PLANT PHYSIOLOGY PRACTICUM

CELL MEMBRANE CHEMICAL COMPOSITION AND FACTORS

AFFECTING PERMEABILITY

ALIVIA ZULKARNAIN

2110422037

Group 3 KBI

PLANT PHYSIOLOGY PRACTICUM

BIOLOGY DEPARTMENT

MATHEMATIC AND SCIENCES FACULTY

ANDALAS UNIVERSITY

2022
 I. INTRODUCTION

The membrane is defined as a porous medium, in the form of a thin film, which is
semipermeable which serves to separate particles of molecular size (species) in a solution
system. Species that have a size larger than the membrane pore will be retained while the
species with a smaller size will pass through the membrane pore (Kesting, RE, 2000).
The plasma membrane is the boundary of life, the boundary that separates the
living cell from its dead surroundings. This extraordinary thin layer of only about 8 nm
thick requires more than 8000 plasma membranes to control traffic into and out of the
cells it surrounds. Like all biological membranes, the plasma membrane has selective
permeability, etc. it allows some substances to pass through it more easily than others.
One of the earliest episodes in the evolution of life may have been the formation of a
membrane that confines a solution that has a different composition than the surrounding
solution, but which is still capable of absorbing nutrients and removing waste products.
The ability of the cell to distinguish these chemical exchanges from that of its environment
is fundamental to life, and it is the plasma membrane that makes this selectivity possible.
(Campbell, et al., 2002).
Cells have an outer layer that separates the nucleus from the environment. The
outermost layer is called the cell membrane. The main building blocks of cell membranes
are lipids and proteins (lipoproteins). The cell membrane consists of a phospholipid
bilayer. Phosphate as the head is hydrophilic (likes water) and fat as the tail is hydrophobic
or rejects water (Yusa and Manikam, 2016).
Cell membranes are selectively permeable or semipermeable because only certain
ions, molecules, and compounds can pass through. In animal and human cells, the cell
membrane is located on the outermost part, while in plants, the cell membrane is
surrounded by a cell wall. The plasma membrane is composed of lipids (phospholipids),
proteins, and carbohydrates (Irnaningtyas, 2016).
Cell membrane is a thin structure consisting of phospholipids and proteins, where
phospholipids are the basic structure of cell membranes because they consist of
hydrophobic and hydrophilic parts that are close together to form two layers. The main
functions of this plasma membrane are selective permeability and solvent transfer, dextron
transfer, and oxidative phosphorylation, excretion of hydrolytic exoenzymes, generating
receptors and other proteins, producing enzymes and carrying molecules for the
biosynthesis of DNA, cell wall polymers and membrane lipids. If the cell membrane is in
a liquid state, it can help the bacterial cell to grow normally and if there is damage to this
structure, there will be interference with the cell, resulting in death (Jawetz, et al., 2001).
 II. METHODS

2.1 Time and Place

The practicum about Water as a Plant Component was held on Monday, October 3rd ,
2022, 8:00 – 12:00 WIB at the Education Laboratory IV, Department of Biology, Faculty
of Mathematics and Natural Sciences, Andalas University, Padang.
2.2 Tools and Materials
The tools used in this practice are 1000 ml beaker, drill to make shaped pieces cylinder
with a diameter of 1 cm, spectrophotometer and cuvette, thermometer, freezer, test tube,
and razor blade. Materials that needed in this practicum are tubers of the type of tuber
plant, leaves of the Rhoe discolor plant, distilled water, HCl solution (0.025 N), KOH
(0.025 N), Acetic acid (0.025 N), NH4OH (0.025 N).
2.3 Procedure
2.3.1 Effect of Temperature and Chemical Compounds on Cell Membrane Permeability
Choose one large tuber, wash it thoroughly with tap water if necessary, brush it. Then with
the help of a drill with a diameter of 1 cm (the center is hollow), cut 12 cylindrical shapes
from the same tuber with a cut thickness of 3 cm. washed all pieces of tuber under running
water (tap water) for 10-15 minutes to remove pigment on the surface.
2.3.1.1 Heat Treatment
Prepare a water bath by filling 2/3 part of a 1000 ml beaker with water, and heat it over a
fire or hot plate. Also prepared are 5 test tubes filled with 15 ml of distilled water. Each 1

piece of tuber was put into water with different temperatures (room temperature, 40oC,

