CONTENT

1. Introduction
2. Aim
3. Theory

4.Procedure

5.Observations

6.Conclusion

7.Reference
 INTRODUCTION
Rhizobium plural form rhizobia are prokaryotes whose
main function involves the conversion of stable
nitrogen gas in the atmosphere to a biologically useful
form. Nitrogenase is an enzyme complex that reduces
dinitrogen to ammonia.

Rhizobium is a biofertilizer. Biofertilizers are

substances that contain microorganisms which
when applied to the soil increase the nutrient
content and enhance the plant growth. Rhizobium,
present in the root nodules of the leguminous
plants, add nitrogen to the soil which is supplied to
the plants to enhance their growth.
Rhizobium is a bacterium found in soil that helps in
fixing nitrogen in leguminous plants. It attaches to the
roots of the leguminous plants and produce nodules.
These nodules fix atmospheric nitrogen and convert it
into ammonia that can be used by the plant for its
growth and development

A huge amount of energy is consumed during the

nitrogen fixation and the nitrogenase enzymes are
irreversibly inactivated by oxygen.
Acetylene reduction assay is used to measure the
nitrogenase activity.

Rhizobium infects the roots of leguminous plants. They

are usually found in the soil and produce nodules after
infecting the roots of the leguminous plants. As a result
nitrogen gas is fixed from the atmosphere. This
nitrogen is made available to the plants that help in
their growth and development. When the legume dies
there will be a breakdown of nodules. As a result,
Rhizobium is released back to the cell where it can
infect a new host.
 AIM
➢ To study the role of Rhizobium bacteria in
biofertilizer

Theory
Nitrogen-fixing bacteria, microorganisms capable of
transforming atmospheric nitrogen into fixed
nitrogen (inorganic compounds usable by plants). More
than 90 percent of all nitrogen fixation is effected by
these organisms, which thus play an important role in
the nitrogen cycle.
The symbiotic nitrogen-fixing bacteria invade the root
hairs of host plants, where they multiply and stimulate
formation of root nodules, enlargements of plant cells
and bacteria in intimate association. Within the nodules
the bacteria convert free nitrogen to ammonia, which the
host plant utilizes for its development. To ensure
sufficient nodule formation and optimum growth
of legumes (e.g., alfalfa, beans, clovers, peas, soybeans),
seeds are usually inoculated with
commercial cultures of appropriate Rhizobium species,
especially in soils poor or lacking in the required
bacterium.
 MATERIALS

1. Cryema medium
2. Beaker and petridish
3. Sterile water
4. Testube
5. Ethyl alcohol
6. Scalpel
7. Glassrods
8. Forceps
 PROCEDURES

• Collecting of healthy root nodules from leguminous

plants.
• washing nodules with sterile water.
• Sterlized nodules are crushed using sterile rod in a
test tube containing sterile water.
• And finally the suspension in inoculated into
petriplate containing yeast extract Mannion Congo
red agar by spread plate technique.
• Incubated for 2 - 3 days at 28-30°C
 OBSERVATION
Large gummy colonies of rhizobia will appear on
YEMA plates within 4-5 days.
 CONCLUSION

➢ Rhizobium, present in the root nodules of the

leguminous plants, add nitrogen to the soil which is
supplied to the plants to enhance their growth.
 REFERENCE

❖https://www.microscopemaster.com/rhizobium.html