UW-La Crosse

Investigating Mendelian Inheritance Patterns Relating to the Expression of

Mutations Following a Cross of True-Breeding Drosophila melanogaster

Jessica Nelson

BIO 306 - Genetics

Dr. Amy Yu

November 16, 2017

 Investigating Mendelian Inheritance Patterns Relating to the Expression of Mutations

Following a Cross of True-Breeding Drosophila melanogaster

Abstract

The goal of this experiment was to test Mendel’s laws of inheritance by carrying out a

dihybrid cross of Drosophila melanogaster between true-breeding virgin yellow sex-linked

mutant females and true-breeding cinnabar autosomal mutant males. The cinnabar eye mutation

is in genes located on chromosome two whereas the yellow body mutation is in genes located on

the X-chromosome. This current investigation was focused on introducing the two allele

mutations, cinnabar eyes and yellow bodies, performing a dihybrid cross spanning three

generations, analyzing the resulting phenotypes, and utilizing Chi-square statistics to evaluate the

F2 progenies’ phenotypic ratios. The hypothesis was the dihybrid fly cross of Drosophila

melanogaster between true-breeding virgin yellow sex-linked mutant females and true-breeding

cinnabar autosomal mutant males would be recessive to their wild-types, assort independently,

and follow Mendelian inheritance patterns resulting in male and female phenotypic ratios of

3:1:1:3 of wild-type body and wild-type eyes, wild-type body and cinnabar eyes, yellow body

and cinnabar eyes, and yellow body and wild-type eyes respectively. Following the completion

of the experiment, data of observed and expected phenotypes was used to calculate a Chi-square

value. The resulting Chi-square values were 1.286, 1.031, and 0.5434, which are all less than the

critical value of 7.815, therefore the hypothesis of a F2 phenotypic ratio of 3:1:1:3 failed to be

rejected.
Results

The purpose of this experiment was to test Mendel’s laws of inheritance by carrying out a

dihybrid cross of Drosophila melanogaster between true-breeding virgin yellow sex-linked

mutant females and true-breeding cinnabar autosomal mutant males. A yellow mutation caused

the flies to exhibit yellow bodies while the cinnabar mutation was characterized by bright red

pigmentation in the eyes. The hypothesis was this dihybrid Drosophila melanogaster cross

would follow Mendel’s inheritance patterns, following a 1:1 ratio of females to males in the F1

generation whereas the F2 generation of both male and female phenotypes would follow a

3:1:1:3 ratio of wild-type body and wild-type eyes, wild-type body and cinnabar eyes, yellow

body and cinnabar eyes, and yellow body and wild-type eyes respectively. After the females with

yellow bodies and wild-type eyes were crossed with males with wild-type bodies and cinnabar

eyes, it was observed the F1 generation consisted of mostly males with yellow bodies and wild-

type eyes and females with wild-type bodies and wild-type eyes.

The data collected from the F2 generation resulted in a total of 446 Drosophila

melanogaster with 167 flies wild-type for both traits, 51 flies with wild-type bodies and cinnabar

eyes, 62 flies with yellow bodies and cinnabar eyes, and 166 flies with yellow bodies and wild-

type eyes (Table 1). The observed phenotypic ratio for males and females combined was

calculated to be 3.27 wild-type for both traits, 1.00 wild-type bodied with cinnabar eyes, 1.22

yellow bodied with cinnabar eyes, and 3.25 yellow bodied with wild-type eyes (Table 1). A total

of 209 male flies were further categorized by 75 flies wild-type for both traits, 25 flies with wild-

type bodies and cinnabar eyes, 29 flies with yellow bodies and cinnabar eyes, and 80 flies with

yellow bodies and wild-type eyes (Table 1). Correlating phenotypic ratios for males were

calculated to be 3.00 wild-type for both traits, 1.00 wild-type bodied with cinnabar eyes, 1.16
yellow bodied with cinnabar eyes, and 3.20 yellow bodied with wild-type eyes (Table 1). As for

the females, 237 total flies were counted with 92 flies wild-type for both traits, 26 flies with

wild-type bodies and cinnabar eyes, 33 flies with yellow bodies and cinnabar eyes, and 86 flies

with yellow bodies and wild-type eyes (Table 1). The female observed ratios resulted in 3.54

wild-type for both traits, 1.00 wild-type bodied with cinnabar eyes, 1.27 yellow bodied with

cinnabar eyes, and 3.31 yellow bodied with wild-type eyes (Table 1).

