苹果砧木YUCCA10a基因的克隆及其在嫁接树体的定量表达分析
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摘要
为了研究M9矮化砧和八棱海棠砧YUCCA10a基因的结构特征及其嫁接树体的表达特征,从M9矮化砧和八棱海棠砧中克隆了YUCCA10a基因,并对该基因进行了序列分析以及不同时期2种砧木嫁接"烟富3"的定量表达分析。通过分析表明,该基因的编码序列为1 149 bp,推导编码382个氨基酸,预测蛋白质分子量为94 285.29 Da,理论等电点为5.08。实时荧光定量PCR结果表明,5月和10月份的M9矮化砧嫁接"烟富3"中YUCCA10a基因相对表达量明显低于八棱海棠砧,2种砧木嫁接的"烟富3"中YUCCA10a基因在10月份的相对表达量略高于5月份。本试验结果为研究M9矮化砧和八棱海棠砧YUCCA10a基因的结构特征和砧木对嫁接"烟富3"影响的机理方面提供理论依据。
In order to study the structural characteristics and the expression characteristics of YUCCA10 a gene in grafted tree of M9 dwarfing rootstock and Baleng crab rootstock, YUCCA10 a gene was cloned and analysed from M9 dwarfing rootstock and Baleng crab rootstock. Quantitative expression of YUCCA10 a gene from scion in different growth periods of 'Yanfu 3', which was grafted on M9 and Baleng crab rootstocks, was also analysed. The study showed that the coding sequence of the gene was 1 149 bp, deduced to encode 382 amino acids, the predicted molecular weight of the protein was 94 285.29 Da, and the theoretical isoelectric point was 5.08. The results of real-time fluorescence quantitative PCR showed that the relative expression of YUCCA10 a gene was significantly lower in the 'Yanfu 3' grafted with M9 dwarfing rootstock in the scion materials collected in May and in October than that of Baleng crab rootstock. The relative expression of YUCCA10 a gene in 'Yanfu 3' grafted with two kinds of rootstocks in October was slightly higher than that in May. The results of this study could provide a theoretical basis for studying the structural characteristics of the YUCCA10 a gene of the M9 dwarfing rootstock and Baleng crab rootstocks and the mechanism of the rootstock affecting the grafting of 'Yanfu 3'.
In order to study the structural characteristics and the expression characteristics of YUCCA10 a gene in grafted tree of M9 dwarfing rootstock and Baleng crab rootstock, YUCCA10 a gene was cloned and analysed from M9 dwarfing rootstock and Baleng crab rootstock. Quantitative expression of YUCCA10 a gene from scion in different growth periods of 'Yanfu 3', which was grafted on M9 and Baleng crab rootstocks, was also analysed. The study showed that the coding sequence of the gene was 1 149 bp, deduced to encode 382 amino acids, the predicted molecular weight of the protein was 94 285.29 Da, and the theoretical isoelectric point was 5.08. The results of real-time fluorescence quantitative PCR showed that the relative expression of YUCCA10 a gene was significantly lower in the 'Yanfu 3' grafted with M9 dwarfing rootstock in the scion materials collected in May and in October than that of Baleng crab rootstock. The relative expression of YUCCA10 a gene in 'Yanfu 3' grafted with two kinds of rootstocks in October was slightly higher than that in May. The results of this study could provide a theoretical basis for studying the structural characteristics of the YUCCA10 a gene of the M9 dwarfing rootstock and Baleng crab rootstocks and the mechanism of the rootstock affecting the grafting of 'Yanfu 3'.
引文
Cheng Y.F.,Dai X.H.,and Zhao Y.D.,2006,Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis,Genes Dev.,20(13):1790-1799
Cheng Y.F.,Dai X.H.,and Zhao Y.D.,2007,Auxin synthesized by the YUCCA flavin monooxygenases is essential for embryogenesis and leaf formation in Arabidopsis,Plant Cell,19(8):2430-2439
Davies P.J.,ed.,1995,Plant hormones:physiology,biochemistry and molecular biology,Springer Netherlands,Berlin,Germany,pp.1-15
Liu X.Y.,Li J.,Liu M.M.,Yao Q.,and Chen J.Z.,2017,Transcriptome profiling to understand the effect of citrus rootstocks on the growth of'Shatangju'mandarin,PLoS One,12(1):e0169897
Mano Y.,and Nemoto K.,2012,The pathway of auxin biosynthesis in plants,J.Exp.Bot.,63(8):2853-2872
Sambrook J.,and Russell D.W.,eds.,Huang P.T.,trans.,2016,Molecular cloning:a laboratory manuals,Science Press,Beijing,China,pp.865(萨姆布鲁克J.,拉塞尔D.W.,主编,黄培堂,主译,2016,分子克隆实验指南,科学出版社,中国,北京,pp.865)
