,Shandong Agricultural University,Supervisor:Li J.H.,pp.23(亓晓,2015,窄冠型杨树分枝相关基因PopTAC、PopLAXY的克隆及功能分析,硕士学位论文,山东农业大学,导师:李际红,pp.23)
Surigega,Wang T.J.,Sun H.Y.,Yuan Y.L.,and Du J.R.,2015,Studies on architectural characteristics and sand-fixing effects of three Artemisia plants in Horqin sandy land,Neimenggu Daxue Xuebao(Journal of Inner Mongolia A-gricultural University(Natural Science Edition)),46(3):270-276(苏日格嘎,王铁娟,孙海玉,员艳丽,杜俊瑞,2015,科尔沁沙地三种沙蒿植株构型特征与防风固沙效应研究,内蒙古大学学报(自然科学版),46(3):270-276)
Tuskan G.A.,Difazio S.,Faivre-Rampant P.,Gaudet M.,Harfouche A.,Jorge V.,LabbéJ.L.,Ranjan P.,Sabatti M.,Slavov G.,Street N.,Tschaplinski T.J.,and Yinet T.,2012,The obscure events contributing to the evolution of an incipient sex chromosome in Populus:a retrospective working hypothesis,Tree Genet.Genomes,8(3):559-571
Uga Y.,Okuno K.,and Yano M.,2011,Dro1,a major QTL involved in deep rooting of rice under upland field conditions,J.Exp.Bot.,62(8):2485-2494
Uga Y.,Sugimoto K.,Ogawa S.,Rane J.,Ishitani M.,Hara N.,Kitomi Y.,Inukai Y.,Ono K.,Kanno N.,Inoue H.,Takehisa H.,Motoyama R.,Nagamura Y.,Wu J.Z.,Matsumoto T.,Takai T.,Okuno K.,and Yano M.,2013,Control of root system architecture by DEEPER ROOTING 1 increases rice yield under drought conditions,Nat.Genet.,45(9):1097-1102
Wen M.X.,Chen A.D.,Bian N.F.,Yang H.F.,Qu C.X.,and Wang X.E.,2011,Research on genes related to development of plant shoot branch,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),23(4):111-114(温明星,陈爱大,卞能飞,杨红福,曲朝喜,王秀娥,2011,植物分枝发育的相关基因研究,江西农业学报,23(4):111-114)
Yoshihara T.,and Iino M.,2007,Identification of the gravitropism-related rice gene LAZY1 and elucidation of LAZY1-dependent and-independent gravity signaling pathways,Plant Cell Physiol.,48(5):678-688
Yu B.,Lin Z.W.,Li H.X.,Li X.J.,Li J.Y.,Wang Y.H.,Zhang X.,Zhu Z.F.,Zhai W.X.,Wang X.K.,Xie D.X.,and Sun C.Q.,2007,TAC1,a major quantitative trait locus controlling tiller angle in rice,Plant J.,52(5):891-898
Zhang L.,Ye Z.W.,Zhao C.X.,Zhang G.S.,and Yang H.F.,2017,Cloning and bioinformatics analysis of LAZY1a and LAZY1b genes in Salix psammophila,Xibei Linxueyuan Xuebao(Journal of Northwest Forestry University),32(1):98-105(张磊,叶志玮,赵晨曦,张国盛,杨海峰,2017,沙柳SpsLAZY1a和SpsLAZY1b基因克隆及生物信息学分析,西北林学院学报,32(1):98-105)
Zhao H.,Huai Z.X.,Xiao Y.J.,Wang X.H.,Yu J.Y.,Ding G.D.,and Peng J.H.,2014,Natural variation and genetic analysis of the tiller angle gene MsTAC1 in Miscanthus sinensis,Planta,240(1):161-175