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GENETICS
NAME
D20091034843
AMEERA BT YAHYA
D20091034814
D20091034858
: 2 AUGUST 2010
The Law of Independent Assortment, also known as "Inheritance Law", states that
alleles of different genes assort independently of one another during gamete
formation. While Mendel's experiments with mixing one trait always resulted in a 3:1
ratio between dominant and recessive phenotypes, his experiments with mixing two
traits (dihybrid cross) showed 9:3:3:1 ratios. Mendel concluded that different traits are
inherited independently of each other, so that there is no relation, for example,
between a cat's colour and tail length. This is actually only true for genes that are
not linked to each other.
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 its 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, 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. In this experiment Drosophila are use because they are small and easily
handled, Drosophila are sexually dimorphic (males and females are different), making it is
quite easy to differentiate the sexes, and flies have a short generation time (10-12 days) and
do well at room temperature.
Life cycle of Drosophila melanogaster
D. melanogaster exhibits complete metamorphism, meaning the life cycle includes an egg,
larval (worm-like) form, pupa and finally emergence (eclosure) as a flying adult. This is the
same as the well-known metamorphosis of butterflies and many other insects. The larval
stage has three instars, or molts.
Life cycle by day
Day 0: Female lays eggs
Day 1: Eggs hatch
Day 2: First instar (one day in length)
Day 3: Second instar (one day in length)
Sex difference
Several criteria may be used to distinguish male and female Drosophila melanogaster
Abdomen shape the female abdomen curve to a point, the male abdomen is round
and much shorter. Figure below show Male (left) and Female (right) wild-type
Drosophila.
Mark on their abdomen - Alternating dark and light bands can be seen on the entire
rear portion of the female; the last few segments of the male are fused.
Sex comb - On males there is a tiny tuft of hairs on the basal tarsal segment of the
front leg. This is the only sure method of distinguishing young male and female flies
(less than 2 hours old), since the other adult traits are not always immediately
recognizable. Sexing via the sex comb can also be done successfully in the pupal
stage (Hadden and Cunningham, 1970).
Sex organ at abdomen - The genitalia are the easiest and most reliable character to
use in determining sex (right; ventral view, posterior is up). Note the circle of darkly
pigmented parts in the male. In contrast, the tip of the female's abdomen is lightly
colored and pointed.
Dorsal view
Hereditary Traits
Before one observes their mutants, one needs to be familiar with the appearance of the wild
type Drosophila, the type found most often in natural populations of the organisms. Although
thousands of mutations in Drosophila are known, only those which are relevant to these
exercises are listed.
1. Eyes
Wild type: red, oval in shape and many-faceted
Mutants: white, black, apricot, scarlet red, pink, or brown; changes in shape and number of
facets
2. Wings
Wild type: smooth edges, uniform venation, extend beyond the abdomen
Mutants: changes in size and shape; absence of specific veins; changes in position in which
wings are held when at rest
3. Bristles
Wild type: fairly long and smooth (note distribution on head and thorax)
Mutants: shortened, thickened, or deformed bristles changes in patterns of distribution
4. Body colour
Wild type: basically gray, with pattern of light and dark areas
Mutants: black (in varying degrees), yellow, in doubtful cases, color can often be determined
most clearly on wing veins and legs
Mutants traits can be assumed as recessive to the wild type.
and strike slowly until the adult Drosophila drop down in to the bottom.
Quickly, the bottle is separated from its cover.
The flies are then subjected to the ether for a minute or until they ceased moving.
After that, the etherized flies are transferred on the filter paper.
If the flies revived before we finished examining them, a few drops of ether is added
to the cotton and put in the petri dish and covered it.
8. The etherized flies are examined with a dissecting microscope.
9. A soft brush is used for moving the flies about on stage of the microscope.
10. Finally, after finishing our experiment, the Drosophila is discarded in a soup water or
mineral oil, except the ones we need for the further crosses which we have to put on
dry surface in the culture bottle before they come into contact with the moist medium.
