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Max Louis Isabelle

BIO110H LAB
Jesse Weber
10/24/15
The Effects of Exposure to X-Rays on the crossover frequency
of Sordaria Fimicola
Introduction
Background Information:
Sordaria Fimicola is a model organism that is commonly used in
genetic experiments. A model organism is one that has been widely
studied and possesses the ability to breed and survive in a laboratory
setting. These traits supply scientists with the capacity to use Sordaria
Fimciola easily in the laboratory setting for a multitude of different
experiements.

...The products of meiosis (the tetrad of two pairs of sister


chromatids) show up in the ascospores in the exact order in which they
separated from one another(Volk). Ascospores are the result of a
meiotic division followed by a mitotic division, resulting in a line of
eight ascospores in the ascus (a sac in which spores develop). What
this means is that recombination in this organism can be observed

exactly how it occurs in the three different asci types. Out of the three
asci types, two represent recombination (2:4:2 and 2:2:2:2) and one
represent non-recombinant (4:4). The ratios are in respect to the two
colors observed in the ascospores. These ratios are where the result of
the experiment, that the author conducted with his classmates, in Bio
110H at PSU is derived.

Purpose:
The purpose of the experiment the author conducted is to examine the
recombination rates of Sordaria Fimicola and see if the environment in
which the Sordaria developed can influence them. Recombination
generates new combinations of existing genetic variation and therefore
may be important in adaptation and evolution (Saleem, Lamb, Nevo).
This quote explains the purpose of this experiment, which is to
examine the mechanism for adaptation and evolution, by isolating a
single variable to test, in this case the environment.

Hypothesis:
The author hypothesized that the Sordaria Fimicola, which is exposed
to the x-ray treatment, will express a higher frequency of
recombination compared to the control Sordaria Fimicola.

Summary of Procedures:

To test this hypothesis, the author used two agar plates, one was the
control and the other was exposed to the treatment. Each agar plate
was divided into four quadrants. For this experiment, two strains of
Sordaria were used, one wild type the other tan. The strains of the
Sordaria were placed on non-adjacent quadrants on the agar plate.
This process was completed for both plates. The reason for placing the
two strains in non-adjacent quadrants is to ensure the two strains are
in an ideal location to sexually reproduce. The treatment was exposed
to the x-rays for two weeks and the control was left in a stable
environment. After two weeks the two plates were collected and a
squash was performed. A squash is where you use an inoculating loop
and scrape some of the bacteria colony from the agar plate and place
it on a microscope slide. Then a drop of water was placed on the
bacterial colony, followed by a coverslip. After the cover slip is placed
over the slide a gentle pressure was applied to the cover slip. This is
done to try to release the asci from the perithecia without breaking the
ascus. Once this is done correctly, a count was recorded of the ratios
of the asci under a microscope. As previously stated two of the ratios
signify crossing over and one does not. The frequency of each ratio
was recorded and the results were analyzed.(Burpee, Cyr, Hass, Ikis,
Richter, Ward, Woodward).

Material and Methods:

The experiment that the author conducts is designed to test the effects
of the environments (IV) on the recombination frequency (DV) of the
Sordaria. The Control in this experiment is normal sustained
temperature and conditions, while the treatment is non stop exposure
to x-ray radiation for two weeks. To prepare the control and treatment,
the Sordaria was plated on an agarose solution using inoculating loops.
For the collection of the data, inoculating loops were used to place the
Sordaria that had been growing for the past two weeks, in its
respective environments, on the microscope slides. The method used
for the collection of the data is to count the resulting asci types of the
control and treatment, using a microscope, specifically comparing the
frequency of recombination in the control vs. the treatment. The
author compiled his data with twenty-two other groups to create a data
set that is comprised of 460 asci for control and 460 for treatment.
The recombination rates were then found as a frequency. Before the
author converted that frequency to a percent, graphs and tables were
made to easily show the author and the twenty-two other groups
involved data.

Graphs and Figures


Trends

Treatment (Figure 1)
350
300
250
Treamtment

200
150
100
50
0
Non-recombinant

Recombinant

Figure 1:1 shows that in the Sordaria Fimicola exposed to the treatment
there was a significant difference between the frequency of nonrecombinnat and recombinat asci types.

Control (Figure 2)
300
250
200

Control

150
100
50
0
Non-recombinant

Recombinant

Figure 1:2 shows that in the Sordaria Fimicola exposed to the control
environment the frequency of non-recombinant and recombinant asci
types were about the same.

Combined Data (Figure 3)


350
300
250

Treamtment

200

Control

150
100
50
0
Non-recombinant

Recombinant

Figure 2:1 shows that the environment had an affect on the frequency
of the non-recombinant and recombinant asci types in the Sordaria.

