Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

UNIT 2
APPLICATION OF
MENDEL’S
PRINCIPLES AND
CHROMOSOMAL
BASIS OF
INHERITANCE
Structure
2.1 Introduction Probability and Binomial
Expansion
Expected Learning Outcomes
Laws of Probability
2.2 Chromosomal Basis of Heredity
Binomial Expansion Equation
Sutton and Boveri Hypothesis (1902)
2.4 Formulating and Testing Genetic
Morgan’s Experiment
Hypothesis
Proof of the Chromosomal Theory of
Developing a Hypothesis
Heredity.
Testing Hypothesis
Cellular Basis of Mendelism
2.5 Summary
2.3 Applications of Mendel’s Principles
2.6 Terminal Questions
The Punnett Square Method
2.7 Answers
The Forked Line or Branch Diagram
Method 2.8 Further Readings

2.1 INTRODUCTION
Gregor Mendel postulated particulate nature of heredity and formulated two
fundamental laws of transmission genetics (Refer to unit 1). He published his
work in 1865 which remained unrecognised till 1900. In the intervening period
since his publication, significant advances were made in our understanding of
gamete formation by meiosis and restoration of chromosome number following
fertilisation (gametic fusion). 25
Block 1 Mendelism

In this unit we shall study experimental evidences in support of the

chromosomal basis of heredity, hypothesised by Sutton and Boveri; cellular
basis of Mendelism and applications of Mendel’s principles to predict the
outcome of genetic crosses.

Expected Learning Outcomes

After studying this unit, you should be able to:

 explain Sutton-Boveri hypothesis;

 indicate the major outcome of Morgan’s experiment;

 describe Bridges final proof of the chromosomal theory of heredity;

 explain the chromosomal basis of Mendelism;

 predict the outcome of genetic crosses using different methods;

and

 formulate a null hypothesis and test it using chi square method.

2.2 CHROMOSOMAL BASIS OF HEREDITY

Mendel did not make efforts to popularize his findings except sharing his
results with the famous Swiss botanist Karl Wilhelm Nageli who was studying
cell division and pollen formation. Nageli suggested that both parents make
equal contribution towards the offspring. Although, he had noted that the
nucleus divides during cell division and fertilization brings about fusion of two
parental nuclei but he failed to relate his observations with Mendel’s results.

Several biologists interested in heredity began searching for physical

structures within the cell that could explain the mechanics of gene
transmission. Since gametes are the only cells that pass from parents to the
offspring, biologists began to concentrate on studying the behaviour of the
nucleus and its contents in germinal cells. This was facilitated by significant
advances in light microscopy that enabled biologists to prudently study cell
division. In the 1880s Walther Flemming, a pioneer in cytogenetics, carefully
followed the movement of chromosomes during mitotic cell division and gave
an accurate description of the process. Around the same time meiosis was
discovered in sea urchin eggs by Oscar Hertwig and later described at the
chromosome level in Ascaris (E. van Beneden).

With the rediscovery of Mendel’s work in 1900, Sutton and Boveri

independently proposed the chromosomal localisation of Mendelian factors
based on the behaviour of chromosomes during meiosis. This hypothesis
could explain Mendel’s laws of heredity. The first experimental support for the
hypothesis came from T.H. Morgan who proposed that the gene for white eye
color in Drosophila follows the transmission of the X-chromosome. Later one
of Morgan’s famous students, Calvin Bridges provided the final proof.
26
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

2.2.1 Sutton and Boveri Hypothesis (1902)

Theodor Heinrich Boveri, a German embryologist working with the
roundworm Ascaris, took Flemming's findings to the next level by studying the
behaviour of chromosomes during gametes formation. Ascaris has only two
pairs of chromosomes (2n = 4) that immensely simplified his work and led him
to conclude that the number of chromosomes was reduced to half in the
gametes. He showed that sperms and egg each contributes same number of
chromosomes. The chromosome number is then restored following
fertilization. He also studied dispermic (two sperms) fertilisation of sea urchin
ova which supported the constancy and individuality of chromosomes as such
fusions invariably resulted in abnormal development. At no point did he relate
the behaviour of chromosomes to Mendel’s principles but provided relevant
ideas that were used by Sutton.

Walter Stanborough Sutton, an American graduate student in Edmund B.

