BIOL 1507E Leaves

Summary
A. The Mesomorphic Leaf
1. The eudicotyledon leaf
2. Stomata count
3. The high-efficiency leaf
B. The Xeromorphic Leaf
C. The Hydromorphic Leaf

Even though land plants are well adapted to life out of water, their
photosynthesizing cells still require an aquatic medium. This is a holdover from ancestral
green algae in which photosynthetic cells were in direct contact with water. The aquatic
medium is required because the CO2 necessary for photosynthesis is found in the cell not
as a free gas, but dissolved in water. This has resulted in the land plants evolving a leaf
structure that contains a watery “culture” of chlorenchyma (cells with chloroplasts)
sandwiched between two protective layers, the upper and the lower epidermis.

The epidermis is porous to allow passage of carbon dioxide and oxygen. Water is
unavoidably lost through these pores, and the potential exists, therefore, for the “culture”
to dry out.

In this exercise you will see how the internal features and epidermis of the leaf
help to abate moisture stress. You will study the photosynthetic tissues and the ways they
are modified to receive light and maintain a saturated internal atmosphere.

A. The Mesomorphic Leaf

The majority of flowering plants are mesophytes, that is, plants requiring
moderate amounts of water regularly throughout the growing season. Their leaves are
described as mesomorphic.

1. The eudicotyledon leaf

Examine a prepared slide of the cross section of a leaf of a representative
eudicotyledon such as lilac (Syringa sp.). Using the lowest magnification on the
microscope, orient the slide so that the section appears as a flat, ribbon-like mass
extending horizontally across your field of view. If the midrib or main vein “bulges”
downward when looking through the ocular, then you have the leaf correctly oriented.

In this section you are seeing all the cell layers that occur from the top to the
bottom of the leaf. The outermost layers at the top and at the bottom are the upper
epidermis and the lower epidermis. All of the cells between them comprise the middle
of the leaf, the mesophyll, that is, the wet culture of chlorenchyma and veins. Change to
higher magnification and examine these in detail.
BIOL 1507E Leaves
The upper epidermis wraps around the leaf and is continuous with the lower
epidermis, but the two differ in certain ways. The upper epidermis is a layer of closely
packed cells. This compact fit helps prevent excessive water loss and provides
mechanical support. The outer walls of the cells are covered by a waxy, noncellular
layer, the cuticle that also retards water loss. The cuticle may be difficult to see in some
slides as the waxy layer can be dissolved during tissue processing. Beneath the upper
epidermis are the cells of the mesophyll containing abundant small bodies, the
chloroplasts. Do the epidermal cells contain chloroplasts? ____

Mesophyll is all the portion of leaf between the upper and lower epidermis. It is
the part that can be likened to a water culture, since it is usually saturated with water and
water vapor. It is composed of chlorenchyma, veins, a certain amount of fibrous tissue,
and a labyrinth of free spaces. Chlorenchyma is a tissue composed of thin-walled
parenchyma cells containing abundant chloroplasts. The colorless walls and glassclear
protoplasm of these cells reflect and refract the incoming light rays, scattering them in
zigzag paths. The ricocheting of each ray lengthens its path and multiplies its chances of
striking a chloroplast before it passes out of the leaf. The chlorenchyma in the upper part
of the mesophyll is known as palisade parenchyma; in the lower part it is known as
spongy parenchyma.

Palisade parenchyma is a layer of evenly spaced, cylindrical cells whose long

axes are at right angle to the upper epidermis. This arrangement puts the cells at an acute
angle to the incoming sunrays that are then not so intense. Full sunlight often inhibits
photosynthesis. So, light intensity that is less than maximal results in more efficient rates
of photosynthesis. Most photosynthesis of the leaf occurs in these palisade cells which
have more chloroplasts than spongy tissue cells. Although they appear to be closely
packed, palisade cells are slightly set apart so that the surface of each is exposed to the
wet atmosphere of the mesophyll. In what practical way does this contribute to the
reaction of photosynthesis?
Usually only one layer of palisade cells is present in the mesomorphic leaf. However,
leaves daily exposed to direct sunlight (as opposed to those partially or periodically
shaded) tend to have two or more layers of palisade. This makes the leaf thicker.

Spongy parenchyma makes up the lower portion of the mesophyll. It is a more

open zone of irregularly shaped cells and large air spaces. Bumps or lumpy extensions of
each cell touch those of other cells. The limited surface contact between cells leaves
much of the cell surface free or exposed. What is the advantage of this?

