【摘要】：作為微生物中重要類群之一的放線菌,它可以產生許多具有抗菌活性的次級代謝產物。這些天然的活性產物在醫(yī)療行業(yè)、化工行業(yè)、農業(yè)以及環(huán)境污染治理方面都有廣泛的應用。所以,從放線菌中發(fā)現新型抗生素成為了當今研究的熱點。為了避免從已有的放線菌中的重復分離化合物,本研究選取了不同于傳統的分離放線菌的分離源:植物和植物根際土壤,以期發(fā)現放線菌新種進而發(fā)現新型化合物。本研究所選取的分離源是萬壽菊(Tagetes erecta L.)植株和植株根際土壤。經過多次反復分離,從萬壽菊植株和植株根際土壤共分離得到了37株放線菌。并且對分離出來的37株放線菌分別對2種細菌和10中真菌進行了活性測試,結果顯示:有5株放線菌的發(fā)酵液浸提物對兩種細菌的抑制直徑在15mm以上;有9株菌對2種以上真菌表現出較好抑制的作用,其中有2株放線菌的菌落對3種真菌的抑菌直徑在20mm以上,只有1株放線菌對4種真菌的抑菌直徑在20mm以上。對從萬壽菊植株和根際分離出的37株放線菌的菌落形態(tài)、顯微鏡制片觀察及活性測試的結果分析后,對NEAU-TWSJ13和NEAU-YJC4進行16S r DNA的基因測序。通過網站的比對和構建系統發(fā)育分析,NEAU-TWSJ13屬于鏈孢囊菌屬(Streptosporangium),NEAUYJC4屬于小鏈孢菌屬(Catellatospora)。其中,與NEAU-TWSJ13最相似的菌株是Streptosporangium fragile DSM 43847T,16S r RNA相似性是98.6%;與NEAU-YJC4 16S r RNA最相似的菌株是Catellatospora bangladeshensis JCM 12949T(98.7%)。通過與相似菌株進行形態(tài)特征比較、化學分類特征比較和分子分類特征比較,結果顯示NEAU-TWSJ13和NEAU-YJC4在和其屬菌的基本特征一致的情況下,還可以區(qū)別于屬內的其他菌株。由此說明,NEAU-TWSJ13是鏈孢囊菌屬的新種,NEAU-YJC4是小鏈孢菌屬的新種,分別命名為Streptosporangium subfuscum和Catellatospora tagetis,并分別將它們送到中國、德國、日本菌種保藏中心進行保藏,保藏號分別為NEAU-TWSJ13(CGMCC 4.7146和DSM 46724)和NEAU-YJC4(CGMCC 4.7176和JCM 30053)。由此可見,把植物和植株根際土壤作為樣本進行分離提高了發(fā)現新種的可能性,同時也為分離出新型活性物質奠定了基礎。
[Abstract]:As an important group of microorganisms, actinomycetes can produce many secondary metabolites with antibacterial activity. These natural active products are widely used in medical industry, chemical industry, agriculture and environmental pollution control. Therefore, the discovery of new antibiotics from actinomycetes has become a hot spot. In order to avoid repeated isolation of compounds from existing actinomycetes, different from the traditional isolation of actinomycetes, plant and rhizosphere soil were selected in order to find new species of actinomycetes and then to find new compounds. The isolated source selected in this study is (Tagetes erecta L.). Plant and plant rhizosphere soil. After repeated isolation, 37 strains of actinomycetes were isolated from the rhizosphere soil and plants of marigold. The activity of 37 strains of actinomycetes was tested on 2 species of bacteria and 10 strains of fungi respectively. The results showed that 5 strains of actinomycetes had inhibitory diameter of more than 15mm on the two bacteria. 9 strains of actinomycetes showed better inhibition to more than 2 kinds of fungi. Among them, 2 strains of actinomycetes had inhibitory diameters over 20mm on 3 kinds of fungi, and only 1 strain of actinomycetes had inhibitory diameters above 20mm on 4 kinds of fungi. The colony morphology of 37 strains of actinomycetes isolated from marigold plants and rhizosphere, the results of microscopical observation and activity test were analyzed, and the 16s r DNA gene sequencing of NEAU-TWSJ13 and NEAU-YJC4 was carried out. Through the comparison and phylogenetic analysis of the website, NEAU-TWSJ13 belongs to the genus Alternaria, (Streptosporangium), NEAUYJC4 belongs to the genus Alternaria, and it belongs to (Catellatospora). The most similar strain to NEAU-TWSJ13 was Streptosporangium fragile DSM 43847 TX 16S r RNA, and the most similar strain to NEAU-YJC4 16S r RNA was Catellatospora bangladeshensis JCM 12949T (98.7%). The results showed that NEAU-TWSJ13 and NEAU-YJC4 could also be distinguished from other strains in the genus when they were consistent with the basic characteristics of their genus by comparing their morphological, chemical and molecular taxonomic characteristics with similar strains. Therefore, NEAU-TWSJ13 is a new species of the genus Alternaria, and NEAU-YJC4 is a new species of Alternaria, named Streptosporangium subfuscum and Catellatospora tagetis, respectively, and sends them to the preservation centers in China, Germany and Japan respectively. The storage numbers are NEAU-TWSJ13 (CGMCC 4.7146 and DSM 46724) and NEAU-YJC4 (CGMCC 4.7176 and JCM 30053) respectively. It can be seen that using the rhizosphere soil of plants and plants as samples improves the possibility of finding new species and lays a foundation for the separation of new active substances.
【學位授予單位】：東北農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S681.9;S182

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