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测定植物胞内游离钠离子的研究进展(PDF)

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2015年03期
页码:
442
栏目:
专论与综述
出版日期:
2015-05-20

文章信息/Info

Title:
Progress of the study on determination of free sodium in plant cells
文章编号:
1000-3142(2015)03-0442-05
作者:
刘瑞娟1 蔡振媛1 车国冬2*
1. 中国科学院高原生物适应与进化重点实验室, 西宁 810001; 2. 中国科学院西北高原生物研究所, 西宁 810001
Author(s):
LIU Rui-Juan1 CAI Zhen-Yuan1 CHE Guo-Dong2*
1. Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China; 2. Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China
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
Keywords:
intracellular free sodium plants fluorescent indicators ester loading under low temperature calculating the ion concentration
分类号:
Q942; Q2-33
DOI:
10.11931/guihaia.gxzw201404034
文献标识码:
A
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CoroNa 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CoroNa Green是单波长指示剂,其激发波长分别是507 nm和492 nm,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。
Abstract:
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 intracellular 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 detected 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 advantages and disadvantages of the protocols of loading the fluorescent indicators into cells,including AM esters of the fluorescent 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 concentration in cells was illustrated. The equation for measurement of fluorescence intensity that lacked a significant shift in emission or excitation wavelength was:[Na+]=Kd(F-Fmin)/(Fmax-F). Fluorescence intensity(F)was targeted fluorescence intensity. Fmin was appropriate mixtures of low Na+,and Fmax was appropriate maximum of high Na+. The equation for measurement of fluorescence intensity ratio was:[[Na+]=KdQ(R-Rmin)/(Rmax-R). The ratio of fluorescence intensity(R)was the ration F1/F2 of the fluorescence intensity. Fmin was the ratio of appropriate mixtures of low Na+,and Fmax was the ratio of appropriate maximum of high Na+.

