UDBB 2233

Title : Exp 1-Chi Square Analysis

Exp 2- Mendelian Genetics of Drosophila and Maize

Name : Mohd. Zulfadhli B. Azizan 1004656

Lau Wai Yip 1004923

Siow Zi Kang 1002558

Cheah Jun Zhe 0905802

Ang Yee Von 1007253

Chan Jo Yee 0908033

Group : Practical 1

Lecturer : Mr. Kam Yew Chee

Experiment 1

Title: Chi-Square Analysis

Introduction:

The extent an observed data set fits or differs from the predicted or expected
occurrences can be evaluated by testing the goodness of fit of the data. When the data do
not fit exactly, we will want to find out how much deviation can be allowed before we
reject a null hypothesis. One of the simplest statistical tests to assess the goodness of fit
of the null hypothesis is chi-square analysis (x2), which tests the difference between
observed (O) and expected (E) values. This x2 value is then used to estimate how
frequently the observed deviation can be expected to occur strictly as a result of chance,
allowing us to determine whether progeny phenotype ratios fit our assumptions about
their genotypes. The formula for calculating the chi-square value is:
∑(O−E)2
x2= 𝐸
where O = the observed number of individuals in a particular phenotype,
E = the expected number in that phenotype, and
∑= the summation of all possible values of (O-E)2/E for the various phenotypic
categories

The degree of freedom (df), which is equal to n – 1, where n is the number of

categories (the number of phenotypes considered), will then be determined. Degrees of
freedom must be taken into account because the greater the number of categories, the
more deviation is expected by chance alone. The calculated _2 value can now be
interpreted at a corresponding probability value (p), which can be obtained from _2 table
(Appendix) for (n–1) degree of freedom.

The p value can be thought as a percentage. For example, a p value of 0.26

indicates that were the same experiment repeated many times, 26% of the trials would be
expected to exhibit chance deviation as great as or greater than that seen in the initial
trial. Conversely, 74% of the repeats would show less deviation than initially observed as
a result of chance. The interpretation of the p value reveals that a hypothesis is never
proved or disproved absolutely. Instead, a relative standard must be set to serve as the
basis for either rejecting or failing to reject the hypothesis. This standard is often a
probability value of 0.05. If the calculated _2 value is smaller than the critical value given
in the table at (n–1) df, then we accept the null hypothesis and conclude that the
difference between observed and expected results is not significant, and vice versa.
Objectives:

To 1.Calculate x2 to determine whether a data set approximates a theoretically expected

ratio.
2.Interpret a calculated x2 value at an appropriate number of degrees of freedom.

Materials:

Non-biological materials: container of equal quantities of colored and white beads, petri
dishes, two equal-value coins and a calculator.

Procedure :

Activity 1:
A beaker contained 150 colored and 150 white beads mixed were given.
1. One petri dish-full random sample of beads was removed from the beaker.
2. The beads of the different colors were segregated and counted.
3. The data was recorded in table 1.1. The expected numbers were calculated based on the
size of the sample and the known ratio of colored to white beads in the entire population.
The table 1.1 was completed and x2 was calculated.

Activity 2:
1. Two equal-value coins were together tossed 30 times.
2. The data were recorded in table 1.2 and the expected numbers were calculated. The
probability of two or more independent events occurring simultaneously is the product of
their individual probabilities were noted.
3. The table 1.2 were completed and x2 were calculated.

Activity 3:
A beaker contained 100 beads of two different colors in the ratio of 3:1 . The beads ere
thoroughly mixed.
1. One tablespoon-full random sample of beads were removed from the beaker.
2. The beads of different colors were segregated and counted.
3. The data in were recorded in table 1.3 and then the expected numbers based on the size of
the sample were calculated and the known ratio of colored to white beads in the entire
population. The table 1.3 were completed and x2 were calculated.

Activity 4:
The following are the approximate frequencies of the various ABO blood groups in a
hypothetical population: 41% A, 9% B, 3% AB, and 47% O. The number of students in the class
who have blood group A, B, AB and O were recorded in table 1.4 . The x2 were calculated to 2
decimal places and completed in table 1.4 .
Activity 5:
A container contained several thousand beads of 4 different colors in the ratio 9:3:3:1 were given.
The beads were thoroughly mixed.
1. One spatula-full random sample of beads were removed from the container.
2. The beads of different colors were segregated and counted.
3. The data were recorded in table 1.5 and then x2 were calculated.

