菠萝叶刺形成相关基因AcSPL14克隆与载体构建
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摘要
本研究以菠萝叶片为材料,从中克隆到一个可能与菠萝叶刺形成相关的基因AcSPL14 (Ananas comosus squamosa promoter-binding-like protein 14)。该基因cDNA全长为1 330 bp,开放阅读框为1 086 bp,编码362个氨基酸。结构域分析结果显示AcSPL14蛋白序列具有SPL蛋白家族共有的功能结构域——SBP(SQUAMOSA promoter binding protein-like)结构域,且与石刁柏(Asparagus officinalis)、铁皮石斛(Dendrobium catenatum)、月季(Rosa chinensis)和麻风树(Jatropha curcas)的SBP序列相似性分别为53%、51%、47%和46%。在此基础上,本研究进一步构建了p BI121-AcSPL14过表达载体,获得了工程菌。本研究为AcSPL14在菠萝中的生物学功能鉴定提供了理论依据,对深入研究菠萝叶刺发生的分子机理具有重要意义。
In this study, gene AcSPL14(Ananas comosus squamosa promoter-binding-like protein 14) was cloned from the leaves of Ananas comosus, which was considerably related to the phenotype of leaf spine in pineapple.The cDNA full length of AcSPL14 was 1 330 bp, with the open reading frame(ORF) of 1 086 bp, and encoded a protein consisted of 362 amino acid. The domain analysis showed that the AcSPL14 protein sequence possessed a common functional domain of the SPL protein family, containing a typical SBP(SQUAMOSA promoter binding protein-like) domain. Phylogenetic analysis indicated that protein sequences of AcSPL14 was 53%, 51%, 47% and46% homologous with those of Asparagus officinalis, Dendrobium catenatum, Rosa chinensis and Jatropha curcas,respectively. Furthermore, a pBI121-AcSPL14 overexpression vector was constructed in this study and the engineering bacteria was obtained. This study could provide a theoretical basis for the identification of the biological function of AcSPL14 in pineapple, and was of great significance for the in-depth study of the molecular mechanism of pineapple leaf thorn.
In this study, gene AcSPL14(Ananas comosus squamosa promoter-binding-like protein 14) was cloned from the leaves of Ananas comosus, which was considerably related to the phenotype of leaf spine in pineapple.The cDNA full length of AcSPL14 was 1 330 bp, with the open reading frame(ORF) of 1 086 bp, and encoded a protein consisted of 362 amino acid. The domain analysis showed that the AcSPL14 protein sequence possessed a common functional domain of the SPL protein family, containing a typical SBP(SQUAMOSA promoter binding protein-like) domain. Phylogenetic analysis indicated that protein sequences of AcSPL14 was 53%, 51%, 47% and46% homologous with those of Asparagus officinalis, Dendrobium catenatum, Rosa chinensis and Jatropha curcas,respectively. Furthermore, a pBI121-AcSPL14 overexpression vector was constructed in this study and the engineering bacteria was obtained. This study could provide a theoretical basis for the identification of the biological function of AcSPL14 in pineapple, and was of great significance for the in-depth study of the molecular mechanism of pineapple leaf thorn.
引文
Bowman J.L.,2000,Axia l patterning in leaves and other lateral organs,Curr.Opin.Genet.Dev.,10(4):399-404
Bowman J.L.,Eshed Y.,and Baum S.F.,2002,Establishment of polarity in angiosperm lateral organs,Trends Genet.,18(3):134-141
Cardon G.H.,Hhmann S.,Nettesheim K.,Saedler H.,and Huijser P.,1997,Functional analysis of the Arabidopsis thaliana SBP-box gene SPL3:a novel gene involved in the floral transition,Plant J.,12(2):367-377
Eriksson E.M.,Bovy A.,Manning K.,Harrison L.,Andrews J.,De S.J.,Tucker G.A.,and Seymour G.B.,2004,Effect of the Colorless non-ripening mutation on cell wall biochemistry and gene expression during tomato fruit development and ripening,Plant Physiol.,136(4):4184-4197
Gou J.Y.,Felippes F.F.,Liu C.J.,Weigel D.,and Wang J.W.,2011,Negative regulation of anthocyanin biosynthesis in Arabidopsis by a miR156-targeted SPL transcription factor,Plant Cell,23(4):1512-1522
