用同步辐射X-射线荧光分析法研究感病烟草单细胞内微量元素变化

Using Synchrotron Radiation X-ray Fluorescence Spectrometer to Study Trace Element Changes in Single Tobacco Protoplast Infected by TMV

用同步辐射 X-射线荧光分析 (SRXFA)技术分析植物单细胞的微量元素 ,研究病毒侵染植物后植物单原生质体内微量元素的相对变化。用果胶酶和纤维素酶分离烟草叶肉细胞原生质体 ,经 3%戊二醛固定 30min后 ,置于杜乐膜上 ,干燥后用同步辐射 X-射线荧光法分析。 X-射线流强大于 80 m A时 ,停留取谱时间2 0 min,可从烟草单个原生质体中检测到 Cl,Ca,Mn,Fe,Cu,Zn,Cr,Co,Ni,As,Ti和 Ge。流强低于 80 m A时 ,只可检出 Cl,Ca,Fe,Cu,Ti和 Ge。结果表明烟草被烟草花叶病毒感染后 ,Ca,Fe离子的强度下降 ,Cl,Cu,Ge离子变化不显著。

The feasibility of using the energy dispersion synchrotron radiation X ray fluorescence (SRXFA) for analyzing trace elements in tobacco protoplasts was demonstrated. Protoplasts were isolated from tobacco leaves, fixed by 3% glutaraldehyde for 30min, then put on Dular film. After drying, protoplasts were analyzed by SRXFA. Acquisition time lasting 20min, Cl, Ca, Mn, Fe, Cu, Zn, Cr, Co, Ni, As, Ge and Ti were detected in single protoplast when beam intensity was higher than 80mA; while current was lower than 80mA, only Cl, Ca, Fe, Cu, Ge and Ti could be detected. Our data show that, after protoplasts were infected by tobacco mosaic virus, the concentrations of Ca and Fe had a significant decrease compared with health protoplast, and Cl, Cu and Ge did not change.