Experiment 2
Title: Mendelian Genetics of Drosophila and Maize

(A) Drosophila Genetics

Introduction:
The vinegar or fruit fly, Drosophila melanogaster, has been widely used for
demonstrating the classical Mendelian laws. This small fly undergoes a complete
metamorphosis from egg, larva, pupa, through imago in 10 to 14 days at 25oC. The
fertilized eggs normally hatch into white larvae in 22–24 hours after they have been laid.
The larvae burrow in the culture medium and feed on it for about four days. They then
pupate on a dry spot, usually at the sides of the container. The pupation lasts for another
four to seven days before the adults emerge from the pupa case. The newly emerged
adults are pale in colour with unexpanded wings, but mature in about 8–10 hours with a
characteristic darkened body. The females usually do not mate before 8 hours after
emergence and do not lay eggs until they are two days old. In addition to a short life
cycle, Drosophila is highly prolific, and has a lot of genetic variability with many
morphological variants. Names and symbols are used to identify the mutant genes carried
by the flies. Below are some commonly used notations in Drosophila genetics.

a. Wild-type allele of any locus is indicated by the plus (+) symbol.

b. Mutant alleles are shown with italicized letters. A capital letter designates a dominant
gene, the lowercase letter represents recessive allele. E.g.,
B represents the dominant ‘Bar’ gene;
Bd represents the dominant “Beaded’ gene;
b designates the recessive ‘black’ gene;
bt designates the recessive ‘bent’ gene.

To represent a wild-type locus, a superscript + symbol is used after the letter. E.g.,

b+ shows that the wild-type gene is dominant over its recessive black gene, b;
B+ indicates that the wild-type gene is recessive to its dominant ‘Bar’ gene, B.
c. If more than two alleles are observed for a gene locus, mutant alleles can be
distinguished by small letters in superscript after the alphabet for the genes. E.g.,

w+ represents a wild-type eye-colour gene;

w represents a mutant white-eye gene;
wa represents a mutant apricot-coloured eye gene

d. The genotype of a gene locus for a diploid organism is represented by two symbols. E.g.

homozygote (wild-type) b+/b+;

homozygote (black mutant) b/b;
heterozygote (wild-type) b+/b.

e. To show two unlinked loci in a diploid, separate the two pairs of symbols. E.g.

b/b, se/se (black and sepia)

f. If two loci are linked, they are indicated by a slash. E.g.,

b vg/b vg (black and vestigial, coupling phase);
b vg+/b+ vg (phenotype: wild-type, repulsion phase).

g. Y chromosome is shown as Y, e.g.,

White-eye, miniature winged male can be written as w m/Y, where both w and m are
located in X chromosome, or as Xw m.

h. To show a phenotype, a line is written on top of the symbols of particular alleles. E.g.,
_
B (fly with Bar eye),
_ __
e vg (ebony and vestigial fly)

Description of phenotype does not indicate homozygosity, heterozygosity, and

linkage group.
(B) Maize Genetics

Introduction:
The inheritance of variability in both maize seedlings and aleurone (endosperm)
obeys the Mendelian principles. An expected 3:1 ratio has been observed in a recessive
mutant allele for albinism (absence of chlorophyll) in the maize seedlings and a recessive
allele for colourless aleurone segregating according the Mendel’s first law.
Mendel also postulated that independent assortment of genes occurs when two
traits are considered simultaneously, yielding an expected ratio of 9:3:3:1 among the
dihybrid F2 progenies and dihybrid testcross ratio 1:1:1:1. Indeed, the dihybrid F2 kernels
of corns yield a ratio of 9:3:3:1 for the phenotypes starchy (smooth) endosperm, sugary
(wrinkled) endosperm, colourless starchy endosperm, and colourless sugary endosperm,
respectively.

Objectives:

1. To determines the sexes, indentify and differentiate the morphological traits of the 4
known mutant fly stocks .
2. To recognize and interpret maize F2 data and illustrate Mendel’s law of segregation and
law of independent assortment.

