和田胡杨林土壤放线菌分离、鉴定及次生代谢产物研究

发布时间：2018-05-31 22:12

本文选题：胡杨林 + 放线菌　；参考：《塔里木大学》2015年硕士论文

【摘要】：和田地区位于新疆维吾尔自治区最南端，其地理位置特殊，孕育了较为独特的生态系统，，在此生态环境中存在着具有沙漠英雄美誉的原始胡杨林；胡杨林地处干旱、缺水、高温、高辐射的环境中，其土壤中可能蕴藏的较为特殊的微生物群落，因此本研究以新疆和田地区胡杨林土壤作为研究材料，对此环境中的放线菌资源进行研究，以期发现较为原始和独特的放线菌物种资源；同时在此资源中寻找有价值的放线菌菌种，对其代谢产物种类和功能进行研究探索。 1.土壤放线菌分离鉴定及拮抗菌株筛选以采自新疆和田地区胡杨林11个采集点的土壤混合样品为研究材料，使用10种本实验室的经典培养基分离获得108株隶属于3个属38个种的放线菌。分别为链霉菌属（Streptomyces）糖丝菌属（Saccharothrix）和诺卡氏菌属（Nocardiopsis），其中链霉菌属为优势放线菌类群，涵盖35个种。对分离鉴定的38个种的放线菌菌株以革兰氏阳性（金黄色葡萄球菌Staphylococcus aureus、表皮葡萄球菌Staphylococcus epidermidis）、革兰氏阴性（大肠杆菌Escherichia coli）病原菌，人体病原真菌（白色念珠菌Candida albicans）、农业病原真菌（棉花枯萎病原真菌Fusarium oxysporum f. sp. Vasinfectum）、棉花黄萎病原真菌(Verticillium dahliae)、红枣黑斑病原真菌（Alternaria alternate Keissle）为靶标菌进行拮抗菌株筛选，最终获得有18株菌株具有抑菌效果，其中1株菌株对金黄色葡萄球菌有抑制效果的，9株菌株对表皮葡萄球菌有抑制效果；4株菌株对白色念珠菌有抑制效果；2株菌株对红枣黑斑病原真菌存在抑制作用；没有获得对大肠杆菌和棉花枯萎病、黄萎病原真菌有抑菌作用的菌株。 2.胡杨林土壤潜在新种TRM43006多相分类研究通过形态特征、生理生化的特征、全细胞化学特征以及基因型特征等多相分类方法，对分离于和田地区胡杨林土壤放线菌TRM43006进行分类鉴定，最终确定菌株TRM43006为链霉菌属中的一个新种，将其命名为产吲哚链霉菌（Streptomyces indologenes）。 3.胡杨林放线菌TRM43006次生代谢产物分离鉴定对具有金黄色葡萄球菌、表皮葡萄球菌抗菌活性的放线菌新物种TRM43006进行了次生代谢产物研究。应用大孔树脂富集、薄层色谱、硅胶柱色谱、Sephadex LH-20凝胶柱色谱等方法对该菌株代谢产物进行分离纯化，获得3个化合物。通过1D2D NMR波谱数据分析，鉴定化合物结构分别为β-羰基-3-吲哚-丙醛（1）、邻苯二甲酸二（2-乙基）已酯（2）、13-甲基肉豆蔻酸（3），其中13-甲基肉豆蔻酸对表皮葡萄球菌生物膜形成具有较强抑制作用。
[Abstract]:The Hotan area is located at the southernmost end of the Xinjiang Uygur Autonomous Region, which has a special geographical position and is pregnant with a unique ecological system. In this ecological environment, the original poplar forest has a reputation for the hero of the desert. In the environment of drought, water shortage, high temperature and high radiation, the forest of Populus euphratica may contain more special microorganisms in the soil. In this study, the soil of Populus euphratica in Xinjiang and Hotan area was used as the research material to study the Actinomycetes Resources in this environment, in order to find out the more primitive and unique actinomycetes species resources, and to find valuable actinomycetes in this resource, and explore the species and function of its metabolites.
Isolation and identification of actinomycetes from 1. soil and screening of Antagonistic Strains
108 strains of actinomycetes belonging to 38 species of 3 genera and 38 species were isolated from the 11 sampling sites collected from the Populus euphratica forest in Xinjiang Hetian area. 108 strains belonging to 3 genera and 38 species were isolated. The genus Streptomyces (Saccharothrix) and Nocardia (Nocardiopsis) were isolated from the genus Streptomyces. The Streptomyces were isolated from the genus Streptomyces. The dominant actinomycetes group, covering 35 species. The 38 strains of actinomycetes for isolation and identification were Gram-positive (Staphylococcus aureus Staphylococcus aureus, Staphylococcus epidermidis Staphylococcus epidermidis), gram-negative (Escherichia coli) pathogens, and human pathogenic fungi (Candida albicans Candida albicans), and agriculture. The pathogenic fungi (Fusarium oxysporum f. sp. Vasinfectum of cotton wilt pathogen), cotton Verticillium wilt pathogenic fungi (Verticillium dahliae), red jujube black spot fungus (Alternaria alternate Keissle) as target bacteria were screened for antagonistic strains, and finally 18 strains had bacteriostasis effect, of which 1 strains were against golden yellow grape balls. The bacteria had inhibition effect, 9 strains had inhibitory effect on Staphylococcus epidermidis; 4 strains had inhibitory effect on Candida albicans; 2 strains had inhibitory effect on red jujube black spot pathogenic fungi; no Escherichia coli and cotton Fusarium wilt were obtained, and the pathogenic fungi of Verticillium wilt had bacteriostasis.
2. polyphasic taxonomy of potential new species of Populus euphratica forest soil TRM43006
Through the multi-phase classification method of morphological characteristics, physiological and biochemical characteristics, whole cell chemical characteristics and genotype characteristics, the TRM43006 of the soil actinomycetes isolated from the Hetian area of Populus euphratica was classified and identified. Finally, the strain TRM43006 was a new species of Streptomyces, which was named Streptomyces indologenes.
Isolation and identification of secondary metabolites from 3. TRM43006 of Populus euphratica
The secondary metabolites of TRM43006, a new actinomycete species with Staphylococcus aureus and Staphylococcus epidermidis, were studied. The metabolites of the strain were purified by macroporous resin enrichment, thin layer chromatography, silica gel column chromatography, and Sephadex LH-20 gel column chromatography, and 3 compounds were obtained by 1D2D NMR spectroscopy. Data analysis showed that the compounds were identified as beta carbonyl -3- indole propionic aldehyde (1), phthalic two formic acid two (2- ethyl) hexyl ester (2) and 13- methyl myristic acid (3), of which 13- methyl myristic acid had a strong inhibitory effect on the formation of Staphylococcus epidermidis biofilm.
【学位授予单位】：塔里木大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S714.3

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