THE NITROGEN CYCLE

Nitrates are essential for plant growth

Root
uptake
Nitrate
NO
3
-

Plant
protein
2008 Paul Billiet ODWS
Nitrates are recycled via microbes
Nitrification
Nitrification
Ammonium NH
4
+

Ammonification
Nitrite NO
2
-

Soil organic nitrogen
Animal
protein
Root
uptake
Nitrate
NO
3
-

Plant
protein
2008 Paul Billiet ODWS
Ammonification
Nitrogen enters the soil through the
decomposition of protein in dead organic
matter
Amino acids + 1
1
/
2
O
2
CO
2
+ H
2
O + NH
3
+ 736kJ
This process liberates a lot of energy which
can be used by the saprotrophic microbes
2008 Paul Billiet ODWS
Nitrification
This involves two oxidation processes
The ammonia produced by ammonification is an
energy rich substrate for Nitrosomas bacteria
They oxidise it to nitrite:
NH
3
+ 1
1
/
2
O
2
NO
2
-
+ H
2
O + 276kJ

This in turn provides a substrate for Nitrobacter
bacteria oxidise the nitrite to nitrate:
NO
3
-
+
1
/
2
O
2
NO
3
-
+ 73 kJ

This energy is the only source of energy for
these prokaryotes
They are chemoautotrophs
2008 Paul Billiet ODWS
Root uptake
Nitrate NO
3
-

Plant
protein
Soil organic
nitrogen
Nitrogen from the atmosphere
Biological
fixation
Atmospheric
fixation
Out
gassin
g
Atmospheric Nitrogen
4 000 000 000 Gt
2008 Paul Billiet ODWS
Atmospheric nitrogen fixation
Electrical storms
Lightning provides sufficient energy to split
the nitrogen atoms of nitrogen gas,
Forming oxides of nitrogen NO
x
and NO
2

2008 Paul Billiet ODWS
Atmospheric Pollution
This also happens inside the internal combustion
engines of cars
The exhaust emissions of cars contribute a lot to
atmospheric pollution in the form of NO
x
These compounds form photochemical smogs
They are green house gases
They dissolve in rain to contribute to acid rain in the
form of nitric acid
The rain falling on soil and running into rivers
They contribute to the eutrophication of water
bodies
2008 Paul Billiet ODWS
Biological nitrogen fixation
Treatments Yield / g
Oats Peas
No nitrate & sterile soil 0.6 0.8
Nitrate added & sterile soil 12.0 12.9
No nitrate & non-sterile soil 0.7 16.4
Nitrate added & non-sterile soil 11.6 15.3
2008 Paul Billiet ODWS
Conclusion
Adding nitrate fertiliser clearly helps the growth of
both plants
The presence of microbes permits the peas to grow
much better than the oats
The peas grow better in the presence of the
microbes than they do with nitrate fertiliser added
The difference is due to the present of mutualistic
nitrogen fixing bacteria which live in the pea roots.
2008 Paul Billiet ODWS
University of Sydney
Alafalfa (Medicago sativa)
USDA - ARS
Root nodules
Only prokaryotes show nitrogen
fixation
These organisms possess the nif gene complex which make the
proteins, such as nitrogenase enzyme, used in nitrogen fixation
Nitrogenase is a metalloprotein, protein subunits being
combined with an iron, sulphur and molybdenum complex
The reaction involves splitting nitrogen gas molecules and adding
hydrogen to make ammonia

N
2
2N - 669 kJ
2N + 8H
+
NH
3
+ H
2
+ 54 kJ

This is extremely energy expensive requiring 16 ATP molecules
for each nitrogen molecule fixed
The microbes that can fix nitrogen need a good supply of energy
2008 Paul Billiet ODWS
The nitrogen fixers
Cyanobacteria are nitrogen fixers that also
fix carbon (these are photosynthetic)
Rhizobium bacteria are mutualistic with
certain plant species e.g. Legumes
They grow in root nodules
Azotobacter are bacteria associated with the
rooting zone (the rhizosphere) of plants in
grasslands
2008 Paul Billiet ODWS
Nitrate NO
3
-

