Lesson 4.

Nitrogen Nutrition
NITROGEN CYCLE
Essential element
Soils are usually deficient in N
NITROGEN CYCLE
ATMOSPHERE N2
DE
SN
IT
RI
FI

ON
CA
CI

TI
ÓN

XA
FI

NITROGEN CYCLE
SOIL

NH4+ NO2- NO3-

Nitrosomonas Nitrobacter
N
IO
AT
IC

NITRIFICATION
IF
N
O
M
AM

Organic N
ON
E R A LIZATI
MIN
 Cell
Plastid/chloroplast

NO3- NO3- NO2- NO2- NH4+

ATMOSPHERE
N2
DE
SN
IT
RI
FI
ON

CA
CI
TI

ÓN
XA
FI

SOIL

NH4+ NO2- NO3-

N

Nitrosomonas Nitrobacter
IO
AT
IC
IF

NITRIFICATION
N
O
M
AM

Organic N
TION
INE RALIZA
M
Nitrogen Nutrition

Inorganic N Organic N (“carbon skeleton”)

Stages: 1. ABSORPTION
2. REDUCTION (from NO3- to NH4+)
3. ASSIMILATION (NH4+ addition to carbonated skeletons) (aa)

SOIL PLANT
Proteins N2

DEGRADATION FIXATION

NH4+ carbonated
NO3- NO3- skeletons
ABSORPTION REDUCTION ASSIMILATION

NH4+
ABSORPTION Origin of NH4+
•N2 FIXATION
•NO3- REDUCTION
•NH4+ ABSORPTION
•PROTEIN CATABOLISM

Proteins aa Glutamate 2-Oxoglutarate

NH4+
1. NO3- ABSORPTION Nutrient absorption requires ATP and involves water absorption

ANIONS
H+ Pumos (ATPases) NO3-, PO4-, SO42-
CATIONS
Down their electric
Generate an CONTRANSPORT
gradient
electrochemical potential SYMPORT
difference (ANTIPORT)

Primary active transport Secondary active transport

1. NO3- ABSORPTION

• It is transported through the membrane by symport.

• It accumulates in the vacuole

H+ NO3-
Outside the cell (apoplast)

Plasma
membrane

Cytoplasm

ATP ADP + Pi
H+
 2. NO3- REDUCTION

Nitrate Nitrite
reductase reductase
NO3- NO2- NH4+

NADH + H+ Fdred + H+
NAD+ + H2O Fdox + H2O
Glycolysis/
Krebs
Cycle/
Photosynth
esys
Nitrate reductase: Cytoplasm
Nitrito reductasa: Leaves and roots plastids

Regulation of absorption and reduction of NO3- :

Light, nitrate
3. NH4+ ASSIMILATION Synthesis of aa

NH2 NH2
O

CH COOH NH3 ATP CH COOH C C OOH

CH2 Glutamine CH2 CH

2
Synthetase
CH2
CH2
H2O ADP + Pi CH2
C OOH

COOH C + 2 oxo glutarate

O NH2
GLUTAMIC Glutamine NH2
Synthesis of aa
ACID Glutamate
NH 2 synthase
CH COOH
NADPH
CH COOH Or Fdx CH2
(2e-)
CH 2
CH2
Glutámic Glutámic
CH 2
acid COOH acid
COOH
3. NH4+ ASSIMILATION

The transition from inorganic to organic N requires:

* ATP
Photosynthesis, glycolysis and respiration
* reducing power
* Carbon skeletons Oxidation of carbohydrates from
photosynthesis

N assimilation Carbohydrates metabolism

 N transport in the plant
Proteins Storage
proteins
SEEDS
NH4+ aa
NO2- aa
LEAVES

NO3-
NO3-
Vacuole
PHLOEM
XYLEM

Proteins aa
Age ROOT
NO3-
NO3- NO2- NH4+ Toxic!

NO3- NH4+
 N2
Nitrogen Fixation ATMOSPHERE DE
SN
IT
RI

ON
Nitrogen reservoirs: FI
CA

TI
CI

XA
● Atmosphere ÓN

FI
● Soil
SOIL
●N from biomass
NH4+ NO2-

N
NO3-

O
Nitrosomonas Nitrobacter

I
AT
IC
IF
NITRIFICATION

N
O
M
AM
Organic N
N
E RA LIZATIO
MIN

• Even though it comprises 80% of the atmosphere, plants and other eukaryotes cannot use
atmospheric nitrogen gas (N2)

• However, some bacteria are able to absorb N2 from the atmosphere and convert it to
ammonia in a process called nitrogen fixation
– Plants generally absorb nitrogen in the form of NO3- or NH4+ ions
 N2
Nitrogen Fixation ATMOSPHERE DE
SN
IT
RI

