Lab 3 Mitosis and Meiosis

Michelle Seywald

AP Biology

Period 1

January 1, 2010

Introduction:

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 Cells divide by two different methods: meiosis and mitosis. Mitosis is
generally used for the growth and repair of an organism, while meiosis
functions in the production of gametes or spores. Each process involves the
division of the cell’s nucleus and of the cytoplasm.

Mitosis is studied in exercise 3A in cells of whitefish blastula and onion

root tip, since these cells have a high percentage of cells undergoing mitosis.
On roots, the region that contains the highest percentage of cells undergoing
mitosis is called the apical meristem. Cells in their non dividing stage are in
interphase and remain in this stage the majority of the time. DNA is
replicated during this period. At this stage, the nucleus may have one or
more dark stained nucleoli and is filled with chromatin. Cell division begins
with prophase, during which the chromatin condense into chromosomes, the
nuclear envelope disintegrates, and the mitotic spindle begins to form. Next
is metaphase, during which the chromosomes line up at the metaphase plat
in the middle of the cell. In anaphase, the mitotic spindle pulls the sister
chromatids to opposite poles, distributing a diploid amount of chromosomes
to each pole. Telophase, the last stage of division, is characterized by
formation of the nuclear envelope and nucleoli, and uncoiling of the
chromosomes. Following mitosis is cytokinesis, which is division of the
cytoplasm into two daughter cells. The overall process of mitosis and
cytokinesis results in two daughter cells identical to each other and the
parent cell.

Meiosis is a process similar to mitosis in which the nucleus undergoes

two divisions and results in 4 haploid gametes. These two rounds of cell
division are called meiosis I and meiosis II. Meiosis I depends on the
replication of DNA in interphase, just like in mitosis. Beginning with prophase
I, homologous chromosomes come together to form a tetrad and crossing
over takes place between the nonsister chromatids. This process increases
genetic variation in the offspring. At metaphase I, the homologous
chromosomes line up at the metaphase plate. During anaphase I, the
homologous chromosomes are pulled to opposite poles by the spindle fibers.
In telophase I, a haploid set of chromosomes reaches each pole. Cytokinesis
usually occurs during this time. Meiosis II is much like mitosis, with
chromosomes lining up at the metaphase plate and sister chromatids
separating to opposite poles. At the end of telophase II, there are four
haploid daughter cells.

Meiosis and the process of crossing over are explored in exercise 3B

with the fungus Sordaria fimicola. These organisms produce a fruiting body
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called a perithecium containing many asci. These asci each contain 8
ascospores which are either tan or black. The color and orientation of the
ascospores within the ascus can signify whether crossing over has taken
place. If the ascospores are arranged in a 4:4 pattern, crossing over has not
taken place. However, if the ascospores are arranged in either a 2:4:2 or a
2:2:2:2 pattern, crossing over has taken place. By calculating the frequency
of crossover asci within a certain region, the map units between the gene for
spore color and the centromere can be determined.

Materials:

3A.1

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 This exercise requires prepared slides of whitefish blastula and onion
root tip, and a light microscope.

3A.2

This exercise requires a prepared slide of onion root tip and a light
microscope.

3B.2

This exercise requires 1or more slides/images of hybrid asci of the

fungus Sordaria fimicola.

Procedures:

3A.1

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 Examine prepared slides of either onion root tips or whitefish blastula.
Locate the meristematic region of the onion, or locate the blastula, with the
10x objective and then use the 40x objective to study individual cells.
Identify one cell that clearly represents each phase. Sketch and label the cell
in the boxes provided.

3A.2

To estimate the relative length of time that a cell spends in the various
stages of cell division, examine the meristematic region of a prepared slide
of the onion root tip. The length of the cell cycle is approximately 24 hours
for the cells in actively dividing onion root tips. It is hard to imagine that one
can estimate how much time a cell spends in each phase of cell division from
a slide of dead cells, yet this is precisely what will be done in this part of the
lab. Since work will be done with a prepared slide, information about how
long it takes a cell to divide can not be obtained. Instead, the numbers of
cells in each phase will be determined. From this, the percentage of time
each cell spends in each phase can be inferred. Observe every cell in one
high-power field of view and determine which phase of the cell cycle it is in.
This is best done in pairs. The partner observing the slide calls out the phase
of each cell while the other partner records. Then switch so the recorder
becomes the observer and vice versa. Count at least two full fields of view. If
less than 200 cells have been counted, then count a third field of view.
Record the data. Calculate the percentage of cells in each phase and record.
Consider that it takes, on average, 24 hours (or 1,440 minutes) for onion root
tip cells to complete the cell cycle. The amount of time spent in each phase
of the cell cycle can be calculated from the percentage of cells in that stage.