Wilson’s lab at Columbia University, studied meiosis in grasshopper
(Brachystola magna) testes. This species has large visible chromosomes of
different sizes. He not only confirmed Boveri’s observation but also described
the configuration of the chromosomes at different stages of meiosis. Sutton T. H. Boveri (1862-1916)
was the first to relate Mendelian genetics with chromosome behaviour. He
suggested that the association of paternal and maternal chromosomes in pairs
and their subsequent separation during reduction division may constitute the
physical basis of Mendel’s laws of heredity (two publications in1902-03).

There are many parallels between genes and chromosomes such as both
occur in pairs; alleles segregate (purity of gametes), so do homologous
chromosomes and different genes undergo independent assortment as they
are located on non homologous chromosomes. But at the outset the scientific
community raised many objections to the hypothesis such as how is one sure
that pairing is between homologous chromosomes or they retain their integrity
through interphase. Above all the hypothesis was not supported by clear cut
experimental evidences. Subsequently it was proved by a combination of W. S. Sutton (1877- 1915)
breeding experiments with supporting cytological data (in some cases).

Now let us answer some simple questions.

SAQ 1
i) What was Walter Sutton’s major contribution?

ii) In which organism did he study meiosis?

2.2.2 Morgan’s Experiment

The chromosomal theory of inheritance appeared to be a reasonable
explanation for how different traits are physically transmitted from parents to
offspring yet there was no experimental evidence supporting it. We shall now
describe experiments carried out by T.H. Morgan with a single spontaneously
arising eye colour mutant in one of his Drosophila cultures; it was a true
breeding white eyed male fly. He initiated crosses between the mutant and red 27
Block 1 Mendelism

eyed flies to determine the inheritance pattern of white eye allele. Like Mendel,
he also performed reciprocal crosses (Fig. 2.1). A glance at his results shows
that reciprocal cross does not yield the same result unlike Mendel’s
experiments with peas. At that time there was no precedence of such a result.

Now let us elaborate the two crosses. In the first cross true breeding red eyed
female fruit flies were mated (crossed) to the white-eyed mutant male fly. He
recorded the results of the cross based on their eye color and sex. All F1
Sib-mating is a progeny flies had red eyes indicating that red eye color is dominant over the
mating of siblings
mutant. When he sib-mated the progeny (inter crossed F1 flies) the ratio of
(brother and sister).
red to white eyes was almost 3:1; the white eyed flies were less than expected
This kind of mating is
due to poor survival. On closer analysis, the white eye phenotype appeared
often used to create
inbred lines.
only in the males. All the females were red eyed and half of the males had red
eyes while the other half had white eyes. He performed test cross of white
eyed males with F1 females and got males and females with both red and
white eyes. This led Morgan to speculate that the inheritance of eye color was
somehow linked to the sex of the fly.

Further confirmation of this hypothesis came from reciprocal crosses. On

crossing white eyed females with red-eyed males he found that all female
progeny flies were red eyed while all male progeny flies had white-eye color.
Inter crossing the F1 flies had the same results as the test cross above.

Original Cross Reciprocal Cross

Fig. 2.1: Morgan’s experiments linked the inheritance of white eyed mutant trait
with the transmission of X chromosome in Drosophila.
28
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

Drosophila has three pairs of autosomes and a pair of sex chromosomes. The
female fly has two X-chromosomes (XX) while the male is XY. Morgan
reasoned that his results can be explained if it is assumed that the gene for
white eye colour is present on the X-chromosome and the Y-chromosome
does not have an allele for it. In addition the dominance of red over white was
clear from his crosses so heterozygous females will mask the recessive trait
which shows up in test crosses or hemizygous males. Based on the
assumptions if we follow the transmission of white eye color it is from father to
daughter (masked) to grandson. Incidentally the experiments with white eye
color mutants also unravelled the characteristics of X-linked recessive pattern
of inheritance. He was the first geneticist to receive the Nobel Prize in
Physiology or Medicine in 1933 for his contributions spanning 17 years since
the discovery of the white eye mutant in 1910.

2.2.3 Proof of the Chromosomal Theory of Heredity

The definitive proof of the chromosomal theory of heredity was provided by
Calvin B. Bridges in 1916. He repeated one of Morgan’s experiments with
white eye colour mutant on a large scale and obtained few exceptional flies.
He crossed white eyed females to red eyed males. As expected majority of
progeny flies were red eyed females and white eyed males but there were few
red eyed males and white eyed females (1in 2000-3000 flies).

