TRANSPORT OF MATERIALS ACROSS CELL MEMBRANES &
PLANT-CELL WATER RELATIONS
Alcantara. Catindig. Ignacio. Kim.GROUP 2
DIFFUSION OF SELECTED PLANT PIGMENTS
4 plant specimens
A B C D
dH2O dH2O+
H2O bath
veg. oil heated veg. oil
plant specimen
*a total of 16 test tubes were used
Methodology
Set aside for 30 minutes
Shake test tubes
Compare color intensities. Record results.
Methodology
Results
Test tube 1 (w/ dH2o) +++
Test tube 2 (w/ heated dH2o) ++++
Test tube 3 (w/ veg. oil) +
Test tube 4 (w/ heated veg. oil) ++
Bixa orellana
Results
Test tube 1 (w/ dH2o) +
Test tube 2 (w/ heated dH2o) ++
Test tube 3 (w/ veg. oil) +++
Test tube 4 (w/ heated veg. oil) ++++
Zingiber officinale
Results
Test tube 1 (w/ dH2o) +
Test tube 2 (w/ heated dH2o) ++
Test tube 3 (w/ veg. oil) +++
Test tube 4 (w/ heated veg. oil) +++
Solanum toberosum
Results
Test tube 1 (w/ dH2o) ++
Test tube 2 (w/ heated dH2o) +++
Test tube 3 (w/ veg. oil) +
Test tube 4 (w/ heated veg. oil) +
Allium cepa
Discussion
Diffusion: directed movement of molecules from a region of high concentration to a region of lower concentration random thermal motion
Affected by: Concentration and size of diffusing particles
Discussion
Bixa orellana contain the pigments bixin and orelline Carotenoid pigments Lipid-soluble due to long hydrocarbon chain
Zingiber officinale contain flavonoids (quercetin, rutin,
catechin, epicatechin, kaempferol and naringenin)
Lipid-soluble due to the ring-like carbon structures.
Discussion
Red Onion Anthocyanin: water-soluble Quercetin: lipid-soluble
Potato skin Contains carotenoid pigments (neoxanthin,
violaxanthin and lutein) Lipid-soluble
Discussion
Bixin and orelline were able to diffuse much faster than the others
Carotenoids are able to reach high concentrations within chromoplastids and may actually form crystals
Large amount of bixin and orelline increased the rate of their diffusion throughout the medium
OSMOSISCELL CHANGES IN PLASMOLYSIS
Osmosis: Cell Changes in Plasmolysis
OSMOSIS Diffusion of water across a semi-
permeable membrane
Osmosis: Cell Changes in Plasmolysis
1
•A Tradescantia spathacea leaf was obtained and strips of its lower epidermis were prepared using a blade.
2
•A wet mount was made using the lower epidermis and the cells were observed under the microscope.
3
•Water was drawn off the slide with tissue paper and was replaced with a drop of 5% NaCl.
Osmosis: Cell Changes in Plasmolysis
4
•The cells were again observed under a microscope and changes were noted.
5
•The procedure was repeated using white onion and then apple skin.
Discussion of Results
Tradescantia spathacea
Wet mount 5% NaCl
Discussion of Results
5% NaClWet mount
Allium cepa
Discussion of Results
5% NaClWet mount
Malus
Discussion of Results
Turgid cell happens when cell is hypotonic to the
surrounding solution optimal for plants
Plasmolyzed cell happens when cell is hypertonic to the
surrounding solution; plasma membrane lysis may cause cell death cell wall still intact
Discussion of Results
Anthocyanin water-soluble pigment discoloration in plasmolysis
Conclusion
Osmosis is the diffusion of water through a semi-permeable membrane and this can be observed using different epidermal cells with pigments
Cells in hypotonic solutions become turgid and cells in hypertonic solutions become plasmolyzed as water goes in and out of the cell, respectively.
