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Mitosis and Meiosis

Part I Mitosis

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It was discovered in 1858, by Rudolf Virchow, that new cells can only arise from previously
existing cells. This is done in two ways: mitosis and meiosis. Somatic (body) cells divide
exclusively by mitosis followed by cytokinesis, while germ cells produce gametes by the process
of meiosis. Plant cells grow by enlargement, essentially by taking up water. When they reach a
certain size, they divide, forming two identical daughter cells. The various parts of the cell are
divided in such a way that the new daughter cell is identical to the parent cell.

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Strictly speaking, mitosis implies only the division of the nucleus, and is therefore distinct from
cell division, in which the cytoplasm is divided. In most organisms, cells divide by ingrowth of
the cell wall, if present, and the contraction of the cell membrane, a process that cuts through the
spindle fibers. In land plants (bryophytes and vascular plants) and a few algae, cell division takes
place by the formation of a cell plate. Small droplets appear across the equatorial plate of the cell
and gradually fuse, forming a disc that grows outward until it reaches the wall of the dividing
cell, which completes the separation of the two daughter cells.

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The DNA of prokaryotes is simply replicated before division. In eukaryotes, however, the
hereditary material is part of their complex chromosomes. Equal division of this material requires
a more complex method by which the chromosomes are replicated, separated, and apportioned
precisely between the daughter cells.
Mitosis, or nuclear division, ensures the equal division of the nuclear material between the
daughter cells in eukaryotic organisms. During mitosis the chromosomes condense, and move to
the center of the cell where they fully contract. They then split longitudinally into two identical
halves that appear to be pulled to opposite poles of the cell by a series of
microtubules. In these two genetically identical groups, the coiling of the
chromosomes relaxes again, and they are reconstituted into the nuclei of the
two daughter cells. It is a continuous process that can be divided into five
major phases: interphase, prophase, metaphase, anaphase, and telophase.

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Interphase: The chromatin, if visible at all, can only be seen as small

grains or threads. Interphase is generally considered to be a resting phase.
However, the cell is replicating the genetic material, preparing for mitosis.

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Prophase: The beginning of mitosis is illustrated by the chromosomes

gradually becoming visible. They start out as elongated threads that shorten
and thicken. Chromosomes become more condensed and undergo spiral
contractions, like a thin wire being turned into a coiled spring. This coiling
involves the entire DNAprotein complex. Each chromosome is composed
of two longitudinal halves, called chromatids, joined in a narrow area
known as the centromere, where the chromatids are not coiled. The
centromere, located on each chromosome, divides the chromosomes into
two arms of varying lengths. As prophase progresses, the nucleoli grow
smaller and finally disappear. Shortly after, in most cell types, the nuclear
envelope breaks down, putting the contracted chromosomes into direct
contact with the cytoplasm; this marks the end of prophase.

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Metaphase: The chromosomes, still doubled, become arranged so that each
centromere is on the equatorial region of the spindle. Each chromosome is
attracted to the spindle fibers by its centromere; often the arms of the
chromosome point toward one of the two poles. Some of the spindle fibers
pass from one pole to the other and have no chromosome attached.

Anaphase: The chromatids separate from one another and become

individual chromosomes. First, the centromere divides and the two daughter
chromosomes move away from the equator toward opposite poles. Their
centromeres, which are still attached to the spindle fiber, move first, and the
arms drag behind. The two daughter chromosomes pull apart; the tips of the
longer arms separate last. The spindle fibers attached to the chromosomes
shorten as the chromatids divide and the chromosomes separate. The fibers
appear to move, but in fact the microtubules are continuously formed at one
end of the spindle fiber and disassembled at the other. In the process, it
appears as if the spindle fibers were tugging the chromosomes toward the
poles by their centromeres. By the end of anaphase, the two identical sets of
chromosomes have separated and moved to opposite poles.
Telophase: The separation is made final; the nuclear envelopes are
organized around the two identical sets of chromosomes. The spindle
apparatus disappears. Nucleoli also reform at this time. The
chromosomes become increasingly indistinct, uncoiling to become
slender threads again.

