Onion Root Tip Mitosis

E. J. Gallegos, S. D. Seva, I. Clutario, N. A. De los Santos

STEM 12 - Franklin

Legazpi City Science High School

Bitano, Legazpi City
Senior High School Department
Science, Technology, Engineering, and Mathematics

Date Performed: September 25, 2023

ABSTRACT
Mitosis is a cell process where a parent cell reproduces into two genetically identical
daughter cells. It is essential for one’s growth, repair, and development. Mitosis has two major
stages: Cytokinesis which is the cytoplasm division and Karyokinesis which is the nucleus
division. Under Karyokinesis, there are four main stages including Prophase, Metaphase,
Anaphase, and Telophase. It may happen to both animal and plant cells. In animal cells,
mitosis occurs in almost all body cells except in the gamete or sex cells in the sex organs. In
plant cells, it usually occurs in the root tips, shoot tips, and cambium. This laboratory
experiment aimed to observe, identify, and describe the different phases and stages of mitosis
happening in onion root tip cells. Onion root tip was utilized in this active because of the
meristematic cells that are located in the tip of the roots. This can be considered as the most
suitable plant specimen to understand the different stages of mitosis. Another reason is that
onion is a monocot plant that possesses large chromosomes that can be easily visualized.
Prior to the actual conduct, the specimen used was already prepared by growing the onion
root tips for four days. The mitosis was then identified with the roots from the onion. It was
squashed gently for the cell layer to be straightened out. The specimen was studied in a
microscope under different magnifications to obtain a distinct structure and clear image of the
cells. It was found that the root tip of the onion was undergoing the mitosis process. Most of
the cells were in the interphase while some were already proceeding to the next phases such
as prophase, metaphase, anaphase, telophase, and even cytokinesis. The data collected
were qualitative data aligned to the objectives. Different stages were identified, and results
were gathered. Through the image under the microscope, the stages of mitosis were
differentiated from each other to determine the unique features of each process.

Keywords: Mitosis, Cytokinesis, Karyokinesis, Animal Cells, Plant Cells, Onion Root Tip

