Mendelian Principle of Inheritance

2
Dhruti Patwardhan

Genetics is the study of genes and their variation and heredity among organisms.
Long before DNA was recognised as the genetic material, Gregor Mendel through
his studies predicted the presence of such a factor responsible for heredity. Heredity
had been observed in nature for centuries, but Mendel studied this phenomenon in a
scientific manner, performed experiments and put forth his hypothesis that has
withstood the test of time. Although some variations to principles of Mendelian
inheritance have been observed, the basic framework of genetic inheritance initially
proposed by him in essence remains true.

2.1 Mendelian’s Monohybrid Cross

Gregor Mendel is widely regarded as the founder of genetics. He was a priest in an

abbey in Brno where he conducted his experiments on pea plants. Mendel carried out
his experiments on breeding of pea plants for about 7 years from 1856 to 1863 and
presented his findings at the Brno Natural Science meetings in 1865. His paper
which underlined the basic principles of inheritance was published in 1866. How-
ever, Mendel’s work remained unnoticed until 1900 when other scientists like
botanists Hugo de Vries, Erich von Tschermak and Carl Correns obtained similar
results while working independently with plant breeding. They interpreted their
results in the context of Mendel’s theories and published their work supporting
and drawing attention to Mendel’s original work.
Mendel was successful in obtaining and interpreting his results due to a number of
factors. One of the most important factors was his scientific approach and analytical
reasoning. Others before him had crossed plants and described their results. Mendel,
however, was able to formulate a hypothesis based on initial observations and design

D. Patwardhan (*)
Indian Institute of Science, Bangalore, India

# The Author(s), under exclusive license to Springer Nature Singapore Pte 53

Ltd. 2022
D. Kar, S. Sarkar (eds.), Genetics Fundamentals Notes,
https://doi.org/10.1007/978-981-16-7041-1_2
54 D. Patwardhan

suitable experiments to test them. He also recorded the number of plants with
different features across various crosses and tried to interpret these observations to
fit a single framework of inheritance. One of the other reasons for his success was the
choice of garden pea as his experimental model.

2.1.1 The Garden Pea

Pisum sativum, commonly known as the garden pea, was the ideal choice for genetic
breeding experiments. Since Mendel worked in a monastery at Brno, he could easily
access the monastery garden and greenhouse. Pea plants are easy to cultivate and can
grow relatively rapidly with a life cycle of 1 year. He therefore invested years in
following several generations of the plants. Pea plants are also able to produce
numerous seeds which allowed Mendel to calculate mathematical ratios in the traits
of offspring (seeds). Different varieties of peas are available and Mendel was able to
choose those that differed in various traits and were purebred. He also chose to study
features that were present in two easily distinguishable forms/traits like round seeds
versus wrinkled seeds. He avoided those features which had a range of variable
traits. He chose to study seven features which are shown in Fig. 2.1. Apart from
these, some references also mention an eighth feature: seed coat colour which can be
either green or white. Mendel noticed that a coloured seed coat always gives rise to
plants bearing purple flowers, while the white seed coat gave rise to plants having
white flowers. Seed coat colour is therefore sometimes mentioned instead of flower
colour as the traits studied by Mendel.
With advances in molecular biology, studies have been performed to identify the
genes and particular mutations responsible for the traits studied by Mendel. Genes
can be classified into groups based on the structural or functional similarities of
proteins they produce. They may also be grouped together if they all participate in a

Fig. 2.1 The traits or plant characteristics studied by Mendel in his experiments on pea plants:
Mendel studied seven different traits in pea plant for conducting his experiments. Traits being
studied were present in either one of the two forms in the different varieties of pea plant (Pierce
2010)
2 Mendelian Principle of Inheritance 55

Table 2.1 Group symbols and their functions for the traits studied by Mendel: Molecular studies to
understand the traits studied by Mendel have resulted in identification of the genes responsible for
the trait and their function. Of these R, LE, A and I have been cloned and well-studied. Less is
known about the other genes
Trait Group symbol Gene function
Seed shape R Starch branching enzyme 1
Stem length LE GA3 oxidase 1
Flower colour A bHLH transcription factor
Pod colour GP Chloroplast structure in pod wall
Pod form V Sclerenchyma formation in pods
Position of flowers FA Meristem function
Seed colour I Stay-green gene

particular process. Gene group symbol and their functions for the traits studied by
Mendel are given in Table 2.1.
The seed shape can be either round or wrinkled. The seeds differed in their starch,
sugar and lipid content. Wrinkled seeds possess a higher amount of fructose, glucose
and sucrose resulting in higher water retention due to osmotic pressure. A mutation
in the R gene which codes for a starch branching enzyme 1 affected starch biosyn-
thesis. This also further affected the protein and lipid biosynthesis in the seed
ultimately changing its shape. Stem length was controlled by the LE gene. This
gene codes for one of the GA3 oxidase genes which convert the gibberellin to an
active form GA1. Gibberellin is a plant hormone which regulates the development of
the plants including its length. Seed colour is influenced by the I gene which is
important in chlorophyll degradation. Mutation in this gene leads to the appearance
of green seeds. The flower colour is influenced by a gene which exhibits pleiotropic
effects. This means that mutations in the gene can affect multiple traits. The gene
codes for a basic helix-loop-helix (bHLH) transcription factor. It regulates multigene
family-chalcone synthesis (CHS) genes which are responsible for flavonoid produc-
tion. Flavonoids are secondary metabolites produced in plants and are responsible
for pigmentation in plants, thus governing the flower colour. Other genes associated
with the traits studied by Mendel have not been cloned and studied in molecular
detail (Reid and Ross 2011).
To perform his experiments, Mendel crossed different varieties of pea plants. In
order to understand how he achieved this, we need to know a little bit about the plant
reproductive system. The male reproductive organ in a plant is called a stamen. It is
composed of the filament and anther. The filament holds up the anther where pollen
is produced. This pollen is carried by either wind, water or wildlife to the female
reproductive organ. The female reproductive organ is called a pistil which consists of
the stigma, style and ovary. The style holds the sticky stigma at the distal end, while
the ovary is present at its proximal end. Stigma captures the pollen and allows it to
germinate. Sperm carried in the pollen reaches the ovary through tubes formed
during germination. Fertilisation occurs and an embryo is formed which is stored
in the seed capsule. The seed remains dormant until favourable environmental
conditions allow it to develop into a plant.
56 D. Patwardhan

Pea plants often undergo self-pollination. This means that pollen from the flower
will fall on the stigma of the same flower due to its close proximity. This happens
even before the flower has opened. This type of pollination reduces genetic
variability as the pollen and egg come from the same plant allowing them to maintain
their characteristics. Plants which always pass on a specific trait to their offspring are
called purebred varieties. Mendel grew pea plants for around 2 years in this manner
to obtain purebred varieties for each trait. He also wanted to cross plants with
different traits to see what traits were seen in the offspring. To achieve this, he
opened the flowers and removed their anthers to prevent self-pollination. He then
manually dusted pollen from the desired plant on the stigma of a flower from a
different variety. This is called cross-pollination and resultant offspring are called
hybrids. He obtained seeds from these cross-pollinated plants and observed their
traits. He also grew these seeds through the next season to observe the traits of the
hybrid plants (Fig. 2.2).