ON
FI
CA

TI
CI

XA
ÓN

FI
Nitrogen fixation requires a SOIL
series of specialized enzymes NH4+ NO2-

N
NO3-

O
and cofactors, including a large Nitrosomonas Nitrobacter

I
AT
IC
multi-enzyme complex called

IF
NITRIFICATION

N
NITROGENASE. The process

O
M
AM
is extremely energy demanding Organic N
N
E RA LIZATIO
MIN
• Requirements for the nitrogen fixation
– Expenditure of high-energy electrons
– 16 molecules of ATP per two molecules of NH3

N2 is reduced by microorganisms with the multi-enzyme complex called

NITROGENASE (exclusive from prokaryotes)

N2 + 16 ATP + 10 H+ + 8e- 2 NH4+ +H2 +16 ADP + 16Pi

Nitrogen Fixation The Role of Symbiotic Bacteria
Symbiotic N2 fixation. Non legumes

Nodule
Alder
Frankia

Actinomycete associated
with non legumes
angiosperms (Actinorhizal
plants)

Casuarina angustifolia Myrica gale

Nitrogen Fixation The Role of Symbiotic Bacteria
Symbiotic N2 fixation. Legumes

Rhizobium,
Bradyrhizobium Nodules
and other Rhizobia Soybean
Rhizobia provide the plant with ammonia in return for
sugar and protection Specificity

Legumes are one of the most important crops

Nitrogen fixation contributes to its production
They enrich the soil with N (cereal-legume rotations)
 Nitrogen Fixation The Role of Symbiotic Bacteria
Symbiotic N2 fixation. Legumes

NODULES
- Specialized organs
- Modified plant tissues
- Microorganisms are isolated
within the cells of the nodule and
within simbiosome (membrane
structure)
 Nitrogen Fixation The Role of Symbiotic Bacteria
Symbiotic N2 fixation. Legumes How Nitrogen-Fixing Bacteria
• The first infection event is recognition Infect Plant Roots?
– Recognition occurs between the root hairs of the plant and its symbiotic bacterium
• Young roots first release compounds called flavonoids
• When rhizobia contact the flavonoids, they produce Nod factors, which bind to the proteins on the membrane surface of the root-hair cells
NOD
Root hair FACTORS:
• Each legume species produces a
different flavonoid that acts as a
recognition signal, and each rhizobium
species responds with one or more 1º) Root hair
unique Nod factors curvature
• When Nod factors bind to the root-hair
surface, they set off a chain of events Infection
that leads to dramatic morphological Golgi thread
changes in the host legume vesicle

Golgi Body
The infection tube
FLAVONOIDS 2º) Cellulases membrane fuses with
the cell membrane
Transcription
NOD
NODFACTORS
GENES

GENES NOD
SIMBIOSOME:
functional
structure
where the N2
is fixed
Nitrogen Fixation The Role of Symbiotic Bacteria
Symbiotic N2 fixation. Legumes Root How Nitrogen-Fixing Bacteria
Infect Plant Roots?
1. Flavonoid released by root hairs attracts
rhizobia, which move into hairs
2. Rhizobia multiply inside the root hair,
resulting in the formation of an
infection thread
Root hair 1. Flavonoid released by
3. The infection thread grows into the root root hairs attracts rhizobia.
cortex Rhizobia move into hairs.
4. The infection thread buds off, releasing Flavonoid Rhizobia
rhizobia into the cortex cells Infection
5. The infected cortex cells divide rapidly, thread
2. Rhizobia proliferate
forming root nodules inside root hair and cause
an infection thread to form.

3. Infection thread grows

into the cortex of the root.

4. Infection thread buds

off, releasing rhizobia
Inside cortex cells.

Nodule
5. Nodule begins to
form from rapidly
dividing cortex cells.
ROOT
Absorbing organs

DICOT MONOCOT

Genetically Tª
Root shape Aeration
Edaphic environment
Mechanical impedance
Availability of H2O and Nutrients
 PLANT

RHIZOSPHERE

MICROORGANISMS SOIL
 BACTERIA
AMEBA
ROOT

NEMATODE

FUNGUS
 SOIL:
Physicochemical characteristics
(pH, texture, organic matter,
moisture, etc.)

Bacteria
Fungus
Protozoa
Bacteria
Nematodes
Fungus
Protozoa Microorganisms:
Nematodes
Hormones and growth regulators
Antibiotics
Plant:
Enzymes
Releases organic matter and
chemical signals that selectively Mobilize nutrients
attract microorganisms
Siderophores