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作物学报 ›› 2014, Vol. 40 ›› Issue (03): 381-389.doi: 10.3724/SP.J.1006.2014.00381

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

花生质体型酰基载体蛋白基因5'侧翼调控序列的克隆与分析

单雷1,2,唐桂英1,徐平丽1,赵学彬1,3,柳展基1   

  1. 1 山东省农业科学院高新技术研究中心 / 山东省作物遗传改良与生态生理重点实验室, 山东济南 250100; 2 山东大学农学院, 山东济南 250100; 3 山东师范大学生命科学学院, 山东济南 250014
  • 收稿日期:2013-07-03 修回日期:2013-10-31 出版日期:2014-03-12 网络出版日期:2014-01-16
  • 通讯作者: 单雷, E-mail: shlei1025@sina.com, Tel: 0531-83179435
  • 基金资助:

    本研究由山东省自然科学基金项目(ZR2009DM013)和山东省科技发展计划项目(2012GNC1101)资助。

Cloning and Analysis of 5′ Flanking Regions of Arachisis hypogaea L. Genes Encoding Plastidial Acyl Carrier Protein

SHAN Lei1,2,*,TANG Gui-Ying1,XU Ping-Li1,ZHAO Xue-Bin1,3,LIU Zhan-Ji1   

  1. 1 Hi-Tech Research Centre, Shandong Academy of Agricultural Sciences/Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan 250100, China; 2 College of Agriculture, Shandong University, Jinan 250100, China; 3 College of Life Science, Shandong Normal University, Jinan 250014, China
  • Received:2013-07-03 Revised:2013-10-31 Published:2014-03-12 Published online:2014-01-16
  • Contact: 单雷, E-mail: shlei1025@sina.com, Tel: 0531-83179435

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摘要：

采用染色体步移技术分别克隆3个花生质体型酰基载体蛋白(ACP)基因的5¢侧翼调控区序列, AhACP1、AhACP4和AhACP5基因5¢上游序列分别为535、1400和1180 bp; 利用5¢RACE方法确定了这3个基因的转录起始位点, 分别位于起始密码ATG上游–71 bp、–92 bp和–71 bp处。利用生物信息学软件分析了花生ACPs启动子区包含的主要调控元件, 发现尽管花生AhACP4和AhACP5基因在根、茎、叶、花和不同发育期种子中的基本表达模式相似, 但它们的启动子中包含各自特有的顺式元件, AhACP4启动子区包含根或芽顶端分生组织表达调控元件WUS, 而AhACP5启动子区则含有侧芽萌动和伸展所需的多个关键调控元件E2FB、TELO BOX和UP1, 推测它们的表达具有组织和发育阶段特异性。在进化上, 花生AhACP4与拟南芥AtACP4可能为直系同源基因, 但它们的表达模式产生了分歧, AhACP4为组成型表达, AtACP4主要在叶中表达; 与AtACP4启动子相比, 花生AhACP4启动子区中参与光调控相关元件明显减少。

关键词: 质体型酰基载体蛋白, 启动子, 调控元件, 花生

Abstract:

We cloned the 5′ flanking fragments of three peanut plastidial acyl carrier protein (ACP) genes AhACP1, AhACP4,and AhACP5 by chromosome walking method, and the fragments sizes were 535, 1400, and 1180 bp, respectively. Using 5′ RACE (5′ Rapid Amplification of cDNA End), the transcription start sites of the three genes were localized on –71, –92, and –71 bp from the translation initiation codon ATG, respectively. The crucial regulation elements in promoters of three peanut ACP genes and four Arabidopsis ACP genes were further analyzed with softwares PLACE and Plant CARE (http://www.dna.affrc.go.jp/PLACE; http://bioinformatics.psb.ugebp.be/webtools/plabpcare/html), showing that although AhACP4 and AhACP5 had similar expression patterns in root, stem, leaf, flower and seed at different development stages, their promoter regions contained their specific cis-elements. For instance, the promoter region of AhACP4 consisted of the regulation element WUS expressingin meristems of root or shoot primordia while that of AhACP5 had several key regulation elements such as E2FB, TELO BOX, and UP1 required in sprouting and expansion of axillary shoots, suggesting that their expression patterns may be various in different tissues and development stages. This study also indicated that the expression profiles of orthologous genes peanut AhACP4 and Arabidopsis AtACP4 diverged in evolution, and the AhACP4 gene expressed in a constitutive pattern while the AtACP4 expressed mainly in leaf, and in comparison with the promoter region of AtACP4, that of AhACP4 contained fewer light regulation elements.

Key words:  Plastidial acyl carrier protein (ACP), Promoter, Regulation element, Peanut (Arachis hypogaea L.)

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