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c
O
Katy Budden, Chris Karnes, Justin Kazee, Brittany Milhoan
c
O
is a gold standard model organism for genetic studies. The
purpose of this experiment was to verify the status of the white eye mutation as a sex-linked
condition. In this experiment, wild-type (red-eyed) females were crossed with white-eyed males.
The resulting offspring (F1 generation) were crossbred to produce the F2 generation. This F2
generation was then crossbred to produce the F3 generation. It was hypothesized that the results
of these crosses would fit a 3:1 phenotypic ratio in the F2 generation and a 13:3 phenotypic ratio
in the F3 generation. Chi square analysis of the data yielded a p value << 0.001, thus rejecting
the hypothesis. The actual results showed that all individuals in the F1, F2, and F3 generations
displayed the white-eyed phenotype. This likely occurred as the result of the mating between a
single white eyed female with the white eyed males. There is also the possibility of the wild-type
females being heterozygous for the white-eyed mutation.
The white eye phenotype in O
is due to a recessive sex-linked
mutation located on the X chromosome. The genotype of a female fly expressing this trait is
white-eyed male with a virgin wild-type female resulting in an F1 generation made up of 1,237
wild-type offspring and three white-eyed flies, all of which were male. The F2 generation
consisted of 2,459 wild-type females, 1,011 wild-type males, and 782 white-eyed males
(Morgan). In this experiment, wild-type female flies were crossed with white-eyed male flies in
order to observe the phenotypic trends of the white-eyed mutation throughout the subsequent
generations. Chi square analysis was used to analyze the data from the F2 and F3 generations. It
was hypothesized that when wild-type females are crossed with white-eyed males, the resulting
offspring ratio of the F2 generation would have a 3:1 phenotypic ratio of wild-type flies to white-
eyed flies.
Six virgin wild-type (red-eyed) females were crossed with six white-eyed males to
produce the F1 generation. The flies were incubated for a week in a vial with growth medium
and allowed to mate. The following week, the P1 generation was removed from the vial to
prevent breeding between generations, and the larvae were incubated for another week to enable
further development. Once the F1 generation had developed into adults, they were anesthetized
and counted according to sex and phenotype. The F1 generation was then introduced into a new
vial and allowed to breed to produce the F2 generation. These steps were repeated to produce
and count the F2 and F3 generations. The results were analyzed using Chi square analysis to
obtain a P-value.
Table 1 ± Expected P1 Cross
Male (P1)
Female (P1) Xw Y
X XXw XY
X XXw XY
The white-eye mutation was not expected to be expressed in any of the F1 offspring.
Table 2 ± Expected F1 Cross
Male (F1)
Female (F1) X Y
Xw XXw XwY
X XX XY
The white-eye mutation was expected to be expressed in one-fourth of the offspring (one-half of
Expected Observed
Red-eyed females 3.5 0
Red-eyed males 3.5 0
White-eyed females 0 0
White-eyed males 0 7
Table 5 ± Actual Results for F2 Generation
Expected Observed
Red-eyed females 24.5 0
Red-eyed males 12.25 0
White-eyed females 0 33
White-eyed males 12.25 16
Table 6 ± Actual Results for F3 Generation
Expected Observed
Red-eyed females 56.875 0
Red-eyed males 48.75 0
White-eyed females 16.25 55
White-eyed males 8.125 75
Expected Observed (O) Expected (E) Deviation (D) Deviation2 (D2) D2/E
Ratio
¾ 0 wild 105.6 wild -105.6 11,151.36 105.6
¼ 130 white 24.4 white 150.6 11,151.36 457.0
Total: 562.6
p value <<0.001
In both of the above tables, the p value was highly insignificant thus rejecting the hypothesis.
O
In the first two weeks of the experiment, the wild-type females and white-eyed males
were crossed. By the third week, the P1 generation had produced an F1 generation containing
seven white-eyed flies. It was believed that there were both males and
flies males and six-white eyed females were taken from another group
that had done the same cross. The following week, it was discovered
that all of the flies were deceased. This was most likely due to excessive
use of FlyNap®. Therefore, an additional six white-eyed males and six white-eyed females were
obtained from the same group. Because there were no wild-type alleles present in the population,
it was expected that there would not be any offspring exhibiting the wild-type phenotype.
The results from each filial generation showed 100 percent penetrance of the white-eyed
showed that the obtained results (P<<0.001) were not due to chance, thus rejecting the proposed
After aberrant results were obtained for the filial generations (all white-eyed individuals),
it was originally theorized that heterozygous females for the white-eyed mutation were present in
the wild-type vial. Once these flies were crossed with the white-eyed males, both white-eyed and
wild-type flies would have been produced, each yielding 50% (Farrell). However, the
probability that only white-eyed flies were produced would be highly unlikely. Another proposed
theory was that there was at least one white-eyed female in the group of white-eyed males. If this
cross occurred, 100% of the offspring would have portrayed the white-eyed mutation, although it
is unlikely that only the white-eyed female would have produced the entire F1 generation.
However, if both of these possible theories occurred simultaneously, the likelihood of having
strictly white-eyed offspring would be significantly higher. The lack of wild-type offspring in
Based strictly on the results of this experiment, the genetic inheritance of this trait can be
Hunt Morgan and many others, it is a known fact that the white-eyed mutation is a sex-linked
recessive. So why did the results suggest a completely different inheritance pattern? Though it is
unclear what the exact cause of this differentiation is, there are several possible causes, including
but not limited to the genotypes of the supplied flies and the possibility of the addition of an
Farrell, Courtney. "Thomas Hunt Morgan: Chromosomes and Heredity." (2006) 1 Dec
2008 <http://web.ebscohost.com/ehost/detail?vid=4&hid=101&sid=41506e4a-
8da6-4a1f-b853-
0ffc8cdcef65%40sessionmgr102&bdata=JnNpdGU9ZWhvc3QtbGl2ZQ%3d%3d
#db=sch&AN=20277707>.