EXERCISE # 1

PLANT CYTOLOGY
INTRODUCTION:
Plant cells, like animal cells are complex structures with membrane-bound organelles
and a true nucleus. These organelles serve as the control center, powerhouse of the cell,
packaging and transport system, communication system and waste-removal system.
This exercise tackles the basic structures of the plant cell that will be very significant for
the understanding of the plant function.
OBJECTIVES:
At the end of this activity, the students should be able to:
a. relate the different organelles of the cell with its functions
b. understand the effect of different stains on the cell structure
c. differentiate various types of plastids
MATERIALS
Apparatus/Glasswares
Microscope
Beakers with dropper
Slides and cover slips
Scalpel/blade

METHODOLOGY

Reagents/chemicals
Distilled water
5% ethanol
9 N HCl
Saturated solution of
NaCl
Iodine

Specimen/Samples
Twig
Moss
Rhoeo discolor
Tomato/red pepper
potato

Cell wall
1. Make free-hand sections of a twig using sharp razor blade.
2. Observe in wet mount with water first and then mount in one drop of 5% (EtOH)
plus one drop of 9N HCl.
3. Repeat with bean stem to decide which kinds of tissues and cells have walls
containing lignin in plants.
4. Illustrate
Cell Membrane
1. Make a wet mount of a Mnium (moss) leaf and observe the cells in distilled
water.
2. A saturated solution of NaCl is available for a second mount or to add to the first
one. The distilled water will obviously be hypotonic, the salt solution will be
hypertonic.
3. Illustrate
Chloroplasts
1. Make another wet mount of the Mnium (moss) leaf.
2. Add a drop of iodine stain to the mount. A positive reaction with the presence of
of starch is production of blue-black speck.
3. Take note of the number of chloroplasts per cell.
Vacuole
1. Obtain Rhoeo discolor leaves.
2. Peel the upper and lower epidermis. The upper epidermis of the leaf is
essentially transparent but the lower epidermis is quite purple.
3. Make a wet mount of the upper and lower epidermal layers peeled from this leaf
and compare them.
4. Locate the purple pigment.
5. Draw the structures.
Nucleus
1. Continue the observation of the upper and lower epidermis.
2. Using the iris adjustment, find the nucleus of many cell types.
3. Count the nucleus per cell.
4. Draw the cell with the presence of a nucleus.
Proplastids
1. Find another member of the plastid family in association with the nucleus.
Chromoplasts
1. Make free hand sections of red pepper or tomato and make a wet mount.
2. Draw the structures.
Elaioplasts
1. Make wet mount out of potato to reveal elaoiplasts.
2. Draw the structures.
Amyloplasts
1. Make wet mount out of potato.
2. Draw the structures.

RESULTS and DISCUSSION

Cell Wall

Cell wall is the outermost layer of the plant cell. It is a rigid structure which gives
support, protection and shape to the cell. It consists of three layers primary cell wall,
secondary cell wall and tertiary cell wall.

The wet mount of twig with ethanol plus hydrochloric acid (Fig. 1) showed the
rigid structure of the cell wall. 5% (EtOH) with 9N HCl is a stain for lignin, a hard
substance in which the cellulose fibers become embedded. It gives wood many if its
distinctive properties. Therefore, the twig has walls containing lignin. Of all polymers,
lignin is the only one that is not composed of carbohydrate monomers. Instead, its
primary component is coniferyl alcohol.

Cell Membrane

Cell or plasma membrane is another part of the boundary of the cell that confines
the contents of the cell to an internal compartment. It regulates the flow of materials into
and out of the cell with its semi-permeability. It is made up of phospholipid bilayers with
integral and peripheral proteins.

Since plant cells have a cell wall, the membrane is not visible unless it is pulled
away from the wall slightly. Even then, the membrane will not be seen, only its effects in
containing the cytoplasm.

The first mount is with distilled water (Fig. 4) which is hypotonic. When a plant
cell is placed in a hypotonic solution, it takes up water by osmosis and starts to swell, but
the cell wall prevents it from bursting. The plant cell is said to have become "turgid. The
pressure inside the cell rises until this internal pressure is equal to the pressure outside.
This liquid or hydrostatic pressure called the turgor pressure prevents further net intake
of water. Turgidity is very important to plants as it helps in the maintenance of rigidity and
stability of plant tissue.

When a plant cell is placed in a hypertonic solution, like salt solution, the water
from inside the cells cytoplasm diffuses out and the plant cell is said to have become
"flaccid". (Fig. 3) it will be noticed that the cytoplasm has shrunk and pulled away from
the cell wall .This phenomenon is called plasmolysis. The process is reversed as soon
as the cells are transferred into a hypotonic solution (deplasmolysis).

Chloroplasts

Among the several types of plastids, chloroplasts are the most common. These
have a photosynthetic function: converting light energy to chemical energy. Chloroplasts
contain enzymes necessary for photosynthesis and also the green pigment called
chlorophyll, a molecule with the vital role of absorbing light energy.

The mount added with iodine produced a blue-black speck in the micrograph.
(Fig. 2) This indicates a positive reaction with the presence of starch. Approximately 11
chloroplasts are found in each cell.

Vacuole

The upper and lower epidermis of Rhoeo discolor is compared and it is seen that
the upper (Fig. 5) is essentially transparent while the lower (Fig. 6) is quite purple. Stoma
is visible in the lower epidermis along with its guard cells. Along with these, a purple
pigment called anthocyanin is also seen. This water-soluble pigment is stored in the
vacuole of Rhoeo discolor, giving it its characteristic color.

Vacuole (Fig. 7) is a membrane-bounded sac filled with a liquid that contains a

variety of materials in addition to water like dissolved salts, ions, pigments and waste
products. It is most common in plant cells and cells of certain protists.

Nucleus

With some adjustments, the nucleus became visible in the cell of Rhoeo discolor.
(Fig. 8) Nucleus houses the DNA and serves as the control center of the cell.

Proplastids

Proplastids (Fig. 9) are the colorless precursors where the other plastids have
developed. These have the ability to divide and differentiate to various types of plastids.

Cells that lack chloroplasts can still obtain chemical energy by the aid of the other
members of the plastid family since they can change from one type to another,
depending of the needs of the plant cell.

Chromoplasts

In the wet-mount of tomato (Fig.10), there are colored pigments seen and these
are the chromoplasts. They synthesize food materials by photosynthesis. Tomatoes and
red pepper turn red on ripening due to the formation of the red pigment called lycopin
replacing the chlorophyll.

Elaioplasts

The wet mount of potato (Fig. 11) showed non-pigmented plastids called
elaioplasts, a type of leucoplast that is specialized in storing lipids and housing body oil
deposits as rounded plastoglobuli.

Amyloplasts

Another type of leucoplast is found in the free-hand section of potato (Fig. 12).
These are specialized for the storage of starch and found mostly on tubers, rice, and
wheat.

REFERENCES

Reece, Jane B. et. al. Campbell Biology 9th ed. Pearson Education, Inc. 1301 Sansome
St., San
Francisco, CA 94111. 2011

Berg, Linda R. Introduction to Botany Philippine Edition. Cengage Learning Asia Pte Ltd.
Pasig City, Philippines 1605. 2014