The first set of experiments dealt with the development of a selected
number of RMO (diazotrophs) pure cultures. They were
initially inoculated into liquid CCM medium (100 ml in 250 ml
capacity flasks), supplemented with NH4Cl (0.5 g1) and yeast
extract (0.2 g1), then incubated in a rotary shaker (100 rpm)
at 30 C for 24 h. Serial dilutions prepared from the resulting
liquid batch cultures were surface-inoculated on agar plates
prepared from the ice plant juice (crude and further successive
dilutions) and CCM for comparison. Inoculated plates were