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稻瘟病菌(Magnaporthe oryzae)是异宗配合的子囊菌,但至目前,关于其有性世代产生过程和结构的研究相对较少。本研究利用两个稻瘟病菌菌株Guy-11与2539在多种培养基上进行杂交试验,观察有性世代产生情况。结果表明,两菌株在所有参试培养基上杂交后均能产生子囊壳,但子囊壳的数量、产生速度和成熟度各不相同,以燕麦培养基为最佳。为了进一步观察有性世代的结构,我们采用卡氏白和尼罗红对子囊和子囊孢子进行染色和荧光观察。荧光显微镜下,子囊和子囊孢子的细胞壁均能被卡氏白染成清晰的亮蓝色,细胞结构清晰可辨。成熟的子囊壳内可产生大量的子囊,子囊中含有8个子囊孢子,子囊孢子通常含有4个细胞。同时,子囊孢子能够被尼罗红染成橘红色,表明子囊孢子中储藏大量的脂肪类物质。本研究提供了一种有效的观察稻瘟病菌有性世代结构的荧光染色方法,为后续的研究奠定了基础。
Magnaporthe oryzae is an aspartate mating ascomycete, but so far, there are relatively few studies on its sexual generation process and structure. In this study, two strains of Magnaporthe grisea, Guy-11 and 2539, were used in hybridization experiments on a variety of media to observe the sexual generation. The results showed that both strains could produce ascochlea after hybridization on all the test medium, but the number, production speed and maturity of the two strains were different, and the best was oat medium. In order to further observe the structure of the sexual generation, we observed the ascus and ascospores using Karl Fischer and Nile Red staining and fluorescence. Under the fluorescence microscope, the cell walls of ascus and ascospores were all clearly bright blue with Carbazone, and the cell structure was clear and distinct. Mature ascus shell can produce a large number of ascus, the ascus contains eight ascospores, ascospores usually contain 4 cells. At the same time, ascospores can be dyed with Nile red orange, indicating that the ascospores stored a large amount of fatty substances. This study provided an effective method for the fluorescent staining of the sexual generation structure of Magnaporthe grisea, which laid the foundation for further research.