發(fā)布時(shí)間：2018-05-31 22:12

  本文選題：胡楊林 + 放線菌　； 參考：《塔里木大學(xué)》2015年碩士論文

【摘要】：和田地區(qū)位于新疆維吾爾自治區(qū)最南端，其地理位置特殊，孕育了較為獨(dú)特的生態(tài)系統(tǒng)，，在此生態(tài)環(huán)境中存在著具有沙漠英雄美譽(yù)的原始胡楊林；胡楊林地處干旱、缺水、高溫、高輻射的環(huán)境中，其土壤中可能蘊(yùn)藏的較為特殊的微生物群落，因此本研究以新疆和田地區(qū)胡楊林土壤作為研究材料，對(duì)此環(huán)境中的放線菌資源進(jìn)行研究，以期發(fā)現(xiàn)較為原始和獨(dú)特的放線菌物種資源；同時(shí)在此資源中尋找有價(jià)值的放線菌菌種，對(duì)其代謝產(chǎn)物種類和功能進(jìn)行研究探索。 1.土壤放線菌分離鑒定及拮抗菌株篩選 以采自新疆和田地區(qū)胡楊林11個(gè)采集點(diǎn)的土壤混合樣品為研究材料，使用10種本實(shí)驗(yàn)室的經(jīng)典培養(yǎng)基分離獲得108株隸屬于3個(gè)屬38個(gè)種的放線菌。分別為鏈霉菌屬（Streptomyces）糖絲菌屬（Saccharothrix）和諾卡氏菌屬（Nocardiopsis），其中鏈霉菌屬為優(yōu)勢(shì)放線菌類群，涵蓋35個(gè)種。對(duì)分離鑒定的38個(gè)種的放線菌菌株以革蘭氏陽(yáng)性（金黃色葡萄球菌Staphylococcus aureus、表皮葡萄球菌Staphylococcus epidermidis）、革蘭氏陰性（大腸桿菌Escherichia coli）病原菌，人體病原真菌（白色念珠菌Candida albicans）、農(nóng)業(yè)病原真菌（棉花枯萎病原真菌Fusarium oxysporum f. sp. Vasinfectum）、棉花黃萎病原真菌(Verticillium dahliae)、紅棗黑斑病原真菌（Alternaria alternate Keissle）為靶標(biāo)菌進(jìn)行拮抗菌株篩選，最終獲得有18株菌株具有抑菌效果，其中1株菌株對(duì)金黃色葡萄球菌有抑制效果的，9株菌株對(duì)表皮葡萄球菌有抑制效果；4株菌株對(duì)白色念珠菌有抑制效果；2株菌株對(duì)紅棗黑斑病原真菌存在抑制作用；沒(méi)有獲得對(duì)大腸桿菌和棉花枯萎病、黃萎病原真菌有抑菌作用的菌株。 2.胡楊林土壤潛在新種TRM43006多相分類研究 通過(guò)形態(tài)特征、生理生化的特征、全細(xì)胞化學(xué)特征以及基因型特征等多相分類方法，對(duì)分離于和田地區(qū)胡楊林土壤放線菌TRM43006進(jìn)行分類鑒定，最終確定菌株TRM43006為鏈霉菌屬中的一個(gè)新種，將其命名為產(chǎn)吲哚鏈霉菌（Streptomyces indologenes）。 3.胡楊林放線菌TRM43006次生代謝產(chǎn)物分離鑒定 對(duì)具有金黃色葡萄球菌、表皮葡萄球菌抗菌活性的放線菌新物種TRM43006進(jìn)行了次生代謝產(chǎn)物研究。應(yīng)用大孔樹(shù)脂富集、薄層色譜、硅膠柱色譜、Sephadex LH-20凝膠柱色譜等方法對(duì)該菌株代謝產(chǎn)物進(jìn)行分離純化，獲得3個(gè)化合物。通過(guò)1D2D NMR波譜數(shù)據(jù)分析，鑒定化合物結(jié)構(gòu)分別為β-羰基-3-吲哚-丙醛（1）、鄰苯二甲酸二（2-乙基）已酯（2）、13-甲基肉豆蔻酸（3），其中13-甲基肉豆蔻酸對(duì)表皮葡萄球菌生物膜形成具有較強(qiáng)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.
【學(xué)位授予單位】：塔里木大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S714.3

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