【作者】 耿钰；

【导师】 张利平；

【作者基本信息】 河北大学 ， 微生物学， 2007， 硕士

【摘要】 本论文目的主要是研究硫铁矿中微生物分离方法,并通过对分离物的鉴定了解硫铁矿中微生物的类群分布。本文首先综述了硫铁矿中微生物研究的概况、研究进展、主要应用及当前微生物分类鉴定常用的技术方法,并对采样地的地理环境进行了简要介绍。本论文采用自行设计的不同培养基和分离方法,对采集自保定涞水县九龙镇硫铁矿不同样品中的微生物进行了分离。实验采用8种不同营养成分、不同pH值的培养基对110份样品进行分离,共分离得到放线菌45株,细菌12株,真菌16株。采用形态学、细胞壁化学组分分析和生理生化特性测定等方法,对45株放线菌进行了初步鉴定。并对其中5株菌的16S rDNA进行了全序列测定,系统发育分析表明,所测定的4株菌均为链霉菌属(Streptomyces);另一株与链霉菌属(Streptomyces)的Streptomyces indigoferus, Streptomyces herbaricolor和北里孢菌属(Kitasatospora)的Kitasatospora kifunensis16S rDNA序列相似性均为97.25%,其分类地位需要进一步的实验进行验证。本论文依据形态和生理生化指标,对所分离的细菌进行了初步鉴定,通过芽孢观察和接触酶实验结果表明,菌株46-1和菌株50-3具有芽孢,属于兼性厌氧杆菌；经形态和半固体琼脂穿刺法实验观察表明,菌株50-1、57-1、57-2、46-2为好氧性球菌,而菌株45-1、45-2、50-2、72-1、72-2为兼性厌氧性球菌；对嗜酸性细菌lsh-01进一步进行了分子分类鉴定,通过16S rDNA全序列测定,该菌株属于酸性硫杆菌属(Acidithiobacillus),与氧化亚铁酸性硫杆菌相似度最高,达98.91%。真菌的分离物的研究,参考《真菌鉴定手册》的鉴定标准,进行了形态观察和生理生化特性的测定。结果表明菌株F75-4是一株较为特殊的兼性自养嗜酸真菌,从形态上看属于酵母菌。更多还原

【Abstract】 This thesis mainly focuses on studying the methods of isolating microorganisms from pyrite and identifying the distribution pattern of species in pyrite through identification of isolates. Firstly, a brief summary of this field has been made, then the progress and application of the research have been introduced. Moreover, the main methods currently applied to the identification of microorganisms have been presented and geographical environment of samples collection has also been briefly introduced.In this thesis, microorganisms of samples collected in the pyrite in Jiulong town, Laishui county of Baoding were isolated, using different self-designed media and isolating methods. Eight media of different nutrition ingredients and pH for isolating 110 samples were adopted in this experiment. The results showed that 45 strains of actinomycetes,12 strains of bacteria and 16 strains of fungi were sucessfully isolatedMorphological, cytochemical, physiological and biochemical features were determined for 45 strains of actinomycetes and the almost complete sequences of 16S rDNA were determined for 5 of them.In these five strains, four strains belonged to Streptomyce by phylogenetic analysis and another strain had the same similarity,97.25%, with Streptomyces indigoferus, Streptomyces herbaricolor and Kitasatospora kifunensis, so further experiments are needed in order to confirm its taxonomic status.This thesis primarily identified the bacteria isolated according to morphological, physiological and biochemical characteristics. According to the results of schaeffer-fulton method and catalase experiment, strain 46-1 and strain 50-3 contained endospore and produced catalase; according to the results of morphological observation and stab cultivation, strains 50-1、57-1、57-2、46-2 belonged to aerobic coccus and strains 45-1、45-2、50-2、72-1、72-2 belonged to facultative anaerobic coccus; Strain lsh-01 of acidophilic bacterium is proved to be a member of Acidithiobacillus through further almost complete sequences of 16S rDNA. The similarity of Strain lsh-01 with thiobacillus ferrooxidans reaches as high as 98.91%.The research of the fungi isolates involves the determining of morphological, physiological and biochemical features and the Handbook to Identification of Fungi was referred. Strain F75-4, a special facultative autotrophic acidophilic fungus iaolated, belonged to Saccharomyces.更多还原