Transpiration:

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2
 Transpiration

Transpiration is the evaporation of water from

plants. It occurs chiefly at the leaves while
their stomata are open for the passage of
CO2 and O2 during photosynthesis.

3
 Stomata

A stoma (plural stomata) is a tiny opening

or pore that is used for gas exchange.
They are mostly found on the under-
surface of plant leaves. Stomata open
and close to allow the intake of carbon
dioxide and the release of oxygen.

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 Importance of transpiration
Guard Cells What process involves
Guard Cells
What
using CO2 and H2O
goes releasing O2 as a waste
O2 H2O product?
out?
• Photosynthesis
What
goes CO2 What is the plant using this
in? process to make?
Stoma Closed
Stomata
Stomata Open • Carbohydrates-glucose

If the plant needs water for

photosynthesis, why is
water coming out of the 5
stoma?
 Stomatal transpiration

Cuticle
Prevents
water loss

Mesophyll
Site of
photosynthesis

Cuticle
Stomata Guard cells
Openings allow gases Open and close
and water to move in the stomata 6
and out of leaf
7
 Water transport in 3 parts

• Transpiration (or evapo-transpiration) is the

transport of water and minerals from roots to
leaves. It involves three basic steps:

– Absorption at the roots (STEP-1).

– Capillary action in the xylem vessels (STEP-2)..
– Evaporation at the leaf through stomata (STEP-3).

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1. Stomata are found in nearly all vascular plants
(except for submerged aquatic plants) Typical
stomatal density is 1000 - 100,000/cm2 of leaf
area.

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 Stomata: Characteristics
•Grasses and other monocots tend to have equal densities on
both surfaces.

•Herbaceous dicots have them on both surfaces, but greater

density on the lower (abaxial) side.

•Woody plants tend to have them only on the lower surface.

•Plants with leaves that float on water (e.g. water lily) have
them only on the upper (adaxial) surface.

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 Stomata: Structure

Stomata comprise a pair of highly specialized guard cells that

are encompassed by a pair of larger and thinner subsidiary
cells. In nearly all vascular plants, guard cells differ
significantly from other epidermal cells in having
chloroplasts. These differ from mesophyll chloroplasts in
lacking grana. Stomata also lack plasmodesmata, although a
full range of other subcellular organelles is present, including
nucleus, mitochondria, endoplasmic reticulum, Golgi
apparatus and ribosomes.

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Stomata: Structure

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 Stomata: Structure

Two distinct types of guard cells exist, kidney shaped (mostly

in dicot) and dumb-bell shaped (grassess). Kidney-shaped
guard cells are found in dicotyledons whereas dumb-bell-
shaped guard cells are found in grasses. Directly beneath
each pair of guard cells inside the leaf is a substomatal cavity.
Air in this cavity in living leaves is virtually saturated with water
vapour because of evaporation from adjacent wet cell walls.

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 Stomata: Structure
Kidney Shaped

14
Dumb Bell shaped
 Stomata: Structure
Cell walls of guard cells have two distinctive features: an
uneven thickening of walls forming the pore in either case
and a radial micellation of microfibrils. These two features
ensure that an uneven expansion occurs as guard cells
inflate. The two ends of the guard cells push against each
other to generate an opening. The thickened region
lining the edge of the pore cannot stretch lengthways
and therefore bends, generating an aperture. Guard
cells of grasses are more rigid with thickened regions
appressed when pores are closed. Inflating chambers at
each end force these sections apart while retaining
overall shape

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Stomata: Structure

16
 Guard cell properties and their relationship
with stomatal control

• Thickness of CW varies in the ventral and dorsal

part of the guard cells.
• Contains chloroplast and can perform light
reaction. (not dark reaction for the lack of key
enzymes)
• Structurally isolated from epidermal cells for the
lack of plasmodesmata (water and ions transmit
only through cellular pathway, thus helps to build
up water gradient) –No Symplastic flow
• Little volume, little amount of water absorption or
loss controls stomtal aperture.
• Glyoxalate cycle occurs 17
 How do stomata open and close?
1. Stomata open when water enters the guard cells from surrounding
epidermal cells.
2. This requires a water potential gradient
3. Guard cells are able to reduce their solute potential and thus their water
potential by increasing their solute concentrations.
4. This produces a water potential gradient so that water will enter them by
osmosis from the surrounding epidermal (subsidiary) cells.
5. As water enters, turgor increases, causing the guard cells to change
shape (become more curved).
6. As the cells become more curved, they push away from each other
causing the pore to enlarge.
7. The shape change happens because of radial micellation.
8. Cellulose microfibrils (micelles) are arranged radially

The mechanism namely the opening and closing of stomata depends upon
the turgor pressure in the guard cells. When the guard cells are turgid, the
stoma opens and when the guard cells lose water, stoma closes. 18
Guard Cell Function

Stomatal closing
1.Potassium ions move out of the vacuole and
out of the cells.
2.Water moves out of the vacuoles, following
potassium ions.
3.The guard cells shrink in size.
4.The stoma closes.

