荧光指示剂检测甜菜胞内钙离子的条件优化  

Optimization of fluorescence detection conditions of calcium ion in the cell in sugar beet

作　　者：余晓丛[1] 张少英[1] 娜仁[1] 

机构地区：[1]内蒙古农业大学农学院,呼和浩特010018

出　　处：《东北农业大学学报》2013年第1期61-64,F0002,共5页Journal of Northeast Agricultural University

基　　金：国家自然科学基金(30760125)

摘　　要：胞内钙离子信号做为第二信使参与植物抗病信号转导途径。以甜菜叶片为试验材料,用孵育方法将Ca2+荧光探针Fluo-3/AM载入甜菜细胞中,并结合激光共聚焦扫描显微技术,研究甜菜叶片胞质游离Ca2+的测定方法。结果表明,采用20μmol·L-1的钙离子荧光指示剂Fluo-3/AM装载顺次进行低温4℃孵育120 min,25℃孵育120 min,可获得较为理想的甜菜叶片胞质游离Ca2+染色结果。该方法可用于检测BNYVV侵染甜菜叶片胞质游离钙离子的变化,为研究甜菜BNYVV入侵甜菜后胞内钙信号作用提供理论研究基础。Intracellular calcium signal was a second signal messenger involved in signal transduction pathway of plant resistance to disease. In this study, leaves of sugar beet were used as materials, Ca2. fluorescent probes were embedded into the leaf cell. Meanwhile, methods of measuring cytoplasm free calcium were investigated. The results showed that optimal results of staining cytoplasm free calcium could be obtained by embedding 20 pmol-L-1 fluorescent probe Fluo-3/AM into the leaves followed by incubating 120 min at 4 ℃and 120 min at 25℃. This method can be used to detect change of cytoplasm free calcium in sugar beet leaves after being infected by BNYVV, which provided the theoretical basis for investigating functions of intracellular calcium in signal pathway of sugar beet resistance to BNYVV.

关 键 词：甜菜 胞内Ca2+ 荧光指示剂 激光扫描共聚焦显微技术 

分 类 号：S566.3[农业科学—作物学]