双孢蘑菇疣孢霉病的发病过程及病原菌的核相研究

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【目的】确定有害疣孢霉的传播途径,明确双孢蘑菇受有害疣孢霉侵染后发病症状和微观形态变化,以及有害疣孢霉的核相。【方法】将有害疣孢霉喷施于培养料及覆土材料的不同深度,观察记录双孢蘑菇的发病情况;将有害疣孢霉接种于不同生长阶段的双孢蘑菇子实体,观察记录其发病情况;使用光学显微镜及扫描电镜观察双孢蘑菇子实体受有害疣孢霉侵染前后的形态变化;通过DAPI(4′,6-二脒基-2-苯基吲哚)染色的方法对有害疣孢霉核相进行观察。【结果】将有害疣孢霉接种于培养料及覆土层的不同深度得到双孢蘑菇发病率如下:覆土层表面>覆土层中间>覆土与培养料交界处>培养料中间层;有害疣孢霉可以侵染双孢蘑菇的任意阶段,将其接种于原基直径小于3 mm子实体表面时,得到不能正常分化的“马勃状”组织;对有害疣孢霉的侵染过程进行观察得到:其孢子可粘附于双孢蘑菇表面,并萌发长出芽管,接种处双孢蘑菇表面产生褐色病斑,双孢蘑菇菌丝体发生质壁分离,最后菌丝体膨大,细胞壁变薄甚至溢裂,菌丝体内部中空;有害疣孢霉产生两种类型的分生孢子,Ⅰ类无隔膜含1个细胞核;Ⅱ类具1隔膜含2个细胞核,2个细胞核被隔膜分开;细胞核的第1次有丝分裂发生于分生孢子母细胞中;厚垣孢子由上下2个细胞构成,上胞中含有2个细胞核。下胞含1–2个细胞核。有害疣孢霉的厚垣孢子萌发可产生1–2个芽管,芽管中细胞核的数目不断变化,一般0–2个细胞核。【结论】双孢蘑菇受其侵染后发生显著的细胞学变化;我们对有害疣孢霉做遗传分析时,进行单孢分离需挑取无隔膜的分生孢子为实验材料进行遗传分析。 【Objective】 The objective of this study was to determine the pathogen of harmful M. sphaericus, to clarify the onset and microscopic morphological changes of A. bisporus after it had been infected by P. nidulans. 【Method】 The harmful Worm spores were sprayed on different depths of culture materials and soil-covering materials, and the incidence of Agaricus bisporus was observed and recorded. The harmful Mucor sporis was inoculated into the fruiting bodies of Agaricus bisporus at different growth stages and observed the incidence ; Morphological changes of Agaricus bisporus fruiting bodies before and after the infection by A. wilfordii were observed with optical microscope and scanning electron microscopy; and the morphological changes of harmful warts were detected by DAPI (4 ’, 6-diamidino-2-phenylindole) staining Spore mold phase was observed. 【Result】 The results showed that the incidence of Agaricus bisporus was as follows: the surface of soil cover> the middle of soil cover> the interface of soil cover and culture material> the middle layer of culture material; Inoculation of Agaricus bisporus at any stage, when inoculated on the surface diameter of less than 3 mm diameter of the original entity, can not be normal differentiation “Bobo ” organization; : The spores can be attached to the surface of Agaricus bisporus and germinate germinate tube, inoculation at the surface of Agaricus bisporus brown spot, Agaricus bisporus mycelium plasmolysis, and finally mycelium dilated, thinning of the cell wall or even Mycotoxins produce two types of conidia, Class I without a membrane containing a nucleus; Class II with 1 membrane containing 2 nuclei, 2 nuclei separated by the diaphragm; nucleus The first mitosis occurred in the conidia of the mother cells; Chlamydospores by two upper and lower cells, the upper cell contains two nuclei. The next cell contains 1-2 nuclei. Chlamydomonas vaginalis harmful spore germination can produce 1-2 germ tube, germ tube in the number of nuclear changes, usually 0-2 nuclei. 【Conclusion】 Agaricus bisporus significantly affected the cytology after it was infected. When we did the genetic analysis of the harmful M. sphaeriforme, single-spore isolation was carried out to select the non-membrane-derived conidia as the experimental material for genetic analysis.
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