【作者】 申慧彦；

【导师】 刘朝贵；

【作者基本信息】 西南农业大学 ， 蔬菜学， 2004， 硕士

【摘要】 花脸蘑[Lepista sordida (Fr.)Sing]为伞菌目(Agaricales)，口蘑科(Tricholomatceae)，香蘑属(Lepista)的一种野生腐生菌。在我国，野生花脸蘑广泛的分布在云南、四川、新疆、西藏、贵州和福建等地区，但花脸蘑在野生状态下，数量少，产量低，十分珍贵。其色泽美丽，气味浓香，味道鲜美，营养价值和医疗价值极高，是珍稀优质的野生食用菌。目前对花脸蘑的研究甚少，仅对其野生资源进行了调查，初步研究了花脸蘑的生物学习性以及驯化栽培，但关于花脸蘑生物降解的研究以及大规模人工栽培的报道尚未发现。 本论文从应用基础出发，对花脸蘑形态特征和生活史初次进行了较为系统的研究以及对菌丝体生长的营养和环境条件做了详细的试验，较系统的了解花脸蘑的生物学特性；从研究不同培养基和不同生长阶段的花脸蘑胞外酶活性入手，了解花脸蘑对不同培养基质的降解能力和不同生长阶段对培养枓的需求。以期为进一步研究花脸蘑的生理生化奠定理论基础和开发驯化花脸蘑提供科学依据。本试验做了如下研究： 花脸蘑菌丝生物学特性的研究 试验1 花脸蘑形态特征和生活史的研究 从分类学角度来说所有食用菌的宏观特征都是对它们自身进行分类鉴定的重要依据之一，通过对花脸蘑子实体、孢子印、孢子和菌丝体的观察详尽了解花脸蘑的形态特征以确立花脸蘑在植物学的分类地位，并通过借鉴同科的其它食用菌生活史以模拟花脸蘑生活史。 试验2 花脸蘑菌丝生长特性的研究 本试验通过采用固体培养对花脸蘑母种培养基进行筛选，从供试的五种培养基中选出最适培养基为综合PDA培养基，其次为小麦培养基。采用固体平板法对花脸蘑菌丝生长各个因-一-一一一-.—甲，居迪契牡犁玛拿处泌一—--一子进行优化，结果表明:花脸蘑菌丝生一长有很宽的碳源谱，生一长最适碳源为蔗糖，其次为淀粉:最适氮源为有机氮，酵母膏优于蛋白陈，无机氮源中硝态氮和铰态氮之间也有显著差异;菌丝生长适宜温度为25℃、pH值为6.5、碳氮比为20:l一60:1。并根据单因子试验采用正交试验设计对碳源、氮源、PH值进行了正交试验，结果表明碳源为蔗糖、氮源为酵母、PH值为7的组合菌丝生长最好。液体摇瓶培养当碳源为蔗糖时生物量最高，蔗糖浓度以5%时生物量最高，菌丝球的人小与糖类物质的利用率无必然联系。正交试验表明以碳源为蔗糖、氮源为蛋自陈、pH值为7的组合生物量最高。花脸蘑菌丝胞外酶活性的研究试验3不同碳源处理对花脸蘑菌丝胞外酶活性的影响 采用液体摇瓶试验测得花脸蘑菌丝生长时可以产生多种胞外酶:淀粉酶、纤维素酶、半纤维素酶、漆酶、过氧化物酶、愈创木酚酶，它们的酶活性不但和菌丝生物量有联系，还与反应底物(不同碳源)有关;反应底物(蔗糖)一致时，随着蔗糖浓度升高，各种酶活性升高，当蔗糖浓度为2%时各种酶活不再增加，但保持在较高水平，花脸蘑胞外酶活性对碳源蔗糖的饱和浓度为2%。试验4不同培养基对花脸蘑栽培种胞外酶活性的影响 本试验对六种培养基上的花脸蘑八种胞外酶:梭甲基纤维素酶、微晶纤维素酶、半纤维素酶、淀粉酶、漆酶、多酚氧化酶、愈创木酚酶和过氧化物酶活性进行测试，结果表明酶活性的动态变化与培养基的成分有必然的联系。同一酶系中菌丝所处的营养环境对酶活性大小的影响造成差异，并不影响酶活性的变化趋势。不同培养基对各种酶活性大小影响程度没有一致性，这可能是六种培养基中各种酶作用的底物含量不同的原因。花脸蘑栽培种供试培养基中，以粪草培养基菌丝生长速度最快，生长势好，菌丝颜色为淡紫色。从这六种培养基菌丝生长看花脸蘑为好气性真菌，当碳源为淀粉、“卜纤维素、纤维素时较易利用，对木质素的分解利用能力较差。试验5花脸蘑栽培过程中酶活性的变化 本试验从栽培种定植起到出现菇蕾结束，每隔五大取样，测其梭甲基纤维素酶、一1’=纤维素酶、淀粉酶、漆酌、多酚氧化酶、愈创木酚酶和过氧化物酶活性。试验结果表明:在花脸蘑栽培过程中，淀粉酶在菌丝生长初期阶段活性较高，当菌丝生长进入生理活跃期时，纤维素和、}几纤维素酶活性升高，说明这两种酶在花脸蘑营养生长阶段对碳源的提供起重要作用。当出现菌丝扭结，花脸蘑即将进入生殖期，纤维素和平纤维酶活达到一个高峰期，与木质素降解有关的四种酶:漆酶、多酚氧化酶、过氧化物酶和愈创木酚酶的活性也快速升高。更多还原

