The Relationship between Meiosis and Genetic Diversity in Sordaria fimicola

Name: Emma Schwendeman

Course: Biology 110H, Section 001H
TA: Brandon Follick

Introduction
In Sordaria fimicola, meiosis is the essential form of reproduction for these organisms.
Meiosis is unique due to the fact that it allows the process of crossing over between non-sister
chromatids of two chromosomes to occur, which in turn allows Sordaria firmicola to create
genetic diversity in its species. Evidence of this phenomenon is illustrated through the work on
Evolution Canyon in Israel. The area contains two slopes, noted as the South-Facing Slope and
North-Facing Slope, beholding opposite environments to one another. On the SFS, the
environment consists of more extreme factors such as high radiation to the area, higher
temperatures, and much more desert-like conditions. On the NFS, there tends to be a more
temperate and cool environment. These differences bring to light many questions about how this
came about. Researchers have now wondered after observing extreme differences in conditions
on each slope if there are differences in the cross-over frequencies of these organisms (Hass et.
al, 35-37).
After performing crosses between mutants and wild type, scientists hypothesized that
organisms, in particular Sordaria, living on the SFS contained a higher cross-over frequency
compared to the NFS. This lab works to test this hypothesis, also working to provide the baseline
material to standardized cross-over frequencies under optimal lab conditions. This information
can be used as a control or resource in understanding if environmental stresses play a bigger role
(Hass et. al, 35-37).
As stated before, meiosis is a crucial part to spore color and its arrangements. Genetic
diversity is achieved through meiosis when the chromatids of one chromosome are crossed over
with chromatids of the other chromosome (Cyr). Diversity can be seen in the three patterns
obtained from the Sordaria. The first is a 4:4 ratio of spore color in which crossing over does not

occur between the mutant, tan coloring, and wild type, black coloring, yet independent
assortment can give two order of the colors. The second is a 2:4:2 ratio where crossing over
occurs in prophase I of meiosis between the two non-sister chromatids, the inner of either the
mutant or wild type and the outer chromatid of the other. Independent assortment plays a role in
this pattern. The last type of cross-over is the 2:2:2:2. Occurring in prophase I, crossing over
occurs between non-sister chromatids, the inner non-sister chromatids, with independent
assortment yet again determining which color goes first. These spore patterns occur due to the
Sordaria life cycle in which they make perithecium through mating the two hyphae together in
sexual reproduction. From there, they form an ascus after going through meiosis and potentially
crossing over (Hass et. al, 48-49, 52).
Finding these crosses and information is important in order to answer the questions that
have been given. These objectives range from crossing over between the spore color strain and
the mutant to solving the map distance predicted by the cross-over frequency. Even more, this
lab requires the class to explain challenges that arose, either in preparing through standard
procedure of mating different strains or in squashing the perithecia in order to get asci to observe.
Taking it one step further, it works to explain how one might solve the challenges within the lab.
Lastly, this lab requires the class to determine the cross-over frequency between the spore color
gene and the centromere (Hass et. al, 37).
A mating agar plate was prepared by dividing the plate into four separate quadrants. The
two different strains, wild type and mutant, are placed diagonally from each other and then
incubated for two weeks in order to allow mating to occur. This is crucial in order to obtain
perithecia and asci with multitudes of crosses for observations. After mating has occurred and the
perithecia has been squashed to observe spore color patterns, recording the amount of each type

will help in calculating the cross-over frequency, either an individual types cross-over frequency
or overall cross-over frequency. Map distance can be calculated using the overall cross over
frequency, dividing it by two since two non-sister chromatids are only involved in it. The
crossing over events only occur between two non-sister chromatids to create these patterns. This
explains the reasoning that map distance has to be divided by two, whether these chromatids are
the inner non-sister chromatids or the inner and outer non-sister chromatids (Hass et. al, 42, 5559).
Materials and Methods
To set up crosses, the two strains, wild type and mutant, were provided. The next step
was to divide the plate with the marker into 4 sections and labeled tan and wild type, making
sure that the same strain was diagonal from one another. To ensure a valid mating plate to record
the best results, it is necessary to wipe off the work surface with disinfectant wipes as well as
disinfect the scalpel. Carefully, placing two squares of agar containing the fungal hyphae from
either the mutant or wild type was to be put in their labeled squares. The hyphae were placed
face down on the agar. After disinfecting the scalpel for a second time, the process was repeated,
using the other strain. These cultures were incubated at room temperature for approximately two
weeks in order to allow proper mating to occur (Hass et. al, 42).
Once mating occurred, an inoculating loop was utilized. Perithecia was scraped from the
center of a dividing line on the plate where the two strains met, placing them then on a
microscope slide that contained the mixture of a few drops of water and a bit of the perithecia.
This combination was covered with a coverslip. Using a small bit of pressure from a pencil
eraser, the coverslip was pressed down to release asci from perithecia in the process called

squashing. Squashes were then observed under a microscope in order to analyze the asci patterns
(Hass et. al, 55-58).
After observing the asci patterns, either of 4:4, 2:2:2:2, or 2:4:2, the information was
recorded in the provided tables in the lab manual. Calculating the recombinant frequency was
completed, utilizing the equation of the recombinants over the total number of patterns, or
(B+C)/(A+B+C). Map distances were calculated as half the number of percent crossover
frequency. Dividing by two is an important step because when two chromosomes go through
crossing over, only the two non-sister chromatids are being crossed over and not the whole
chromosomes (Hass et. al, 55-59).
Results
Table 1. Number for Each Type of Spore Pattern for Each Set of Data
For this table, counts of each spore pattern were taken for ones individual sample, the table
groups samples, the sections samples, and the entire courses sample respectively.
Number