FACTORS AFFECTING INTEGRITY OF CELL MEMBRANE
Methodology
Red apple peel
In test tubes
A: Distilled + Room
Temp
B: Distilled +
Refrigerator
C: Distilled + 60°
Under the microscop
e
D: 50% Chlorofor
m
E: 50% Acetone
F: 0.1M NaOH
G: 0.1M HCl
Results
Test Tube Intensity of Color
A (room temp.) +++
B (refrigerator/cold) ++
C (water bath/ 60C) +
D (Chloroform) +
E (Acetone) ++
F (NaOH) +++
G (HCl) ++++
Discussion
Red violet pigment in apples: ANTHOCYANIN
Found at the vacuole
Too big to exit cell membrane and tonoplast
Discussion
Heat: Denatures proteins; destroys membrane
Cold: fatty acid tails rigid; less permeability
Organic Solvents interact with bilayer causing disruption of membrane
Low and High pH: destroys tertiary and quaternary structure of pigments
DETERMINATION OF SOLUTE CONCENTRATION OF CELLS
(PLASMOLYTIC METHOD)
10Drops of sucrose solution + Tradescantia spathacea
epidermal strips(0.1M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M, 0.8M, 0.9M and 1.0M)
30 minutes
Methodology
Wet mount
Observed under the microscope
The number of PLASMOLYZED and UNPLASMOLYZED cells were recorded as well as the concentration that caused INCIPIENT PLASMOLYSIS.
The OSMOTIC POTENTIAL value was also calculated.
Methodology
Discussion of Results
OSMOSIS: diffusion of water across a semi-permeable membrane
Water potential (Ψw) Important in determining the direction of
osmosis High to low Ψw
PLASMOLYSIS: shrinking of a cell due to water loss happens when a cell is submerged in a
hypertonic solution
Source: http://www.excellup.com/interbiology/planttransportquestion.aspx
Discussion of Results
The cell wall is permeable to water and sucrose.
The plasma membrane is permeable to water but not to sucrose.
Sucrose + Water
Discussion of Results
Sucrose Concentrati
on (M)
Osmotic Potential
(bars)
Plasmolyzed Cells (#)
Unplasmolyzed Cells
(#)
Total # of Cells
Counted
% Plasmolyze
d
0.1 -2.5 6 145 151 3.97
0.2 -5.0 20 174 194 10.31
0.3 -7.5 41 84 125 32.8
0.4 -10.0 90 105 195 46.15
0.5 -12.5 99 88 187 52.94
0.6 -15.0 186 85 271 68.63
0.7 -17.5 112 83 195 57.44
0.8 -20.0 76 54 130 58.46
0.9 -22.5 89 61 150 59.33
1.0 -25.0 172 73 245 70.20
Data showing the osmotic potential, number of plasmolyzed and unplasmolyzed cells, total number of cells counted, and
the percentage of plasmolyzed cells found under the microscope for each concentration of sucrose solution used.
Using the data from petri dish 1,
Sucrose Concentrati
on (M)
Osmotic Potential
(bars)
Plasmolyzed Cells (#)
Unplasmolyzed Cells
(#)
Total # of Cells
Counted
% Plasmolyze
d
0.1 -2.5 6 145 151 3.97
0.2 -5.0 20 174 194 10.31
0.3 -7.5 41 84 125 32.8
0.4 -10.0 90 105 195 46.15
0.5 -12.5 99 88 187 52.94
0.6 -15.0 186 85 271 68.63
0.7 -17.5 112 83 195 57.44
0.8 -20.0 76 54 130 58.46
0.9 -22.5 89 61 150 59.33
1.0 -25.0 172 73 245 70.20
Data showing the osmotic potential, number of plasmolyzed and unplasmolyzed cells, total number of cells counted, and
the percentage of plasmolyzed cells found under the microscope for each concentration of sucrose solution used.
percentage of plasmolyzed cells increased as the concentration of sucrose in the solution increased.
sucrose concentration of 0.6M - 68.63% of plasmolyzed cells
Discussion of Results
INCIPIENT PLASMOLYSIS osmotic potential of the cell is the same as
the solution’s the protoplast just fills the cell volume and
neither exerts pressure to the cell wall nor withdraws from it
50% of plasmolyzed cells
Discussion of Results
Sucrose Concentrati
on (M)
Osmotic Potential
(bars)
Plasmolyzed Cells (#)
Unplasmolyzed Cells
(#)
Total # of Cells
Counted
% Plasmolyze
d
0.1 -2.5 6 145 151 3.97
0.2 -5.0 20 174 194 10.31
0.3 -7.5 41 84 125 32.8
0.4 -10.0 90 105 195 46.15
0.5 -12.5 99 88 187 52.94
0.6 -15.0 186 85 271 68.63
0.7 -17.5 112 83 195 57.44
0.8 -20.0 76 54 130 58.46
0.9 -22.5 89 61 150 59.33
1.0 -25.0 172 73 245 70.20
Data showing the osmotic potential, number of plasmolyzed and unplasmolyzed cells, total number of cells counted, and
the percentage of plasmolyzed cells found under the microscope for each concentration of sucrose solution used.
