摘要
克隆蒙古沙冬青钾离子通道AmGORK的启动子区域,探明在干旱胁迫下它对保卫细胞运动的调控机制,为深入研究该基因在水分及养分利用、光合作用、抗旱等过程的功能奠定基础。以蒙古沙冬青保卫细胞外向钾离子通道基因AmGORK编码序列为基础,通过染色体步移技术克隆该基因的启动子序列,并对其进行生物信息学分析,构建pCambia1301-AmGORK∷GUS双元载体转化野生型拟南芥,对T3转基因拟南芥各组织进行GUS活性染色,转基因植株经PEG或ABA处理后,分析AmGORK启动子的表达情况。结果表明,获得AmGORK上游1723 bp序列,该区域含有多个响应干旱、ABA、光照等的顺式调控元件,说明AmGORK可能参与干旱响应过程。经GUS活性染色,发现AmGORK在根中柱、叶柄、叶脉、叶片保卫细胞、花萼均有表达,表明AmGORK的表达具有组织特异性。定量分析发现PEG或ABA处理后的转基因植株中GUS分别上调3.2倍和4.1倍。AmGORK表达可能受干旱诱导。
To investigate its regulation mechanism on guard cell movement under drought stress,the promoter region of potassium channel AmGORK in Ammopiptanthus mongolicus was cloned so as to lay a foundation for further studying the functions of AmGORK in water and nutrient utilization,photosynthesis,drought resistance and other processes.Based on the coding sequence of the outward potassium channel gene AmGORK in the guard cells of A.mongolicus,the promoter sequence of the gene was cloned by chromosome walking technique,and its bioinformatics analysis was further conducted.The pCambia1301-AmGORK∷GUS binary vector was constructed and transformed into the wild-type A.thaliana.The tissues of T3 in the transgenic A.thaliana were stained for analyzing GUS activity.After being treated with PEG or ABA,the transgenic plants were analyzed for the expression of AmGORK promoter.The results showed that the 1723 bp upstream sequence of AmGORK was obtained,and contained several ci regulatory elements in response to drought,ABA and light,indicating that AmGORK may be involved in the drought response process.GUS activity staining showed that AmGORK was expressed in the root stele,petiole,vein,guard cell of a leaf,and calyx,indicating that the expression of AmGORK was tissue-specific.According to quantitative analysis,GUS was up-regulated by 3.2-fold and 4.1-fold in the PEG-or ABA-treated transgenic plants,respectively.Consequently,the expression of AmGORK may be induced by drought.
作者
李俊林
张焕朝
聂文婧
张海洋
王向誉
郭洪恩
韩蕾
LI Jun-lin;ZHANG Huan-chao;NIE Wen-jing;ZHANG Hai-yang;WANG Xiang-yu;GUO Hong-en;HAN Lei(Shandong Institute of Sericulture,Yantai 264002;Nanjing Forestry University,Nanjing 210037;Lu Dong University,Yantai 264025)
出处
《生物技术通报》
CAS
CSCD
北大核心
2021年第10期9-16,共8页
Biotechnology Bulletin
基金
国家自然科学基金项目(31601819)
山东省农业科学院农业科技创新工程项目(CXGC2016B10)
山东省“渤海粮仓”科技示范工程升级版项目(2019BHLC005)
山东省自然科学青年基金项目(ZR2020QC062)。
