THE NITROGEN CYCLE

© 2016 Paul Billiet ODWS

Nitrates are essential for plant growth

Plant
protein

Root
uptake

Nitrate
NO3-

© 2016 Paul Billiet ODWS

Nitrates are recycled via microbes

Animal
protein
Plant
Soil organic nitrogen protein

Ammonification Root
uptake
Ammonium NH4+

Nitrification

Nitrite NO2-
Nitrification
Nitrate
NO3-
© 2016 Paul Billiet ODWS
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.

© 2016 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 that oxidise the nitrite to nitrate:
NO2- + 1/2O2  NO3- + 73 kJ

 This energy is the only source of energy for these

prokaryotes
 They are chemoautotrophs.
© 2016 Paul Billiet ODWS
Nitrogen from the atmosphere
Atmospheric Atmospheric Nitrogen
Out fixation 4 000 000 000 Gt
gassing

Plant
protein
Biological Soil organic
fixation nitrogen Root uptake

Nitrate NO3-

© 2016 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

© 2016 Paul Billiet ODWS

Atmospheric Pollution
 This also happens inside the engines of cars
 The exhaust emissions of cars contribute to
atmospheric pollution in the form of NOx
 NOx form photochemical smogs
 NOx are green house gases
 NOx dissolve in rain to contribute to acid rain
in the form of nitric acid
 The rain falling on soil and running into rivers
 NOx contribute to the eutrophication of
water bodies.

© 2016 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

© 2016 Paul Billiet ODWS

Conclusion
 Adding nitrate fertiliser 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.

© 2016 Paul Billiet ODWS

Root nodules

Alafalfa (Medicago sativa)

USDA - ARS

University of Sydney
© 2016 Paul Billiet ODWS
Only prokaryotes show nitrogen
fixation
 These organisms possess the nif gene complex which
make the nitrogenase enzyme, used in nitrogen fixation
 The reaction involves splitting nitrogen gas molecules
and adding hydrogen to make ammonia

N2  2N - 669 kJ
2N + 8H+  NH3 + H2 + 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.

© 2016 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.

© 2016 Paul Billiet ODWS

The human impact
Atmospheric
fixation
Atmospheric Nitrogen

Out
gassin Industrial
g fixation
Plant
protein
Biological
fixation
Soil organic
nitrogen

Ammonium
NH4+

Nitrate NO3-

© 2016 Paul Billiet ODWS

Industrial N-Fixation
 The Haber-Bosch Process
N2 + 3H2  2NH3 - 92kJ
 The Haber process uses an iron catalyst
 High temperatures (500°C)
 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.
© 2016 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

© 2016 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.
© 2016 Paul Billiet ODWS
 Eutrophication
Fertilisers washed into river or lake

Sewage or New limiting factor imposes itself

other organic
waste

© 2016 Paul Billiet ODWS

Algal blooms can be natural

Annual cycles of an alga in the Baltic with Total Nitrogen and Total Phosphorus
© 2016 Paul Billiet ODWS
Controlling algal blooms

Lake Erie
 Limit nutrients
 Control pollution
 Increase number of
herbivores

© 2016 Paul Billiet ODWS

Making things worse!

Hot water Pollution

from industry from oil or
(Thermal detergents
Increased Biochemical
pollution) Oxygen Demand (BOD)

Reduction in dissolved O2

© 2016 Paul Billiet ODWS

The death of a lake

Reduction in dissolved O2

Increased nitrite
levels
NO3-  NO2-

Death/emigration
of freshwater Methaemoglobinaemia in infants
fauna Stomach cancer link
(WHO limit for nitrates 10mg dm-3)
© 2016 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?

© 2016 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.
© 2016 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 + N2providing 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
© 2016 Paul Billiet ODWS
Insectivorous plants

Drosera rotundifolia
© P Billiet
© 2016 Paul Billiet ODWS
Insectivorous plants
Drosera rotundifolia
© P Billiet

 Carnivorous plants trap

insects
 They digest their bodies
 Insect proteins provide a
source of nitrogen
 These plants have found
a niche in nitrogen
deficient soils
 E.g. peat bogs
© 2016 Paul Billiet ODWS
 Atmospheric Atmospheric Nitrogen
fixation
4 000 000 000 Gt
Out
gassin
g Industrial
fixation Animal
protein
Plant
protein
Biological Soil organic 3500 Gt Denitrification
fixation nitrogen 9500 Gt
Root
uptake
Ammonification
Dissolved in water
Ammonium NH4+ 6000 Gt

Nitrification

Nitrite NO2-
Nitrification

Nitrate
Leaching
NO3-

Sediments 10 Gt

© 2016 Paul Billiet ODWS