TABLE 1. Observed Drosophila melanogaster phenotypes in the F2 generation

Phenotype Observed Female Observed Male Total Total
females observed males observed observed observed
ratio ratio ratio
Wild type bodies, 92 3.54 75 3.00 167 3.27
wild type eyes
Wild type bodies, 26 1.00 25 1.00 51 1.00
cinnabar eyes
Yellow bodies, 33 1.27 29 1.16 62 1.22
cinnabar eyes
Yellow bodies, 86 3.31 80 3.20 166 3.25
wild type eyes
Totals 237 209 446

The Chi-square test was computed by comparing the previously recorded data of

observed phenotypic ratios from the F2 generation with the expected phenotypic ratios derived

from Mendelian inheritance patterns. The purpose of a Chi-square test is to arithmetically

compute how far results differ from the theoretical expectation and whether the deviations can be

blamed on chance alone (Colavito et al, 2017). In this case, the Chi-square test is determining if

there is a significant difference between the observed and expected phenotypic ratios for the F2

generation of Drosophila melanogaster. The resulting Chi-square value for both female and

males was 1.286 (Table 2). The Chi-square value for only females was 1.031 (Table 3), whereas

the Chi-square value for only males was 0.5434 (Table 4).
TABLE 2. Chi-Square Calculations for Male and Female Drosophila melanogaster phenotypes

in the F2 generation

Phenotype Observed Expected Deviations (o-e)2 (o-e)2/e

(o) (e) (o-e)
Wild type bodies, wild type eyes 167 167.3 -0.3 0.09 0.0005380
Wild type bodies, cinnabar eyes 51 55.75 -4.75 22.56 0.4047
Yellow bodies, cinnabar eyes 62 55.75 -6.75 45.56 0.8173
Yellow bodies, wild type eyes 166 167.3 -3.25 10.56 0.06314
Totals 446 446 1.286

TABLE 3. Chi-Square Calculations for Female Drosophila melanogaster phenotypes in the F2

generation
Phenotype Observed Expected Deviations (o-e)2 (o-e)2/e
(o) (e) (o-e)
Wild type bodies, wild type eyes 92 88.88 3.12 9.734 0.1095
Wild type bodies, cinnabar eyes 26 29.63 -3.63 13.18 0.4448
Yellow bodies, cinnabar eyes 33 29.63 3.37 11.36 0.3834
Yellow bodies, wild type eyes 86 88.88 -2.88 8.294 0.09331

Totals 237 237 1.031

TABLE 4. Chi-Square Calculations for Male Drosophila melanogaster phenotypes in the F2

generation
Phenotype Observed Expected Deviations (o-e)2 (o-e)2/e
(o) (e) (o-e)
Wild type bodies, wild type eyes 75 78.38 -3.38 11.42 0.1458
Wild type bodies, cinnabar eyes 25 26.13 -1.13 1.277 0.04887
Yellow bodies, cinnabar eyes 29 26.13 2.87 8.237 0.3152
Yellow bodies, wild type eyes 80 78.38 1.62 2.624 0.03348
Totals 209 209 0.5434

FIGURE 1. Microscopic images at 4x exemplifying the various phenotypes expressed in

Drosophila melanogaster
Discussion

The purpose of this experiment was to test the accuracy of Mendel’s laws of inheritance

by carrying out a dihybrid cross of Drosophila melanogaster between true-breeding virgin

yellow sex-linked mutant females and true-breeding cinnabar autosomal mutant males. The

parent generation consisted of virgin female with a yellow body mutation crossing with a

cinnabar eye mutation male to result in progeny for the F1 generation. The observation that the

F1 generation consisted of mostly males with yellow bodies and wild-type eyes and females with

wild-type bodies and wild-type eyes supports the claim that the mutations are recessive to their

wild types. These F1 flies cross-bred so that their offspring made up the F2 generation that were

then counted and scored for their phenotypes. As aforementioned, the expected phenotypic ratio

was 3:1:1:3 for wild-type body and wild-type eyes, wild-type body and cinnabar eyes, yellow

body and cinnabar eyes, and yellow body and wild-type eyes respectively. The experiment

yielded a phenotypic ratio for female flies of 3.54:1.00:1.27:3.31 of the same respective

categories while the male flies resulted in a phenotypic ratio of 3.00:1.00:1.16:3.20 of the same

respective categories as well. The observed phenotypic ratio for males and females combined

proved to have similar values of 3.27:1.00:1.22:3.25.