参考文献/References

Amorino GP,Fox MH. 1995. Intracellular Na+ measurements using sodium green tetraacetate with flow cytometry[J]. Cytometry,21(3):248-256
Bi Y(毕莹), Lu RY(卢芮伊), Zhou X(周欣), et al. 2012. Evaluation of a flow cytometry based intracellular sodium detection assay by CoroNaTM Green(基于CoroNaTM Green检测细胞内Na+浓度的流式细胞术分析方法的建立和评价)[J]. J Tianjin Med Univ(天津医科大学学报),18(4):428-431
Bkaily G, Jacques D, Pothier P. 1999. Use of confocal microscopy to investigate cell structure and function[J]. Meth Enzymol,307:119-135
Black JA,Waxman SG. 1996. Sodium channel expression: a dynamic process in neurons and non-neuronal cells[J]. Dev Neurosci,18:139-152
Brownlee C,Pulsford AL. 1988. Visualization of the cytoplasmic free Ca2+ gradient in Fucus serratus rhizois: correlation with cell ultrastructure and polarity[J]. J Cell Biol,81:249
Bush DS,Jones RL. 1987. Measurement of cytoplasmic calcium in aleurone protoplasts using indo-1 and fura-2[J]. Cell Cal,8(6):455-472
Chen JN(陈介南),Wang Q(王琼),Lu MZ(卢孟柱),et al. 2010. Application of quantum dots in plant molecule fluorescent labeling(量子点在植物分子荧光标记中的应用)[J]. Sci Sil Sin(林业科学),46(10):153-161
Dang L(党磊),Wang DM(王冬梅),Liu J(刘娟),et al. 2002. Immunofluorescence localization of microtubule in wheat mesophyll protoplast and its influence factors(小麦叶片原生质体微管骨架的免疫荧光标记及其影响因素)[J]. J Agric Univ Hebei(河北农业大学学报),25(3):23-27
Despa S,Vecer J,Steels P,et al. 2000. Fluorescence lifetime microscopy of the Na+ indicator sodium green in HeLa cells[J]. Analyt Biochem,281(12):159-175
Ding F(丁烽). 2010. The salt-secretion mechanism of salt glands in the leaves of Limonium Bicolor(二色补血草叶片盐腺泌盐机理的研究)[D]. Jinan(济南):Shandong Normal University(山东师范大学) Duan XG,Yang AF,Gao F, et al. 2007. Heterologous expression of vacuolar H+-PPase enhances the electrochemical gradient across the vacuolar membrane and improves tobacco cell salt tolerance[J]. Protoplasma,232(1-2):87-95
Guo Y(郭祎),Ren ZY(任兆玉),Hou X(侯洵). 2003. Fluorescence methods for analyzing in tracellular second messenger-calcium ion(细胞内第二信使-钙离子荧光测定方法的研究进展)[J]. Laser J(激光杂志),24(10):1-5
Halperin SJ,Lynch JP. 2003. Effects of salinity on cytosolic Na+ and K+ in root hairs of Arabidopsis thaliana: in vivo measurements using the fluorescent dyes SBFI and PBFI[J]. J Exp Bot,54(390): 2 035-2 043
Kader MA,Lindberg S. 2005. Uptake of sodium in protoplasts of salt-sensitive and salt-tolerant cultivars of rice,Oryza sativa L. determined by the fluorescent dye SBFI[J]. J Exp Bot,56(422):3 149-3 158
Kavitha PG,Miller AJ,Mathew MK, et al. 2012. Rice cultivars with differing salt tolerance contain similar cation channels in their root cells[J]. J Exp Bot,63(8):3 289-3 296
Lo CJ,Leake MC,Berry RM. 2006. Fluorescence measurement of intracellular sodium concentration in Single Escherichia colic cells[J]. Biophys J,90(1):357-365
Luo HY(罗红艺),Jing HJ(景红娟). 2002. Sodium, nickel and silicon, new essential elements in nutrients of plant(植物营养中新的必需元素——钠、镍、硅)[J]. J Higher Corr Educ:Nat Sci Ed(高等函授学报·自然科学版),15(3):42-45
Martin VV,Rothe A,Gee KR. 2005. Fluorescent metal ion indicators based on benzoannelated crown systems:a green fluorescent indicator for intracellular sodium ions[J]. Bioorg & Med Chem Lett,15(7):1 851-1 855
Meier SD,Kovalchuk Y,Rose CR. 2006. Properties of the new fluorescent Na+ indicator CoroNa green:comparison with SBFI and confocal Na+ imaging[J]. J Neurosci Meth,155(2):251-259
Minta A,Tsien RY. 1989. Fluorescent indicators for cytosolic sodium[J]. J Chem,264:19 449-19 457
Negulescu PA,Machen TE. 1990,Intracellular ion activities and membrane transport in arietal cells measured with fluorescent dyes[J]. Meth Enzymol,192:38-81
Shang ZL(尚忠林),Wang YF(王永飞),Qian H(钱洪),et al. 2001. The measurement of calcium fluorescence in lily pollen cells(百合花粉细胞中钙离子的荧光测定法)[J]. Plant Physiol J(植物生理学通讯),37(4):319-322
Sun QP(孙清鹏),Wang XJ(王小菁). 2004. The detection of cytosolic free Ca2+ in cells of Arabidopsis thaliana leaves(拟南芥叶细胞游离钙离子的测定)[J]. J Trop & Subtrop Bot(热带亚热带植物学报),12(1):79-82
Sun TE(孙天恩),Zhou P(周平),Ye MW(叶梦炜). 1996. Progress of the study on determination of free calcium in plant cells by fluorescent indicator(荧光指示剂测定植物细胞内游离钙离子的研究进展)[J]. Plant Physiol Comm(植物生理学通讯),32(2):91-99
Wang XL(王晓玲). 2009. Construction of a Salt Gland Specific cDNA Library from Limonium(补血草盐腺细胞特异性cDNA文库构建)[D]. Jinan(济南):Shandong Normal University(山东师范大学) Wang YX(王永祥),Liu B(刘斌). 2006. Development of fluorescent indicators of intracellular free Ca2+(细胞内游离 Ca2+的荧光指示剂研究进展)[J]. J Xi’an Univ Arts & Sci:Nat Sci Ed(西安文理学院学报·自然科学版),9(1):21-24
Xu GH(徐国华),Zhang SL(张绍玲),Zhang CY(张超英),et al. 2003. Changes in free Ca2+ distribution in pollen cell after self-and cross pollination in Pyrus serotina Re hd(梨自花与异花授粉后花粉胞内游离 Ca2+分布的变化)[J]. J Plant Physiol & Mol Biol(植物生理与分子生物学学报),29(2):97-103
Yu XC(余晓丛),Zhang SY(张少英),Na R(娜仁). 2013. Optimization of fluorescence detection conditions of calcium ion in the cell in sugar beet(荧光指示剂检测甜菜胞内钙离子的条件优化)[J]. J Northeast Agric Univ(东北农业大学学报),44(1):61-64
Yuan JH(袁建辉),Yang H(杨慧),Tang H(唐焕),et al. 2011. A cell-based detection of ciguatoxin usingsodium fluorescence probe(钠离子荧光探针用于检测雪卡毒素的细胞毒性)[J]. J Southern Med Univ(南方医科大学学报),31(4):653-655
Zhang JT,Liu H,Sun J, et al. 2012. Arabidopsis fatty acid desaturase FAD2 is required for salt tolerance during seed germination and early seedling growth[J]. PLoS One,7(1):1-12
Zhang WH,Rengel Z,Kuo J. 1998. Determination of intracellular Ca2+ in cells of intact wheat roots:loading of acetoxymethyl ester of Fluo-3 under low temperature[J]. Plant J,15(1):147-151
Zhou P(周平),Sun TE(孙天恩),Ye MW(叶梦炜), et al. 1995. Study on the loading of Ca2+ fluorescent indicator into protoplasts(钙离子荧光指示剂导入植物细胞原生质体的研究)[J]. Wuhan Univ J:Nat Sci Ed(武汉大学学报·自然科学版),41(2):208-212

备注/Memo

备注/Memo:
收稿日期: 2014-05-26修回日期: 2014-07-20
基金项目: 中国科学院(仪器设备功能开发)技术创新项目(2014年)
作者简介: 刘瑞娟(1981-),女,甘肃成县人,硕士,工程师,从事植物分子生物学与细胞生物学新技术应用研究,(E-mail)Rjliu@nwipb.cas.cn。 *通讯作者: 车国冬,硕士,工程师,研究方向为天然药物化学,(E-mail)gdche@nwipb.cas.cn。
更新日期/Last Update: 2015-05-20