Cloning and Expression Pattern of DNA Methyl-binding Domain Gene PsMBD5 in Tree Peony

doi:10.7668/hbnxb.2017.02.011

Abstract

Abstract: The previous results of the transcriptome and expression profiling analysis in tree peony found that a MBD-like fragment was differentially expressed during low temperature induced dormancy release.To investigate the characteristic and putative function of MBD during tree peony bud dormancy release,total 1 204 bp full length cDNA of PsMBD5 was obtained by RACE amplification.The coding fame of 1 056 bp encoded 351 amino acids.The result of bioinformatics analysis showed that the molecular weight of PsMBD5 was 38.127 4 kDa,isoelectric point was 5.14,which indicated that PsMBD5 was an unstable hydrophilic.The result of homology analysis indicated that the similarity between the PsMBD5 and PmMBD5 was the highest with 38.1% identity,and CsMBD5 was the lowest with 25.8%.When compared with 13 kinds of AtMBD,the identity between the PsMBD5 and AtMBD5 was the highest.The expression patterns were analyzed by Real-time quantitative PCR,and the results showed that the transcript level of PsMBD5 at the early stage of flowering in root was the highest,followed by in petals,carpel and sepals,and in stamen was the lowest.During the whole process of chilling fulfillment, PsMBD5 was highly induced by low temperature.The transcript level of PsMBD5 was the highest after 14 d chilling fulfillment,and maintained a high level of transcription after that.These results would provide theoretical basis to validate the regulation mechanism of PsMBD5 during the peony bud dormancy release.

Key words: PsMBD, RACE, Bioinformatics analysis, Expression pattern

CLC Number:

Q78
S685.03

SI Fuhui, ZHANG Yuxi, LIU Chunying, GAI Weiling, ZHAN Xinmei, GAI Shupeng. Cloning and Expression Pattern of DNA Methyl-binding Domain Gene PsMBD5 in Tree Peony[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 66-70. doi: 10.7668/hbnxb.2017.02.011.

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References

[1] Meijón M,Feito I,Valledor L,et al.Dynamics of DNA methylation and Histone H4 acetylation during floral bud differentiation in azalea[J].BMC Plant Biology,2010,10(1):1-14.
[2] 张问,李永春,张艳霞,等.植物甲基结合蛋白(MBD)[J].植物生理学报,2009,45(7):729-732.
[3] Zemach A,Grafi G.Characterization of Arabidopsis thaliana methyl-CpG-binding domain(MBD)proteins[J].The Plant Journal:for Cell and Molecular Biology,2003,34(5):565-572.
[4] Ito M,Koike A,Koizumi N,et al.Methylated DNA-binding proteins from Arabidopsis[J].Plant Physiology,2003,133(4):1747-1754.
[5] Preuss S,Costa-Nunes P,Tucker S,et al.Multimegabase silencing in nucleolar dominance involves siRNA-directed DNA methylation and specific methylcytosine-binding proteins[J].Molecular Cell,2008,32(5):673-684.
[6] Yaish M,Peng M,Rothstein S.AtMBD9 modulates Arabidopsis development through the dual epigenetic pathways of DNA methylation and histone acetylation[J].The Plant Journal:for Cell and Molecular Biology,2009,59(1):123-135.
[7] Berg A,Meza T,Mahić M,et al.Ten members of the Arabidopsis gene family encoding methyl-CpG-binding domain proteins are transcriptionally active and at least one,AtMBD11,is crucial for normal development[J].Nucleic Acids Research,2003,31(18):5291-5304.
[8] Li Y,Deng H,Miao M,et al.Tomato MBD5,a methyl CpG binding domain protein,physically interacting with UV-damaged DNA binding protein-1,functions in multiple processes[J].The New Phytologist,2016,210(1):208-226.
[9] Lang Z,Lei M,Wang X,et al.The methyl-CpG-binding protein MBD7 facilitates active DNA demethylation to limit DNA hyper-methylation and transcriptional gene silencing[J].Molecular Cell,2015,57(6):971-983.
[10] Wang C,Dong X,Jin D,et al.Methyl-CpG-binding domain protein MBD7 is required for active DNA demethylation in Arabidopsis[J].Plant Physiology,2015,167(3):905-914.
[11] Shi R,Zhang J,Li J,et al.Cloning and characterization of TaMBD6 homeologues encoding methyl-CpG-binding domain proteins in wheat[J].Plant Physiology and Biochemistry,2016,109:1-8.
[12] 赵海军.牡丹春节催花技术[M].北京:中国农业出版社,2002.
[13] Gai S P,Zhang Y X,Mu P,et al.Transcriptome analysis of tree peony during chilling requirement fulfillment:Assembling,annotation and markers discovering[J].Gene,2012,497(2):256-262.
[14] Gai S,Zhang Y,Liu C,et al.Transcript profiling of Paoenia ostii during artificial chilling induced dormancy release identifies activation of GA pathway and carbohydrate metabolism[J].PLoS One,2013,8(2):e55297.
[15] 盖树鹏,张风,张玉喜,等.低温解除牡丹休眠进程中基因组DNA甲基化敏感扩增多态性(MSAP)分析[J].农业生物技术学报,2012,20(3):261-267.
[16] 张璐,李玉娥,管世铭,等.牡丹PsAP1基因的表达特性及功能鉴定[J].华北农学报,2015,30(1):84-89.
[17] Livak K J,Schmittgen T D.Analysis of relative gene expression data using Real-time quantitative PCR and the 2(-Delta Delta C(T))Method[J].Methods,2001,25(4):402-408.
[18] 石红梅,战新梅,管世铭,等.牡丹PsWRKY基因的克隆和表达特性分析[J].植物生理学报,2015(10):1743-1748.
[19] Scebba F,Bernacchia G,De Bastiani M,et al. A rabidopsis MBD proteins show different binding specificities and nuclear localization[J].Plant Molecular Biology,2003,53(5):715-731.
[20] Springer N,Kaeppler S.Evolutionary divergence of monocot and dicot methyl-CpG-binding domain proteins[J].Plant Physiology,2005,138(1):92-104.
[21] Questa J,Rius S,Casadevall R,et al.ZmMBD101 is a DNA-binding protein that maintains Mutator elements chromatin in a repressive state in maize[J].Plant,Cell & Environment,2016,39(1):174-184.
[22] Yano A,Kodama Y,Koike A,et al.Interaction between methyl CpG-binding protein and ran GTPase during cell division in tobacco cultured cells[J].Annals of Botany,2006,98(6):1179-1187.

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