Concentration where incipient plasmolysis occurred
is 0.5M with a 52.94% of plasmolyzed cells.
Discussion of Results
Discussion of Results
The osmotic potential (Ψs), in bars, of the sucrose solutions were computed using this formula:
where:m = concentration of the solute expressed as molality (moles solute/ kg H2O)i = ionization constantR = gas constant (8.314 J/mol∙K)T = absolute temperature (C + 273)
Sample computation:Osmotic potential of a 0.1 M sucrose
solution
Ψs = -(0.1 mol/L)(1)(8.31 J/K-mol)(300K)Ψs = -249.3 J/L (0.01 bars/ 1 J/L) = -2.493 barsΨs ~ -2.5 bars
Discussion of Results
Sucrose Concentrati
on (M)
Osmotic Potential
(bars)
Plasmolyzed Cells (#)
Unplasmolyzed Cells
(#)
Total # of Cells
Counted
% Plasmolyze
d
0.1 -2.5 6 145 151 3.97
0.2 -5.0 20 174 194 10.31
0.3 -7.5 41 84 125 32.8
0.4 -10.0 90 105 195 46.15
0.5 -12.5 99 88 187 52.94
0.6 -15.0 186 85 271 68.63
0.7 -17.5 112 83 195 57.44
0.8 -20.0 76 54 130 58.46
0.9 -22.5 89 61 150 59.33
1.0 -25.0 172 73 245 70.20
Data showing the osmotic potential, number of plasmolyzed and unplasmolyzed cells, total number of cells counted, and
the percentage of plasmolyzed cells found under the microscope for each concentration of sucrose solution used.
Conclusion
As solute concentration increased, osmotic potential became more negative along with the water potential
% of plasmolyzed cells also increased as water potential became more negative Water diffuses to a region with a more negative water
potential To equilibrate the concentration of water inside of cell
to that of the surrounding solution, water moved out of the cell
ESTIMATION OF THE WATER POTENTIAL OF STORAGE TISSUE
(VOLUME CHANGE METHOD)
Methodology
1
•Eleven sets of five potato cylinders (each potato cylinder 1cm long) were cut off from a large potato and immediately placed in 50 mL beakers
2
•20-ml of one concentration of sucrose solution (0.1 M-1.0 M, with 0.1 graduations) were placed in the 10 separate beakers respectively.
3
•The remaining beaker contained 20 mL distilled water
Methodology
4
•The fresh weights of each set were recorded. The potato cylinders were removed after 90 minutes and weighed again.
5
•The difference between the initial and final weights were divided by the initial weight, and then multiplied by 100 to get % weight change.
∆weights of the potato cylinders=caused by the presence of sucrose (this stimulated the cells to generate an osmotic potential (Ψs))
Discussion of Results
Osmotic potential reduces the free energy of the system.
The effect of osmotic potential is countered by hydrostatic pressure.
Discussion of Results
Sucrose Contentration
(M)
Initial Weight(go)
Final Weight (g)
∆ Weight (g - go)
% ∆ Weight
0 2.7930 2.8600 0.0670 2.40
0.1 2.8373 2.8325 0.0048 0.169
0.2 2.7395 2.5479 0.1916 6.99
0.3 2.6992 2.3307 0.3685 13.65
0.4 2.6900 2.0258 0.6642 24.69
0.5 2.8865 2.1195 0.7670 76.70
0.6 2.8773 3.0325 0.1552 5.39
0.7 2.9200 3.0635 0.1435 4.91
0.8 2.9564 3.0940 0.1376 4.65
0.9 2.8681 2.9964 0.1354 4.72
The initial, final, and change in weights and the Percent Weight Change of potato cylinders placed in different concentrations of sucrose solutions for 90 minutes.
0 0.2 0.4 0.6 0.8 1 1.20
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
Sucrose Concentration (in M, moles/L)
Percent Change in Weight (g)
Fig 5. Plot of Percent Change in Weight (in grams) vs. Sucrose Concentration (in M, moles/L).
Discussion of Results
Experimental data: failed to present the expected trend and failed to show the concentration of sucrose where there is 0% ∆ in weight
Theoretical data would show that the higher the concentration of sucrose, the higher the percent change in weight.
Conclusion
Theoretically, the sucrose concentration between 0.2-0.3M should have registered the zero percent change in weight.
Discussion of Results