Key Points Ecophys. Of Photosyn.

• RuBisCO slow & non-specific. O2 leads to wasteful PHOTORESPIRATION

• High temperatures, low CO2:O2 ration increase photorespiration
• C4, CAM & CCM minimize photorespiration by concentrating CO2 where
RuBisCO is active.
• C4 plants carry out C3 & C4 fixation in separate cells
• High CO2: O2 in bundle sheath (where C3 operates)
• CAM plants temporally separate CO2 uptake (night) from C3 (day)
• C4 & CAM carry an energy cost, but offer NET benefits in many environments
• This is reflected in their distribution
• C4 & CAM photosynthesis are examples of convergent evolution.
• C4 evolved in diverse Angiosperm groups.
• ~40% of Monocot species
• CAM also found in more primitive groups (e.g. Quillworts & Ferns) 1
 Key points: Water movement plants
• Water molecules are relatively cohesive, due to hydrogen bonds
• Water column ‘pulled’ up xylem under tension, not ‘pushed’ up from root.
• Rate of transpiration (demand) from leaf determines rate of transport (supply).
• The tension in the water column applies substantial stresses on the xylem
elements
• Xylem vessels & tracheids are reinforced by liginification.
• The water column in the xylem elements can break under tension
• Cavitation (embolism) can prevent or reduce xylem transport to shoot
• The phloem transports sugars and other products from source to sink tissues
• Source tissues (net producers) = mature photosynthetic organs (leaves)
• Sink tissues (net users) = roots, young leaves, flowers & developing seeds
• Bulk flow in the phloem is driven by:
• The loading of sugars into the sieve tube elements in source organs
2
• The removal of sugars from the sieve tube elements in sink organs
 BIO203 Lecture 6:
Secondary Growth & Nitrogen.

Instructor: Dr. Steven Chatfield

Office hours: Wed 1-2pm, Fri 2-3pm
Email: steven.chatfield@utoronto.ca
 BIO203: Plant form and function
• Reading:

• Mauseth Chapter 8 pages 170-191

As plants increase in height why don’t they
fall over?

Because they get wider too (duh!)

Plants increase in girth & undergo structural
reinforcement
• Plants can become woody with age,
strengthening the plant.

• Primary growth is from the shoot apex

and the root apex

• Increase in girth is due to a different

meristem: The Cambium

• This is a secondary meristem, giving rise

to secondary growth & a secondarily-
thickened stem
 Thick walled cells within xylem constitute
major proportion of structural reinforcement
Plants compensate for increase in size by adding
bulk and reinforcement to their stems
Strong cell types, sclerenchyma with
secondary walls composed of lignin:
tracheids, fibres, vessel elements
i.e. elements of xylem

Vessels do not
occur in conifer
wood
 Increasing the diameter of the plant
Secondary Growth occurs by the addition of new xylem (&
phloem) to the primary stem to make a secondarily-
thickened stem
secondary xylem = wood

woody dicots (trees, shrubs)

gymnosperms (conifers - pine, spruce)

requires cell division

meristem(s) adding to add girth by mostly lateral
(periclinal) divisions: lateral meristems
 Secondary growth
dicot primary stem

phloem
(pro)cambium

xylem

Initial growth is within the vascular bundles, by the pro-cambium:

the fascicular cambium.
Secondary growth

fascicular cambium

interfascicular cambium

Some ground tissue between the vascular bundles becomes

meristematic: interfascicular cambium.
Therefore a ring of cambium is formed
 The Cambial ring
Eventually the cambial cells (cambium) unite to a form ring
(cylinder) of meristematic tissue within stem
Radial growth
 Secondary growth

To add xylem or phloem, cambial cells divide in longitudinal

periclinal fashion: gives rise to either 1 phloem cell + 1 initial
OR
1 xylem cell + 1 initial

To add to vascular cambium (compensate for increasing girth)

initials divide anticlinally
Periclinal & anticlinal divisions
Cambium produces secondary xylem (wood)
to inside of this layer increasing girth

xylem

phloem cambium
1st
2nd 1 st year
3rd 3rd 2nd year
 Year 1
3 years of
secondary 1

growth

Year 2

3 2 1
1 2

Year 3
 Gymnosperm wood

Tracheids only

Resin ducts present (red arrow)

 From primary to
secondary growth Isolated procambial strands
 From primary to
secondary growth
Primary xylem and phloem

Cambial ring formation

 From primary to
secondary growth

Cambial ring and

secondary growth
 From primary to
secondary growth

Cork cambium & periderm

 Why are there annual rings?

