Name: Oculares, Arlimaze Breathe M.

Section Code: A181

Laboratory Activity No. 4

Rhizobium (Nitrogen-fixing bacteria)

Rhizobium is the most well-known species of a group of bacteria that acts as the primary symbiotic fixer
of nitrogen. These bacteria can infect the roots of leguminous plants, leading to the formation of lumps or
nodules where the nitrogen fixation takes place. The bacterium’s enzyme system supplies a constant source of
reduced nitrogen to the host plant and the plant provides nutrients and energy for the activities of the bacterium.

Objectives:
1. To identify the bacteria according to morphology.

Materials:
Small scissors/sharp knife
Root nodules
Gram-stain/Iodine

Procedure
1. Wash root nodules with water to remove all traces of soil.
2. Put a root nodule on a glass slide and cut open using a small scissors/sharp knife and observe the color.
What is the color of the nodule and what does it indicate? Pink/Red, It appears to be pink or red in color
because of the presence of a pigment called leghemoglobin. The presence of iron in this pigment accounts for its
pinkish color. Leghemoglobin promotes the function of the nitrogenase enzyme during nitrogen fixation by
scavenging the oxygen.
3. Crush the nodule using the slide and put a cover slip. Observe under the microscope.
What do you observe? I can see a very tiny rhizobium inside the crushed nodule.
4. Drop an iodine or gram-stain to the crushed nodule.
5. Take a picture and identify the bacteria.
RHIZOBIUM
(Nitrogen-fixing bacteria)