螞蟻內(nèi)共生放線菌的鑒定及抗菌活性產(chǎn)物研究
發(fā)布時間：2018-08-09 20:03
【摘要】：近年來科學(xué)家們在探尋新型抗生素的篩選途徑時,發(fā)現(xiàn)昆蟲內(nèi)生菌能夠產(chǎn)生許多種類多樣的次生代謝產(chǎn)物,尤其那些能夠幫助宿主抵御病原菌入侵的放線菌,其中一些具有新穎的結(jié)構(gòu)并表現(xiàn)出較好的生物活性。由于這些化合物能夠存在于真核宿主體內(nèi),且不危害宿主,所以它們很有可能對人類也沒有毒副作用。因此這些昆蟲內(nèi)生放線菌具有很大產(chǎn)生人類藥用的抗生素潛力,所以近年來昆蟲內(nèi)共生放線菌受到越來越多科學(xué)家的關(guān)注,昆蟲內(nèi)生放線菌也將是持續(xù)地發(fā)現(xiàn)新抗生素的重要領(lǐng)域之一,這對于對抗日益增多的抗藥病原菌和新型疾病具有重要意義。本研究以實驗室篩選得到的30株螞蟻內(nèi)生菌為材料,通過培養(yǎng)特征觀察、抗菌活性測試和16S rRNA序列測序結(jié)果分析,發(fā)現(xiàn)兩株潛在的放線菌新種3H-GS17~T和2H-TWYE14~T和一株具有良好抗菌活性放線菌1H-GS2,對它們分別進行了系統(tǒng)的分類學(xué)鑒定和活性物質(zhì)的分離鑒定,結(jié)果如下:(1)16S rRNA分析結(jié)果顯示菌株3H-GS17~T是珊瑚放線菌屬的一株菌,與Actinocorallia glomerata JCM 9376T(98.13%)和Actinocorallia longicatena JCM 9377~T(97.64%)的16S rRNA序列相似性最高。化學(xué)分類學(xué)特征也與Actinocorallia的特征一致。而DNA DNA雜交結(jié)果和表型分析結(jié)果顯示菌株3H-GS17~T明顯不同于與它相近的菌株,進一步的實驗證明菌株3H-GS17~T和珊瑚放線菌屬的其他菌株都不相同。因此確定菌株3H-GS17~T是珊瑚放線菌屬的一個新種,命名為Actinocorallia lasicapitis,典型菌株是3H-GS17~T(=DSM 100595T=CGMCC4.7282T)。(2)16S rRNA分析結(jié)果顯示2H-TWYE14~T是鏈霉菌屬的一株菌,與Streptomyces niveus NRRL 2466T(98.84%)的16S rRNA序列相似性最高。gyr B基因分析結(jié)果同樣顯示2H-TWYE14~T屬于鏈霉菌屬。化學(xué)分類學(xué)特征也與鏈霉菌屬的其他菌株相一致。而2H-TWYE14~T與S.niveus NRRL 2466T之間的DNA DNA雜交結(jié)果和表型分析結(jié)果顯示2H-TWYE14~T明顯不同于S.niveus NRRL 2466T。因此確定菌株2H-TWYE14~T是鏈霉菌菌屬的一個新種,命名為Streptomyces camponoticapitis,典型菌株是2H-TWYE14~T(=DSM 100523T=CGMCC 4.7275T)。(3)活體生物活性檢測顯示菌株1H-GS2對部分測試真菌具有較好的生物活性,將菌株進行發(fā)酵,發(fā)酵產(chǎn)物同樣顯示良好的真菌抑制活性。對發(fā)酵產(chǎn)物中的化合物進行分離純化后,利用現(xiàn)代光譜學(xué)進行結(jié)構(gòu)分析鑒定,最終得到化合物白諾菌素(Albonoursin)。
[Abstract]:In recent years, while searching for new antibiotic screening pathways, scientists have found that insect endophytes can produce a variety of secondary metabolites, especially actinomycetes, which can help their hosts withstand the invasion of pathogens. Some of them have novel structures and exhibit good biological activity. Since these compounds are present in eukaryotic hosts and do not harm the host, they are likely to have no adverse effects on humans. Therefore, these insect endophytic actinomycetes have great antibiotic potential to produce human medicine, so in recent years, more and more scientists pay attention to insect endophytic actinomycetes. Insect endophytic actinomycetes will also be one of the important fields for the continuous discovery of new antibiotics, which is of great significance against the increasing number of drug-resistant pathogens and new diseases. In this study, 30 strains of ant endophytic bacteria screened in laboratory were used as materials, and the results of antibacterial activity test and 16s rRNA sequencing were analyzed through observation of culture characteristics. Two potential new actinomycetes, 3H-GS17~T and 2H-TWYE14~T, and one actinomycetes with good antibacterial activity, 1H-GS2, were found. The results were as follows: (1) the results of 16s rRNA analysis showed that the strain 3H-GS17~T was a strain of actinomycetes, and had the highest similarity with the 16s rRNA sequences of Actinocorallia glomerata JCM 9376T (98.13%) and Actinocorallia longicatena JCM 9377T (97.64%). The characteristics of chemotaxonomy are also consistent with those of Actinocorallia. The results of DNA DNA hybridization and phenotypic analysis showed that 3H-GS17~T was different from other strains of actinomycetes. Further experiments showed that 3H-GS17~T and other strains of actinomycetes were different. Therefore, 3H-GS17~T is a new species of actinomycetes, named Actinocorallia lasicapitis. the typical strain is 3H-GS17~T (DSM 100595T=CGMCC4.7282T). (2) 16s rRNA. The results show that 2H-TWYE14~T is a strain of Streptomyces. The analysis of 16s rRNA sequence with Streptomyces niveus NRRL 2466T (98.84%) also showed that 2H-TWYE14~T belongs to Streptomyces. The chemotaxonomic characteristics are also consistent with other strains of Streptomyces. The results of DNA DNA hybridization and phenotypic analysis between 2H-TWYE14~T and S.niveus