Activity 11

Transport of Water and Other Materials In and Out of the Plants

One of the basic necessities of life is the ability to transport materials that are
needed by the cells to perform their function. Plants accomplish this task through
their vascular tissues namely the xylem for water transport, and phloem for
sugar and other mineral transport. There are two types of transport at cellular
level: passive transport, the movement along a concentration gradient and
active transport, the movement of particles across a concentration gradient
which requires ATP expenditure. Equilibrium must be maintained, thus the cell
membranes play their role of regulating the transport processes undertaken by
the cell.
Objectives:

I.

To determine the mechanisms undertaken by plant to transport the

materials that it needs
To demonstrate the processes by which materials are transported
and transpired
To identify the effects of different factors in the rate of diffusion
To demonstrate how different factors affect the integrity of
membranes
To demonstrate how plants transport water through the stems
To identify different factors that affect the rate of transpiration in
plants
To differentiate guttation from transpiration
Diffusion of Selected Plant Pigments
A. Introduction
Diffusion is a type of passive transport; it is the random movement of
particles in a solution from regions with higher concentration to regions
with lower concentration. Diffusion is very important to plants. Carbon
dioxide required for photosynthesis enters the plant through diffusion in
the in intercellular spaces, oxygen is liberated from plants through
diffusion, and roots absorb water and minerals through diffusion.
B. Methodology
1.
Put 1 g of atsuete seeds each into four test tubes.
2.
Do the ff. in each test tube:
Test tube 1 put 10 ml of distilled water
Test tube 2 put 10 ml distilled water and place in
boiling water bath
Test tube 3 put 10 ml vegetable oil
Test tube 4 put 10 ml heated vegetable oil
3.
After 30 minutes, shake the test tubes and compare
the color intensities with +, ++, +++.
C. Results

Substance
Test tube 1
Test tube 2
Test tube 3
Test tube 4

Color Intensity
+
+++
++
++++

D. Discussion
Questions:
1.
What is the effect of concentration on the rate of
diffusion?
When concentration is higher, the rate of diffusion is
faster. The greater the difference in concentration
between two areas, the greater the rate of diffusion.
2.
What is the effect of size of diffusing particles on the
rate of diffusion?
When particles are larger, there is a slower rate of
diffusion. Consequently, smaller particles diffuse faster.
3.
Which plant pigment traveled the farthest? Explain
your answer.
The pigment in test tube 4 traveled the farthest. When
vegetable oil was heated, temperature increases the
rate of molecular movement (kinetic energy), making
them travel farther and faster.
II.

Osmosis
A. Introduction

Osmosis is the diffusion of water across a selectively permeable

membrane driven by a difference in concentration on the two sides of the
membrane.
B. Methodology
1.
Strip off a thin section of the lower epidermis of the
bangka-bankaan
2.
Prepare a wet mount section.
3.
Examine the cell, which is turgid, under LPO.
4.
Without removing the slide, draw off the water by
using a small piece of tissue.
5.
Replace the water with 5% salt solution.
6.
Examine the plasmolyzed cell.
C. Results

D. Discussion
1.
What is the direction of diffusion of water in a turgid
cell? In a plasmolyzed cell?
When cells(as in the experiment) are placed in water,
water diffuses into the cell causing the protoplast to

III.

swell (turgor pressure) and presses against the cell

wall which also presses with an equal force. When
turgor pressure and wall pressure of the cell are equal,
the cell is in a turgid state. The cell and the solution
are now in equilibrium, no net water movement occurs(
water molecules do move but equally in both direction.
When the cell is placed in salt solution, water diffuses
out of the cell. As it continues to lose water, the
protoplast shrinks and no longer presses the wall. The
cell has been plasmolyzed.
2.
Is the cell wall still intact in a plasmolyzed cell?
The cell wall is still intact, only the protoplasm shrinks.
Factors affecting integrity of cell membrane
A. Introduction

