响应ABA的甘草bZIP转录因子的基因表达及调控研究  

Gene Expression and Regulation of bZIP Transcription Factors in Response to ABA in Glycyrrhiza uralensis Fisch.

作　　者：王清 武立伟 范潘慧 徐志超 罗红梅[1] 孙伟[3] 王瑀[1] 郑司浩 宋经元[1,5] 姚辉[1,5] WANG Qing;WU Li-wei;FAN Pan-hui;XU Zhi-chao;LUO Hong-mei;SUN Wei;WANG Yu;ZHENG Si-hao;SONG Jing-yuan;YAO Hui(Key Lab of Chinese Medicine Resources Conservation,State Administration of Traditional Chinese Medicine of the People′s Republic of China,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China;College of Life Sciences,Northeast Forestry University,Harbin 150040,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;China National Traditional Chinese Medicine Co.,Ltd.,Beijing 102600,China;Engineering Research Center of Chinese Medicine Resources,Ministry of Education,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所/国家中医药管理局中药资源保护重点研究室,北京100193 [2]东北林业大学生命科学学院,黑龙江哈尔滨150040 [3]中国中医科学院中药研究所,北京100700 [4]中国中药有限公司,北京102600 [5]中药资源教育部工程研究中心,北京100193

出　　处：《中国现代中药》2023年第6期1207-1217,共11页Modern Chinese Medicine

基　　金：国家自然科学基金项目(32070368);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-071)。

摘　　要：目的:研究响应脱落酸(ABA)的甘草碱性亮氨酸拉链(GubZIPs)转录因子的基因表达差异,克隆并分析甘草关键酶基因的启动子序列,为解析bZIP转录因子调控甘草活性成分生物合成提供参考。方法:采用实时荧光定量聚合酶链式反应(qRT-PCR)分析甘草毛状根中GubZIPs转录因子基因在外源ABA处理下的表达模式及在3年生甘草中的空间表达差异;利用TBtools软件从甘草基因组中提取关键酶基因的启动子片段并克隆,利用生物信息学分析启动子上潜在的关键调控元件。结果:甘草GubZIP1、GubZIP5、GubZIP33和GubZIP56基因在不同的ABA处理条件下表达模式不同,其中GubZIP1和GubZIP33的响应最为显著。分析GubZIPs基因在甘草不同组织的表达差异发现,GubZIP1和GubZIP5在根部高水平表达,GubZIP33和GubZIP56在叶中高水平表达。基于甘草基因组克隆获得甘草生物合成途径中关键酶查耳酮异构酶(CHI)、查耳酮合成酶(CHS)、β-香树脂醇合酶(β-AS)基因的启动子序列,分析结果显示,CHI基因的启动子区域有2个G-box和1个A-box顺式作用元件,CHS基因的启动子区域有2个ABRE和1个G-box顺式作用元件,β-AS基因的启动子区域有1个ABRE和1个G-box顺式作用元件。结论:甘草毛状根中GubZIP1、GubZIP5、GubZIP33和GubZIP56基因对ABA均有显著响应,采用50 mg·L^(–1)的ABA处理3 h是研究甘草中ABA相关通路较为适宜的方案。此外甘草的关键酶基因启动子上存在可与bZIP转录因子结合和响应ABA等激素的顺式作用元件;研究结果可为深入解析响应ABA的bZIP转录因子调控甘草活性成分生物合成的分子机制提供参考。Objective:To study the gene expression differences of GubZIPs transcription factors in response to abscisic acid(ABA),clone and analyze the promoter sequences of key enzyme genes of Glycyrrhiza uralensis Fisch,and provide references for analyzing the bZIP transcription factors regulating the biosynthesis of active components in G.uralensis.Methods:Real-time quantitative polymerase chain reaction(qRT-PCR)was adopted to analyze the expression pattern of GubZIPs under exogenous ABA treatment in the hairy root of G.uralensis,and then the spatial expression differences of GubZIPs in triennial G.uralensis was studied.The promoter fragments of G.uralensis key enzymes were obtained and cloned from the G.uralensis genome by the TBtools software,and the potential key regulatory elements on the promoters were analyzed by bioinformatics.Results:The expression patterns of the G.uralensis genes GubZIP1,GubZIP5,GubZIP33,and GubZIP56 vary in different ABA treatment conditions,with the most significant responses of GubZIP1 and GubZIP33.Analysis on the expression differences of GubZIPs in different tissues of G.uralensis shows that GubZIP1 and GubZIP5 are highly expressed in the root,and GubZIP33 and GubZIP56 are highly expressed in the leaves.Based on the G.uralensis genome,the promoter sequences of key enzyme genes CHI,CHS,andβ-AS in the G.uralensis biosynthesis pathway were cloned.The analysis results demonstrate that the promoter region of the CHI gene has two G-box and one A-box cis-acting elements,that of the CHS gene has two ABRE and one G-box cis-acting elements,and that of theβ-AS gene has one ABRE and one G-box cis-acting elements.Conclusion:The GubZIP1,GubZIP5,GubZIP33,and GubZIP56 in hairy roots of G.uralensis exhibit significant responses to ABA.Treatment with 50 mg·L^(–1) of ABA for three hours is a suitable scheme for studying the ABA-related pathways in G.uralensis.In addition,cis-acting elements that can bind to bZIP transcription factors and respond to ABA and other hormones exist in the promoter regions of

关 键 词：甘草 毛状根 亮氨酸拉链转录因子 空间表达 关键酶基因 启动子 

分 类 号：R282.12[医药卫生—中药学]