Procedure in the experiment
The wild type and mutant flies are identified. Their morphology are examined
thoroughly before we do the crosses. We will use dissecting microscope. Here are
some traits for the experiments:
Mendel Law 1
- Ebony body
- curved wings
- Sepia eyes
- Scarlet eyes
Mendel Law 2
- Ebony body, yellow body
X-linked
- white eyes
- yellow body
- echinus eyes
- bar eyes
Procedures for monohybrid crosses
1.
In monohybrid crosses, red eyes drosophila (male) and scarlet eyes (female )
2.
3.
4.
ANALYSIS OF F2 GENERATION
1.
By using the ether/ Flynap, the F2 Drosophila is killed and is put on the filter
2.
paper.
The total number of every F2 phenotype is counted. From the monohybrid
crosses, there are TWO phenotypes only. The distribution of the F2
4.
In dihybrid crosses, wild type drosophila (male) and vestigial, sepia eyes
2.
3.
After a few days, the Drosophila will mate and finally the female Drosophila
will lay eggs which then will hatch. At this moment, all the parental Drosophila
4.
ANALYSIS OF F2 GENERATION
1.
By using the ether/ Flynap, the F2 Drosophila is killed and is put on the filter
2.
paper.
The total number of every F2 phenotype is counted. From the dihybrid
crosses, there are FOUR phenotypes.The distribution of the F2 phenotypes is
4.
3.
4.
Results
Monohybrid Crosses
st
+
st
Gamete
F1
st+st
All wild-type
F1 F1
st+st
st+st
Gamete
st
st+st+
+
F2
Ratio
st
st+st
st
st+st
+
st
stst
3: 1
wild-type : scarlet-eye
Number of progeny
Females
23
0
Males
13
0
Wild type
Scarlet eyes
Total
36
0
The further whether eye color was inherited according to Mendelian laws, we crossed
the F1 progeny and examined the phenotypes of the resulting F2 flies.
Phenotypes
Number of progeny
Females
38
12
Males
27
7
Wild type
Scarlet eyes
Total
65
19
84
To determine the statistical relevance of the data, we performed the Chi square test
on our F2 data.
Class
Wild type
Observed
65
Scarlet eyes
Totals
19
84
Expected
63
(O-E)2
( 65-63)2
(O-E)2/ Expected
4/ 63
21
=4
( 19-21)2
=0.06
4/21
=4
=0.19
X2 = 0.25
84
Gamete
st
+
F1
st
st+st
All wild-type
F1 F1
st+st
st+st
Gamete
F2
st
+
st
st
+
st+st+
st+st
st+st
st
stst
Ratio
3: 1
wild-type : scarlet-eyed
To test Mendels Law of Segregation, we examined the inheritance of eye color by
crossing two pure breeding strains of Drosophila melanogaster that is wild type and
scarlet eyes. We determined which allele is dominant by setting up the cross st+st+
males stst females as described above. The phenotypes of the progeny are shown
below.
Phenotypes
Number of progeny
Males
13
0
Wild type
Scarlet eyes
Females
23
0
Total
36
0
The further whether eye color was inherited according to Mendelian laws, we crossed the F 1
progeny and examined the phenotypes of the resulting F2 flies.
Phenotypes
Number of progeny
Females
32
9
Males
21
6
Wild type
Scarlet eyes
Total
53
15
68
To determine the statistical relevance of the data, we performed the Chi square test on our
F2 data.
Class
Wild type
Scarlet eyes
Totals
Observed
53
Expected
51
15
17
68
The degree of freedom, df
(O-E)2
(O-E)2/ Expected
(53 51)2
4/51
=4
(15 17)2
= 0.08
4/17
=4
= 0.24
68
X2 = 0.32
= n-1 (n= total number of categories)
= 2-1
=1
from the cross. The result of the parental cross ( st+st+ females x stst male ) show the same
result parental cross ( st+st+ males stst females)
CONCLUSION
In conclusion, the phenotype of the F1 progeny confirmed that the allele for wild type scarlet
eyes, st+ is dominant to the allele for scarlet eyes, st. The ratio of normal-eyed (wild-type) to
scarlet-eyed flies of 3.42:1 seen in the F2 is very near that of the expected 3:1 ratio for a
monohybrid cross, and the Chi square test verifies that it is within statistical limits. Therefore,
the results of this experiment confirm Mendels Law of Segregation. The result of the
parental cross ( st+st+ females x stst male ) show the same result parental cross ( st+st+
males stst females) because scarlet eye does not located on sex chromosome.
