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COLLEGE OF SCIENCE

DEPARTMENT OF BIOLOGICAL SCIENCES

Experiment 1

CELL WATER POTENTIAL

The tendency of water to move into or within a system, such as a plant or animal tissue,
the soil or the atmosphere can be measured as the amount of energy per unit volume (or
pressure). Such values, expressed in units of bars or megapascals (MPa) is referred to as the
water potential of the cell and may be computed mathematically as:

Equation (1 –1): Ψ = Ψs + Ψ p + Ψ m

where: Ψ = water potential of a cell; Ψ = 0 at equilibrium;


Ψs = solute or osmotic potential; always negative in sign;
Ψp = pressure potential; turgor pressure; may be (-), 0, or (+);
Ψm = matric potential; due to water-binding colloids in the cell; negative in sign.
Plant cells usually have a zero (0) water potential when they are in equilibrium with pure
water (turgid). Dissolved solutes contribute to the water potential.
The objective of this experiment is to measure the water potential and osmotic potential
in a tissue, then calculate the pressure potential. For practical reasons, a plant tissue, instead
of animal tissue, is used in this experiment.

Materials:
Plant samples (potato, sweet potato, radish, turnips, cucumber)*
Sugar solutions (0.1m; 0.2m; 0.3m; 0.4m; 0.5m; 0.6m; 0.7m)
Distilled water
Balances and weighing paper
Plastic cups
Basin
Sugar
Salt
Funnel, 150mm
Erlenmeyer flasks, 250 ml
Paper towels/filter paper
Blender
Heidenhain (freezing) thermometer / Radioshack Infrared Thermometer
Chopping board
Magnetic stirrers with stir bars
Parafilm
Knife
Ruler
Cheesecloth

*Laboratory instructor may modify this list. Do not wash the plant samples.

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COLLEGE OF SCIENCE
DEPARTMENT OF BIOLOGICAL SCIENCES

PART I. DETERMINATION OF WATER POTENTIAL, Ψ


When a tissue is placed in a sugar solution, there will be a net movement of water into or
out of the tissue depending on the relative water potentials: into the tissue with a hypotonic
solution; out of the tissue in a hypertonic solution; or no net movement in an isotonic solution
when the tissue and solution are at equilibrium. If a series of solutions of different
concentrations of sucrose is used, the sucrose concentration that would cause no change in
weight may be determined by plotting the percent change in weight against sucrose
concentration. Such solution may be assumed to have a water potential equal to that of the
tissue.
Procedure:
1. Prepare eight (8) different sucrose concentrations (dH2O, 0.1m, 0.2m, 0.3m, 0.4m, 0.5m,
0.6m, 0.7m) and dispense 75ml of each separately into 150ml beakers / plastic cups.
2. Cut plant samples into small pieces approximately 1 cubic centimeters each. Place 3 – 4
cubes in covered beakers to prevent them from drying out. Save remaining plant samples
for Part II.
3. Blot cylinders with paper towels and weigh (to 0.001g) in sets of 3 - 4. Record the weight.
4. Put one set of cubes in each of the beakers with sucrose solutions prepared in Step 1.
5. After 45 minutes to 1.0hr, remove the cubes, blot with paper towels and weigh again.
Record results as before.
6. Compute for the change in weight (∆W) and the percent change in weight (%∆W) using
the following formulae.

Equation (1–2): ∆W = W f − Wi

∆W
Equation (1–3): % ∆W = x100
Wi
where: Wi = initial weight of the cylinders;
Wf = final weight of the cylinders.
7. Tabulate all results. Plot % change in weight vs. sucrose concentration. Draw the best-fit
straight line through the points.
8. Determine molal concentration of sucrose that gives 0% change in weight. Compute for
Ψs in bars of that sucrose solution using the formula:
Equation (1–4): Ψs = -miRT
where: m = molality; (NOTE: 1 molal = 1 x 103 mol m-3 H2O)
i = ionization constant = 1 for sucrose
R = gas constant = 8.31 J K-1 mol-1
T = room temperature in K (oC + 273)
NOTE: For uniformity, room temperature is computed at 26oC

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COLLEGE OF SCIENCE
DEPARTMENT OF BIOLOGICAL SCIENCES

9. Determine the water potential of the potato assuming that Ψm is very small and therefore
negligible. Convert unit of J m-3 (energy per unit volume, pressure which is equivalent
to Pa) to MPa by dividing computed value by 106.
NOTE: In a free standing solution, Ψp = 0; so Ψ = Ψs.

