茶树荧光性绿斑病叶黄绿色自发荧光的显微分析

茶树荧光性绿斑病是一种茶树成叶生理性病害,病叶上的绿色病斑可见自发的黄绿色荧光。为了以黄、绿色自发荧光为探针了解病害的发生规律,同时为黄色、绿色荧光物质的分离鉴定提供光谱学依据,应用荧光显微技术、显微荧光光谱成像技术、激光扫描共聚焦显微技术、流式细胞术研究了茶树荧光性绿斑病叶中黄、绿色自发荧光的显微观察条件、发射光谱和发光部位。结果表明:在不同激发光照射下,茶树荧光性绿斑病叶能发射多种自发荧光,其中黄色、绿色荧光显微观察的最佳条件为:蓝光激发,彩色和绿色单色光模式记录图像;在绿光、黄光范围有三个荧光峰,其波长分别为515nm、535nm和585nm;最初发出绿色、黄色荧光的病变部位是维管束鞘细胞,随后出现在部分海绵细胞、栅栏细胞和表皮细胞中;黄色、绿色荧光在细胞中的发光位置定位于液泡。

The Microscopic Analysis of Yellow-Greenish Spontaneous Fluorescence in the Tea Leaves of Fluorescent Green Spot Disease

The fluorescent green spot disease is a physiological disease of tea mature leaves. The green spots on the leaf back emit yellow-greenish fluorescence. In order to investigate the rule of the disease occurrence by using spontaneous fluorescence as a probe and the spectra of yellow-greenish fluorescent compounds for their isolation and identification, the fluorescence microscopy, microscopic fluorescence spectral imaging, laser scanning confocal microscopy and flow cytometry were used in this investigation. This experiment covered the condition of microscopic observation, emission spectral characters of yellow-greenish spontaneous fluorescence and their localization in the diseased tea leaves. Results showed that the diseased leaves could emit several kinds of spontaneous fluorescence by irradiating different lights, and the optimal microscopic condition for observing the yellow-greenish fluorescence was that the diseased leaves were excited by blue light and images recorded in multicolor and green color mode, in which there were three fluorescence peaks at 515 nm, 535 nm, and 585 nm within the light range of yellow and green. The yellow-greenish fluorescence were firstly emitted in the sheath cell ofvascular bundle and then in the barrier cells, sponge cells and the epidermis cells. And the location of fluorescence was the vacuole of the diseased cells.