桃GLV多肽受体PpRGI基因家族的生物信息学及表达分析
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摘要
为进一步了解GOLVEN(GLV)多肽在桃果实成熟软化中的分子机制,通过生物信息学鉴定桃基因组中GLV多肽激素受体Pp RGI基因家族,并通过qRT-PCR检测其在溶质型和硬质型桃中的表达水平,检测NAA处理硬质型桃果实和外施多肽处理桃愈伤组织后Pp RGI基因家族的表达水平。结果表明:PpRGI均含有LRR保守结构域,并且在N末端均含有信号肽,聚类分析表明其分为2类。荧光定量PCR分析PpRGI1、PpRGI3、PpRGI4在溶质型桃‘中油桃13号’的不同组织中表达,并且在溶质型桃果实不同发育期中的表达量显著高于硬质型桃果实,其中PpRGI1在不同肉质类型中表达量最高。NAA及多肽处理之后PpRGI1表达均升高。PpRGI1可能参与桃多肽激素的生物信号转导途径。
In order to further understand the molecular mechanism of GLV peptide in peach fruit ripening and softening,the PpRGI gene family was identified in peach genome, and its expression levels during fruit development in melting flesh and stony hard peach were detected by qRT-PCR. Additionally,we also used qRT-PCR to detect the expression level of PpRGI gene family in stony-hard peach fruit under NAA treatment,and in peach callus under exogenous peptide treatment. The results showed that 6 PpRGI candidate genes were found in the peach genome. The PpRGI contained the LRR conserved domain and signal peptide at the N-terminus. The phylogenetic analysis revealed that PpRGI gene family was divided into 2 groups. Expression analysis showed that PpRGI1,PpRGI3 and PpRGI4 were expressed in different tissues of melting-flesh peach and their expression levels in melting-flesh peach fruits were higher than in stony-hard peach fruits. PpRGI1 has the highest expression in different peach types and the expression of PpRGI1 was increased under NAA and peptide treatment. These results suggested that the PpRGI gene family members are highly and structurally conserved and PpRGI1 may be involved in the biosignaling pathway of peach peptide hormones.
In order to further understand the molecular mechanism of GLV peptide in peach fruit ripening and softening,the PpRGI gene family was identified in peach genome, and its expression levels during fruit development in melting flesh and stony hard peach were detected by qRT-PCR. Additionally,we also used qRT-PCR to detect the expression level of PpRGI gene family in stony-hard peach fruit under NAA treatment,and in peach callus under exogenous peptide treatment. The results showed that 6 PpRGI candidate genes were found in the peach genome. The PpRGI contained the LRR conserved domain and signal peptide at the N-terminus. The phylogenetic analysis revealed that PpRGI gene family was divided into 2 groups. Expression analysis showed that PpRGI1,PpRGI3 and PpRGI4 were expressed in different tissues of melting-flesh peach and their expression levels in melting-flesh peach fruits were higher than in stony-hard peach fruits. PpRGI1 has the highest expression in different peach types and the expression of PpRGI1 was increased under NAA and peptide treatment. These results suggested that the PpRGI gene family members are highly and structurally conserved and PpRGI1 may be involved in the biosignaling pathway of peach peptide hormones.
引文
Brandi F,Bar E,Mourgues F,Horváth G,Turcsi E,Giuliano G,Liverani A,Tartarini S,Lewinsohn E,Rosati C.2011.Study of‘Redhaven’peach and its white-fleshed mutant suggests a key role of CCD4 carotenoid dioxygenase in carotenoid and norisoprenoid volatile metabolism.BMC Plant Biology,11(1):24.
B?ttcher C,Keyzers R A,Boss P K,Davies C.2010.Sequestration of auxin by the indole-3-acetic acid-amido synthetase GH3-1 in grape berry(Vitis vinifera L.)and the proposed role of auxin conjugation during ripening.Journal of Experimental Botany,61(13):3615-3625.