Dihybrid Crosses
Dihybrid crosses involve manipulation and analysis of two traits controlled by pairs of alleles
at different loci.
In this experiment, we cross wild type with vestigial wing, sepia eyes
e is recessive allele for sepia eyes
e+ is dominant allele for wild-type eyes
vg is recessive allele for vestigial wing shape
vg+ is dominant allele for wild-type wing shape
Vestigial male
vestigial female
The cross between wild type male and vestigial, sepia female
Wild type male
e+e+ vg+vg+
Parent
Gamete
ee vgvg
e+
vg+
e vg
e+e vg+vg
F1
All wild-type
e+e vg+vg
F1 F1
e+vg+
e+vg
e+e vg+vg
evg+
ev
g
e+vg+
e+vg
evg+
ev
g
e vg
e+ vg
e vg+
e vg
e+ vg+
e+e+ vg+vg+
e+e+ vg+vg
e+e vg+vg+
e+e vg+vg
e+ vg
e+e+ vg+vg
e+e+ vgvg
e+e vg+vg
e+e vgvg
e vg+
e+e vg+vg+
e+e vg+vg
ee vg+vg+
ee vg+vg
e vg
e e vg vg
e+e vgvg
ee vg+vg
ee vgvg
+
expected genotypic and phenotypic ratios from an analysis of these 16 possibilities can be
calculated.
To test Mendels Law of Independent Assortment, we examined the inheritance of eyes
colour and wing shape by crossing two pure breeding strains of Drosophila melanogaster
that is wild-type and vestigial, sepia. We determined which allele is dominant by setting up
the cross e+e+ vg+vg+ males ee vgvg females as described above. The phenotypes of the
progeny are shown below.
Phenotypes
Wild type
Vestigial, sepia
Males
Number of progeny
Females
Total
9
0
13
0
22
0
The further whether eyes colour and wing shape was inherited according to Mendelian laws,
we crossed the F1 progeny and examined the phenotypes of the resulting F2 flies.
Phenotypes
Number of progeny
Females
34
9
7
4
Males
23
5
4
2
wild type
sepia
vestigial
sepia, vestigial
Total
57
14
11
6
88
To determine the statistical relevance of the data, we performed the Chi square test on our
F2 data.
Class
wild type
sepia
vestigial
sepia, vestigial
Totals
Observed
57
Expected
9/16 88
(O-E)2
(57 - 50)2
(O-E)2/ Expected
49/50
14
= 50
3/16 88
= 49
(14- 16)2
= 0.98
4/16
11
= 16
3/16 88
=4
(11 16)2
= 0.25
25/16
= 16
1/16 88
= 25
(9 6)2
= 1.56
9/6
=6
=9
= 1.50
X2 = 4.29
88
88
The cross between wild type female and vestigial, sepia male
Parent
Gamete
e+e+ vg+vg+
ee vgvg
e+
vg+
e vg
e+e vg+vg
F1
All wild-type
e+e vg+vg
F1 F1
e+vg+
e+vg
e+e vg+vg
evg+
ev
g
e+vg+
e+vg
evg+
e vg
e+ vg+
e e vg+vg+
+ +
e+ vg
e e vg+vg
+ +
e vg+
e e vg+vg+
+
e vg
e+e vg+vg
ev
g
e+ vg
e vg+
e vg
e+e+ vg+vg
e+e vg+vg+
e+e vg+vg
e+e+ vgvg
e+e vg+vg
e+e vgvg
e+e vg+vg
ee vg+vg+
ee vg+vg
e+e vgvg
ee vg+vg
ee vgvg
In a dihybrid cross, each of the F1 parents can produce four different gamete types, so there
are 16 (= 4 x 4) possible offspring combinations. Because the two traits show complete
dominance and separate independently of each other (Law of Independent Assortment), the
expected genotypic and phenotypic ratios from an analysis of these 16 possibilities can be
calculated.