Fig. 2.2 Figure illustrating the male and female reproductive organs in a plant: To cross different
varieties of plants, Mendel removed anthers from the flower to prevent self-pollination. He then
dusted pollen from desired plant onto the stigma of this flower (Griffiths et al 2011)
2 Mendelian Principle of Inheritance 57

2.1.2 Concept of Dominant and Recessive Traits

Mendel crossed different varieties of plants to study the traits inherited by the
resultant offspring. He started by conducting monohybrid crosses, i.e. crosses
between plants which differed by a single trait. Let us take the example of seed
colour which can be either green or yellow. When Mendel crossed plants having
green seed colour with those having yellow seed colour (referred to as the parental
generation—P), he found that all the offspring called the first filial generation
(F1) had yellow seed colour. He also carried out reciprocal crosses where instead
of taking pollen from yellow seed plant and dusting it on the stigma of a green
seeded plant, he took pollen from a green seeded plant and dusted it on the stigma of
a yellow seed plant. In both cases, he found that plants in F1 generation had yellow
seeds. Similarly, in crosses for the rest of the traits, he found that F1 generation
always showed a single parental trait. It wasn’t a mix of the parental traits, nor did the
outcome change with various repetitions. This trait which was observed in the F1
generation was called the dominant trait and the trait which was lost was called the
recessive trait. Mendel took this experiment one step further and allowed the plants
from F1 generation to undergo self-pollination to create the F2 (second filial)
generation. Most of the plants in the F2 generation had yellow seed colour, but
surprisingly, there were a few plants in which seed colour was green. He counted the
number of these plants and found that the number of plants having yellow seed
colour was roughly thrice the number of plants having green seed colour.
Based on these results, Mendel made certain assumptions and put forth a hypoth-
esis. Although the F1 generation always showed a single parental trait, the second
parental trait reappeared in the F2 generation. This led him to assume that the F1
generation might have received genetic factors for both parental traits. Unless the F1
generation inherited genetic factors from both parents, it is impossible to explain the
appearance of both parental traits in F2 generation. He hypothesised that offspring
must inherit genetic factors from both parents and there must be two genetic factors
in the plant for a single trait. The two genetic factors described here are what we now
know as alleles. Alleles are, simply put, different forms of the same gene and are
designated by a single letter. In this case, the allele for yellow seed colour is
designated as Y and that for green seed colour is designated as y. Since the parental
generation was purebred, the parental generation with yellow seed colour would
have the alleles YY and the one with green seed colour would have the alleles
yy. This composition of the alleles (YY or yy) is referred to as the genotype, and the
trait physically expressed by the plant is called the phenotype (green or yellow seed
colour).
He next assumed that the alleles separate when forming gametes and each allele
gets segregated into one gamete. So the parental yellow seed coloured plants formed
the gametes having allele Y and those from green coloured seed formed the gamete
with allele y. In the F1 generation, these two gametes united and they had the
genotype Yy. All F1 generation only had yellow coloured seeds. This trait was
called the dominant trait. Although the allele for green coloured seed was present, it
was masked and not expressed in the presence of Y. This was called the recessive
58 D. Patwardhan

trait. He concluded that, of the two parental traits, one trait is the dominant and the
other is recessive. Only the dominant trait gets expressed even in the presence of the
recessive trait.

2.1.3 Segregation of Alleles

He further hypothesised that the F1 generation having genotype Yy forms gametes

having Y and y with equal probability. Therefore, half the gametes will have allele Y
and the other half will have allele y and these will get paired randomly in the F2
generation. The resulting progeny might have the genotypes YY, Yy, yY and
yy. Since yellow coloured seed is the dominant character, YY, Yy and yY will
have yellow coloured seeds, and only yy will have green coloured seeds. This
explains the 3:1 ratio of phenotypes observed by Mendel in his experiments. This
ratio can only be obtained if we assume that the alleles get segregated with equal
probability while forming gametes. When the genotype of a plant consists of the
same alleles, they are called homozygous (YY, yy), and when the alleles differ they
are called heterozygous (Yy) (Fig. 2.3).

2.1.4 Mendel’s Analytic Approach

The findings from Mendel’s monohybrid cross are formally stated in two laws
known as law of segregation and law of dominance. Law of segregation states that
during the formation of gametes, two alleles in an individual will separate such that
each gamete will have one allele. Law of dominance states that hybrids of different
alleles will express only one of the parental traits called the dominant trait. Mendel
was able to draw meaningful insights from his work due to the analytic approach
towards his experiments. The ratios Mendel obtained from his experiments were not
perfect. Plants may die or wither before their characteristics can be noted. Some
plants may fail to germinate. Therefore, the ratio of monohybrid cross that Mendel
obtained was almost but not exactly 3:1. However, Mendel obtained numbers for
multiple experiments and noted that the ratios were approximately 3:1 in all cases.
He further went on to self-pollinate plants obtained from F2 generation to confirm
his findings.
Let us take the example here of round vs wrinkled seeds. When plants having
round seeds are crossed with those having wrinkled seeds, the resulting F1 genera-
tion has all round seeds. Here, round seeds is the dominant trait and represented by
the allele R, while wrinkled seeds are represented by the allele r. We can therefore
say that the parental generation had a genotype of RR and rr and the F1 generation
has the genotype Rr. When these F1 plants are self-pollinated, 1/4 seeds are wrinkled
(rr) and 3/4 are round (RR and Rr) in the resulting F2. On further self-pollinating the
F2 generation, he observed that all the wrinkled seeds gave rise to wrinkled seeds on
selfing. This can be explained by the fact that as the genotype of wrinkled seeds is rr,
they will always form gametes carrying the allele r. Therefore, on self-pollination,
2 Mendelian Principle of Inheritance 59