Fernandez A,Hilson P,Beeckman T.2013.GOLVEN peptides as important regulatory signalling molecules of plant development.Journal of Experimental Botany,64:5263-5268.
Goodstein D M,Shu S Q,Howson R,Neupane R,Hayes R D,Fazo J,Mitros T,Dirks W,Hellsten U,Putnam N,Rokhsar D S.2012.Phytozome:a comparative platform for green plant genomics.Nucleic Acids Research,40:1178-1186.
Haji T,Yaegaki H,Yamaguchi M,Engei G Z.2001.Changes in ethylene production and flesh firmness of melting,non-melting and stony hard peaches after harvest.Journal of the Japanese Society for Horticultural Science,70(4):458-459.
Haji T,Yaegaki H,Yamaguchi M.2003.Softening of stony hard peach by ethylene and the induction of endogenous ethylene by1-aminocyclopropane-1-carboxylic acid(ACC).Journal of the Japanese Society for Horticultural Science,72(3):212-217.
Livak K J,Schmittgen T D.2001.Analysis of relative gene expression data using Real-time quantitative PCR and the 2-ΔΔCT method.Methods,25(4):402-408.
Matsubayashi Y.2014.Posttranslationally modified small-peptide signals in plants.Annu Rev Plant Biol,65,385-413.
Matsuzaki Y,Ogawa-Ohnishi M,Mori A,Matsubayashi Y.2010.Secreted peptide signals required for maintenance of root stem cell niche in Arabidopsis.Science,329(5995):1065-1067.
Ou Y,Lu X,Zi Q,Xun Q,Zhang J,Wu Y,Shi H,Wei Z,Zhao B,Zhang X,He K,Gou X,Li C,Li J.2016.RGF1INSENSITIVE 1 to 5,a group of LRR receptor-like kinases,are essential for the perception of root meristem growth factor1 in Arabidopsis thaliana.Cell Research,26:686-698.
Pan L,Zeng W,Niu L,Lu Z,Liu H,Cui G.,Zhu Y,Chu J,Li W,Fang W,Cai Z,Li G,Wang Z.2015.PpYUC11,a strong candidate gene for the stony hard phenotype in peach(Prunus persica L.Batsch),participates in IAA biosynthesis during fruit ripening.Journal of Experimental Botany,66(22):7031-7044.
Shinohara H,Mori A,Yasue N,Sumida K,Matsubayashi Y.2016.Identification of three LRR-RKs involved in perception of root meristem growth factor in Arabidopsis.Proc Natl Acad Sci USA,113:3897-3902.
Song W,Liu L,Wang J,Wu Z,Zhang H,Tang J,Lin G,Wang Y,Wen X,Li W,Han Z,Guo H,Chai J.2016.Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth.Cell Research,26:674-685.
Tadiello A,Ziosi V,Negri A S,Noferini M,Fiori G,Busatto N,Espen L,Costa G,Trainotti L.2016.On the role of ethylene,auxin and a golven-like peptide hormone in the regulation of peach ripening.BMC Plant Biology,16:44.
Tatsuki M,Haji T,Yamaguchi M.2006.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,Pp-ACS1,in peach fruit softening.Journal of Experimental Botany,57(6):1281-1289.
Tatsuki M,Nakajima N,Fujii H,Shimada T,Nakano M,Hayashi K I,Hayama H,Yoshioka H,Nakamura Y.2013.Increased levels of IAA are required for system 2 ethylene synthesis causing fruit softening in peach(Prunus persica L.Batsch).Journal of Experimental Botany,64(4):1049-1059.
