牡丹DNA甲基结合域蛋白基因<em>PsMBD5</em>的克隆和表达特性分析

doi:10.7668/hbnxb.2017.02.011

华北农学报 ›› 2017, Vol. 32 ›› Issue (2): 66-70. doi: 10.7668/hbnxb.2017.02.011

所属专题： 生物技术；

牡丹DNA甲基结合域蛋白基因PsMBD5的克隆和表达特性分析

司福会¹, 张玉喜¹, 刘春英¹, 盖伟玲², 战新梅¹, 盖树鹏¹

1. 青岛农业大学生命科学学院, 山东省高校植物生物技术重点实验室, 山东青岛 266109;
2. 青岛农业大学农学与植保学院, 山东青岛 266109

收稿日期:2017-01-06 出版日期:2017-04-28
通讯作者: 盖树鹏(1974-),男,山东莱阳人,教授,博士,主要从事牡丹分子生物学研究。
作者简介:司福会(1990-),女,山东菏泽人,在读硕士,主要从事牡丹分子生物学研究。
基金资助:
国家自然基金面上项目（31471908；31372104）

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

SI Fuhui¹, ZHANG Yuxi¹, LIU Chunying¹, GAI Weiling², ZHAN Xinmei¹, GAI Shupeng¹

1. College of Life Sciences, Qingdao Agricultural University, Key Lab of Plant Biotechnology in Universities of Shandong Province, Qingdao 266109, China;
2. College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao 266109, China

Received:2017-01-06 Published:2017-04-28

摘要/Abstract

摘要： 前期通过转录组结合表达谱分析发现一类MBD片段在低温解除牡丹花芽内休眠中的差异表达，为了进一步研究MBD基因特征及其在牡丹休眠解除中的作用，利用RACE扩增获得1 204 bp PsMBD5全长cDNA，其中编码框1 056 bp，推测编码351个氨基酸。生物信息学分析结果显示，PsMBD5蛋白分子量约为38.127 4 kDa，等电点为5.14，不稳定系数为44.27，推测该蛋白为不稳定蛋白。同源性分析表明，牡丹PsMBD5与梅PmMBD5的同源性最高，为38.1%，与亚麻荠CsMBD5的同源性最低，为25.8%，与13种拟南芥MBD蛋白同源性最高的是AtMBD5。实时荧光定量PCR分析表明，PsMBD5基因在牡丹不同组织中均有表达，在初花期牡丹的根中转录水平最高，在花瓣、心皮和萼片中次之，在雄蕊中最低；PsMBD5基因在牡丹花芽中受低温诱导，且转录水平在低温处理14 d时达到最大值，之后维持较高的转录水平。研究结果为进一步解析PsMBD5基因对牡丹花芽的休眠解除的调控机制奠定基础。

关键词: PsMBD, RACE, 生物信息学分析, 表达特性

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

中图分类号:

Q78
S685.03

司福会, 张玉喜, 刘春英, 盖伟玲, 战新梅, 盖树鹏. 牡丹DNA甲基结合域蛋白基因PsMBD5的克隆和表达特性分析[J]. 华北农学报, 2017, 32(2): 66-70. doi: 10.7668/hbnxb.2017.02.011.

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.

http://www.hbnxb.net/CN/Y2017/V32/I2/66

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牡丹DNA甲基结合域蛋白基因PsMBD5的克隆和表达特性分析

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牡丹DNA甲基结合域蛋白基因PsMBD5的克隆和表达特性分析

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

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