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Biological Nitrogen fixation
Dr. Rachana Choudhary
(H.O.D.)Department Of Microbiology
Shri Shankaracharya Mahavidyalya,Junwani Bhilai
SYNOPSIS
1. INTRODUCTION
2. MODES OF NITROGEN FIXATION
3. ROOT NODULE FORMATION IN RHIZOBIUM-LEGUME
ASSOCIATION
4. ENZYMES OF NITROGEN FIXATION
5. MECHANISM OF NITROGEN FIXATION
6. BIOCHEMISTRY OF NITROGEN FIXATION IN NODULES
7. MOLECULAR BASIS OF NITROGEN FIXATION
8. CONCLUSION
9. REFERENCE
INTRODUCTION
The biological availability of N,P & K are
considerable an economic importance because
they are the major plant nutrients derived from th
e soil.
Of the three nitrogen is the most important
one this element is a key building block of protei
n molecule upon which life is based.
So the conversion of molecular nitrogen
into ammonia is known as Nitrogen Fixation.
Nitrogen is major
nutrient for plants
Atomospheric
N2
1.INTRODUCTION
Plant incapable to
Use this N2 gas
Triple bond makes
Highly inert
◆ Utilization of atmospheric nitrogen gas as a source of
cell nitrogen by way of its reduction to amonia is
called nitrogen fixation.
2. MODES OF NITROGEN FIXATION
• ( (a) Spontaneously nitrogen Fixation
(b) Industrial Process
(c) Biological Nitrogen Fixation
2. MODES OF NITROGEN FIXATION
(a) Spontaneously nitrogen Fixation
N2 H2
H2O
NH3NO2
NO3
(b) Industrial process
Fig: Production of ammonia by
HABER PROCESS
A large number of micro-organism are known to
have the ability to reduce atmospheric nitrogen
into nitrogenous compounds. This conversion of
molecular nitrogen into nitrogenous compounds
by micro-organism is called Biological Nitrogen
fixation
1.Non-symbiotic nitrogen fixation
2.Symbiotic nitrogen fixation
(c) Biological nitrogen fixation
1. Non-symbiotic nitrogen fixation
◆ Some microorganisms live freely and independently in
the soil.
◆ Including bacteria and some cyanobacteria.
FREE LIVING BACTERIA
Azotobactor,
Azomonas,
Azotococcus,
Thiobacillus ferooxidant,
Rhodospirillum rubrum
CYANOBACTERIA
Anabaena,
Nostoc
2. Symbiotic nitrogen fixation
■ Through nodules formation in legume plant:
■ Through Nodule formation in non-legume
plants
■ Without Nodulation
Legume
Plant
Association
Rhizobium
spp.
● Nodulating bacteria Rhizobium is gram negative, Motile rods.
● This association is fix larg quantity of nitrogen and have
considerable agricultural significance
■ Through nodules formation in legume plant:
Legume plant Nodulating bacteria
Alfalfa Sinorhizobium meliloti
Beans Rhizobium biovar
Clover R. trifolii
Lotus Mesorhizobium loti
Pae R.viciae
Soybean Bradyrhizobium japonicum
Through Nodule formation in non-legume plants
◆ Some non-legume plants involve in nodule formation.
◆ The best known plant is Alder associated with
actinomycete, Frankiya sp.
◆ Other plants are Casuarina and Hippophae.
Alder nodule Hippophae nodule
Without Nodulation
Example – Nostoc+Gunnera macrophylla.
Spirillum notatum+Maize
3. NODULES FORMATION IN RHIZOBIUM- LEGUME
ASSOCIATION
NODULES FORMATION TAKES PLACE BY
FOLLOWING STAGES :
(a) Recognition
(b) Attachment
(c) penetration
(d) Travel
(e) Bacteroid formation
(f) Development of mature nodule
(a) Recognition
◆ Legume plant secret
Metabolite including
Lectin protein.
◆ Which attracte Rhizobia
◆ Rhizobia comes in the
Rhizospheric region and
Increase population.
(b) Attachement
The bacterial
rhicadhesin
Attached with lactins.
(c) penetration
After attachement Rhizobia
Excrete nod factor which
Induce secretion of indol
acetic acid results
Curling of root hair.
(d) Travel
◆ After the root hair and
induce the Plant to develop
a cellulotic tube.
◆ Called infection thread.
(e) Bacteroid formation
◆ After releasing in cytoplasm
They became irregular shaped
Called Bacteroids.
◆ Surrounded by plant
Derived membrane called
Peribacteroid membrane.
(f) Development of mature nodule
◆ Symbiosome developed as
Nodule.
◆ The Symbiosome secret
Harmon called leghaemoglobin.
◆ which makes pink to nodule.
Vascular supply
To nodule
Rhizobia containing
Area in nodule
Formation of a Root Nodule
4. ENZYMES OF NITROGEN FIXATION
Nitrogenase
Dintrogenase
(Mo-Fe protein)
Dintrogenase reductase
(Fe-protein)
Mw= 220,000 Mw= 64, 000
Present in
Two copies
Present in
One copies
Alternative
Nitrogenase
◆ Nitrogenase that contain Vanadium in the place of
Molybdenum called alternative Nitrogenase.