INTRODUCTION (ELIJAH) exits interphase, goes through mitosis, and

The cell cycle is the sequence of divides completely. The daughter cells that
events that occur in a cell as it grows and arise enter their own interphase and begin
divides. A cell spends the majority of its a new round of the cell cycle. According to
time in what is known as interphase, where Schafer KA (1998), cyclin-dependent
it grows, copies its chromosomes, and kinases (CDKS), which are complex with
prepares for cell division. The cell then the cyclin proteins, are important to this
process. These proteins govern the the genes required to develop and divide
passage of the cell through the phases of again (McIntosh, 2016).
the cell cycle and are regulated by a variety Meiosis, the second kind of cell
of proteins, including p53, p21, p16, and division, guarantees that people have the
cdc25. Because of the cell cycle's same number of chromosomes from
pervasiveness, it ought to be included in generation to generation. It is a two-step
the design and interpretation of research in procedure that decreases the number of
a wide range of fields. chromosomes by half—from 46 to 23—in
Checkpoints aid in ensuring that order to generate sperm and egg cells.
cell cycle events take place in the proper When sperm and egg cells combine during
order. Mammalian cell studies revealed conception, each contributes 23
inhibitors of cyclin complexes and cyclin- chromosomes, resulting in a 46-
dependent kinases as cell cycle checkpoint chromosome embryo. Meiosis also allows
components, providing the first glimpse of for genetic variety by rearranging genes
the molecular processes that prohibit cells when the cells divide (Medicine plus).
with damaged DNA from reproducing their Meiosis, the antithesis of fertilization, is the
DNA. The extent to which checkpoints unique genetically programmed mode of
block cell cycle progression in embryos nuclear division associated with a halving
reflects the relative strength of inhibitory of the chromosome number in sexually
checkpoints and the machinery propelling reproducing eukaryotes. It thus represents
the cell cycle forward (KA, Murray 1994). a key cellular and developmental pathway
Mitosis is a necessary step for life. in the life of an organism, according to
A cell copies all of its components, John, B. (1990)
including its chromosomes, and separates In general, the two processes are
to generate two identical daughter cells comparable in terms of the steps the cell
during mitosis. Because this process is so goes through, but the final goals are
important, the phases of mitosis are distinct. Meiosis is important for genetic
meticulously regulated by specific genes. variety and the continuance of reproduction
When mitosis is not properly controlled, it in living organisms, whereas mitosis is
can lead to health issues such as cancer., essential for cell growth and development.
in accordance with an article from Medicine Furthermore, meiosis is responsible for the
plus. correction of genetic abnormalities.
All eukaryotic cells prepare for cell Understanding other biological phenomena
division by producing a "mitotic spindle"—a gets simpler with the two cell division kinds
bipolar machine consisting of microtubules offering knowledge on the mechanics of
(MTs) and other related proteins. This living creatures.
mechanism arranges the previously In order to develop a general
duplicated DNA in such a way that one understanding of the fundamental
copy of each chromosome connects to processes involved in life’s systems, this
each end of the spindle. Controlled MT activity tried to achieve the following
dynamics, as well as the activities of objective:
numerous motor enzymes, are required for 1) Using a microscope, observe
spindle formation and operation. Spindle mitosis--cell division--in onion root
movements divide the duplicated tip cells.
chromosomes into two different sets, which 2) Identify the mitotic phases seen.
are subsequently transferred to opposing 3) Explain the steps of mitosis.
ends of the cell. As a result, the two cells
formed during cytokinesis possess all of METHODOLOGY
 Materials
Drawing
The main material used in thiswith label
experiment was the onion in which its rootcell parts
tips were grown by utilizing a plastic bottle
and toothpicks. The specimen which was Description
the onion root tips was held in place with
the aid of glass slides and cover slips. The Guide questions were provided to help
rest of the materials were available in the understand and comprehend the
laboratory area with the assistance of the experiment’s concept:
biology teacher. Blades, scalpels, and a) Why is onion root tip selected for a
forceps were used to remove the onion's study of mitosis?
roots and place the onion on particular b) Identify the onion cell division
items in the next procedure. The onion stages viewed in onion root tip.
roots were heated and held in a vial and c) What are the stages of mitosis?
watch glass. A burner was specifically used d) Differentiate the stages of mitosis.
for the task of heating up the onion roots
using the vial. Other materials that Procedures
involved the use of liquid components were The following steps were followed
Aceto alcohol, Safranin o Stain, and to complete the experiment to ensure a
distilled water. smooth process.