Fig. 2.3 Figure illustrating

monohybrid cross between
plants having yellow and
green seed colour: The
parental generation is
homozygous and produces
only one type of gametes.
These get united in the F1
generation resulting in all
plants having yellow coloured
seeds. The F1 hybrids on
selfing give rise to yellow and
green seeds in a ratio of 3:1.
The genotypic ratio is 1:
2:1 (The Punnett Square
Approach for a Monohybrid
Cross 2020)

they will always give rise to wrinkled seeds. Among the round seeds, 1/3 of the
plants gave rise to only round seeds on self-pollination. The remaining 2/3 gave rise
to a mix of round and wrinkled seeds in the ratio of 3:1. It follows that the seeds
always giving rise to round seeds had the genotype RR. The remaining 2/3 seeds had
the genotype Rr which, similar to the F1 generation, gives rise to the seeds in a ratio
of 3:1. The results of these F3 generations give further evidence to support Mendel’s
hypothesis. These analytic approaches followed by Mendel were one of the strongest
reasons why Mendel was able to come up with a reasonable hypothesis and support
his claims through further experimentation (Fig. 2.4).

2.1.5 Test Cross: One Character

Homozygotes for the dominant allele as well as heterozygotes will display the
dominant trait. To verify if the plant was homozygous or heterozygous for the
dominant allele, Mendel crossed it with a plant showing the recessive trait and
observed their progeny. This cross between plants showing dominant phenotype
60 D. Patwardhan

Fig. 2.4 Monohybrid cross

till F3 generation shows
Mendel’s analytic approach:
Mendel crossed plants with
round seeds and wrinkled
seeds and obtained F1
generation having all round
seeds. On selfing the F1
generation, he obtained the F2
generation in a ratio of
3 round:1 wrinkled as
expected. He further allowed
the F2 to undergo self-
pollination to obtain the F3
generation. He found that
wrinkled seeds always gave
rise to wrinkled seeds. Of the
round seeds, 1/3 always gave
rise to round seeds and the
remaining 2/3 gave rise to
seeds in the ratio of 3 round:1
wrinkled like the F2
generation. This provides
further evidence to support
Mendel’s theory of
segregation and
dominance (Pierce 2010)
2 Mendelian Principle of Inheritance 61

of unknown genotype with a plant of homozygous recessive genotype is called a test

cross. Following Mendelian laws, if the plant being examined is homozygous for the
dominant allele, all its progeny with the homozygous recessive parent will display
the dominant phenotype. In the example below, if the purple flower is homozygous
(having the genotype PP), it will produce gametes having P, and so all the F1 plants
will show purple flowers. If, on the other hand, the purple flower is heterozygous
(having the genotype Pp), it will produce two types of gametes having P or p. These,
on crossing with p from the recessive parent, will give progeny with either purple or
white flowers in a ratio of 1:1.
One of the recessive alleles will be provided by the parent showing the recessive
phenotype as it will only produce gametes with the recessive allele. If a plant appears
in the progeny displaying the recessive phenotype, it is clear that the other parent was
carrying the recessive allele implying that the parent was heterozygous. If all the
progeny show the dominant trait, the other parent has only provided gametes having
the dominant allele implying that the parent was homozygous for the dominant
allele. Test cross is a powerful method to examine unknown genotypes of an
organism showing the dominant trait. This information is useful for breeders who
would wish to choose homozygous plants for further breeding but cannot estimate
the genotype based on observation of phenotype alone (Fig. 2.5).

2.2 Mendelian Dihybrid Cross

We have seen the results of crosses between plants differing in one trait. Mendel’s
next step was to study the pattern of inherited traits in crosses of plants differing in
two traits. Let us take two traits in the pea plant: round vs wrinkled seeds and green
vs yellow seeds. When Mendel crossed green round seeds with yellow wrinkled
seeds, he observed that all the F1 progeny were yellow and round. In the monohybrid
cross for each of the above traits, the F1 progeny expressed the dominant trait which
was yellow colour and round shape. In the dihybrid cross too, the F1 hybrids
expressed the two dominant traits. On selfing the F1 hybrids, he observed four
phenotypes in the F2: yellow and round, green and round, yellow and wrinkled and
green and wrinkled. On counting the number of plants in each category, he surmised
that the plants were approximately in a ratio of 9:3:3:1 for the above combination of
traits.
To make sense of the ratio that he obtained, Mendel made some logical
deductions. He counted the number of yellow vs green and round vs wrinkled
seeds and observed that they were in a ratio of 3:1 similar to the monohybrid
cross. Mendel deduced that of all the F2 plants, 3/4 had yellow seeds and the
remaining 1/4 had green seeds. Of the 3/4 yellow seeded plants, 3/4 had round
seeds and 1/4 had wrinkled seeds. Similarly, 3/4 of the green seeds had round seeds
and 1/4 had wrinkled seeds. This calculation gives the 9:3:3:1 ratio seen above. It
appeared therefore that the dihybrid cross was a combination of 3:1 ratio for two
traits. This will be easier to understand in the branched diagram in Fig. 2.6.
62 D. Patwardhan

Fig. 2.5 Test cross involves crossing of a plant of unknown genotype with a plant showing the
recessive trait: If the parent plant is homozygous, all its progeny will show the dominant trait. If the
parent plant is heterozygous, half of its progeny will show the dominant trait and the other half will
show the recessive trait. In the above illustration, test cross of a homozygous purple flower plant
will result in all its progeny showing purple flowers. Test cross of a heterozygous purple flower
plant will give flowers in the ratio of 1 purple:1 white (Reece et al 2011)

2.2.1 Independent Assortment

Mendel performed the dihybrid cross for a number of combinations of traits and
always got a phenotypic ratio of 9:3:3:1. Let us now understand this ratio in
biological terms. When a plant having yellow wrinkled seeds with the genotype
YYrr is crossed with a plant having green round seeds with the genotype yyRR,
hybrids with yellow round seeds having the genotype YyRr are produced. These
hybrids can produce four gametes having four different combinations: YR, Yr, yR
and yr. Each gamete carries one allele for each trait. On self-pollination, these
gametes can merge in a variety of different combinations giving a phenotypic ratio
of 9:3:3:1 as seen in the figure below (Fig. 2.7).
The fact that the dihybrid cross ratio is a combination of 3:1 ratio for each trait
tells us that the gametes for each trait can assort independently. It means that allele Y
has equal probability of pairing with either allele R or r to form a gamete. If one of
the alleles in the cross above assorted preferentially with another allele, we would
2 Mendelian Principle of Inheritance 63

