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浙江大学学报（农业与生命科学版）
  2012, Vol. 38 Issue (3): 243-249    DOI: 10.3785/j.issn.1008-9209.2012.03.002
Biological sciences & biotechnology     
Cloning and expression analysis of a transcription factor gene BoWRKY3 from Brassica oleracea var.  italica.
CHEN Beibei1, JIANG Ming1, MIAO Lixiang2, LI Wenping1
1. College of Life Science, Taizhou University, Linhai, Zhejiang 317000, China; 2. Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
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Abstract  A WRKY transcription factor gene, designated BoWRKY3 (accession number: JN120758), was amplified from leaves of Brassica oleracea var.  italica, and the sequence analysis was  performed. The expression patterns  of BoWRKY3 in leaves before and after downy mildew challenge were  analyzed by reverse transcriptionpolymerase chain reaction (RTPCR). Sequencing results indicated that the genomic DNA was 1136 bp in length with 3 introns of 81, 101 and 96 bp, respectively. The complete coding sequence was 858 bp encoding 285 amino acids, including a conserved WRKYGQK domain and a CX5CX23HX1H zinc finger structure. RTPCR analysis showed that the highest expression levels of BoWRKY3 were observed at 636 h induced by Hyaloperonospora parasitica, suggesting that its function was possibly related with downy mildew resistance. Sequence alignment results showed that there were very little differences between BoWRKY3 and homologous  genes of Cruciferae plants, however, major differences were observed between BoWRKY3 and those of Gramineae plants, revealing their large genetic distance.

Published: 24 May 2012
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CHEN Beibei
JIANG Ming
MIAO Lixiang
LI Wenping
Cite this article:

CHEN Beibei, JIANG Ming, MIAO Lixiang, LI Wenping. Cloning and expression analysis of a transcription factor gene BoWRKY3 from Brassica oleracea var.  italica.. , 2012, 38(3): 243-249.

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http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2012.03.002     OR     http://www.zjujournals.com/agr/Y2012/V38/I3/243


青花菜转录因子基因BoWRKY3的克隆与表达分析

以青花菜为材料，从叶片中分离WRKY转录因子基因BoWRKY3（登录号：JN120758），并进行序列分析；利用逆转录聚合酶链式反应（RTPCR）对霜霉菌侵染前后叶片BoWRKY3的表达模式进行分析。测序结果表明：BoWRKY3的基因组DNA全长为1136 bp，有3个内含子，长度分别为81、101和96 bp；编码区全长为858 bp，编码285个氨基酸，具WRKYGQK保守区和CX5CX23HX1H锌指结构。RTPCR结果表明，BoWRKY3的表达受霜霉菌诱导，表达量在接种6~36 h时最高，暗示该基因与青花菜霜霉病抗性相关。序列比对结果表明，BoWRKY3与十字花科同源基因的差异最小，而与禾本科作物的序列差异最大，关系最远。
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