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Factors That AIIect The Number OI Stoma On A LeaI.

Stomata are specialised pors on the epidermis oI the leaves and stems oI plants, which
have the capability oI opening and closing. The opening and closing oI the stomata
are contolled by a pair oI modiIied epidermal cells called guard cells. The principle
Iuctions oI stomata are gas exchange Ior photosynthesis and the evaporation oI water
as part oI the transpiration stream.
The stomata open and close due to changes in turgor pressure within the guard cells.
In the centre oI each guard cell is a sap Iilled vacuole that can swell with increasing
turgor pressure. Guard cells increase their turgor pressure by accumulating K
thus lowering their water potential allowing an osmotic Ilow oI water into the cell.
Because the inner cell was is thicker and less elastic than the outerwall the cell
expnads unevenly. They bend and draw away Irom eachother. The result is the pore
between them opens.

Many Iactors acount Ior the extent to which the stomata open.
N Light levels-open during the day but closed at night.
N Hot temperatures-means an increase in the maount oI water lost through
evaporation. II the water loss exceeds the water uptake the water content oI the
plant Ialls and the guard cells lose their turgor and close.
N High levels oI CO
-in this situation the plant rewquires very little CO
so the
stomata close.

Similarly there are several Iactors that can aIIect the amount oI stomata present on the
epidermis oI leaves, including:
N :2/9
N Light levels
N #,31,
N Size oI the leaI
N Species oI the leaI.

In this investigation I will be determining the aIIect oI light levels on the
amount/density oI stomata.

I predict that the leaves taken Irom the outer area oI the shrub will have a
higher stomatal index than the leaves taken Irom the center oI the shrub.
Photosynthesis depends on the amount oI light absorbed by the photosynthetic parts
oI a plant. When chloroplasts in the leaI's cell are exposed to light they synthesise
ATP Irom ADP. Oxygen is produced as a by-product oI the photosynthesis reaction.
ThereIore increasing the concentration oI light will increase the amount oI ATP being
synthesised Irom ADP and so more oxygen will be released as a by-product. This
increase in photosynthesis will mean that larger amounts oI the other raw materials
are needed, carbon dioxide and water. Carbon dioxide enters the plant via diIIusion
through the stomata. II the overall rate oI photosynthesis is increased then a higher
concentration oI carbon dioxide must diIIuse into the plant. In order Ior this to happen
the plant was grow higher numbers oI stomta (creating a larger surIace area Ior CO

diIIusion. Similarly the large amounts oI oxygen being produced needs to be
excreted-this again occurs through the stomata. As the leaIs Irom the outer area oI the
plant are exposed to higher intensities oI light than the leaIs in the center oI the plant
they will have higher numbers oI stomata.
Higher intensities oI light would also increase the temperature to which the
surIace oI the plant is being exposed. The evaporation oI water through the stomata
has the aIIect oI cooling the plant. So iI the temperature is increased we can assume
that the number oI stomata will increase to enable more water to be evaporated, thus
cooling the plant.
However there has to be a balance. II the number oI stomata increase Ior gas
exchange this can lead to increased water loss. The plant must grow the correct
amount oI stomata to create a balance between the amount oI gas being exchanged
and the amount oI water being lost due to evaporation. For this reason I predict that
the diIIernece between the stomatal index on the outer most leaves and the inner
leaves will not be signiIicantly large.

In total twenty leaves will be taken, all Irom the same shrub/bush. Ten leaves
will be taken Irom the outer surIace i.e the area most exposed to light, and ten Irom
the centre oI the bush where light levels are low. The Iact that all the leaves will be
atken Irom the same plant ensures Iair testing, as there is a lot oI diversity between
diIIerent species oI plants.
The lower epidermis only (where the great majority oI the stomata are) oI each
leaI will be coated in a thin layer oI nail polish. AIter the nail polish has been leIt to
dry Ior a Iew minutes it will be peeled oII and placed onto a clear microscope slide.
The number oI stomata and epidermal cells will then be counted under high power
(400X). A stomatal index will be determined Ior the leaI. The Stomatal Index (I) ÷|S /
(E¹S)| * 100, where S is the number oI stomata per unit area, and E is the number oI
epidermal cells per same unit area. The data Iorm individual leaves will be combined
and a mean will be calculated Ior each plant sampled.
The slide will be looked at Irom three other random Iields oI view and the amount oI
stomata present in each Iield counted. To successIully count the number oI stomata in
a Iield oI view I will need to Iocus, using the Iine adjustment, up and down to bring
diIIerent planes into Iocus.
To ensure Iair testing:
N All the leaves must be taken Irom the same plant/bush/shrub to ensure there is
no diversity.
N All the leaves must be oI approximately the same size although as I will
determine the amount oI stomata per this is not essential. Similarly the
leaves must be oI about the same age, as those that are older are likely to have
more stomata than those that are younger, simply because they have been
growing Ior a longer period oI time.
N The same size area in which the stomatal index is calculated must be used Ior
each leaI.