| NaNO3 solution (25 g/L) | 30 ml |
| P-IV metal solution | 6 ml |
| CaCl2·2H2O solution (2.5 g/L) | 10 ml |
| MgSO4·7H2O solution (7.5 g/L) | 10 ml |
| K2HPO4 solution (7.5 g/L) | 10 ml |
| KH2PO4 solution (17.5 g/L) | 10 ml |
| Vitamin B12 solution | 1 ml |
| Soilwater: GR+ medium | 40 ml |
| Deionized water to | 1 L |
Vitamins are added after autoclaving.
P-IV metal solution:
| Na2EDTA·2H2O | 0.75 g |
| FeCl3·6H2O | 0.097 g |
| MnCl2·4H2O | 0.041 g |
| ZnCl2 | 0.005 g |
| CoCl2·6H2O | 0.002 g |
| Na2MoO4·2H2O | 0.004 g |
| Deionized water to | 1 L |
To approximately 950 ml of dH2O, add ingredients in order listed stirring continuously. The EDTA should be fully dissoled before adding other components. Bring to 1 L with dH2O and store at 4 C.
Vitamin B12 solution:
| HEPES buffer | 2.4 g |
| Vitamin B12 | 0.027 g |
| Deionized water to | 200 ml |
Prepare 200 ml of HEPES buffer at pH 7.8 and fully dissolve vitamin. Filter-sterilize and store in dark at -20 C.
Soilwater: GR+ Medium:
| Greenhouse soil | 1 tsp |
| CaCO3 | 1 mg |
| Deionized water to | 200 ml |
Combine ingredients. Cover container and steam for 2 consecutive days, 3 h each day with gradually rising temperature to 95 C in 15 min, then to 98 C for 3 h. Cool at room temperature and store at 4 C.
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