3B.2

View the slide and locate a group of hybrid asci (those containing both
tan and black ascospores). Count at least 50 hybrid asci and enter the data
in Table 3.3. Using the data in Table 3.3, determine the distance between the
gene for spore color and the centromere. Calculate the percentage of
crossovers by dividing the number of crossover asci (2:2:2:2 or 2:4:2) by the
total number of asci x100. To calculate the map distance, divide the
percentage of crossover asci by 2. The percentage of crossover asci is
divided by 2 because only half of the spores in each ascus are the result of a
crossover event. Record the results in Table 3.3.

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Results:

3A.1

Sketches attached.

Analysis Questions:

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1. Explain how mitosis leads to two daughter cells, each of
which is diploid and genetically identical to the original cell.
What activities are going on in the cell during interphase?

First, mitosis replicates the chromosomes in a cell, ensuring that

there are two identical copies of DNA. At metaphase, the
replicated chromosomes line up at the metaphase plate. At
anaphase, the sister chromatids, which are identical to each
other, are pulled apart to opposite poles by the spindle fibers.
The elongated cell then separates into two diploid cells, each
with genetic material identical to each other and to the parent
cell. The daughter cells remain diploid because homologous
chromosomes do not separate as they do in meiosis, but rather
remain together as a diploid set. The genetic material is identical
in the parent and the daughter cells because anaphase
separates replicated chromosomes and distributes identical
copies of chromatids to each daughter cell. In addition, crossing
over does not take place during mitosis, preventing sister
chromatids from differing from each other.

2. How does mitosis differ in plant and animal cells? How does
plant mitosis accommodate a rigid inflexible cell wall?

While animal cells use centrioles to direct the spindle fibers,

plant cells lack these structures. This, however, does not seem to
significantly affect the function of the microtubules of the spindle
in plant cells. Another difference is the formation of a cell plate
during cytokinesis in plant cells that is not present in animal
cells. Rather than pinching off along a cleavage furrow to form
two new daughter cells, a new cell wall is simply laid down
between the daughter cells in plant cells.

3. What is the role of the centrosome (the area surrounding

the centrioles)? Is it necessary for mitosis? Defend your
answer.

The centrosome contains a pair of centrioles and functions as the

microtubule organizing center. If this structure were absent from
the cell, the microtubules would not assemble, the spindle would
not form, and the sister chromatids would not separate. In short,
mitosis would not be possible without the presence of the
centrosome.
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3A.2

Table 3.1

Number of Cells Percent Time in

of Total Each
Field 1 Field 2 Field 3 Total Cells Stage
Counted
Interphas 946.08
70 64 63 197 65.7%
e minutes
240.48
Prophase 15 18 17 50 16.7%
minutes
Metaphas 95.04
5 8 7 20 6.6%
e minutes
72
Anaphase 4 4 7 15 5%
minutes
Telophas 86.4
6 6 6 18 6%
e minutes
Total Cells Counted 300

Questions:

1. If your observations had not been restricted to the area of

the root tip that is actively dividing, how would your results
have been different?

If the observations had not been confined to the actively

dividing portion of the onion root tip, there would be virtually
no cells in prophase, metaphase, anaphase, and telophase.
Instead, most cells would be in interphase.

2. Based on the data in Table 3.1, what can you infer about the
relative length of time and onion root tip cell spends in each
stage of cell division?

Based on the data in Table 3.1, it can be inferred that an

onion root tip cell spends the majority of its time in
interphase, followed by prophase, then equal amounts in
metaphase and telophase, and finally the least amount in
anaphase.

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 3. Draw and label a pie chart of the onion root tip cell cycle
using the data from Table 3.1

3B.1

1. List three major differences between the events of mitosis

and meiosis.

