Department of Biological Sciences, University of Delaware, Newark, Delaware 19716.
Mycologia. 2002 Mar-Apr;94(2):280-9.
The subcellular expression patterns and fluorescence intensities of cytoplasm-targeted, constitutively expressed blue-, cyano-, green-, yellow- and red-fluorescent protein were assessed in a number of transformants of the blast pathogen, Magnaporthe grisea. All transformants grew normally, remained pathogenic on barley, and, except for those expressing blue fluorescent protein, exhibited significant cytoplasmic fluorescence. The exceptionally intense brightness of some strains proved very useful for laser scanning confocal microscope imaging during invasion of host tissues. Acquisition of three-dimensional data sets from intact, individual, pathogen encounter sites in planta were generated during the time course of pathogenesis using non-invasive optical sectioning methods. Confocal and multiphoton microscopy imaging in conjunction with fluorescent protein expression allowed for the real time documentation of fungal colonization within plant cells and tissues with remarkable ease. These methods constitute valuable new tools for the investigation of plant disease.
对稻瘟病菌的一些转化体进行了亚细胞表达模式和荧光强度分析，这些转化体表达的细胞质靶向、组成型表达的蓝色、青色、绿色、黄色和红色荧光蛋白。所有转化体生长正常，在大麦上仍然具有致病性，除了表达蓝色荧光蛋白的转化体外，都表现出明显的细胞质荧光。一些菌株的异常高亮度在激光扫描共聚焦显微镜成像时非常有用，有助于观察入侵宿主组织的过程。在发病过程中，使用非侵入性光学切片方法，从完整的、单个的、病原体接触点获取了三维数据集。共聚焦和多光子显微镜成像以及荧光蛋白表达使得真菌在植物细胞和组织内的定殖能够非常容易地被实时记录下来。这些方法为植物病害的研究提供了有价值的新工具。
