Hey everyone, I have been trying to isolate Azospirillium from soil samples using this media recipe:
Ingredients Gms / Litre
Part A -
Malic acid 5.000
Dipotassium hydrogen phosphate 0.500
Ferrous sulphate 0.500
Manganese sulphate 0.010
Magnesium sulphate 0.200
Sodium chloride 0.100
Bromo thymol blue 0.002
Sodium molybdate 0.002
Calcium chloride 0.020
Agar 1.750
Part B - Potassium hydroxide 4.000
Final pH ( at 25°C) 6.8±0.2
I put the media is put into air tight tubes, innoculate, then the Azospirillium is meant to turn the solution blue as it grows. Then you can check morphology under a microscope. Ive added pictures of my tubes.
I am just not getting any growth. Even with an Azospirllium brasilense pos control. Im guessing there is a practical aspect I am missing, but cannot find enough information on it. Is the volume wrong? Am I meant to get rid of any air in the tube so it is completely anaerobic? Just not sure.