In growing conditions where the leaf is shaded this random arrangement of cells and their
more or less cube shape favour maximum light absorption. The large air spaces in the
spongy parenchyma allow for rapid, easy flow of carbon dioxide, oxygen, and water
vapour. Specialized pores, called stomata (singular stoma) occur all over the lower
epidermis and control the passage of these gases. There may be the odd stomata on the
upper epidermis.
BIOL 1507E Leaves
Veins are cylindrical strands of vascular tissue and occur mainly in the
mesophyll. They run in all directions, so you will see some in cross-section, circular in
outline and some in profile view (as a more or less complete or incomplete horizontal
band). Examine the large midvein in the center of the leaf. The xylem cells in the vein
are easily recognized by their thick, angular, red-stained walls and absence of cell
contents. Xylem brings water into the mesophyll. Phloem cells are much less distinct;
they are thin-walled, usually of smaller diameter than xylem cells, and located below the
xylem, that is, on the side between the xylem and the lower epidermis.

Study the ring of large, thin-walled parenchyma cells that encircle and may
radiate out from the veins. These cells are the bundle sheath cells. Note that they are
also in close contact with both the palisade and spongy parenchyma. Bundle sheath cells
are important because they are thought to serve as flow channels between veins and
chlorenchyma. This increases conduction in the leaf. Sheath cells envelop the vein over
most of its length and close over and around its tip. In many veins these bundle sheaths
extend to the upper and lower epidermis and help provide mechanical support for the
blade. In large veins sclerenchyma fibers are part of the sheath. If you look at the
midrib you will see above and below it a mass of red, thick-walled sclerenchyma cells.
Such a “reinforced” vein serves as a flexible, yet strong, girder-like support for the leaf.

The lower epidermis is similar to the upper epidermis, except that it has many
small pores or stomata that control the passage of gases in and out of the leaf and the
escape of water vapour. Each stoma is flanked by two sausage-shaped guard cells.
Guard cells are smaller than the adjacent epidermal cells and have a few chloroplasts and,
most noticeably, a very prominent nucleus. Find the guard cells and stomata. Guard
cells and stomata are best seen in an epidermal peel, which you will make later. Changes
in the shape of the guard cells result in an opening and closing of the lens-shaped pore
between them. Such shape changes in the guard cell are due to changes in its water
content. Stomata are opened as a result of water pressure in the guard cells. When light
and heat intensities are high, the leaf can stabilize its temperature somewhat by
evaporating water from its mesophyll out through the stomata.

Prepare a HIGH POWER DRAWING of a Syringa sp. leaf cross section (choose an
area without a vein, only draw the cell outlines, no intracellular detail required and draw
the width of 6-8 epidermal cells wide only!!). Label with the following: upper and
lower epidermis, stoma, guard cell, use a bracket to identify the area of the
spongy parenchyma and palisade parenchyma of the mesophyll. This drawing is to
be handed in for marking prior to leaving the lab. Follow the directions at the end of this
lab manual on how to prepare a high power drawing.

2. Stomata counts
You will observe prepared slides and make epidermal peels from fresh plants to
observe the cells of the epidermis, the guard cells and the variation in the number of
stomata per area between different plants. To prepare an epidermal peel, you will start by
place a drop of tap water on a slide. Using a razor blade, peel a strip of the lower
BIOL 1507E Leaves
epidermis from one of the available plants. Place the epidermal peel with the outer
surface of the epidermis side up in the drop of water on the slide. Place the coverslip on
top. Watch carefully as the teaching assistants will do a demonstration.

Note the large epidermal cells that have different shapes and patterns depending
on the species. In Sedum sp. the cells have wavy contoured cells and are arranged like
pieces of a jigsaw puzzle; in grasses the cells are rectangular and arranged in parallel
rows. Observe the guard cell pairs and the pore between them. Do the guard cells have
chloroplasts? YES. NO.

Different species of plants have different numbers of stomata per unit area of leaf.
Using fresh plant specimens and prepared slides, count the number of stomata seen on the
epidermal peel of three different species. To be able to compare the number of stomata
per unit area you will need to use the same magnification for each specimen you observe.
Therefore you can look at the specimens either at 100X or 400X magnification. Place
your observations in Table 1.

Table 1 Stomata counts

Plant name Magnification # stomata / field

3. The high-efficiency leaf

The monocotyledons most valuable to man are those known as the grasses. Chief
among these are sugar cane and the cereals, corn, wheat, rice, sorghum, etc. It is now
known that many grasses, notably sugar cane, corn, and sorghum, have evolved more
efficient photosynthesis reactions, particularly those trapping carbon dioxide. This
explains their extreme value as a food source. Other plants with these high rates include
some major weeds notably pigweed, lambs-quarters, tumbleweed, and purslane. The
more compact mesophyll of the leaves of these grass crops and weeds maximizes
conduction between the cells and also minimizes any possibilities of drying. Numerous
veins provide each cell with more direct access to water. Big sheath cells around veins
actively accumulate starch in specialized chloroplasts, and their close contact with veins
aids in rapid removal of the photosynthate. All this adds up to higher rates of
photosynthesis which results in greater yield and more vigorous growth. The corn leaf is
a good example of a high efficiency leaf.