¾ round seeds ¾ X 3/4 = 9/16 yellow round

seeds
¾ yellow seeds
¼ wrinkled seeds ¾ X ¼ = 3/16 yellow
wrinkled seeds

¾ round seeds ¼ X ¾ = 3/16 green round

seeds
¼ green seeds
¼ wrinkled seeds ¼ X 1/4 = 1/16 green
wrinkled seeds

Fig. 2.6 Phenotypic ratios obtained in a dihybrid cross: Each of the traits gives a 3:1 ratio. In the
example above, we obtain seeds in a ratio of 3 yellow:1 green seeds. Each of these phenotypes also
shows a ratio of 3 round:1 wrinkled seeds. This gives the overall 9:3:3:1 phenotypic ratio seen in a
dihybrid cross

not obtain a phenotypic ratio of 9:3:3:1. This is called the law of independent
assortment which states that different gene pairs can assort independently during
gamete formation. However, genes which are close to each other on the same
chromosome do not assort independently because they are held together on the
same chromosome. In this case, alleles for different genes which are on the same
chromosome always assort together during meiosis. The modified law of indepen-
dent assortment can therefore be stated as ‘Gene pairs present on different
chromosomes assort independently of each other during formation of gametes’.
The tendency of genes which are close to each other to be inherited together is
called linkage. Genes which get inherited together are classified into a single linkage
group. Therefore, if any of the genes studied by Mendel belonged to same linkage
group, their phenotypic ratios would have differed from the ones defined by Mendel.
Mendel did not observe linkage between the genes that he studied and hence put
forth the law of independent assortment. Mendel’s work has been criticised on the
basis that his data fits too well with his hypothesis and does not show as much
variation due to chance as expected. Mendel’s critics also cite the lack of evidence of
linkage as one of the reasons to doubt Mendel’s work. Recent work has, in fact,
shown that the seven traits that Mendel studied belong to five different linkage
groups of which only stem length and pod form show strong linkage. Mendel might
have been lucky in his choice of traits for dihybrid cross. He might not have
performed dihybrid crosses for this particular combination of stem length and pod
form, or he would have been surprised by the resulting phenotypic ratios. Seed shape
and pod colour show weak linkage, and all other traits are not linked allowing
Mendel to obtain the same ratio for most of his dihybrid crosses. The debate about
the validity of Mendel’s work is discussed in detail in Box 2.1.
64 D. Patwardhan

Fig. 2.7 Genotypes of progeny obtained from a dihybrid cross for wrinkled yellow seeds with round
green seeds: The F1 progeny has yellow round seeds and can produce four types of gametes. These
are shown on the top row and first left column of the square on the right. The various combinations of
these gametes give a progeny with a phenotypic ratio of 9:3:3:1 (Griffiths et al. 2011)
2 Mendelian Principle of Inheritance 65

2.2.2 Test Cross: Two Characters

Similar to the monohybrid cross, Mendel performed test cross for the dihybrid cross
to verify his conclusions. As stated above, test cross involves crossing a plant of
unknown genotype with a plant homozygous recessive for the traits under consider-
ation. If we were to perform a test cross for the F1 produced from the dihybrid cross
above, the tester (homozygous recessive individual) would be a plant having green
and wrinkled seeds. In this case, we would expect the F1 to form gametes with all the
combinations, RY, Ry, rY and ry, according to the law of independent assortment.
This when fertilised with ry gametes from the tester would give the following
different combinations: RrYy (round, yellow), Rryy (round, green), rrYy (wrinkled,
yellow) and rryy (wrinkled, green). A phenotypic ratio of 1:1:1:1 would be expected.
This is the result that Mendel obtained from his test cross for two characters
providing evidence for his law of independent assortment.
The F1 hybrid obtained above is heterozygous for both traits. Let us see how the
results would differ if the plant being tested was homozygous for either of the traits.
If we take a plant having yellow, round seeds, its genotype could be either YyRr
(discussed above), YYRr, YyRR or YYRR. If it is YYRR, it will produce only one
type of gamete YR which when crossed with yr will produce all plants of YyRr
genotype and single phenotype of yellow round seeds. If it is YYRr, two types of
gametes will be produced: YR and Yr. This will give two phenotypes on test cross:
yellow, round seeds (YyRr) and yellow, wrinkled seeds (Yyrr) in a ratio of 1:1.
Similarly, if the genotype is YyRR, it will produce two phenotypes on performing
test cross: yellow, round seeds (YyRr) and green, round seeds (yyRr) in a ratio of 1:
1. In this manner, we can detect the genotype of the individual based on the number
and ratio of phenotypes produced (Fig. 2.8).

2.3 Mendelian Trihybrid Cross

Mendel’s laws can be extended to obtain genotypic and phenotypic ratio of a

trihybrid cross, i.e. a cross between plants differing in three traits. As the number
of traits increase, the complexity of the ratio increases and several different methods
can be used to predict the results. Let us take the example of three genes A, B and C
having recessive alleles a, b and c. If a homozygous dominant plant with genotype
AABBCC is crossed with a homozygous recessive plant aabbcc, the F1 hybrid will
have a genotype of AaBbCc. When the F1 plant is further self-pollinated, it will
produce eight different gametes as shown below (Fig. 2.9).

2.4 Application of Mendelian Principles

Mendel’s principles of segregation, dominance and independent assortment can be

applied to analyse inheritance of multiple traits. Various methods like Punnett
square, forked line method or probability method can be utilised to obtain
66 D. Patwardhan

YYRr YyRr YyRR

YYRr yyrr YyRr yyrr YyRR yyrr

yr yr yr

YR YyRr YR YyRr YR YyRr

Yr Yyrr Yr Yyrr yR yyRr

yR yyRr

yr yyrr

Fig. 2.8 Test cross for a dihybrid cross can reveal the parental genotype: Yellow round seeds with
three different genotypes (YYRr, YyRr and YyRR) give rise to different phenotypes and pheno-
typic ratios in the progeny when subjected to a test cross. This allows us to estimate the genotype of
the unknown individual. (Adapted from Klug et al. 2012)

Fig. 2.9 Gametes produced during a trihybrid cross: Eight different gamete combinations of alleles
and therefore eight different types of gametes may be produced by an individual heterozygous for
three gene pairs (Klug et al. 2012)
2 Mendelian Principle of Inheritance 67

phenotypic and genotypic ratios of cross between multiple traits. We will obtain the
genotypic ratios for the trihybrid cross mentioned above using both Punnett square
and forked line method.

