论文标题:利用RAPD， RFLP和生理生化方法对白灵菇栽培品种的分类研究
Studies on Classification of Pleurotus Nebrodensis with RAPD, RFLP and Physiology and Biochemistry Technique
论文作者 郑姣
论文导师 马向东,论文学位 硕士,论文专业 微生物
论文单位 河南农业大学,点击次数 131,论文页数 45页File Size6524k
2005-06-01论文网 http://www.lw23.com/lunwen_484415127/ 白灵菇;同工酶;RAPD;PCR-RFLP
Pleurotus nebrodensis; Esterase isoenzyme technology; RAPD; PCR-RFPL
本研究选用我国白灵菇的13株主栽品种和1株糙皮侧耳作为参照,应用菌丝及子实体形态特征、同工酶技术,并配合随机引物扩增DNA(RAPD)和聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术,从形态、生理生化、DNA三个层次对这些菌株进行研究,按其特征划分为不同的类型。 白灵菇菌株在PDA平板上的菌落生长及形态,与遗传有一定的相关性,但受外界环境因素的影响较大。白灵菇子实体的形态,虽然在一定程度上受外界环境的影响,但主要与遗传特性相关,大部分菌种表现出典型的白灵菇子实体特征。在生理生化研究中,酯酶同工酶最适合白灵菇菌种的分类研究,而过氧化物酶同工酶和多酚氧化酶同工酶的结果较差。在对DNA的分类研究中,RAPD技术较适合白灵菇的分类试验,它灵敏且重复性好,优于PCR-RFLP技术。本试验初步建立起以检测白灵菇DNA分子水平和酯酶同工酶技术为主,配合其他生理生化特征、形态特征的系列菌种检测技术。所有试验基本上都得到了一致的分类结果,说明有多种分类方法都可以应用到白灵菇的分类研究中。我国白灵菇主栽品种间遗传物质相似程度很高,表明现有栽培品种缺乏丰富的基因型,品种比较单一,市场上白灵菇命名的混乱情况,不仅有同种异名的情况,还有异名同种的现象,亟待整顿和规范。
Thirteen strains of Pleurotus nebrodensis and a strain of Pleurotus ostreatus collected from china were studied with the RAPD , isoenzyme technology,mycelial and fructification morphology.Identified at three levels of morphology,physiology & biochemistry and molecular biology,all strains of Pleurotus nebrodensis can be classified as different type.The mycelial morphology of Pleurotus nebrodensis in PDA plate are influnenced more greatly by invironmental condition than by their gentics. However, the fructification morphology of Pleurotus nebrodensis are opposed to mycelial morphology. In physiology and biochemistry methods, esterase isoenzyme technology can better classify Pleurotus nebrodensis than peroxidase isozyme and PPO isozyme technology. In molecular research, RAPD technology has higher delicacy than PCR-RFLP technology. A series of technology for the identification of strains with the RAPD technology and esterase isoenzyme technology, complemented with other physiology and biochemistry methods and morphology characteration were established in this tests. All the studies in this paper have the same results. This show that there are many ways to classify Pleurotus nebrodensis. Those strains of Pleurotus nebrodensis in China had accordant genetic type in high degree. So the cultivated strains of Pleurotus nebrodensis in China are few, and the naming of them is jumbled. Some strains with different name are the same one, but the other ones with the same name are the different strains in the market.
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