Selective

Advantage of Eye Mutations Drosophila melanogaster

Biology 315
Genetics Lab
Fall 2015
Sun chi
Thursday 09:30-12:20pm


Carletti, Sam
Carney, Jeffory
Miller, Chis

Carletti, Carney, Miller 2

Abstract:
Our hypothesis was that the red-eyed, wild type population of fruit flies
(Drosophilia melanogaster) would have a selective advantage over eye-color
mutants in some part of the reproductive cycle. We compared three populations
(red-eyed [wild type], white-eyed, and sepia-eyed) through a cycle of egg laying
through adulthood, comparing both numbers of eggs laid, and percentage of eggs
which survived, to adulthood. We placed two breeding pairs into each tube (three
tubes per population; for a total of 36 flies) into separate vials containing a small
amount of fly maiden. Once we saw eggs, we removed the parents, and counted how
many eggs each population had laid. Then we monitored the survival rate from egg
to larva to pupa to adult. We used a T-test to compare our mutant phenotype
survival rates to our wild-type survival rate. The T-test failed to support our
hypothesis, because we failed to reject the null hypothesis that there was no
significant difference between the populations survivability rates. There was,
however, a difference in terms of the number of eggs laid, and while we could not
confirm that a selective advantage lay here, the data raised the possibility that it
could. During the course of the experiment, we encountered several problems,
including the desiccation of two of the three tubes for one population.

Carletti, Carney, Miller 3

Introductions:
Drosophila melanogaster eyes are a focus of attention for defining
mechanisms of development. Since visual systems are not strictly necessary for
survival, the flies are able to develop mutations at the alleles that control eye color,
and can even develop reduced eyes or lose their eyes entirely. The eye consists of a
regular lattice structure comprised of 800 repeating 22-cell units. In a large number
of investigations geneticist have found that mutant fruit flies have a lower fitness
when compared to the wild type. Connolly found that when placing any kind of
mutant genes in a population with wild types flies, the mutant genes are either
eliminated or maintained at a very low frequency. (Connolly 1969) Harland found in
his study that most mutants of Drosophila melanogaster are at a disadvantage
compared with the wild type in the environmental conditions provided by nature.
(Harland 1935)
We are looking deeper into the effects of eye mutations on the ability to
survive through the different development stages. This is important because it could
help explain why the red-eye phenotype is the wild type. There are three main
stages for a fruit fly to mature. The first is eggs. Eggs are about 1/2mm in length and
are shaped like grain of rice. Within twenty-four hours eggs hatch and become larva,
which will then go through two molts to begin the pupation stage. Once the larva
goes through the second molt they will become pupa. Later, they will emerge from
the pupa as adults. The entire process from conception to adulthood ranges from
anywhere between nine to twelve days. (Image A) Our hypothesis is that the eye
mutations will decreases fitness in terms of number of eggs laid or survivability

Carletti, Carney, Miller 4

from egg to adult in our two mutation groups when compared to our wild type
population. We developed our hypothesis this way because we assume there is a
selective advantage for the red-eyed phenotype. (i.e. because it is the wild type)
Materials and Methods:
We set up three see-through vials with foam stoppers containing three
isolated populations (A, E, and G) of Drosophila melanogaster which each had
different phenotypes. Population A contained a white-eye mutation, population E
was the red-eyed wild type, and population G was a sepia-eyed mutation. Because
population E is a wild type we used it as our control group to compare with the
mutants. The vials contained the populations of D. melanogaster as well as a food
source in the bottom of the vial to keep the organisms alive, which was made up of
Drosophila Formula 4-24, water, and yeast. We removed the adults from each of the
vials after they laid eggs. Then waited till our eggs hatched to larva and larva, then
pupae molted into adults. We fly napped the populations to allow us to separate
males from females and put a male and female of the same phenotype as a couple
into a vial. Two breeding pairs of the same phenotype were placed in the vials on
October sixth, 2015 at 11:30. The vial contained a thin layer of food at the bottom so
we could more easily observe the eggs and make accurate measurements of larva.
We repeated the previous step with another two vials so we had a total of three vials
with two breeding pairs for each of the three different populations, while keeping all
the containers at the same room temperature at 76 degrees Fahrenheit. We checked
on a regular basis for eggs to form in the food medium. Once the eggs were
observable we counted them and waited till they hatched to larva as well as

Carletti, Carney, Miller 5

removed the parents from the vial so no further procreation would occur. From
there we counted the larva and found the percentage that survived hatching and did
the same that survived molting to an adult. Once an adult is reared, we recorded and
removed the adult from the vial being studied. Our sample size begun with six
breeding pairs for each population, so a total of thirty-six fruit flies yet will increase
due to the unknown reproduction rate and mortality rate of the off spring. Once we
collected our data we compared our results to our wild type populations. From
there we can either see if one or more of our populations phenotype is more
advantageous than another by comparing percentages at each life cycle stage and
using a two tailed T-test to see if our results are significant. This will allow us to
statistically accept or reject our hypothesis that the eye mutations will decreases
fitness in terms of number of eggs laid or survivability from egg to adult in our two
mutation groups when compared to our wild type population.
Results and Discussion:
We examined two variables: the number of eggs laid, and the viability of
those eggs to adulthood. The three populations did lay different numbers of eggs.
The white-eyed populations (A) laid an average of 68 eggs between the three vials.
Populations G (sepia-eyes) laid an average of 59 eggs among the tree vials. The wild
type E population laid an average of 125 between the three vials (about twice as
many).
It has been established by prior research that homogamic mating between
sepia-eye colored flies produces fewer eggs (Stani, 2005), which mirrors the

