大豆GmGolS基因高温胁迫应答及启动子活性分析  

Response of Soybean GmGolS Gene to Heat Stress and Promoter Activity Analysis

作　　者：张军 翟莹[2] 邱爽 尹珺伊 张艳 金振华 张勇[3] 王丽坤 ZHANG Jun;ZHAI Ying;QIU Shuang;YIN Jun-yi;ZHANG Yan;JIN Zhen-hua;ZHANG Yong;WANGLi-kun(Branch of Animal Husbandry and Veterinary,Heilongjiang Academy of Agricultural Sciences,Qiqihar 161005,China;College of Life Science and Agro-Forestry,Qiqihar University,Qiqihar 161006,China;Keshan Branch,Heilongjiang Academy of Agricultural Sciences,Qiqihar 161606,China)

机构地区：[1]黑龙江省农业科学院畜牧兽医分院,黑龙江齐齐哈尔161005 [2]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [3]黑龙江省农业科学院克山分院,黑龙江齐齐哈尔161606

出　　处：《大豆科学》2023年第2期188-193,共6页Soybean Science

基　　金：齐齐哈尔市科技计划创新激励项目(CNYGG-2022021);黑龙江省省属高等学校基本科研业务费科研项目(145109506)。

摘　　要：肌醇半乳糖苷合成酶(GolS)是棉子糖系列寡糖(RFOs)生物合成途径中的关键酶。为探究大豆GmGolS基因的高温胁迫调控机制,通过实时荧光定量PCR检测GmGolS在高温胁迫下的表达量,通过PCR从大豆基因组DNA中扩增GmGolS基因启动子序列,将其连接到植物表达载体pCAMBIA1301上并转化烟草,通过GUS组织化学染色和实时荧光定量PCR检测GmGolS启动子的高温诱导启动活性。结果表明:高温胁迫可以强烈诱导GmGolS的表达。1 739 bp的GmGolS启动子序列中含1个生长素响应元件、1个干旱诱导的MYB转录因子结合位点、2个厌氧诱导元件、1个防御和胁迫响应元件、1个脱落酸响应元件、1个茉莉酸响应元件和1个缺氧诱导元件。成功获得3棵GmGolSP转基因烟草植株。GmGolS启动子的活性可以被高温胁迫诱导。Galactinol synthase(GolS) is a key enzyme in the biosynthesis pathway of raffinose family oligosaccharides(RFOs). In order to explore the regulatory mechanism of soybean GmGolS under heat stress, we used real-time fluorescence quantitative PCR to detect GmGolS expression under heat stress, amplified GmGolS promoter sequence by PCR from soybean genomic DNA, constructed GmGolS promoter into plant expression vector pCAMBIA1301 and transformed into tobacco. GUS histochemical staining and real-time fluorescence quantitative PCR were used to detect the heat-induced activation activity of GmGolS promoter. The results showed that the expression of GmGolS was significantly induced by heat stress. The 1 739 bp GmGolS promoter sequence contained one auxin response element, one drought-induced MYB transcription factor binding site, two anaerobic-induced elements, one defense and stress response element, one abscisic acid response element, one jasmonic acid response element and one anoxic-induced element. We obtained three GmGolS promoter transgenic tobacco plants successfully. The activation activity of GmGolS promoter could be induced by heat stress.

关 键 词：大豆 GmGolS 诱导型启动子 肌醇半乳糖苷合成酶 高温胁迫 

分 类 号：S565.1[农业科学—作物学]