The exceptional males were sterile but females were fertile. When he crossed
these females with red eyed males he obtained white eyed daughters and red
eyed sons. Thus the exceptional females could produce many exceptional
progeny flies. These results suggest that exceptional females get both X-
Calvin Blackman Bridges
chromosomes from their mother and males have inherited the X-chromosome
(1889-1938)
from their father. sciencephoto.com

Calvin Blackman Bridges was an American scientist who completed the

proof of the chromosomal theory of heredity. After providing an initial
proof in 1913, he published a detailed account of the relation between
genes and chromosomes in 1916 in a classic paper entitled, ‘Non
disjunction as a proof of the chromosomal theory of heredity.’ This Non disjunction is a
failure of paired
landmark paper was also his PhD thesis.
chromosomes to
Some of his other important contributions in fruit fly genetics include separate (disjoin)
during cell division.
construction of detailed maps of giant polytene chromosomes;
discovered mutants caused by gene duplications and showed that sex
is determined by X/A ratio and not simply by sex chromosomes. He was
also very creative; designed etherizer and replaced rotting bananas with
fly food.

Source: library.cshl.edu/ exhibits/ Bridges.

Calvin Bridges suggested that such a result could happen due to occasional
non- disjunction of X-chromosomes during meiosis in female flies. This would
generate eggs that have either two X-chromosomes or none. When these
abnormal eggs are fertilised with normal sperms it will produce flies with
abnormal sex chromosomes (Table 2.1). 29
Block 1 Mendelism

Table 2.1: Results of Bridges Experiment

Eggs
Sperms Xw Xw O
Xw+ Xw Xw Xw+ Xw+ O
Metafemale (Has Male / sterile
multiple anatomical
abnormalities & poor Red eyes
survival)
Y Xw XwY YO
Female / fertile (Dies)
White eyes
It is clear from table 2.1 that the two types of surviving progeny flies have
abnormal sex chromosome constitution; the females with an additional Y-
chromosome (XXY) and males with no Y (XO). Bridges verified this by
cytological examination thereby proving his assumption. It also supported the
conclusions drawn earlier by Morgan.

2.2.4 Cellular Basis of Mendelism

Now that it is clear that Mendelian factors (genes) are located on
chromosomes, let us relate Mendel’s law of segregation and independent
assortment to the behaviour of chromosomes during meiosis. Fig. 2.2 explains
how the behaviour of homologous chromosomes during meiosis accounts for
segregation of alleles.

Fig. 2.2: Chromosomal basis of the law of segregation.

30
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

Let’s start by considering a plant heterozygous for a pair of alleles (R r) each

of which controls a variant in seed shape / trait, similar to pairs of contrasting
characters studied by Mendel. Now, very carefully follow the movement and
transmission of allelic pair R and r during meiosis. You know that at the time of
gamete formation homologous chromosomes pair up and align at the
metaphase I plate in meiosis. These alleles (R and r) are present on
homologous chromosomes and as they move to opposite poles at anaphase I,
the two alleles R and r are segregated into different gametes (purity of
gametes). The gametes are haploid and inherit only one of the two alleles.
Thus at the cellular level segregation occurs when the pair of
homologous chromosomes carrying alternate alleles separates at
anaphase I.

Now, to understand the independent assortment of genes. Let us consider two

heterozygous gene pairs, Rr and Yy controlling seed shape - round (R/-) or
wrinkled (rr) and seed color - Yellow (Y/-) or green (yy), respectively (Fig. 2.3 ).
The simplest case of independent assortment is when non allelic genes are
present on different (non homologous) chromosomes.

Fig. 2.3: Chromosomal basis of Mendel’s law of independent assortment.

Independent assortment occurs due to random alignment of non homologous

chromosomes at metaphase I, which in turn dictates the pole to which they will
move at anaphase-I. Each homologous pair has two equally likely
arrangements such that either a maternal or paternal homolog may be pulled
to a given pole. In addition, the alignment of non homologous chromosomes is
not dependent on the alignment of any other pair. Implicit in this mechanism is 31
Block 1 Mendelism

a way to generate new combinations; gametes most often have an assortment

of maternal and paternal chromosomes and only very few will get either all
maternal or all paternal chromosomes. Independent assortment alone can
bring about a huge amount of genetic variation even in the absence of
recombination. If n equals the haploid number of chromosomes in a diploid
organism then the possible number of gametic combinations are 2n, for
instance in humans it will be 223 = 8,388,608 possible combinations.