◆ Synthesized if molybdenum is not present in medium.
Nitrogen fixation is catalysed by nitrogenase which is sensitive to
oxygen. It gets inactivated when the oxygen level exceeds 0.5 atm.
In plant cells, oxygen level is some what high, so the nitrogenase
may fail to reduced the nitrogen.
Leghaemoglobin is a red, myoglobin-like protein present only in
healthy root nodules of legumes. It is found outside the bacteria,
but in close contact with it.
Leghaemoglobin combines with oxygen to form
oxyleghaemoglobin and provides the oxygen to plant cells for
respiration. So the oxygen level around the bacteria is low & this
low content of oxygen does not affect nitrogenase activity for
nitrogen fixation.
In non legumin0us plant, where Leghaemo-
globin is absent hydrogenase enzyme is presen
t.
hydrogenase combines H2 and O2 to form wat
er (H2O). Hydrogenase removes oxygen from
the vicinity of nitrogenase in the bacteria.
Thus hydrogenase makes a suitable micro-envi
ronment for nitrogenase activity.
Azotobacter chroococcum &Anabaena cylindrica
contain the enzyme hydrogenase.
Hydrogenase
Factors Affecting Nitrogenase Activity
Excessive soil moisture,
Soil acidity,
P deficiency,
Excess mineral N, and
Deficiency of Ca, MO, CO and B.
Mechanism of biological nitrogen
fixation
5. MECHANISM OF NITROGEN FIXATION
6. BIOCHEMISTRY OF NITROGEN FIXATION IN ROOT
NODULES
Schematic presentation
Of biochemical
Pathway in nodule
Molecular Basis of Nitrogen Fixation
Nif Genes
The nif genes are genes encoding enzymes involved in
the fixation of atmospheric nitrogen in to a form of
nitrogen available to living Organisms.
Nif genes also encode a number of regulatory proteins in
volved in nitrogen fixation.
The nif genes are found in both free-living & sym-biotic
bacteria.
Conclusion
Biological Nitrogen Fixation is important
Process For the Increasing of soil Fertility .
It is necessary for the growth of the plant.
Agriculture in developed countries is definitely
base of the economy.
"eco- friendly“.
8. REFERENCE
◆ MICROBIOLOGY (Prescott)
◆ TEXT BOOK OF MICROBIOLOGY (R.P.Singh)
◆ INTRODUCTION TO EVS.MICROBIOLOGY (Michel.R.)
◆ CHEMISTRY (M.M.Tandan)
Biological Nitrogen fixation

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Biological Nitrogen fixation

  • 1. Biological Nitrogen fixation Dr. Rachana Choudhary (H.O.D.)Department Of Microbiology Shri Shankaracharya Mahavidyalya,Junwani Bhilai
  • 2. SYNOPSIS 1. INTRODUCTION 2. MODES OF NITROGEN FIXATION 3. ROOT NODULE FORMATION IN RHIZOBIUM-LEGUME ASSOCIATION 4. ENZYMES OF NITROGEN FIXATION 5. MECHANISM OF NITROGEN FIXATION 6. BIOCHEMISTRY OF NITROGEN FIXATION IN NODULES 7. MOLECULAR BASIS OF NITROGEN FIXATION 8. CONCLUSION 9. REFERENCE
  • 3. INTRODUCTION The biological availability of N,P & K are considerable an economic importance because they are the major plant nutrients derived from th e soil. Of the three nitrogen is the most important one this element is a key building block of protei n molecule upon which life is based. So the conversion of molecular nitrogen into ammonia is known as Nitrogen Fixation.
  • 4. Nitrogen is major nutrient for plants Atomospheric N2 1.INTRODUCTION Plant incapable to Use this N2 gas Triple bond makes Highly inert ◆ Utilization of atmospheric nitrogen gas as a source of cell nitrogen by way of its reduction to amonia is called nitrogen fixation.
