PRACTICAL-2:

STRUCTURE AND DISTRIBUTION OF STOMATA

Stomata are tiny pores found in the epidermis of leaf. Size may be ranged from 7/3 µ
(Phaseolus vulgris) and 38×8 µ (in Avena sativa). Each stoma is normally surrounded by two
specialized eprdermal cells known as Guard cells. The guard cells are kidney shaped in
dicotyledonous plants and dumbbell shaped in the members of poaceae family
(monocotyledonous plants) and they remain joined at the end.

DUMBBELL SHAPED IN MONOCOTYLEDONOUS PLANTS

KIDNEY SHAPED IN DICOTYLEDONOUS PLANTS

Guard cells play significant role in opening and closing of stomata and the outer layer
guard cells wall is thin and elastic and inside the layer is thick and inelastic due to the
presence of a secondary layer of cellulose. Guard cells contains many organelles such as
nucleus, vacuole, chloroplast, starch molecules floating under cytoplasmic fluid, central
vacuole containing cell sap. In cytoplasm of guard cell, cytoplasmic streaming / motion is
remaining continued through which the transportation of nutrients, proteins, and organelles
occur within cells and helps in distribution of metabolic products.
 The epidermis layer of leaf contain numbers of stomata thousand and hundred
thousands per square centimeter (1000 — 60,000). The exact number depends upon the
spaces and the environmental conditions under which the leaf has developed stomata occur in
both the upper and lower epidermis of many plants. In general, they are commonly but not
always more abundant in the lower epidermis. In some others especially woody species they
are confined to the lower epidermis. In floating leaves, such as water lilies, lotus, stomata are
occurred only in the upper epidermis.

Stomata was discovered by Pfeffer & name ‘stomata’ was given by Malphigii.
Stomata cover 1-2% of leaf area. It is minute pore present in soft aerial parts of the plant.
Algae, fungi and submerged plants do not possess stomata.Stomata are minute pores of
eliptical shape, consists of two specialized epidermal cell called guard cells. The chloroplast
of guard cell are capable of very poor photosynthesis, because the absence of RUBISCO
enzyme.

In many gymnosperms and xerophytic plants (plants growing in desert), the stomata are
present embedded deeply in the leaves, so that they are not exposed to sunlight directly. Such
deeply embedded stomata are called sunken stomata. This is an adaptation to check excessive
transpiration in these plants.

METACALF AND CHALK RECOGNIZED FOUR TYPES OF STOMATA ON THE

BASIS OF THEIR STRUCTURE:

a. Anomocytic type:
In these stomata, accessory cells are absent. The guard cells are surrounded by ordinary
epidermal cells, e.g., families Ranunculaceae, Cucurbitaceae, Papaveraceae and Malvaceae.

b. Anisocytic type:
In these stomata the guard cells are surrounded by three accessory cells. Of these two are
larger whereas one is smaller in size.g., family Brassicaceae.
c. Diacytic type:
In these stomata the guard cells are surrounded by two accessory cells. Their common walls
are at right angle to the walls of guard cells, families Caryophyllaceae, Acanthaceae.
d. Paracytic type:
In these stomata the guard cells are also surrounded by two accessory cells, but their common
walls are parallel to guard cells, e.g., families Rubiaceae, Fabaceae etc.

TYPES OF STOMATA ON THE BASIS OF DEVELOPMENT (PANT, 1965):

1. Mesogynous type:
In this type of stomata guard cells as well as subsidiary or Accessory cells both are developed
from one mother cell. e.g. Rubiaceace &. Brassicaceae family.
2. Perigynous type:
In this type guard cells are formed from mother cell while subsidiary cells from nearby
mother cells, eg.: Cucurbitaceae family.
3. Mesoperigynous type:
In this type, guard cells & one subsidiary cells is formed from mother cell while other
subsidiary cells develop Independently. e.g.: Ranunculaceae, Caryophyllaceae family.

DISTRIBUTION AND TYPES OF STOMATA:

Depending upon the distribution and arrangement of stomata in the leaves five categories of
stomatal distribution have been recognized in plants.

