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TITLE : Drosophila melanogaster

OBJECTIVE :

i. To calculate the offspring characteristics that given same to the Mendel’s law of
inheritance.
ii. To make students creative to do the experimental design the crosses of flies.

INTRODUCTION:

Drosophila melanogaster is a small, common fly found near unripe and rotted fruit. It has
been in use for over a century to study genetics and lends itself well to behavioral studies.
Thomas Hunt Morgan was the preeminent biologist studying Drosophila early in the
1900's. Morgan was the first to discover sex-linkage and genetic recombination, which
placed the small fly in the forefront of genetic research. Due to it's small size, ease of
culture and short generation time, geneticists have been using Drosophila ever since. It is
one of the few organisms whose entire genome is known and many genes have been
identified.

Fruit flies are easily obtained from the wild and most biological science companies carry
a variety of different mutations. In addition these companies sell any equipment needed
to culture the flies. Costs are relatively low and most equipment can be used year after
year. There are a variety of laboratory exercises one could purchase, although the
necessity to do so is questionable.

PROCEDURE:

There are a few methods in fainting the flies. In this practical, we are given chloroform of
anesthetization kit for fainting the flies. The lab assistants show to us the technique. The
newly fainted flies should be examined immediately. Always ready with cotton or cotton
buds with the anesthetizer before hands as to avoid the sudden awaken of the flies.

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Part I
i. Firstly we identify the wild type and mutant flies. We had examined their
morphology thoroughly before we do the crosses. We use the dissecting
microscope in lab to examine the flies.
ii. We had design our experiment properly in order to yield expected results.
iii. We choose the traits that can breed through based on autosomal and sex-linked.
iv. Before we hands the flies, we show our experimental design to our Tutor for
approval. Then we inform the lab assistant the flies that we want to use.

Part II

i. We have chosen five pairs of traits that follow: Mendel Law 1, Mendel Law 2 and
X-linked.
ii. We had bred the homozygote parents of wild type and homozygote mutant.
iii. When we see the larvas have emerged from the medium, we took out the parents
and kill them immediately. The Tutor and lab assistants had shown us how to kill
the flies. We also make sure that no one of flies flying in the lab.
iv. We let the larvas grow. We placed the virgin females in new tubes with their male
siblings. When we see another generation of larvas (F2) emerge, we killed the F1.
v. Every day we go to lab to see if we get the F 2 generation or not. If we have that
flies, we anesthetize the adult F2 and we count.
vi. We had analysed the results using Chi-square.

Parents generation
Mendel Law 1 Vestigial Wing x Wild Type
Mendel Law 1 Ebony x Wild Type
Mendel Law 2 Sephia, Ebony x Wild Type
X-linked White Eye (M) x Wild Type (F)
X-linked Wild Type (M) x White Eye (F)

RESULT & DISCUSSION:

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Mendel Law 1

Law 1 : Vestigial Wing x Wild Type

Legend : S- Dominant allele control the wild type wing trait


s- Recessive allele control the vestigial wing trait

Cross : VV x vv

Result (f2 generation): 3:1

Characteristic O (observation E (Expected values) (O-E)2


values)
Vestigial 21 1/4 x 130 = 32 121
Wild Type 109 ¾ x 130 = 98 121

CHI-SQUARE TEST:
Degrees of freedom = 2-1
=1

X2 = ∑ (O-E)2 / E

Where,
O = Observed data in each category
E = Observed data in each category based on the experimenter’s hypothesis
S = Sum of the calculation for each category

X2 = ∑ (21-32)2/ 33 + (109-98)2/ 98

= 4.9

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= 1%< X2 <5%

= 0.01< X2 < 0.05

Discussion:

From the result, the observation value is 21 for vestigial wing and 109 for wild type wing.
So, wild type is dominant to vestigial wing. Based on the observations, the hypothesis is

‘Wild type (V) is dominant to the Vestigial (v).

