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放线菌是微生物中重要的药用菌,其产生的抗生素是20世纪医学皇冠上最耀眼的明珠,但随着抗生素滥用,抗生素耐药现象日趋严重,因此从特殊生态环境中发现新放线菌物种和新抗生素克服耐药是科研人员正在尝试的途径之一,文章初步探究双河洞系统中阴河洞和杉林洞两个支洞的放线菌多样性,并对分离菌株的抗菌活性进行筛选。以纯培养方法获得放线菌,以16SrRNA基因序列比对,分析菌株类别,获得的菌株进行液体发酵,纸片琼脂扩散法测定其发酵产物对真菌和细菌的抗菌活性,同时用PCR技术进行NRPS、PKS I和PKS II抗生素生物合成基因的探测。从两个支洞样品中分离到的放线菌分布于7个科10个属,两个洞共66株放线菌中有32株的发酵产物在至少一个抗菌活性筛选中显示阳性,总阳性率为48.5%。抗生素生物合成基因的初步探测结果显示,31株菌存在至少一个生物合成基因,NRPS 28株,PKS I17株,PKS II 21株,其中12株同时具有3种生物合成基因。初步显示了双河洞系统蕴含丰富的放线菌资源,值得深入研究。
Actinomyces is an important medicinal bacterium in microorganisms. The antibiotics it produces are the most dazzling pearl in the medical crown of the 20th century. However, with the abuse of antibiotics, the phenomenon of antibiotic resistance becomes more and more serious. Therefore, the new actinomycetes found in the special ecological environment To overcome the drug resistance of bacteria and new antibiotics is one of the ways that researchers are trying to explore. The article initially explores the diversity of actinomycetes in the two branch holes of Yinhe Cave and Shanlin Cave in Shuanghe Cave System. The antibacterial activity of the isolated strains Screening. The actinomycetes were obtained by pure culture method. The strains were analyzed by 16S rRNA gene sequence. The obtained strains were subjected to liquid fermentation. The antibacterial activity of the fermented products against fungi and bacteria was determined by agar diffusion method. Meanwhile, NRPS, Detection of PKS I and PKS II antibiotic biosynthetic genes. The actinomycetes isolated from the two branch holes were distributed in 10 genera of 7 families. The fermentation products of 32 of 66 actinomycetes in two holes showed positive in at least one antibacterial activity screening, and the total positive The rate is 48.5%. The preliminary detection of antibiotic biosynthesis genes showed that there were at least one biosynthetic gene, 28 strains of NRPS, 21 strains of PKS and 21 strains of PKS II, of which 31 strains had three biosynthetic genes. It shows that Shuanghe cave system contains abundant actinomycetes resources, which deserves further study.