慈竹BeCesA1和BeCesA7基因的克隆及组织表达分析
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摘要
植物的纤维素合成酶(CesA)是纤维素合成途径的关键酶,不仅与纤维素含量相关,也与植物生长发育相关。但有关慈竹CesA基因的克隆与分析,以及GA3对慈竹CesA基因表达的影响未见报道。本研究通过慈竹转录组数据库、在线NCBI基因组数据库及毛竹基因组数据库,同源克隆得到两个CesA基因,BeCesA1(GenBank注册号:KY435488)和BeCesA7 (GenBank注册号:KY435489),分别编码1 078和1 053个氨基酸残基。保守结构域分析及多重序列比对分析表明,慈竹BeCesA1和BeCesA7蛋白均在N端存在一个RING finger结构和两个高度可变区域。组织表达结果表明,BeCesA1和BeCesA7两个基因在竹不同组织中表达模式各不相同,但均在100 cm笋等组织中高表达,然而BeCesA7的表达量明显高于BeCesA1。在竹的发育过程中,BeCesA1基因随着笋的发育表达量逐渐降低,而BeCesA7则随着笋的发育逐渐增高。外源施加GA3后,以BeCesA1的CK未展开叶为参照,BeCesA1在茎秆中的表达量降低,而BeCesA7则在茎秆中的表达量增加。
Cellulose synthase(CesA) plays a key role in cellulose biosynthesis, which is not only related to cellulose content, but also involved in plant growth and development. However, the cloning and analysis of CesA gene from Bambusa emeiensis as well as the effect of GA3 on the expression of CesA gene in Bambusa emeiensis have not been reported. In this paper, two CesA genes, BeCesA1(GenBank accession number: KY435488) and BeCes A7(GenBank accession number: KY435489), which encoding 1 078 and 1 053 amino acid residues, respectively,were obtained based on GenBank transcriptome database, online NCBI genomic database and Phytomania genomic database. Conserved domain analysis and multiple sequence alignment analysis indicated that both the BeCesA1 and BeCesA7 proteins had a RING finger structure and two highly variable regions at the N-terminus. Tissue expression analysis showed that the expression patterns of BeCesA1 and BeCesA7 genes differed from each other in various bamboo tissues. Whereas, BeCesA1 and BeCesA7 both expressed highly in the 100 cm bamboo shoot,while the expression of BeCesA7 was higher than BeCesA1. During bamboo development, the expression of BeCes A1 gene gradually decreased with the development of shoots, which was on the contrary with that of BeCesA7.After the exogenous application of GA3, the expression of BeCesA1 in the stalks decreased while that of BeCesA7 increased in the stalks with the unexpanded leaf of BeCesA1 as reference.
Cellulose synthase(CesA) plays a key role in cellulose biosynthesis, which is not only related to cellulose content, but also involved in plant growth and development. However, the cloning and analysis of CesA gene from Bambusa emeiensis as well as the effect of GA3 on the expression of CesA gene in Bambusa emeiensis have not been reported. In this paper, two CesA genes, BeCesA1(GenBank accession number: KY435488) and BeCes A7(GenBank accession number: KY435489), which encoding 1 078 and 1 053 amino acid residues, respectively,were obtained based on GenBank transcriptome database, online NCBI genomic database and Phytomania genomic database. Conserved domain analysis and multiple sequence alignment analysis indicated that both the BeCesA1 and BeCesA7 proteins had a RING finger structure and two highly variable regions at the N-terminus. Tissue expression analysis showed that the expression patterns of BeCesA1 and BeCesA7 genes differed from each other in various bamboo tissues. Whereas, BeCesA1 and BeCesA7 both expressed highly in the 100 cm bamboo shoot,while the expression of BeCesA7 was higher than BeCesA1. During bamboo development, the expression of BeCes A1 gene gradually decreased with the development of shoots, which was on the contrary with that of BeCesA7.After the exogenous application of GA3, the expression of BeCesA1 in the stalks decreased while that of BeCesA7 increased in the stalks with the unexpanded leaf of BeCesA1 as reference.
