LAB-2:

VISUALISATION OF DIFFERENT PHASES OF MITOSIS IN

ONION TIP ROOT CELLS
NAME-NISHCHAY CHAUDHARY
ROLL NUMBER-2023UMA0226
AIM:
This experiment will focus on understanding cell division in plant cells. Cell division is
of two types: mitosis and meiosis. Apart from gametes involved in reproduction,
most of the cells undergo mitosis for cell division and tissue growth. In this
experiment using onion root tip cells we will observe and study the different stages of
mitosis and to quantify the frequency of cells undergoing mitosis.

INTRODUCTION:
Mitosis is a fundamental process of cell division found in all eukaryotic cells, which
plays a critical role in tissue growth, repair, and the production of genetically identical
daughter cells. The complex and highly regulated process involves the replication and
separation of chromosomes, as well as the partitioning of cytoplasmic contents. The
mitotic process occurs in several distinct stages: prophase, metaphase, anaphase,
and telophase. During prophase, chromatin condenses into visible chromosomes, and
the nuclear envelope disintegrates. Chromosomes then line up along the equatorial
plane of the cell during metaphase.
Sister chromatids are separated and pulled towards opposite poles of the cell during
anaphase, while in telophase, the chromosomes decondense and the nuclear
envelope reforms around the two sets of chromosomes.
Finally, the cytoplasm is divided through cytokinesis. Mitosis is vital for the proper
functioning of multicellular organisms, and any aberration in the process can result in
developmental abnormalities and diseases such as cancer. Therefore, comprehending
the mechanisms underlying mitosis is crucial for both basic and applied research
purposes.

APPARATUS:
Onion Bulbs. scalpel, microscopic slides and coverslips, glacial acetic acid, ethanol, 1%
HCl solution, Aceto-carmine stain, microscope, distilled water.
PROCEDURE:
Growing of root tips:
● Choose a few medium-sized onion bulbs and remove the dry roots.
● Grow new roots by placing the bulbs on glass tubes (of about 3–4 cm. diameter)
filled with water, ensuring that the stem part just touches the water. Add water drops
as needed to compensate for evaporation. It may take 3-6 days for new roots to grow.
● Once the new roots are 2-3 cm long, cut them and transfer them to a fixative made
of aceto- alcohol (1:3 ratio of glacial acetic acid to ethanol).
● Let the root tips sit in the fixative for 24 hours, then transfer them to 70% ethanol
for preservation and future use.
● Note that onion root-tip cells typically divide once every 24 hours, with the division
occurring about two hours after sunrise. To observe the highest number of dividing
cells, cut the roots grown in water only at that time.
Preparation of slide:
● Take one or two preserved roots and wash them on a clean and grease-free slide
with water.
● Place one drop of HCl followed by 2-3 drops of aceto-carmine stain on the root tip.
● Put the root tips in a test tube with dilute aceto-carmine solution and heat the test
tube in a water bath
at 60°Cfor about 10 minutes.
● Cut the tip portion of the root that is more stained (2-3 mm), and discard the
remaining portion.
● Add a drop of water on the root tip and place a coverslip on it, avoiding air bubbles.
● Hold the slide in between the folds of blotting paper using your fingers to ensure
the cover slip is properly held.
● Use the blunt end of a pencil to gently tap the coverslip, so that the meristematic
tissue of the root tip below the coverslip is properly squashed and spread into a thin
layer of cells.
● Carefully seal the margins of the coverslip using molten paraffin wax or nail polish.
● The onion root tip cells preparation is now ready for the study of mitosis .
OBSERVATION:

