Assignment Topic:

Nitrogen Fixation through different process and root nodules

formation.
Subject:
Biological Nitrogen Fixation.
Submitted to:
Dr. Adeel Anwar
Submitted By:
Sheikh Muhammad Ijaz
Reg. No. :
19-Arid-4235
Department:
Agronomy
Semester:
7th
PMAS ARID AGRICULTURE UNIVERSITY
RAWALPINDI.
What is Nodule:
The root nodule is a selective marker between legumes and leguminous
bacteria. Nodule formation is regulated by chemical signals between plants and microbes, and
is one of the most well-studied chemical communications.

Nitrogen in Air:
All living things need nitrogen for the synthesis of proteins, nucleic acids. Acids and other
nitrogen-containing compounds. Contains the Earth's atmosphere. About 80 percent nitrogen
gas. It cannot be used in this form by most organisms unless it is fixed, that is, reduced (with
hydrogen), to form ammonia. Green plants, the main producers of organic matter, use this
supply of fixed nitrogen to make proteins that enter and pass through the food chain.
Microorganisms (decomposers) break down proteins into excreta and dead. Organisms release
ammonium ions. These two processes form part of Nitrogen cycle.

Nitrogen Fixation process:

Nitrogen can be fixed in three ways

 Atmospheric fixation – The fixation occurs spontaneously due to lightning a small

amount only is fixed this way.
 Industrial fixation – ln this way Haber process, which is very energy inefficient, is used
to make nitrogen fertilizers.
 Biological fixation – In this process nitrogen-fixing bacteria that fix 60% of nitrogen gas.

Biological Nitrogen Fixation:

The reduction of nitrogen gas to ammonia requires energy. It requires 16 molecules of ATP and
a complex set of enzymes to break the nitrogen bonds so that it can combine with the
hydrogen. Its reduction can be written as:
N2 +3H ================= 2NH3
Rhizobium :
Rhizobium is the best-known species of a group of bacteria that act as primary symbiotic
nitrogen fixers.
These bacteria can infect the roots of leguminous plants, resulting in the formation of lumps or
nodules where nitrogen is fixed. The enzyme system of the bacterium provides the host plant
with a constant source of low nitrogen and the plant provides nutrients and energy for the
bacterium's activities. About 90% of beans can be nodulated.

Root nodules formation:

Gene sets in bacteria control different aspects of the nodulation process. A strain of Rhizobium
can infect some species of legumes but not others, eg the pea is the host plant of Rhizobium
leguminosarum biovarviciae. While clover acts as a host for R. leguminosarum biovar trifolii.
Specific genes determine which Rhizobium strain infects which legume. Even if a strain is able
to infect a pod, the nodules that form may not be able to fix nitrogen. Such rhizobia are called
non-functional. Effective stresses induce nitrogen-fixing nodules. Effectiveness is controlled by
a different set of genes from specific genes in bacteria. Nod genes direct different stages of
nodulation.
The initial interaction between the host plant and free-living rhizobia is the release of a variety
of chemicals into the soil by the root cells. Some of them encourage the growth of bacterial
populations in the area around the roots (rhizosphere). Reactions between the bacterial cell
wall and certain compounds in the root surface are responsible for rhizobia recognizing their
correct host plant and attaching to root hairs. Flavonoids secreted by root cells activate nod
genes in bacteria which then lead to nodule formation. The entire process of nodulation is
regulated by highly complex chemical communication between the plant and the bacteria.
Once attached to the root hairs, the bacteria secrete Nod factors. These stimulate the hair to
curl. The rhizobia then invade the roots through the tips of the hairs where they form an
infection thread. This thread is produced by root cells, not bacteria, and is only produced in
response to infection. The infection thread grows through the root hair cells and often
penetrates other cells of the nearby root along the thread branches. The bacteria continue to
grow within the expanding network of tubes, produce nod factors that stimulate root cells to
proliferate, and eventually form a root nodule. Small nodules are visible to the naked eye
within a week of infection. Each root nodule is filled with thousands of living Rhizobium
bacteria, most of which are misshapen forms known as bacteroides. Parts of the plant cell
membrane surround the bacteroids. These structures, known as symbiosomes, can be
Containing several bacteroids or just one, they are sites where nitrogen is fixed.
Nitrogenase : An enzyme called nitrogenase converts nitrogen gas into ammonia in
nitrogen-fixing organisms. In legumes it occurs only within Bacteroides. The reaction requires
energy from ATP as well as hydrogen. The nitrogenase complex is sensitive to oxygen,
becoming inactivated when exposed to it. This is not a problem with free-living, anaerobic
nitrogen-fixing bacteria such as Clostridium. Free-living aerobic bacteria have different
mechanisms to protect the nitrogenase complex, including high metabolic rates and physical
barriers. Azotobacter overcomes this problem with the highest rate of respiration of any
organism, thus maintaining low levels of oxygen in its cells.
Rhizobium regulates oxygen levels in the nodule along with leghaemoglobin. This red, iron-
containing protein acts like hemoglobin. bound to oxygen. This provides sufficient oxygen for
the metabolic functions of the bacteroids but prevents the accumulation of free oxygen which
destroys nitrogenase activity. It is believed that leghaemoglobin is formed by the interaction of
plant and rhizobia as neither can produce it alone.