引文
Appenzeller L.,Doblin M.,Barreiro R.,Wang H.Y.,Niu X.M.,Kollipara K.,Carrigan L.,Tomes D.,Chapman M.,and Dhugga K.S.,2004,Cellulose synthesis in maize:isolation and expression analysis of the cellulose synthase(Cesa)gene family,Cellulose,11(3-4):287-299
Burn J.E.,Hocart C.H.,Birch R.J.,Cork A.C.,and Williamson R.E.,2002,Functional analysis of the cellulose synthase genes CesA1,CesA2,and CesA3 in Arabidopsis,Plant Physiol.,129(2):797-807
Carpita N.C.,2011,Update on mechanisms of plant cell wall biosynthesis:how plants make cellulose and other(1-4)-β-D-Glycans,Plant Physiol.,155(1):171-184
Cheminant S.,Wild M.,Bouvier F.,Pelletier S.,Renou J.P.,Erhardt M.,Hayes S.,Terry M.J.,Genschik P.,and Achard P.,2011,Dellas regulate chlorophyll and carotenoid biosynthesis to prevent photooxidative damage during seedling deetiolation in Arabidopsis,Plant Cell,23(5):1849-1860
Dai F.,Hill D.A.,and Matsumura S.,2012,Maternity roosts and behaviour of the ussurian tube-nosed bat Murina ussuriensis,Acta Chiropterologica,14(1):93-104
Deng X.B.,Hu S.L.,Cao Y.,Lu X.Q.,Ren P.,and Duan N.,2011,Bioinformatics analysis of cellulose synthase(CesA)of Bambusa oldhamii and Phyllostachy edulis,Fujian Linxueyuan Xuebao(Journal of Fujian College of Forestry),31(1):84-90(邓小波,胡尚连,曹颖,卢学琴,任鹏,段宁,2011,绿竹与毛竹纤维素合成酶(CesA)的生物信息学分析,福建林学院学报,31(1):84-90)
Doblin M.S.,Kurek I.,Jacob-Wilk D.,and Delmer D.P.,2002,Cellulose biosynthesis in plants:from genes to rosettes,Plant Cell Physiol.,43(12):1407-1420
Hai G.,Jia Z.,Xu W.,Wang C.,Cao S.,Liu J.,and Cheng Y.,2016,Characterization of the Populus Ptrcesa4,promoter in transgenic Populus alba×P.glandulosa,Plant Cell Tiss.Org.,124(3):495-505
Huang D.B.,Wang S.G.,Zhang B.C.,Shang-Guan K.,Shi Y.Y.,Zhang D.M.,Liu X.L.,Wu K.,Xu Z.P.,Fu X.D.,and Zhou Y.H.,2015,A gibberellin-mediated della-nac signaling cascade regulates cellulose synthesis in rice,Plant Cell,27(6):1681-1696
Ji L.,and Yang R.C.,2002,Rice stem elongation and plant hormones,Zhiwuxue Tongbao(Chinese Bulletin of Botany),19(1):109-115(季兰,杨仁崔,2002,水稻茎伸长生长与植物激素,植物学通报,19(1):109-115)
Jiang J.,Jie Y.C.,Zhou Q.M.,Zhou J.H.,Zhu S.J.,Xing H.C.,and Zhong Y.L.,2012,Full-length cDNA cloning and express analysis of BnCesA1 in ramie,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),13(5):851-857(蒋杰,揭雨成,周清明,周精华,朱守晶,邢虎成,钟英丽,2012,苎麻纤维素合酶基因BnCesA1全长cDNA的克隆与表达分析,植物遗传资源学报,13(5):851-857)
Jiang X.,Lou C.,Yuan N.,Yue J.J.,and Gu X.P.,2016,Effect of exogenous GA3on culm form and culm fiber quality of Phyllostachys edulis seedling of neonatal tiller,Nanjing Linye Daxue Xuebao(Journal of Nanjing Forestry University),40(2):121-126(江雪,楼崇,袁娜,岳晋军,顾小平,2016,外源GA3对毛竹实生苗新分蘖竹株秆形与竹材纤维质量的影响,南京林业大学学报,40(2):121-126)
Jiang Y.,Hu S.L.,Cao Y.,Lu X.Q.,Huang Y.,and Xu G.,2017,Effects of exogenous GA3on the growth of stem and CesAgene expression in Phyllostachys edulis seedling,Zhiwu Yanjiu(Bulletin of Botanical Research),37(5):744-750(姜勇,胡尚连,曹颖,卢学琴,黄艳,徐刚,2017,外源GA3对毛竹实生苗茎秆生长及CesA基因表达的影响,植物研究,37(5):744-750)
Li F.L.,Chen J.C.,and Zhao Y.J.,2000,Effect of gibberellin and light on seed germination and seedling growth of Arabidopsis,Zhiwu Shengli Xuebao(Acta Phytophysiologica Sinica),26(2):101-104(李凤玲,陈季楚,赵毓橘,2000,赤霉素和光对拟南芥种子萌发和幼苗生长的影响,植物生理学报,26(2):101-104)