PART II. DETERMINATION OF SOLUTE POTENTIAL (ΨS) OF EXTRACTED SAP BY CRYOSCOPY


Cryoscopy (cry·os·co·py) is the examination of liquids, based on the principle that the
freezing point of solutions varies according to the amount and the nature of the solute. This
provides a relatively easy means of arriving at the osmotic potential of a solution. The
freezing point of an aqueous solution decreases with an increase in dissolved solutes. Once
the freezing point of a solution has been determined, its solute potential may be calculated,
thus:

Equation (1–5): Ψs = 1.22Tƒ


where: Ψs = solute potential in MPa at 0oC
Tƒ = freezing point of the solution in oC
The equation holds true for freezing temperature. The solute potential of the extracted sap
at room temperature can then be computed:
Equation (1–6): Ψs = 1.22Tƒc
where: c is the correction factor; (Rm temp in K/273K)

Procedure:
Sap extraction
1. Peel 2 – 4 potatoes, chop and place in a blender. Leftovers from Part I may be used. Puree
the tissue.
2. Filter blended potato with cheesecloth to remove cell wall and debris. Store in covered
beaker / plastic cup.
Freezing point determination
3. Immerse Heidenhain thermometer in crushed ice – salt bath. Record reading.
4. Pour 60ml sap in 250ml Erlenmeyer flask with magnetic stirring bar and insert
thermometer. Surround flask with ice–salt bath (see Figure 1–1). Stir vigorously.
5. When temperature reads about 1oC, read temperature every 10 sec.

NOTE: There should be a continuous drop in temperature, followed by a short plateau


which corresponds to the freezing point, is followed by an increase.

6. Plot temperature vs. time.

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DEPARTMENT OF BIOLOGICAL SCIENCES

Heidenhain thermometer

Erlenmeyer flask Rubber stopper

Beaker

Ice-salt bath
Stir bar
Sap extract
Magnetic stirrer 4
5 6
7 4
5 6
7

3 8 3 8

2 9 2 9
1 11 1 10

Figure 1 – 1. Setup for freezing point determination

7. Determine the true freezing point, Tƒ.


Equation (1–7): Tƒ = Tƒ’ – 0.0125ts
where: Tƒ’ = apparent freezing point
ts = degrees of supercooling; (-) in sign
ts = lowest temp - Tƒ’
0.0125 = amount of water (1/80) solidifying per degree of
supercooling

8. Make another correction for the zero point of the thermometer.


9. Determine the solute potential using Equation (1–5) at 0oC.
10. Calculate the pressure potential (Ψp) of the cells of plant sample.

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COLLEGE OF SCIENCE
DEPARTMENT OF BIOLOGICAL SCIENCES

Group No. ____ Date Performed: ____________________


Group Members: Lab Instructor(s):____________________
___________________________ ______________________
___________________________ Signature(s): ______________________
___________________________
___________________________
___________________________

Experiment 1

CELL WATER POTENTIAL

I. Determination of Water Potential


Change in % Change
Sucrose Initial Weight Final Weight
Weight in Weight
Concentration In Grams (Wi) In Grams (Wf)
(∆W = Wf - Wi) (%ΔW)
Dist. H2O
0.1 m
0.2 m
0.3 m
0.4 m
0.5 m
0.6 m
0.7 m
Table 1-1. Results for determination of water potential.

Molality of sucrose where ΔW = 0

Show your computations below:

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COLLEGE OF SCIENCE
DEPARTMENT OF BIOLOGICAL SCIENCES

II. Determination of Solute Potential of extracted sap through cryoscopy

Temperature Time Temperature Time


(Celsius) (seconds) (Celsius) (seconds)

Table 1-2. Results for determination of solute potential.

III. Compute for the water potential of your plant sample.

Solute Potential (Ψs) Pressure Potential (Ψp) Water Potential (Ψ)

IV. Summary of Results

Group No. Plant Sample (Common Name, Scientific Name) Ψ


1
2
3
4
5
6
7
8
9
10
Note: Water potential should be indicated up to three (3) decimal places.

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COLLEGE OF SCIENCE
DEPARTMENT OF BIOLOGICAL SCIENCES

Guide Questions:

1. Why is the osmotic potential always negative in sign?


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2. How do dissolved solutes contribute to the water potential of the cell?
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3. What is the sign of the pressure potential of the tissues at equilibrium? (+), (0), or (-)?
At incipient plasmolysis? During transpiration?
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4. What causes supercooling? Why is the lowest point reached not considered the freezing
point of the sap solution?
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5. Do plants with watery sap have higher or lower water potential? Explain your answer
using actual computed values.
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