To test Mendels Law of Independent Assortment, we examined the inheritance of eyes
colour and wing shape by crossing two pure breeding strains of Drosophila melanogaster
that is wild-type and vestigial, sepia. We determined which allele is dominant by setting up
the cross e+e+ vg+vg+ males ee vgvg females as described above. The phenotypes of the
progeny are shown below.
Phenotypes
Number of progeny
Females
21
0
Males
13
0
Wild type
Vestigial, sepia
Total
34
0
The further whether eyes colour and wing shape was inherited according to Mendelian laws,
we crossed the F1 progeny and examined the phenotypes of the resulting F2 flies.
Phenotypes
Number of progeny
Females
33
9
10
4
Males
23
6
6
2
wild type
sepia
vestigial
sepia, vestigial
Total
56
15
16
6
93
To determine the statistical relevance of the data, we performed the Chi square test on our
F2 data.
Class
wild type
sepia
Observed
56
Expected
9/16 93
(O-E)2
(56 - 53)2
(O-E)2/ Expected
9/53
15
= 53
3/16 93
=9
(14- 17)2
= 0.17
9/17
Vestigial
sepia, vestigial
Totals
16
= 17
3/16 93
=9
(16 17)2
= 0.53
1/17
= 17
1/16 93
=1
(6 6)2
= 0.06
0/6
=6
=0
=0
X2 = 0.76
93
93
CONCLUSION
In conclusion, the phenotype of the F1 progeny confirmed that the allele for wild type, e+e+
vg+vg+ is dominant to the allele for sepia, vestigial, ee vgvg. The ratio of wild type to sepia to
vestigial and to sepia, vestigial of 9.5:2.3:1.8:1 seen in the F2 is very near that of the
expected 9:3:3:1 ratio for a dihybrid cross, and the Chi square test verifies that it is within
statistical limits. Therefore, the results of this experiment confirm Mendels Law of
Independent Assortment. The result of the parental cross
vestigial, sepia female show the same result parental cross between wild type female and
chromosome.
Sex linked Traits
In sex-linked inheritance, alleles on sex chromosomes are inherited in predictable patterns.
In Drosophila the locus for eye color is located on the X chromosome. The allele for red eye
color, which is normal in wild flies, is dominant to the mutant allele for white eyes.
In the left hand example, homozygous red eyed females (RR) mate with homozygous white
eyed males (w-). In the offspring, all the daughters are red eyed heterozygotes (Rr) and all
sons are red eyed homozygotes (R-). In the right hand, homozygous white eyed females (rr)
mate with homozygous red eyed males (R-). In the offspring, all the daughters are red eyed
heterozygotes (Rr) and all sons are white eyed homozygotes (r-).
Females have two chromosomes X (with a locus for eye color), they might be homozygous
or heterozygous for either allele. Males, carry only one X chromosome, are always
hemizygous. They carry only the one X chromosome inherited from their mother, and it
determines their eye color.
The first cross is between normal female with red eyes and the mutant male with white eyes.
The first cross is to determine whether the white or red eyes were dominant. The F 1
generation all had red eyes. So we can conclude that red eyes are dominant over white.
Then, F1 generations are crossed each other to give F2 progenies.
From the result of the experiment, only male Drosophila shows white eyes. X-linked inherited
diseases occur far more frequently in males because they only have one X chromosome.
Females must receive a copy of the gene from both parents to have such a recessive
disease. However, they will still be carriers if they receive one copy of the gene.
