Isolation Method And Systematic Taxonomy Study On Streptomyces From Hanxing Iron Mine

Three kinds of known media, M5, M5', tap-water agar were used for streptomycetes isolation from typical magnetite samples collected from Hanxing Iron mine area as well as and iron-sap agar, which was designed by ourselves. The media were supplemented with 0.125% of lycine or sodium pyruvate to promot the growth respectively, and all of the media were supplemented with potassium dichromate as the inhibitor for bacteria.The result showed that for different kinds of samples, the kinds and amounts of actinobacteria colonies isolated on various media and supplements were quite different. Iron-mine-sap agar was the best isolation medium for all the samples, and adding lycine or sodium pyruvate can improve the streptomyces isolation rate efficiently, potassium dichromate can not inhibit the growing of bacteria.In this research, 16S rDNA phylogenetic method, morphology, physiological and biochemical features, cell chemical composition analysis and numerical classification were used for systematics of 43 typical strains and they were distributed into Streptomyces,sharing similarities with 19 nearest known neighbors.10 strains such as 163001 and 163005 ect have the identical sequence with the known type strain, the other 33 strains sharing 98.31%~99.92% similarities of 16S rDNA sequence with their nearest known neighbors. Strains 195004 and 195001 have the similarities of 99.47% and 99.39% with Streptomyces spinoverrucosus, respectively. Strain 034005 has the similarity of 99.09% with Streptomyces rubrogriseus . Strain 195005 has the similarity of 99.77% with Streptomyces parvus. Strain 195011 has the similarity of 99.92% with Streptomyces microflavus. Strain 035035 has the similarity of 98.07% with Streptomyces griseoloalbus. Strain 035005 has the similarity of 98.39% with Streptomyces fragilis, and strains 032004 and 032007 have the similarities of 99.98% and 99.70% with Streptomyces fradiae.Based on the above result, the culture features and morphological characters of the test strains on there culture media were observed by using coverslip techniques. In the physiology and biochemistry experiment , carbon utility, nitric acid deoxidiation,antibiotic sensitivity and NaCl tolerance were studied. The result of chemotaxonomy showed: all the strains contained L,L-DAP but does not contained meso-DAP in their cell walls. All the strains do not contain characteristic sugar (Sugar type C). Chemical composition of cell wall is type I. All the phenotype features are consistent with their phylogeny position of Streptomyces. Numerical classification analysis showed that all strains, except for strains 032004, 195008, 195009, could be divided into 13 groups by 91% similarities, compared with the result of phylogenetic analysis, 10 groups are the same, which showing a very high relavance.