Instrument/Tool Preparation of Onion Root Tips

The researchers were provided Prior to the lab experimentation, the
with a data table for the results for clear onion root tip was prepared. Half of the
presentation. The data table shows the plastic bottle, which will hold the onion for a
observed pictures, the drawing with labels, few days (3-4 days), was filled with clean
and the description for each phase. water. When only the bottom portion of the
onion was in contact with the water,
Table 1. Template for the Data Table toothpicks were used to support the onion.
The root tips were already prepared
Karyokinesis
Interphase together with the supplies that were on
Prophase Metaphase
hand in the lab before the experiment
Observed began. The Light Compound Microscope's
(pictures) reflection of light was prepared and
adjusted alongside the preparation of other
materials.
Drawing
with label
cell parts Experimentation
Using a scalpel, the researchers
Description gently snipped out parts of root, specifically
the tip, from the onion and placed them in
a petri dish. The root tips were then placed
Karyokinesis in a vial and submerged in Aceto Alcohol, a
Cytokinesis combination of glacial acetic acid and
Anaphase Telophase
ethanol. The root tips were soaked in the
Observed petri dish containing Aceto Alcohol for 5
(pictures) minutes before placing it above a burner for
another 5 minutes. Following that, the root
tips were carefully taken out from the (not a identify the stages of mitosis in onion root
vial), rinsed with distilled water, and dyed tip using the documentation and images
with Safranin O. After a careful heating step they have gathered. Proper
to avoid the drying of the stain, the stained acknowledgements were given to the
root tips were then cut so that only the more images as well as references to the
stained portions are retained, discarding drawing.
the less stained and longer ones before Table 2. Data Table showing the Different
transferring them to glass slides. Stages of Mitosis in Onion Root Tip
Coverslips were placed over the cut Karyokinesis
root tips, and to ensure that there were no Interphase
Prophase Metaphase
bubbles, the researchers tapped the
coverslip with their fingers to get rid of the Observed
bubbles. This also allowed for proper (pictures)
squashing of the meristematic tissue,
creating a finer cell layer aiding the
microscope observation. Lastly, the onion
Drawing
tip cells were placed under a microscope;
with label
adjusting it for better focus to obtain clearer cell parts
and more distinct images, as well as using
the scanner (4x), low (10x), and high (40x) The cell
Chromosomes
Chromosomes
prepares itself align at center
objective lenses. for cell
start to
of the cell,
duplicate and
division. It called
Description grows,
nuclear
metaphase
RESULTS AND DISCUSSION develops, and
envelope
plate. And
begins to
The results were gathered during replicates its
shatter
spindles are
chromosome. fully developed
and after the observation of the onion root
tip under the microscope. It was a mixed
data collection because of the utilization of Karyokinesis
other related studies and experiment with Cytokinesis
Anaphase Telophase
the same focus. This is to help in backing
up the results gathered and to determine if Observed
there were concepts missed in the actual (pictures)
activity that should be presented and
observed. The data gathered in the
experiment were the following: structure of
Drawing
each phase and stage of the onion root tip
with label
cell, organelles visible during each phase, cell parts
the magnification of the microscope, and
illustration of the cell through pictures and Sister
Chromosomes The division of
chromatids
drawings. begin to
are contained the parent
in the new cell’s
However, in the actual experiment, Description separate and
nucleus of cytoplasm into
move to
the cell process wasn’t observed clearly opposite poles
each forming two daughter
daughter cell cells occur.
under the microscope due to various of the cell.
reasons particularly the instrument used,
and the way how the onion root tip was Table 2 shows the summary of the
squashed. Due to these limitations, the results and observations gathered during
results of the observation were not the the experiment. At first, it was difficult to
expected results prior to the experiment. adjust the microscope to find a clear image
Thus, other related studies were used to of the cells. Several trials on creating a
proper specimen were done. However, it is CONCLUSION
in the instrument used that had an error. Following the laboratory
The magnification was limited only up to experiment’s objectives, the researchers
x40, that is why the cells were not zoomed were able to draw the following
in to see the actual cell division. Because conclusions:
of this, references from other studies and a. Stages of Mitosis
references were cited to use as guides in • There are three main stages of
creating the results. Mitotic Cell Division: Interphase,
In the table, there are three main Karyokinesis, and Cytokinesis.
stages of mitosis cell division: Interphase, Under Karyokinesis, there are
Karyokinesis, and Cytokinesis. In the four phases such as Prophase,
interphase and Cytokinesis images, the Metaphase, Anaphase, and
documentation of Selene Alegre’s Group Telophase
was in the table to show the difference b. Description of Each Stage
between the before and after of mitosis cell • Interphase – In this stage, cell
division. The reference used in drawing the grows and duplicates its
stages of mitosis was the Image by chromosomes in order to get
Edmund B. Wilson (1900) who observed ready to divide.
onion cells in different stages of mitosis. • Prophase – Thread-like
The same specimen was used by the structures called chromatin
current experiment and Wilson. within the nucleus starts to
Interphase, as seen in the image condense and thicken and
and drawing, is the phase where onion root becomes compacted, resulting
tip cell prepares itself for cell division by in the formation of visible
growing and developing itself and chromosomes. This makes it
replicating its chromosomes. The first visible to a compound light
phase of Karyokinesis begins with microscope. Nuclear envelope
Prophase where the chromosomes of the also starts to shatter.
onion cells are already duplicated the • Metaphase – Chromosomes
nuclear envelope starts to disintegrate. align at the center of the cell
This is also the phase where polar spindles (metaphase plate) and spindles
begin to form. The second phase is the are now fully developed.
metaphase where all the chromosomes • Anaphase – The sister
align at the center of the cell, also known chromatids of the chromosomes
as metaphase plate, and the polar spindles begin to separate and move
are now fully developed. The third phase is towards the spindles in the
Anaphase wherein sister chromatids opposite poles.
separate and begin to move to opposite • Telophase – Both daughter cells
poles of the cell using the polar spindles. have the same number of
For the last phase under Karyokinesis, chromosomes. New membrane
chromosomes are now contained in the for each nucleus also forms from
new nucleus of each forming daughter cell. the pieces of the broken nuclear
And for the last stage of cell division, membrane from the parent cell.
Cytokinesis takes place which is the • Cytokinesis – Cleavage
division of the parent cell’s cytoplasm into increases as this process
two daughter cells to finish the process. continues and the parent cell’s
The mitosis will always start with one cytoplasm divides into two
parent cell and end with two daughter cells.
 genetically identical daughter Wilson, EB. (1900). The cell in
cells. Development and Inheritance. The
Macmilllan Company. 2nd Edition
RECOMMENDATION
With the various limitations APPENDICES
experienced during the conduct of the
laboratory experiment, it is recommended
to do the following to ensure a more
accurate and precise results:
a) Prepare the specimen (onion root
tip) five or seven days before using
it in the experiment;
b) Properly squash the onion root tip
in the glass slides and cover slips to
ensure that a fine layer of the cells
is visible under a microscope; and
c) Use other types of laboratory Figure 1 Growing of Onion Figure 2 Final Set-up of
microscope with higher Root Tip the Specimen Preparation
magnification to make sure that an
image of the specimen can be
zoomed in.