55oC, 70oC, and 80oC). The pieces are soaked for 1 minute. After 1 minute, remove the
tuber pieces with tweezers, then put them in a test tube containing aquadest. The tuber
pieces are left for 1 hour, after that shake the test tube and pour the marinade into the
cuvette. The absorbance value was measured at a wavelength of 525 nm on a
spectrophotometer. Repeat the same steps for each water bath (5 heat treatments). If the
solution after immersion for 1 hour is too thick (high pigment concentration), all samples
are diluted with distilled water (1:1) and the measurement is repeated.
2.3.1.2 Cold Treatment
The tuber pieces at the beginning of the experiment were put in the freezer so that they
were frozen. The frozen tubers were then washed quickly with tap water and put into a
test tube containing 15 ml of water. As a control, place one piece of unrefrigerated potato
tuber in a test tube with 15 ml of water. After incubation for 1 hour, measure the relative
amount of pigment in the immersion solution with a spectrophotometer. If part A is
diluted, then part B is also diluted.
2.3.1.3 Treatment with Chemical Compounds
Place one cylindrical piece of potato tuber each into 15 ml of a solution of methanol,
acetone, and tertiary butyl alcohol. As a control, put one piece of potato tuber into 15 ml
of distilled water. Then incubated for 1 hour and measure the absorbance using a
spectrophotometer.
2.3.2 Permeability of Living Tissue in Acid and Base Solutions
Prepare an incision of the lower epidermis of Rhoe discolor and place it in distilled water.
Prepare the test solution (distillate water, HCl, KOH, acetic acid and NH4OH) into the
tube reaction 10 mL each and label them. Place two incisions of epidermal tissue in
distilled water, two incisions in KOH and six incision in NH4OH. Record the time it takes
for the color to turn blue after immersion. After the color turns blue in the NH4OH
solution, transfer the four incisions into distilled water. From distilled water, transfer the
two incisions into the acetic acid solution and the two incisions into HCl. Record the time
for the discoloration to occur in the tissue. When the color change is complete, transfer
the tissue incision into distilled water and return it into the NH4OH solution. Record the
time it takes for the color to change again. Perform procedures 4 and 5 three to five times
and calculate the average time required to color changes under acidic and basic conditions.
 III. RESULTS AND DISCUSSION

3.1 Results
Table 1. Absorbance Value in Hot and Cold Temperature Experiments
Temperature Absorbance value
Room temperature (27oC) 0,222
40 0,025
55 0,056
70 0,009
80 0,0089
Cold (freeze) 0,018
Control 0,095

Table 2. Absorbance Value in Chemical Compound Experiments

Solution Absorbance value
Aquadest (control) 0,200
Methanol 0,075
Acetone 0,015
Buthanol 0,008

Table 3. Color Changing in Rhoeo discolor Leaf for Minutes

Solution Color changing Time
Aquadest Not really changed 30 minutes
KOH Becomes blue 1 minutes
Acetic acid Becomes transparent 27 minutes
HCl Becomes red 12 minutes
3.2 Discussion
3.2.1 Measurement of Water Content in Plant Tissues
From the results of the experiment, the higher the given temperature, the greater the
absorbance value. Because the temperature is too extreme for the membrane to withstand,
the result is that the membrane cannot withstand temperatures that are too high or too low.
The higher the temperature, the more damaged the membrane will be. As a result, more
and more cell contents come out. If the temperature is too high, the protein will experience
denaturation and cause the contents in the cell to come out because the proteins that make
up the cell membrane are damaged. The difference in permeability depends on the size of
the molecules that pass through and is determined by the size of the membrane pores
(Niemetz, 2006).
The difference in permeability is very dependent on the size of the molecules
passing through and is determined by the size of the membrane pores. But on the plasma
membrane of living cells the size of the molecule has no effect, this is due to the
relationship between the solubility of substances in one of the membrane components.
The plasma membrane has selective permeability, i.e. it allows some substances to pass
through it more easily than others. The ability of the cell to distinguish these chemical
exchanges from that of its environment is fundamental to life, and it is the plasma
membrane that makes this selectivity possible. (Campbell, 2002).
According to Bonner (1961), each application of different chemicals affects
different levels of permeability. The administration of methanol affects the level of cell
membrane damage which is high compared to treatment using a high level of cell
membrane damage compared to treatment using acetone or benzene. Methanol is a polar
alcohol compound that can dissolve organic compounds such as cell membranes. The
dissolved membrane then loses its turgidity and causes the cell contents to escape. Acetone
is an excellent solvent for a wide variety of organic compounds, leaving the cells almost
identical to that of methanol. Benz is an aromatic compound that is insoluble in air and is
in the form of an emulsion.
In the experimental observation of color changes in leaf incisions of Rhoeo
discolor with 4 different solutions, the results obtained in aquadest solution, the leaves did
not have too large a color change, even after 30 minutes. In leaf incisions soaked with
KOH solution, the color of the leaf incision changes to blue within 1 minute. In leaf
incisions soaked with acetic acid solution, the color of the incision changes to transparent
slowly within 27 minutes. In leaf slices soaked with HCL solution, the color of the leaf
incision changed to red within 12 minutes.
According to Ratnasari (2016), in his research it was known that the ethanolic
extract of Rhoeo discolor leaves contained a purple pigment, namely anthocyanins which
have the characteristic of changing color with every change in pH so that they have the
potential as an alternative acid-base indicator, and has been proven by Padmaningrum
(2011), the extraction results Rhoeo discolor leaves with alcohol solvent experience a
pink-green yellow color change on acid-base titration. This proves that the extraction
results of Rhoeo discolor leaves can be used as an alternative acid-base indicator to replace
synthetic indicators.
The use of acid-base indicator paper from Rhoeo discolor leaf extract has
advantages compared to red and blue litmus. The test results of acid-base indicator paper
from Rhoeo discolor leaf extract can distinguish between a strong acid solution and a weak
acid and a strong base solution with a weak base, while red and blue litmus are only able
to distinguish an acidic or basic solution.
Acid-base indicator paper from Rhoeo discolor leaf extract in a strong acid
solution is peach in color and in a weak acid solution is pink, while in a bottled green
strong base solution and a weak base solution is dark green. Meanwhile, red litmus when
dipped in a solution of a strong acid or a weak acid will turn red (fixed), and if it is dipped
in a solution of a strong or weak base, it will turn blue. Meanwhile, blue litmus when
dipped in a solution of a strong acid or weak acid is red, and if it is dipped in a solution of
a strong base or a weak base it remains blue (Latih, 2017).
Compounds that play a role in changing the color of natural indicators are
anthocyanins which are also secondary metabolites of the flavonoid group and are
naturally water-soluble pigments that have the ability to react both with acids and bases.
Anthocyanins are red in acidic media, and turn purple and blue in alkaline media
(Winarno, 1992).
 IV. CONCLUSION AND SUGGESTION
4.1 Conclusion
From the practicum that has been carried out, it can be concluded that:
1. The higher the temperature, the higher the absorbance value because of the large
number of cell contents that come out.
2. Methanol and acetone are alcohol compounds that can dissolve organic
compounds such as cell membranes.
3. Rhoeo discolor is a plant that can be used as an indicator of acid and base due to
its anthocyanin content. KOH solution is a strong base, HCL is a strong acid, and
acetic acid is a weak acid compound.

