Experiment 1:

Cell Water Potential

Christine Marie H. Ulbis, Adrionna Faye Pauline A. Uy, Gian L. Veloso, Bianca T. Villagomez

Department of Biological Sciences

College of Science, University of Santo Tomas
Espaa, Manila Philippines

Abstract
The water potential was measured for each root crops. It resulted to values, -0.7220, -0.7059, -
1.0999, -0.8191, -1.2148, -1.2271, -1.3608, -1.3779, -0.6693, -1.0121 from groups 1-10, respectively.
Similarly, the osmotic potential for each root crop was also measured, the values for camote ranges from -
0.6547 MPa to 3.4722 MPa, for potato it ranges from 0.1325 MPa to 0.4695 MPa, for carrot it ranges
from 1.1002 MPa to 1.7801 MPa, for Jicama it ranges from 2.0447 MPa to 3.8304 MPa and for Radish it
ranges from 0.4294 MPa to 1.0089 MPa..

Keywords: Osmotic Potential, Water Potential, Pressure Potential, Freezing Point, Root Crops

Introduction
Cell Water Potential, symbolized by (the
Greek letter psi), is defined as the energy required
for water to move from the sample and into a
relatively pure free water in reference conditions.
Through the process of osmosis, water moves
from a region of high area of concentration to a
region of low area of concentration. Additionally,
concentration, pressure and gravity are major
contributing factors in a calculating a plants
water potential (Sinauer, 2015).
Cryoscopy is the determination of the
depression in freezing point unto which
molecular weights of the dissolved solutes can be
determined. The freezing point occurs when the
temperature of the liquid state of a liquid
substance is in equilibrium to its solid state. It has
a colligative property meaning the freezing point
lowers when a compound or more dissolved
solutes are added. Once the freezing point is
determined, it can be used to calculate for the
solute potential which is part of the equation in
calculating for the plants water potential.
This experiment aims to measure the water
potential and osmotic potential in a plant tissue
and then calculate the pressure potential.
Methodology
Eight different sucrose concentrations
(dH2O, 01.m, 0.2m, 0.3m, 0.4m, 0.5m, 0.6m,
0.7m) were prepared. 75mL of each were
dispensed separately into plastic cups. Sixteen
cylinders were bore from the root crops (different
plants were assigned on each group) using a No.
5 cork borer. Cylinders were placed in covered
plastic cups to prevent them from drying out.
Each cylinder was cut approximately 4cm and the
remaining plant samples were saved for Part B of
the experiment. The cylinders were blotted with
paper towels and weighed in sets of two. The
weights were recorded. One set of cylinders was
put in each of the beakers with the said sucrose
concentration solutions. After 1.5 hours, the
cylinders were removed, blotted with paper
towels and weighed again. The weights were
recorded. The change in weight and the percent
change in weight were then computed. The
results were tabulated and the percent change in
weight versus sucrose concentration was plotted
with the best-fit straight line drawn through the
points. The molal concentration of sucrose that
gives 0% change in weight was then determined.
The water potential of the root crops were the
determined.
 The remaining root crops from Part A of
the experiment were placed in a blender. The
tissue was pureed. The blended root crops were
filtered with cheesecloth to remove cell wall and
debris, and was then covered in beaker. 60mL sap
was poured in 250mL Erlenmeyer flask with
magnetic stirring bar and the thermometer was
inserted. The flask was surrounded with ice-salt
bath and was stirred vigorously. After the
temperature was 1oC, the temperature was
recorded every 10 seconds. The data was
recorded and the temperature versus time graph
was constructed. The true freezing point and
solute potential was then determined. The
pressure potential of the cells of the root crops
were then calculated.
Based on the computed values for Parts A
and B, the water potential of the root crops were
determined. The values for all plant samples were
then compared.
Results and Discussion
In theory, no net gain or loss of water in
tissue after immersion in a solution of known
molarity means that its water potential is equal to
that of the external solution.
Employing the Gravimetric Technique,
water potential values may be negative due to the
infiltration of water and solutes within the
apoplast (Saupe, 2009). Bland and Tanner (1985)
reported that the osmotic potential for a potato
tube is in the range of -0.58 to -1.53 MPa. The
root crops should fall within that range.
In this experiment, the samples were
immersed in solution with a variation of sucrose
concentration resulting to the losing or gaining of
water by the process of osmosis. Osmosis is the
spontaneous net movement of solvent molecules
through a semi-permeable membrane into a
region of higher solute concentration (Haynie,
2001). If the samples were placed in a solution of
higher solute concentration, (hypertonic solution)
the water will go out of cell, thus the decrease in
the weight of the samples. In contrast, if the
samples were placed in a solution of lower solute
concentration, (hypotonic solution) the water will
go inside the cell, thus the increase in the weight
of the samples. This is because dissolved solutes
contribute to the osmotic pressure or potential.
Solutes reduce water potential by consuming
some of the potential energy available in the
water.
According to Kramer & Myers (2012),
osmotic pressure is the external pressure required
to be applied so that there is no net movement of
solvent across the membrane (isotonic). It is a
colligative property which depends upon the
concentration of solute molecules or ions, but not
upon the identity of the solute. In these
calculations, molality (m) is used which is
defined as the number of moles dissolved per
kilogram of solvent. This is because the mass of
solvent doesn't change with temperature unlike
the volume. This is the reason why molality is
preferred over molarity (moles per liter).
Osmotic pressure and potential, in turn,
affects water potential. Water potential is the
tendency of the water to move into or within a
system measured as the amount of energy per unit
volume expressed in megapascals (MPa). Water
potential can be computed mathematically as:
= + +
where is the water potential of a cell, is the
solute or osmotic potential, is the pressure
potential and is the matric potential.
The pressure potential (p) is the effect
of hydrostatic pressure on the potential energy of
a solution. It is defined as 0 MPa for STP
(absolute pressure of 1 atm = 0.1 MPa). For a case
of a partial vacuum or tension as in transpiration,
the pressure potential is <0. For a case of turgor
pressure the pressure potential would be >0.
Group 1
10.0
y = -25.887x + 6.4495
0.0
W%
R = 0.9664
-10.0
-20.0
0 0.2 0.4 0.6 0.8
Sucrose Concentration (m)
Figure No. 1
 y = -23.375x + 6.3734
Group 2 y = -10.714x + 10.678 Group 6 R = 0.9811
R = 0.1031
20.0 10.0
0.0

