YIN Mingzhi,HU Yan,YANG Shengjie,et al.Cloning and Expression Analysis of Full-length cDNA Sequence of BnaWRKY75-like Gene in Brassica napus[J].Northern Horticulture,2020,44(09):1-8.[doi:10.11937/bfyy.20193740]
甘蓝型油菜BnaWRKY75-like基因cDNA序列的克隆及表达分析
- Title:
- Cloning and Expression Analysis of Full-length cDNA Sequence of BnaWRKY75-like Gene in Brassica napus
- Keywords:
- Brassica napus; WRKY; transcription factors; gene cloning; expression analysis
- 文献标志码:
- A
- 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 rapa [STBX]WRKY75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。
- Abstract:
- 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.
参考文献/References:
[1]EULGEM T,RUSHTON P J,ROBATZEK S,et al.The WRKY superfamily of plant transcription factors[J].Trends Plant Sci,2000,5(5):199-206.[2]XING D H,LAI Z B,ZHENG Z Y,et al.Stress-and pathogen-induced Arabidopsis WRKY48 is a transcriptional activator that represses plant basal defense[J].Mol Plant,2008,1(3):459-470.[3]WU X L,SHIROTO Y,KISHITANI S,et al.Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRK-Y11 under the control of HSP101promoter[J].Plant Cell Reports,2009,28(1):21-30.[4]伍林涛,钟贵买,王健美,等.转录因子AtWRKY28在拟南芥与甘蓝链格孢菌(Alternaria brassicicola)亲和性互作中的功能分析[J].中国农业科技导报,2012,14(1):65-71.[5]邹克琴,汪得凯,胡东维,等.水稻WRKY转录因子OsWRKY71的cDNA分离和表达分析[J].浙江农业学报,2014,26(6):1405-1411.[6]石红梅,战新梅,管世铭,等.牡丹PsWRKY基因的克隆和表达特性分析[J].植物生理学报,2015,51(10):1743-1748.[7]文亦蒂,单贵莲,姜飞,等.柱花草WRKY转录因子在低磷胁迫下的克隆与分析[J].西北植物学报,2019,39(2):226-233.[8]刁玄章,王述彬,刁卫平,等.辣椒CaWRKY8基因克隆及胁迫下的表达分析[J].西北植物学报,2019,39(2):210-217.[9]杨阳,李玥莹,张颖.WRKY转录因子与脱落酸在调控低温中的作用[J].中国农学通报,2017,33(29):31-35.[10]王明芳,李卓夫,王晓楠,等.低温下冬小麦品种间WRKY转录因子的表达特性[J].麦类作物学报,2015,35(2):151-158.[11]ZHANG Y,YU H,YANG X,et al.CsWRKY46,a WRKY transcription factor from cucumber,confers cold resistance in transgenic-plant by regulating a set of cold-stress responsive genes in an ABA-dependent manner[J].Plant Physiology & Biochemistry,2016,108:478-487.[12]QIU Y P,YU D Q.Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis[J].Environmental and Experimental Botany,2009,65(1):35-47.
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备注/Memo
第一作者简介：尹明智（1983-），男，博士，副教授，现主要从事油菜功能基因等研究工作。E-mail:277536002@qq.com.基金项目：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）。收稿日期：2020-01-11