Cytokinesis: As mitosis ends, cytokinesis begins, resulting in the

formation of two daughter cells. The cleaved membrane slowly draws
together, forming a narrow bridge, then separates the cell into two
daughter cells. The cells now enter interphase.

Mitosis ends when the processes are complete and the chromosomes
have once more disappeared from view. The two daughter cells enter
interphase. The two daughter nuclei produced are identical to one
another and to the nucleus that divided to produce them.
In order to investigate the process of mitosis, plant and animal tissues where cells are dividing
rapidly must be examined. In animals, the most rapidly growing and dividing tissues are found in
the embryonic stages of development. Although most animal tissues continue to undergo mitosis
throughout the life cycle of the organism, they do so very slowly when compared to their
embryos. Some animal cells, like most plant tissues, rarely replicate after the organism reaches
maturity.
In plants, these tissues are primarily found in the tips of stems and roots. The root tip plants are
exceptionally good places to look for cells undergoing mitosis. Plant root tips consist of several
different zones where various developmental and functional processes of the root are performed.
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Mitosis and Meiosis

The primary region for the formation of new cells is the apical meristem. The root cap offers
protection for the rest of the root, the region of elongation is the area where the bulk of cell
growth occurs, and the region of maturation is where tissue differentiation occurs.

Part II Meiosis
Sexual reproduction provides a mechanism to produce genetic variation, as the genes of two
different individuals are arranged in various ways. This requires a reduction in the chromosome
number of the parent cell, normally diploid, to half that, or haploid, in somatic cells. The type of
cell division resulting in half the chromosome number of the parent cell is called meiosis.
In meiosis, a germ cell divides into four
haploid gametes. When two gametes
egg and spermcombine during
fertilization, forming a zygote, the diploid
chromosome number is restored. Meiosis
consists of one DNA replication and two
nuclear divisions, meiosis I and II. This
results in the formation of four daughter
cells, each with only half the number of
chromosomes as the parent.
Genetic variability is further increased by
a process called crossing over. In the early
stages of meiosis, the homologous pairs
of chromosomes move close together in
such a way that all four chromatids are
entwined, forming a tetrad. This process,
known as synapsis, allows for the
exchange of chromosome sections
between the homologous pairs.
The example that will be used in the
investigation is Sordaria fimicola, an
ascomycete fungus that is haploid for the
bulk of its life cycle, including the
individual fungal filaments, called
hyphae, which normally exist in a mass
called a mycelium representing the
body of the fungus; and the ascospores,
from which mycelia develop. The only
diploid portion of the life cycle of S. fimicola occurs when the nuclei of specialized hyphae come
together.
These hyphae, which belong to different strains of the species, fuse to form a zygote. This zygote
then undergoes meiosis to produce the haploid ascospores, yielding four haploid nuclei contained
in a sac called an ascus. After meiosis, the four nuclei undergo mitosis, resulting in an ascus
containing eight haploid ascospores. Many asci form inside a fruiting body called a perithecium.

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One type of genetic variability in S. fimicola is the color of the ascospores. Most strains are the
dark brown, wild-type ascospores, although there are variants. Certain strains have tan or gray
ascospores. A tan ascospore strain mated with the wild-type variety produces a series of
perithecia containing asci with four tan and four wild-type ascospores each.
How these ascospores are arranged within the ascus is a direct representation of whether or not
crossing over has occurred between the centromere and the site for the gene for ascospore color.
If no crossing over has occurred, the ascospores will be arranged in a 4:4 manner. If crossing
over has occurred, they will occur in a 2:4:2 or 2:2:2:2 manner.
By observing the ascospore arrangement, the percentage of asci exhibiting crossover can be
determined. This frequency appears to be affected largely by the distance from the gene to the
centromere. From the crossover frequency, the distance in map units from the gene for ascospore
color, and the chromosome centromere, can be calculated.

OBJECTIVES
In this experiment, you will
Examine and compare the phases of mitosis in animal and plants cells.
Determine the relative time cells spend in each phase of mitosis.
Prepare microscope slides of mitotic cells using onion Allium root tips.
Follow the processes of mitosis and meiosis in the life cycle of Sordaria.
Examine the arrangement of Sordaria ascospore microscopically to determine the
frequency of crossing over.
Calculate the distance, in map units, between a specific gene and the chromosome
centromere.

