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本文应用共聚焦激光扫描显微术(CLSM)对小麦矮腥黑穗病(TCK)和小麦网腥黑穗病菌(TCT)冬孢子的自发荧光特性进行了研究。研究表明:在这两种冬孢子中,它们各自都含有自发荧光物质,而且都能在488nm、543nm这2种波长激发光激发下,分别在相应的510～530nm、580～600nm范围中得到相应发射光,只是在488nm波长激发光激发下,两种冬孢子发射出的光荧光强度最高,荧光光分布均匀,大体形态清楚;但层切扫描发现自发荧光在这两种冬孢子的空间分布存在明显差异:TCK冬孢子的自发荧光物质主要分布在外孢壁和网脊上,而在原生质中分布少;TCT冬孢子的自发荧光物质主要分布在原生质中,孢壁分布少,而网脊上不分布。应用CLSM在实际TCK检测工作中,能克服荧光显微镜存在的检测重复性较差、可靠性小的缺点,提升了TCK检验检疫的技术水平。
In this paper, the autofluorescence characteristics of teliospores TCZ and TCT were studied by confocal laser scanning microscopy (CLSM). It has been found that both of them contain autofluorescent materials, and both of them can be respectively corresponding to 510-530 nm and 580-600 nm under the excitation of 488 nm and 543 nm respectively However, only the excitation light at 488nm excitation, two teliospores emitted the highest fluorescence intensity, uniform fluorescence distribution, the general shape of the clear; but by stratum scan revealed the existence of autofluorescence in the two spatial distribution of teliospores Significant differences: TCF teliospores autofluorescent substances are mainly distributed in the outer spores and ridges, but less distributed in the protoplasm; TCT teliospores autofluorescent substances are mainly distributed in the protoplasm, spore wall less distributed, and the net ridge is not distributed. The application of CLSM in actual TCK detection can overcome the shortcomings of fluorescence microscopy, such as poor repeatability and low reliability, and improve the technical level of TCK inspection and quarantine.