4.2 Suggestion

Practitioners are expected to be more careful and thorough in carrying out the practicum
so that the results obtained are more optimal and in accordance with the results that should
be.
 REFERENCES

Bonner, J., Huang, R. C. C., & Maheshwari, N. (1961). The physical state of newly
synthesized RNA. Proceedings of the National Academy of Sciences, 47(10), 1548-
1554.

Campbell, Neil. 2002. Biologi. Jakarta : Erlangga.

Campbell, Neil. A., Jane B. Reece, Lawrence G. Mitchel. 2002. Biologi Edisi kelima Jilid
II. Penerbit Erlangga: Jakarta.

Irnaningtyas, (2016), Biologi Untuk SMA/MA Kelas X Kurikulum 2013, Erlangga,

Jakarta.

Jawetz, E., Melnick, J. L., Adelberg, E. A., 2001, Mikrobiologi Kedokteran, Edisi XXII,
diterjemahkan oleh Bagian Mikrobiologi Fakultas Kedokteran Universitas
Airlangga, 205-209, Penerbit Salemba Medika, Jakarta.

Kesting, R. E. 2000. Synthetic Polymeric Membranes. New York: McGraw-Hill Book

Company.

Latih, Garin Puspa. 2017. Pengaruh Variasi Pelarut Daun Rhoeo discolor Terhadap
Stabilitas Kertas Indikator Asam Basa Alternatif. Surakarta.

Niemetz, Christa. 2006. Plasma Membran of Beta vulgaris Storage roots Shows High
Water Channel Activity Regulated by pH. Journal of Experimental Botany 57 : 3.

Padmaningrum, Regina Tutik. 2011. Karakter Ekstrak Zat Warna Daun Rhoeo discolor
Sebagai Indikator Titrasi Asam Basa. Prosiding Seminar Nasional Penelitian,
Pendidikan dan Penerapan MIPA, Fakultas MIPA, Universitas Negeri Yogyakarta.
Ratnasari, Sinta. 2016. “Studi Potensi Ekstrak Daun Adam Hawa (Rhoeo discolor)
Sebagai Indikator Titrasi Asam-Basa”. Jurnal Chimica et Natura Acta Vol.4 No.1,
April 2016: 39- 46.

Winarno, F.G. (1992). Kimia Pangan dan Gizi. Edisi Keenam. Jakarta: Gramedia.

Yusa & M. B. S. Manikam. 2016. Buku Siswa Aktif dan Kreatif Belajar Biologi. Bandung:
Grafindo Media Pratama.
 ATTACHMENT

Fig. 1 rhoe discolor with KOH treatment Fig. 2 rhoe diacolor before given
treatment

Fig 3. Rhoe discolor with aquadest Fig. 4 Rhoe discolor with acetic acid
treatment treatments

Fig 5. Rhoe discolor with aquadest Fig 6. Measurements of absorbant value

treatment with spectrophotometer