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Mitosis and Meiosis

Part I Mitosis
MATERIALS
Materials for Parts A and B

whitefish mitosis slide

onion mitosis slide
compound microscope
Materials for Part C

onion root tip

hydrochloric acid 1M
toluidine blue 0.5%
compound microscope
coverslip
microscope slide

Bunsen burner
clothespin or forceps
paper towel
pipet
scalpel

PROCEDURE
Part A: Observing Mitosis in Plant and Animal Cells

1. Observe the prepared microscope slide of onion root tip mitosis, first at 100X, then 400X.
Using the Plant Mitosis Chart as a guide, identify cells that represent each mitotic phase.
2. In the Analysis section, draw each phase of plant cell mitosis that you see. Write a brief
description of each phase below each drawing.
3. Observe the prepared microscope slide of whitefish blastula. Using the Animal Mitosis Chart
as a guide, identify each phase of animal cell mitosis.
4. In the Analysis section, draw each phase of plant or animal cell mitosis that you see. Write a
brief description of each phase below each drawing.
Part B: Relative Lengths of Phases of Mitosis

5. Examine at least three fields of view of the apical meristem of the onion root tip at 400X. In
each view, count the number of cells in the various stages of mitosis. Record this data in
Table 1.
6. Calculate the total number of cells counted and the percentage of total cells counted for each
stage of mitosis. Record this data in Table 1. Record the percentages in Table 2, as well.
7. Assuming that it takes an average of 24 hours (1,440 minutes) for onion root tip cells to
complete the cell cycle, calculate the amount of time cells spent in each phase of the cycle.
Use the formula provided below. Enter your results in Table 1.
Percent of Cells in Phase 1,440 minutes = _________ minutes cell spent in phase

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Part C: Preparation of an Onion Root Tip Squash

8. Wrap a fresh onion set with paper toweling and place it in a plastic sandwich bag. Add 10 mL
of water before sealing the bag.
Note: The set used must have root primordia present or it will not produce root tips. The root
tips will grow within 36 to 72 hours. Common onions have a mitotic cycle of approximately
20 hours.
9. Place the plastic bag in a box or dark place until the root tips have grown to a length of about
4 to 5 mm. It is important that the onion grows in the dark to ensure that it produces roots
rather than shoots.
10. Remove the onion from the box approximately one half-hour before performing the
experiment to expose the root tips to light.
11. Blot as much excess water from the root tips as possible. Any excess water on the slide will
affect your results. Do not allow the root tips to dry out.
12. Using a scalpel or razor blade, cut off the end of one of the emergent root tips; the section
should be approximately 1 to 2 mm long. Place the root tip on a clean microscope slide and
apply one to two drops of HCl on the root tip.
13. Holding the slide with a clothespin or forceps, pass it through the flame of a Bunsen burner
for several seconds. Note: Do not hold the slide directly over the flame.
14. Without harming the root tip, blot the specimen with a paper towel to remove the excess HCl.
Note: You may wish to touch a corner of the paper towel to the edge of the root tip and allow
the paper towel to wick up the solution.
15. Add one to two drops of 0.5% aqueous toluidine blue stain to the root tip. Note: Toluidine
blue is a mild irritant, avoid direct physical contact with this stain.
16. Pass the slide through the flame of a Bunsen burner for one to two minutes. Let the slide
stand and cool for one minute. Note: Do not hold the slide directly over the flame.
17. Remove excess stain with a paper towel in a motion similar to the one used in Step 14.
18. With a scalpel or razor blade, slice the root tip longitudinally down the middle into two
mirror segments.
19. Add a drop of toluidine blue and cover the root tip with a coverslip. Using the scalpel handle
or other blunt instrument, gently press down on the coverslip to squash and spread out the
root tip. Blot off excess stain, if any, that may come out from under the coverslip.
20. View the slide under a microscope using the 10X objective. Locate the apical meristem.
Examine the slide using the 40X objective. Locate cells in the various stages of mitosis.
Sketch the nuclear regions of individual cells in the five different stages and describe key
features of each in the Analysis section. Keep in mind that since the root tip has been
squashed, the meristem may not be readily recognizable.

