Sultan Qaboos University

College of Science
Department of Biology
BIOL4432Genetics
Spring 2012

Report of:

Drosophila
(White vs. bar)

Done by :
Munira said AL-yaroubi(85814)
Abir Omar Hilal AL-Rumhi (87142)

Introduction:
The fruit fly has been the subject of genetic studies since
about 1909.Drosophila melanogaster is the first
organisms to be studied genetically. As most species,
Drosophila melanogaster is diploid. It has four pairs of
1

chromosomes, three pairs are autosomal and one pairs is

sex chromosomes. Female drosophila is XX, and males
XY. The widely use of drosophila in genetic analysis refers
to several attributes, that this organism possess: small
size, high reproductive rate (400-500 in10days), short life
cycle (10-14 days at 25c), easily raised, cultured in the
laboratory, it shows huge variation of naturally occurring
and artificially induced genetic variants .In addition, the
large chromosome found in the salivary gland cells are
valuable for many types of chromosome studies.
Life cycle of drosophila
The flies spend five days in the egg and larval stages and
four days in the pupa stage. The adult flies may live for
several weeks. Drosophila cultures should not be exposed
to high temperatures (e.g. above 30o C) which result in
sterilization or death of the flies or to low temperatures
(e.g. below 10o C) which result in a prolonged life cycle
(perhaps 57 days) and reduced viability. The adult
Drosophila female starts to deposit eggs on the second
day after emergence. Embryonic development of the egg
takes about one day at 25 C. The larva is white,
segmented and worm like. The larval stage is a feeding
stage and consists of three subdivisions called instars.
The first and second instars stages end in molting which
allows the larva to grow. The third instar ends with
pupation. Prior to pupation, the animal stops feeding and
crawls to some relatively dry surface and the cuticle
hardens and darkens to form the puparium (like a
cocoon). Metamorphosis occurs in the puparium and
takes about four days. The pupa begins to darken just

prior to the emergence of the adult fly. Most flies emerge

from the pupa in the early morning hours.

Fig1: life cycle of drosophila

Background on white and bar mutation

White, abbreviated (w) was the first sex-linked mutation
ever discovered in the melanogaster. The white gene is
normally located on the X chromosome (chromosome
number 1) exactly at position 1.5 and the white allele is
recessive to the wild w+.Bar, abbreviated (B) was one of
the autosomal mutation, that is responsible for eyes
small, slithers or kidney shaped when heterozygote .The
bar gene is normally located on the X chromosome
(chromosome number 1) exactly at position 57.0 and bar
allele is dominant to the wild B+.

Analysis of our study

In our project we going to analysis the dihybrid cross
between white eyes female drosophila vs bar eyes males.
The symbols:
-Recessive =w, dominant = B
-white =w, wild =w+
+
-bar =B, wild= B
-white female: B+B+XWXW, bar male:
BBXW+XW+

Aims and objectives:

i.
ii.
iii.

Conduct an experiment using live organisms, a subset of the

population of Drosophila melanogaster.
Observe and understand the life cycle of drosophila.
To use reciprocal crosses to determine if an unknown Drosophila
mutation is dominant or recessive, and if it is autosomal or X-linked.
3

iv.
v.
vi.
vii.

Construct crosses of caught and known wild- type and mutated flies.
Learn techniques to manipulate flies and sex them.
Improve thinking skills and connection in the students.
Determine the inheritance pattern of a gene using a multigenerational

viii.

experiment.
Analyze data from your genetic crosses using chi-square analysis

techniques.
Materials and methods
i. Vial of bar eyes males and females of Drosophila
melanogaster with food at the bottom.
ii. Soft, small paint brush for moving etherized flies.
iii. Etherizer and ether (Or Fly nap kit).
iv. Empty vials.
v. Hand lens or Dissection microscope.
vi. Media formula 4-24 (available from Carolina biological
supply company).
vii. Small boats.
viii. Yeast.
Methods :
Preparing media;
i. Put 2 small boats of Media in empty vials.
ii. Power 2 small boat of tap water in media.
iii. Put 2 pieces of yeasts in media -water
mixtures.
Observation of Parental
i. Etherize the vial of wild-type flies.
ii. Using the dissecting scope, determine the
differences between sexes.
iii. Etherize one vial of mutant flies at a time.
iv. Using the dissecting scope, compare the
mutant traits to the wild-type traits.
v. Record the phenotype and genotype of the
flies.
vi. Repeat steps 3 through 5 with the second
vial.
Put the bar male eye
and female you have
separate in one vial
and the white in other

Day 1

vial and let them to

intermarry.
Remove parents from
cultures to prevent
breeding between
generations.