You can visualize random alignment and independent assortment at

metaphase I and anaphase I, respectively in Fig. 2.3. Here we are considering
a cell with two pairs of homologous chromosomes (colored red and blue); one
pair carries alleles (R/r) for seed shape and the other pair has alleles for seed
color (Y/y). There are two possible equally likely alignments for each
homologous pair at metaphase I. You have therefore a total of 4 possible
combinations and gametes with 4 different genotypes are produced in equal
proportion.

Let us check what we have learnt so far.

SAQ 2
i) Assuming the following gene pairs assort independently, predict the
kinds of gametes produced by the following:

a) AaBbccDDEe

b) AABbCcdd

ii) How many different chromosomal combinations will be produced in

Pisum sativum (2n=14) due to independent assortment?

2.3 APPLICATIONS OF MENDEL’S

PRINCIPLES
One of the goals of Mendel and his successors was to predict the outcome of
genetic crosses. In this section we shall describe three general procedures;
the Punnett square method, forked line method and probability method. In
order to use any of these methods to predict the appearance of a trait in
subsequent generations, it is essential to know the genetic basis of a trait.

2.3.1 The Punnett Square Method

An English geneticist Renigald C. Punnett described a shorthand method of
representing all possible outcomes from a genetic cross of known genotypes.
It also allows you to predict the probability with which each genotype and
phenotype can occur in the progeny. This method is generally used when one
or two gene pairs are involved as it becomes increasingly cumbersome to
handle more gene pairs.
32
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

To construct a Punnett square you need to know the genotype of the parents
to predict the possible gametes each parent will produce. It is constructed by
making an M X N grid matrix first, where M represents the number of possible
gametes from the female parent and the number of rows in the Punnett
square, while N represents the number of possible male gametes formed and
forms the columns in the Punnett square.

The next step is to fill the type of gametes produced by each parent. Finally
complete the grid by all possible fusions of male and female gametic cells.
Each cell represents the possible genotype of the progeny/ offspring. An
example of constructing a Punnett square involving two monohybrid parents
(Tt) is depicted in Fig. 2.4. Both parents produce two types of gametes (T and
t) in equal proportion and have equal likelihood to fuse with the gametic cells
of the other parent. The four possible fusions result in three genotypes in 1:2:1
ratio. This is the familiar genotypic ratio of all Mendelian monohybrid selfing. It
is helpful to understand better if you write the phenotype expressed by each
genotype as well. You will note that there are only two phenotypic classes (tall
and dwarf; 3:1) due to complete dominance.

Paternal Gametes

T t

T TT Tt
Maternal gametes

Tall Tall

(1/4) (1/4)

t Tt Tt

Tall Dwarf

(1/4) (1/4)

Fig. 2.4: A Punnett square depicting the outcome of monohybrid selfing (Tt x Tt).
The numbers in the brackets represent the expected proportion of each
genotype in the progeny.

Let us attempt this simple exercise.

SAQ 3
Draw a Punnett square to predict the outcome of:

i) (AaBb X AaBb) crosses

ii) (AaBbCc X AaBbCc) crosses.

33
Block 1 Mendelism

2.3.2 The Forked Line or Branch Diagram Method

The branch diagram is another method of predicting the outcome of crosses
involving two or more genes. Here also as the number of gene pairs increases
it becomes increasingly unmanageable. Let us consider an example of a
trihybrid cross involving three independently assorting gene pairs to show the
proportion of eight phenotypic classes (23) produced from this cross (Fig. 2.5).

Cross: AaBbCc x AaBbCc

The three pairs can be partitioned and each pair segregates in a 3:1 ratio:

Use the forked line / branch diagram method and multiply the individual ratios
to get the proportion of the eight phenotypic classes. It is so named because
of the branching lines.

3C/- 27 dominant (D) for all three

3B/-
1cc 9 D for A & B; recessive (R) for C
3A/-
3C/- 9 D for A and C; R for B
1bb
1cc 3 D for A; R for B and C

3C/- 9 D for B and C; R for A

3B/-
1cc 3 D for B; R for A and C
1aa
3C/- 3 D for C; R for A and B
1bb
1cc 1 R for all three genes
Fig. 2.5: Forked line method.