Spring wood and summer wood. Poorer growth towards the end of
the growing season: smaller cells
Different types of vessel may be laid down at different times too
Rings more obvious in temperate/seasonal environments
Dendrochronology: Dating based on tree rings. Cross-referenced to other
methods (anchored to 11,000ya+). Map climate change
 Annual rings Dark wood.
Hard, very lignified

Slow growing.
Tight annual rings.
Very hard.
Fast growing. Wide annual
rings. Softwood. Gymnosperms
don’t always
make ‘soft’
wood
Figured wood
Heartwood & sapwood

Active and inactive

regions of the wood
Secondary growth in 3D
Secondary growth in 3D

Two cell types in vascular cambium

Fusiform initials - give rise to elements of

secondary xylem and phloem - the axial
transport system

Ray initials - give rise to rays - the radial

transport system

Multiseriate ray (several cells wide)

Uniseriate (one cell wide)
 Rays: Living parenchyma cells

Storage of nutrients
in winter

Allow passage of
Water/nutrients out
of the
xylem to other
regions
Lateral transport
Bark formation (cork & phloem)

xylem

phloem cambium
1st
2nd 1st year
3rd
3rd 2nd year
 Secondary phloem (inner bark)
• Much less of it than secondary xylem
• Typically contains many fibers
• Youngest cells are closest to the cambium, oldest become
crushed
• Only most recently formed cells are active
 Cork (outer bark)

Sugar maple Cork oak

Yellow birch
Cork formation

Cork (Phellem)
Cork cambium
(Phellogen)
Phelloderm*
Cortex
ExPhloem > Phloem

Vascular cambium
Xylem.
Cork formation

cork cells
cork cambium
phelloderm

Later in development new cork

cambium is often initiated below
existing cork layers to
compensate for increasing width
 Lenticels
The cork contains waxy substances (e.g. suberin) which make the bark
impermeable to water and gases.
To allow the outer cells to metabolize (they need oxygen) there are gaps
in the cork
They generally arise where
stomata were present on the
primary stem
 Monocots do not have 2ndary growth

The palm tree is a monocot

Primary monocot stem
Woody characteristic achieved by extensive primary thickening,
especially in the leaf bases
Phloem remains viable and active for longer than dicots
 Key points: Secondary growth
• Increase in girth of plant results for activity of secondary meristem
• The Cambium
• Dicots can become woody, xylem elements lignified
• Strengthens stems
• Steps in secondary growth:
• Procambium then Fascicular cambium forms between xylem & phloem of prmiary vascular
strands
• Meristematic cells arise in ground tissue between vascular strands giving rise to interfascicular
cambium
• Cambium unites to form ring of meristematic tissue in stem
• Xylem cells are added to inside of ring or phloem cells to outside by periclinal cell divisions of
cambial initials
• To compensate for increasing girth new initials are produced by anticlinal divisions
• Two types of cells in vascular cambium, fusiform & ray initials
• Fusiform ->xylem & phloem; Ray initials -> radial transport system
• Reaction wood. Tension wood in dicots; compression wood in gymnosperms
• 2dary phloem (less than xylem), only youngest cells functional rest crushed
• Cork cambium arises below epidermis & gives rise to outer bark
• ‘Woody’ monocots utilize extensive primary thickening (e.g. of leaf bases)
 BIO203:
Nitrogen.