From the chi-square test, the chi-square value is lower than 0.05. So, that means the null
hypothesis is rejected for the experiment. From the chi-square test the data and
hypothesis do not have a good fit.

During the experiment, the data are collected not accurately due to the lost of the
Drosophila melanogaster during the researcher change the place of the Drosophila
melanogaster and the another factor is due to the fatal problem.

Mendel Law 1

Legend:
E- Dominant allele for wild type
e- Recessive allele for ebony body

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Parent : Ebony x Wild type
ee EE

G1 :
e E

F1 : All wild type


Ee

F1xF1 : 5 wild type x 5 Wild type


Ee Ee

G2 :
E e E e

38 : 119 x2 = ∑(O-E)2/ E
F2 :
Ebony
M/F EWild type e
x2 = Chi square value
E Ee Ee O = Observed data in each
E Ee ee category
E = Data in each category based
Chi square test: x2 = ∑(O-E)2/ E on the experiments
hypothesis

Drosophilla types Observed data (O) Experiments’ (O-E)2/ E


hypothesis (E)
Wild type 119 ¾ x 157 = 117.75 (119-117.75) /
117.75 = 0.013
Ebony 38 ¼ x 157 = 39.25 (38-39.25) /
39.25 = 0.040
Total 157 x2 = 0.053

Discussion:

Mendel Law 1, or Law of Segregation states that, the two copies of a gene segregate from
each other during transmission from parent to offspring. This law predicts that the
phenotypes of the F2 generation will be in ratio of 3:1 of dominant to recessive traits.

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Therefore, for this experiment, the hypothesis of Mendels Law 1 is said to be 3:1 ratio
of wild type to ebony type of Drosophilla melanogaster.

From the cross between F1 generation, they produce two types of Drosophilla offsprings
which are ebony type and wild type. Thus n= 2. As degree of freedom, df= n-1, therefore
df value is 1.

With the df=1, and the Chi Square value equal to 0.053, we have obtained results that
near to 0.0642 value under probability of 0.80.

Therefore from the table of Chi Square values and Probability, the obtained chi square
value from experiment which is 0.053, are expected to occur 80% of the time based on
random chance alone when hypothesis are correct. This low value of chi square indicates
that the observed value that we obtained from the experiments is actually fit the
hypothesis. Thus, our result can be well accecpted. The results obtained from the
experiments fits the hypothesis value of Mendels law 1. The gene separate from each
other during transmission from parent to offspring and cause the cross of F1 to F1
producing F2 offspring that obey Mendels law 1, with 3:1 ratio of wild type to ebony
type of Drosophilla melanogaster.

Mendel Law 2
Mendel stated for the Law 2, two different genes will randomly assort their alleles during
the formation of haploid cells. The expected ratio is usually 9:3:3:1

Parent phenotype: Sephia, ebony x wild type

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(Female) (Male)

From the cross above, the predicted outcome is sephia ebony, sephia wild type, red ebony
and wildtype.

Sephia ebony Sephia wildtype Red ebony Wildtype


110 50 49 12

Total 221

The ratio get from this data is 9:3:3:1

To test the validity:


Using the Chi-Square Test
Degrees of freedom = 4-1
=3

X2 = ∑ (O-E)2 / E

Where,
O = Observed data in each category
E = Observed data in each category based on the experimenter’s hypothesis
S = Sum of the calculation for each category

Through Chi- Square Test:

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Characteristics Observed Observed data based on x² = (O-
data (O) experimenter’s hypothesis ( E) E)²
E

Sephia ebony 110 9/16 x 221 = 124.3 1.645

Sephia wildtype 50 3/16 x 221 = 41.4 1.786

Red ebony 49 3/16 x 221 = 41.4 1.395

Wildtype 12 1/16 x 221 = 14


0.286
Total 221 5.112

Discussion:

At first, we have known that df = 3. Thus, from the chi square the value obtained
is 5.112. From the table of chi square this value is greater than 4.642 and less than 7.815.
So from that we know that P value is located between 0.05 and 0.20 or 5% to 20%. Since
the value is between 5% and 20%, and not exceed 0.05, this value is accepted and the
data and the hypothesis have a good fit. From the cross, the number of sephia ebony is
higher than others and the wildtype has the lower number.