One major difference between mitosis and meiosis is that

crossing over takes place during meiosis but not during mitosis.
Also, mitosis produces 2 identical daughter cells, while meiosis
produces 4 potentially unique daughter cells. Finally, the nucleus
undergoes two divisions in meiosis, but only one division in
mitosis.

2. Compare mitosis and meiosis with respect to each of the

following in Table 3.2:

Mitosis Meiosis
Chromosome Number
Diploid (2n) Diploid (2n)
of Parent Cells
Number of DNA
1 1
Replications
Number of Divisions 1 2
Number of Daughter
2 4
Cells Produced
Chromosome Number
Diploid (2n) Haploid (n)
of Daughter Cells
Production of
Purpose/Function Growth and repair
gametes

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 3. How are meiosis I and meiosis II different?

Meiosis I undergoes replication of DNA and separates

homologous chromosomes into daughter cells, while meiosis II
does not replicate DNA before division and separates sister
chromatids into daughter cells.

4. How do oogenesis and spermatogenesis differ?

Oogenesis results in only one viable egg cell, while

spermatogenesis results in 4 functioning sperm cells.

5. Why is meiosis important for sexual reproduction?

Meiosis halves the chromosome number of a cell so that when

two gametes fuse together and each contributes their set of
chromosomes, the diploid number of chromosomes is preserved.
Meiosis also involves crossing over, which increases the genetic
variation of the offspring.

3B.2

Table 3.3

Number of % Asci Gene

Number of
4:4 Showing Centromere
Asci Showing Total Asci
Crossover distance
Crossover
Divided by 2 (map units)
30 51 81 31.4% 31.4

Analysis of Results:

2.

Meiosis 1
Meiosis 2

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Data Analysis:

3A.1:

This experiment focuses on observing the cells of whitefish

blastula and onion root tip under a light microscope. Had the
microscope light been faulty or weak, the student would not be able to
differentiate between the different stages of cell division or the specific
structures in each stage. Also, those slides more heavily dyed
produced a clearer image of certain structures that groups with lighter
dyed slides were unable to see. These possibilities may have
contributed to the inaccurate calculation for the duration of telophase.
Although it is the shortest stage of mitosis, it was found to the second
to last shortest in this lab.

3A.2:

This experiment concentrates on determining the percentage of

time spent in each stage of the cell cycle by observing a large number
of cells and recording their particular stage. This experiment requires
the student to count a large number of cells in the meristematic region
of the onion root tip in order to determine the percentage of time spent
in each stage of cell division. If the student was unable to differentiate
between certain similar stages, such as prophase and interphase, the
data may have been miscalculated. In addition, the percentage of cells
found in each stage may vary from slide to slide, or from region to
region observed under the microscope.

3B.2:
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 This exercise focuses on calculating the map units between a
specific gene and the centromere by observing the frequency of
crossover asci from the fungus Sordaria fimicola. There was limited
room for error, since the images of the hybrid asci were already
prepared and the student needed only to tally the number of 4:4 and
crossover asci. However, if there was difficulty differentiating between
crossovers and non crossovers, or certain asci were counted more than
once, the results may have been skewed.

Conclusion:

Mitosis and meiosis were observed throughout this lab under

microscope and through simulation which helped clarify their many
similarities and differences. In exercise 3A, the different stages of cell
division of whitefish blastula and onion root tip were observed and
recorded. The frequency of each stage in a region of cells was then
calculated and used to determine the amount of time a cell spent in
each stage of cell division. From this data, it can be concluded that
prophase is the longest stage of mitosis, and telophase is the shortest.
In Lab 3B, meiosis was simulated and examined the link between
crossing over and map units. It can be seen from observation of
mitosis and meiosis that, although similar, the two processes have
many differences. For one, the cell nucleus undergoes two divisions in
meiosis, but only one division in mitosis. Secondly, crossing over takes
place during meiosis but not during mitosis, increasing the genetic
variation of offspring. In addition, mitosis produces 2 identical daughter
cells, while meiosis produces 4 potentially unique daughter cells. In
exercise 3B.2, the frequency of crossover asci (2:4:2 or 2:2:2:2
formation) from the organism Sordaria fimicola is calculated and used
to determine the approximate distance from the gene for spore color
to the centromere. From the results, it can be concluded that the gene
is approximately 31.4 map units from the centromere.

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