中华真地鳖肠道微生物和脂肪体共生菌的研究

发布时间：2018-10-23 12:19

【摘要】：中华真地鳖(Eupolyphaga sinensisWalker)是我国传统的药用昆虫。昆虫肠道中存在着众多微生物。肠道微生物在宿主生命活动中发挥着重要作用。为了解中华真地鳖肠道微生物多样性情况,应用Illumina HiSeq测序技术检测了3只中华真地鳖肠道原核生物16S rDNAV4变异区序列,采用分离培养法研究了中华真地鳖肠道可培养好氧及兼性厌氧细菌的组成。经过测序,从三个样品中检测到30个菌门、187个菌科、201个菌属。拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)和厚壁菌门(Firmicutes)是样品中的优势菌门。紫单胞菌科(Porphyromonadaceae)、瘤胃菌科(Ruminococcaceae)、鞘脂单胞菌科(Sphingomonadaceae)、毛螺菌科(Lachnospiraceae)、黄单胞菌科(Xanthomonadaceae)、肠杆菌科(Enterobacteriaceae)、理研菌科(Rikenellaceae)、伯克氏菌科(Burkholderiaceae)、假单胞菌科(Pseudomonadaceae)、Chitinophagaceae科、Sphingobacteriaceae科、拟杆菌科(Bacteroidaceae)、芽胞杆菌科(Bacillaceae)和Methanomassiliicoccaceae科这11个科是样品中的优势菌科。Candidatus Azobacteroides属、Parabacteroides属、伯克氏菌属(Burkholderia)、瘤胃球菌属(Ruminococcus)、Kaistobacter属、假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)和vadinCA11属这8个属是优势菌属。利用分离培养法,从中华真地鳖肠道中获得7株好氧及兼性厌氧细菌。经鉴定,这7株菌分别属于沙雷氏菌属(Serratia)、棒状杆菌属(Corynebacterium)、Lysinibacillus属、芽孢杆菌属、Oceanobacillus属、假单胞菌属和红球菌属(Rhodococcus)。分离培养过程中得到的沙雷氏菌菌株能产红色素。该红色素被初步鉴定为灵菌红素,菌株产色素情况受温度影响,并且其发酵液对金黄色葡萄球菌(Staphylococcus aureus)、大肠杆菌(Escherichia coli)、伤寒沙门氏菌(Salmonella typhosa)、铜绿假单胞菌(Pseudomonas aeruginosa)和鸡白痢沙门氏菌(Salmonella Pullorum)均无抑制作用。蜚蠊目(Blattodea)昆虫脂肪体中普遍存在蟑螂杆状体属(Blattabacterium)共生菌。为了解中华真地鳖脂肪体中的共生菌情况,采用Illumina HiSeq测序技术检测了中华真地鳖脂肪体16S rDNAV4区扩增产物。证实了中华真地鳖脂肪体中存在蟑螂杆状体属共生菌,还检测到Alistipes属等178个丰度较低的其它菌属细菌。
[Abstract]:(Eupolyphaga sinensisWalker) is a traditional medicinal insect in China. There are many microbes in the gut of insects. Intestinal microbes play an important role in host life. In order to understand the diversity of intestinal microorganism in Trionyx sinensis, Illumina HiSeq sequencing technique was used to detect the 16s rDNAV4 mutation region of three intestinal prokaryotes of Trionyx sinensis. The composition of aerobic and facultative anaerobic bacteria in intestinal tract of Trionyx sinensis was studied by isolation and culture. After sequencing, 30 phylum, 187 families and 201 genera were detected from three samples. (Proteobacteria) and (Firmicutes) are the dominant phylum in the sample. (Porphyromonadaceae), (Ruminococcaceae), (Sphingomonadaceae), (Lachnospiraceae), (Xanthomonadaceae), (Enterobacteriaceae), (Rikenellaceae), (Burkholderiaceae), Pseudomonas, (Pseudomonadaceae), Chitinophagaceae, Sphingobacteriaceae, The 11 families of Bacillus (Bacteroidaceae), (Bacillaceae) and Methanomassiliicoccaceae are dominant genera of. Candidatus Azobacteroides, Parabacteroides, (Burkholderia), (Ruminococcus), Kaistobacter, (Pseudomonas), (Bacillus) and vadinCA11. Seven strains of aerobic and facultative anaerobic bacteria were isolated from the intestine of Trionyx sinensis. The seven strains were identified as belonging to the genus (Serratia), Corynebacterium, (Corynebacterium), Lysinibacillus, Bacillus, Oceanobacillus, Pseudomonas and (Rhodococcus)., respectively. In the process of isolation and culture, the strain of Shareh can produce red pigment. The red pigment was preliminarily identified as penicillin, and the pigment production of the strain was affected by temperature. Moreover, the fermentation broth had no inhibitory effect on Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), Salmonella typhimurium, (Salmonella typhosa), Pseudomonas aeruginosa (Pseudomonas aeruginosa) and Chicken Salmonella pullorum (Salmonella Pullorum). (Blattabacterium) symbiotic bacteria are commonly found in the fat body of cockroach roaches in (Blattodea). In order to understand the symbiotic bacteria in the fat body of Trionyx sinensis, the amplification products of 16s rDNAV4 region of the fat body were detected by Illumina HiSeq sequencing. It was confirmed that there were symbiotic bacteria of roach roaches in the fat body of Trionyx sinensis, and 178 other bacteria with low abundance, such as Alistipes genus, were detected.
【学位授予单位】：四川师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q96;Q93

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