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c 

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

XwXw and the male genotype is XwY (³Simple

Mendelian«´). This mutation was first discovered by T.

H. Morgan in 1910 through his demonstration of the

correlation between the inheritance of this mutation and

the transmission of the X chromosome (Green). This was

proven again in 1916 with C.B Bridge¶s publication in volume I of 



, discussing the

nondisjunction of the O


  
X chromosome (Green). Morgan crossed a spontaneous

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

the male offspring and none of the females).

Tables 3.1-3.4 ± Expected F2 Crosses



Table 3.1 Table 3.2

Male (F2) Male (F2)
Female (F2) Xw Y Female (F2) X Y
X XXw XY X XX XY
Xw XwXw XwY Xw XXw XwY


 Table 3.3 Table 3.4

Male (F2) Male (F2)
Female (F2) Xw Y Female (F2) X Y
w
X XX XY X XX XY
w
X XX XY X XX XY

The white-eye mutation was expected to be expressed in three out of sixteen offspring.
The above tables show all possible crosses resulting from the F2 generation.

Table 4 ± Actual Results for F1 Generation

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


Table 7 ± Chi Square Analysis of F2 Generation



Expected Observed (O) Expected (E) Deviation (D) Deviation2 (D2) D2/E
Ratio
¾ 0 wild 36.75 wild -36.75 1,351 36.8
¼ 49 white 12.25 white 36.75 1,351 110.3
Total: 147.1
p value <<0.001


Table 8 ± Chi Square Analysis of F3 Generation

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

females in the vial. After another week, further investigation showed

that there were actually seven males and no females. Therefore, an F2

generation was not produced. To further the experiment, six white-eyed

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

phenotype (Tables 4-6). The F1 generation was

expected to exhibit strictly the wild-type

phenotype (heterozygous females, wild-type

males), while the F2 generation should have

exhibited a phenotypic ratio of 3:1 wild-type to

white-eyed individuals, and the F3 generation was

expected to fit a phenotypic ratio of 13:3 wild-

type to white-eyed individuals (Tables 1-3.4). In „ 



„ 
 

contrast, the actual results for all generations


showed exclusively white-eyed flies. Chi square analysis of the data from the F2 generation

showed that the obtained results (P<<0.001) were not due to chance, thus rejecting the proposed

hypothesis (Tables 7-8).

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

this scenario could also be attributed to the rather small population.

  

Based strictly on the results of this experiment, the genetic inheritance of this trait can be

defined as an autosomal dominant mutation. However, based on the experiments of Thomas

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

incorrectly sexed fly.



    

Morgan, TH: (1910) ³Sex Limited Inheritance in O .´ 



, 3298120:120-122

"Simple Mendelian Genetics in O ." http://math.hws.edu. 17 Nov 2008


<http://math.hws.edu/javamath/ryan/Genetics1.html>.

Green, M.M.. "The "Genesis of the White-Eyed Mutant" in O 


  
: A
Reappraisal." Genetics 142Feb 1996 329-331. 1 Dec 2008
<http://www.genetics.org/cgi/reprint/142/2/329>.

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>.