Combined Section Treatment and Control (Table 1)


Non
recombina
nt
# Of Type
A Asci
(4:4)

Treatme
nt
Control

Recombinant

Total
#
Of
Asci

Total#
Recombina
nt Asci
(B+C)

Frequency
of
Recombina
nt Asci
(B+C)/total

Frequenc
y of Type
B Asci
(B/total)

Frequenc
y of type
C asci
(C/total)

Ratio
B/C

0.975
5
1.105

# Of
Type C
Asci
(2:2:2:
2)

138

# Of
Type
B
Asci
(2:4:2
)
159

163

460

322

0.766

0.3457

0.354

220

126

114

460

240

0.5217

0.2739

0.2478

Discussion
Interpretation of Data:

When the Figure (1:2) and Figure (1:1) are combined in Figure (2:1) it is
easy to see that the environment (IV) had some affect on the
recombination frequency in the Sordaria (DV). When the Sordaria was
exposed to the treatment environment, it displayed a higher frequency
of recombination. The harsh environment of growing under a constant
barrage of x-ray radiation some how causes the Sordaria to adapt and
recombine, at a higher frequency, to create genetic diversity. Under
normal conditions the Sordaria level of recombination is about 52%
while under x-ray it is 76.6%. The reason this recombination rate is so
high in the Sordaria exposed to x-rays, could be similar to how x-rays
can cause cancer. ...x-rays and atomic particles not only modify DNA,
but also can cause cancer in animals and can transform normal cells in
culture into rapidly proliferating, cancer-type cells (Freeman). Since
cancer affects cell division and x-rays tend to increase cancer rates in
cells, it would be expected that the exposure to x-rays would have an
impact on recombination rates of Sordaria.
Conclusion:
The authors hypothesis that the Sordaria Fimicola that is exposed to
the x-ray treatment will express a higher frequency of recombination
compared to the control Sordaria Fimicola is supported by the data.
The support is derived from the different frequency of recombination
rates in the control 52%, to the treatment 76.6%.

Research Questions:
In the authors experiment, crossing over occurred between the two
different spore colors strains of Sordaria, tan and wild type. This cross
over rate is what, in the experiments, was dubbed the recombination
rate. This recombination rate was simple to observe, since the trait
being observed for recombination was the color of the spores. To
observe the ascus sacs that held the asci posed a challenge in
preparing the samples to be observed under the microscope. To break
apart the perithecia, to release the ascus sac, a squash has to be
performed. There is no exact technique to this processes; it is just trial
and error. The author used a pencil eraser to apply pressure to the
Sordaria, once it was on the microscope slide to break the perithecia
apart. It took a few tries because too much pressure breaks open the
ascus sac and too little causes no break in the perithecia. Once the
challenges of the squashes were taken care of, the author could score
the asci types, to figure out recombination rates. The cross-over
frequency of our control Sordiaria(grown under standard laboratory
growth conditions) was 52%. The Sordiaria grown under the x-ray
radiation (Treatment) displayed a much higher cross over rate of
76.6%.
Sources of error:
A potential source of error could be from when the author counted the
asci types that were revealed after the squash. The author only

counted twenty asci types for the control and twenty for the treatment.
There were thousands of asci on each squash, so twenty is small
number. This potential source of error was slightly corrected by having
twenty-three sets of twenty counted adding up to a total of 460 asci
counted for the control and treatment. Another source of error could
be if all the treatments samples were not exposed to the same level of
x-ray radiation. Uneven exposure to radiation could cause the cross
over rates to be different between the treatment set of Sordaria. It is
important to keep the treatment uniform in an experiment. The author
is not aware of these factors specifics, but this is a potential source of
error.

Significance of the experiment:


This experiment investigated the affects of x-rays on recombination
frequency in a model organism, Sordaria Fimicola. The data from this
experiment suggest that x-rays have a strong affect on cell division by
increasing crossover frequencies (Recombination). The significance of
this data is that x-rays are not good for a body since it has such a
strong affect on recombination rates in cell division. Also, when an
organism is exposed to a stressful environment it is either trying to
adapt through natural selection or the mechanism for mitosis and
meiosis are altered resulting in higher frequencies of recombination in
cell division.

Future Experiments:
A future experiment to further test the same dependent variable could
be to see if other unnatural environments, across the electromagnetic
spectrum, cause the same results, and if so, at a greater or lesser
extent. Environments such as radio, micro, infrared, gamma,
ultraviolent...etc., and other wavelengths could be used. The intensity
and or duration of the x-rays that the Sordaria Fimicola was exposed to
could be changed. A completely new experiment that could arise from
the experiment the author conducted could be derived from the
questions; why did the x-rays have that affect that they did, what
mechanism of cell division did the x-rays affect, what other factors
have this affect on the Sordaria, and are there any instances in nature
that this happens? To examine some of these questions it would take a
much more complicated experiment and equipment. A way to monitor
the Sordaria while it was being exposed to the x-rays and dividing
sexually would be needed.

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Sources:
"Ascus | Fungal Reproduction." Encyclopedia Britannica Online.
Encyclopedia
Britannica. Web. 26 Oct. 2015.
Burpee, D., Cyr, R., Hass, C., Ikis, D., Richter, K., Ward, A. and D.
Woodward, eds. A
Laboratory Manual for Biology 110 Biology:
Basic Concepts and Biodiversity.
2015. Department of Biology, The
Pennsylvania State University, University Park, PA
Lodish, Harvey. DNA Damage and Repair and Their Role in
Carcinogenesis. DNA Damage and Repair and Their Role in
Carcinogenesis. U.S. National Library of Medicine, 2000. Web. 26 Oct.
2015.
Saleem, Muhammad, and Bernard Lamb. Genetics. Inherited
Differences in Crossing Over and Gene Conversion Frequencies
Between Wild Strains of Sordaria
Fimicola From Evolution Canyon
Genetic Soceity of America, 1 Dec. 2001. Web. 24 Oct. 2015.
Volk, Tom. "Sordaria Fimicola, a Fungus Used in Genetics-- Tom Volk's
Fungus of the
Month for March 2007." Sordaria Fimicola, a Fungus
Used in Genetics-- Tom Volk's Fungus of the Month for March 2007.
2007. Web. 26 Oct. 2015.

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