拟南芥吲哚族芥子油苷合成调控基因表达模式的分析  

Expression Pattern Analysis of Regulatory Genes Responsible for Indolic Glucosinolate Biosynthesis in Arabidopsis thaliana

作　　者：盛宇欣 彭疑芳 朱星宇 金峰[1] 孔稳稳[1] 李晶[1] SHENG Yu-Xin;PENG Yi-Fang;ZHU Xing-Yu;JIN Feng;KONG Wen-Wen;LI Jing(College of Life Science, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《农业生物技术学报》2019年第4期606-614,共9页Journal of Agricultural Biotechnology

基　　金：黑龙江省教育厅科学技术研究项目(No.12531003)

摘　　要：吲哚族芥子油苷(indolic glucosinolate, IGS)在植物对病原菌的防御反应中具有重要作用,MYB(myeloblastosis) 34、MYB51和MYB122是调控IGS合成的重要转录因子。本研究拟系统研究3个转录因子基因的时空表达模式及其对激素的响应,为深入探讨吲哚族芥子油苷的抗病机制提供数据参考。首先以拟南芥(Arabidopsis thaliana)基因组DNA为模板,克隆MYB34、MYB51和MYB122基因的启动子序列,构建由启动子驱动报告基因GUS (β-glucuronidase编码基因)的植物表达载体,转入拟南芥后获得PMYB34::GUS、PMYB51::GUS和PMYB122::GUS转基因株系。通过GUS活性信号检测,对基因的空间表达模式及激素响应性进行分析。基因表达分析显示,上述3个基因的时空表达模式呈现一定的相似性,即在生长发育的各个时期及各种不同组织中均有表达:在幼苗的子叶、胚轴和胚根及成熟植株的根、茎、叶、花和角果的维管束等处均有表达;但是基因表达也存在明显差异,其中MYB51基因在地上部分的表达水平显著高于根,MYB34和MYB122则主要在根中表达。激素响应性分析显示,MYB51受到水杨酸强烈诱导,MYB34则对茉莉酸甲酯最为敏感,处理后表达水平显著提高;与MYB51和MYB34相比,MYB122对各种激素的响应相对不敏感,在本研究的实验条件下,几种激素处理后其基因表达水平未发生大幅度变化。上述研究结果提示,IGS代谢在植物不同部位可能受到不同信号途径调控。本研究初步揭示了MYB34、MYB51和MYB122的基因表达特点和激素响应特性,为深入揭示IGS合成调控机制提供了基础资料。Indolic glucosinolates(IGS) play a crucial role in plant pathogen defense responses, MYB(myeloblastosis) 34, MYB51 and MYB122 are 3 important transcription factors that regulate indolic glucosinolate biosynthesis. This study systematically studied the temporal and spatial expression patterns of 3 transcription factor genes and their responses to hormones, which could provide data for further study on the disease resistance mechanism of indolic glucosinolates. Using Arabidopsis thaliana genomic DNA as a template, the promoter sequences of MYB34, MYB51 and MYB122 genes were amplified, plant expression vectors with GUS(β-glucuronidase coding gene) expression driven by the promoters of MYB34, MYB51, and MYB122, respectively were constructed and transferred into Arabidopsis thaliana. PMYB34::GUS, PMYB51::GUS and PMYB122:: GUS transgenic lines were confirmed. Through the detection of GUS activity, the temporal and spatial expression patterns of 3 genes and the response to several hormones were analyzed. Gene expression analysis showed that the temporal and spatial expression patterns of MYB34, MYB51 and MYB122 genes were similar. All the 3 genes generally expressed in various stages of growth and development and in various tissues, in the cotyledons, hypocotyls and radicles of seedlings, and the vascular bundles of roots,stems, leaves, flowers, and pods of mature plants. However, the 3 genes showed their own specific expression pattern. The expression of MYB51 performed higher in ground tissue than in root tissue, while the MYB34 and MYB122 were mainly expressed in the roots. Hormone responsiveness analysis showed that MYB51 was induced by salicylic acid(SA) and MYB34 was induced by methyl jasmonate(MeJA). Compared with MYB34 and MYB51, MYB122 was not sensitive to hormone stimulation. Under the experimental conditions of this study, the expression level of MYB122 did not change significantly after several hormone treatments. The above research results suggested that the metabolism of indolic glucosinolates might be re

关 键 词：拟南芥 吲哚族芥子油苷(IGS) MYB (myeloblastosis)转录因子 时空表达模式 激素响应性 

分 类 号：S184[农业科学—农业基础科学]