The final total is 64 (8 X 8 gametic fusions) which merge into eight phenotypic
classes due to complete dominance (Refer to SAQ 3b).

2.3.3 Probability and Binomial Expansion

The probability based method is quicker than the other two methods. It is a
mathematical method of determining the outcomes of a genetic cross. You
know that genetic ratios result from a chance assortment of genes into
gametes and their chance combination into zygotes. Therefore exact
34 predictions are not possible for any event; we only know the likelihood that a
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

particular event will be realised. The two laws of probability help us to

calculate the chances of either two (or more) events occurring together
(simultaneous) or only one at a time (mutually exclusive). Probability is a
fraction between 0 and 1. Let us first consider the laws of probability.

Laws of Probability

The probability theory accounts for the frequency of events. An event is an

outcome of a process. To determine the probability of a particular event, all
possible outcomes of the process must be considered. Then probability of any
event is simply the number of times a particular event happens by the total
possible outcomes. Now let us elaborate the two laws of probability.

The product rule states that the probability of two independent events (A and
B) occurring together is the product of their individual probabilities. It means
that the realisation of one outcome has no influence on the realisation of
others. The word “and” in the statement suggests that you should use the
product rule; for example, human families are equally likely to have either a
boy or a girl at each conception, irrespective of the sex of preceding children.
So the probability of having all three sons, for example, is multiplicative.

P (A and B) = P (A) x P (B)

The sum rule states the probability of occurrence of two independent mutually
exclusive events (Q or R) is the sum of their individual probabilities, provided
there is no overlap in sample space. The word ‘or’ in the statement indicates
that the sum rule should be used.

P (Q or R) = P (Q) + P (R)
Let us refer to a monohybrid cross in Fig. 2.4. You need to remember here
that calculating the probability of gametes of each specific kind in a cross is
similar to calculating the probability of flipping a coin and getting heads or tails.
In this case also there are only two possibilities. Each parental genotype is Tt
and they will form only two types of gametes, T or t in equal proportions. The
gametes from both parents can combine in 4 different ways. Using the product
rule, you can calculate the probability of each progeny. The probability of
homozygous recessive / dominant progeny (tt / TT) is ¼ (½ x ½).

But if you have to calculate the probability of the heterozygous (Tt) progeny,
both product and sum rule is applied. The allele T may come from the egg and
allele t from the pollen and vice versa. Each of these events has ¼ (½ x ½)
chance of occurring. Since both outcomes are mutually exclusive therefore the
combined probability of the heterozygous Tt progeny is ¼ + ¼ = ½. The
probability method of predicting the outcome of genetic crosses is useful when
you are dealing with many gene pairs because the number of possible
combinations increase (Table 2.2).

35
Block 1 Mendelism

Table 2.2: Relationship between pairs of independently assorting alleles,

kinds of gametes; number of genotypes and phenotypes produced in the
F2 generation.

No. of No. of kind of No. of F2 No. of F2

heterozygous gametes (2)n genotypes (3)n phenotypes (2)n
genes pairs, N

1 2 3 2

2 4 9 4

3 8 27 8

4 & so on 16 81 16

Let us do a self check exercise before we begin with Binomial expansion for
determining probability.

SAQ 4
i) In a cross between two individuals of genotype AaBbCc and AaBBCC,
what is the probability that the offspring will be AABbCc or AABBCC?

ii) How many different kinds of F1 gametes, F2 genotypes, and F2

phenotypes would be expected from a cross between AABBCCDDEE X
aabbccddee?

Binomial expansion equation

The binomial expansion is another predictive method for determining the

probability of an unordered combination of events, for instance the
probability that two out of five children (in any order) in a family will be girls.
The binomial expansion equation below is used for repeated trial of events
with constant probabilities:

n!
P  p x q n x
x!(n  x)!

Where,

P = probability that an unordered event will occur

n = total number of events

x =number of events in one category (ex. girls)

p = individual probability of x

q = individual probability of the alternate category (ex. boys)

p + q=1

36 ! = a factorial
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

n! : It is the product of all positive integers down to 1,

for example 5! = 5x4x3x2x1

Note: 0! = 1.