Cross between red eye (wild type) female X white eye male
B+ Dominant allele codes for red eye
B Recessive allele codes for white-eye
Parental:
Genotype:
XB+ XB+
Gamete:
F1:
XB
XB
XB+XB
XB
XB+Y
XB+XB
XB+Y
Gamete:
F2:
XB+XB
XB
XB
XB+XB+
Phenotypic ratio:
XB+Y
XB
XB+Y
XB+XB XBY
white-eye (female)
Genotype:
XB XB
Gamete:
XB
F1:
XB
XB
XB+XB
XB Y
XB+XB
XBY
Gamete:
F2:
XB+XB
XB
XB
XB
XB+Y
XBXB
XB+XB
Phenotypic ratio:
male
XBY
XBY
RESULT
The female with red eyes will be crossed with white-eyed male to produce F 1 progeny. Then,
this F1 were crossed each other
R is dominant allele for wild type eyes (red eyes)
w is recessive allele for white eyes
The phenotype of F1 progeny is:
Phenotypes
Number of progeny
Males
Females
Total
Wild type
17
White eyes
Number of progeny
Males
Females
Total
Wild type
14
33
47
White eyes
16
16
Class
wild type male
Observed
14
Expected
1/4 63
(O-E)2
(14-16)2
(O-E)2/ Expected
4/16
Wild type
33
= 16
2/4 63
=4
(33-31)2
= 0.25
4/31
female
White eyes
16
= 31
1/4 63
=4
(16-16)2
= 0.13
0/16
male
White eyes
= 16
0/4 62
=0
(0 6)2
=0
36/6
female
Totals
=6
= 36
63
=6
X2 = 6.38
63
df = 3
With df = 3, the chi square value of 6.38 is slightly greater than 4.642 (which correspond to
P > 0.20). Therefore P = 0.20 show that expected to occur are 20% of the time. 80% error
occurred. Hypothesis accepted.
The male with red eyes will be crossed with white-eyed female to produce F 1 progeny. Then,
this F1 were crossed each other
R is dominant allele for wild type eyes (red eyes)
w is recessive allele for white eyes
The phenotype of F1 progeny is:
Phenotypes
Number of progeny
Males
Females
Total
Wild type
13
13
White eyes
15
15
Number of progeny
Males
Females
Total
Wild type
14
15
29
White eyes
13
14
27
Class
wild type male
Observed
14
Expected
1/4 56
(O-E)2
(14-14)2
(O-E)2/ Expected
0/14
Wild type
15
= 14
1/4 56
=0
(15- 14)2
=0
1/14
female
White eyes
13
= 14
1/4 56
=1
(13-14)2
= 0.07
1/14
male
White eyes
14
= 14
1/4 56
=1
(14-14)2
= 0.07
0/14
female
Totals
= 14
29
=0
29
=0
X2 = 0.14
markings on abdomen, appearance of sex comb external genitilia on abdomen and sex
organ during larval stage. This making easier for us to differentiate them especially in the
experiment about sex-linkage. What is more important is we are able to recognize wild-type
flies and those with classic mutations. To test the result that we obtained, we use the Chi
square statistic to test hypotheses concerning expected and observed ratios. If the p value is
less than 0.005, so the hypothesis can be rejected.
References
Paul Arnold (2009). Human Genetics and the Fruit Fly Drosophila Melanogaster.
Retrieved March 29, 2010, from http://www.biol.org/DrosPics.htm#Misc
Christin E. Arnini (1996). Using Drosophila to Teach Genetics. Retrieved March 29,
2010,http://www.google.com.my/search?
hlen&qdrosophila+melanogaster+phenotypes&revid.
Basic Genetics: Thomas Hunt Morgan and Sex-linked Traits. Retrieved April 6, 2010
from
http://library.thinkquest.org/20465/sexlinked.html
Miko, I. (2008) Thomas Hunt Morgan and sex linkage. Nature Education 1(1). Retrieved April
6,
and-Sex-Linkage-452#TB_inline?height=300&width=400&inlineId=trOutLine
Retrieve
on
April
2010
http://www.mun.ca/biology/dinnes/B2250/DrosophilaGenetics.PDF
Retrieved on 8 April 2010 at http://www.dreamessays.com/customessays/Science
%20Research%20Papers/11452.htm
at