1. Apple or mulberry (hypostomatic) type:

Stomata are found distributed only on the lower surface of leaves, e.g., apple, peach,
mulberry, walnut, etc.
2. Potato type:
Stomata are found distributed more on the lower surface and less on its upper surface, e.g.,
potato, cabbage, bean, tomato, pea, etc.
3. Oat (amphistomatic) type:
Stomata are found distributed equally upon the two surfaces, e.g. maize, oats, grasses, etc.
4. Water lily (epistomatic) type:
Stomata are found distributed only on the upper surface of leaf, e.g., water lily, Nymphaea
and many aquatic plants.
5. Potamogeton (astomatic) type:
Stomata are altogether absent or if present they are vestigeal. e.g., Potamogeton and
submerged aquatics.
 MECHANISM OF OPENING OF STOMATA:

 The guard cells having cell organelles such as nucleus, vacuole, poorly developed
chloroplasts (incapable of photosynthesis), starch molecules floating under cytoplasmic
fluid, central vacuole containing cell sap. They are surrounded by Subsidiary (epidermal)
cells.
 When turgidity increases, the outer thin walls of guard cells stretch outward thereby inner
wall is also stretching outward.
 The inner wall becomes concave and result is the space is occurring, pour widens and pore
opens. Stomatal Turgor Mechanism is involved in opening and closing of stomata.
 K+ (Potassium ions) play crucial role.
 The opening and closing of stomata are the result of active transport of K+ from epidermal
adjacent cells into the guard cells.
 Malic acid is synthesized from starch that excretion from chloroplast into cytoplasm of
guard cells in the presence of light.
Dissociation
Malic acid –R (COOH)2 Malate R (COO- )2 + 2H+
(Hydrogen ions)
Exchange
H+ K+ (adjacent cells)

 Influx of K+ and efflux of H+ that increase concentration of K+ in guard cell (Vacuoles)

thereby Osmotic pressure is developed; water is enter into the guard cells; increase turgor
pressure; guard cells becomes turgid and finally open the stomata.

MECHANISM OF CLOSING OF STOMATA:

 ABA play important role in closing of stomata.
 ABA inhibits K+ uptake by changing diffusion and permiability of guard cells. The K+
moves out to the subsidiary cells.
 ABA induces the process of acidification that lowering the pH. At the low pH, starch is
synthesized and osmotic pressure of guard cells falls and water moves out of guard cells to
subsidiary cells.
 Guard cells then become flaccid and stomatal pore is thus closed.

AIM:
To determine the stomatal frequency and stomatal index of some plants.

PRINCIPLE:
The number of stomata per unit area of the leaf is described as Stomatal frequency.

EXPERIMENT NO. 1:

To determine the phenomenon of transpiration:

MATERIAL REQUIRED:

A potted healthy plant, a bell jar which can cover the potted plant, a rubber sheet or oil cloth,
a glass sheet and grease are required for the experiment.

PROCEDURE:

 Cover the pot properly with rubber sheet or oil cloth leaving out the aerial parts of the
plant. This is done to prevent direct evaporation of water from soil surface and from the
surface of pot.
 Keep the pot on glass plate and covered air tight by the bell jar with grease on its rim.
Observations are made after sometime.
 Observe the water drops appear on the inner wall of the bell jar which comes from the
plant by phenomenon of transpiration.

EXPERIMENT 2:

To demonstrate continuity of intercellular spaces of a leaf and their relation to

stomata.

MATERIAL REQUIRED:
A wide-mouthed bottle with a cork having two bores, a leaf with broad lamina and long
petiole (Nymphaea) and aspirator

FIG.: DEMONSTRATE CONTINUITY OF INTERCELLULAR SPACES OF A LEAF

PROCEDURE:

 The bottle is filled more than half with water and its mouth is tightly closed with a cork
having two bores.
 In one bore, a glass tube is fixed and connected to an aspirator and in the other bore, the
cut end of petiole is introduced which dips in water.
 The apparatus is set airtight and the aspirator is set in action to suck air) from the bottle.
 With the start of aspirator, the chain of air bubbles begins emerging out from the cut end
of the petiole.
 The emergence a continuous stream of air bubbles reveals the fact that as only leaf
lamina is in direct communication with the external atmosphere, the air comes through it
via stomata and intercellular spaces. It is a clear-cut indication of continuity between
intercellular spaces and stomata and then between stomata and atmosphere.

Stomatal Index (I): It is defined as the per cent of stomata as compared to all the epidermal
cells (including the cells containing stomata) in a unit area of leaf.
 S
I = × 100
E+S
Where,
I = Stomatal Index, S = Number of stomata per unit area, T = Total number of epidermal cells
per unit area (No. of epidermal cell + No. of stomatal cells)