The n! / x! (n- x)! , term in the equation enumerates all possible ways in which
x and (n-x) outcomes can be arranged.

You can also use a multinomial expansion if more than two phenotypes are
involved.

2.4 FORMULATING AND TESTING

GENETIC HYPOTHESIS
Ideally, the first thing to do even before you start an experiment is to form a
hypothesis about the expected outcome(s) of the experiment you have
planned. You may recall Mendel’s hypothesis for independent assortment was
based on results of monohybrid crosses. A hypothesis is a scientifically
testable explanation for a phenomenon based on limited preliminary evidence.
It is an educated guess which is generally the starting point for all further
investigations.

2.4.1 Developing a Hypothesis

We first formulate a null hypothesis (denoted by H0), which is the simplest
probable explanation of your preliminary data. Null hypothesis is generally
believed to be true by default. This should form the basis of your experimental
pursuit. You also formulate an alternate hypothesis (denoted by Ha) i.e. a
hypothesis to justify the results if the experimental approach falsifies the
proposed null hypothesis. Now you should carefully design your experiment
such that the results enable you to differentiate between the null hypothesis
and alternate hypothesis.

Let’s consider a simple Mendelian monohybrid cross to understand this

concept. From our experience with Mendelian crosses, we know that in a
cross between two heterozygous (Aa) plants, the offspring’s are expected to
appear in a 3:1 phenotypic ratio of dominant to recessive trait. In this example
you have used seeds with brown (dominant) and yellow (recessive) colour,
and you want to confirm if brown is dominant over yellow seed colour. This
forms your null hypothesis: Brown seed color is dominant over yellow.

To test your null hypothesis you crossed heterozygous plants and recorded
the seed colour of F2 progeny plants. Out of 400 seeds scored, 295 were
brown and 105 had yellow colour. Now we are ready to test the hypothesis.

2.4.2 Testing Hypothesis

As a researcher, you need to know how much of your experimental result can
differ from the hypothetical figure and still be regarded as statistically close to
expectation. Apply the rules of probability to predict the expected proportion 37
Block 1 Mendelism

and type of progeny. For the above data, your null hypothesis gives expected
numbers of brown to yellow as 300 (400x ¾): 100 (400x ¼).

A simple way to assess how much deviation in the observed numbers is

permissible without casting doubts on the null hypothesis is to use Pearson’s
Chi-square goodness of fit test or simply’ χ2 test ’. This test was introduced
by Karl Pearson to test how well the observed data fits the null hypothesis.

χ2 = ∑ (Observed – Expected) 2 / Expected

Seed colour Brown Yellow

Observed (O) 295 105

Expected (E) 300 100

(O-E) 5 5

Calculate the difference between observed and expected for each phenotype
(O-E). Using the above formula you can test if the observed deviation in the
results is due to chance by comparing the value obtained to a theoretical
distribution.

Let’s calculate the χ2 value:

(a) Brown seeds: (295-300)2 /300 = (5)2 / 300 =0.08
(b) Yellow seeds: (105-100)2/ 100= (5)2/ 100= 0.25

The χ2 for the experiment is the summation of (a) and (b);

0.08 + 0.25 = 0.33

At this point, it is important to choose a critical value i.e. a probability at which

the deviation between the observed and expected value is due to chance
alone. We generally use a critical value of 0.01or 0.05 which means there is a
1% or 5% probability, respectively that the values equal to or greater than the
table value at a given df (degree of freedom, table 2.3) will occur due to
chance.

Another important factor is to determine the degree of freedom (df). The

degree of freedom represents the number of categories or values that can
vary without violating any constraint imposed. For the Chi-square test, the
degree of freedom is equal to n-1, where n = number of expected phenotypic
classes. In the example considered it is (2-1) = 1.

