Isolation and identification of culturable extremely halophilic archaea of Inche-Boroun wetland

Abstract

Haloarchaeal diversity of Inche-Boroun wetland in north of Iran in Golestan province was investigated by using culture-dependent methods. Sampling was carried out in May and September 2010. In each sampling, 4 distinct region of wetland were analyzed by using complex media like MGM, JCM168, MH1 and an alkaliphilic medium containing 23% salts. After incubation at 40ºC, a total of 406 isolates were prepared and 2.1×106 CFU/ml were obtained in culture media. Among all isolates, 361 isolates were obtained from MGM and 39 isolates from JCM 168, 3 isolates from MH1 and 3 isolate from alkaliphilic media. Initial morphological, biochemical and physiological tests were performed. According to the results, 45 isolates were selected and phylogenetic analysis of 16S rRNA was performed for them. Among selected strains, 40 isolates belonged to Halobacteriacaea and were related to Haloarcula, Halorubrum, Haloferax, Halobellus, Halogeometricum, Halobacterium, Halolamina, Halorhabdus and Halostagnicola (respectively 30, 27.5, 17.5, 10, 5.2, 2.6, 2.6, 2.6 and 2.6 percent of Haloarchaeal strains). A total of 5 strains belonged to the kingdom of Bacteria and were related to Rhodovibrio, Pseudomonas and Salicola (respectively 40, 40 and 20 percent of bacterial strains). According to our results and the limited numbers of haloarchaeal genera that having been discovered until now, it seemed that the culturable prokaryotic populations in this hypersaline environment was diverse.

Keywords

• Halophilic archaea
• Biodiversity
• 16S rRNA gene