摘要
植物耐盐机制的研究一直是植物抗性研究的焦点。近年来,随着生物学不断发展和新荧光标记技术的运用,胞内钠离子测定逐渐应用于植物盐胁迫研究中。该文论述了以下三方面问题:(1)分别介绍了SBFI、Sodium Green和Coro Na Green三种钠离子荧光指示剂:SBFI是一种双激发波长指示剂,其激发波长是340 nm/380 nm,发射波长是500 nm;Sodium Green和Coro Na Green是单波长指示剂,其激发波长分别是507 nm和492nm,发射波长分别是532 nm和516 nm;(2)比较了酯导入、酸导入、电穿孔和显微注射等几种常见荧光指示剂载入胞内方法的优缺点,重点介绍了一种无损伤低温抑制酯酶法:先将荧光指示剂在缓冲液中低温(4℃)处理2 h,随后回到常温(20℃)在不含荧光指示剂的缓冲液中孵育2 h;(3)阐述了胞内离子浓度计算公式,包括单波长测定公式、双波长比率测定公式。
The mechanism of salt tolerance in plants had been the focus of plant resistance research in the past years.As the development of biology and application of new fluorescent labeling technologies,determination of intra-cellular free sodium had been gradually applied to the study of salt tolerance in plants.This review discussed three points as follows：（1）Introduction of three fluorescent indicators of intracellular free sodium：SBFI,Sodium Green and CoroNa Green.SBFI was a kind of fluorescent indicator for excitation ratio measurements,its emission ratio de-tected at 500 nm when excited at 340/380 nm.Sodium Green and CoroNa Green were fluorescent indicators that lacked a significant shift in emission or excitation wavelength upon binding to Na＋ .Sodium Green and CoroNa Green could be detected at 532 nm and 516 nm when excited at 507 nm and 492 nm respectively;（2）Compared the advan-tages and disadvantages of the protocols of loading the fluorescent indicators into cells,including AM esters of the flu-orescent probes,acid loading,electroporation and microinjection.A non-invasive loading of acetoxymethyl ester under low temperature was introduced：loading the fluorescent indicator into cells by incubating the cells in solution at 4 ℃for 2 h followed by 2 h incubation in the dye-free solution at 20 ℃;（3）The measurement of the internal sodium con-centration in cells was illustrated.The equation for measurement of fluorescence intensity that lacked a significant shift in emission or excitation wavelength was：[Na^ ＋]=K d （F -F min ）/（F max -F ）.Fluorescence intensity （F ）wastargeted fluorescence intensity.F min was appropriate mixtures of low Na＋ ,and F max was appropriate maximum of high Na＋ .The equation for measurement of fluorescence intensity ratio was：[[Na^ ＋]= K dQ（R - R min ）/（R max -R ）.The ratio of fluorescence intensity （R ）was the ration F1/F2 of the fluorescence intensity.F min was the ratio of appropriate mixtures of low Na^＋ ,and F max was the rat
出处
《广西植物》
CAS
CSCD
北大核心
2015年第3期442-446,441,共6页
Guihaia
基金
中国科学院(仪器设备功能开发)技术创新项目(2014年)
关键词
细胞内游离
NA^+
植物
荧光指示剂
低温酯导入
离子浓度计算
intracellular free sodium
plants
fluorescent indicators
ester loading under low temperature
calculating the ion concentration