Materials:
Mutant stocks of Drosophila that may be provided are Ebony body colour, Black body
colour, Vestigial wing, Apterous (wingless), Curly wings, Sepia eye colour, Vermilion
eye colour, White eye colour, Apricot (yellow) eye colour, Bar eyes, and Singed bristles,
commercially prepared monohybrid, dihybrid and testcross F2 kernels of corns.
Procedure:

Activity 1 :
The morphological traits of wild and mutant flies were compared. The table 2.1 were completed.
The difference seen between the mutant and wild-type flies with appropriate symbol, sketches, or
description. The wild-types flies that are considered normal was examined for all traits listed
below. A plus sign was placed in the appropriate space to signify the normal condition. The flies
from the given mutant stocks were examined carefully and compared with the wild types. The
mutant flies are wild type for given traits were placed a plus sign in the appropriate space. The
different traits were diagramed.

Activity 2 :
2 possible illustration of endosperm were given in table 2.2 . The number of kernels were
counted without removing from the corn ears in each phenotype. The data were recorded in table
2.3 and then hypothesis were formulated to explain the data collected.
Activity 3 :
The kernels were groups into 4 different types. The number of kernels were counted and
recorded without removing from the corn ears in each of different types of phenotypes in table
2.4

Result :

Experiment 1
Table 1.1 Calculation of x2 for a sample removed from a large population consisting of
equal numbers of coloured and white beads.
Classes Observed Expected Deviations (O − E)2
(O-E)2
(Phenotypes) (O) (E) (O-E) 𝐸
1
Colored 46 44 2 4
11
1
White 42 44 -2 4
11
Total: 88 88 0
X2 = 0.18
Degree of freedom :1
p value : 0.05 Critical value : 3.84
I accept the hypothesis that the data approximate the expected ratio.
Table 1.2 Calculation of x2 on data from tossing coins.
Classes Observed Expected Deviations (O − E)2
(O-E)2
(Phenotypes) (O) (E) (O-E) 𝐸
Heads on
4 7.5 -3.5 12.25 1.6333
both coins
Head on one,
tail on the 14 15 -1 1 0.0666
other coin
Tails on both
12 7.5 4.5 20.25 2.7
coins
Total : 30 30 0

X2 = 4.3999
Degree of freedom : 2
P value : 0.05 Critical value : 5.99
I accept the hypothesis that the data approximate the expected ratio.

Table 1.3 Calculation of x2 for a sample removed from a large population consisting of beads of
two different colors.
Classes Observed Expected Deviations (O − E)2
(O-E)2
(Phenotypes) (O) (E) (O-E) 𝐸
Colored 12 11.25 0.75 0.5625 0.05
White 3 3.75 -0.75 0.5625 0.15
Total : 15 15 0

X2 = 0.2
Degree of freedom : 1
P value : 0.05 Critical value : 3.84
I accept the hypothesis that the data approximate the expected ratio.

Table 1.4 Calculation of x2 on blood group data from a hypothetical population.

Observed Expected Deviations (O − E)2
Phenotypes (O-E)2
(O) (E) (O-E) 𝐸
A 4 10.66 -6.66 44.36 4.16
B 10 2.34 7.66 58.68 25.08
AB 2 0.78 1.22 1.49 1.91
O 10 12.22 -2.22 4.93 0.40
Total : 26 26 0

X2 = 23.23
Degree of freedom : 3
P value: 0.05 Critical value : 7.82
I reject the hypothesis that the data approximate the expected ratio.
Table 1.5 Calculation of x2 for a sample removed from a large population consisting of
beads of two different colors.
Observed Expected Deviations (O − E)2
Phenotypes (O-E)2
(O) (E) (O-E) 𝐸
White 50 46.31 3.69 13.61 0.29
Yellow 127 138.94 -11.94 142.56 1.03
Red 432 416.81 15.19 230.74 0.55
Blue 132 138.94 -6.94 48.16 0.35
Totals : 741 741 23.88

X2 = 2.22
Degree of freedom : 3
P value : 0.05 Critical value : 7.82
I accept the hypothesis that the data approximate the expected ratio.