Cell membranes are important to proper cell functioning. It consists of a lipid

bilayer having a dula nature in that they are both hydrophilic (water loving) and
hydrophobic (water-fearing). The inner and outer surfaces are hydrophilic and
interact with aqueous solutions while the core is hydrophobic which does not
permit entry of water and polar molecules.
Because of this dual nature the cell membrane serves many functions : it acts a
selectively permeable membrane that allows entry/exit of materials, and it
houses proteins and carbohydrates necessary for cell to cell and extracellular
communication.
Cell membranes are also sensitive to environmental conditions like temperature ,
pH and organic solvents. Since cells work under narrow range of conditions, we
can hypothesize that higher temperatures and extreme low temperatures will
cause severe damage to cell membranes. The same goes for organic solvents
and pH.
B. Methodology
1.
Make 7 separate sections of peel of red apple.
2.
Place three sections in three test tubes (A, B,C) filled
with 10 ml distilled water. Put tube A under room
temperature, B inside a refrigerator, anc C in a water bath
with temperature 60oC.
3.
After 30 minutes, remove the sections and prepare a
wet mount. Examine under microscope and compare color
intensities.
4.
Place the other sections in four slides (D, E,F,G). Add
50% choloroform to D, 50% acetone to E, 0.1M NaOH to F,
and 0.1M HCl to G.
5.
Examine the cells immediately, after 15 minutes and
after 30 minutes.
C. Results
Test tube
A(room temp.)

Color intensity
+++

Test tube
D(chloroform)

Color intensity
++

B(refrigerator)
C(60o C)

++
+

E(acetone)
F(NaOH)
G(HCl)

+
++
+

D. Discussion
1.
Where does the pigment come from? What do you call
this pigment?
These pigments are called anthocyanins. They are
stored in vacuoles.
2.
In order to reach the outside of a cell, through what
membranes must the pigment diffuse?
The anthocyanin pigment must diffuse out of the
tonoplast (or the vacuolar membrane) and the cell
membrane itself.
EFFECTS OF TEMPERATURE
3.
What temperature stressed and damaged the
membranes the most? Explain.
The highest temperature (60o C) stressed and
damaged the membranes the most as shown by the
great change in color of specimen after being heated.
Specimen A has shown the greatest color intensity, its
color close to the original, showing that in room
temperature there is little to no disruption of
membrane. B follows which shows that when subject to
cold temperature, cell membrane are disrupted but not
as much as when exposed to heat.
When you apply heat, you apply energy which makes
molecules vibrate rapidly. Lipids become more fluid,
proteins untangle and the membrane breaks apart. The
cell membrane is damaged and pigments will leak out
of their compartments which explains the apparent
loss of color.
EFFECTS OF ORGANIC SOLVENT
4.

Which solvent stressed the membranes more? Explain.

The color intensity of E which is treated with acetone
is lighter than D which is treated with chloroform. This
means that acetone is the stronger organic solvent and
has done more damage to the cell membrane.

EFFECTS OF pH
Because cells are usually exposed in normal pH levels,
exposure to very basic pH levels (from NaOH) and very acidic
pH levels (from HCl) causes cell membrane to break and
pigments to leak out.

IV.

Imbibition
A. Introduction
Imbibition is the disruption of water accompanied by swelling.
B. Methodology
1.
Individually weigh 2 pieces of wood and rubber, and 2
sets of 10g of corn seeds.
2.
In the first beaker, immerse one piece of wood,
rubber, and 10g of corn seed in water. In the second beaker
immerse one piece of wood, rubber, and 10g of corn seed in
kerosene.
3.
After 90 minutes, take out the materials and get their
final weight.
C. Results
Medium

Imbibitant

Water
Kerosen
e

Rubber
Wood
Seeds
Rubber

Initial
weight
0.5g
0.8g
10g
0.5g

Final
Weight
0.5g
1g
10.35g
0.8g

% of change in
weight
0%
25%
3.5%
60%

Wood
seeds

0.8g
10g

1.25g
10.75g

56.25%
7.5%

D. Discussion
1.
Which of the materials tested serve as good imbibitant
of water? Kerosene?
Both wood and seeds imbibed water but wood is the
better imbibitant. All materials absorbed kerosene but
wood and rubber are better imbibitants of water.
2.
Describe the nature of the chemical constituents of
water? Kerosene?
Water (H2O) is a polar substance (with partial positive
and partial negative charge) while kerosene is anonpolar.
3.
Are living cells necessary in order for imbibitions to
take place?
No. wood and rubber imbibed kerosene.
4.
Are there any living cells in the veneer?
Yes.
5.
In the seeds, are living cells involved in imbibitions?
Are dead cells involved?
The seed is composed of living cells which undergo
imbibitions. Dead cells are also involved; the see
coat/testa is composed of dead cells which imbibe
water and break apart.

