药学学报, 2021, 56(11): 3184-3189
冯飞雪, 吕瑞华, 李依民, 张明英, 王凯, 肇玉博, 高静, 彭亮, 张岗. 北柴胡转录因子BcWRKY70基因鉴定与表达分析[J]. 药学学报, 2021, 56(11): 3184-3189.
FENG Fei-xue, LÜ Rui-hua, LI Yi-min, ZHANG Ming-ying, WANG Kai, ZHAO Yu-bo, GAO Jing, PENG Liang, ZHANG Gang. Characterization and expression analysis of the transcription factor BcWRKY70 in Bupleurum chinense DC.[J]. Acta Pharmaceutica Sinica, 2021, 56(11): 3184-3189.

冯飞雪1, 吕瑞华2, 李依民3,4*, 张明英3, 王凯1, 肇玉博1, 高静3, 彭亮3, 张岗3,4*
1. 陕西中医药大学附属医院, 陕西 咸阳 712000;
2. 陕西中医药大学医学技术学院, 陕西 咸阳 712046;
3. 陕西中医药大学药学院/陕西省秦岭中草药应用开发工程技术研究中心, 陕西 咸阳 712046;
4. 陕西中医药大学省部共建特色秦药资源研究开发国家重点实验室 (培育), 陕西 咸阳 712083
WRKY是植物特有的一类转录因子,参与植物响应多种环境因子及生物胁迫的调控过程。为探究北柴胡WRKY家族成员WRKY70的基因功能,本研究从北柴胡转录组数据库中克隆到BcWRKY70基因开放阅读框 (ORF),进行编码氨基酸序列及DNA启动子区域生物信息学分析,并用实时荧光定量PCR检测BcWRKY70在不同组织部位与不同激素处理下的表达模式。结果表明,北柴胡BcWRKY70基因ORF长948 bp,编码316个氨基酸,具有典型的WRKY结构域,不含信号肽,预测定位在细胞核中;系统进化分析显示BcWRKY70与棉花GhWRKY70亲缘关系最近。BcWRKY70基因启动子区包含响应干旱、茉莉酸甲酯、脱落酸等环境或激素因子的顺式元件。qRT-PCR结果显示BcWRKY70在北柴胡根中表达量最高,且能快速应答茉莉酸甲酯和脱落酸的诱导。这些结果为下一步研究BcWRKY70在北柴胡抗逆中的分子作用机制提供科学依据。
关键词:    北柴胡      WRKY转录因子      克隆      表达模式      启动子区     
Characterization and expression analysis of the transcription factor BcWRKY70 in Bupleurum chinense DC.
FENG Fei-xue1, LÜ Rui-hua2, LI Yi-min3,4*, ZHANG Ming-ying3, WANG Kai1, ZHAO Yu-bo1, GAO Jing3, PENG Liang3, ZHANG Gang3,4*
1. Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712000, China;
2. College of Medical Technology, Shaanxi University of Chinese Medicine, Xianyang 712046, China;
3. College of Pharmacy and Shaanxi Qinling Application Development and Engineering Center of Chinese Herbal Medicine, Shaanxi University of Chinese Medicine, Xianyang 712046, China;
4. State Key Laboratory of Research and Development of Characteristic Qin Medicine Resources (Cultivation), Shaanxi University of Chinese Medicine, Xianyang 712083, China
The plant-specific WRKY transcription factor family is involved in the regulation of the response of plants to various environmental factors and biological stress. To study the role of the BcWRKY70 gene in Bupleurum chinense DC., we cloned the open reading frame (ORF) sequence of the BcWRKY70 gene based on the transcriptome database. Bioinformatics analysis of BcWRKY70 and promoter region analysis was carried out, and real-time qRT-PCR was used to detect the expression pattern of BcWRKY70 in different tissues and under the different hormone treatments. The results show that the BcWRKY70 ORF is 948 bp in length, encoding 316 amino acids with a typical WRKY domain. Phylogenetic analysis showed that BcWRKY70 is closely related to GhWRKY70. Its promoter region contains cis-elements that presumably respond to environmental factors such as drought, methyl jasmonic acid (MeJA), and abscisic acid (ABA). qRT-PCR analyses showed that BcWRKY70 is most highly expressed in roots and can quickly respond to MeJA and ABA induction. This study provides a basis for further research on the molecular mechanism of BcWRKY70 in B. chinense DC. resistance.
Key words:    Bupleurum chinense DC.    WRKY transcription factor    cloning    expression pattern    promoter region   
收稿日期: 2021-07-05
DOI: 10.16438/j.0513-4870.2021-0987
基金项目: 陕西省自然科学基础研究计划(2019JQ-867);陕西中医药大学博士科研启动经费(104080001);陕西中医药大学学科创新团队项目(2019-QN01);咸阳市中青年科技领军人才项目.
通讯作者: 李依民,Tel/Fax:86-29-38185165,E-mail:2051058@sntcm.edu.cn;张岗,E-mail:jay_gumling2003@aliyun.com
Email: 2051058@sntcm.edu.cn;jay_gumling2003@aliyun.com
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