2.4.1 The Punnett Square Method

In the Punnett square method, a grid is created with the gametes from one parent on
the upper side and those from the other parent on the left side. Each cell or block
within the grid contains a combination of alleles from both parents giving the
genotype of the offspring resulting from the combination of the respective gametes.
It is named after Reginald C Punnett who devised this method. It is a tabular
representation of all possible combinations between the maternal and paternal
alleles.
In the image below, gametes produced from F1 hybrid of a trihybrid cross are
represented on the top row and left column of the grid. Since this is a self-pollination,
the gametes on both sides are identical. In case the parents differ in genotype,
gametes from one parent will be in the top row and will differ from gametes
produced by the other parent which will be on the left column. The first box in the
grid shows the genotype AABBCC which is produced if the gametes ABC and ABC
(corresponding parental gametes in first row and first column) were to combine. In
this manner, all possible genotypes of the progeny can be represented in the Punnett
square in a simplified manner. The genotypes showing the same phenotypes are
represented by the same colour of the cell in the grid. From this, we can infer that the
phenotypic ratio of a trihybrid cross will be 27:9:9:9:3:3:3:1 (Fig. 2.10).

Fig. 2.10 Punnett square showing genotypes produced from a trihybrid cross: The genotypic and
phenotypic ratio for a cross between individuals of genotype AaBbCc and AaBbCc is shown in the
Punnett square above. The gametes produced from each parent are shown in the top row and left
column. Each grid represents a combination of the gametes in the respective row and column. The
individuals showing the same phenotype are in the same colour. We can see that a phenotypic ratio
of 27:9:9:9:3:3:3:1 is obtained
68 D. Patwardhan

2.4.2 The Forked Line Method

Although useful, Punnett square can be too cumbersome to use when more than
three traits are being analysed. In such instances, forked line method, also called the
branch diagram, can be very useful. In this method, the genotypic or phenotypic
outcome for one gene pair is first predicted. Then, outcome of the next gene pair is
computed in conjunction with earlier gene pair. This method is followed for all the
remaining gene pairs. In the figure below, phenotypic ratio of trihybrid cross is
predicted using the forked line method. Here, three traits are considered: round vs
wrinkled seeds, green vs yellow seeds and grey-brown vs white seed coat. We know
the dominant phenotype for the other traits except seed coat where grey-brown is
dominant over white. According to Mendel’s law of segregation and random
fertilisation, a monohybrid cross between round and wrinkled seeds will result in
3/4 seeds being round and the remaining 1/4 being wrinkled. This is the outcome for
the first trait. Now, of the round seeds, 3/4 seeds would be yellow and 1/4 green,
which is the outcome for the second trait. Similarly, 3/4 of the wrinkled seeds will be
yellow and 1/4 green. Next, of the round and yellow seeds, 3/4 will have grey-brown
coat and 1/4 will have a white coat. This gives us a total of 27 round, yellow and
grey-white coat seeds (3/4 round
3/4 yellow
3/4 grey-brown coat ¼ 27/64).
Similarly, proportion of round, yellow and white seed coat will be 3/4
3/4

1/4 ¼ 9/16. We can calculate the proportion for all phenotypes in this manner and
obtain a phenotypic ratio of 27:9:9:9:3:3:3:1.
Calculations for genotypic ratio can be done in a similar manner. Although the
phenotypic ratio for a monohybrid cross is 3:1, we need to bear in mind that the
genotypic ratio is 1:2:1 (1 AA, 2 Aa and 1 aa). The genotypic calculation for a
trihybrid cross using forked line method is illustrated below. There are a few rules of
thumb that can be used to cross-check your calculations. First, count the number of
heterozygous gene pairs in the cross. In a cross AaBbCc X AaBbCc, heterozygous
gene pairs 3. 2n will be the number of different gametes that can be produced from
each parent. 3n will be the number of different genotypes produced after fertilisation,
and 2n will be the number of different phenotypes produced in this cross. In the
above example, therefore, 23 ¼ 8 different gametes are formed from each parent,
33 ¼ 27 different genotypes are produced in this cross and it gives rise to 23 ¼ 8
different phenotypes. These are the numbers that we get from our calculations with
the forked line method as well as Punnett square method (Fig. 2.11).

2.4.3 The Probability Method

Probability is a mathematical tool that predicts the likelihood of occurrence of an

event.
We can easily use probability to predict the outcome of a genetic cross. Probabil-
ity can be calculated as the number of times an event occurs divided by the total
number of events. For example, the probability of picking a queen of hearts from a
deck of cards would be 1/52. This is because there is only one queen of hearts (one
2 Mendelian Principle of Inheritance 69

Fig. 2.11 Forked line method for obtaining genotypic and phenotypic ratios in a trihybrid cross:
For a monohybrid cross, genotypic ratios of 1:2:1 are obtained. Then for each of the genotypes, the
ratio for the next set of genes is calculated and further on and the proportions multiplied at the end.
Similarly, phenotypic ratios can be calculated by multiplying 3:1 ratios for each trait (Klug et al.
2012)

event) in a deck of 52 playing cards (all possible events). If, however, we were to
calculate the probability that a card picked would be any queen, this probability
would be 4/52 as there are four queen cards in a deck. There are two rules to be
followed for the calculation of slightly more complicated probabilities.