Verde I,Abbott A G,Scalabrin S,Jung S,Shu S,Marroni F,Zhebentyayeva T,Dettori M T,Grimwood J,Cattonaro F,Zuccolo A,Rossini L,Jenkins J,Vendramin E,Meisel L A,Decroocq V,Sosinski B,Prochnik S,Mitros T,Policriti A,Cipriani G,Dondini L,Ficklin S,Goodstein D M,Xuan P,Del Fabbro C,Aramini V,Copetti D,Gonzalez S,Horner D S,Falchi R,Lucas S,Mica E,Maldonado J,Lazzari B,Bielenberg D,Pirona R,Miculan M,Barakat A,Testolin R,Stella A,Tartarini S,Tonutti P,Arús P,Orellana A,Wells C,Main D,Vizzotto G,Silva H,Salamini F,Schmutz J,Morgante M,Rokhsar D S.2013.The high-quality draft genome of peach(Prunus persica)identifies unique patterns of genetic diversity,domestication and genome evolution.Nature Genetics,45(5):487-494.
Whitford R,Fernandez A,Tejos R,Pérez AC,Kleine-Vehn J,Vanneste S,Drozdzecki A,Leitner J,Abas L,Aerts M,Hoogewijs K,Baster P,Groodt D R,Lin Y C,Storme V,Peer Y V D,Beeckman T,Madder A,Devreese B,Luschnig C,Friml J,Hilson P.2012.GOLVENsecretory peptides regulate auxin carrier turnover during plant gravitropic responses.Dev Cell,22:678-685.
Zeng Wenfang,Pan Lei,Niu Liang,Lu Zhenhua,Cui Guochao,Wang Zhiqiang.2015.Bioinformatics analysis of peach GH3 gene family and expression analysis in fruit development.Acta Horticulturae Sinica,42(5):833-842.曾文芳,潘磊,牛良,鲁振华,崔国朝,王志强.2015.桃GH3基因家族的生物信息学分析及其在果实发育中的表达.园艺学报,42(5):833-842.
Zeng W F,Pan L,Niu L,Lu Z H,Cui G C,Wang Z Q.2015.Characterization of 1-aminocyclopropane-1-carboxylic acid synthase(ACS)genes during nectarine fruit development and ripening.Tree Genetics&Genomes,11(2):1-10.
B?ttcher C,Keyzers R A,Boss P K,Davies C.2010.Sequestration of auxin by the indole-3-acetic acid-amido synthetase GH3-1 in grape berry(Vitis vinifera L.)and the proposed role of auxin conjugation during ripening.Journal of Experimental Botany,61(13):3615-3625.
Fernandez A,Hilson P,Beeckman T.2013.GOLVEN peptides as important regulatory signalling molecules of plant development.Journal of Experimental Botany,64:5263-5268.
Goodstein D M,Shu S Q,Howson R,Neupane R,Hayes R D,Fazo J,Mitros T,Dirks W,Hellsten U,Putnam N,Rokhsar D S.2012.Phytozome:a comparative platform for green plant genomics.Nucleic Acids Research,40:1178-1186.
Haji T,Yaegaki H,Yamaguchi M,Engei G Z.2001.Changes in ethylene production and flesh firmness of melting,non-melting and stony hard peaches after harvest.Journal of the Japanese Society for Horticultural Science,70(4):458-459.
Haji T,Yaegaki H,Yamaguchi M.2003.Softening of stony hard peach by ethylene and the induction of endogenous ethylene by1-aminocyclopropane-1-carboxylic acid(ACC).Journal of the Japanese Society for Horticultural Science,72(3):212-217.
Livak K J,Schmittgen T D.2001.Analysis of relative gene expression data using Real-time quantitative PCR and the 2-ΔΔCT method.Methods,25(4):402-408.
Matsubayashi Y.2014.Posttranslationally modified small-peptide signals in plants.Annu Rev Plant Biol,65,385-413.
Matsuzaki Y,Ogawa-Ohnishi M,Mori A,Matsubayashi Y.2010.Secreted peptide signals required for maintenance of root stem cell niche in Arabidopsis.Science,329(5995):1065-1067.
Ou Y,Lu X,Zi Q,Xun Q,Zhang J,Wu Y,Shi H,Wei Z,Zhao B,Zhang X,He K,Gou X,Li C,Li J.2016.RGF1INSENSITIVE 1 to 5,a group of LRR receptor-like kinases,are essential for the perception of root meristem growth factor1 in Arabidopsis thaliana.Cell Research,26:686-698.
