唐菖蒲脱落酸受体基因PYL1的克隆及表达分析
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摘要
以唐菖蒲‘超级红’品种为试验材料,采用RT-PCR技术克隆得到其PYL1基因并对其生物信息学进行分析,进一步研究ABA浸泡种球处理后球茎发芽率与PYL1基因表达水平之间的相关性。结果表明:该基因具有687 bp的开放阅读框(ORF),编码228个氨基酸,相对分子质量为25.02 kD,等电点为5.35,具有ABA结合域;其cDNA全长为838 bp,命名为GhPYL1。系统发育树分析表明,GhPYL1与油棕、芭蕉等植物的PYL序列相似性较高。随着ABA处理浓度的逐渐增加,唐菖蒲球茎发芽率逐渐降低,而GhPYL1基因的表达量逐渐升高。球茎发芽率与GhPYL1基因表达水平呈负相关关系,表明该基因可能参与ABA调控的唐菖蒲球茎休眠过程。
Using Gladiolus hybridus 'Su preme Red' variety as the experimental material, the PYL1 gene was cloned by RT-PCR technique and the bioinformatics analysis was carried out. The correlation between corms germination rate and PYL1 gene expression level after ABA soaking corm treatment was further studied. The results showed that the open reading frame(ORF) of this gene was 687 bp, encoding 228 amino acids with the relative molecular mass of 25.02 kD and the isoelectric point of 5.35 with an ABA binding domain. The full length of cDNA was 838 bp, named GhPYL1. Phylogenetic tree analysis showed that GhPYL1 had higher similarity to PYL sequences of plants such as oil palm and plantain. With gradual increase of ABA treatment concentration, the germination rate of gladiolus corms decreased gradually, while the expression of GhPYL1 gene gradually increased. There was a negative correlation between the germination rate of corms and the expression level of GhPYL1 gene, indicating that this gene might be involved in the dormancy process of gladiolus corms regulated by ABA.
Using Gladiolus hybridus 'Su preme Red' variety as the experimental material, the PYL1 gene was cloned by RT-PCR technique and the bioinformatics analysis was carried out. The correlation between corms germination rate and PYL1 gene expression level after ABA soaking corm treatment was further studied. The results showed that the open reading frame(ORF) of this gene was 687 bp, encoding 228 amino acids with the relative molecular mass of 25.02 kD and the isoelectric point of 5.35 with an ABA binding domain. The full length of cDNA was 838 bp, named GhPYL1. Phylogenetic tree analysis showed that GhPYL1 had higher similarity to PYL sequences of plants such as oil palm and plantain. With gradual increase of ABA treatment concentration, the germination rate of gladiolus corms decreased gradually, while the expression of GhPYL1 gene gradually increased. There was a negative correlation between the germination rate of corms and the expression level of GhPYL1 gene, indicating that this gene might be involved in the dormancy process of gladiolus corms regulated by ABA.
引文
Bai G.,Yang D.H.,Zhao Y.,Ha S.,Yang F.,Ma J.,Gao X.S.,Wang Z.M.,and Zhu J.K.,2013,Interactions between soybean ABA receptors and type 2C protein phosphatases,Plant Mol.Biol.,83(6):651-664
Bhujbal G.B.,Chavan N.G.,and Mehetre S.S.,2014,Importance of growth regulator and cold storage treatments for breaking of gladiolus(Gladiolus grandiflorus L.)corm dormancy,Bioscan,9(2):501-505