Instructor: Dr. Steven Chatfield

Office hours: Wed 1-2pm, Fri 2-3pm
Email: steven.chatfield@utoronto.ca
 BIO203: Plant form and function
• Last Reading:

• Mauseth 6th edition Chapter 13 pages 341-363

• Mauseth 5th edition Chapter 13 pages 313-332
 Nitrogen
• Acquiring and using N

• Why is this important?

– It’s essential!
• Agriculture, food production
and supply
– We use 10kcal for every 1kcal
we get
– Fossil fuels & Fertilizer
 Nitrogen is Essential
• N atom can form up to 3 covalent bonds
– It is relatively electronegative (3) & will form
polar bonds with H
– Important because it is less electronegative
than O & H-bonds it participates in are
weaker & easily separated, e.g. DNA strands
Nitrogen is found in most complex
molecules of life, including:
– All amino acids
• Major demand = RuBisCo (up to 50%
leaf protein in dicots).
– All nucleotides
– Pigments e.g. chlorophyll
– Many plant hormones
– Secondary metabolites
 Nitrogen
• N2 Gas = 78+% of the atmosphere
• N often the most limiting nutrient in temperate systems
– Plants can only take up & use N in certain forms
– These useable forms of N interact with soil particles
• N N bond very stable & this form of N is inaccessible to
plants
– Only certain bacteria can break this bond
• N-H & N-O bonds require much less energy to break
– Wide variety enzymes can utilize molecules with these bonds &
incorporate N into different biological molecules
 Available Nitrogen
• NH3, NH4+ = ammonia, ammonium. NH4+ = usable
form that can be taken up by plants
• NO2-, NO3- = nitrite, nitrate = usable form that can
be taken up by plants
– Nitrate is the form taken up most by plants
– Ammonia rapidly converted to nitrate by soil
bacteria
• Therefore, triple N2 bond must be broken & the molecule
reduced using electrons to form N-H or N-O bonds.
 Available Nitrogen
• Nitrite & Nitrate are both
negatively charged.
• In temperate soils the majority NO3-
of soil particles are also
negatively charged.
– Not well retained by the soil
• Easily washed down the soil
horizons & out into ground
water & water courses.
– They are LEACHED, especially
after rainfall.
 Available Nitrogen
• Nitrite & Nitrate are both negatively
charged.

• In temperate soils the majority of soil

particles are also negatively charged.
– Not well retained by the soil

• They are relatively easily washed down the

soil horizons & out into ground water &
water courses.
– They are LEACHED, especially after rainfall.
 Getting useable N from N2 gas
• Nitrogenase (complex), can convert inorganic N2 gas into
organic nitrogen, ammonia (NH3), It can break the triple bond in
N2 gas
• O2 inhibits nitrogenase activity
– Needs anaerobic conditions
Some Bacteria (Nitrogen-
Fixing Bacteria)
That evolved in an
anaerobic environment
possess nitrogenase

Plants do not
 Plant strategies for getting N: 1 Attract free-
living microbes
• Plants attract a free-living microbes to build a
“rhizosphere" around their roots.
• 15% of photosynthate is deposited in the soil
Carbohydrates from sloughed-off
root cap cells, slime (mucilage)

Supports bacterial growth

Bacterially generated N made

available to plant by amoebae & fungi
feeding on bacteria & organic matter From: Plants, Genes & Biotechnology
Chrispeels & Sadava
 Plant strategies for getting N: 2 Maximize
surface area for uptake
• To maximize contact between roots & sources
of N, plant roots need a large surface area for
uptake
• Lateral root outgrowth is often responsive to
Nitrate or Ammonium
– Nitrate diffusible & signals nutrient concentrations
• Roots grow continuously.
– The N uptake from soil around a section of root stops 5-6
days after the region 1st penetrated.
– Roots are a huge drain on the C/energy output of
photosynthesis
• Soil aeration is important to support this
growth. sugars are used for respiration which
requires O2 & gives off CO2
 Plant strategies for getting N: 2 Maximize
surface area for uptake
• The surface area for uptake of
water, N & other nutrients is
enormously expanded by root
hairs.
• Each root hair is an outgrowth
from a single epidermal cell
• May be mm-cm long
• Most water & nutrients
including N taken up across
their surface From: Plants, Genes & Agriculture. Chrispeels & Sadava
 Root hairs and nutrient uptake
• The surface area for
uptake of water, N & other
nutrients is enormously
expanded by root hairs.
• Cell membrane of root hair
covered in uptake
transporters for nutrients
incl. nitrate
• High affinity & low affinity
uptake carriers
• Energy costs?
 Plant strategies for getting N: 3 Symbiotic
associations with fungi
• MYCORRHIZAE (most plants can form these)
• Fungi use extensive hyphae to scavenge nutrients (particularly
important for N & P) over a much Greater surface area/volume of soil
than the root

