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选择奥地利境内阿尔卑斯山健康欧洲云杉为对象,研究室内接种蓝变真菌(Ceratocystis polonica)引起的寄主树木韧皮部和木质部解剖学特征的变化,揭示蓝变真菌引起欧洲云杉枯萎的机制。结果表明:接种1周后的4株欧洲云杉的木质部组织内,蓝变区域显著增加,4~6周后蓝变区域不再增加;而在接种无菌琼脂的2株对照欧洲云杉的木质部组织内,没有检测到蓝变区域。采用生物化学分析和组织定位技术,确定接种真菌和无菌琼脂的欧洲云杉木质部区域纤维素酶的分布和活性变化。接种2周后剥皮取样检测,接种真菌的4株欧洲云杉的木质部组织内纤维素酶含量大幅度增加,其等电聚焦电泳显示明显的纤维素酶酶带;而在接种无菌琼脂的欧洲云杉木质部区域,纤维素酶含量分布较少,其等电聚焦电泳显示微弱的纤维素酶酶带。进一步证明蓝变真菌分泌的纤维素酶能利用寄主欧洲云杉木质部的纤维素,蓝变真菌是致死阿尔卑斯山境内欧洲云杉的重要病原菌。
The selection of the healthy European spruce in the Alps in Austria was carried out to study the anatomical changes of the phloem and xylem in the host tree caused by Ceratocystis polonica inoculation, and to reveal the mechanism of the blue-change fungus causing the spruce of European spruce. The results showed that within 1 week after inoculation, the blue-change area increased significantly in 4 European spruce tissues, and the blue-change area did not increase after 4 to 6 weeks. In the two control spruce inoculated with sterile agar, Within the xylem tissue, no blue-shifted area was detected. Biochemical analysis and tissue localization techniques were used to determine the distribution and activity of cellulase in the xylem region of European spruce with inoculation of fungi and sterile agar. Two weeks after inoculation, the skin samples were taken for examination. The cellulase content in the xylem tissues of four European spruce inoculated with fungi increased significantly, and the isoenzymatic electrophoresis showed obvious cellulase bands. In the European Spruce xylem region, the distribution of cellulase less, the isoelectric focusing electrophoresis showed weak cellulase bands. It is further demonstrated that the cellulase secreted by the bluefungal fungi can utilize the cellulose of the xylem of the host European spruce, and that the bluefungal fungi are the important pathogens causing the death of the European spruce in the Alps.