Assimilation of Mineral Nutrients

Nitrogen
Sulfur
Phosphate and cations
Nitrogen
Complex biogeochemical cycle
Extremely energy expensive process to assimilate
Atmospheric N≡N into ammonium (NH4+) costs 16 ATP per N
via bacterial nitrogen fixation
Nitrate (NO3- ) to Nitrite (NO2-) to NH4+ to glutamine requires
12 ATP/N
12.1 Nitrogen cycles through the atmosphere

Nitrogen assimilation
Nitrate uptake via low affinity and high affinity receptors in the root
Nitrate reductase uses reduced NAD(P)H to drive reduction of
NO3- to NO2-
Molybdenum complex, heme, and Flavin adenine
dinucleotide (FAD) domains
Nitrate reductase dimers use e- from NAD(P)H and protons to
reduce nitrate (NO3-) to nitrite (NO2-) + H2O

Nitrate reductase step

integrates various
factors
Light, nitrate concentration, and carbohydrate availability increase
NR transcription
Circadian regulation
Post-translational phosphorylation by a kinase recruits a 14-3-3
inhibitor protein that inactivates NR in darkness
Light, sugars, and nitrates dephosphorylate NR to restore
activity

12.4 Stimulation of nitrate reductase activity

Nitrite reductase
Nitrite is highly reactive
Reduction of NO2- to NH4+ occurs in plastids via nitrite reductase
using the reducing power of 6 reduced ferredoxins
Upregulated gene expression by light and nitrate
Downregulated transcriptionally by Gln and Asn

Nitrate assimilation
occurs in roots and
shoots
Varies enormously by species and environmental conditions
Nitrate is tolerated to high concentrations in plants, so can be
readily transported
12.6 Relative amounts of nitrate and other nitrogen compounds
in xylem sap

Ammonium metabolism
The Gln synthetase (GS) and Glu synthase (GOGAT) pathways
are present in cytosol and plastids, and regulated differently by
light and carbohydrates
Root GS forms glutamine for transport, root plastid GS as a
source of metabolic N amide substrates, chloroplast GS for
removing photorespiratory NH4+
Different GOGAT enzymes use NADH or Fdred
12.7 Structure and pathways of compounds involved in
ammonium metabolism (Part 1)

Figure 8.8 Operation of the C2 oxidative photosynthetic cycle

Ammonium scavenging as shown in the photorespiratory
pathway
-Preserves nitrogenous compounds
-Protects the cell from NH4+-induced proton gradient
abolition
Ammonium metabolism
The Glu dehydrogenase (GDH) pathway uses NAD(P)H in
mitochondria and chloroplasts to assimilate NH4+
Primarily for reallocation of N among different pools
13.7 Structure and pathways of compounds involved in
ammonium metabolism (Part 2)

Ammonium metabolism
Transamination reactions interconvert amine carriers for amino
acid and metabolic intermediate syntheses
12.7 Structure and pathways of compounds involved in
ammonium metabolism (Part 3)

Ammonium metabolism
Amino acids, especially Gln and Asn, are used for transport of
organic N compounds
Relative activities of Asn synthetase (AS) and GS/GOGAT
pathways balance N and C assimilation pathways
Abundant carbohydrates or light favor Gln (2N/5C; via
GS/GOGAT upregulation and AS inhibition) and thus rapid
incorporation into new growth
Limiting carbon or energy favor Asn (via AS) as storage and
transporter (2N/4C)
12.7 Structure and pathways of compounds involved in
ammonium metabolism (Part 4)

Amino acid metabolism

Unlike mammals, plants can synthesize all 20 essential amino
acids for protein synthesis
Carbons from glycolysis (3PG, PEP, pyruvate) or the citric acid
cycle (oxaloacetate,
α-ketoglutarate (2-oxoglutarate)) are transaminated from Gln or
Glu
13.8 Biosynthetic pathways for carbon skeletons of the 20
standard amino acids
Nitrogen Fixation
Major source of conversion from atmospheric N2 to NH4+
Fixation occurs in free-living bacteria (largely cyanobacteria) and
symbiotic bacteria
Symbioses may involve simple associations
Azolla and Anabaena
Apoplastic associations of N fixers with grasses
Complex associations
actinorhizal association of Alder with Frankia
legumes with rhizobia such as Rhizobium
Nitrogen fixation
Highly energy-rich intermediates are susceptible to damage from
O2
Nitrogenase enzyme must remain anaerobic
Heterocysts in cyanobacteria
 Root nodules

13.10 A heterocyst in a filament of the nitrogen-fixing

cyanobacterium Anabaena
Root nodules
Protect nitrogenase from oxygen
Physical barriers
Leghemoglobin carries respiratory O2 for bacteroid
respiration, accepting e- in a high-affinity oxidase at the end
of the e- transport chain
Allow ready access of carbohydrates to N fixers
Produced by complex “dance” of microbe and host plant
13.9 Root nodules on soybean