Carletti, Carney, Miller 6

results we found in our G population. While we have no scholarly data on the whiteeyed population (A), their numbers might follow a similar tendency.
The other variable we looked at was egg viability, measured as survival rate
from egg to adulthood. Here the results were surprising, but also problematic. The
wild type and white -eyed phenotype flies had similar rates of survival. One of The
sepia phenotype, however, seemed to have a high egg to adult survival rate.
However, the sepia survival rate tubes had problems. Only one of the tubes
had the extremely high survival rate (G1). The other two had low rates, but that is
probably because these two tubes desiccated during the course of the experiment.
G2 desiccated during the larval stage on October 13th (leading, presumably, to a low
pupae number). G3 desiccated during the pupal stage on October 16th. We added
water to desiccated tubes but it is hard to know how that affected the experiment.
We used a T-test to look for (if any) significant difference in our populations.
For our T-test we compared our mutant populations to our controlled group E, wild
type. (Table A) We calculated t-values for each crucial life stage; egg to lava, lava to
pupae, and pupae to adult. For population A (white-eye mutation) our t-value was
0.314 for egg to lava, lava to pupae was -0.756, and pupae to adult was 1.57. (Table
B) T-values for population G consisted of 1.52 for egg to lava, 0.733 for lava to
pupae, and 0.210 for pupae to adults. We used a degree of freedom of two and
compared those statistical values to a two-tailed T-table at a p- value of 0.05. The
critical value for a t- value is P> 0.05 is 4.303. We can see that all of our values for
both mutating populations are P> 0.05. We reject our hypostasis that there would be
a lower survival rate for the mutant population, thus accepting the null hypostasis

Carletti, Carney, Miller 7

that there is no significant differences in the survival rate between wild type and eye
color mutant populations.
Overall we rejected our hypothesis statistically. Still, based on problems that
arose during the experiment, and the small population sizes studied, there is still
room for questions, and several interesting possibilities did come up.
The first possibility comes from the low egg numbers, but high survivability
numbers for one of the G population. An interesting question to study further would
be whether laying fewer numbers of eggs means that those eggs represent a higher
energy investment on the part of the fly, and are therefore more viable. If that higher
egg viability leads to higher numbers of sepia colored flies making it to adulthood
(which happened in our experiment), this would contradict our hypothesis. The
sepia mutation would then have a selective advantage over the wild type. Obviously,
this possibility is very understudied, but the data does raise the question.
It is unlikely that this possible selective advantage would be directly related
to eye-coloration, because eye color probably has little to do with the number of
eggs or their viability. It is more likely that the genes, which affect eye color, would
interact through epistasis with the genes, which control egg production and early
stage development. However, a different type of study would be required to learn
what genetic mechanism underlies these results.

In short, the experimental data is conclusive that there is not statistical

significance of survival rates between wild type and eye color mutants. However,
further research would be required to confirm whether or not white and sepia flies
do indeed lay fewer eggs, and if the sepia eggs are more viable. If those results were

Carletti, Carney, Miller 8

confirmed, further study could go into what genetic mechanism underlies the
observed results.

Carletti, Carney, Miller 9

Experimental Results

Egg Eggs Percent TLarva Percent TPupae Percent TLarva
Pupae
Total Died (%)
Value
Died (%)
Value
Died (%)
Value
72
35
48.6
37
11
29.7
26
8
30.8


180
60
33.3 0.0000 120
46
38.3 0.000
74
35
47.3 0.0000
124
28
22.6
96
32
33.3
64
29
45.3

Table A: Our Experimental data of our controlled group, population E (wild type)


Egg Eggs Percent TTotal Died (%)
Value
67
54
82

16
25
20

23.9

46.3 0.3142
24.4

Larva
51
29
62

Larva Percent TPupae Percent TPupae

Died (%)
Value
Died (%)
Value
23
6
8

45.1

20.7 -0.76
12.9

28
23
54

11
15
41

39.3
65.2 1.5768
75.9

Table B: Our Experimental data of the white-eyed populations and T-test values
against the wild type population


Egg Eggs Percent TTotal Died (%)
Value
64
51
63

8
16
11

12.5

31.4 1.5266
17.5

Larva
56
35
52

Larva Percent TPupae Percent TPupae

Died (%)
Value
Died (%)
Value
7
30
26

12.5

85.7 0.733
50.0

49
5
26

9
3
14

Table C: Our Experimental data of the sepia-eyed populations and T-test values
against the wild type population

18.4
60.0 0.2106
53.8

Carletti, Carney, Miller 10

Image A: Summarizing the life cycle of a fruit fly by depicting their different physical
stages before adulthood.

Carletti, Carney, Miller 11

Bibliography:
Connolly, K., Burnet, B., & Sewell, D. (1969). Selective Mating and Eye
Pigmentation: An Analysis of the Visual Component in the Courtship Behavior of
Drosophila melanogaster. Evolution, 23(4), 1-12. doi:10.2307/2406852

Harland, S. (1935). Advantage of White Eye Mutant of Drosophila
Melanogaster Over the Wild Type in an Artificial Environment. Retrieved October
20, 2015, from http://www.nature.com/hdy/journal/v12/n1/pdf/hdy19582a.pdf

Leiserson, W., & Benzer, S. (1998). Dual functions of the Drosophila eyes
absent gene in the eye and embryo. Retrieved October 20, 2015, from
http://www.sciencedirect.com/science/article/pii/S0925477398000525

Stani, S. (2005, September 1). Mating success of wild type and sepia mutants
Drosophila melanogaster in different choice. Retrieved October 20, 2015, from
http://www.ncbi.nlm.nih.gov/pubmed/16440285