Crops ›› 2017, Vol. 33 ›› Issue (5): 38-42.doi: 10.16035/j.issn.1001-7283.2017.05.007

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Expression and Bioinformatics Analysis of CsWRKY23 Gene in Cucumber

Zhang Ying1,Liu Pengyu2,Bai Xue1,Yang Yang1,Li Yueying1   

  1. 1College of Life Science,Shenyang Normal University,Shenyang 110034,Liaoning,China
    2Shenyang Institute of Food Control,Shenyang 110136,Liaoning,China
  • Received:2017-07-03 Revised:2017-08-25 Online:2017-10-15 Published:2018-08-26
  • Contact: Yueying Li

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Abstract:

WRKY transcription factor is a great gene family that exists in higher plants and widely involved in plant biotic stress and abiotic stress. The cDNA of CsWRKY23 was cloned by RT-PCR, and sequence motif, gene structure and phylogenetic tree were done. The gene expression of CsWRKY23 were detected by qRT-PCR. The results showed that the length of CsWRKY23 open reading frame was 1 431bp, containing 5 exons and 4 introns; encoding 476 amino acids, and there were 2 WRKY motifs, protein molecular weight was 52.2kDa and its iso-electric point was 6.43; phylogenetic analysis showed CsWRKY23 and AtWRKY33 were homologous. Furthermore, qRT-PCR results showed that CsWRKY23 respond to low temperature and up-regulated in leaf and root. In addition, this study provided useful information for further studying this gene and its function under the abiotic stress.

Key words: Cucumber, WRKY, Bioinformatics, Quantitive real time PCR

Fig.1

Analysis of CsWRKY23 structure (A) and prediction of WRKY domain (B)"

Fig.2

Analysis of hydrophobic or hydrophilic property of CsWRKY23"

Fig.3

Prediction of three-dimensional structure of CsWRKY23"

Table 1

Prediction of CsWRKY23 cis-acting elements"

序号Code 元件Element 位置Location 序列Sequence 功能Function
1 WRKY71OS 47 TGAC WRKY转录因子的结合位点
2 TATABOXOSPAL 74 TATTTAA 转录起始位点上游通用转录因子结合的主要的启动原件
3 WRKY71OS 98 TGAC WRKY转录因子的结合位点
4 CAATBOX1 166 CAAT 增强结合蛋白因子
5 CBFHV 225 RYCGAC 响应低温的CBF转录因子,AP2结构域的结合位点
6 MYBCORE 530 CNGTTR MYB转录因子结合位点
7 WRKY71OS 584 TGAC WRKY转录因子的结合位点
8 WRKY71OS 1386 TGAC WRKY转录因子的结合位点
9 ABREATCONSENSUS 1668 YACGTGGC ABA响应过程中的顺势作用原件
10 WRKY71OS 1721 TGAC WRKY转录因子的结合位点
11 WRKY71OS 1749 TGAC WRKY转录因子的结合位点
12 WRKY71OS 1977 TGAC WRKY转录因子的结合位点

Fig.4

Phylogenetic analysis of CsWRKY23"

Table 2

Primers used for PCR assays"

基因Gene 登录号Accession number 引物序列Primer sequence 用途Utility
CsWRKY23 GU984022 F:ATGAGTAACATAAACCAAAC R:CTATGATAGAAATGAGTTG cDNA克隆全长引物
CsWRKY23 GU984022 F:CACTACCTACGAAGGCAAAC R:TGGCTGGTGACACTGGAT qRT-PCR引物
CsActin AAZ74666 F:TCCACGAGACTACCTACAACTC R:CGGAATGGTGAAGGCTGGAT qRT-PCR引物

Fig.5

RT-PCR product of CsWRKY23"

Fig.6

Expression of CsWRKY23 of leaves and roots of cucumber at low temperature"

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