Atmospheric
fixation
Out
gassin
g
Plant
protein
Atmospheric Nitrogen
Ammonium
NH
4
+

Soil organic
nitrogen
The human impact
Biological
fixation
Industrial
fixation
2008 Paul Billiet ODWS
Industrial N-Fixation
The Haber-Bosch Process
N
2
+ 3H
2
2NH
3
- 92kJ
The Haber process uses an iron catalyst
High temperatures (500C)
High pressures (250 atmospheres)
The energy require comes from burning fossil
fuels (coal, gas or oil)
Hydrogen is produced from natural gas
(methane) or other hydrocarbon
2008 Paul Billiet ODWS
The different sources of fixed nitrogen
Sources of fixed nitrogen Production / M tonnes a
-1

Biological 175
Industrial 50
Internal Combustion 20
Atmospheric 10
2008 Paul Billiet ODWS
Eutrophication
Nutrient enrichment of water bodies
Nitrates and ammonia are very soluble in
water
They are easily washed (leached) from free
draining soils
These soils tend to be deficient in nitrogen
When fertiliser is added to these soils it too
will be washed out into water bodies
There algae benefit from the extra nitrogen
This leads to a serious form of water pollution
2008 Paul Billiet ODWS
Fertilisers washed into river or lake
New limiting factor imposes itself
Sewage or
other organic
waste
Eutrophication
2008 Paul Billiet ODWS
Increased Biochemical
Oxygen Demand (BOD)
Hot water
from industry
(Thermal
pollution)
Pollution
from oil or
detergents
Reduction in dissolved O
2

Making things worse!
2008 Paul Billiet ODWS
The death of a lake
Death/emigration
of freshwater
fauna
Methaemoglobinaemia in infants
Stomach cancer link
(WHO limit for nitrates 10mg dm
-3
)
Increased nitrite
levels
NO
3
-
NO
2
-

Reduction in dissolved O
2

2008 Paul Billiet ODWS
The future of industrial nitrogen fixation
Food production relies heavily upon synthetic
fertilisers made by consuming a lot of fossil
energy
Food will become more expensive to produce
Nitrogen fixing microbes, using an enzyme
system, do the same process at standard
temperatures and pressures essentially using
solar energy
Answer: Genetically engineered biological
nitrogen fixation?
2008 Paul Billiet ODWS
Making things better
The need for synthetic fertilisers can be reduced by
cultural practices
Avoiding the use of soluble fertilisers in sandy (free
draining soil) prevents leaching
Rotating crops permits the soil to recover from
nitrogen hungry crops (e.g. wheat)
Adding a nitrogen fixing crop into the rotation cycle
Ploughing aerates the soil and reduces
denitrification
Draining water logged soil also helps reduce
denitrification
2008 Paul Billiet ODWS
Return to the atmosphere:
Denitrification
Nitrates and nitrites can be used a source of
oxygen for Pseudomonas bacteria
Favourable conditions: Cold waterlogged
(anaerobic) soils
2NO
3
-
3O
2
+ N
2
providing up to 2385kJ
2NO
2
-
2O
2
+ N
2

The liberated oxygen is used as an electron
acceptor in the processes that oxidise
organic molecules, such as glucose
These microbes are, therefore, heterotrophs
2008 Paul Billiet ODWS
Sediments 10 Gt
Nitrification
Root
uptake
Biological
fixation
Nitrification
Ammonium NH
4
+

Ammonification
Nitrite NO
2
-





Dissolved in water
6000 Gt
Denitrification
Leaching
Nitrate
NO
3
-

Soil organic nitrogen
9500 Gt
Atmospheric
fixation
Out
gassin
g
Industrial
fixation
Plant protein
3500 Gt
Animal
protein
Atmospheric Nitrogen
4 000 000 000 Gt
2008 Paul Billiet ODWS