The Chi-square values were then calculated using three degrees of freedom and a critical

alpha value, or p-value, of 0.05, and a corresponding critical Chi-square value of 7.815. It was

found the Chi-square value for both male and female flies combined was 1.286, for females

alone was 1.031, and for just males was 0.5434. The overall calculated Chi-square value of 1.286

and the female flies’ Chi-square value of 1.031 correspond to a p-value between 0.80 and 0.50

while the male flies’ Chi-square value of 0.5434 correspond to a p-value between 0.95 and 0.80.

All three values are lesser than the critical value of 7.815, therefore there was no significant
difference between the expected and observed phenotypes and a hypothesis of a 3:1:1:3 F2

phenotypic ratio cannot be rejected.

Although the hypothesis was not rejected, there are many factors and ways this

experiment could be expanded upon. Other variables that could be investigated include light or

temperature controls, introducing substances, or combining mutations. A potential extension to

this experiment could be administering various foods for Drosophila melanogaster. Specific

amounts of proteins and carbohydrates are required for growth, reproduction, and lifespan.

Previous studies have found trade-offs occur when flies are exposed to conditions with low

levels of carbohydrates and proteins so it would be interesting to see if it could affect phenotypic

expressions of mutations (Trajkovic et al, 2017).

Due to the yellow mutation being sex-linked, a reciprocal cross would yield much

different results. If a male with a yellow mutant phenotype was crossed with a virgin female with

a cinnabar mutant phenotype, the F1 generation would display wild-type for both traits.

According to Mendelian inheritance patterns, the F2 progeny of male and females combined

would display a 9:3:3:1 phenotypic ratio of wild-type body and wild-type eyes, wild-type body

and cinnabar eyes, yellow body and wild-type eyes, and yellow body and cinnabar eyes,

respectively. The female flies’ phenotypic ratio would result in a phenotypic ratio of 6:2

consisting of wild-type body and wild-type eyes and wild-type body and cinnabar eyes. In

comparison, male flies’ would prove similar to this experiment’s results with a 3:1:1:3

phenotypic ratio of wild-type body and wild-type eyes, wild-type body and cinnabar eyes,

yellow body and cinnabar eyes, and yellow body and wild-type eyes respectively.

Typically, the yellow gene (y) produces black body pigmentation whereas the mutation

causes a yellow pigmentation. A functioning yellow gene is involved in processes underlying

melanization and male sexual behaviors. Although it’s known that the yellow gene is involved in

these processes, the exact biological mechanism continues to be investigated. Recent evidence

supports theories that yellow may operate similar to a hormone or growth factor. This claim is

supported by comparable characteristics between yellow and hormones, such as its resemblance

and biochemical secretion and binding (Drapeau, 2003). In contrast, a mutation in the cinnabar

gene (cn) results in a bright red eye phenotype. Operative cinnabar eye pigmentation encodes

products and enzymes necessary for producing brown eye pigmentation. Specifically, the

cinnabar gene encodes the third enzyme in the kynurenine 3-monooxygenase pathway, which is

located exclusively in developing eye tissue (Warren et al, 1996).

Ultimately, these experimental findings presented supported the three laws of Mendelian

inheritance: the law of dominance, the law of segregation, and the law of independent

assortment. In the dihybrid cross of Drosophila melanogaster between true-breeding virgin

yellow sex-linked mutant females and true-breeding cinnabar autosomal mutant males, F1 and

F2 progeny demonstrated evidence of the three laws and produced phenotypic ratios similar to

the expected 3:1:1:3.

Literature Cited

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Crosse: Biology 306 Genetics Laboratory Manual. University of Wisconsin-La Crosse in
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Drapeau, M.D. 2003. A novel hypothesis on the biochemical role of the Drosophila
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Trajkovic, J., Vujic. V., Milicic, D., Gojgic, G., Pavkovi, S, and Savic, T. 2017. Fitness traits of
Drosophila melanogaster (Diptera: Drosophilidae) after long-term laboratory rearing on
different diets. European Journal of Entomology. 114: 222-229.
Warren, W.D., Palmer, S., and Howells, A.J. 1996. Molecular characterization of the cinnabar
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