Now let’s go back to the problem. The χ2 value calculated was 0.33. On
comparing the table values at p = 0.01 and one degree of freedom it is 6.64
which is much greater than 0.33. Thus, null hypothesis is not rejected. Smaller
the χ2 value better is the fit. It must be noted that χ2 test does not prove a
hypothesis.
38
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

Table 2.3: Chi square values and probabilities

Degrees P = 0.99 0.95 0.80 0.50 0.20 0.05 0.01

of
Freedom
1. 0.000157 0.00393 0.0642 0.455 1.642 3.841 6.635

2. 0.020 0.103 0.446 1.386 3.219 5.991 9.210

3. 0.115 0.352 1.005 2.366 4.642 7.815 11.345

4. 0.297 0.711 1.649 3.357 5.989 9.488 13.277

5. 0.554 1.145 2.343 4.351 7.289 11.070 15.086

6. 0.872 1.635 3.070 5.348 8.558 12.592 16.812

7. 1.239 2.167 3.822 6.346 9.803 14.067 18.475

8. 1.646 2.733 4.594 7.344 11.030 15.507 20.090

9. 2.088 3.325 5.380 8.343 12.242 16.919 21.666

10. 2.558 3.940 6.179 9.342 13.442 18.307 23.209

15. 5.229 7.261 10.307 14.339 19.311 24.996 30.578

20. 8.260 10.851 14.578 19.337 25.038 31.410 37.566

25. 11.524 14.611 18.940 24.337 30.675 37.652 44.314

30. 14.953 18.493 23.364 29.336 36.250 43.773 50.892

SAQ 5
i) How do you decide the degree of freedom for any given data?

ii) What is the critical value at which we would like to reject the null
hypothesis for three degree of freedom?

2.5 SUMMARY
 After the rediscovery of Mendel’s work in 1900, Sutton and Boveri
proposed the chromosomal localisation of Mendelian factors based on
the behaviour of chromosomes during meiosis.

 The first experimental support to the Sutton-Boveri hypothesis came

from T.H. Morgan who suggested that the gene for white eye color in
Drosophila follows the transmission of the X-chromosome. Later his
student Calvin Bridges provided the final proof to this hypothesis.

 Independent assortment is due to random alignment of non homologous

chromosomes at metaphase I, which in turn dictates the pole to which
they will move at anaphase-I. It also provides a mechanism of creating
enormous variability. 39
Block 1 Mendelism

 Three general procedures have been described to predict the outcome

of genetic crosses; they are Punnett square method, forked line method
and probability method. To be able to predict the appearance of a trait in
subsequent generations it is essential to know the genetic basis of a
trait.

 The probability based method is a quicker method but requires the

correct application of product and sum rules. Many times both rules are
applied to determine the expected outcome.

 The development and testing of a hypothesis using Pearson’s goodness

of fit test (χ2 test) helps us to evaluate how well our data fits the null
hypothesis. Smaller the value of χ2 better is the fit.

2.6 TERMINAL QUESTIONS

1. Describe how the behaviour of chromosomes at meiosis explains the
law of independent assortment.

2. Recall Morgan’s cross of white eyed Drosophila female with red eyed
male discussed in the unit and elucidate. What if in a hypothetical
situation the gene for eye color was located on an autosome? Predict
the phenotype (including the gender) of the F2 flies in this cross.

3. Consider a family of six children and calculate the probability that (a)
exactly four in the family will be girls and (b) at least one is a girl.

4. (a) If you cross two dihybrid parents, what fraction of the family will have
the recessive phenotype for at least one gene?

(b) In a cross between AaBbCC x AABbCc parents, what proportion will

express all three dominant traits?

5. You decide to repeat Mendel’s crosses with peas and crossed two
heterozygous plants with tall and green pod phenotype. The data
generated is given in table below:

Phenotype Number

Tall; green pod 880

Tall; yellow pod 320

Short; green pod 290

Short; Yellow pods 110

a) State the null hypothesis and alternate hypothesis?

b) What is the degree of freedom for this set of data?

c) Calculate the Chi square (χ2) value for this data set.

d) Based on the value will you accept or reject the null hypothesis?
40
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

2.7 ANSWERS
Self-Assessment Questions
1. i) Sutton was the first to relate Mendelian genetics with chromosome
behaviour. He suggested that the association of paternal and
maternal chromosomes in pairs and their subsequent separation
during the reduction division may constitute the physical basis of
Mendel’s laws of heredity.

ii) Grasshopper (Brachystola magna) testes.

2. i) Count the number of heterozygous gene pairs (n) and use the
formula, 2n.

a) n = 3; 23= 8

Gametes are: ABcDE; ABcDe; AbcDE; AbcDe; aBcDE;

aBcDe; abcDE; abcDe

b) n=2; 22 =4

Gametes are: ABCd; ABcd; AbCd and Abcd

ii) The haploid chromosome number of Pisum sativum = 7; 27 = 128.