Stomatal opening
1.Potassium ions move into the vacuoles.
2.Water moves into the vacuoles, following
potassium ions.
3.The guard cells expand.
4.The stoma opens.

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 Mechanism of Stomatal Opening and Closing

The actual mechanism responsible for entry and exit of water to and from
the guard cells has been explained by several theories.
The most important theories are

i. The starch-sugar inter-conversion theory of Steward

ii. Active K+ transport of Raschke

iii. pH theory of Scarth

iv. Proton-potassium pump theory of Levitt.

 Mechanism of Stomatal Opening and Closing

Stomata opens: When potassium ion enters guard cells

Stomata closes: When potassium ion leaves guard cells

Why do stomata opens in day time?

Stomata has two sources of ATP production:

1. Oxidative phosphorylation (Mitochondria)
2. Cyclic photophosphorylation (Chloroplast) (no carbon fixation)

Day time stomata has high ATP content which drive H+-----K+ pump existing
between guard cells and subsidiary cells.

Thus H+ ion go out of guard cells to subsidiary cells

And K+ ion enters inside guard cells from subsidiary cells

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22
Due to influx of K+ ion and efflux of H+ ion pH of the guard cell increases (Ph 7.5)

At this high pH, Starch is converted into glucose (6C) inside guard celland
subsequently glucose is converted into Phosphoenol pyruvate (PEP, 3C) by
glycolysis

Glycolysis
Glucose (6C) PEP (3C)
pH 7.5

PEP carboxylase
PEP (3C) + CO2 OAA (4C)
Blue light Oxaloacetic acid
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 Reduction Malic acid (4C)
OAA (4C) + NADPH2
+ NADP (Guard cell)

Malic acid (4C) H+ + Malate-

(Guard cell to vacuole)

Thus H+ ion go out of guard cells to subsidiary cells

And K+ ion enters inside guard cells from subsidiary cells, enters inside
vacuole and then form potassium malate

Potasium malate is osmotically active, there by increase in OP of guard

cells in compared to subsidiary cells.

Water enters (from low OP to high OP) into guard cells from subsidiary
cells, there by TP develops, and stomata opens.

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What happen in dark

No cyclic photophosphorylation
Less ATP
H+---K+ pump stops

H+ enters guard cell and K+ ion leave guard cell by diffusion, without
using ATP

Due to entry of H+, pH of guard cell reduced

At low pH Potassium mallate dissociate into Go our side guard

cell by diffusion

K+ + Malate-
Potassium Mallate
(Guard cell)

Mallate combines with H+ to form

Mallic acid
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What happen in dark

Glyoxalate cycle
Mallic acid STARCH (Osmotically inert)

Low OP

So water move out of guard cells to

subsidiary cells

Guard cells become flacid, stomata

closes

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 Conditions which regulate stomatal opening
(1). Light
• Stomata opening are sensitive to red light and blue light, and blue light is more
effective, it stimulates opening by a blue-light receptor: zeaxanthin.

• Guard cell chloroplasts have a specific blue light response with an action
spectrum that resembles the action spectrum for blue light-stimulated stomatal
opening, suggesting a role of guard cell chloroplasts in the sensory
transduction of blue light. The xanthophyll, zeaxanthin has recently been
identified as a blue light photoreceptor in guard cells. The inhibitor of
zeaxanthin formation, dithiothreitol, inhibits zeaxanthin formation and the
stomatal response to blue light in a concentration-dependent fashion.

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Blue light-mediated stomatal opening

Ref: Sensory transduction of

blue light in guard cells.
Trends in Plant Sci 2000

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 Blue light-mediated stomatal opening

• Violaxanthin and zeaxanthin are the major constituents of the xanthophyll

cycle. The Arabidopsis npq1 mutant has a defective violaxanthin de-
epoxidase and cannot accumulate zeaxanthin. Zeaxanthin concentration
depends on the pH of the chloroplast lumen, which is modulated by rates of
electron transport at the thylakoid membrane and consumption of ATP and
NADPH by CO2 fixation in the Calvin cycle. The magnitude of the response to
blue light depends on zeaxanthin concentration and the number of absorbed
blue photons. The cascade is initiated by the excitation of zeaxanthin by blue
light, and the signal is transmitted to the cytoplasm, where it activates a
serine/threonine kinase. The kinase phosphorylates the C-terminus of the
H+ ATPase and activates the enzyme. A 14-3-3 protein binds to the
phosphorylated ATPase and stabilizes it. Dephosphorylation dissociates the
14-3-3 protein and deactivates the H+ ATPase

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 Blue/Red/Green light-mediated stomatal opening

Blue light

Trans Zeaxanthin Cis Zeaxanthin

Activates phot1/phot
2 receptors (14-3-3
protein)