Another feature in the corn leaf is the large, bubble-like bulliform cells. These
have thin walls and occur in the epidermis. During the hottest part of the afternoon, or in
BIOL 1507E Leaves
droughts, when water losses from the leaf are high, bulliform cells lose water and turgor
more rapidly than other epidermal cells. As a result, they shrivel causing the leaf to roll
or fold up. This reduces evaporation from those stomata on the leaf surface now enclosed
in the roll. Bulliform cells occur in all grasses and most monocotyledons.
Study a prepared slide of a cross section of corn (Zea mays) leaf. Draw a diagram
of the corn leaf in the following box, showing the special features listed above and
answer the following questions regarding the high efficiency leaf of corn.
1. Is the mesophyll differentiated into palisade and spongy tissue? YES. NO.

2. The vertical position of the corn leaf reduces the need for palisade cells. Explain
how this is so.

3. Are there as many large air spaces in the mesophyll as in the eudicotyledon leaf
previously studied? YES. NO.

4. Are the veins (a) more numerous or (b) less numerous than in the eudicotyledon
leaf previously studied?

5. Would you say that the mesophyll is more compact? YES. NO.

6. Are there bundle sheaths around each vein? YES. NO.

7. Are these sheaths (a) large and prominent or (b) small and inconspicuous?

8. Are the chloroplasts of the sheath cells (a) larger or (b) smaller than those of other
mesophyll cells?

9. Do the bulliform cells occur on one side of the leaf or on both sides of the leaf?
Explain why they would be placed in this fashion?

corn leaf c.s.

B. The Xeromorphic Leaf

BIOL 1507E Leaves
The xeromorphic leaf is one that is designed so that its water losses are
minimized. Plants that grow in dry places or in the actual desert (including the arctic
cold desert) have xeromorphic leaves. Other plants that live in the wet tropics also have
xeromorphic leaves. Surprising? No. In this case it is an adaptation for survival during
the annual dry season.

The special tissue modifications for conserving water in the xeromorphic leaf are:

1. Thick cuticle
2. Epidermal cells with thick walls
3. Several layers of sclerenchyma below the epidermis and in other regions
4. Sunken stomata, that is, stomata inside grooves or cavities recessed down from
the leaf surface
5. Stomata cavities are often lined with hairs
6. In some plants hairs on the leaf surface
7. A more compact and uniform mesophyll with very few air spaces
8. In some plants, palisade tissue on both sides of the leaf

1. The xeromorphic leaf of Marram grass (Ammophila sp.)

Marram grass (Ammophila sp.) is adapted to the dry environment of sand dunes.
The leaf is rolled which makes for a low ratio of exposed surface to volume. This
reduces water loss. Think about it. Which will dry out less quickly: a rolled up wet
towel or wet towel laid out flat?

Examine a slide of a c.s. of Ammophila sp. leaf. Note the inrolling of the leaf.
The xerophytic leaf has furrows on the upper surface in which the stomata occur.
Abundant sclerenchyma is present surrounding the vascular bundles. Note the
compactness of the photosynthetic mesophyll.

Review the list of xeromorphic features listed above and list those present in the Marram
grass:

How does the Marram grass compare with the corn leaf?

Prepare a LOW POWER PLAN of the Ammophila sp leaf cross-section, showing the
arrangement of tissues. Draw 3 of the “bumps” and not the complete leaf. Label with the
following: superior (or upper) epidermis, inferior (or lower) epidermis,
photosynthetic mesophyll, sclerenchyma, stoma, vein. This drawing is to be handed in
for marking prior to leaving the lab.
BIOL 1507E Leaves

C. The Hydromorphic Leaf

The hydromorphic leaf is found in plants that grow either submerged in water or
with their roots continually in wet soil and their leaves floating on the surface of the
water.

Review the list of modifications given for the xeromorphic leaf. Think about the
different conditions impinging on a leaf floating on water and fill in Table 2 with a one-
word or one-phrase description for each of the features that you would expect to find in a
hydromorphic leaf.

Table 2 Modifications Expected to Occur in a Hydromorphic Leaf

Leaf Feature Modification in Hydromorphic Leaves

1. Cuticle
2. Wall of the epidermal cells
3. Position of stomata
4. Amount of air spaces in mesophyll
5. Amount of sclerenchyma
6. Amount of veins (few or many)

Do a quick sketch of the hydromorphic leaf in the box, showing its special
features.
Nymphae sp. hydromorphic leaf