W%
W%

0.0
-10.0
-20.0 -20.0
0 0.2 0.4 0.6 0.8 0 0.2 0.4 0.6 0.8
Sucrose Concentration (m) Sucrose Concentration (m)

Figure No. 2 Figure No. 6

Group 3 Group 7
10.0 y = -12.785x + 6.0472 10.0 y = -16.021x + 4.7864
R = 0.8886 5.0 R = 0.9451

W%
5.0
W%

0.0
0.0 -5.0
-5.0 -10.0
0 0.2 0.4 0.6 0.8 0 0.2 0.4 0.6 0.8
Sucrose Concentration (m) Sucrose Concentration (m)

Figure No. 3 Figure No. 7

Group 4 Group 8
10.0 y = -8.477x + 4.7047 10.0 y = -10.55x + 4.9304
R = 0.4158 R = 0.7815
5.0 5.0
W%
W%

0.0 0.0
-5.0 -5.0
0 0.2 0.4 0.6 0.8 0 0.2 0.4 0.6 0.8
Sucrose Concentration (m) Sucrose Concentration (m)

Figure No. 4 Figure No. 8

Group 5 Group 9
20.0 y = -29.788x + 9.7058 20.0 y = -24.726x + 14.466
10.0 R = 0.959 R = 0.9475
10.0
W%
W%

0.0
0.0
-10.0
-20.0 -10.0
0 0.2 0.4 0.6 0.8 0 0.2 0.4 0.6 0.8
Sucrose Concentration (m) Sucrose Concentration (m)

Figure No. 5 Figure No. 9

 is the gas constant = 8.31J/ K.mol and is the
Group 10 temperature in Kelvin (28C +273).
20.0 y = -38.37x + 17.386
Molality of Water
R = 0.9874 Root
Group sucrose where Potential
W%