Experiment 2 :

Table 2.1 Comparison of wild-type and mutant Drosophila.

Trait Wild-type Mutant 1 Mutant 2 Mutant 3 Mutant 4
Body Color Yellow Black line Black line Yellow Black line
Black Yellow Black Yellow Black Black Yellow
Black
Eye Color Red White Red Red Red
Eye Shape
Square shape Square shape Square shape Square shape
with a blunt Round shape with a blunt with a blunt with a blunt
edge edge edge edge
Wing shape Normal Normal Normal Normal Normal
Bristle and Shape Large Small Small Very small small

Table 2.2 Record of F2 data for maize (species 6500) endosperm trait.
Phenotypes Number of Number of Genotypes of
Deviations (O − E)2
kernels kernels the kernels (O-E)2
(O-E) 𝐸
observed expected
Purple 446 443 P_ 3 9 0.02
Yellow 145 148 pp -3 9 0.06
Totals : 591 591 0.08
x2 = 0.08
Degree of freedom : 1
P value : 0.05 Critical value : 3.84
I accept the hypothesis that the data approximate the expected ratio.
The diagram of monohybrid cross and test cross of the species of 6500 maize

Parent : PP x pp

Gametes : P p

F1 : Pp x Pp

Gametes : P,p P,p

F2 : PP,Pp,Pp,pp

Hence, we can determine that the parent genes of PP and pp will produce the F2 offspring with a
ratio of 3:1

Table 2.3 Record of F2 data for maize (species 6502) endosperm trait.
Phenotypes Number of Number of Genotypes of
Deviations (O − E)2
kernels kernels the kernels (O-E)2
(O-E) 𝐸
observed expected
Purple 291 291 P_ 0 0 0
Yellow 291 291 pp 0 0 0
Totals : 582 582 0
2
x =0
Degree of freedom= 1

P value= 0.05 Critical value= 3.84

I accept the hypothesis that the data approximate the expected ratio.

The diagram of monohybrid cross and test cross of the species of 6502 maize

Parent : Pp x pp

Gametes : P,p p

F1 : Pp x pp

Gametes : P,p p

F2 : Pp,pp

Hence, we can determine that the parent genes of Pp and pp will produce the F2 offspring with a
ratio of 1:1
Activity 3:
Let the gene “P” represents the purple traits while gene “p” represents the yellow traits and the
gene P is dominant over the gene p.
Let the gene “S” represents the smooth traits while gene “s” represents the wrinkled traits and the
gene S is dominant over the gene s.

Table 2.5 – Data of independent assortment in maize and calculation of x2 for the maize “6600”
Classes Genotypes Observed Expected Deviations (O − E)2
(O-E)2
(phenotype) (O) (E) (O-E) 𝐸
Purple 339 316.125 22.875 523.266 1.655
smooth P_S_
kernels
Yellow 96 105.375 -9.375 87.89 0.834
smooth ppS_
kernels
Purple 85 105.375 -20.375 415.14 3.940
wrinkled P_ss
kernels
Yellow 42 35.125 6.875 47.266 1.346
wrinkled ppss
kernels
Total: 562 562 0

x2= 7.775

Degree of freedom= 3

P value= 0.05 Critical value= 7.82

I accept the hypothesis that the data approximate the expected ratio.
The diagram of monohybrid cross and test cross of the species of 6600 maize

Parent : PPSS x ppss

Gametes : PS ps

F1 : PpSs x PpSs

Gametes : PS,Ps,pS,ps PS,Ps,pS,ps

F2 : PPSS,PPSs,PpSS,PpSs,PPSs,PPss,PpSs,Ppss,
PpSS,PpSs,ppSS,ppSs,PpSs,Ppss,ppSs,ppss

Hence, we can determine that the parent genes of PPSS and ppss will produce the F2 offspring
with a ratio of 9:3:3:1