【Abstract】 Lepista sordida, which belongs to Agaricales, Tricholomatceae and Lepista, is one kind of the wild saprophytic fungus. In our country, the wild Lepista sordida is extensively distributed in the areas, such as Yunnan, Sichuan, Xinjiang, Tibet, Guizhou and Fujian, etc., but the output is extremely low, so it is very precious. Lepista sordida has beautiful color and luster, its smell is thick fragrant, its taste is delicious, its nutritive value and medical value are extremely high, it is a kind of rare and high-quality wild edible fungus. There is very few study on Lepista sordida at present, only some surveys about the wild resource, the preliminary research on the biological habit and tame culture of Lepista sordida has been done, but the research on Lepista sordida biodegradation and report of extensive artificial culture have not been found yet.From the viewpoint of application foundation, this thesis carries on comparatively systematic research on the life history of Lepista sordida for the first time, makes detailed experiments upon nutrition and environmental conditions that mycelium grows, so as to understand the biological characteristic of Lepista sordida detailedly and systemically. Start from studying the influence of different substrate and different stages of growing on extracellular enzymes activity of Lepista sordida, understanding the demand for the biodegradation ability of different substrates and the demand for substrates of different stages of growing, so as to establish the theoretical foundation of physiological biochemistry for studying Lepista sordida, and offer scientific basis to develop and tame Lepista sordida. This thesis has done the following researches:Research on the biological characteristic of Lepista sordida myceliumExperiment 1 Research on the form characteristic and life history of Lepista sordidaFrom the viewpoint of taxonomy, all macroscopical characteristics of the edible fungus are one of the important bases to classify and determine themselves. The research idea is following, throughthe observation on fruit body, spore print, spore and mycelium of Lepista sordida, in order to understand the form characteristic of Lepista sordida exhaustivly, and establish the classification status of Lepista sordida in botany. Besides, by referring to the life history of other edible fungus, so as to simulate the life history of Lepista sordida.Experiment 2 Research on the hypha growing characteristic of Lepista sordidaThis experiment adopts solid-state substrates to select the mother substrates of Lepista sordida. The comprehensive PDA substrate is most appropriate substrates, and second is the wheat substrates. The solid-state substrates is adopted to optimize the different growth factor of Lepista sordida, the results are following. The hypha of Lepista sordida has very wide carbon sources to grow, and the cane sugar is the best carbon source for growth, secondly it is the starch. The most appropriate nitrogen source is the organic nitrogen, and yeast lotion is better than albumen peptone. Besides, during the inorganic nitrogen sources, there is a remarkable difference between the nitrate nitrogen and ammonium nitrogen too. The suitable temperature for hypha growing is 25centigrade degree, pH value is 6.5, and the ratio of carbon and nitrogen is from 20:1 to 60:1. The orthogonal experimental design method to carbon source, nitrogen source and pH value is adopted according to single factor. The results indicate that the best combination for hypha growing is the cane sugar as carbon source, yeast as nitrogen source, and pH value is 7. As for the submerged culture with shake flask, carbon source has the most amount of biomass when cultivation, and it has the most mount of biomass when cane sugar density is 5%. The size of hypha ball and the utilization ratio of the candy material do not have positive connection. The orthogonal experimental shows, the combination of the cane sugar as carbon source, albumen peptone as nitrogen source, and pH value be equal to 7 has the更多还原