甘蓝型油菜BnaWRKY75-like基因cDNA序列的克隆及表达分析  

Cloning and Expression Analysis of Full-length cDNA Sequence of BnaWRKY75-like Gene in Brassica napus

作　　者：尹明智[1] 胡燕[1] 杨胜洁 张瑜 杨露 YIN Mingzhi;HU Yan;YANG Shengjie;ZHANG Yu;YANG Lu(College of Biologyand Agricultural Science&Technology,Zunyi Normal University,Zunyi,Guizhou 563006)

机构地区：[1]遵义师范学院生物与农业科技学院,贵州遵义563006

出　　处：《北方园艺》2020年第9期1-8,共8页Northern Horticulture

基　　金：贵州省科学技术厅、遵义市科学技术局、遵义师范学院联合科技基金资助项目(黔科合LH字[2016]7016号);贵州省高层次创新型人才培养计划资助项目(遵市科合人才[2017]11号);贵州省高等学校创新能力提升计划资助项目(黔教合协同创新字[2013]11)。

摘　　要：WRKY超基因家族能广泛地参与植物的多种生理过程及胁迫反应。该研究以甘蓝型油菜为试验材料,根据转录组测序获得的EST序列设计引物,采用RT-PCR及RACE技术克隆甘蓝型油菜WRKY转录因子基因cDNA全长序列,以期为研究该基因的功能提供参考依据。结果表明:BnaWRKY75-like基因编码序列长444 bp,推导的氨基酸序列编码147个氨基酸残基,在67~124区域含有一个典型的WRKY结构域,其中包含有"WRKYGQK"核心序列,与Brassica rapaWRKY75基因的编码序列相似性为99.77%,氨基酸序列一致。半定量RT-PCR分析表明,低温处理油菜后,BnaWRKY75-like基因的表达增强,表达峰值出现在处理3 h时,其可能在油菜低温胁迫响应中发挥作用。WRKY superfamily was widely involved in many physiological processes and stress responses in plants.In this study,we used Brassica napus as experimental material,designed primers according to EST sequence obtained from transcriptome sequencing,and cloned the full-length cDNA sequence of WRKY transcription factor gene in Brassica napus by RT-PCR and RACE technology,in order to provide reference for the study of the function of the gene.The results showed that the coding sequence of BnaWRKY75-like gene was 444 bp,and the deduced amino acid sequence encoded 147 amino acid residues.There was a typical WRKY domain in the region of 67-124,including the core sequence of’WRKYGQK’.The similarity between the coding sequence of BnaWRKY75-like gene and that of Brassica rapa WRKY75 gene was 99.77%,and the amino acid sequence was the same.Semi quantitative RT-PCR analysis showed that the expression of BnaWRKY75-like gene increased after low temperature treatment,and the peak of expression appeared at 3 hours of treatment,which might play a role in the response to low temperature stress.

关 键 词：甘蓝型油菜 WRKY 转录因子 基因克隆 表达分析 

分 类 号：S565.4[农业科学—作物学]