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作物学报 ›› 2010, Vol. 36 ›› Issue (06): 945-952.doi: 10.3724/SP.J.1006.2010.00945

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米干旱诱导表达基因ZmCKS2的克隆与表达分析

张中保1,卢敏1,李会勇1,2,张登峰1,刘颖慧1,3,石云素1,宋燕春1,王天宇1,*,黎裕1,*   

  1. 1中国农业科学院作物科学研究所，北京100081；2河南省农业科学院粮食作物研究所，河南郑州450002；3河北北方学院，河北张家口075000
  • 收稿日期:2010-01-29 修回日期:2010-03-04 出版日期:2010-06-12 网络出版日期:2010-04-14
  • 通讯作者: 王天宇, E-mail: wangtianyu@263.net; 黎裕, E-mail: yuli@mail.caas.net.cn
  • 基金资助:

    本研究由国家重点基础研究发展计划（973计划）项目（2006CB101700），国家高技术研究发展计划（863计划）项目（2006AA10Z188）和国家自然科学基金重点项目（30730063）资助。

Isolation and Expression Analysis of a Drought-Induced Gene “ZmCKS2” in Maize (Zea mays L.)

ZHANG Zhong-Bao1,LEI Min1,LI Hui-Yong1,2,ZHANG De-Feng1,LIU Ying-Hui1,3,SHI Yun-Su1, SONG Yan-Chun1,WANG Tian-Yu1,*,LI Yu1,*   

  1. 1Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2Cereal Crops Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;3Hebei North University,Zhangjiakou 075000,China
  • Received:2010-01-29 Revised:2010-03-04 Published:2010-06-12 Published online:2010-04-14
  • Contact: WANG Tian-Yu, E-mail: wangtianyu@263.net;LI Yu, E-mail: yuli@mail.caas.net.cn

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2594

被引次数

5

摘要：

在本实验室前期研究的干旱胁迫相关的抑制差减文库(SSH)中，发现一个玉米干旱胁迫诱导表达的EST序列，与拟南芥AtCKS2和AtCKS1基因有较高的同源性。应用生物信息学手段和同源克隆的方法，分离得到该基因，命名为ZmCKS2。序列分析表明该基因开放阅读框区267 bp，编码88个氨基酸。ZmCKS2蛋白含有真核生物中高度保守的CKS (cyclin-dependent kinase regulatory subunit)结构域。应用在线分析软件预测该基因上游2 kb序列表明，该序列具有启动子的核心序列和增强子序列，同时还具有与干旱等多种逆境胁迫相关的调控序列。应用实时荧光定量PCR分析表明该基因在幼穗中表达量最高，且受干旱和MeJA诱导上调表达，受冷和外源ABA诱导下调表达。

关键词: ZmCKS2, 逆境胁迫, 表达模式, 启动子, 玉米

Abstract:

Plants respond to abiotic and biotic stresses through a number of biochemical and physiological reactions. Stresses inducible genes can be classified into two groups, i.e. functional genes that directly protect against stresses and regulator genes that regulate gene expression and signal transduction in the stress response. In our previous study, an EST from our drought related SSH library (suppression subtractive hybridization library) was obtained, which had a high similarity to AtCKS2in Arabidopsis. In the present study we cloned the cDNA homologous to CKS2, named ZmCKS2, by using in silico and homology-based cloning techniques. ZmCKS2 had an ORF of 267 bp and encoded 88 amino acids. The deduced protein of ZmCKS2 was predicted to contain a CKS (cyclin-dependent kinase regulatory subunit) structural domain, which was highly conserved in eukaryotes kingdom. The promoter of ZmCKS2 (about 2 kb upstream of ZmCKS2)was predicted to contain important regulatory elements including core promoter elements and drought, high salt, cold, ABA, light regulation, copper and oxygen responsive elements. Real-time quantitative PCR (qRT-PCR) analysis revealed that ZmCKS2 had different expression patterns in different organs. It was highly expressed in ears while its expression was very low in roots and leaves at seedling stage. qRT-PCR was also performed to investigate the expression profiles of the ZmCKS2 under different abiotic stresses such as drought, low temperature, high salt, MeJA (methyl jasmonate), and exogenous ABA (abscisic acid). Under drought or MeJA treatment, ZmCKS2 showed an up-regulated expression pattern. However, under cold or ABA treatment, the gene showed a down-regulation expression pattern. Under NaCl stress ZmCKS2 showed no obvious change. These results revealed that ZmCKS2 might be involved in multiple pathways of plants responding to different environmental conditions.

Key words: ZmCKS2, Abiotic stresses, Expression profiles, Promoter, Zea mays

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