V.

6.
In what way/s is the swelling effect of imibition
important to seed germination?
Imbibition is the initial step in seed germination. It
causes the seed to swell and results in breaking of the
testa.
Movement of water through the stem
A. Introduction
As water is absorbed by roots, it moves through the stem through the
xylem. The xylem transports water to the whole of the plant body. This
experiment will trace the movement of water through the xylem in the
plant.
B. Methodology
1.
Prepare pechay leaves with petiole. Cut off 1 cm from
the base of the petiole.
2.
Immerse the petioles in a beaker containing 10 ml of
1% eosin dye solution.
3.
After 10-15 minutes, remove the leaf and split the stalk
longitudinally to measure the length covered by the dye.
4.
From another leaf, obtain a cross section of the petiole
and examine under the microscope.
C. Results

<PICTURE NUNG X-SECTION NG PECHAY STALK>

D. Discussion
After 15 minutes, the stem all the way to the veins of the pechay leaves
have red stain indicating that the eosin dye solution where the stem is
immersed have been absorbed and transported by the xylem( as seen in
x-section, vascular bundles have bright red stain).

VI.

Comparison of cuticular and stomatal transpiration by four leaves method

A. Introduction

Transpiration is the evaporation of water through the stomata of the plant. The
stomatal frequency and size is directly proportional with rate of transpiration
the more stomata and the bigger their size, the faster the rate of transpiration.
Also, the stomata in the abaxial surface contain more stomata which helps in
conservation of water. Stomata in the adaxial surface are considerably fewer. By
applying petroleum jelly on the different surfaces of the four identical leaves, we
can test the resulting dryness of the leaves.
B. Methodology
1.
Prepare four identical leaves and label them A, B, C, D.
2.
Leaf A is the control. On leaf B, apply petroleum jelly
on the upper surface. On leaf C, apply petroleum jelly on the
lower surface. On leaf D, apply petroleum jelly on both sides.

3.
Hang the leaves on a thread where both surfaces are
exposed to air. Keep the set-up until the next lab period.
C. Results
After a few days upon exposing the four identical leaves in the air, specimen A is
the driest, followed by B, C, and D is the least dry.
D. Discussion
Specimen A became the driest because it had no petroleum jelly to cover its
stomata. Rate of transpiration remained the same. Leaf B is less dry than A
because petroleum jelly covered some stomata on the adaxial side but not the
abaxial side which contained more stomata. This decreased the rate of
transpiration. Leaf C is less dry than B because petroleum jelly is applied in the
abaxial side which covered more stoata, thus reducing even more the rate of
transpiration. Leaf D is the least dry because the petroleum jelly covered both
abaxial and adaxial sides, reducing the transpiration the most.
VII.

Guttation
A. Introduction

Guttattion is the exudation of water drops through the hydathodes. At night,

stomata are closed and there is a high soil moisture. Water will enter the plant
roots which will create a slight root pressure. This pressure forces some water to
exude through the hydathodes in the leaf margin.
B. Methodology
1.
Plant 5 viable grains in a flower pot with holes at the
bottom.
2.
Immerse the lower portion of the tin can in a shallow
dish. Add water from time to time.
3.
When the seedling are 2-5 cm long, cover the flower
pot including the dish with a transparent bell jar. Take note of
the droplets formed on the leaf surfaces.
C. Results

Guttation is observed in the leaf margin.

D. Discussion
1.
Explain how guttation was induced in the rice
seedlings.
Adding water from time to time creates a high degree
of moisture wherein water enters the roots of the rice
seedlings. This will create a slight root pressure. Root
pressure forces the water to exude in the leaf margin.
2.
Differentiate guttation from transpiration.
In both guttation and transpiration, plant loses water.
Guttation is induced by the slight root pressure
because of high soil moisture while transpiration is
caused by heat in the plants environment. In
guttation, water escapes through the hydathodes and
tends to accumulate in droplets in the leaf margin
while in transpiration, water escapes through the
stomata and evaporates.