3. i) Gametes AB Ab aB ab

AB AABB AABb AaBB AaBb

Ab AABb AAbb AaBb Aabb

aB AaBB AaBb aaBB aaBb

ab AaBb Aabb aaBb aabb

ii) Gametes ABC ABc AbC Abc aBC aBc abC abc

ABC

ABc

AbC

Abc

aBC

aBc

abC

Abc

41
Block 1 Mendelism

The type of gametes (23) produced by both parents are filled in the
Punnett square. Now you can complete the 8x8 grid. In part (b) 8 types
of gametes are produced by each parent resulting in 64 gametic fusions.
In both (a) and b) count the number and proportion of different
genotypes.

4. i) Consider one gene pair at a time and calculate the likelihood of

each genotype in the progeny:

Aa x Aa Bb x BB Cc x CC

1AA: 2Aa: 1aa 1Bb: 1BB 1CC: 1Cc

The probability of having the genotype AABbCc is ¼ x ½ x ½

= 1/16

Similarly the probability of AABBCC genotype is ¼ x ½ x ½ = 1/16

The probability of either one of them is 1/16 + 1/16 = 1/8

ii) Kinds of gametes formed by each parent =25

F2 genotypes: (35) = 243

F2 phenotypes: (25) = 32

5. i) Degree of freedom is n -1, where n is the number of phenotypic

classes.

ii) At p = 0.01 the critical value 11.35 will be used and at p = 0.05 the
critical value 7.815 will be used. (Refer table 2.3)

Terminal Questions
1. Refer to subsection 2.2.4 and Fig 2.3.

2. In F1 all progeny flies will be red eyed and in F1X F1 mating they will
segregate in 3:1 red to white eye color, irrespective of sex.

3. a) [(6!) / (4! 2!)].(½)4. (½) 2 = [6x5x4x3x2x1 / 4x3x2x1x2x1] x 1/64 =

5/64

b) The possibilities are that there are 1, 2, 3 or 4 girls. For each

possibility use the binomial expansion (as in part a) and then add
them.

The individual probabilities are 6/64; 15/64; 20/64 and 15/64

The overall probability adds to 50/64

4. a) Of the four F2 phenotypic classes (9:3:3:1), three of them have at

least one recessive phenotype.

A-bb will be ¾ x ¼ = 3/16; aaB- will be ¼ x ¾ = 3/16

aabb will be ¼ x ¼ = 1/16

Each of these is mutually exclusive classes, therefore their combined

probability is 3/16 + 3/ 16 + 1/16 = 7/16

42 b) 1x ¾ x1 = ¾
Unit 2 Application of Mendel’s Principles and Chromosomal Basis of Inheritance

5. a) Null hypothesis: Genes for tall and green trait assort

independently. Alternate hypothesis: Genes for tall and green
trait are linked (present close by on the same chromosome).

b) Three

c) Based on the null hypothesis we expect four phenotypic classes

Tall, green: Tall, yellow: Short, green : Short, yellow in a 9:3:3:1
ratio.

Phenotype Observed Expected (O-E)2 (O-E)2/ E

Tall Green 880 894 196 0.219

Tall Yellow 310 298 144 0.48

Short Green 290 298 64 0.215

Short Yellow 110 99 121 1.22

Total 1590 2.136

Chi square value is 2.136, critical value at p = 0.01 for 3 degree of

freedom is 11.34.

d) We fail to reject the null hypothesis.

2.8 FURTHER READINGS

1. Hartl, D.L and Jones, E.W. Essential Genetics, 4th Ed; Jones and Bartlett
Publishers.

2. Griffith, Antony J.F; Miller, Jeffrey H; Suzuki, David T; Lewontin, Richard

C and Gelbart, William M. An introduction to Genetic analysis, 7th Ed,
2000; W.H Freeman.

3. Snustad, D.P and Simmons, M.J. Principles of Genetics, 3rd Ed, 2003,
John Wiley and sons, Inc.

4. Martins, L.A.-C.P. Did Sutton and Boveri propose the so called Sutton-
Boveri chromosome hypothesis? Genetics and Molecular Biology, 22, 2,
261-71.

43
Block 1 Mendelism

44