1. Multiplication rule: The probabilities of co-occurrence of two or more events

can be calculated by the product of their individual probabilities. This rule can
only be used if the events are occurring independent of each other. If the
occurrence of one event affects the probability of occurrence of the next event,
then their combined occurrence cannot be a simple product of their independent
probabilities. For example, the probability that two consecutive rolls of a dice are
a three and a six in that order is 1/6
1/6 ¼ 1/36. The individual probabilities of a
three and a six in a roll of dice are 1/6 and 1/6, respectively. Their product will
give the probabilities of their co-occurrence. We have already used the multipli-
cation rule while calculating the dihybrid and trihybrid cross ratio in the branch
diagram or forked line method. Multiplication rule is applied when the word and
is used. In the above example, we wanted to find the probability of ‘a three and a
six in a roll of dice’.
2. Addition rule: We can calculate the probability of either of two or more mutually
exclusive events occurring together by the addition rule. For example, if we just
wanted to obtain the probability of occurrence of a three or a six in a roll of the
dice, we would find the sum of individual probabilities. Thus, the probability is
1/6 + 1/6 ¼ 2/6 ¼ 1/3.
70 D. Patwardhan

The multiplication and addition rules can be used in predicting the outcome of
genetic crosses instead of Punnett square or forked line method. Let us consider
the cross between two plants having round seeds with genotypes Rr and Rr. The
probability of wrinkled seeds can be calculated using multiplication rule. The
probability of obtaining r allele from one parent is 1/2 and from the other parent is
also 1/2. For a wrinkled seed, the genotype needs to be rr and its corresponding
probability is 1/2
1/2 ¼ 1/4. If we were to calculate probability of round seeds,
both multiplication and addition rules need to be used. Round seeds can occur
because of three genotypes: RR, Rr and rR. Their individual probabilities are as
follows:

1=2 R
1=2 R ¼ 1=4 RR

1=2 R
1=2 r ¼ 1=4 Rr

1=2 r
1=2 R ¼ 1=4 rR

Their combined probabilities would therefore be 1/4 + 1/4 + 1/4 ¼ 3/4 round
seeds. It is easier to use probability method for calculation of complex crosses with
multiple traits as compared to Punnett square or forked line method.

2.4.3.1 Binomial Theorem

Binomial theory can be utilised when we want to calculate the occurrence of a
specific set of outcomes among a large number of potential events. Let us consider
the case of galactosemia. In this disorder, mutation in one of the galactose
metabolising genes prevents the individual from converting galactose to glucose.
The affected individual may show symptoms like lethargy, failure to gain weight and
liver damage. This disorder can only occur if both copies of the gene are mutated.
Thus, the parents of a child may each be carrying one mutated allele and not express
the disorder. Only if the child inherits both mutated copies will he/she be affected.
Let us assume that the gene affected here is a and its wild-type/normal allele is A. If
we now want to find the probability that both kids of heterozygous parents are
affected. The probability of one child being affected (aa) is 1/4 (Aa X Aa ¼ 1/4 aa,
3/4 A-) and that of two children being affected will be 1/4
1/4 ¼ 1/16.
Now, if we suppose the couple has three children and we want to find the
probability that two children are unaffected and one child is affected. There are
three scenarios in which this is possible:

1=4
3=4
3=4 ¼ 9=64 ðchild 1 affected, other 2 unaffectedÞ

3=4
1=4
3=4 ¼ 9=64 ðchild 1 and 3 unaffected, child 2 affectedÞ

3=4
3=4
1=4 ¼ 9=64 ðchild 1 and 2 unaffected, child 3 affectedÞ

Total probability ¼ 9=64 þ 9=64 þ 9=64 ¼ 27=64

2 Mendelian Principle of Inheritance 71

This calculation becomes more complex for situations with more number of
children and multiple different combinations. If we want to find the probability of
this couple having five children, three of whom are affected and the remaining two
are not, we can use the binomial expression. The binomial expression is of the form
(a + b)n where a and b are probabilities of two alternate events and n is the number of
times the event occurs. In the above case, we can define a as the probability that the
child suffers from galactosemia (1/4), while b is the probability that the child remains
unaffected (3/4). n here is the number of children which will be 5. The binomial can
be expanded as follows:

ða þ bÞ5 ¼ a5 þ 5a4 b þ 10a3 b2 þ 10a2 b3 þ 5ab4 þ b5

It follows the rule:

ða þ bÞn ¼ an þ an1 b þ an2 b2 þ an3 b3 þ . . . :bn

The expansion of (a + b)n consists of n + 1 terms. Each of these terms has a

numerical coefficient. The coefficient of the first term is always 1. The second term
has the coefficient same as the power to which the binomial is raised. So, in this case,
it is 5. For the next coefficient, multiply the coefficient of the previous term with the
exponent of a in that term. Divide this by the number of the term in the equation.
Thus, to calculate the coefficient for a3b2 in the above expansion, (5
4)/2 ¼ 10
where coefficient of earlier term is 5 and exponent of a in that term is 4 and it is the
second term in that equation. Similarly for the coefficient of a2b3, it can be calculated
as (10
3)/3 ¼ 10. We can calculate the coefficients for the rest of the terms and
expand the binomial.
Another method to calculate the coefficients in the equation is to use Pascal’s
triangle. We can determine the coefficients for each term in the binomial expression
from the terms in front of the corresponding n. Notice that all terms other than 1 in
Pascal’s triangle are the sum of terms directly above them (Fig. 2.12).
Once we have the equation, we can obtain the probability of any combination of
events by simply inserting the values of a and b. For example, to obtain the
probability of three out of five children having galactosemia, the term we use is
10a3b2:

10a3 b2 ¼ 10
ð1=4Þ3
ð3=4Þ2 ¼ 90=1024 ¼ 0:087

In this manner, we can easily calculate the probability of any combination of

events. There is another method to do the above calculation. It uses the formula:

n!
P¼
as bt
s!t!
P is the overall probability of co-occurrence of two events X and Y. Event X has a
probability of a occurring s times, while event Y has probability b occurring t times.
72 D. Patwardhan

Fig. 2.12 Pascal’s triangle: Pascal’s triangle can be used to obtain coefficients for terms in the
binomial expansion for any n. The terms other than 1 in Pascal’s triangle are a sum of the terms
directly above them

In the above case, X is the probability that the child is affected. Therefore, a is 1/4
and s is 3. Event Y is the probability that the child is unaffected. Here, b is 3/4 and t is
2. N is the total number of events which is 5 in this case. The symbol ! is for a
factorial which is the product of all positive integers from 1 to n. For example, 5! ¼ 5

4
3
2
1.
The calculation therefore is:
5!
P¼
ð ¼Þ 3 ð ¾Þ 2
3!2!
5
4
3
2
1
¼
ð¼Þ3 ð¾Þ2
ð2
1Þð3
2
1Þ
¼ 0:087

This value is the same as that obtained from binomial theorem.

