大豆生育期基因E1和E2的启动子克隆及其表达模式分析  

Cloning of the Promoters and Analysis of Expression Patterns of Maturity Genes E1 and E2 in Soybean

作　　者：刘路平 胡雪洁 祁金 陈强[1] 刘智 赵田湉 史晓蕾[1] 刘兵强[1] 孟庆民[1] 张孟臣[1] 韩天富[2] 杨春燕[1] LIU LuPing;HU XueJie;QI Jin;CHEN Qiang;LIU Zhi;ZHAO TianTian;SHI XiaoLei;LIU BingQiang;MENG QingMin;ZHANG MengChen;HAN TianFu;YANG ChunYan(Institute of Cereal and Oil Crops,Hebei Academy of Agriculture and Forestry Sciences/Hebei Laboratory of Crop Genetis and Breeding/Huang-Huai-Hai Key Laboratory of Biology and Genetic Improvement of Soybean,Ministry of Agriculture and Rural Affairs,Shijiazhuang 050035;Institute of Crop Sciences,Chinese Academy of Agricultural Sciences/Key Laboratory of Soybean Biology(Beijing),Ministry of Agriculture and Rural Affairs,Beijing 100081)

机构地区：[1]河北省农林科学院粮油作物研究所/河北省作物遗传育种实验室/农业农村部黄淮海大豆生物学与遗传育种重点实验室,石家庄050035 [2]中国农业科学院作物科学研究所/农业农村部北京大豆生物学重点实验室,北京100081

出　　处：《中国农业科学》2025年第5期840-850,共11页Scientia Agricultura Sinica

基　　金：河北省农林科学院科技创新人才队伍建设项目(C22R0303);国家大豆产业技术体系(CARS-04-PS09)。

摘　　要：【目的】生育期是衡量大豆生态适应性的重要指标,也是关乎其产量形成的重要性状。研究大豆生育期主效基因E1和E2启动子及其表达模式,为生育期基因功能研究及其分子调控网络的解析提供依据,为大豆品种适应性改良及产量提高奠定基础。【方法】通过启动子顺式元件分析网站PlantCARE分析生育期基因E1和E2的启动子序列,检测其中包含的重要调控元件。克隆E1和E2的启动子,构建GUS表达载体,并进行拟南芥遗传转化,检测转基因植株不同组织器官的GUS活性。在弱光和强光条件下,比较长日照和短日照下E1和E2表达量的差异。在不同生育期组大豆品种中检测E1和E2表达量,分析其表达量与大豆品种生育期的相关性。【结果】E1和E2的启动子序列均包含AE-box、Box4和G-box等多个光反应元件,另外,E1启动子区还包含生长素、脱落酸等响应元件,E2启动子区包含低温、干旱等响应元件及分生组织表达相关元件。转基因拟南芥GUS活性检测发现,E1启动子在植株整个生长期各器官均有较强转录活性,E2启动子在幼苗下胚轴、叶片和根的维管组织中具有较强转录活性。在弱光和强光条件下,E1表达量均表现为长日照高于短日照。在弱光条件下,E2表达量表现为短日照高于长日照;在强光条件下,E2表达量表现为长日照高于短日照。随不同大豆品种生育期的延长,E1表达量呈现逐渐升高趋势,E2表达量无规律性变化。【结论】E1的启动子是一个广泛表达的启动子,该基因的表达量显著受光周期调控,与大豆品种生育期有明显相关性;E2的启动子在各器官维管组织有较强表达,在强光和弱光条件下,该基因受光周期调控模式存在差异,其表达量与大豆生育期无明显相关性。【Objective】Maturity time is an essential phenotypic measure of ecological adaptability of soybean and an important trait related to its yield formation.The study of promoters and expression patterns of major maturity genes E1 and E2 would provide basis for the study of gene function and molecular regulatory network of maturity time and lay foundation for adaptability improvement and yield increase in soybean.【Method】The promoter sequences of major maturity genes E1 and E2 were analyzed through the promoter cis-element analysis website PlantCARE,and the important regulatory elements were detected.The promoters of E1 and E2 were cloned,the GUS vectors were constructed,and transformation of Arabidopsis was performed to detect GUS activity in different tissues and organs of transgenic plants.Under low light and strong light conditions,the expression levels of E1 and E2 were compared between long day and short day conditions.The expression levels of E1 and E2 were detected in soybean varieties of different maturity groups,which is for the analysis of correlation between expression levels and maturity time of soybean varieties.【Result】Both E1 and E2 promoters contained multiple photoresponsive elements such as AE-box,Box4 and G-box,E1 promoter also contained auxin-response,abolic acid-response elements,and E2 promoter also contained low temperature-response,drought-response elements and meristem expression elements.In GUS activity detection of transgenic Arabidopsis,E1 promoter had strong transcriptional activity in all organs of the plant,and transcriptional activity of E2 promoter in fibrovascular tissues of seedling hypocotyl,leaf and root was relatively strong.Under both low light and strong light conditions,the expression level of E1 was significantly higher in long day than in short day.Under low light conditions,the expression level of E2 was higher in short day than in long day.Under strong light conditions,the expression level of E2 was higher in long day than in short day.With the increase of matu

关 键 词：大豆 生育期 基因E1 基因E2 启动子 表达模式 

分 类 号：R73[医药卫生—肿瘤]