【作者】 黄红英；

【导师】 陈作红； 张志光；

【作者基本信息】 湖南师范大学 ， 植物学， 2004， 硕士

【摘要】 本文以发现于湖南长沙的我国新记录种——沼生花褶伞为实验材料,利用菌种分离技术、光镜技术、电镜技术、高效液相色谱技术、毒理实验技术、电生理等技术手段,对沼生花褶伞的生物学特性和毒性进行了研究。主要包括内容:沼生花褶伞的生态学和组织学特征,菌种分离培养,菌丝体及子实体荧光染色方法的建立,子实体的栽培,对植物和动物的毒性研究,毒素的分离和鉴定,获得了如下的结果: 1.通过野外生态调查和利用光学显微镜,电子显微镜对沼生花褶伞进行了生态学和组织学的研究。沼生花褶伞是一种群生的草腐生菌,在相对稳定的生态条件下,子实体发生适宜温度26～32℃,相对湿度50～80%,降雨量30%以上,温度是主导因子。每年在春末夏初高温阵雨之后为盛生长期。担孢子为柠檬状,表面光滑,黑色,菌褶菌丝有锁状联合,褶缘囊状体发达。 2.以碳源、氮源、温度、PH值单因子实验设计培养条件,对沼生花褶伞菌丝体进行培养。通过对菌丝体直径测定及生长势分析,优化筛选出沼生花褶伞菌种培养的最适培养基为牛粪汁培养基:蔗糖20g,蛋白胨10g,K₂HPO₄1g,KH₂PO₄0.5g,MgSO₄.7H₂O 0.5g,琼脂20g,加发酵牛粪浸出液至1000ml,最适温度为28℃,最佳PH值为7.8,并证明蛋白胨,尿素为最佳氮源,蔗糖和玉米粉为最佳碳源。 3.利用核荧光染料Heochst33258和隔膜荧光染料Clacolour white对沼生花褶伞菌丝体及子实体进行复合染色。Calcour white在沼生花褶伞菌丝体观察过程中的最佳浓度0.0005%,而核荧光染料Hoechst33258的最佳PH为7.0,最佳浓度为10μg/ml。爬片2/5菌丝体的核荧光染色为最佳。 4.通过组织分离,无菌操作切取菌柄与菌盖交接处的小块组织,更多还原

【Abstract】 The biological characteristics and toxicity of Panaeolus paludosus ,a new recorded Changsha Hunan species in China , were studied by means of tissue isolation, light microscopy, scanning electronic microscopy, reversed-phase high performance liquid chromatography, electro-physiological methods. The study included the ecological and organization characteristics of Panaeolus paludosus, fluorescent dying for mycelium, culturing frutification, the toxicity to plants and animals, isolating and identifying of toxin, and the results were as following:1. The ecological and organization characteristics of Panaeolus paludosus were studied by means of ecological observation in field and observation through light microscopy and scanning microscopy techniques. Panaeolus paludosus is a coprophilous fungi, which emerges and develops in relatively steady environment where average temperature, humidity and soil water content were 26~32℃, 50~80% and 30% respectively. Among the environmental factors, temperature is major factor. Panaeolus paludosus grows fast after shower in the late summer and early autumn when it is hot. Basidiospore has black color, lemon shape, and smooth surface. Clamp connection of the lamellae mycelium is often found. Cheilocystidia are abundant in gill.2. Mycelium of Panaeolus paludosus were cultivated based on single factor design carbon source, nitrogen source, tempreture, and pH.The optimal culture condition of Panaeolus paludosus was investigated by testing and analyzing mycelium diameter, and the best medium was compost medium. The optimun culture conditions were pH 7.8, culture temperature 28 °C. The compost medium was 1000ml compost juice including sucrose 20g, peptone lOg, K^HPCMg ? KH2PO40.5g , MgSO4.7H2O 0.5g. The best nitrogen compound was peptone, and carbamide, the best carbon compound was sucrose and corn flour.3. Mycelium and fruiting body of Panaeolus paludosus were repeatedly dyed by nuclear fluorescent dye Heochst33258 and dissepiment stain calcolour white. The best concentration of calcolour white in the study of mycelium was 0.0005%. The optimal pH of nuclear fluorescent dye Heochst33258 was 7.0, the best concentration was lOug/ml. Two fifths of mycelial growth on the cover slid showed the best effect of nuclear fluorescent dye.4. The strains were got by means of the isolation of tissue and the aseptic incision of the small tissue between the stipe and pileus.The compost medium suited for the growing of strains, the better medium of protospecies was wheat substrate, when cultured Rpaludosus, the dung and straw were artificial manure. Layer sow was used germination of P. paludosus. Fruiting body of Panaeolus paludosus were successfully cultivated in indoor environment. The optimal temperature for the development of fruiting body was lower than the one for the growth of mycelium, the water content of material for culture was about 60%, and the formation of fruiting body needed abundant light.5. The effect of different concentration of crude venom on the hypocotyl growth of Phaseolus aureaus’s seed were measured. The changing curve of the content of paddy chlorophyll and MDA demonstrates the concentration of crude venom more than 50mg/L can obviously restrain hypocotyl growth. Treated with 50mg/L extractive liquid of mycelium for ten days, the content of chlorophyll decreased by更多还原