Table 2.6 – Data of independent assortment in maize and calculation of x2 for the maize “6602”
Classes Expected Deviations (O − E)2
Genotypes Observed (O) (O-E)2
(phenotype) (E) (O-E) 𝐸
Purple
smooth P_S_ 151 133.5 17.5 306.25 2.294
kernels
Yellow
smooth ppS_ 128 133.5 -5.5 30.25 0.227
kernels
Purple
wrinkled 123 133.5 -10.5 110.25 0.826
P_ss
kernels
Yellow
wrinkled ppss 132 133.5 -1.5 2.25 0.017
kernels
Total: 534 534 0
X2= 3.364

Degree of freedom= 3

P value= 0.05 Critical value= 7.82

I accept the hypothesis that the data approximate the expected ratio.
The diagram of monohybrid cross and test cross of the species of 6600 maize

Parent : PpSs x ppss Written only the offspring

that will produce wanted F2
Gametes : PS,Ps,pS,ps ps

F1 : PpSs x ppss

Gametes : PS,Ps,pS,ps ps

F2 : PpSs,Ppss,ppSs,ppss

Hence, we can determine that the parent genes of PpSs and ppss will produce the F2 offspring
with a ratio of 1:1:1:1

Discussion :

In experiment 1, all x2 analysis are accepted except the activity 4. The rejection of the
hypothesis is due to the small sample of 26 students cannot represents the whole population on
the Earth. A sample must reach a minimum numbers to represents a populations, in this case, we
reject the hypothesis due to this error. In contrary, others x2 analysis can be accepted and obeys
the mendel’s law.

In experiment 2 activity 1, the fly characteristics are determine. We seen a great deviation
of characteristics between the fly species. This is due to the variations of genes in fly. Hence, we
can know that different genes can produce different types of traits in fly. In this experiment, the
comparison of the morphological trait in fly show the variations.

In experiment 2 activity 2, it is the study of the Mendel’s Law of segregation. The law
states that allele pairs segregate during gamete formation, and randomly unite at fertilization.
There are 4 main concepts in this law :

1)A gene can exists in more than one form

2)Organisms inherit two alleles for each traits

3)When gametes are formed through meiosis, allele pairs separate leaving each cell with
a single allele for each traits

4)When the two alleles of a pair are different, one is dominant and the other is recessive.

As an example, for the maize 6500 species, parents cells consist of homozygous dominant gene
(PP) and homozygous recessive gene (pp). The F1 generation will inherit the both of the
dominant gene and recessive gene which resulting a heterozygous genes. When F1 mate with
each other, they will produce a F2 generations with a phenotypes of ratio 3:1 (refer to result in
6500 maize species).

But in the maize species of 6502, when a heterozygous dominant gene parent mates with
a homozygous recessive genes, this mating will produce an offspring of heterozygous gene that
is dominant and homozygous gene which is recessive. Hence, the F2 generations will show a
phenotype with a ratio 1:1 . (refer to results in 6502 maize species)

In experiment 2 activity 3, the experiments are carried out to test the Mendel’s Second
Law which is the law of independent assortment which states that the segregation of alleles of
one allelic pair is independent of the segregation of the alleles of another allelic pair during
gamete formation which involves dihybrid crosses.

As examples is the maize species 6600 and 6602. For species 6600, it is similar to maize
6500 but only introducing one more genes which is known as dihybrid. When a dominant
dihybrid homozygous (PPSS) mates with recessive dihybrid homozygous (ppss), it will produce
a dominant dihybrid heterozygous. When they mates with each other, they will produce f2
generations that obey the Mendel’s Law Second Rule with a ratio of 9:3:3:1.

For the maize species 6602, parents of dominant heterozygous dihybrid genes mates with
homozygous dihybrid recessive gene. The F1 generations will mates with each other that have
different genes (PpSs and ppss)only will produce the F2 generations with the ratio of 1:1:1:1 .

Conclusion :

1. Chi square analysis used to determined whether the data approximating the expected ratio.
2. A Chi square analysis were carried out using a appropriate degree of freedom to compare
with the critical value.
3. The fly were identified by observing their morphological traits.
4. F2 generations of maize can be used to illustrate the Mendel’s First and Second Law.

References :

Bowler, P. J. (1989). The Mendelian Revolution : The EMergence of Hereditarian.

Mendel, G. J. (2009). The Lost and Found Genius of Gregor Mendel, the Father of Modern Genetics. The
Monk in the Garden .