Plant gets N + P, fungus gets fixed C

Some grow over surface of root (ECTO) & some
grow into it (ENDO or VAM) (may even ‘enter’
cells of the root = VAM)
Can greatly increase plant growth,
particularly on nutrient poor soils (Wheat
220% Corn 122% & Onion 3155%)
May also help protect against some pathogens &
parasites
From: Plants, Genes & Biotechnology Chrispeels & Sadava
 Plant strategies for getting N: 4 Legume-
Rhizobium symbiosis
• Symbiotic relationship between N-fixing bacteria (Rhizobium) &
“legume” plants. Add 50-200kg N/ha
• In soybeans even on good soil up to 1/3 of their N comes from
symbiosis
Nodule provides an anaerobic environment in
which bacteria can Fix N
Bacteroids (Fill Cell)
Leghaemoglobin (Pink Nodules)
N fixation needs reduced Oxygen

Nodules = complex structure with

‘meristematic’ region & vascular connection
Nodules take time to develop
Seedlings still dependent on N
stored in seed + soil N
 Plant strategies for getting N: 4 Legume-
Rhizobium symbiosis
• Formation of nodule requires
coordinated exchange of signals
between the plant & the bacteria
• Plant responds to bacteria
Infection Thread
signals for changes in root hair, Delivers Bacteria To
passage of the “infection Root Cortex Cells

thread” & coordinated gene

expression to maintain &
provision the nodule
• Bacteria must respond to
signals from specific species of
legume & coordinate its gene
expression
– NOD genes
Plant strategies for getting N: 4 Legume-
Rhizobium symbiosis
 Legume-Rhizobium symbiosis: Costs and
benefits
• Gets N in organic form
• Gives fixed carbon to bacterium
• Energetically expensive for
nitrogenase to break N2 & produce
NH3
– 16 ATP for each N2
– ~20% of photosynthate produced by
legumes used for N fixation
• Gets fixed carbon
– Bacteroids grow and multiply
– Uses energy to fix N
• Fixes N and gives to plant
• Bacteroids die when nodule
does
– Chemicals secreted by nodule
help free-living forms
 Non-Legume nodules
• Parasponia (same Family as Cannabis)
– Nodules with Rhizobia
• Alder & other actinorhizal plants
– Nodules with other bacteria
• E.g. Frankia
 Other N fixing relationships?
• Endophytic bacteria
– Not as efficient as Rhizobium-Legume
symbiosis
– Often in root (lower O2?), but also found in
stems
– Grasses: Sugar cane, Maize, Wheat, Rice
– Poplar & Willow
– Sequenced: Azolla
• Klebsiella pneumoniae 342
• Pseudomonas stutzeri A1501
• Azoarcus BH72
• Cyanobacteria
– Gunnera plant
– Cycads Anabeana
– Azolla – Anabeana in rice paddies
Plant strategies for getting N: 5 Murder & theft
• Carnivorous plants trap and
digest insect pray (mostly) as
sources of N (and other nutrients)
– Often live in low N availability
environments
• Sundew & Venus flytrap (bogs)
• Pitcher plants (bogs, epiphytic)
• Parasitic plants (e.g. Monotropa)
may tap into their hosts N
reserves
– Directly or indirectly..
 Plant strategies for getting N: 6 become a
toilet for small mammals
• Pitcher plant from Borneo
– Nepenthes rajah
– Nepenthes lowii
 Synthetic source of N
• HABER-BOSCH process = A
synthetic way to break the N2
triple bond
• Requires lots of energy
– 450-600 C + 100s ATM
• Explosives & fertilizer without
depending on Guano
• Costs, but fertilizer has a strong
+ve impact on yield
• Leaching leads to pollution of
ground water Fertilizer use may favour micro-
– Blue-baby syndrome organisms that utilise nitrate &
– Toxic algal blooms transform it back to N2
 N: Uptake and assimilation
Nitrogen Assimilation
NO3- transported to shoot by
transpiration stream (xylem)