Root nodule formation

Host legume recognizes the bacterial
Nod factor via sugar-binding Lys-
motif receptors/kinases on the root
hairs
Ca2+ oscillations
Ca2+/Calmodulin-dependent protein
kinases (CaMK) promote transcription of
Nod factor-inducible genes
initiates infection and nodule
primordium formation
 Cytokinin-dependent inhibition of auxin
transport induces nodule cell division
and morphogenesis

Root nodule formation

Plant host nodulin (Nod) gene products and bacterial symbiont
nodulation (nod) gene products interact in signaling and
establishing nodules
Rhizobia migrate toward flavonoid chemical signals from
host roots
These signals induce rhizobial nodD activation, which
upregulates other nod genes
These nod genes code for enzymes that make lipochitin
oligosaccharide nod factors
Plant specificity comes from fatty acyl chain length and
saturation and modification of the chitin

13.11 Nod factors are lipochitin oligosaccharides

Root nodule formation

Rhizobial Nod factors induce nod-factor-inducible genes,
curling of root hair tip, enclosing bacteria
Localized degradation of cell wall allows bacteria-plant PM
contact
Plant PM invagination forms growing infection thread, while
root cortex cells divide and re-differentiate into a nodule
primordium
Infection thread full of rhizobia grows toward nodule
primordium
13.12 The infection process during nodule organogenesis (Part
1)

13.12 The infection process during nodule organogenesis (Part

2)

Root nodule formation

Infection thread full of rhizobia grows toward nodule primordium
Branching
Fusion with PM
Bacteria differentiate into bacteroids in the nodule, but
remain within plant PM-bounded vesicles (peribacteroid
membrane)
Nodule differentiates to minimize O2, and to extend
vascularization for fixed N export and C uptake
13.12 The infection process during nodule organogenesis (Part
3)

Nitrogenase complex
fixes N2
Fe and MoFe components catalyze:
N2 + 8e-+ 8H++ 16 ATP →
2NH3+ H2+ 16 ADP+ 16 Pi
Electrons come from reduced ferredoxin
O2 can damage both components of the complex
Nitrogenase makes up an enormous fraction of the total bacteroid
protein

13.13 The reaction catalyzed by nitrogenase

Fixed nitrogen is
exported in xylem
Form is species-dependent
Amides (e.g. Asn, Gln) in temperate-originating species
Ureides (allantoic acid, allantoin, citrulline) in tropical-origin
spp
Sulfur assimilation
-2
Primarily absorbed as sulfate (SO ) 4
+ -2
by root H - SO symporter, and
4
assimilated in the leaves.
SO4-2 is reduced to cysteine via
activation to adenosine-5’-
phosphosulfate (APS), then reduction
to sulfite (SO3-2) and then to sulfide
(S-2) for reaction with O-acetylserine
to produce Cys
Some Cys used to synthesize Met
and other derivatives
Export from the leaves as reduced
glutathione (GSH) or oxidized dimer
(GSSG)
13.15 Structure and pathways of compounds involved in sulfur
assimilation

Glutathione (GSH)
Three-amino acid peptide
Oxidized form is a dimer through a disulfide bridge between
cysteine residues (GSSG)
Phosphate assimilation
Primarily absorbed as phosphate (HPO4-2) by root H+- HPO4-2
symporter.
Assimilated into ATP and subsequently transferred to many
substrates
Cation assimilation
Coordination bonds are non-covalent associations of organic
carbon compounds with polyvalent cations
Cation loses positive charge through donation of unshared O
or N electrons from coordinating compounds
Electrostatic bonds are non-covalent attractions between positive
cation and negatively charged groups
Charges are maintained

Figure 13.16 Examples of coordination complexes

12.17 Examples of electrostatic (ionic) complexes

Iron assimilation
Iron is relatively insoluble in soil
Plants use:
soil acidification (root H+ export) to
increase Fe3+ and Pi solubility
reduction of ferric Fe3+ to more soluble
ferrous Fe2+ form, along with Fe2+
transporters
iron chelation with malate, citrate,
phenolics, and other compounds
Grass siderophores chelate Fe3+ and have siderophore-
Fe3+ transporters
3+
Transported as Fe -citrate complex
for insertion into heme precursors or
non-heme FeS proteins
Figure 13.18 Two processes through which plants roots absorb
iron
13.19 The ferrochelatase reaction

13.20 Examples of the two types of oxygenase reactions in cells

of higher plants (Part 1)
Oxygen assimilation:
-Mostly through respiration
-Dioxygenase reactions use both oxygens of O2 to
add to organic compounds
13.20 Examples of the two types of oxygenase reactions in cells
of higher plants (Part 3)
Monooxygenase reactions use one O from
molecular O2 for organic synthesis and the other for
water
Photoassimilation
Assimilation of nutrients is energetically expensive
N fixation can consume up to ¼ of the energy used in the
plant
In C3 plants, photoassimilation uses excess reducing power
and ATP from light reactions to assimilate N in chloroplasts
Abundant CO2 inhibits photoassimilation so reductant is
used in the carbon fixation cycle
Figure 13.20 Summary of the processes involved in the
assimilation of mineral nitrogen in the leaf