Cao X.,Costa L.M.,Biderre-Petit C.,Kbhaya B.,Dey N.,Perez P.,Mc Carty D.R.,Gutierrez-Marcos J.F.,and Becraft P.W.,2007,Abscisic acid and stress signals induce viviparous1expression in seed and vegetative tissues of maize,Plant Physiol.,143(2):720-731
Chai Y.M,,Jia H.F.,Li C.L.,Dong Q.H.,and Shen Y.Y.,2011,Fa PYR1 is involved in strawberry fruit ripening,J.Exp.Bot.,62(14):5079-5089
Cutler S.R.,Rodriguez P.L.,Finkelstein R.R.,and Abrams S.R.,2010,Abscisic acid:emergence of a core signaling network,Annu.Rev.Plant Biol.,61:651-679
Gonzalez-Guzman M.,Pizzio G.A.,Antoni R.,Vera-Sirera F.,Merilo E.,Bassel G.W.,Fernández M.A.,Holdsworth M.J.,Perez-Amador M.A.,Kollist H.,and Rodriguez P.L.,2012,Arabidopsis PYR/PYL/RCAR receptors play a major role in quantitative regulation of stomatal aperture and transcriptional response to abscisic acid,Plant Cell,24(6):2483-2496
Gubler F.,Millar A.A.,and Jacobsen J.V.,2005,Dormancy release,ABA and pre-harvest sprouting,Curr.Opin.Plant Biol.,8(2):183-187
Guo J.J.,Yang X.H.,Weston D.J.,and Chen J.G.,2011,Abscisic acid receptors:past,present and future,J.Integr.Plant Biol.,53(6):469-479
Hu S.,Wang F.Z.,Liu Z.N.,Liu Y.P.,and Yu X.L.,2012,ABAsignaling mediated by PYR/PYL/RCAR in plants,Yichuan(Hereditas),34(5):560-572(胡帅,王芳展,刘振宁,刘亚培,余小林,2012,PYR/PYL/RCAR蛋白介导植物ABA的信号转导,遗传,34(5):560-572)
Kim H.,Hwang H.,Hong J.W.,Lee Y.N.,Ahn I.P.,Yoon I.S.,Yoo S.D.,Lee S.,Lee S.C.,and Kim B.G.,2012,A rice orthologue of the ABA receptor,Os PYL/RCAR5,is a positive regulator of the ABA signal transduction pathway in seed germination and early seedling growth,J.Exp.Bot.,63(2):1013-1024
Li D.K.,Zhang L.,Peng M.F.,Hu W.F.,Li Y.,Cai L.,Li X.F.,and Yang Y.,2011,Screening and preliminary study of proteins interacting with ABA receptor RCAR1/PYL9,Sichuan Daxue Xuebao(Journal of Sichuan University(Natural Science Edition)),48(2):486-490(李德款,张亮,彭梅芳,胡伟芳,李鹰,蔡黎,李旭锋,杨毅,2011,ABA受体RCAR1/PYL9相互作用蛋白质的筛选及初步研究,四川大学学报(自然科学版),48(2):486-490)
Li Y.L.,Wang D.D.,Sun C.C.,Hu X.C.,Mu X.Q.,Hu J.J.,Yang Y.Q.,Zhang Y.F.,Xie C.G.,and Zhou X.N.,2017,Molecular characterization of an At PYL1-like protein,Br PYL1,as a putative ABA receptor in Brassica rapa,Biochem.Biophys.Res.Commun.,487(3):684-689
Lin P.C.,Hwang S.G.,Endo A.,Okamoto M.,Koshiba T.,and Cheng W.H.,2007,Ectopic expression of ABSCISIC ACID2/GLUCOSE INSENSITIVE 1 in Arabidopsis promotes seed dormancy and stress tolerance,Plant Physiol.,143(2):745-758
Luo X.,Lu H.,Yuan L.,Jia Y.X.,Wu Y.L.,and Deng Q.X.,2016,Cloning and expression analysis of gibberelin receptor gene in Gladiolus hybridus,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),36(11):2152-2158(罗弦,陆涵,袁雷,贾永霞,武云利,邓群仙,2016,唐菖蒲赤霉素受体基因克隆及表达分析,西北植物学报,36(11):2152-2158)
Luo X.,Yi J.,Zhong X.H.,Lian Q.L.,Khan M.A.,Cao X.,Li X.X.,Gao M.W.,Wu J.,Chen J.,and Yi M.F.,2012,Cloning,characterization and expression analysis of key genes involved in ABA metabolism in Gladiolus cormels during storage,Sci.Hortic.,143(1):115-121