REFERENCES
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Root Tip Cells - Cell Division
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https://byjus.com/biology/study-of-
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Cell cycle. (n.d.). Genome.gov.
https://www.genome.gov/genetics-
glossary/Cell-
Figure 3 Washing with Figure 4 Placing the Roots
Cycle#:~:text=A%20cell%20cycle Distilled Water in a Vial
%20is%20a,mitosis%2C%20and%
20completes%20its%20division
J. Richard McIntosh (2016). Mitosis. doi:
10.1101/cshperspect.023218
John, B. (1990). Meiosis. (No Title).
Murray, A. W. (1994). Cell cycle
checkpoints. Current Opinion in
Cell Biology, 6(6), 872–876.
https://doi.org/10.1016/0955-
0674(94)90059-0.
Schafer KA. The Cell Cycle: A Review.
Veterinary Pathology.
1998;35(6):461-478.
Figure 5 Pouring of Aceto- Figure 6 Heating the
doi:10.1177/030098589803500601 Alcohol Specimen with Alcohol
Burner for 10 seconds
Figure 19 Illustration of Figure 20 Illustration of
Telophase with label of Cytokinesis with label of
Parts Parts

Figure 21 Edmund B. Wilson’s Observation in

Onion Root Tip under a Microscope

Figure 22 Group of Selene Figure 23 Group of Selene

Alegre’s Observation in Alegre’s Observation in
Onion Root Tip under a Onion Root Tip under a
Microscope showing Microscope showing
Interphase Cytokinesis
Figure 7 Heating the Figure 8 Transferring the Figure 13 Image of Onion Figure 14 Zoomed-in
Specimen with Alcohol Roots to Remove Excess Root Tip under the Image of Onion Root Tip
Burner for 10 seconds Water Microscope under the Microscope

Figure 9 Pouring of Figure 10 Squashing the Figure 15 Illustration of Figure 16 Illustration of

Safranin to Stain the Specimen to Make a Fine Interphase with label of Prophase with label of
Sample Layer of Cells Parts Parts

Figure 17 Illustration of Figure 18 Illustration of

Metaphase with label of Anaphase with label of
Parts Parts
Figure 11 Squashed Figure 12 Observing the
Onion Root Tip in a Glass Specimen under a
Slide Microscope