Hasson A.,Blein T.,and Laufs P.,2010,Leaving the meristem behind:the genetic and molecular control of leaf patterning and morphogenesis,C.R.Biol.,333(4):350-360
Hou H.M.,Li J.,Gao M.,Singer S.D.,Wang H.,Mao L.Y.,Fei Z.J.,and Wang X.P.,2013,Genomic organization,phylogenetic comparison and differential expression of the SBP-Box family genes in grape,PLo S One,8(3):e59358
Hudson A.,1999,Axioms and axes in leaf formation?Curr.Opin.Plant Biol.,2(1):56-60
Huijser P.,Klein J.,Lnnig W.E.,Meijer H.,Saedler H.,and Sommer H.,1992,Bracteomania,an inflorescence anomaly,is caused by the loss of function of the MADS-box gene squamosa in Antirrhinum majus,EMBO J.,11(4):1239-1249
Hultquist J.F.,and Dorweiler J.E.,2008,Feminized tassels of maize mop1 and ts1 mutants exhibit altered levels of miR156 and specific SBP-box genes,Planta,229(1):99-113
Jiao Y.,Wang Y.,Xue D.,Wang J.,Yan M.,Liu G.,Dong G.,Zeng D.,Lu Z.,Qian Q.,and Li J.,2010,Regulation of OsS-PL14 by Osmi R1 56 defines ideal plant architecture in rice,Nat.Genet.,42(6):541-544
Jung J.H.,Seo P.J.,Kang S.K.,and Park C.M.,2011,mi R172 signals are incorporated into the miR156 signaling pathway at the SPL3/4/5 genes in Arabidopsis developmental transitions,Plant Mol.Biol.,76(1-2):35-45
Li J.,Hou H.,Li X.,Jiang X.,Yin X.,Gao H.,Zheng Y.,Bassett C.L.,and Wang X.,2013,Genome-wide identification and analysis of the SBP-box family genes in apple(Malus×domestica Borkh.),Plant Physiol.Biochem.,70(1):100-114
Salinas M.,Xing S.,Hhmann S.,Berndtgen R.,and Huijser P.,2012,Genomic organization,phylogenetic comparison and differential expression of the SBP-box family of transcription factors in tomato,Planta,235(6):1171-1184
Shikata M.,Koyama T.,Mitsuda N.,and Ohme-Takagi M.,2009,Arabidopsis SBP-Box genes SPL10,SPL11 and SPL2 control morphological change in association with shoot maturation in the reproductive phase,Plant Cell Physiol.,50(12):2133-2145
Stone J.M.,Liang X.,Nekl E.R.,and Stiers J.J.,2005,Arabidopsis AtSPL14,a plant-specific SBP-domain transcription factor,participates in plant development and sensitivity to fumonisin B1,Plant J.,41(5):744-754
Wang J.W.,and Weigel D.,2008,Dual effects of miR156-targeted SPL genes and CYP78A5/KLUH on plastochron length and organ size in Arabidopsis thaliana,Plant Cell,20(5):1231-1243
Wang L.,and Zhang Q.,2017,Boosting rice yield by fine-tuning SPL gene expression,Trends Plant Sci.,22(8):643-646
Wu G.,and Poethig R.S.,2006,Temporal regulation of shoot development in Arabidopsis thaliana by miR156 and its target SPL3,Development,133(18):3539-3547
Zhang H.X.,Jin J.H.,He Y.M.,Lu B.Y.,Li D.W.,Chai W.G.,Khan A.,and Gong Z.H.,2016,Genome-wide identification and analysis of the SBP-Box family genes under Phytophthora capsici stress in pepper(Capsicum annuum L.),Front.Plant Sci.,7(1):504
Zhang X.H.,Dou L.L.,Pang C.Y.,Song M.Z.,Wei H.L.,Fan S.L.,Wang C.S.,and Yu S.X.,2015,Genomic organization,differential expression,and functional analysis of the SPL gene family in Gossypium hirsutum,Mol.Genet.Genomics,290(1):115-126
Zhang Y.,Schwarz S.,Saedler H.,and Huijser P.,2007,SPL8,a local regulator in a subset of gibberellin-mediated developmental processes in Arabidopsis,Plant Mol.Biol.,63(3):429-439
Bowman J.L.,Eshed Y.,and Baum S.F.,2002,Establishment of polarity in angiosperm lateral organs,Trends Genet.,18(3):134-141
Cardon G.H.,Hhmann S.,Nettesheim K.,Saedler H.,and Huijser P.,1997,Functional analysis of the Arabidopsis thaliana SBP-box gene SPL3:a novel gene involved in the floral transition,Plant J.,12(2):367-377
