DEVELOPMENT OF HIGHLY SENSITIVE PHYTOCHROME DETERMINATION EQUIPMENT BASED ON THE FLUORESCENCE SPECTRA CHANGES CONCOMITANT WITH PHOTOTRANSFORMATION.

本研究基于光敏色素光转化过程中荧光光谱的变化，建立了光敏色素的高灵敏度测定装置，低温下形成的光敏色素光转化中间体在等色点处被光照射后全部转化为Pr。Pr光转化为中间体所引起的荧光光谱变化表明，只有在84 K下观察到的中间体lumi-R才发射出在700 nm处有荧光峰的荧光。新研制的荧光显微计可在-20 ℃下，在666 nm处检测浓度为0.0005的光敏色素溶液，进样量仅为3 μ l。在-140 ℃下进行测定，使666 nm处的吸光度灵敏度提高到0.0001。这个极限比通常的吸收法高出10倍。

Highly sensitive phytochrome determination equipment based on the fluorescence spectra changes concomitant with phototransformation was developed in this research.All phototransformation intermediates formed at low temperature transformed to Pr by light irradiation with isosbestic point. Fluorescence spectra changes which were induced by the phototransformation of Pr to the intermediate suggested that only lumi-R which was the intermediate observed at 84K emitted fluorescence having a peak at 700nm. Other intermediates never emitted fluorescence.Newly developed microfluorometer could detect the phytochrome solution which concentration was 0.0005 in the absorbance at 666nm at -20C.Volume of sample was only 3 ul. Determination which was done at -140C improved the sensitibity to 0.0001 in the absorbance at 666nm. This limit was 10 times higher than the usual absorption method.