metaphase

Late anaphase

anaphase

prophase
INFERENCE:
The experiment conducted on onion root tip cells aimed to deepen our
understanding of cell division processes, specifically focusing on mitosis. Mitosis is a
fundamental mechanism for cell division and tissue growth in most cells, excluding
gametes involved in reproduction. Through the examination of onion root tip cells,
we were able to observe and analyse various stages of mitosis.
The microscopic examination revealed distinct phases of mitosis, including prophase,
metaphase, anaphase, and telophase. Each stage displayed characteristic features,
such as chromosomal condensation and alignment, chromatid separation, and the
formation of daughter cells. The observed patterns provided valuable insights into
the dynamic and precisely regulated nature of mitotic cell division.
Interphase is the most significant phase of the cell cycle for several reasons. One
explanation for this is that the cell readies itself for cell division during interphase.
During interphase, the cell grows and replicates as well. Cell division occurs during
theG1, G2, and S stages.
In conclusion, the experiment using onion root tip cells enhanced our comprehension
of mitosis, shedding light on its intricacies and highlighting its significance in cellular
proliferation and tissue growth. The obtained data not only facilitated the
identification of mitotic stages but also enabled the quantification of mitotic activity.
This knowledge contributes to the broader understanding of cell biology and provides
a foundation for future research in the field.

TAKE HOME QUESTIONS:

1. Why mitosis is also called as the equational division?
Ans. Mitosis is called "equational division" because it results in two daughter cells
that are genetically identical to the parent cell and to each other. The chromosome
number remains the same, ensuring the preservation of genetic information for
growth and tissue maintenance.
2. What are the consequences of errors in mitosis, and how can such errors
contribute to disease?
Ans. Errors in mitosis can lead to chromosomal abnormalities, causing various health
issues. These errors, such as nondisjunction or chromosome breakage, can result in
abnormal cell division and contribute to conditions like cancer, genetic disorders, and
developmental abnormalities. Certainly. Two examples of diseases associated with
errors in mitosis are:
Down Syndrome (Trisomy 21): Down syndrome is caused by an error in mitosis
known as nondisjunction, where chromosomes fail to separate properly during cell
division. In the case of Down syndrome, there is an extra copy of chromosome 21,
leading to developmental and intellectual delays, characteristic facial features, and an
increased risk of certain health issues.
Cancer: Many cancers are linked to errors in mitosis that result in uncontrolled cell
growth. Mutations in genes that regulate the cell cycle can lead to the formation of
tumours. For example, mutations in tumour suppressor genes or oncogenes can
disrupt the normal progression of mitosis, allowing cells to divide uncontrollably,
forming tumour’s that may become cancerous.
4. How do different cell types vary in terms of their rates of mitosis, and what factors
determine these differences?
Ans. Different cell types vary in their rates of mitosis based on tissue function,
developmental stage, and external stimuli. Tissues with high turnover, like skin or
digestive lining, undergo frequent mitosis for repair. Specialized cells, such as
neurons, have limited mitotic activity. Developmental stages also influence mitotic
rates, with embryonic cells dividing rapidly for growth. External factors like wound
healing or hormonal influence can trigger increased mitosis. Additionally, intrinsic
genetic factors play a role, determining whether a cell is programmed for frequent
division or not. Understanding these variations are essential for grasping tissue
dynamics and overall organism maintenance.
5. How might new imaging technologies, such as super-resolution microscopy, help
us to better understand the mechanics of mitosis?
Ans. Super-resolution microscopy enhances our understanding of mitosis by
providing higher resolution and clearer images, enabling the observation of finer
cellular structures and dynamic processes in real-time. Its three- dimensional imaging
capabilities offer depth perception, while the ability to track individual molecules
allows for a detailed analysis of mitotic components. The precision of measurements
and integration with other imaging modalities contribute to a comprehensive view of
mitotic mechanics. This advanced technology revolutionizes our ability to study and
uncover details previously inaccessible with traditional microscopy.
6. Where does the spindle fibre originate in the cell?
Ans. Spindle fibres, or microtubules, originate from centrosomes in the cell.
Centrosomes, composed of centrioles and pericentriolar material, move to opposite
poles during mitosis, and microtubules extend from them to form the mitotic spindle.