0.0 Crops W%= 0 (m) (MPa)

-20.0 Grp. 1 0.2886 -0.7220
0 0.2 0.4 0.6 0.8 Potato
Grp. 6 0.2822 -0.7059
Sucrose Concentration (m)
Grp. 2 0.4397 -1.0999
Carrot
Figure No. 10 Grp. 7 0.3275 -0.8191
As seen in Figures 1-10, it is constant for Grp. 3 0.4857 -1.2148
all groups that W% is inversely proportional to Jicama
Grp. 8 0.4906 -1.2271
sucrose concentration (m). This indicates that as
the concentration of sucrose increases, the change Grp. 4 0.5440 -1.3608
in the weight of the root crop decreases. Camote
Grp. 9 0.5509 -1.3779
Some of the plots have low coefficient of Grp. 5 0.2676 -0.6693
determination (R2) (Figures 2&4). Inaccuracies Radish
can be caused by improper blotting, errors in the Grp. 10 0.4046 -1.0121
analytical balance; leaving cylinders in the The negativity in osmotic potential is due
solutions for different amounts of time and to the fact that pure water is usually defined as
excessive drying out of the cylinders. having an osmotic potential ( ) of zero and the
The molality where the percent change in presence of solutes always make a solution have
weight (W%) is equal to 0 represents the less water than the same value of pure water.
molality at which there is no net movement of Thus, the more solute molecules present, the
water molecules and no osmotic gradient, more negative the osmotic potential is. Typical
creating an osmotic equilibrium. Since there is no values for cell cytoplasm are 0.5 to 1.0 MPa.
net osmotic movement, the turgor pressure,
The resulting solute potential in J/m3 was
which pushes the plasma membrane against the
converted into MPa by dividing 106. It is
cell wall of the plant sample, does not exist
assumed that is very small and therefore
(Steudle & Zimmermann, 2016). Therefore =
negligible, thus, = .
0.
As seen in table 1, the water potential
varies from -0.6693 to 1.3779 MPa; wherein
Table No. 1 Water Potentials of Different camote has the highest and the radish having the
Root Crops lowest. A more negative water potential indicates
that the cells of the root crop contains more
As seen in table 1, the crops have different solute. Among the root crops, the cells of the
molalities where W% = 0. The results range camote have the most solute and the cells of the
from 0.28676 to 0.5509 m; wherein camote has radish have the least solute.
the highest and the radish having the lowest.
Solute potential is defined as the pressure
The relationship of solute concentration (in that must be applied to a solution to prevent the
molality) to solute or osmotic potential is given inward flow of water through a semipermeable
by the van 't Hoff equation membrane. The flow of water stops because the
= pressure of the solution and the pressure of the
water are equal. (Martin and Hine, 2008). Solute
where is the molality (1 molal=1 x103 mol/m3 potential values are always negative. The reason
H2O), is the ionization constant = 1 for sucrose, for the negativity of solute potential is due to the
fact that pure water, which is determined to = degrees of supercooling (negative in
contain no solutes, has a solute potential sign)
equivalent to zero. By adding solutes, the solution
0.0125 = amount of water (1/80) that solidifies
would result to have less water than the same
per degree of supercooling.
value of pure water. Therefore, it can be said that
the more solute molecules present, the more
negative the Solute potential is.
Cryoscopy is a method used for the 0
determination of the lowered freezing points 0 100 200 300

Temperature (oC)
produced in liquid by dissolved substances in
order to determine the molecular weight of -0.5
solutes and various properties of solutions. With
this method, the solute potential of extracted sap
in plants can be acquired using the formula: -1

= 1.10f
-1.5
Where in 1.10 is the ratio of the osmotic Time (s)
potential of a non-iodized solution (-2.27 MPa)
and the freezing point depression of sucrose (- Figure 1.1. Depression of the Freezing Point of
2.06C) (Saupe, 2009), f or is the true Potato (Group 1)
freezing point and c is the correction factor or the
ratio between the room temperature in Kelvin
(28C +273K) and 273 K. 1.5