NRRL 2466T showed that 2H-TWYE14~T was significantly different from S.niveus NRRL 2466T. Therefore, the strain 2H-TWYE14~T is a new species of Streptomyces, named Streptomyces camponticapitis. the typical strain is 2H-TWYE14~T (2H-TWYE14~T 100523T=CGMCC 4.7275T). (3), which shows that 1H-GS2 has good bioactivity to some tested fungi. The fermentation products also showed good fungal inhibitory activity. After the separation and purification of the compounds in the fermentation products, the structure analysis and identification of the compounds were carried out by modern spectroscopy. Finally, the compound (Albonoursin). Was obtained.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R915
本文編號：2175167
[Abstract]:In recent years, while searching for new antibiotic screening pathways, scientists have found that insect endophytes can produce a variety of secondary metabolites, especially actinomycetes, which can help their hosts withstand the invasion of pathogens. Some of them have novel structures and exhibit good biological activity. Since these compounds are present in eukaryotic hosts and do not harm the host, they are likely to have no adverse effects on humans. Therefore, these insect endophytic actinomycetes have great antibiotic potential to produce human medicine, so in recent years, more and more scientists pay attention to insect endophytic actinomycetes. Insect endophytic actinomycetes will also be one of the important fields for the continuous discovery of new antibiotics, which is of great significance against the increasing number of drug-resistant pathogens and new diseases. In this study, 30 strains of ant endophytic bacteria screened in laboratory were used as materials, and the results of antibacterial activity test and 16s rRNA sequencing were analyzed through observation of culture characteristics. Two potential new actinomycetes, 3H-GS17~T and 2H-TWYE14~T, and one actinomycetes with good antibacterial activity, 1H-GS2, were found. The results were as follows: (1) the results of 16s rRNA analysis showed that the strain 3H-GS17~T was a strain of actinomycetes, and had the highest similarity with the 16s rRNA sequences of Actinocorallia glomerata JCM 9376T (98.13%) and Actinocorallia longicatena JCM 9377T (97.64%). The characteristics of chemotaxonomy are also consistent with those of Actinocorallia. The results of DNA DNA hybridization and phenotypic analysis showed that 3H-GS17~T was different from other strains of actinomycetes. Further experiments showed that 3H-GS17~T and other strains of actinomycetes were different. Therefore, 3H-GS17~T is a new species of actinomycetes, named Actinocorallia lasicapitis. the typical strain is 3H-GS17~T (DSM 100595T=CGMCC4.7282T). (2) 16s rRNA. The results show that 2H-TWYE14~T is a strain of Streptomyces. The analysis of 16s rRNA sequence with Streptomyces niveus NRRL 2466T (98.84%) also showed that 2H-TWYE14~T belongs to Streptomyces. The chemotaxonomic characteristics are also consistent with other strains of Streptomyces. The results of DNA DNA hybridization and phenotypic analysis between 2H-TWYE14~T and S.niveus NRRL 2466T showed that 2H-TWYE14~T was significantly different from S.niveus NRRL 2466T. Therefore, the strain 2H-TWYE14~T is a new species of Streptomyces, named Streptomyces camponticapitis. the typical strain is 2H-TWYE14~T (2H-TWYE14~T 100523T=CGMCC 4.7275T). (3), which shows that 1H-GS2 has good bioactivity to some tested fungi. The fermentation products also showed good fungal inhibitory activity. After the separation and purification of the compounds in the fermentation products, the structure analysis and identification of the compounds were carried out by modern spectroscopy. Finally, the compound (Albonoursin). Was obtained.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R915
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，本文編號：2175167
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