Li S.D,Lei L.,and Gu Y.,2013,Functional analysis of complexes with mixed primary and secondary cellulose synthases,Plant Signal.Behav.,8(3):e23179
McFarlane H.E.,Doring A.,and Persson S.,2014,The cell biology of cellulose synthesis,Annu.Rev.Plant Biol.,65:69-94
Ogawa M.,Hanada A.,Yamauchi Y.,Kuwahara A.,Kamiya Y.,and Yamaguchi S.,2003,Gibberellin biosynthesis and response during Arabidopsis seed germination,Plant Cell,15(7):1591-1604
Somerville C.,2016,Cellulose synthesis in higher plants,Annu.Rev.Cell Dev.Biol.,22(1):53-78
Taylor N.G.,Howells R.M.,Huttly A.K.,Vickers K.,and Turner S.R.,2003,Interactions among three distinct CesA proteins essential for cellulose synthesis,Proc.Natl.Acad.Sci.USA,100(3):1450-1455
Tian Z.J.,Yi R.,Chen J.R.,Guo Q.Q.,and Zhang X.W.,2008,Cloning and expression of cellulose synthase gene in ramie(Boehmeria nivea(Linn.)Gaud.),Zuowu Xuebao(Acta A-gronomica Sinica),34(1):76-83(田志坚,易蓉,陈建荣,郭清泉,张学文,2008,苎麻纤维素合成酶基因cDNA的克隆及表达分析,作物学报,34(1):76-83)
Turner S.R.,and Somerville C.R.,1997,Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall,Plant Cell,9(5):689-701
Wang L.Q.,Guo K.,Li Y.,Tu Y.Y.,Hu H.Z.,Wang B.R.,Cui X.C.,and Peng L.C.,2010,Expression profiling and integrative analysis of the CESA/CSL superfamily in rice,BMCPlant Biol.,10(1):282
Yuan L.C.,2009,Study on the gene involved in the cellulose biosynthesis of moso bamboo(Phyllostachys edulis),Dissertation for Ph.D.,Chinese Academy of Forestry,Supercvisor:Peng Z.H.,pp.31-52(袁丽钗,2009,毛竹纤维素生物合成相关基因研究,博士学位论文,中国林业科学研究院,导师:彭镇华,pp.31-52)
Burn J.E.,Hocart C.H.,Birch R.J.,Cork A.C.,and Williamson R.E.,2002,Functional analysis of the cellulose synthase genes CesA1,CesA2,and CesA3 in Arabidopsis,Plant Physiol.,129(2):797-807
Carpita N.C.,2011,Update on mechanisms of plant cell wall biosynthesis:how plants make cellulose and other(1-4)-β-D-Glycans,Plant Physiol.,155(1):171-184
Cheminant S.,Wild M.,Bouvier F.,Pelletier S.,Renou J.P.,Erhardt M.,Hayes S.,Terry M.J.,Genschik P.,and Achard P.,2011,Dellas regulate chlorophyll and carotenoid biosynthesis to prevent photooxidative damage during seedling deetiolation in Arabidopsis,Plant Cell,23(5):1849-1860
Dai F.,Hill D.A.,and Matsumura S.,2012,Maternity roosts and behaviour of the ussurian tube-nosed bat Murina ussuriensis,Acta Chiropterologica,14(1):93-104
Deng X.B.,Hu S.L.,Cao Y.,Lu X.Q.,Ren P.,and Duan N.,2011,Bioinformatics analysis of cellulose synthase(CesA)of Bambusa oldhamii and Phyllostachy edulis,Fujian Linxueyuan Xuebao(Journal of Fujian College of Forestry),31(1):84-90(邓小波,胡尚连,曹颖,卢学琴,任鹏,段宁,2011,绿竹与毛竹纤维素合成酶(CesA)的生物信息学分析,福建林学院学报,31(1):84-90)
Doblin M.S.,Kurek I.,Jacob-Wilk D.,and Delmer D.P.,2002,Cellulose biosynthesis in plants:from genes to rosettes,Plant Cell Physiol.,43(12):1407-1420
Hai G.,Jia Z.,Xu W.,Wang C.,Cao S.,Liu J.,and Cheng Y.,2016,Characterization of the Populus Ptrcesa4,promoter in transgenic Populus alba×P.glandulosa,Plant Cell Tiss.Org.,124(3):495-505