2.5 Test of Genetic Hypothesis

Crosses between two individuals of known genotypes yield a certain genotypic and
phenotypic ratio. Based on Mendel’s laws, we can predict a certain ratio. However,
the experimental ratios may not match the expected values. Other than technical
difficulties (like death of plants before the phenotype can be observed), chance plays
a very important role in this deviation. This is easily illustrated with the example of a
coin flip. We know that the probability of getting a heads or tails in a coin flip is 1:1.
2 Mendelian Principle of Inheritance 73

If we do the coin toss for a large number of times, say 1000, we can expect that we
will get a number close to 1:1. However, if we toss the coin only ten times, we might
get seven heads and three tails or two heads and eight tails. This deviation from
expected ratio is just a chance event.
Genetic ratios however can also be different, if there is some linkage between the
traits being studied or if the gene is following some non-Mendelian pattern of
inheritance. An experimenter needs to know if the deviation from expected ratios
is just a matter of chance or it is of some biological significance. In such cases, we
can make use of a chi-square test.

2.5.1 The Chi-Square Test

Chi-square test, also written as χ 2 test, is used to evaluate how well the observations
support the null hypothesis. It is calculated from the sum of squared errors or sample
variance. A chi-square test can only tell us if the resulting ratio of genetic crosses is
deviating from the expected ratio merely due to chance. It cannot tell us if there is a
mistake during crossing or during calculation of expected ratios or there are some
complex inheritance patterns involved. In other words, it gives us a probability that
the difference in observed and expected ratio can be due to chance alone.
Let us take an example to understand how to use the chi-square test. A monohy-
brid cross between two tall plants resulted in a progeny of 100 tall plants and 40 short
plants. If we were to assume that the genes involved followed a Mendelian inheri-
tance pattern, we would expect a ratio of 3 tall:1 dwarf plants. For a total of
140 plants, 3/4
140 ¼ 105 plants should be tall and 1/4
140 ¼ 35 plants should
be short. We see that the observed ratio differs slightly from the expected ratio. Is
this merely an effect of chance?
We start by establishing a null hypothesis (Ho). The null hypothesis is called so
because it assumes that there is no real difference between our expected and
observed outcomes and any deviation is a result of chance events. Through the
probability derived from the chi-square test, we can then accept or reject the null
hypothesis. Our null hypothesis for this example will be that the inheritance follows
a ratio of 3:1. The formula for chi-square test is:

ðO  E Þ2
χ2 ¼ Σ
E
where E ¼ expected value for that category
O ¼ observed value for that category
Σ ¼ sum of calculated values for all categories
Plugging in the above values in this equation:
74 D. Patwardhan

ð100  105Þ2 ð40  35Þ2

χ2 ¼ þ
105 35
¼ 25=105 þ 25=35
¼ 0:238 þ 0:714 ¼ 0:952

After this calculation, we have to determine the degrees of freedom (df) which is
n  1 where n is the number of different categories the value may fall into. Here, the
plant can be tall or short. Thus, n ¼ 2 and df ¼ 1. Degrees of freedom are considered
because more categories introduce more deviation in the results. We now have to
interpret the χ 2 value in terms of its corresponding probability value. This calculation
is very complex, and we instead make use of a standard table which provides
probability values for different χ 2 values for each degree of freedom.
In the table below (Fig. 2.13), we can see that the calculated value of 0.952 for df
1 lies between p value of 0.5 and 0.1. We can interpret this as the probability that the
observed deviation from expected value is due to chance is between 10 and 50%.
Traditionally, scientists have accepted a p value cut-off of 0.05. That is to say that if
the p value is above 0.05, we can accept the null hypothesis. If the p value is less than
0.05, it means that the probability that the deviation is due to chance is less than 5%.
In this case, the null hypothesis is rejected. In our example, we can accept the null
hypothesis and conclude that the variation seen in the ratios is a product of chance
and that the inheritance indeed follows a 3:1 ratio.

Fig. 2.13 Probability values for χ 2 distribution: Figure giving probability values for estimated
χ 2 values at different degrees of freedom. The probability value keeps decreasing towards the right,
while the χ 2 values keep increasing
2 Mendelian Principle of Inheritance 75

Let us take another example. In a cross between plants having violet flowers and
white flowers, violet flowers were observed in F1. On self-fertilisation, it was seen
that 790 of the progeny had violet flowers and 210 had white flowers. Can we
ascertain if this follows the Mendelian pattern of inheritance?
Null hypothesis—The pattern of inheritance follows Mendelian genetics and does
not differ from a ratio of 3:1:

ðO  E Þ2
χ2 ¼ Σ
E
E: If the flower colour inheritance followed Mendelian genetics, we would see
that 3/4 of the total flowers would be violet and 1/4 would be white since violet is the
dominant character (because F1 flowers were violet). The expected numbers would
therefore be:
Violet 3/4
1000 ¼ 750, white 1/4
1000 ¼ 250

ð790  750Þ2 ð210  250Þ2

χ2 ¼ þ
750 250
¼ 2:133 þ 6:4 ¼ 8:533

Degree of freedom here too is 1, as only two characters are being observed.
P value for χ 2 8.533 at df 1 is less than 0.005
We can therefore reject the null hypothesis. The probability that deviation in ratio
is purely due to chance is very less and the gene is probably following some
non-Mendelian pattern of inheritance.

2.6 Application of Mendelian Principles in Human Genetics

Mendel’s work has shed light on the inheritance of genes and traits. We can use this
knowledge to analyse inheritance of various genetic diseases and traits in humans
too. We can do this by obtaining information regarding occurrence of the trait being
studied in the family of the affected individual.

2.6.1 Pedigree Analysis

Pedigree analysis is similar to a family tree for a specific trait. It is basically a chart
which illustrates which family members have the traits being studied. This aids in
understanding the method of inheritance of the trait. We can also predict the possible
genotype of individuals for that trait which can help in predicting probability of
inheritance of the trait in future generations.
A set of standardised symbols are used for illustrating a pedigree. Squares
represent males, and circles represent females. A shaded box denotes individuals
that expressed the phenotype being studied. A horizontal line between two
76 D. Patwardhan

individuals denotes mating. Their progeny are represented in the order of birth on a
horizontal line connected to the parental mating line. Different generations are
represented on descending levels. A double line connecting two individuals denotes
consanguineous marriage. A marriage between second cousins or even more closely
related individuals is referred to as a consanguineous marriage. Many studies have
shown that consanguinity is one of the major contributors of birth defects and
abnormalities. If an individual has a recessive gene, his progeny might inherit the
gene but not express the phenotype. Thus, individuals belonging to the same family
have a greater probability of carrying the recessive gene. A marriage between
members of a family increases the probability of a child from this union inheriting
two copies of a recessive gene and therefore suffering from a genetic disorder
inherited in an autosomal recessive manner. Although consanguineous marriages
have reduced over the years, they are still prevalent in the Middle East and parts of
Asia and Africa. More symbols and their meanings are given in Fig. 2.14.
Let us examine the pedigree shown in Fig. 2.15. Individual 4 from generation III
is the proband. This means that this individual was the first to be investigated for this

