Duncan Ackerman

Biol 110, Sec. 911

11/6/2013
Lab Report #1


Meiosis and Genetic Diversity in Sordaria fimicola

Introduction (Abstract):

Evolution canyon has a North and a South-facing slope with drastically different
environmental conditions. The North facing slope has a European type climate while
the South facing slope has an African type climate. The differences in these climates
likely produce variances in crossover frequencies between the Sordaria that live on
each slope (Nevo 2009). Before the discoveries at Evolution Canyon, scientists did
not believe that environment had an impact on organisms at a molecular level via
processes like cross over frequency during meiosis. A harsher climate could yield
greater crossover frequencies due to the need for environmental adaptations for the
survival of the Sordaria (Lamb, et al. 1998; Salemm, et al. 2001). A team from
Imperial College in London found that the crossover frequencies were greater in
Sordaria grown on different sides of the slope than those grown on the same side,
which gives evidence to support the aforementioned possibility.

In order to determine whether environmental conditions have an effect on
crossover frequencies, baseline information is needed. To figure out the answer to
this question, Sordaria must be grown in optimal laboratory condition and
crossover frequency must be recorded to compare to the frequencies in those living
on Evolution Canyon. Three different types of asci occur in Sordaria. The spores can
be arranged in tan to wild type ratios as follows: 4:4; 2:2:2:2; and 2:4:2. This
depends on the frequency of crossing over in the samples. If a 4:4 ratio is produced,
no crossing over has occurred. In 2:2:2:2 and 2:4:2, crossing over has occurred. In
2:2:2:2 only basic crossing over occurs while in 2:4:2 the law of segregation and
independent assortment allow for chromosomal position switches during meiosis II.

Overarching Research Question: How does environmental stress effect cross-over
frequency, structures associated with meiosis and gene expression related to
meiosis over time?

Brief Overview of Procedures: Wild type Sordaria and tan Sordaria were grown in
four quarters of a petri dish and were allowed to cross in bordering areas and then a
squash was performed and cross-over frequencies were measured. The possible
frequencies are as follows: 2:2:2:2, 2:4:2, and no crossing over at all, 4:4.

Methods

Crosses were set up by planting Sordaria in an auger with 4 sections: 2 with tan
diagonal from each other and 2 with wild-type also diagonal in a circular plate. They
were allowed to grow for 2 weeks and breed with each other at the borders of each
types planting area. Squashes were then prepared by first extracting asci from the
areas where each type of Sordaria met on the plate. Then a drop of water was added
to a slide and the asci were placed on it. Next the slide was examined under the
microscope and the top of the slide was tapped to burst the asci. Last, the spore
patterns of the burst asci were examined to determine where recombination
occurred. The cross-over frequencies were calculated by comparing the number of
recombinant spore groups to the number of non-recombinant spore groups. The
number of recombinant spores(331) was divided by the total number of spore
groups examined(500) and it was determined that there was a 66.2% recombinance
rate. 66.2 was then divided by two to get a map distance of 33.1.



Results


Data Tables Attached


Patterns Observed: Crossing over does occur between spore color gene and the
centromere. We observed a crossover frequency of 66.2%. The over crossover
frequency between spore color gene and the centromere in Sordaria grown under
laboratory conditions was 66.2%. The calculated map distance between the spore
color gene and the centromere is 33.1.



Discussion

We learned that crossing over does indeed occur in sordaria with a rate of 66.2%
and that the map distance of the color gene to the centromere is 33.1.

We were able to answer the following questions through our research:
1. What challenges arise in using the provided procedure for mating different
strains of Sordaria?
The Sordaria strains may not mate correctly in the dishes. Sometimes, if the
tan and wild type do not mate with each other at the borders of where each
type is placed in the dish, then crossing over will not be able to be sufficiently
observed because it will not occur.
2. What challenges arise in preparing squashes of perithecia for scoring asci using
the provided procedure?
The squashes were difficult to create at first. The pressure applied to the
slide must not be too great or too little and must be applied to a precise
portion of the slide in order to produce the desire results.
3. What evidence demonstrates that corossing-over occurred between the
spore color gene and the centromere?
The fact that the arrangement of the spores in the 2:2:2:2 pattern and the
2:4:2 pattern were observed in significant numbers proves that crossing over
had to occur. If not, all the spores would be arranged in a 4:4 pattern if
mating occurred.
4. What is the crossover frequency of the spore color gene with the centromere
in organisms grown under optimal conditions?
66.2% for our experiment
5. What is the map distance between the spore color gene and the
centromere predicted by the crossover frequency under optimal growth
conditions?
33.1 map units.




The high crossover frequency under optimal laboratory conditions was unexpected.
It would have made more sense if conditions were not optimal since the Sordaria
would need to adapt to an unfriendly environment but this rate of crossing over
should not be necessary when conditions are so conducive to Sordaria survival.
This information could assist further research of cross over frequencies in Sordaria
and could be used as a baseline for the research of these fungi on Evolution Canyon




References:

Lamb, B.C., Saleem, M. Scott, W., Thapa, N. and Nevo, E. (1998). Inherited and
environmentally induced differences in mutation grequencies between wild strains
of Sordaria fimicola from evolution Canyon Genetics 149, 87-99.

Nevo, E. (2009). Evolution in action across life at Evolution Canyon, Israel. Trends
in Evolutionary Biology 1:e3.

Salem, M., Lamb, B.C., and Nevo, E. (2001). Inherited differences in crossing over and
gene conversion frequencies between wild strains of Sordaria fimicola from
Evolution Canyon Genetics 159, 1573-1593.