  • 5. 2. MODES OF NITROGEN FIXATION • ( (a) Spontaneously nitrogen Fixation (b) Industrial Process (c) Biological Nitrogen Fixation
  • 6. 2. MODES OF NITROGEN FIXATION (a) Spontaneously nitrogen Fixation N2 H2 H2O NH3NO2 NO3
  • 7. (b) Industrial process Fig: Production of ammonia by HABER PROCESS
  • 8. A large number of micro-organism are known to have the ability to reduce atmospheric nitrogen into nitrogenous compounds. This conversion of molecular nitrogen into nitrogenous compounds by micro-organism is called Biological Nitrogen fixation 1.Non-symbiotic nitrogen fixation 2.Symbiotic nitrogen fixation (c) Biological nitrogen fixation
  • 9. 1. Non-symbiotic nitrogen fixation ◆ Some microorganisms live freely and independently in the soil. ◆ Including bacteria and some cyanobacteria. FREE LIVING BACTERIA Azotobactor, Azomonas, Azotococcus, Thiobacillus ferooxidant, Rhodospirillum rubrum CYANOBACTERIA Anabaena, Nostoc
  • 10. 2. Symbiotic nitrogen fixation ■ Through nodules formation in legume plant: ■ Through Nodule formation in non-legume plants ■ Without Nodulation
  • 11. Legume Plant Association Rhizobium spp. ● Nodulating bacteria Rhizobium is gram negative, Motile rods. ● This association is fix larg quantity of nitrogen and have considerable agricultural significance ■ Through nodules formation in legume plant:
  • 12. Legume plant Nodulating bacteria Alfalfa Sinorhizobium meliloti Beans Rhizobium biovar Clover R. trifolii Lotus Mesorhizobium loti Pae R.viciae Soybean Bradyrhizobium japonicum
  • 13. Through Nodule formation in non-legume plants ◆ Some non-legume plants involve in nodule formation. ◆ The best known plant is Alder associated with actinomycete, Frankiya sp. ◆ Other plants are Casuarina and Hippophae. Alder nodule Hippophae nodule
  • 14. Without Nodulation Example – Nostoc+Gunnera macrophylla. Spirillum notatum+Maize
  • 15. 3. NODULES FORMATION IN RHIZOBIUM- LEGUME ASSOCIATION NODULES FORMATION TAKES PLACE BY FOLLOWING STAGES : (a) Recognition (b) Attachment (c) penetration (d) Travel (e) Bacteroid formation (f) Development of mature nodule
  • 16. (a) Recognition ◆ Legume plant secret Metabolite including Lectin protein. ◆ Which attracte Rhizobia ◆ Rhizobia comes in the Rhizospheric region and Increase population.
  • 18. (c) penetration After attachement Rhizobia Excrete nod factor which Induce secretion of indol acetic acid results Curling of root hair.
  • 19. (d) Travel ◆ After the root hair and induce the Plant to develop a cellulotic tube. ◆ Called infection thread.
  • 21. ◆ After releasing in cytoplasm They became irregular shaped Called Bacteroids. ◆ Surrounded by plant Derived membrane called Peribacteroid membrane.
  • 22. (f) Development of mature nodule ◆ Symbiosome developed as Nodule. ◆ The Symbiosome secret Harmon called leghaemoglobin. ◆ which makes pink to nodule. Vascular supply To nodule Rhizobia containing Area in nodule
  • 23. Formation of a Root Nodule
  • 24. 4. ENZYMES OF NITROGEN FIXATION Nitrogenase Dintrogenase (Mo-Fe protein) Dintrogenase reductase (Fe-protein) Mw= 220,000 Mw= 64, 000 Present in Two copies Present in One copies
  • 25.
  • 26. Alternative Nitrogenase ◆ Nitrogenase that contain Vanadium in the place of Molybdenum called alternative Nitrogenase. ◆ Synthesized if molybdenum is not present in medium.
  • 27. Nitrogen fixation is catalysed by nitrogenase which is sensitive to oxygen. It gets inactivated when the oxygen level exceeds 0.5 atm. In plant cells, oxygen level is some what high, so the nitrogenase may fail to reduced the nitrogen. Leghaemoglobin is a red, myoglobin-like protein present only in healthy root nodules of legumes. It is found outside the bacteria, but in close contact with it. Leghaemoglobin combines with oxygen to form oxyleghaemoglobin and provides the oxygen to plant cells for respiration. So the oxygen level around the bacteria is low & this low content of oxygen does not affect nitrogenase activity for nitrogen fixation.
  • 28. In non legumin0us plant, where Leghaemo- globin is absent hydrogenase enzyme is presen t. hydrogenase combines H2 and O2 to form wat er (H2O). Hydrogenase removes oxygen from the vicinity of nitrogenase in the bacteria. Thus hydrogenase makes a suitable micro-envi ronment for nitrogenase activity. Azotobacter chroococcum &Anabaena cylindrica contain the enzyme hydrogenase.
  • 30. Factors Affecting Nitrogenase Activity Excessive soil moisture, Soil acidity, P deficiency, Excess mineral N, and Deficiency of Ca, MO, CO and B.
  • 31. Mechanism of biological nitrogen fixation
  • 32. 5. MECHANISM OF NITROGEN FIXATION
  • 33. 6. BIOCHEMISTRY OF NITROGEN FIXATION IN ROOT NODULES Schematic presentation Of biochemical Pathway in nodule
  • 34. Molecular Basis of Nitrogen Fixation Nif Genes The nif genes are genes encoding enzymes involved in the fixation of atmospheric nitrogen in to a form of nitrogen available to living Organisms. Nif genes also encode a number of regulatory proteins in volved in nitrogen fixation. The nif genes are found in both free-living & sym-biotic bacteria.
  • 35. Conclusion Biological Nitrogen Fixation is important Process For the Increasing of soil Fertility . It is necessary for the growth of the plant. Agriculture in developed countries is definitely base of the economy. "eco- friendly“.
  • 36. 8. REFERENCE ◆ MICROBIOLOGY (Prescott) ◆ TEXT BOOK OF MICROBIOLOGY (R.P.Singh) ◆ INTRODUCTION TO EVS.MICROBIOLOGY (Michel.R.) ◆ CHEMISTRY (M.M.Tandan)