Pan L,Zeng W,Niu L,Lu Z,Liu H,Cui G.,Zhu Y,Chu J,Li W,Fang W,Cai Z,Li G,Wang Z.2015.PpYUC11,a strong candidate gene for the stony hard phenotype in peach(Prunus persica L.Batsch),participates in IAA biosynthesis during fruit ripening.Journal of Experimental Botany,66(22):7031-7044.
Shinohara H,Mori A,Yasue N,Sumida K,Matsubayashi Y.2016.Identification of three LRR-RKs involved in perception of root meristem growth factor in Arabidopsis.Proc Natl Acad Sci USA,113:3897-3902.
Song W,Liu L,Wang J,Wu Z,Zhang H,Tang J,Lin G,Wang Y,Wen X,Li W,Han Z,Guo H,Chai J.2016.Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth.Cell Research,26:674-685.
Tadiello A,Ziosi V,Negri A S,Noferini M,Fiori G,Busatto N,Espen L,Costa G,Trainotti L.2016.On the role of ethylene,auxin and a golven-like peptide hormone in the regulation of peach ripening.BMC Plant Biology,16:44.
Tatsuki M,Haji T,Yamaguchi M.2006.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,Pp-ACS1,in peach fruit softening.Journal of Experimental Botany,57(6):1281-1289.
Tatsuki M,Nakajima N,Fujii H,Shimada T,Nakano M,Hayashi K I,Hayama H,Yoshioka H,Nakamura Y.2013.Increased levels of IAA are required for system 2 ethylene synthesis causing fruit softening in peach(Prunus persica L.Batsch).Journal of Experimental Botany,64(4):1049-1059.
Verde I,Abbott A G,Scalabrin S,Jung S,Shu S,Marroni F,Zhebentyayeva T,Dettori M T,Grimwood J,Cattonaro F,Zuccolo A,Rossini L,Jenkins J,Vendramin E,Meisel L A,Decroocq V,Sosinski B,Prochnik S,Mitros T,Policriti A,Cipriani G,Dondini L,Ficklin S,Goodstein D M,Xuan P,Del Fabbro C,Aramini V,Copetti D,Gonzalez S,Horner D S,Falchi R,Lucas S,Mica E,Maldonado J,Lazzari B,Bielenberg D,Pirona R,Miculan M,Barakat A,Testolin R,Stella A,Tartarini S,Tonutti P,Arús P,Orellana A,Wells C,Main D,Vizzotto G,Silva H,Salamini F,Schmutz J,Morgante M,Rokhsar D S.2013.The high-quality draft genome of peach(Prunus persica)identifies unique patterns of genetic diversity,domestication and genome evolution.Nature Genetics,45(5):487-494.
Whitford R,Fernandez A,Tejos R,Pérez AC,Kleine-Vehn J,Vanneste S,Drozdzecki A,Leitner J,Abas L,Aerts M,Hoogewijs K,Baster P,Groodt D R,Lin Y C,Storme V,Peer Y V D,Beeckman T,Madder A,Devreese B,Luschnig C,Friml J,Hilson P.2012.GOLVENsecretory peptides regulate auxin carrier turnover during plant gravitropic responses.Dev Cell,22:678-685.
Zeng Wenfang,Pan Lei,Niu Liang,Lu Zhenhua,Cui Guochao,Wang Zhiqiang.2015.Bioinformatics analysis of peach GH3 gene family and expression analysis in fruit development.Acta Horticulturae Sinica,42(5):833-842.曾文芳,潘磊,牛良,鲁振华,崔国朝,王志强.2015.桃GH3基因家族的生物信息学分析及其在果实发育中的表达.园艺学报,42(5):833-842.
Zeng W F,Pan L,Niu L,Lu Z H,Cui G C,Wang Z Q.2015.Characterization of 1-aminocyclopropane-1-carboxylic acid synthase(ACS)genes during nectarine fruit development and ripening.Tree Genetics&Genomes,11(2):1-10.
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