Huang D.B.,Wang S.G.,Zhang B.C.,Shang-Guan K.,Shi Y.Y.,Zhang D.M.,Liu X.L.,Wu K.,Xu Z.P.,Fu X.D.,and Zhou Y.H.,2015,A gibberellin-mediated della-nac signaling cascade regulates cellulose synthesis in rice,Plant Cell,27(6):1681-1696
Ji L.,and Yang R.C.,2002,Rice stem elongation and plant hormones,Zhiwuxue Tongbao(Chinese Bulletin of Botany),19(1):109-115(季兰,杨仁崔,2002,水稻茎伸长生长与植物激素,植物学通报,19(1):109-115)
Jiang J.,Jie Y.C.,Zhou Q.M.,Zhou J.H.,Zhu S.J.,Xing H.C.,and Zhong Y.L.,2012,Full-length cDNA cloning and express analysis of BnCesA1 in ramie,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),13(5):851-857(蒋杰,揭雨成,周清明,周精华,朱守晶,邢虎成,钟英丽,2012,苎麻纤维素合酶基因BnCesA1全长cDNA的克隆与表达分析,植物遗传资源学报,13(5):851-857)
Jiang X.,Lou C.,Yuan N.,Yue J.J.,and Gu X.P.,2016,Effect of exogenous GA3on culm form and culm fiber quality of Phyllostachys edulis seedling of neonatal tiller,Nanjing Linye Daxue Xuebao(Journal of Nanjing Forestry University),40(2):121-126(江雪,楼崇,袁娜,岳晋军,顾小平,2016,外源GA3对毛竹实生苗新分蘖竹株秆形与竹材纤维质量的影响,南京林业大学学报,40(2):121-126)
Jiang Y.,Hu S.L.,Cao Y.,Lu X.Q.,Huang Y.,and Xu G.,2017,Effects of exogenous GA3on the growth of stem and CesAgene expression in Phyllostachys edulis seedling,Zhiwu Yanjiu(Bulletin of Botanical Research),37(5):744-750(姜勇,胡尚连,曹颖,卢学琴,黄艳,徐刚,2017,外源GA3对毛竹实生苗茎秆生长及CesA基因表达的影响,植物研究,37(5):744-750)
Li F.L.,Chen J.C.,and Zhao Y.J.,2000,Effect of gibberellin and light on seed germination and seedling growth of Arabidopsis,Zhiwu Shengli Xuebao(Acta Phytophysiologica Sinica),26(2):101-104(李凤玲,陈季楚,赵毓橘,2000,赤霉素和光对拟南芥种子萌发和幼苗生长的影响,植物生理学报,26(2):101-104)
Li S.D,Lei L.,and Gu Y.,2013,Functional analysis of complexes with mixed primary and secondary cellulose synthases,Plant Signal.Behav.,8(3):e23179
McFarlane H.E.,Doring A.,and Persson S.,2014,The cell biology of cellulose synthesis,Annu.Rev.Plant Biol.,65:69-94
Ogawa M.,Hanada A.,Yamauchi Y.,Kuwahara A.,Kamiya Y.,and Yamaguchi S.,2003,Gibberellin biosynthesis and response during Arabidopsis seed germination,Plant Cell,15(7):1591-1604
Somerville C.,2016,Cellulose synthesis in higher plants,Annu.Rev.Cell Dev.Biol.,22(1):53-78
Taylor N.G.,Howells R.M.,Huttly A.K.,Vickers K.,and Turner S.R.,2003,Interactions among three distinct CesA proteins essential for cellulose synthesis,Proc.Natl.Acad.Sci.USA,100(3):1450-1455
Tian Z.J.,Yi R.,Chen J.R.,Guo Q.Q.,and Zhang X.W.,2008,Cloning and expression of cellulose synthase gene in ramie(Boehmeria nivea(Linn.)Gaud.),Zuowu Xuebao(Acta A-gronomica Sinica),34(1):76-83(田志坚,易蓉,陈建荣,郭清泉,张学文,2008,苎麻纤维素合成酶基因cDNA的克隆及表达分析,作物学报,34(1):76-83)
Turner S.R.,and Somerville C.R.,1997,Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall,Plant Cell,9(5):689-701
Wang L.Q.,Guo K.,Li Y.,Tu Y.Y.,Hu H.Z.,Wang B.R.,Cui X.C.,and Peng L.C.,2010,Expression profiling and integrative analysis of the CESA/CSL superfamily in rice,BMCPlant Biol.,10(1):282
Yuan L.C.,2009,Study on the gene involved in the cellulose biosynthesis of moso bamboo(Phyllostachys edulis),Dissertation for Ph.D.,Chinese Academy of Forestry,Supercvisor:Peng Z.H.,pp.31-52(袁丽钗,2009,毛竹纤维素生物合成相关基因研究,博士学位论文,中国林业科学研究院,导师:彭镇华,pp.31-52)
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