THE NITROGEN CYCLE

Nitrates are essential for plant growth

Plant
protein
Root
uptake

Nitrate
NO3-

2008 Paul Billiet ODWS

Nitrates are recycled via microbes

Animal
protein
Soil organic nitrogen
Ammonification

Plant
protein
Root
uptake

Ammonium NH4+
Nitrification

Nitrite NO2Nitrification

2008 Paul Billiet ODWS

Nitrate
NO3-

Ammonification

Nitrogen enters the soil through the

decomposition of protein in dead organic
matter
Amino acids + 11/2O2 CO2 + H2O + NH3

+ 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:

NH3 + 11/2O2 NO2- + H2O

+ 276kJ

This in turn provides a substrate for Nitrobacter

bacteria oxidise the nitrite to nitrate:
NO3- + 1/2O2 NO3-

+ 73 kJ

This energy is the only source of energy for

these prokaryotes
They are chemoautotrophs

2008 Paul Billiet ODWS

Nitrogen from the atmosphere

Atmospheric
fixation

Out
gassin
g

Atmospheric Nitrogen
4 000 000 000 Gt

Plant
protein
Biological
fixation

Soil organic
nitrogen

Root uptake

Nitrate NO3 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 NOx and NO2

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 NOx
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

No nitrate & sterile soil

Peas

0.6

0.8

Nitrate added & sterile soil

12.0

12.9

No nitrate & non-sterile soil

0.7

16.4

11.6

15.3

Nitrate added & non-sterile soil

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

Root nodules
Alafalfa (Medicago sativa)

USDA - ARS

University of Sydney

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

N2 2N
2N + 8H+ NH3 + H2

- 669 kJ
+ 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

The human impact

Atmospheric Nitrogen

Atmospheric
fixation

Out
gassin
g

Industrial
fixation
Plant
protein
Biological
fixation

Soil organic
nitrogen
Ammonium
NH4+

Nitrate NO3 2008 Paul Billiet ODWS

Industrial N-Fixation

The Haber-Bosch Process

N2 + 3H2 2NH3
- 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

Eutrophication
Fertilisers washed into river or lake

Sewage or
other organic
waste

2008 Paul Billiet ODWS

New limiting factor imposes itself

Making things worse!

Hot water
from industry
(Thermal
pollution)

Increased Biochemical
Oxygen Demand (BOD)

Reduction in dissolved O2

2008 Paul Billiet ODWS

Pollution
from oil or
detergents

The death of a lake

Reduction in dissolved O2

Increased nitrite
levels
NO3- NO2-

Death/emigration
of freshwater
fauna
2008 Paul Billiet ODWS

Methaemoglobinaemia in infants
Stomach cancer link
(WHO limit for nitrates 10mg dm-3)

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
2NO3- 3O2 + N2providing up to 2385kJ
2NO2- 2O2 + N2
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

Atmospheric Nitrogen
4 000 000 000 Gt

Atmospheric
fixation

Out
gassin
g

Industrial
fixation

Animal
protein

Plant protein
3500 Gt
Biological
fixation

Denitrification

Soil organic nitrogen

9500 Gt

Root
uptake

Ammonification

Dissolved in water
6000 Gt

Ammonium NH4+
Nitrification

Nitrite NO2Nitrification

Nitrate
NO3-

Leaching

Sediments 10 Gt
2008 Paul Billiet ODWS