【摘要】：我國(guó)喀斯特洞穴的資源相對(duì)豐富,然而國(guó)內(nèi)對(duì)洞穴微生物資源的研究還處于探索初步階段。本課題研究主要運(yùn)用純培養(yǎng)方法,對(duì)中國(guó)貴州興義和荔波兩個(gè)地區(qū)喀斯特洞穴放線(xiàn)菌資源的多樣性及其分布進(jìn)行研究,并對(duì)分離到的放線(xiàn)菌菌株進(jìn)行抗菌活性檢測(cè),以期從中挖掘新穎的活性產(chǎn)物。實(shí)驗(yàn)選取貴州地區(qū)喀斯特洞穴環(huán)境中不同類(lèi)型樣品為研究材料,通過(guò)利用多種分離培養(yǎng)基,共分離純化得到458株放線(xiàn)菌。根據(jù)放線(xiàn)菌形態(tài)特征初步歸納后,對(duì)分離到的458菌株進(jìn)行初步鑒定,結(jié)果發(fā)現(xiàn):分離菌株分布于放線(xiàn)菌門(mén)放線(xiàn)菌綱下11個(gè)目22個(gè)科44個(gè)屬,共193個(gè)種;通過(guò)16SrRNA基因序列比對(duì),相似性低于98.5%的潛在新種15個(gè)。小單孢菌屬(Micromonospora)、紅球菌屬(Rhodococcus)、微桿菌屬(Microbacterium)、鏈霉菌屬(Streptomyces)和類(lèi)諾卡氏菌屬(Nocardioides)為主要分布群體,而小月菌屬、動(dòng)球菌屬、厄氏菌屬、大理石雕菌屬等稀有類(lèi)群也有分布。對(duì)分離獲得的218株放線(xiàn)菌菌株進(jìn)行抗菌活性初篩。結(jié)果顯示,在這81株活性菌株中有16株對(duì)設(shè)定的10種指示菌都有抑菌活性,表現(xiàn)出很強(qiáng)的抑制作用。有46株至少對(duì)3種以上的指示菌有抑菌作用。在這些活性菌株中,表現(xiàn)出較強(qiáng)抗菌活性的菌株多數(shù)為鏈霉菌,小單孢菌,在其它類(lèi)群中也有分布。結(jié)果表明,喀斯特洞穴放線(xiàn)菌資源蘊(yùn)藏著巨大的活性天然產(chǎn)物開(kāi)發(fā)與應(yīng)用潛力。選取三株潛在新分類(lèi)單元YIMA1136T,YIMA1135T和SYSUK10001T進(jìn)行了形態(tài)學(xué)、生理生化檢測(cè)、化學(xué)組分分析、DNA同源性檢測(cè)和系統(tǒng)進(jìn)化分析的分類(lèi)研究,確定了3個(gè)新種的分類(lèi)地位,將其命名為Nocardioides cavernae,Nocardia cavernae和Lentzea cavernae。本研究通過(guò)純培養(yǎng)方法對(duì)貴州喀斯特洞穴放線(xiàn)菌的多樣性進(jìn)行初步掌握,為貴州地區(qū)喀斯特洞穴放線(xiàn)菌資源的研究和利用開(kāi)發(fā)奠下了理論基礎(chǔ)。此外,分離菌株的抗菌活性篩選結(jié)果揭示了喀斯特洞穴中蘊(yùn)藏著大量具有產(chǎn)活性天然產(chǎn)物的菌株,為篩選結(jié)構(gòu)新穎、活性?xún)?yōu)越的活性天然產(chǎn)物提供特殊的菌種資源。
[Abstract]:Karst caves in China are relatively rich in resources. However, the study of cave microbial resources in China is still in the initial stage. In this paper, the diversity and distribution of actinomycetes in karst caves in Xingyi and Libo areas of Guizhou Province were studied by pure culture method, and the antimicrobial activity of actinomycetes isolated from karst caves was tested. In order to excavate new active products from it. In this experiment, 458 strains of actinomycetes were isolated and purified from different kinds of samples in karst cave environment in Guizhou province. According to the morphological characteristics of actinomycetes, the strain 458 was preliminarily identified. The results showed that the isolates were distributed in 194 species, 22 families and 44 genera of 11 orders and 22 families under actinomycetes, and the 16SrRNA gene sequence was compared with each other. 15 potential new species with a similarity of less than 98.5%. Micromonospora, (Micromonospora), (Rhodococcus), Microbacillus, (Microbacterium), Streptomyces, (Streptomyces) and Nocardia, (Nocardioides) are the main distribution groups, while the rare groups, such as Motococcus, Acinetococcus, Ehrlla, Dali Stonework, are also distributed. The antibacterial activity of 218 actinomycetes strains was screened. The results showed that 16 of the 81 strains had antimicrobial activity against 10 kinds of indicator bacteria, which showed strong inhibitory effect. At least 46 strains had inhibitory effect on more than 3 kinds of indicator bacteria. Most of the strains with strong antibacterial activity were Streptomyces micromonosporium and distributed in other groups. The results showed that the actinomycetes in karst caves had great potential for exploitation and application of active natural products. Three potential new taxonomic units, YIMA1136T,YIMA1135T and SYSUK10001T, were selected for morphological, physiological and biochemical analysis, chemical component analysis, DNA homology detection and phylogenetic analysis. The taxonomic status of three new species was determined and named Nocardioides cavernae,Nocardia cavernae and Lentzea cavernae.. In this study, the diversity of actinomycetes in karst caves in Guizhou was preliminarily grasped by pure culture methods, which laid a theoretical foundation for the research and exploitation of actinomycetes in karst caves in Guizhou. In addition, the screening results of antimicrobial activity of isolated strains revealed that a large number of strains with active natural products were found in karst caves, which provided special resources for screening active natural products with novel structure and superior activity.
【學(xué)位授予單位】：昆明理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：Q93

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