Santos A.,Ribeiro R.,and CrespíA.L.,2004,Sweet cherry(Prunus avium)growth is mostly affected by rootstock and much less by budding height,New Zeal.J.Crop Hort.,32(3):309-318
Soejima J.,Yoshida Y.,Haniuda T.,Bessho H.,Tsuchiya S.,Masuda T.,Komori S.,Sanada T.,Ito Y.,Sadamori S.,and Kashimura T.,2010,New dwarfing apple rootstocks'JM 1','JM7'and'JM 8',Bull.Natl.Inst.Fruit Tree Sci.,11:1-16
Song C.H.,2017,Study the role of several key IAA meatbolism genes in dwarf rootstock induced scion dwarfing and the role of mi RNA in the spur type apple dwarfing,Dissertation for Ph.D.,Northwest A&F University,Supervisor:Han Y.M.,pp.1-124(宋春晖,2017,苹果IAA代谢几个关键基因在矮化砧木与mi RNA在短枝品种中致矮作用研究,博士学位论文,西北农林科技大学,导师:韩玉明,pp.1-124)
Song C.H.,Zhang D.,Zhang J.,Zheng L.W.,Zhao C.P.,Ma J.J.,An N.,and Han M.Y.,2016,Expression analysis of key auxin synthesis,transport,and metabolism genes in different young dwarfing apple trees,Acta Physiol.Plant.,38(2):1-15
Song J.C,Jiang L.J,and Jameson P.E.,2012,Co-ordinate regulation of cytokinin gene family members during flag leaf and reproductive development in wheat,BMC Plant Biol.,12(1):78-94
Won C.,Shen X.L.,Mashiguchi K.,Zheng Z.Y.,Dai X.H.,Cheng Y.H.,Kasahara H.,Kamiya Y.,Chory J.,and Zhao Y.D.,2011,Conversion of tryptophan to indole-3-acetic acid by TRYPTOPHAN AMINOTRANSFERASES OF ARABIDO-PSIS and YUCCAs in Arabidopsis,Proc.Natl.Acad.Sci.USA,108(45):18518-18523
Zhao Y.,2010,Auxin biosynthesis and its role in plant development,Ann.Rev.Plant Biol.,61(1):49-64
Zhao Y.,Christensen K.S.,Fankhauser C.,Cashman J.R.,Cohen J.D.,Weigel D.,and Chory J.,2001,A role for flavin monooxygenase-like enzymes in auxin biosynthesis,Science,291(5502):306-309
Cheng Y.F.,Dai X.H.,and Zhao Y.D.,2007,Auxin synthesized by the YUCCA flavin monooxygenases is essential for embryogenesis and leaf formation in Arabidopsis,Plant Cell,19(8):2430-2439
Davies P.J.,ed.,1995,Plant hormones:physiology,biochemistry and molecular biology,Springer Netherlands,Berlin,Germany,pp.1-15
Liu X.Y.,Li J.,Liu M.M.,Yao Q.,and Chen J.Z.,2017,Transcriptome profiling to understand the effect of citrus rootstocks on the growth of'Shatangju'mandarin,PLoS One,12(1):e0169897
Mano Y.,and Nemoto K.,2012,The pathway of auxin biosynthesis in plants,J.Exp.Bot.,63(8):2853-2872
Sambrook J.,and Russell D.W.,eds.,Huang P.T.,trans.,2016,Molecular cloning:a laboratory manuals,Science Press,Beijing,China,pp.865(萨姆布鲁克J.,拉塞尔D.W.,主编,黄培堂,主译,2016,分子克隆实验指南,科学出版社,中国,北京,pp.865)
Santos A.,Ribeiro R.,and CrespíA.L.,2004,Sweet cherry(Prunus avium)growth is mostly affected by rootstock and much less by budding height,New Zeal.J.Crop Hort.,32(3):309-318
Soejima J.,Yoshida Y.,Haniuda T.,Bessho H.,Tsuchiya S.,Masuda T.,Komori S.,Sanada T.,Ito Y.,Sadamori S.,and Kashimura T.,2010,New dwarfing apple rootstocks'JM 1','JM7'and'JM 8',Bull.Natl.Inst.Fruit Tree Sci.,11:1-16
Song C.H.,2017,Study the role of several key IAA meatbolism genes in dwarf rootstock induced scion dwarfing and the role of mi RNA in the spur type apple dwarfing,Dissertation for Ph.D.,Northwest A&F University,Supervisor:Han Y.M.,pp.1-124(宋春晖,2017,苹果IAA代谢几个关键基因在矮化砧木与mi RNA在短枝品种中致矮作用研究,博士学位论文,西北农林科技大学,导师:韩玉明,pp.1-124)
Song C.H.,Zhang D.,Zhang J.,Zheng L.W.,Zhao C.P.,Ma J.J.,An N.,and Han M.Y.,2016,Expression analysis of key auxin synthesis,transport,and metabolism genes in different young dwarfing apple trees,Acta Physiol.Plant.,38(2):1-15
Song J.C,Jiang L.J,and Jameson P.E.,2012,Co-ordinate regulation of cytokinin gene family members during flag leaf and reproductive development in wheat,BMC Plant Biol.,12(1):78-94
Won C.,Shen X.L.,Mashiguchi K.,Zheng Z.Y.,Dai X.H.,Cheng Y.H.,Kasahara H.,Kamiya Y.,Chory J.,and Zhao Y.D.,2011,Conversion of tryptophan to indole-3-acetic acid by TRYPTOPHAN AMINOTRANSFERASES OF ARABIDO-PSIS and YUCCAs in Arabidopsis,Proc.Natl.Acad.Sci.USA,108(45):18518-18523
Zhao Y.,2010,Auxin biosynthesis and its role in plant development,Ann.Rev.Plant Biol.,61(1):49-64
Zhao Y.,Christensen K.S.,Fankhauser C.,Cashman J.R.,Cohen J.D.,Weigel D.,and Chory J.,2001,A role for flavin monooxygenase-like enzymes in auxin biosynthesis,Science,291(5502):306-309