2.2. Isolation of Halophilic Archaea from the Rhizospheric and Non-rhizospheric Soils of Salsola stocksii and Atriplex amnicola
Modified growth medium (MGM) with NaCl 250 g/L, casamino acids 7.5 g/L, peptone 5 g/L, yeast extract 2 g/L, KCl 30 g/L, trisodium citrate 3 g/L, MgSO4 24 g/L, pH 7.2; was used for the isolation and purification of archaea from rhizospheric and non-rhizospheric soils ofSalsola stocksii and Atriplex amnicola [30].These soil samples were collected from Khewra Salt Mine, Pakistan. For isolation of individual archaeal strains, the soil sample was mixed thoroughly, sieved and then one gram of the representative soil sample was used for further analysis. All samples were serially diluted upto 10-10 and 100 μL of each dilution from 10-3 to 10-6 were spread on MGM plates for counting colony forming units (CFU) per gram of soil (dry weight). Plates were incubated at 37 °C until the appearance of archaeal colonies. Single, well-isolated colonies were selected, grown in MGM broth overnight, and stored in 33% glycerol at -80 °C for subsequent characterization.