Method
| Media Preparation Media was mixed into 2 liter Erlenmeyer flask according to ingredients below V Media was autoclaved V Flask contents were poured into Petri plates and allowed to cool |
Environmental Isolation White clover was obtained from grounds on the University of Maryland at College Park campus V Roots that contained nodules were sectioned into 1 cm length V Root sections were immersed in chlorine solution V Root sections immersed in ethanol V Root sections were covered with sterile water to rinse V Water rinse was repeated 3 times V Roots with nodules were ground using a pestle in sterile water V Results in Rhizobium containing solution |
| Inoculation Rhizobium containing solution was transferred to 4 Tryptone Yeast Extract agar (TYA) plates and 4 Yeast Mannitol agar (YMA) plates V The plates were incubated at room temperature (25◦C) for 4 days |
| Confirmation of Organism Identity Colonies were examined for morphological characteristics V Performed simple stain, Gram stain, and capsule stain V Performed motility stab V Prepared wet mount and observed using dark field microscopy V Oxidation/fermentation, mannitol utilization, and dextrose utilization tests were performed V From these tests it was confirmed that a pure culture of Rhizobium had been attained |