Plant Physiology and Metabolism

Module 3
Transpiration
Academic Script

The loss of excess of water absorbed by the plant from its aerial parts in the form of
water vapours is called as transpiration. Transpiration from the leaf depends on two
major factors: (1) the difference in water vapor concentration between the leaf
air spaces and the external bulk air and (2) the diffusional resistance of this
pathway.

Kinds of transpiration:
There are three kinds of
transpiration.
1. Stomatal transpiration.
The transpiration that takes place
through the stomata is known as
stomatal transpiration. Most of the
transpiration takes place through
the stomata. The numbers of
stomata are more to the lower
surface of the leaves. In monocots they are equally distributed on all sides. In
aquatic plants with floating leaves they are present on the upper surface.
2. Cuticular transpiration or Peristomatal transpitation. Though cuticle is to
check transpiration still through cracks and thin layers transpiration takes place.
This trsnspiration is known as cuticular transpiration.
3. Lenticulr transpiration. Lenticels are the areas in the bark, which are filled
with loosely, arranged cells known as complimentary cells. The water vapour lost
through the lenticels are known as lenticular transpiration.
Significance.
Transpiration is regarded as boon to the plants for many reasons:
1) It create succession force and helps in the ascent of sap
2) It affects the DPD, thereby indirectly helping diffusion through the cells.
3) It affects the absorption of water and minerals by roots.
4) It helps in evaporating excess amount of water.
5) It also play a role in translocation of food from one portion of the plant to the
other.
6) It maintains a suitable temperature for the leaves.
7) It brings about the opening and closing of stomata which is indirectly influence
the process of photosynthesis and respiration.

Transpiration has some disadvantages too:

1) Transpiration consumes energy and cause unnecessary absorption of excess
water by roots.
2) When the rate of transpiration is high and soil is deficient of water an internal
water deficit is created which leads to wilting of plants.
3) Transpiration is the cause of death of millions of plants every year when it
exceeds the amount of water absorbed by the plant through roots.

In spite of these disadvantages the plant cannot avoid transpiration due to their
peculiar internal structure particularly those of leaves. So, many workers like Curtis
(1926) have called “transpiration as necessary evil.”
Factors affecting Rate of transpiration:

EXTERNAL FACTORS
1. Atmospheric humidity
When the relative humidity is high, the rate of transpiration decreases. It is because
the atmosphere more saturated with moisture and retards the diffusion of water
vapours from the intercellular space of the leaves to the outer atmosphere through
the stomata.
2. Temperature
An increase in temperature brings about an increase in the rate of transpiration by
lowering the relative humidity and opening the stomata widely.
3.Wind velocity
When the wind is stagnant the rate of transpiration remains normal.
When the wind is blowing gently the rate of transpiration increases because it
removes moisture from the vicinity of the transpiring parts of the plant, facilitating
more diffusion of water vapour from the from the leaves to the outer atmosphere.
When the wind is blowing violently the rate of transpiration is decreased because it
closes the stomata.
4. Light
Light increases the rate of transpiration because in light stomata opens and it also
increases the temperature.
5. CO2
An increase in concentration in the atmosphere over the usual concentration
inside the leaf retards transpiration because it leads to the closure of stomata.
6. Available soil water
Rate of transpiration will decrease if there is not enough water in the soil.
INTERNAL FACTORS
1. Stomatal frequency.
Stomatal frequency means the number of stomata per unit area of leaf surface. It
varies from plant to plant. If the stomata are open, with increased stomatal
frequency the rate of transpiration increases.
2. Structural peculiarities
Certain plants are adapted to reduce the rate of transpiration by reducing the size
of the leaves, forming sunken stomata, synthesizing thick cuticle etc. High osmotic
pressure of the leaf cells and presence of hydrophilic compounds such as gums,
mucilage, etc., help in retarding the rate of transpiration.
3. Other factors
Several other factors such as leaf area, amount of spongy tissue, volume of
intercellular spaces, orientation of leaf and extent of root system also effect the rate
of transpiration.

Structure of Stoma
Stomata are very minute openings or pores surrounded by two semi-lunar guard
cells. They occur only on the superficial layers i.e. in the epidermis of all plant
organs except in the roots. Each guard cell is either crescent shaped or dumble
shaped. The guard cells occur in pairs.Each pair is joined at the ends. The cell wall
of the guard cells possess cellulose microfibrils. These cellulose microfibrils are
oriented around the circumference of the elongated guard cells in a radiating
manner from the centre of the stomata. This microfibrial arrangement is called
radial micellation.
Source: Meidnerand Mansfield 1968.