Takeuchi J.,Okamoto M.,Akiyama T.,Mu T.,Yajima S.,Sue M.,Seo M.,Kanno Y.,Kamo T.,Endo A.,Nambara E.,Hirai N.,Ohnishi T.,Cutler S.R.,and Todoroki Y.,2014,Designed abscisic acid analogs as antagonists of PYL-PP2C receptor interactions,Nat.Chem.Biol.,10(6):477-482
Wu J.,Seng S.,Carianopol C.,Sui J.J.,Yang Q.Y.,Zhang F.Q.,Jiang H.R.,He J.N.,and Yi M.F.,2016,Cloning and characterization of a novel Gladiolus hybridus AFP family gene(Gh AFP-like)related to corm dormancy,Biochem.Biophys.Res.Commun.,471(1):198-204
Wu J.,Seng S.S.,Sui J.J.,Vonapartis E.,Luo X.,Gong B.H.,Liu C.,Wu C.Y.,Liu C.,Zhang F.Q.,He J.N.,and Yi M.F.,2015,Gladiolus hybridus ABSCISIC ACID INSENSITIVE5(Gh ABI5)is an important transcription factor in ABA signaling that can enhance Gladiolus corm dormancy and Arabidopsis seed dormancy,Front.Plant Sci.,6:960
Yin P.,Fan H.,Hao Q.,Yuan X.Q.,Wu D.,Pang Y.X.,Yan C.Y.,Li W.Q.,Wang J.W.,and Yan N.,2009,Structural insights into the mechanism of abscisic acid signaling by PYL proteins,Nat.Struct.Mol.Biol.,16(12):1230-1236
Zhang F.Y.,Lu X.,LüZ.Y.,Zhang L.,Zhu M.M.,Jiang W.M.,Wang G.F.,Sun X.F.,and Tang K.X.,2013,Overexpression of the Artemisia orthologue of ABA receptor,Aa PYL9,enhances ABA sensitivity and improves artemisinin content in Artemisia annua L.,PLo S One,8(2):e56697
Bhujbal G.B.,Chavan N.G.,and Mehetre S.S.,2014,Importance of growth regulator and cold storage treatments for breaking of gladiolus(Gladiolus grandiflorus L.)corm dormancy,Bioscan,9(2):501-505
Cao X.,Costa L.M.,Biderre-Petit C.,Kbhaya B.,Dey N.,Perez P.,Mc Carty D.R.,Gutierrez-Marcos J.F.,and Becraft P.W.,2007,Abscisic acid and stress signals induce viviparous1expression in seed and vegetative tissues of maize,Plant Physiol.,143(2):720-731
Chai Y.M,,Jia H.F.,Li C.L.,Dong Q.H.,and Shen Y.Y.,2011,Fa PYR1 is involved in strawberry fruit ripening,J.Exp.Bot.,62(14):5079-5089
Cutler S.R.,Rodriguez P.L.,Finkelstein R.R.,and Abrams S.R.,2010,Abscisic acid:emergence of a core signaling network,Annu.Rev.Plant Biol.,61:651-679
Gonzalez-Guzman M.,Pizzio G.A.,Antoni R.,Vera-Sirera F.,Merilo E.,Bassel G.W.,Fernández M.A.,Holdsworth M.J.,Perez-Amador M.A.,Kollist H.,and Rodriguez P.L.,2012,Arabidopsis PYR/PYL/RCAR receptors play a major role in quantitative regulation of stomatal aperture and transcriptional response to abscisic acid,Plant Cell,24(6):2483-2496
Gubler F.,Millar A.A.,and Jacobsen J.V.,2005,Dormancy release,ABA and pre-harvest sprouting,Curr.Opin.Plant Biol.,8(2):183-187
Guo J.J.,Yang X.H.,Weston D.J.,and Chen J.G.,2011,Abscisic acid receptors:past,present and future,J.Integr.Plant Biol.,53(6):469-479
Hu S.,Wang F.Z.,Liu Z.N.,Liu Y.P.,and Yu X.L.,2012,ABAsignaling mediated by PYR/PYL/RCAR in plants,Yichuan(Hereditas),34(5):560-572(胡帅,王芳展,刘振宁,刘亚培,余小林,2012,PYR/PYL/RCAR蛋白介导植物ABA的信号转导,遗传,34(5):560-572)