PM-ATPase PM-ATPase
(Inactive) (active)

Efflux (outward movement) of H+ ion

Stomata Opens 30
 Blue/Red/Green light-mediated stomatal opening

Blue light Green light

Trans Zeaxanthin Cis Zeaxanthin

Inactivates phot1/phot
2 receptors (14-3-3
protein)

X PM-ATPase
PM-ATPase
(Inactive) (active)

Efflux (outward movement) of H+ ion

Stomata Close 31
 Blue/Red/Green light-mediated stomatal opening

Blue light Green light

Trans Zeaxanthin Cis Zeaxanthin

Inactivates Activates phot1/phot 2

phot1/phot 2 receptors (14-3-3
receptors (14-3-3 protein)
protein)

PM-ATPase
X PM-ATPase
(Inactive) (active)

Efflux (outward movement) of H+ ion

Stomata Close 32
Blue/Red/Green light-mediated stomatal opening

Red Blue

Phytochrome X Far red

PM-ATPase PM-ATPase
(Inactive) (active)

UV light Efflux (outward movement) of H+ ion

Stomata Close

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Blue/Red light regulate stomatal opening

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 (2) Temperature

• Stomatal aperture increase with Temp, within 20-

30℃ (the optimal).

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 (3). CO2

• Low CO2 conc. promotes stomatal opening, while

high CO2 conc. inhibits stomatal opening through
its acidification of the guard cell thus inhibits PM
hyperpolarization.

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 (4) Water content

• Stomata open when the leaf contain

enough water. When there is a water
shortage, they close.

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 (5) Plant hormones

• Cytokinin promotes opening

• ABA inhibits

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 ABA signaling in stomatal closing
1. During stress/ drought plant synthesizes ABA (Abscisic acid)
2. ABA binds to the ABR Receptor (ABAR) present in the plasma membrane of
Guard cells
3. Association of ABA with ABAR triggers activation of cell membrane signaling
molecule phospholipase C (PLC)
4. PLC activates intermediate signaling molecule IP3
5. IP3 interacts with Calcium (CA++) channels present on vacuoles to release
Ca++ ions in cytoplasm. It also causes more influx of Ca++ ions from
outside to inside guard cells
6. Enhance Ca++ ions inhibits K+ pump and inhibits the intake of K+ ions
7. Ca++ causes outward movement of Cl- ion from inside to outside of guard
cells
8. Ca++ close H+ pump, thus prevents out flux of H+ causing inside cytoplasm
acidic
9. As a results, outside of guard cells has K+ and Cl-, there by high solute
concentration, inner side of guard cell has low solute concentration. Due to
this osmotic imbalance water moves out from guard cells to surround cells,
resulting into closing of stomata

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ABA signaling in stomatal closing

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41
 Stomata: Growth Regulators

Presence of ABA (abscissic acid, a plant growth inhibiting hormone)

favours closing of stomata by blocking uptake of K+ by guard cells in the
dark. ABA act as stress hormone during drought condition.

Presence of Cytokinin (Plant growth regulator) is needed for the

active uptake of K+ ions

In the guard cells, the action of H+-ATPase activity is positively regulated by

blue light and auxins, whereas Ca2+ and ABA act as negative
regulators

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Hormonal crosstalk in the regulation of
stomatal closure and opening

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 From where water is transpired?
• Aerial parts of whole young plant
• Lenticels (lenticular transpiration) 0.1%
• Cutin (cuticular transpiration) 3%~10%
Stomatum (stomatal transpiration) ~ 90%

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 Transpiration: Water Transport

• Movement of water and minerals in a plant involves

entry into roots, xylem, and leaves.
• 3 processes:
1. Osmosis
2. Capillary Action (Adhesion)
3. Cohesion-Tension Theory

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 Water Transport
1. Osmosis - Water entering root cells creates
a positive pressure called root pressure.
a) Root pressure (primarily at night)
tends to push xylem sap upward in
plant.
b) Guttation is appearance of drops of
water along the edge of leaves, it is
result of root pressure.
➢ Root pressure is not a sufficient
mechanism for water to rise to the tops
of trees 46
47
 Water Transport
3. Cohesion-Tension Theory (Dixon & Jolly 1984)
a) Transpiration – evaporation of water from plants
b) Cohesion – water molecules attracted to other water molecules.
(polarity & hydrogen bonds)
c) Bulk Flow – water movement from roots to leaves as water
molecules evaporate from the leaf surface.

Cohesion tension theory is also known to us, as jolly's

theory. This theory describes the movement of water from
roots to leaves of a plant . Through osmosis water is carried
by xylem from root to leaves of a plant. Water molecules are
bonded to each other by hydrogen bond, hence water form a
string of molecules during its movement towards xylem . The
water molecules stick together and get pulled up by the
force called tension . This force is exerted because of the
evaporation at the surface of the leaf 48