1
Temperature (oC)

Furthermore, cryoscopy centers upon the

phenomenon known as Depression in Freezing
Point. The freezing point of a substance is the 0.5
temperature at which a substances liquid and 0
solid form is in equilibrium. This phenomenon 0 50 100 150 200
dictates that the freezing point of a pure solvent -0.5
would be higher (less negative) than that of the
freezing point of a solution. It is also a colligative -1
Time (s)
property wherein it does not depend on the nature
of the solute molecules but rather it depends only
Figure 1.2. Depression of the Freezing Point of
on the amount of solute molecules present.
Potato (Group 6)
In determining freezing point depression
0
on tissue sap, increase of temperature would be a
direct result from the release of the heat of fusion -0.5 0 100 200 300 400
Temperature (oC)

with sudden increase as the apparent freezing -1

point (f') (Saupe, 2009). According to Reiss
(1996), after obtaining the correction for super- -1.5
cooling, the true freezing point (f) is obtained -2
using the equation:
-2.5
f = f' - 0.0125
-3
Where: Time (s)

f = true freezing point

Figure 1.3. Depression of the Freezing Point of
f' = apparent freezing point Carrot (Group 2)
 2.5 2
2 1
1.5
Temperature (oC)

Temperature (oC)
0
1
0 100 200 300
0.5 -1
0 -2
-0.5 0 100 200 300 400 500
-3
-1
-1.5 -4
-2 -5
Time (s) Time (s)

Figure 1.4. Depression of the Freezing Point of

Figure 1.7. Depression of the Freezing Point of
Carrot (group 7)
Camote (Group 4)

1 1.5
Temperature (oC)

1
Temperature (oC)
0
0 100 200 300 0.5
-1
0
-2 0 200 400 600 800
-0.5
-3 -1
Time (s) Time (s)

Figure 1.5. Depression of the Freezing Point of

Figure 1.8. Depression of the Freezing Point of
Jicama (Group 3)
Camote (Group 9)

2
1.5
1
1
Temperature (oC)

0
Temperature (oC)

0.5
0 50 100 150 200 250
-1 0
-2 0 100 200 300 400
-0.5
-3 -1

-4 -1.5

-5 -2
Time (s) Time (s)

Figure 1.6. Depression of the Freezing Point of Figure 1.9. Depression of the Freezing Point of
Jicama (Group 8) Radish (Group 5)
 1.5 Table 2. Determination of Freezing Point of
Different Root Crops
1
Temperature (oC)
Root Group Degree Apparent True Solute
0.5 Crops of Freezing Freezing Potential
Super- Point Point (f) (s)
0 cooling (f')
-0.5 0 100 200 300 400 Potato Grp. 1 -1.4oC -1oC -0.9825oC -1.1915 MPa