References:
Botany by Mauseth
http://plantcellbiology.masters.grkraj.org/html/Plant_Cellular_Physiology1Water_And_Water_Dynamics.htm
http://answers.yahoo.com/question/index?qid=20061129041445AAaMGDT
http://www.tutorvista.com/content/biology/biology-iv/plant-waterrelations/imbibition.php#

http://en.wikipedia.org/wiki/Guttation
http://lariceman.wordpress.com/tag/guttation/

Activity 12
Introduction:
Photosynthesis is a complex process by which carbon dioxide is combined with
water to form carbohydrate. It converts light energy into chemical energy. And
during the process, liberation of oxygen occurs. Chlorophyll, an essential
pigment, allows photosynthesis to take place.

Objectives:
At the end of the exercise, the student should be able to:
1. To know the importance of chlorophyll, light, and carbon dioxide in
photosynthesis
2. To observe the release of oxygen in photosynthesis
3. To perform separation of chloroplast pigments; to identify these pigments
I. Hypotheses:
A. If the leaf is positive for starch test (IKI solution turns blue-black) then it is
photosynthetic.
B. If the part of the leaf which was pre-covered with black paper is not
positive to starch test then that particular part has not been
photosynthetic.
C. If the color intensities differ then the CO2 concentration in each of the test
tubes set up also differ.

D. If the lighted match continues to burn when placed in the test tube then
oxygen is present.
E. If the lengths travelled by the pigments differ then they have different
rates of solubility.
II. Methodology:

A. Boil variegated leaf in water. Immerse the leaf in a test tube with 95%
ethyl alcohol. Place the test tube in hot water until pigments are
extracted. Then wash the leaf with water and using IKI solution, test for
the presence of starch.
B. Place a potted plant in the dark for 48 hours. Then select a few leaves and
wrap portions of the leaves with black carbon paper. Expose the whole
plant to light for 5 hours. Detach the covered leaves and test for the
presence of starch.
C. Fill test tubes A, B, & C with previously boiled then cooled water. Place
Hydrilla sprigs in the tubes A & B then add a pinch of NaHCO3 in B & C.
Expose A, B, & C to bright light. Observe if bubble formation occurs.
Remove Hydrilla from A & B. Add five drops of phenol red to each test
tube. Then compare color intensities.
D. Fill a beaker with of a liter of water. Using plastic straw, blow air gently
into the beaker. Cut the ends of Hydrilla under water and insert thistle
funnel over the sprigs (cut ends should face the tube of the funnel). Place
a test tube filled with water upside down over the tube of the funnel
partially dipped in the water of the beaker. Observe for bubbles emerging
out from the cut ends. Determine the nature of the gas by placing a
lighted match in the test tube.
E. Prepare the leaf extract of mature Hibiscus leaves. Prepare the filter paper
strip. Prepare the chromatogram. Place the paper strip on the hook and
insert into the test tube containing solvent mixture of 95 parts petroleum
ether and 5 parts acetone (solvent level must be below the 1cm mark).
Replace cork with the strip and observe flow of solvent until the 2cm mark.
Remove the paper from the tube and let it dry.

III. Results:
A. Upon dropping IKI solution onto the bleached leaf, blue-black color
developed. This denotes the presence of starch which is actually an
indicator that the leaf is photosynthetic.
B. Covered parts of the leaves were negative for starch test while the rest of
the parts were positive.
C. Color intensities:
Test tube A (with Hydrilla but no sodium bicarbonate) - remained red, no
bubbles formed
Test tube B (with Hydrilla and sodium bicarbonate) - became yellow but
returned to red, bubbles were formed

Test tube C (NaHCO3 only) turned from red to yellow.

