Plant Ecology Lab: Environmental Correlates of Stomatal Density in Plants

Background:
As we’ve discussed in class, leaf stomata are the principal means of gas exchange in
vascular plants. Stomata are small pores, typically on the underside of leaves, which
are opened or closed under the control of a pair of banana-shaped cells called guard
cells.

When a plant has it’s stomata open, carbon dioxide can enter the leaf tissue for
photosynthesis. At the same time, water molecules evaporating from the stomata
pull water from the roots up through the xylem of the plant. While these processes
are fundamental to the survival of the plant, whenever the stomata are open, the
plant is losing water to the atmosphere. For plants native to places like California,
where there is an annual drought between April and October, water conservation is a
serious priority for plants, and keeping stomata open becomes costly. Plants have
acquired a variety of adaptations to help minimize the problems associated with
water loss through the stomata, one of which is to concentrate stomata on the lower
surface of the leaf, where the effects of temperature and wind on evaporation are
minimized. At the same time, the process of evaporation helps to cool the internal
tissues of the leaves (as sweating helps humans cool off). At high temperatures, the
proteins crucial to photosynthesis denature and lose their function, making it
beneficial for plants to open at least those stomata in the hottest parts of their
bodies. Clearly, plants face some trade-offs when it comes to the number, location,
and opening of their stomata, and through natural selection, these trade-offs are
reflected in the structure, physiology, ecology of different plant species.

Because of the critical role stomata play in the survival of plants, it has been
suggested that the density of stomata on the leaves of plants might be a useful
bioindicator of environmental change. In this lab you will propose a hypothesis about
how the density of stomata on a plant species of your choice relates to an
environmental variable that you think might be significant to the species you choose.
You will then test your hypothesis using observations of leaf imprints, and come to
some conclusion regarding your hypothesis,

Methods/Procedure:

1. Forming your hypothesis:

a. With your partners, go for a walk around campus, noticing factors that
could be related to stomatal density.
b. Find one type/species of plant that grows in two different
environmental conditions (i.e. wet versus dry, dark versus light), or for
which environmental conditions vary a lot from one part of the plant to
another (i.e. inner leaves versus outer leaves).
c. State a testable hypothesis relating stomata density and the
environmental factor you’ve chosen.
d. Bring a leaf sample and your written hypothesis to your teacher for
approval.
2. Collecting your specimens:
Once approved, select three leaves representing each environmental
condition (a total of 6 leaves). Remember that we want to control for
all variables other than the one we are testing, so try to choose leaves
that are as similar to each other as possible.
3. Making the imprints:
a. Back in the lab, paint a thick layer of clear nail polish on the underside
of each leaf. Be sure to keep the leaves representing each
 environmental condition separate – tape them onto a piece of paper to
make sure they don’t get moved or lost.
b. After the nail polish has dried (probably tomorrow), obtain a piece of
clear packing tape and put the sticky side of the tape against the nail
polish on your first leaf.
c. Gently pull the tape away from the leaf. You should see a cloudy
impression of the leaf on the tape.
d. Repeat for each leaf, again, be sure to keep track of which impression
goes with which leaf.
4. Observing stomata:
a. Tape each leaf impression to a clean microscope slide
b. Observe the slide under the high power objective of the microscope.
Move the slide around to get a sense of how consistent the stomatal
density is.
c. Pick a representative field of view and count the number of stomata
you see. Note how many are open versus closed (if you can tell).
d. Repeat for the remaining five leaves. Create a data table to organize
your results in.

Instructions for Formal Lab Report:

For this lab, you will present your results in standard scientific format. Here are some
brief instructions as to what to include in each section.

Introduction:

Give an overview of stomata, how they work, why they’re important etc. Include a
discussion of how stomata open and close, the mechanism by which guard cells pen
and close. Discuss the various trade-offs plants have to make regarding the number
and location of their stomata, and adaptations to different light/moisture/other
environmental factors shown by plants. Then explain the particular plant and
environmental factor that your group investigated, state and justify your hypothesis,
and explain what results would support your hypothesis,

Methods:

Very clearly explain what you and your partners did – what species/kind of plant (as
best we can figure), what the environmental factor you varied, etc. In this lab,
everyone will be doing slightly different things, so it’s important that you accurately
depict your methods so that someone else could replicate your work.

Results:

Present your data table along with one or two sentences that give an overview of the
data.

Discussion:

Do your data support your hypothesis? Explain. If you were going to continue this
study, what would you do differently, if anything? How many leaves do you think
you’d have to look at to get conclusive results? Were there complicating factors that
you couldn’t eliminate, or didn’t think about ahead of time? How might they have
influenced your results? Getting more general – how useful might stomatal density be
as a bioindicator for environmental changes (such as climate change?). Explain your
reasoning.