利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响

本研究使用ATP特异性荧光共振能量转移(Fluorescence resonance energy transfer，FRET)为基础的荧光蛋白传感器(Ateam1.03-nD/nA)，分析了4种外源信号分子(细胞外ATP、Ca²⁺、H₂O₂和NO)对拟南芥(Arabidopsis thaliana(L.)Heynh.)幼苗叶绿体和细胞质中ATP水平的影响。结果显示，细胞质ATP水平整体高于叶绿体，在4种不同浓度的信号分子处理下，叶绿体Ateam1.03-nD/nA的FRET比值仅在1.2 ~ 1.8波动；细胞质Ateam1.03-nD/nA 的FRET比值仅在2.2 ～ 3.0之间波动，未产生显著变化。结果表明在以上外源信号分子的作用下，植物细胞质和叶绿体ATP均维持在较为稳定的水平。

Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer

We used an adenosine triphosphate (ATP) fluorescence protein-sensor (Ateam1.03-nD/nA) based on fluorescence resonance energy transfer (FRET) to analyze the effects of extracellular ATP, Ca²⁺, H₂O₂, and NO on ATP levels in the cytoplasm and chloroplasts of Arabidopsis thaliana seedlings. Results showed that ATP levels were higher in the cytoplasm than in the chloroplasts. Under treatment with different concentrations of the signal molecules, the FRET ratio of plastid-localized Ateam 1.03-nD/nA fluctuated non-significantly between 1.2 and 1.8. The FRET ratio of cytosol-localized Ateam1.03-nD/nA fluctuated non-significantly between 2.2 and 3.0. These results indicate that both cytoplasmic ATP and chloroplast ATP maintained relatively stable levels upon stimulation by signal molecules at different intensities.