D. Bubbles emerged from the cut end of the Hydrilla. Then when the lighted
match was placed near the mouth of the test tube (immediately after it
was removed from the water) the flame on the match kept on burning.
E. After the experiment, four pigments were visible: yellow orange
(carotene), light yellow (xantophyll), blue green (chlorophyll a) and olive
green (chlorophyll b). The chromatogram also showed that carotene
travelled the farthest, followed by xantophyll, chlorophyll a and finally
chlorophyll b.

IV. Discussion:
A. Starch is a product synthesized from chlorophyll and is therefore found in
green (photosynthetic) parts of the leaf. Boiling the leaf prior to chlorophyll
extraction removes the cuticle (which may prevent the entry of IKI
solution), damages cell membranes to make starch (in the cytoplasm &
chloroplast) accessible to IKI solution, and arrests all chemical reactions.
Boiling a leaf in alcohol removes its chlorophyll, so the leaf loses its
green color. A leaf positive to starch test indicates that the leaf is
photosynthetic obviously because sugar is a product of photosynthesis
which is stored as starch in almost all parts of the plant.
B. Light is the key to jumpstart photosynthesis. It provides the energy
needed to perform light reactions. Sample A (leaf grown under normal
conditions) contained more starch than sample B (leaf with covered
portions). First is because Sample B was kept in the dark for 48 hours to
make it consume its reserved starch. Second reason is that covered
portions were not permitted to perform photosynthesis due to the absence
of light. Unlike the exposed parts of the leaf, covered ones were really
expected to be more or less negative to starch test.
C. NaHCO3 (Sodium bicarbonate) provided a good source of CO2 for the
Hydrilla in test tube B maximizing its photosynthetic ability. This lowered
the pH of the water and caused phenol red to turn into yellow (since
Phenol red is an indicator of p H). Indirectly, it is an indicator of CO2
concentration because CO2 in water forms carbonic acid which lowers
the pH of water. However, the Hydrilla used up the CO2 in the solution and
gave off oxygen gas (indicated by the bubbles) which caused the solution
to become neutral again (red). Set up A also actually exhibited the use of
CO2 in photosynthesis, but since there was no source present (NaHCO3)
the reaction was limited. In set-up C (Na 2HCO3 only), the solution became

acidic and it turned yellow because of the presence of sodium bicarbonate,

which dissociated to form CO 2 and ultimately, carbonic acid without any
plant to use the CO2 up and produce oxygen. Boiling dissolved the gases
present in the water.
D. Oxygen is a by-product of photosynthesis which ultimately, comes from
the water absorbed by the plant. When the test tube was lifted and a
lighted match was placed near its mouth, the match continued to burn.
This just indicates the presence of oxygen (because combustion needs
oxygen). And since the life the flame on the lighted match was sustained,
it showed that oxygen is present in the gas (or bubbles) produced by the
Hydrilla sprig.
E. Chloroplast pigments are known as chlorophylls (green pigments) and
carotenoids (yellow-orange-red pigments) . Different pigments absorb
different light wavelengths. Solubility of the pigments varies according to
the rate they are carried by the solvent. The colors visible in the
chromatogram are dark green (chlorophyll a), light green (chlorophyll b),
and yellow (xantophyll). Chlorophyll b is the least soluble and xantophyll is
the most soluble. The solvent (petroleum ether with 5 parts acetone)
carries the pigments as it moves up the paper. The pigments are carried
along at different ratesdue to their differences in solubility, the less soluble
pigments will move slower up the paper than the more soluble pigments.
Each pigment has an Rf value, the speed at which it moves over the paper
compared with the speed of the solvent.
Rf = Distance moved by the pigment
Distance moved by the solvent
Distance of entire solvent = 6.7cm
Rf value of chlorophyll a= (6cm/6.7cm) = 0.89
Rf value of chlorophyll b = (5cm/6.7cm) = 0.75
Rf value of xantophyll = (6.7cm/6.7cm) = 1
References:
http://www.nuffieldfoundation.org/practical-biology/testing-leaves-starchtechnique
http://www.brilliantbiologystudent.com/testing_a_leaf_for_starch.html

http://www.markedbyteachers.com/gcse/science/testing-starch-in-a-variegatedleaf-lab-report.html
http://breakthelight.wordpress.com/tag/test-tube/
An Introduction to Plant Biology (Fourth Edition) by James Mauseth