Kim H.,Hwang H.,Hong J.W.,Lee Y.N.,Ahn I.P.,Yoon I.S.,Yoo S.D.,Lee S.,Lee S.C.,and Kim B.G.,2012,A rice orthologue of the ABA receptor,Os PYL/RCAR5,is a positive regulator of the ABA signal transduction pathway in seed germination and early seedling growth,J.Exp.Bot.,63(2):1013-1024
Li D.K.,Zhang L.,Peng M.F.,Hu W.F.,Li Y.,Cai L.,Li X.F.,and Yang Y.,2011,Screening and preliminary study of proteins interacting with ABA receptor RCAR1/PYL9,Sichuan Daxue Xuebao(Journal of Sichuan University(Natural Science Edition)),48(2):486-490(李德款,张亮,彭梅芳,胡伟芳,李鹰,蔡黎,李旭锋,杨毅,2011,ABA受体RCAR1/PYL9相互作用蛋白质的筛选及初步研究,四川大学学报(自然科学版),48(2):486-490)
Li Y.L.,Wang D.D.,Sun C.C.,Hu X.C.,Mu X.Q.,Hu J.J.,Yang Y.Q.,Zhang Y.F.,Xie C.G.,and Zhou X.N.,2017,Molecular characterization of an At PYL1-like protein,Br PYL1,as a putative ABA receptor in Brassica rapa,Biochem.Biophys.Res.Commun.,487(3):684-689
Lin P.C.,Hwang S.G.,Endo A.,Okamoto M.,Koshiba T.,and Cheng W.H.,2007,Ectopic expression of ABSCISIC ACID2/GLUCOSE INSENSITIVE 1 in Arabidopsis promotes seed dormancy and stress tolerance,Plant Physiol.,143(2):745-758
Luo X.,Lu H.,Yuan L.,Jia Y.X.,Wu Y.L.,and Deng Q.X.,2016,Cloning and expression analysis of gibberelin receptor gene in Gladiolus hybridus,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),36(11):2152-2158(罗弦,陆涵,袁雷,贾永霞,武云利,邓群仙,2016,唐菖蒲赤霉素受体基因克隆及表达分析,西北植物学报,36(11):2152-2158)
Luo X.,Yi J.,Zhong X.H.,Lian Q.L.,Khan M.A.,Cao X.,Li X.X.,Gao M.W.,Wu J.,Chen J.,and Yi M.F.,2012,Cloning,characterization and expression analysis of key genes involved in ABA metabolism in Gladiolus cormels during storage,Sci.Hortic.,143(1):115-121
Takeuchi J.,Okamoto M.,Akiyama T.,Mu T.,Yajima S.,Sue M.,Seo M.,Kanno Y.,Kamo T.,Endo A.,Nambara E.,Hirai N.,Ohnishi T.,Cutler S.R.,and Todoroki Y.,2014,Designed abscisic acid analogs as antagonists of PYL-PP2C receptor interactions,Nat.Chem.Biol.,10(6):477-482
Wu J.,Seng S.,Carianopol C.,Sui J.J.,Yang Q.Y.,Zhang F.Q.,Jiang H.R.,He J.N.,and Yi M.F.,2016,Cloning and characterization of a novel Gladiolus hybridus AFP family gene(Gh AFP-like)related to corm dormancy,Biochem.Biophys.Res.Commun.,471(1):198-204
Wu J.,Seng S.S.,Sui J.J.,Vonapartis E.,Luo X.,Gong B.H.,Liu C.,Wu C.Y.,Liu C.,Zhang F.Q.,He J.N.,and Yi M.F.,2015,Gladiolus hybridus ABSCISIC ACID INSENSITIVE5(Gh ABI5)is an important transcription factor in ABA signaling that can enhance Gladiolus corm dormancy and Arabidopsis seed dormancy,Front.Plant Sci.,6:960
Yin P.,Fan H.,Hao Q.,Yuan X.Q.,Wu D.,Pang Y.X.,Yan C.Y.,Li W.Q.,Wang J.W.,and Yan N.,2009,Structural insights into the mechanism of abscisic acid signaling by PYL proteins,Nat.Struct.Mol.Biol.,16(12):1230-1236
Zhang F.Y.,Lu X.,LüZ.Y.,Zhang L.,Zhu M.M.,Jiang W.M.,Wang G.F.,Sun X.F.,and Tang K.X.,2013,Overexpression of the Artemisia orthologue of ABA receptor,Aa PYL9,enhances ABA sensitivity and improves artemisinin content in Artemisia annua L.,PLo S One,8(2):e56697