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作物学报 ›› 2009, Vol. 35 ›› Issue (8): 1410-1417.doi: 10.3724/SP.J.1006.2009.01410

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米蔗糖转运蛋白基因ZmERD6 cDNAs 的克隆与逆境条件下的表达

马小龙1,2,刘颖慧2,3,**,袁祖丽1,*,石云素2,宋燕春2,王天宇2,黎裕2,*   

  1. 1河南农业大学生命科学学院,河南郑州450032;2中国农业科学院作物科学研究所,北京100081;3河北北方学院,河北张家口075000
  • 收稿日期:2009-02-16 修回日期:2009-04-22 出版日期:2009-08-12 网络出版日期:2009-06-10
  • 通讯作者: 黎裕,E-mail: yuli@mail.caas.net.cn;袁祖丽,E-mail: zuliyuan@yahoo.com.cn
  • 基金资助:

    本研究由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项目(30571133)资助。

Cloning of cDNAs for a Noval Sugar Transporter Gene,ZmERD6,from Maize and Its Expression Analysis under Abiotic Stresses

MA Xiao-Long1,2, LIU Ying-Hui2,3,**, YUAN Zu-Li1,*, SHI Yun-Su2, SONG Yan-Chun2, WANG Tian-Yu2, and LI Yu2,*   

  1. 1 College of Life Science, Henan Agricultural University, Zhengzhou 450002, China; 2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 Hebei North University, Zhangjiakou 075000, China
  • Received:2009-02-16 Revised:2009-04-22 Published:2009-08-12 Published online:2009-06-10
  • Contact: LI Yu,E-mail: yuli@mail.caas.net.cn;YUAN Zu-Li,E-mail: zuliyuan@yahoo.com.cn

摘要:

在前期的研究中发现一个玉米苗期早期应答干旱的EST序列与拟南芥ERD6序列同源性较高。本研究应用电子克隆与同源克隆技术分离出这个基因,并命名为ZmERD6。研究表明ZmERD6具有大小两个转录本,分别被命名为ZmERD6-LZmERD6-SZmERD6-L开放阅读框1 515 bp,编码505个氨基酸,ZmERD6-S开放阅读框1 386 bp,编码463个氨基酸。序列分析表明,ZmERD6-LZmERD6-S蛋白结构中含有两个MFS (major facilitator super-family)结构域,属于MFS家族的蔗糖转运子(sugar transporter)亚族。跨膜结构预测表明两个蛋白都具有多个跨膜区,ZmERD6-L蛋白含12个而ZmERD6-S11个跨膜区。利用半定量RT-PCR分析ZmERD6ABA、干旱、盐和冷胁迫处理以及不同组织中的表达情况,表明ZmERD6基因在不同胁迫下能被诱导表达,在不同组织中表达有差异。通过在玉米基因组数据库中的查询和比对获得ZmERD6基因上游2.5 kb的序列,生物信息学预测表明该序列具有启动子的核心序列及上游增强子序列、抑制子序列,同时还具有冷、茉莉酸甲酯等激素调控序列。

关键词: ZmERD6, 蔗糖转运蛋白, 启动子, 基因表达, 玉米

Abstract:

Drought is one of the most important limiting factors for crop yield in the majority of agricultural regions around the world. Plants respond to drought stress at physiological, cellular and molecular levels. Carbohydrate substances as a nutrient and signal substances play an important role in plants throughout the life course. Carbohydrate has a variety of functions, such as providing energy for the cell life, a framework for proteins and nucleic acid molecules, and raw materials for new cells, regulating osmotically in plants to enhance tolerance, activating different signal transduction pathways and inhibition or activation of certain plant genes which regulate many physiological processes. Previous studies reported that ERD6 is a sugar transporter which widely exists in plants, belonging to Major Facilitator Super-family (MFS) which has a typical structure of MFS domain and sugar transport proteins signature. An EST with high similarity to ERD6 in Arabidopsis was found previously in maize. In the present study cDNAs for the gene homologous to ERD6, designated ZmERD6, was obtained through in silico and homology-based cloning techniques. ZmERD6 had two transcripts, i.e. ZmERD6-L (the large one) and ZmERD6-S (the small one). ZmERD6-L had an ORF of 1 515 bp and encoded 505 amino acids (AA) while ZmERD6-S had an ORF of 1 386 bp and encoded 463 AA. The deduced protein of ZmERD6-L and ZmERD6-S was predicted to contain 12 and 11 membrane spanning helices, respectively. Both of the two proteins had two MFS structural domain belonging to the sugar transporter sub-family of the MFS. Reverse transcription-PCR analysis was performed to investigate the expression pattern of the ZmERD6 in maize under various abiotic stresses. ZmERD6 was expressed at different stages through maize development and was also induced by different abiotic stresses. The promoter of ZmERD6 was cloned, which was about 2.5 kb upstream of ZmERD6 and was predicted to contain important regulatory elements including core promoter elements, enhancer elements, repressor elements and low-temperature and MeJA-responsive elements. These results suggest that the gene is a novel sugar transporter gene in maize and has important and diverse roles in tolerance to abiotic stresses.

Key words: ZmERD6, Sugar transporter, Promoter, Gene expression, Zea mays

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