X- linked

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Parent phenotype white eye x wild type
(Female) (Male)

F 2 phenotype
Male Female
Red eye 25 45
White eye 50 41

Characteristics Observed data (O) Observed data x² = (O- E) ²


based on E
experimenter’s
hypothesis ( E)

Red eye male 30 ¼ x 166 = 41.5 3.187

Red eye female 45 ¼ x 166 = 41.5 0.295

White eye male 50 ¼ x 166 = 41.5 1.741

White eye female 41 ¼ x 166 = 41.5 0.006

Total 166 5.229

Discussion:

With df = 3, the chi square value of 5.229 is slightly greater than 4.642 and less
than 7.815 which give a P value between 0.05 and 0.20 or 5% to 20%. Since the value is
between 5% and 20%, based on random chance alone, thus the hypothesis is accepted.
The data and the hypothesis have good fit even though the P value is quite low.

X – Linked (Parent: White – eyed male to wild – type female)

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Female (F=38) Male (M=33)
Red eye White eye Red eye White eye

36 2 12 21

Discussion:

Based on our result in the table above, the experiment is done successfully. To get
this result, we have crossed the white – eyed male to a wild – type or true – breeding red
– eyed female. The result for the offspring or the F1 generation has been recorded. All of
the F1 offspring had red eyes due to red is dominant compare to white. Then, F1 offspring
were mated to each other to obtain an F2 generation.

After we crossed the F1 offspring with each other, the result is recorded just like
in the table above. We obtained 38 female and 33 male. We got 36 red – eyed female and
2 white – eyed female while 12 red – eyed male and 21 white – eyed male.

If we compare to the experiment that have been done by Thomas Hunt Morgan
who is confirmed about the chromosome theory of inheritance, there is different is some
part of the same experiment. He also obtained that all of the F 1 offspring had red eyes
following X-link cross ratio of F1 which all progeny should be red. It same goes to us.
But the different is F2 generation. He most notably that no white – eyed female offspring
were observed in the F2 generation. According to expected theory we should never get the
female F2 progeny with white eye. So, in this experiment, we might be late in observing
the F2 progeny as we conclude that the 2 white eye female of F2 progeny is actually not
F2 generation but the F3 generation. We also conclude that, even if the 2 white eye
female are from F2 generation, the dominant allele of red eye in the X chromosome

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undergo mutation, so, it did not show the characteristic of dominant allele and the
recessive allele on the other X chromosome becomes more dominant than the mutated
allele. Hence, both Drosophila melanogaster shows recessive appearance. It suggested
that the pattern of transmission from parent to offspring depend on the sex of the
offspring and on the alleles that they carry. That’s why he concluded that the eye color
alleles are located on the X chromosome because the prediction was confirmed
experimentally.

For our result, it doesn’t follow the Mendel’s Law or Morgan’s proposal because
we obtained the white – eyed female. But our experimental just undergo one time
compared to Morgan’s experiments. So, might be there is error that occur during we were
mating the fruit flies or also known as Drosophila. But, the number of the white – eyed
female is only 2. So, it very closes to the theory.

CONCLUSION:

As the conclusion, the objective of this experiment is accepted where the students have
knowledge and creative to design the experiment of crosses the flies. We also can
identify the characteristics of Drosophila melanogaster using the dissecting microscope.

REFERENCE:
Robert J. Broker ( 2005 ). Genetics analysis & Principles. McGraw Hill International
Edition.

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APPENDIX:

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Pictures of Drosophila melanogaster

White eye Vestigial wing

wildtype Red eye

Red ebony Sephia ebony

White eye

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