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Mitosis and Meiosis

Part II Meiosis
MATERIALS
microscope slide
coverslip
inoculating loop
sordaria demonstration cross plate (shared)

PROCEDURE
1. Place a drop of water on a clean slide with an inoculating loop.
2. With an inoculating loop, scrape several perithecia from the
demonstration cross plate. Scrape the perithecia from the interface of
two crossing strains close to the edge of the plate and place in the
drop of water on the slide. Avoid picking up agar along with
perithecia; it will interfere with results.
3. Cover the slide with a coverslip. Using a pencil eraser or other blunt
instrument, gently press down on the coverslip to squash and spread
out the perithecia. The pressure should be sufficient to squeeze the
asci from the perithecia, but not enough to crush the asci themselves. Note: It may be helpful
to slide the coverslip around on top of the sample, with slight pressure, to spread out the asci
and make them easier to observe. Keep in mind, however, that applying too much pressure
may rupture the asci, releasing the individual ascospores.
4. View the slide under a microscope at 100X. Locate the asci. You may wish to view the slide
at 400X to determine the color of some ascospores. The slide preparation should show
collapsed perithecia and asci clusters (rosettes), with mature ascospores in various
arrangements. Immature ascospores will all be light colored. Since S. fimicola is homothallic,
the preparation will show both hybrid and self-fertilized perithecia of both parental types.
Hybrid perithecia, however, will not occur very far from the line of contact between the two
varieties. Prepare three slides to get an adequate sampling of hybrids, if possible.
5. Count approximately 50 hybrid asci from at least three fields of view, preferably from
different slides. Record the data in Table 2.

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ANALYSIS

Stage: ___________
Description: ______
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Stage: ___________
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Stage: ___________
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Stage: ___________
Description: ______
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Stage: ___________
Description: ______
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Mitosis and Meiosis

Table 1
Cells in each stage of mitosis
Stage

# of cells in
Field 1

# of cells in
Field 2

# of cells in
Field 3

Total # of
cells

% of total
# of cells

Time of each
stage (min)

Interphase
Prophase
Metaphase
Anaphase
Telophase

Total number of cells counted __________

Table 2
Percentage of cells in each stage of mitosis
Stage

% of total # of cells

Interphase
Prophase
Metaphase
Anaphase
Telophase
Total

Table 3
Observed Sordaria Asci
# of 4:4

# of crossover

Total

% asci showing
crossover

Gene distance from

centromere

Using your data, determine the distance in map units from the gene for ascospore color to the
chromosome centromere. Calculate the percentage of asci that showed crossover. This
percentage crossover must be divided by two, since only half the ascospores in each hybrid ascus
are the result of crossing over. Dropping the % symbol gives you the map distance from the gene
to the centromere. Record the data in Table 3.
% Crossover = # showing crossover 100%
total counted
Gene Distance from Centromere = % Crossover
2

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QUESTIONS
1. Referring to the percentage of total cells counted in each phase of mitosis, determine which
phase takes the longest for the cell to complete, and explain why. Sketch a pie graph of the
percentage of cells in each phase to illustrate. Be sure to title your graph and include a key.
2. What is the relationship between the processes of mitosis and cytokinesis?
3. Which of the following is significantly different between plant and animal cell mitosis?
a.
b.
c.
d.

metaphase
anaphase
cytokinesis
prophase

4. How does meiosis lead to genetic variability within a population? Use S. fimicola as an
example.
5. Define the following terms.
somatic cell
germ cell
chromatin
centromere
diploid
haploid
zygote
6. Create a Venn diagram showing at least two similarities and two differences between mitosis
and meiosis.

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Mitosis and Meiosis

ANIMAL CELL MITOSIS

Interphase

Prophase

Metaphase

Anaphase

Telophase

Cytokinesis

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PLANT CELL MITOSIS

Interphase

Prophase

Metaphase

Anaphase

Telophase

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Cytokinesis

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