Due to radial miscellation when a guard cell swells by absorbing water from the
surrounding cells it cannot increase in diameter but increase in length. As the two
guard cells are attached to each other at both ends they curve outward when the
guard cell expand. As a result the stomata opens.
A guard cell differs from epidermal cell in the following features:
1. The guard cell complexes have no direct connection with their adjoining tissues.
2. The guard cell contains few starch containing chloroplast.
3. Discrete ion adsorbent occurs beneath the poles of the guard cell pair in many
ferns and monocotyledons
Considering variable behaviour of stomatal movements, five categories have been
recognized:
1. Photoactive movements- light directly or indirectly controls stomatal
movement. Such stomata remain open during daytime and closed in the night.
2. Skotoactive movements- Stomata remain closed during daytime and open
during night. Such cases are found in succulent plant.
3. Hydroactive movement- In certain cases stomata close due to excessive loss
of water from the epidermal cells and open due to turgid condition of epidermal
cells. This is usually found during mid-day.
4. Autonomous movement- In some cases stomata open and close at a rate of
ten to fifteen minutes showing diurnal or rhythmic pulsation.
5. Passive and Active movement- Opening of stomata is considered as active
process while closing is the passive process and this is caused by the turgor
changes in the guard cells.

Mechanism of transpiration
Transpiration occurs in two stages:
(i) Evaporation of water from the cell walls into the intercellular spaces, and
(ii) Diffusion of these water vapours of the intercellular spaces into the outside
atmosphere.
In the first stage water absorbed through the roots and brought to the aerial parts
through ascent of sap gets evaporated from the surface of the turgid cells and
collects in the intercellular spaces increasing the water vapour pressure and
lowering the DPD. In the second stage this water diffuses through stomata, cuticle
or lenticels in the drier atmosphere outside because of low water vapour pressure
and high DPD value outside.

MECHANISM OF OPENING AND CLOSING OF STOMATA

The opening and closing of stomata depends upon the changes in the turgidity of
their guard cells. When guard cells are turgid, pores are open but when flaccid the
pores are closed. The size of the pore depends upon the degree of turgidity of
guard cells. Many theories have been proposed to explain the changes in the
turgidity of the guard cells.
1. Theory of Photosynthesis in guard cells
Since stomata opens in light and closes during night, Von Mohl (1856) proposed
that chloroplasts in the guard cells produce carbohydrates in the presence of light
by photosoynthesis. The photosynthates being osmotically active increase the
osmotic potential of the guard cell and subsequently the turgor pressure increases
and the stomata opens. This theory has lot of objections. Starch has been found in
very young guard cells even before it has emerged from the bud. Starch has also
been observed in guard cells of leaves kept in dark and also in albino leaves. Thus,
synthesis of starch (carbohydrate) in the guard cell is not dependent on
photosynthesis.
2. Theory of Starch – Sugar Interconversion
Sayre (1926) noted that stomata are sensitive to change in pH. A high pH favors
opening and a low pH favors closing of stomata. Yin and Tang obtained evidence
of the presence of phosphorylase in the chloroplasts that favors the degradation of
starch at pH 7.0 and favors starch synthesis at pH 5.0. Superficially, it seems that a
pH dependent phosphorylase regulates the starch sugar content and hence the
water movement into and out of the guard cells.
Thus the mechanism of opening and closing of stomata can be explained as : In
the morning due to photosynthesis carbon dioxide concentration lowers in the
mesophyll and guard cells. As a result pH of guard cells rises.
It stimulates enzyme phosphorylase. Phosphorylase converts starch into glucose 1-
phosphate. The latter is changed to glucose 6-phosphate which undergoes
hydrolysis to produce osmotically active glucose and phosphoric acid.

Evening closure of stomata is brought about by increased carbon dioxide content

(due to stoppage of photosynthesis) of leaf. It decreases pH of guard cells and
brings about phosphorylation of glucose. In the presence of phosphorylase,
glucose 1-phosphate is changed into starch.

As a result, osmotic concentration of guard cells falls. They lose water to adjacent
epidermal cells. With the loss of turgidity, the guard cells shrink and close the pore
in between them.
Objections:
(i) Glucose is not found in guard cells at the time of stomatal opening,
(ii) Starch ↔ Sugar changes are chemically slow while opening and closing of
stomata are quite rapid,
(iii) Wide changes in pH of guard cells
cannot be explained on the basis of
carbon dioxide concentration,
(iv) Onion and some of its relatives do
not possess starch or related
polysaccharide that can be hydrolysed
to the level of glucose,
(v) Blue light has been found to be more
effective than other wavelengths for
opening of stomata. The same cannot
be explained by starch hydrolysis
theory,
(vi) Hydrolysis of starch theory cannot account for high rise in osmotic pressure
found in guard cells.

Very small amounts of osmotically active sugars have been extracted from guard
cells. To date no explanation based on sugar build up in the guard cells adequately
account for guard cells turgidity change.