Converted to NO2- in
cytoplasm

NO2- → chloroplast
NO2- → NH4+
NH4+ used to create
glutamine from glutamate
 Transport & storage of N
• Transport forms = Amino
acids (phloem)
– Soluble building blocks of
proteins
• Storage proteins
– Prolamins, globulins inside
organelles (protein storage
bodies)
• Dicots = PSBs mostly in
cotyledons
• Monocots = PSBs in
endosperm of seed
– Corn storage protein = Zein
– Wheat storage protein = Gluten
 Control & regulation of plant N
• Nitrogen energetically expensive to fix by legumes
– 12 - 17g carbohydrates consumed per 1g N fixed by
Rhizobium

• NH4+/NO3- is energetically expensive to uptake from soil &

assimilate into amino acids
• To assimilate N into an amino acid, must also be enough
‘C- backbones’

• 50-60% of leaf N needed for Rubisco & other

photosynthetic enzymes
 Control & regulation of plant N
• Light & sucrose up-regulate N assimilation
• Darkness + sucrose repress N assimilation
• Repression of N assimilation when organic N is high
relative to C
• Uptake also regulated
• Shoot/root ratio also responsive & helps balance N uptake
against photosynthesis
• When organic N high relative to C, then the N is shunted to
storage as the amino-acid Asparagine
– Has higher N:C ratio than Glutamine & stores organic N
more efficiently when C- backbones limited
 C & N use Summary
• Molecules that store both N & C or need both elements for
their production
– Amino acids & proteins (including storage proteins)
– Nucleotides, DNA & RNA
– ATP, NAD(H) & NADP(H) (cofactors)
– Pigments (e.g. chlorophyll)
– Many secondary metabolites & plant hormones

• Molecules that do NOT require, or store, N

– Fats & oils
– Carbohydrates: sugars, starch & cellulose
 Nitrogen: Key points
• Plants have a High Requirement for Nitrogen, but Cannot Use Atmospheric N2 (Strong
Triple Bond). Need NO3- or NH4+
– NO3- tends to be leached because soil particles in temperate soils are usually negatively charged
• Root Structure (Hairs & Branching) & Continuous Growth Help Exploit Soil Organic N
Sources.
• Symbiotic Relationships Used to Obtain N in Exchange for Fixed C. Signaling Between
Partners
– Associations With Fungal Partners to Form Mycorrhizae to Scavenge N (+ Water & P) from
Environment (Most Plants)
– Associations Between Legume Plants & N Fixing Bacteria Which Convert N2 to NH3 Using the
Enzyme Nitrogenase in the Tailored Anaerobic Environment of Nodule
• In low nutrient environments plants may resort to capturing small animals
• N2 Triple Bond can be Broken By High Energy Haber Process to Produce Inorganic Fertilizer.
Alternative Strategies Include ‘BioFertilizer’ or Utilizing N-Fixation (Legumes-Rhizobium, Azolla-
Anabaena)
• NO3- Primary Uptake Source. Via Special Transporters. To Shoot Via Xylem
– NO3- Converted to NO2- in Cytoplasm NO2- → Chloroplast NO2- → NH4+
– NH4+ Used to Create Glutamine from Glutamate
• Thereafter, N is Transported as Amino Acids (Phloem) & Stored as Proteins
• C Fixation vs N Uptake & Assimilation (& Fixation in Nodules) tightly regulated