-1 Grp. 6 -0.7oC -0.7oC -0.6913oC -0.8384

MPa
-1.5 Carrot Grp. 2 -2.4oC -2oC -2.375oC -2.88
Time (s) MPa
Grp. 7 -1.6oC -1.4oC -1.5825oC -1.9193
MPa
Figure 1.10 Depression of the Freezing Point Jicama Grp. 3 -2.7oC -1oC -2.6875oC -3.2595
of Radish (Group10) MPa
Grp. 8 -4.2oC -2.4oC -4.17oC -5.0575
In this experiment, the concept of MPa
supercooling is applied. Supercooling is defined Camote Grp. 4 -4oC -1.2oC -3.985oC -4.833
as the process where in a liquid reaches is MPa
freezing point but does not crystallize. This Grp. 9 -0.6oC -0.3oC -0.5963oC -0.7232 MPa
process occurs because supercool liquids are Radish Grp. 5 -1.4oC -1.3oC -1.3838oC -1.6782
metastable and do not possess seeds which MPa
triggers the crystallization of the liquid sample. In Grp. -1.2oC -0.9oC -1.1888oC -1.4415
10 MPa
Figures 1.1, 1.3, 1.5, 1.6, 1.7 and 1.10, the lowest
point in the graph corresponds to the extent of the By obtaining the solute potential, we can
supercooling process happening within the sap calculate the pressure potential for each root crop
solution, the abrupt increase from the lowest with the formula:
point to its next point is termed as the apparent =
freezing point. This abrupt increase as shown in
the stated figures is because of the release of the The resulting values for the pressure potential is
heat of fusion. However, in figures, 1.2, 1.4, 1.8 shown in Table No. 3.
and 1.9 and the theoretical increase cannot be Table No. 3 Pressure Potential of Different
seen, this may be because the extent of the Root Crops
supercooling process of the root crop was not
Root Crops Group Pressure
efficiently carried out. It may also be because of
Potential
human error. Potato Grp. 1 0.4695 MPa
The extent of super-cooling of a solution is Grp. 6 0.1325 MPa
the lowest temperature value obtained. After Carrot Grp. 2 1.7801 MPa
rapid increase in temperature, which is a result of Grp. 7 1.1002 MPa
the heat of fusion, the value obtained is the Jicama Grp. 3 2.0447 MPa
apparent freezing point (f') (Saupe, 2009). Grp. 8 3.8304 MPa
Camote Grp. 4 3.4722 MPa
Using the equation, f = f' - 0.0125 ,
Grp. 9 -0.6547 MPa
the true freezing point for each of the root crops
Radish Grp. 5 1.0089 MPa
were obtained. The values of true freezing point
Grp. 10 0.4294 MPa
for each root crop were used to calculate their
solute potentials respectively. All the obtained
results are presented in Table No. 2. Possible sources of error in determining
the true freezing point may be because of the state
of the ice used. There were periods within the
experiment where in other groups used melted ice
while others used new ice.
Conclusion Answer: During equilibrium, the sign of the
pressure potential of tissues is 0. At incipient
The water potentials of Potato ranges
plasmolysis, the pressure potential will still be 0
from -0.70 to -0.72 MPa, Carrot ranges from -
and during transcription, particularly in xylem
0.82 to -1.10 MPa, Jicama ranges from -1.21 to -
cells, there will be a negative pressure potential.
1.23 Mpa, Camote ranges -1.36 to -1.38 MPa, and
Radish ranges from -0.67 to -1.01 MPa. Camote
has the lowest water potential and highest solute
4. What causes supercooling? Why is the
concentrations while Radish has the highest water
lowest temperature reached not considered
potential and lowest solute concentrations. Upon
the freezing point of the sap solution?
obtaining the water potential and solute potential,
they were used to calculate the pressure potential Answer: Supercooling, also known as
for each of the root crops. Pressure potential for undercooling, is the process when a liquid
camote ranges from -0.6547 MPa to 3.4722 MPa, reaches below its freezing but does not become
for potato it ranges from 0.1325 MPa to 0.4695 solid. This occurs because supercooled liquids
MPa, for carrot it ranges from 1.1002 MPa to that are metastable do not possess seeds that may
1.7801 MPa, for Jicama it ranges from 2.0447 trigger the crystallization. The reason why the
MPa to 3.8304 MPa and for Radish it ranges from lowest temperature of the sap solution is not
0.4294 MPa to 1.0089 MPa. considered because sap solutions usually reach -
7 to -16 degrees Celsius, once they reach below
Application their supercooling limit, ice forms and the plant
1. Why is the osmotic potential always dies (Korner, 2012).
negative in sign?
Answer: The value of osmotic potential is always 5. Do plants with watery sap have higher or
negative in sign since the presence of solutes, lower water potential? Explain your answer
which water binds to through hydrogen bonding using actual computed values.
consumes some of the potential energy available
in water. Thus, it will always make a solution Answer: Based on the table presenting the
have less water than the same value of pure water. computed values for water potential for each root
crops, it can be inferred that the more watery sap
a root crop has the lower its water potential. This
2. How do dissolved solutes contribute to the is based on the principle that the higher the
water potential of the cell? concentration of solute a root crop has the more
negative its water potential shall be.
Answer: Water potential is defined by its
equation: = + + , where in stands Molality of Water
Root
for the concentration of the solutes or the osmotic Group sucrose where Potential
Crops
potential and it always has a negative sign W%= 0 (m) (MPa)
because of the presence of solutes. Solutes can be Group 1 0.2886 -0.7220
dissolved by water because water can bind Potato
through them via hydrogen bonding, this process Group 6 0.2822 -0.7059
consumes some of the potential energy available Group 2 0.4397 -1.0999
in water and since it becomes more negative as Carrot
more dissolved solutes are added this also lowers Group 7 0.3275 -0.8191
the water potential of the cell. Group 3 0.4857 -1.2148
Jicama
Group 8 0.4906 -1.2271
3. What is the sign of the pressure potential of Group 4 0.5440 -1.3608
tissues at equilibrium? (+), (0), or (-)? At Camote
incipient plasmolysis? During transpiration? Group 9 0.5509 -1.3779
Group 5 0.2676 -0.6693
Radish
Group 10 0.4046 -1.0121
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