Eriksson E.M.,Bovy A.,Manning K.,Harrison L.,Andrews J.,De S.J.,Tucker G.A.,and Seymour G.B.,2004,Effect of the Colorless non-ripening mutation on cell wall biochemistry and gene expression during tomato fruit development and ripening,Plant Physiol.,136(4):4184-4197
Gou J.Y.,Felippes F.F.,Liu C.J.,Weigel D.,and Wang J.W.,2011,Negative regulation of anthocyanin biosynthesis in Arabidopsis by a miR156-targeted SPL transcription factor,Plant Cell,23(4):1512-1522
Hasson A.,Blein T.,and Laufs P.,2010,Leaving the meristem behind:the genetic and molecular control of leaf patterning and morphogenesis,C.R.Biol.,333(4):350-360
Hou H.M.,Li J.,Gao M.,Singer S.D.,Wang H.,Mao L.Y.,Fei Z.J.,and Wang X.P.,2013,Genomic organization,phylogenetic comparison and differential expression of the SBP-Box family genes in grape,PLo S One,8(3):e59358
Hudson A.,1999,Axioms and axes in leaf formation?Curr.Opin.Plant Biol.,2(1):56-60
Huijser P.,Klein J.,Lnnig W.E.,Meijer H.,Saedler H.,and Sommer H.,1992,Bracteomania,an inflorescence anomaly,is caused by the loss of function of the MADS-box gene squamosa in Antirrhinum majus,EMBO J.,11(4):1239-1249
Hultquist J.F.,and Dorweiler J.E.,2008,Feminized tassels of maize mop1 and ts1 mutants exhibit altered levels of miR156 and specific SBP-box genes,Planta,229(1):99-113
Jiao Y.,Wang Y.,Xue D.,Wang J.,Yan M.,Liu G.,Dong G.,Zeng D.,Lu Z.,Qian Q.,and Li J.,2010,Regulation of OsS-PL14 by Osmi R1 56 defines ideal plant architecture in rice,Nat.Genet.,42(6):541-544
Jung J.H.,Seo P.J.,Kang S.K.,and Park C.M.,2011,mi R172 signals are incorporated into the miR156 signaling pathway at the SPL3/4/5 genes in Arabidopsis developmental transitions,Plant Mol.Biol.,76(1-2):35-45
Li J.,Hou H.,Li X.,Jiang X.,Yin X.,Gao H.,Zheng Y.,Bassett C.L.,and Wang X.,2013,Genome-wide identification and analysis of the SBP-box family genes in apple(Malus×domestica Borkh.),Plant Physiol.Biochem.,70(1):100-114
Salinas M.,Xing S.,Hhmann S.,Berndtgen R.,and Huijser P.,2012,Genomic organization,phylogenetic comparison and differential expression of the SBP-box family of transcription factors in tomato,Planta,235(6):1171-1184
Shikata M.,Koyama T.,Mitsuda N.,and Ohme-Takagi M.,2009,Arabidopsis SBP-Box genes SPL10,SPL11 and SPL2 control morphological change in association with shoot maturation in the reproductive phase,Plant Cell Physiol.,50(12):2133-2145
Stone J.M.,Liang X.,Nekl E.R.,and Stiers J.J.,2005,Arabidopsis AtSPL14,a plant-specific SBP-domain transcription factor,participates in plant development and sensitivity to fumonisin B1,Plant J.,41(5):744-754
Wang J.W.,and Weigel D.,2008,Dual effects of miR156-targeted SPL genes and CYP78A5/KLUH on plastochron length and organ size in Arabidopsis thaliana,Plant Cell,20(5):1231-1243
Wang L.,and Zhang Q.,2017,Boosting rice yield by fine-tuning SPL gene expression,Trends Plant Sci.,22(8):643-646
Wu G.,and Poethig R.S.,2006,Temporal regulation of shoot development in Arabidopsis thaliana by miR156 and its target SPL3,Development,133(18):3539-3547
Zhang H.X.,Jin J.H.,He Y.M.,Lu B.Y.,Li D.W.,Chai W.G.,Khan A.,and Gong Z.H.,2016,Genome-wide identification and analysis of the SBP-Box family genes under Phytophthora capsici stress in pepper(Capsicum annuum L.),Front.Plant Sci.,7(1):504
Zhang X.H.,Dou L.L.,Pang C.Y.,Song M.Z.,Wei H.L.,Fan S.L.,Wang C.S.,and Yu S.X.,2015,Genomic organization,differential expression,and functional analysis of the SPL gene family in Gossypium hirsutum,Mol.Genet.Genomics,290(1):115-126
Zhang Y.,Schwarz S.,Saedler H.,and Huijser P.,2007,SPL8,a local regulator in a subset of gibberellin-mediated developmental processes in Arabidopsis,Plant Mol.Biol.,63(3):429-439