【作者】 孟祥贤；

【导师】 张志光； 陈作红；

【作者基本信息】 湖南师范大学 ， 应用真菌学， 2001， 硕士

【摘要】 本论文利用菌种分离技术、紫外吸收光谱技术、高效液相色谱技术、氨基酸组成分析技术、定性分析与定量分析技术、电镜技术、原生质体技术和脉冲电泳核型分析技术对古尼虫草(Cordyceps gunnii)的生物学特性及生理活性物质进行了研究，主要包括以下内容：古尼虫草菌种分离及生长曲线的测定、古尼虫草的扫描电镜观察、古尼虫草原生质体制备、脉冲电泳核型分析和古尼虫草的虫草氨基酸、虫草多糖、核苷类物质含量研究，研究结果如下： 1、优化筛选出了古尼虫草菌种分离的最佳部位及方法。 2、首次测定了古尼拟青霉菌丝体的生长曲线，并根据其生长规律，将其分为四个典型的生长时期：生长延迟期、对数生长期、稳定生长期及衰老期。 3、研究发现：古尼虫草菌丝体中含有丰富的虫草多糖、氨基酸及核苷类物质，其含量明显高于冬虫夏草，其中虫草多糖含量为28.2mg／g干菌丝，氨基酸含量为28.62mg／100mg干菌丝，胞苷、尿苷、腺苷、脱氧腺苷含量分别为：620.3μg／g、3784.32μg／g、2588.17μg／g、420.10μg／g，脱氧鸟苷、胸苷、虫草素微量，所测定的7种核苷类物质其总量为7402.4μg／g，是冬虫夏草的含量(1150.1μg／g)的6倍多。在氨基酸组成分析中，首次测得了古尼虫草中含有组氨酸和脯氨酸等17种氨基酸。 4、本研究探索出了古尼虫草菌丝体培养和原生质体制备的最佳条件，即在改良沙氏培养液中25℃，120rpm培养5天的菌丝体，最适于制备古尼虫草原生质体，2％的溶壁酶，30℃，120rpm酶解2小时为制备原生质体的最佳条件。 5、首次利用脉冲电泳技术研究了虫草的电泳核型，这也是首次利用该技术对虫草属真菌的核型研究。原生质体包埋块最适宜浓度为4.3×10~8个／mL，古尼虫草染色体DNA在电泳过程中迁移形成6条带，最大的染色体DNA分子量约为2100Kb，最小的约为600Kb。由此可以确认古尼虫草有6条染色体。更多还原

【Abstract】 The biological characteristics and some active component of cordyceps gunnii was studied for the first time ,from growth survey and drawing of the spawn, the observation cordyceps gunnii and paecilomyces gunnii through scanning electron microscope , electrophoretic karyotype of cordyceps gunnii, to the contentation of active components ,such as cordyceps polysaccharide , amino acid , nucleoside in the spawn,tranma and worm of cordyceps gunnii .the result showed as below :Four typical growth periods in the spawn were studied : the period of growth hesitation, the rapid growth period ,the growth steady period,and the old period.There were plenty of cordyceps polysaccharide , amino acid , nucleoside in the spawn of cordyceps gunnii , their contentation in it was below : cordyceps polysaccharide,28.2mg/g ; amino acid ,28.62mg/100mg ; cytidine ,620.3 g/g; uridine , 3784.32 g/g; adenosine , 2588.17 g/g ; deoxyadenosine , 420.1 g/g . The total contentation of 7 kinds of nucleoside was 7402.4 u g/g .His and Pro were measured in amino acide of cordyceps gunnii for the first time.Electrophoretic karyotype of Cordyceps gunnii was studied for the first time .The spawn of C.gunnii was cultured with sabouraud culture medium,which was kept at 25 and vibrated at a speed of 120rpm for five days were best suitable for preparation of protoplasts .It was the optimum condition to prepare protoplasts that spawn was digested in 2% lywallzyme at 30 for 2 h with vibration at a speed of 120 rpm .The concentation of 4.3 108 /mL of protoplasts was the best suitable for electrophoresis . Cordyceps gunnii was separated into 6 bands ,which the biggest estimated to be 2100kb ,the smallest estimated to be 600kb.更多还原