Rye medium A (modified with anitbiotics)
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Purpose
Ingredients
| Ingredient | 1 L | 500 ml |
|---|---|---|
| Part 1 | ||
| Rye broth | 1000 ml | 500 ml |
| Glucose | 10 g | 5 g |
| Agar | 15 g | 7.5 g |
| Part 2 | ||
| Natamycin (95%) | 10 mg | 5 mg |
| or | ||
| Pimaricin liquid (2.5%) | 0.4 ml | 0.2 ml |
| Ampicillin sodium salt | 250 mg | 125 mg |
| PCNB ((Pentachloronitrobenzene) (Terraclor 75%)* | 50 mg | 25 mg |
| Rifampicin (rifampin, rifamcyin) solution†† | 1 ml | 0.5 ml |
| Hymexazole (Tachigaren) ‡ | 50 mg | 25 mg |
Instructions
- Spread rye seed out on a piece of white paper and pick out ergot sclerotia and other contaminants.
- Weigh out 100 grams of cleaned seed and place in a sieve and rinse under running water, drain.
- Place the rinsed rye seed into 500 cc of distilled water, bring to a boil and turn off the heat. Allow to steep for 30 minutes.
- Strain steeped rye seed through several layers of cheese cloth into a 1 liter graduated cylinder; bring the volume up to one liter with distilled water.
- Autoclave and cool to 50 °C, then add the ingredients from part 2 (the powders can be suspended in a 10 ml sterile water blank):
- Pour plates, turn off hood light and cover with foil tent while cooling, then protect from light in storage.
- As formulated product (as opposed to active ingredient)
†† Rifampicin solution: dissolve 10.0 mg rifampicin in 1 ml DMSO.
‡ Hymexazole is added for PARP+H
Notes
- Rifampicin is soluble in DMSO. Rifampin is soluble in DMSO and methanol. Rifamycin SV is soluble in methanol and ethanol.
- Pimaricin is a broad-spectrum antifungal antibiotic that inhibits the true fungi but not most members of the Pythiaceae. Once in solution, it is inactivated by light so that culture plates must be stored and incubated in the dark. Growth of true fungi indicates the inactivation of pimaricin. Concentrations above 10 mg a.i./liter inhibit oospore germination of Pythium and Phytophthora.
References
- Erwin, D. C. and Ribeiro, O. K. 1996. Phytophthora diseases worldwide. APS Press, St. Paul MN. 562 pp.
Contributed by
From the Culture Media for Plant Pathogenic Fungi and Bacteria, University of Massachusetts: Contributed by Robert L. Wick
