【作者】 刘新锐；

【导师】 谢宝贵；

【作者基本信息】 福建农林大学 ， 微生物学， 2008， 硕士

【摘要】 Pleurotus属是一类分布广泛的且具有食、药用价值的木腐生真菌,近年来侧耳的生产发展很快,种类较多但分类较为混乱。因此建立一套侧耳属种的鉴定技术具有重要意义。本研究利用ITS序列及ITS-RFLP分析,对福建省食用菌种质资源保藏管理中心的Pleurotus属12种食用菌、559个菌株进行分子鉴别技术研究,清除了同种异名和同名异种的菌株,并建立了侧耳属生物学种的鉴别技术。具体结果如下:1、供试侧耳的ITS区(含ITS 1,5.8S rDNA和ITS 2)序列的长度范围为630-667bp,其中肺形侧耳为630bp,红平菇为667bp。根据ITS序列,Pleurotus属12种食用菌分为鲍鱼菇、金顶侧耳、红平菇、虎奶菇、肺形侧耳(凤尾菇和秀珍菇)、杏鲍菇、白灵菇(白灵菇和阿魏蘑)和糙皮侧耳(小平菇和平菇)8类,与报道的生物学种相符。高温平菇是一个无效种。2、使用了AluⅠ、DraⅠ、EcoRⅠ、HaeⅢ、MaeⅠ、Sau3AⅠ、SphⅠ七种酶,对Pleurotus属12种食用菌、585个菌株酶切分析,有效的清理了混乱菌株。ITS-RFLP分析结果和部分菌株出菇的子实体相吻合。建立了以ITS大小,AluⅠ、HaeⅢ、MaeⅠ酶切的侧耳属生物学种鉴定技术。3、利用建立的侧耳属生物学种鉴定技术,对7个可能是侧耳属菌株进行ITS大小,AluⅠ、HaeⅢ、MaeⅠ和DraⅠ酶切分析,得到5个是侧耳属的菌株,2个非侧耳属的菌株。经ITS序列分析,证实ITS-RFLP鉴定的准确性,表明ITS-RFLP可用于侧耳属种的鉴定。4、本研究采用ITS序列及ITS-RFLP技术,对侧耳属的12个种亲缘关系进行了研究,结果表明,鲍鱼菇、金顶侧耳、红平菇和虎奶菇等与其它侧耳的亲缘关系较远。鲍鱼菇和囊盖侧耳,金顶侧耳与白黄侧耳,红平菇和桃红侧耳,凤尾菇、秀珍菇和肺形侧耳,糙皮侧耳和佛罗里达,分别是同一个种的不同名称。ITS-RFLP有很好的稳定性和重复性,对侧耳属菌株种性的快速鉴定具有指导意义。更多还原

【Abstract】 The genus Pleurotus is a kind of wide spread wood saprobiotic fungus with edible and medical function,and its production is growing fast in recent years,while its taxonomy is confused.So,it is great significance to found a identified technology for Pleurotus species.In this study,we utilize the ITS sequences and ITS-RFLP analysis for studying the molecular identified technology among the 559 Pleurotus strains,which belong to 12 species collected by Fujian Edible Fungi Germplasm Resource Collection Center,and weed out the strains which had the different names in the same specie or the same name in the different species,also we found a molecular identified approach for Pleurotus species.The results are as follows.1.The length of ITS(including 5.8S rDNA) region ranged from 630-667bp.The length in pleurotus pulmonarius and Pleurotus djamor were 630bp and 667bp,respectively.Based on the ITS sequences analysis,12 Pleurotus species could be assigned to 8 kinds,including 5 single species(P.cystidiosus,P.citrinopileatus,P.djamor,P.tuber-regium,amd P.eryngii) and 3 complex groups(P.pulmonarius,P.nebrodensis and P.ostreatus complexes).The 8 kinds correspond with biological species reported.P.cystidiosus is meaningless in practical production.2.By combining the restriction fragment length polymorphism(ITS-RFLP) obtained from digestions with 7 enzymes AluⅠ,DraⅠ,EcoRⅠ,HaeⅢ,MaeⅠ,Sau3AⅠand SphⅠ,the 585 strains belonging to 12 Pleurotus species were efficient classified.The results from ITS-RFLP were corresponded with morphological characters.Based on the ITS size,AluⅠ-RFLP,HaeⅢ-RFLP and MaeⅠ-RFLP,we found a molecular identified technology for Pleurotus species.3.We investigated 7 probable Pleurotus strains in ITS size,AluⅠ-RFLP,HaeⅢ-RFLP, MaeⅠ-RFLP and DraⅠ-RFLP based on the molecular identified technology for Pleurotus species.7 strains were efficient identified.It is conferred that ITS-RFLP was veracity for identifying Pleurotus species by comparing the ITS sequencing.The result shows ITS-RFLP can be applied in identifying Pleurotus species.4.With the result of ITS sequence and ITS-RFLP patterns,the lineage relationship among 12 Pleurotus species are analyzed.The result showed that P.cystidiosus,P.citrinopileatus,P. djamor,P.tuber-regium was more distant from other Pleurotus species.P.abalonus and P. cyslidiosus are the same species,as well as P.citrinopileatus and P.cornucopiae,P.djamor and P. salmoneostramineus,P.pulmonarius,P.sajor-caju and P.geesteranus,P.floridanus and P. ostreatus.Furthermore,ITS-RFLP has good stability and repeatability,and will be guidable significance for fast identification of Pleurotus species.更多还原