Isolation, Identification and Extracellular Enzymatic Activity of Culturable Extremely Halophilic Archaea and Bacteria of IncheBoroun Wetland

Abstract

Extremely halophilic diversity of IncheBroun wetland located in the north of Iran was investigated by using culture-dependent methods. Sampling was carried out in May and September 2014. In each sampling 4 distinct regions of wetland were analyzed by using complex media like MGM, JCM168, MH1 and an alkaliphilic medium containing 23% salts. After incubation at 40°C, a total of 406 isolates and 2.1 × 106CFU/ml were obtained in culture media. Among them 361 isolates were obtained from MGM and 39 isolates from JCM 168, 3 isolates from MH1 and 3 isolates from the alkaliphilic media. Initial morphological, biochemical and physiological tests were performed. Production of 4 hydrolytic enzymes by 45 selected strains was assayed qualitatively. A total of 38, 19 and 6 strains were able to produce lipase, DNase and amylase activity. Protease activity was not observed among strains. As total 45 strains were selected randomly and phylogenetic analysis of 16S rRNA was performed for them. Among selected strains 40 isolated strians belonged to Haloarchaea and were belonged to the genera:Haloarcula(30%),Halorubrum(27.5%),Haloferax(17.5%),Halobellus(10%),Halogeometricum(5.2%),Halobacterium(2.6%),Halolamina(2.6%),Halorhabdus(2.6%) andHalostagnicola(2.6%).HaloarculaandHalorubrumwere the dominant populations. A total of 5 strains belonged to domain ofBacteriaand were similar to members ofRhodovibrio(40%),Pseudomonas(40%) andSalicola(20%).