Phenetic and Phylogenetic Analysis of Streptomyces spp. Isolated from Hydrocarbons Polluted Soil

Abstract

The research aims to highlight the role of polyphasic classification, which includes both phenotypic and genetic classification to determine and stabilize species characteristics within the genus Streptomyces, which is difficult to classify due to the large number of species and the large heterogeneity of phenotypic traits, in addition to its role in several areas, including Environments through resistance to many salts of heavy elements present in environments contaminated with hydrocarbons and their participation in the disposal of pollutants. In this study 6 isolates of Streptomyces obtained from 20 hydrocarbons contaminated and garden soil have been isolated and identified through phenotypic & genotypic characteristics using  nucleotide sequence of the 16s rDNA gene by universal primers 27f & 1392r following strains were  identified to  two strains that similar to Streptomyces flavogriseus ATCC 33331 , two strains belongs to Streptomyces albus J1074 & two strains belongs Streptomyces coelicolor A3(2). Depending on 48 of phenotypic characteristic by using numerical taxonomy , average linkage method and simple matching coefficient (Ssm) using statistical program SPSS. The strains grouped to three clusters in hierarchical tree, the two strains belongs to Streptomyces coelicolor grouped in cluster A at similarity of 99%, the two strains belongs to Streptomyces albus grouped in cluster B at similarity of 95% & the two strains that belongs to Streptomyces flavogriseus   accumulate in cluster C at similarity of 95%. The color of colony of all strain was grey to white ,the aerial mycelium of  Streptomyces coelicolor was branched spiral & rectalflex, while   Streptomyces albus have branched condense spiral aerial mycelium , Streptomyces flavogriseus produced unbranched & rectalflex  aerial mycelium. all of strains produced earthy odor. The all isolates showed tolerant to Nickel sulphate, Zinc sulphate, Lead acetate& nitrate, Silver acetate, Titanium dioxide, Cobalt nitrate & chloride. Also all isolates showed sensitivety to Mercuric sulphate & chloride. The results showed that all of strains were sensitive to Ciprofloxacin10 μg, Gentamicin10μg, Vancomycin 30 μg, Amikacin 10 μg, Imipenem 10 μg, Tobramycin 10 μg. In phylogenetic tree the species clustered in three group also as in phenetic tree in similarity percent between (96.7- 99.4%)  by Clustal W program and Maximum likelihood method, using Mega5.