Preparation of solidified medium M7
| - | Stock solution (%) | M7 (ml) 3 | | KNO3 | 1 | 10 | | MgSO47H2O | 0.1 | 10 | | (NH4)2 HPO4 | 0.2 | 5 | | CaSO4 | satured solution | 10 | | Soil extract 1 | - | 20 | | Sphagnum extract 2 | - | 10 | | Distilled water | - | 930 | | Micronutrient | - | 5 |
Add 1.5% agar before autoclaving. 1 Soil extract:
Weight 200 g of garden soil without great humus content, fertilizers or other protective agents. Add 1 liter of deionized water, and sterilize by heating for 1 hour.
Separate the decanted extract from particules by centrifugation. Fill into small containers of stock solution, autoclave for 1 hour at 1 atmosphere in three consecutive days. Store in the refrigerator.
2 Sphagnum extract:
Weight 50 g of dried Sphagnum, add 1.5 l of deionized water and sterilize by heating for 1 hour. Decant the extract. Fill into small containers and autoclave for 1 h at one atmosphere in three consecutive days. Store in the refrigerator. 3 Add vitamine B12 (5X10 -6 g) in sterile solution after autoclaving.
Composition of the micronutrient solution | - | Stock solution (%) | Micronutrient solution | | ZnSO4 7H2O | 0.1 | 1.0 ml | | MnSO4 4H2O | 0.1 | 2.0 ml | | H3Bo3 | 0.2 | 5.0 ml | | Co(NO3)2 6H2O | 0.02 | 5.0 ml | | Na2MoO4 2H2O | 0.02 | 5.0 ml | | CuSO4 5H2O | 0.0005 | 1.0 ml | | Distilled water | - | 981 ml | | FeSO4 7H2O | - | 0.7 g | | EDTA (Titriplex III, Merck) | - | 0.8 g |
Autoclave separately two micronutrient solutions: Sol. I - 881 ml distilled water + stock solutions of salts whithout FeSO4 + 0.4 g EDTA
Sol. II - 100 ml distilled water + 0.7 g FeSO4 + 0.4 g EDTA. Combine after cooling. |
|