Number

Number

of Type A

of Type B

of Type C

(4:4) asci

(2:4:2)

(2:2:2:2)

Individua

asci
8

asci
5

l Data
Small

15

15

10

180

165

179

Group
Data
Table
Combine

d Section
Data
Combine

10,512

7811

7855

d Course
Data

Table 2. Total Numbers for Asci Patterns and Numbers for Recombinant Asci Patterns
To further help calculations, the sum of all three patterns and sum of the recombinants were
calculated for each grouping.

Individual Data
Small Group Data Table
Combined Section Data
Combined Course Data

Total Number of Asci

Total Number of

(A+B+C)
20
40
523
26, 178

Recombinant Asci (B+C)

13
25
344
15,666

Table 3. Cross-Over Frequency Data for Each Recombinant Pattern

With two asci patterns developed from crossover, each patterns individual frequency was
calculated for each grouping.

Individual Data
Small Group Data Table
Combined Section Data
Combined Course Data

Crossover frequency of

Crossover frequency of

Type B Asci (2:4:2)

Type C Asci

(B/total)
0.4 40%
0.375 37.5%
0.31549 31.549%
0.29838 29.838%

(2:2:2:2)(C/total)
0.25 25%
0.25 25%
0.34225 34.225%
0.30006 30.006%

Table 4. Overall Cross-Over Frequencies for Each Data Group

The overall crossover frequency was calculated and recorded for each grouping.

Individual Data
Small Group Data Table
Combined Section Data
Combined Course Data

Crossover frequency overall ((B+C)/total)

0.65 65%
0.625 62.5%
0.65774 65.774%
0.59844 59.844%

Sample calculation of map distance on genome:

Crossing over occurred between the spore color gene and the centromere, as seen through
the combinations of 2:4:2 and 2:2:2:2 recorded. The usage of this data can then be utilized to find
the map distance on a genome of the cross over, demonstrating that crossing over occurred
between the gene and the centromere. The average cross-over frequency was calculated to be
59.844% for the combined course data. Using the formula for cross-over frequency over 2, the
map distance that was calculated is 29.922 cM, used from the combined course data. Here is a
sample calculation to demonstrate this calculation:
Combined Course Data
Frequency: 15, 666/26, 178= 0.59844
To find map distance= C.O.F./2= 0.59844/2x 100%= 29.922cM (Hass et. al., 55-59)
Discussion
After calculating and observing the data and trends within them, there are a few new
statements pertaining to what was learned from this lab. To start out, crossing over does occur
between the spore color gene and the centromere. Evidence can be found in the lab because
when gathering results from the mating plate, there are a number of recombinant types of asci
that arose. Furthermore, when under optimal laboratory growth temperatures, the crossing over

frequency is generally consistent. This can be shown in the tables and calculations of cross-over
frequency which shows that with each sample getting bigger, the frequency stays relatively the
same (Hass et. al, 60).
In the experiment, the individual plate contained some rather unexpected findings within
it. The first was that a white film covered the agar plate, which was later learned to be a fungal
infection. Although it had been believed that the proper steps of sterilization were taken, the
finding suggested that maybe not all necessary steps were properly carried out. Techniques such
as washing hands or ensuring critical sterilization to every item could have prevent this from
occuring. In addition to that, the asci sometimes contained patterns that were not common
patterns discussed in class. At points, there would be asci containing spores that were 2 black and
6 tan. Most likely, mutations for the gene could have created an abnormal pattern. As well, the
functions of crossing over and independent assortment could have been a contribution to this
abnormality (Hass et. al, 60).
The material attained from the lab can help in future experiments in researching what the
cross over frequency is for certain forms of environmental stresses. The lab helped in presenting
a baseline of information for cross-over frequency for optimal lab conditions. Knowing that the
SFS side at Evolution Canyon had a higher frequency, a new hypothesis can be formed and
tested to analyze if environmental stresses affected the cross-over frequency. This information
can then show that meiosis plays a huge role in genetic diversity and helps in allowing organisms
to adapt to the environments they live in. Without the baseline information, the research currently
conducted could not proceed further as efficiently.
References

Cyr, R. 2002. Heredity and Life Cycles. In, Biology 110: Basic concepts and biodiverity course
website. Department of Biology, The Pennsylvania State University.
http://www.bio.psu.edu/
Meiosis and Genetic Diversity in the Model Organism, Sordaria. Written by Hass, C., Richter,
K., and Ward, A. 2014. Department of Biology, The Pennsylvania State University,
University Park, PA.