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【目的】从白穗软珊瑚中分离和鉴定共附生放线菌,运用PCR技术对所分离的放线菌进行I型聚酮合酶(PKS)筛选,研究其次级代谢产物。【方法】使用11种培养基对白穗软珊瑚共附生放线菌进行分离、鉴定,构建16S rRNA基因系统发育进化树,以基于I型PKS的KS基因设计的简并引物对所分离放线菌进行基因筛选,对阳性菌株用3种培养基发酵检测,对目标菌株进行放大规模发酵分离鉴定次级代谢产物。【结果】从白穗软珊瑚中分离到20株共附生放线菌,包括链霉菌属10株、迪茨氏菌属2株和盐水孢菌属8株,筛选获得18株I型PKS阳性菌株,并从菌株Salinospora arenicola SH04中分离到化合物rifamycin S和rifamycin W。【结论】首次从珊瑚共附生环境中分离得到海洋专属性稀有放线菌盐水孢菌属,并以I型PKS基因筛选为指导,分离鉴定了聚酮类化合物rifamycins,为研究软珊瑚共附生可培养放线菌的多样性和基于基因筛选指导分离次级代谢产物提供了可靠依据。
【Objective】 The objective of this study was to isolate and identify actinomycetes from soft white coral and screen for the type I polyketide synthase (PKS) by PCR. 【Method】 The 16S rRNA gene phylogenetic tree was isolated and identified by using 11 kinds of media. The degenerate primers based on the KS gene of type I PKS were separated and identified The strains were screened for genes. The positive strains were fermented in three kinds of media and the secondary metabolites were isolated and identified by amplification and scale fermentation of the target strains. 【Result】 Twenty strains of actinomycetes were isolated from soft white coral, including 10 strains of Streptomyces, 2 strains of Dietrichia and 8 strains of Salina, and 18 strains of PKS were positive The strains rifamycin S and rifamycin W were isolated from the strain Salinospora arenicola SH04. 【Conclusion】 For the first time, marine endemic Rare Actinomycetes Salina species were isolated from co-epiphytic corals, and the polyketide rifamycins were isolated and identified under the guidance of type I PKS gene screening. In order to study the co-attachment of soft corals The diversity of viable culturable actinomycetes and the basis of genetic screening guide the separation of secondary metabolites provide a reliable basis.