Day 5

The eggs of bar and

white hatched and you
get virgin flies.

Day 9

Put at least 4 virgin

females flies ( bar) of
one strain and at least 4
males ( white ) of the
other strain into one
culture vial .

Day11

Larvae of two
intermarriage strain
(bar / white ) emerge.
Remove parents from
cultures to prevent
breeding between
generations.

Day 12-20

Appearance of first
generation. Determine
the phenotype of
generation and Count
the number of
generations in each
trait.
Count the number of
females and males.
Cross males and
females of F1, 4 at least
from each sex.

Day 21-25

(7-10 days later).

Larvae of the
intercrossing F1 appear.
Remove the F1 adults

Day 30-35

from the culture.

Appearance of second
generation (F2) on the
day 35.

(after F2 generation
emerges)
Anesthetize and count
the flies every other
day for about 10 days.
Once counted, the
flies should not be
returned to the same
culture vial.

Day 36-47

Result:

Female
male

total

phenoty
pe
Red bar
eye
White
bar eye

F1
N0 perce
.
nt
34 52.31
%
31 47.69
%
65

100%

F2
N0.

phenoty
pe
wild/white eye

perce
nt
12%

Bar/whit

27

36%

29

38.67
%

wild/red eye

10

Total

75

13.33
%
100%

e eye
Bar/red
eye

DISCUSSION:
According to information about our mutation we
expected is that the F1 will not be wild type, because
autosomal sex linked alleles crossing
Will not be as autosomal autosomal alleles
crossing.therefore, F1 ratio 1:1 and F2 3:3:1:1.
As it is appeared from the above result in table2 that
show the observe result is that the percentages of F1
are 47.69% and 52.31% that are corresponding to 1:1
ratio.
While for the F2 are 36%, 38.67%,12% and 13.33% that
are corresponding to 3:3:1:1 ratio.
In fact, our finding result as show in table2 matched our
expected result in table 3 and 4.

F1
phenotype

B+B+XWXW
BB+XWXW+
Red wild/bar

BB+XWXW+
Red wild/bar

B+BXWY
white

BBXW+Y
B+BXWY
white

eye

eye

wild/bar eye

wild/bar eye

Wild= W+ (red eye), White=W

Bar= B (dominance), Wild= B+
F2 generation:

B+BXWXW+

B+BXW+Y

B+XW

B+XW+

BXW

BXW+

B+XW

B+Y

BXW

BY

B+B+XWXW

B+B+XWY

B+BXWXW

B+BXWY

wild/white eye

wild/white eye

Bar/white

Bar/white

eye

eye

B+B+XW+XW

B+B+XW+Y

B+BXW+XW

B+BXW+Y

Normal/red

Normal/r

Bar/red

eye

ed eye

eye

B+BXWXW

B+BXWY

BBXWXW

BBXWY

Bar/white

Bar/white

Bar/white

Bar/white

eye

eye

eye

eye

BB+XWXW+

B+BXW+Y

BBXW+XW

BBXW+Y

Bar/red

Bar/red

Bar/red

Bar/red eye

eye

eye

eye

Bar/red eye

Table 5: Chi Square Analysis.

Phenotype

Wild/whit
e eye
Wild/red
eye
Bar/white
eye
Bar/red
eye

#
Observed
(o)
9

#
Expected
(e)
9.375

(o-e)

-0.375

0.140625

0.015

10

9.375

0.625

0.390625

0.0417

27

28.125

-1.125

1.265625

0.045

29

28.125

0.875

0.765625

0.0272

(o-e)2

(o-e)2
e

Conclusion:
To conclude we have to keep in our mind many
considerations
To improve our result in this experiment, keeping fly
culture in fixed condition (tem. Humidity...etc.),
amount of the yeast must be two prices for two
media small boat and water and avoid moving with
fly culture everywhere and must be handled
carefully.
In addition we faced several problems such change
in temperature during the two last month which
affect the crossing process negatively and difficulty
to determine exactly the symbol of our mutation.
Finally this project allowed us to apply Mendelian
low of inheritance .