3. Theory of Proton Transport and Hormonal Regulation

Evidences indicates that the turgidity of guard cells of many species of plants is
regulated by K+, Cl-, H+ and malate.
When plants are exposed to light, the guard cells accumulate large amounts of K+.
The K+ accumulation seems to be due to the operation of an active exchange
process in which protons (H+) are pumped out of the guard cells into the
accessory cells and the K+ ions move from surrounding cells into the vacuoles of
the guard cells. The ATP required in ion
exchange process may come through
respiration. As the K+ ions migrate, they
are accompanied by Cl-. This anion
apparently moves in response to the
electrical differential created by the K+
uptake into the guard cell.
Potassium accumulation is also
accompanied by increase in pH, starch
degradation and organic acid mostly
malic acid build up. The organic anions
malate formed by the efflux of H+ ion are
neutralized by the influx of potassium. Thus in guard cells, osmotic and hence
water potentials become very negative due to the presence of K +, Cl, potassium
malate or di-potassium malate or both. A water potential gradient is established
from the accessory cells to the guard cell sap that results in water movement,
increased turgidity, and stomatal opening.
As a result of photosynthesis, CO2 concentration in guard cell decreases which
lead to increased pH in them during daylight. Organic acid chiefly malic acid is build
up during this period in the guard cells by the action of the enzyme PEP
carboxylase.
Closure mechanism involves participation
of an inhibitor hormone ABA which
functions in presence of CO2. In dark
ABA inhibits K+ uptake by changing the
diffusion and permeability of guard cells.
The K+ moves out the subsidiary cells.
ABA then induces the process of
acidification in guard cells that results in lowering of pH. At the low pH starch is
synthesized and thus osmotic pressure of the guard cells falls and water moves out
of guard cells to subsidiary cells. Guard cells then become flaccid and stomatal
pore is thus closed.
Source: the Mc.Graw-Hill Companies inc.

ROLE OF CO2 IN STOMATAL MOVEMENT

The concentration of CO2 in the substomatal cavities of a leaf influences stomatal
movement. It has been observed that if a leaf is placed in a stream of air free of
CO2, the stomata will open whether the leaf is in light or in darkness. On the other
hand if the leaf is exposed to concentrations of CO2 above the natural atmospheric
level (approx. 330 ppm), the stomata close even in the light.
The reason behind closing stomata is that CO2 dissipates the proton gradient
across protoplasmic membranes in guard cells. When the proton gradient is
dissipated, active transport of a potassium ion into guard cells ceases. Guard cell
dispose of accumulated ions. Potassium ion is transported out of guard cells into
subsidiary cells or other epidermal cells. The turgor pressure is reduced and the
stomata is closed.
The diurnal fluctuation of CO2 concentration in the leaf is due to the accumulation
of CO2 in the dark in absence of photosynthesis while the concentration of the
same is reduced due to photosynthesis in the light. Thus CO 2 act as a natural
agent for closing and opening of stomata.
In the leaves of CAM plants growing under conditions of water stress, malic acid
decarboxylation occurs through the day and internal tissue CO 2 concentration
increases dramatically. It is perhaps the high CO2 concentration in leaves of CAM
plant during the day time that explains their day time closure. Perhaps low CO 2
concentration in leaves at night accounts for nighttime opening.
 Role of Blue Light in Stomatal Movement
Blue light also stimulates stomatal opening through activation of proton pump in
guard cell plasma membrane. Blue light also stimulates starch hydrolysis and
malate biosynthesis.
The carotenoid zeaxanthin mediates the photoreception of blue light in guard cells.
It is synthesized in the carotenoid biosynthetic path way. It remains in the light
harvesting complexes of photosystem I and II. Signal transduction cascade for the
stomatal response to blue light starts with the excitation of zeaxanthin by blue
photons. The excitation is the isomerization of zeaxanthin. The isomerized
zeaxanthin cause a conformational change of an apoprotein. A second messenger
then transmits the signal across the chloroplast membrane. The transduced signal
led to the activation of the H+- ATPase at the plasma membrane. This is the
general process. Actually Calcium act as the second messenger. Ca2+- ATPase is
present in the chloroplast membrane. Zeaxanthin excitation by blue photons leads
to activation of Ca2+- ATPase at the chloroplast envelope. As a result it take up
calcium in the chloroplast. Low cytosolic Ca2+ activates the H+- ATPase in the cell
membrane.
In the morning stomatal opening is mediated by K + - malate, where as in the
afternoon it is mediated by sucrose accumulation due to starch hydrolysis or CO 2
fixation in the guard cells or uptake of apoplastic sucrose produced by mesophyll
photosynthesis.
Schematically it can be represented as:

Blue Excited zeaxanthin cause Ca2+- ATPase in the

Excitation of
light conformational change of chloroplast
zeaxanthin
an apoprotein membrane activated

Low cytosolic
Up take of calcium in
+
Eflux of H and Ca2+activates the H+-
the chloroplast
influx of K+ ATPase in the cell
membrane

Increase osmotic Opening

Transport of K+into the Development of
potential and water of turgor
vacuole to form K - malate uptake stomata
Measurement of transpiration
There several methods by which the amount of water vapor given off by plants can
either be estimated, or demonstrated. These methods may be grouped into three
classes.

1.In the first of these the plant is

potted, and weighed at intervals; the
loss of weight is assumed to be due
to the evaporation of water from its
surface.

2. In a second class the plant is

enclosed in a tight receptacle and the
vapor evolved from it is collected in
some kind of drying material which is
weighed before and after the
experiment, if there is any increase in
weight that is attributed to the absorption of water which was derived from the
plant.
3. In the third class the amount of water transpired is estimated from the amount
absorbed by the plant. However, the apparatus employed in this class is called
potometer.