Ramularia beticola Fautrey et Lambotte
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1. To thaw a frozen ampoule, place in a 2530 °C water bath, until just thawed (approximately 5 minutes). Immerse the ampoule just sufficient to cover the frozen material. Do not agitate the ampoule.
2. Immediately after thawing, wipe down ampoule with 70% ethanol and aseptically transfer 10 microliter (or any amount desired up to all) of the content onto a plate or broth with medium recommended.
3. Incubate the inoculum/strain at the temperature and conditions recommended.
4. Inspect for growth of the inoculum/strain regularly. The sign of viability is noticeable typically after 5-7 days of incubation. However, the time necessary for significant growth will vary from strain to strain.
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Hestbjerg H, et al. Development of Ramularia leaf spot on sugar beet as influenced by temperature and the age of the host plant. J. Phytopathol. 140: 293-300, 1994.