Fig. 2.14 Standard symbols

used while drawing a pedigree
chart: Proband denotes the
first individual studied in the
family for this trait.
Consanguineous marriage
means marriage between
individuals from the same
family, usually first
cousins (Klug et al. 2012)
2 Mendelian Principle of Inheritance 77

Fig. 2.15 Pedigree showing autosomal recessive mode of inheritance: The pedigree shown above
shows that the character has skipped a generation. Fewer members of the family are affected and the
trait is evenly distributed between males and females. Based on this, we can conclude that the mode
of inheritance of the gene being studied is autosomal recessive (Klug et al. 2012)

phenotype and prompted construction of the pedigree. We can see that one of the
siblings of individual 4 is also affected. None of the parental generation (Gen II) has
any affected members. Among the grandparental generation (Gen I), individual 1 is
affected. We can draw a few conclusions from this information. First is that the trait
being studied is recessive. Based on Mendel’s law of dominance, if the trait was
dominant, at least one parent of the affected individual would have expressed the
trait. Since none of the parents show the trait, they are most likely carriers of the
recessive allele. This skipping of generation in expression of traits is a characteristic
feature of recessive traits. Second, although there aren’t many affected individuals,
the trait seems to be passed equally between males and females (Gen III individuals).
We can therefore assume that the recessive trait is on an autosome. Of the 23 pairs of
chromosomes that are present in humans, 22 are autosomes and 1 pair is a sex
chromosome (X and Y chromosomes). This means that the 22 pairs are inherited
randomly between males and females. However, the sex chromosomes determine
the sex of the individual. In humans, XX determines a female and XY determines
males. Therefore, the inheritance of traits present on sex chromosomes will not
follow Mendelian patterns and instead show different ratios for males and females.
For example, genes on Y chromosomes will only be passed on to males and not to
the females. The probability of inheritance of a mutated gene between males and
females remains the same for an autosomal disorder irrespective of which parent
carries the mutated gene. In case of recessive disorders, however, the probability of
inheritance of mutated gene between the sons and daughters will differ based on
whether the father or the mother is carrying the mutated gene. We will discuss this
further in the next chapter. For the context of this discussion, it is enough to
78 D. Patwardhan

understand that any trait which seems to be passed equally between males and
females is most likely present on an autosome.

2.6.2 Mendelian Segregation

We can also deduce from this pedigree that either individual I-3 or I-4 was hetero-
zygous for the allele being studied. For individual III-4 to be affected, both his
parents need to be heterozygous for the allele in question. Based on Mendel’s law of
segregation, for individual III-4 to be homozygous recessive, he has to inherit one
recessive allele from each of his parents. Individual II-3 could have obtained the
recessive trait from individual I-1 since he was affected. For individual II-4 to be a
carrier, either individual I-3 or I-4 would have to be a carrier as they do not show the
phenotype. We can determine the pattern of inheritance and composition of the
genotype of an individual from a pedigree based on Mendel’s laws of dominance and
segregation.
Some examples of autosomal recessive disorders are cystic fibrosis, sickle cell
anaemia and Tay-Sachs disease. Cystic fibrosis is caused by a defect in both copies
of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The bodily
fluids become thick and sticky. Due to this, the individuals suffer from respiratory
and digestive problems. The abnormal mucous clogs airways and damages the
pancreas. Tay-Sachs disease is a progressive neuronal disorder that affects the
neurons in the brain and spinal cord. It is a rare disease in which infants start
showing symptoms after 3–6 months. Their development slows and they develop
muscle weakness. Progression of disease leads to loss of hearing, paralysis and
seizures. The disease is caused due to two defective copies of the HEXA gene. This
gene codes for the hexosaminidase A enzyme that plays a role in the breakdown of a
fatty substance called GM2 ganglioside. Build of GM2 ganglioside is toxic for the
neurons.
Let us now take the example of an autosomal dominant disorder. A typical
pedigree is shown in Fig. 2.16 for inheritance of an autosomal dominant trait. We
can immediately observe that this pedigree has at least one affected member in each
generation. This is a typical characteristic of inheritance of a dominant allele. We can
also see that the disorder has been passed on to both the males and the females. We
can therefore infer that the allele is present on the autosomes. An example of
autosomal dominant disorder is the Marfan syndrome. Marfan syndrome affects
the connective tissue due to which a number of abnormalities in the heart, bones,
joints, eyes and blood vessels can be observed. Marfan syndrome patients are tall
and slender with long narrow faces. Their arm span exceeds their body height and
they have elongated fingers and toes. Marfan syndrome is caused by a mutation in
the FBN1 gene which codes for the fibrillin 1 protein. Fibrillin 1 is instrumental in
the formation of microfibrils. Microfibrils are threadlike filaments that provide
strength and flexibility to the connective tissue. They also bind to growth factors
2 Mendelian Principle of Inheritance 79

Fig. 2.16 Pedigree showing autosomal dominant mode of inheritance: The pedigree shown above
has an affected member in each generation. A number of members of the family are affected and the
trait is evenly distributed between males and females. Due to this, we can assume that the mode of
inheritance of the trait being studied is autosomal dominant (Griffiths et al. 2011)

and control their release. Absence of functional fibrillin 1 reduces the amount of
microfibrils leading to lack of control in the availability of growth factors. An
excessive amount of available growth factors leads to overgrowth and abnormal
tissue formation. Being an autosomal dominant disorder, the presence of even one
mutated allele is sufficient for the manifestation of this disease.
Neurofibromatosis type 1 is also an autosomal dominant disorder. It is associated
with a range of symptoms. Individuals suffering from the disease show a pigmenta-
tion change with appearance of dark patches of the skin. Benign tumours
(non-cancerous) grow along the nerves in the brain and other parts of the body. In
some cases, these tumours may turn cancerous. Additionally, these individuals may
suffer from hypertension, macrocephaly, skeletal abnormalities and abnormal cur-
vature of the spine. Some affected individuals may develop learning disabilities or
attention deficit/hyperactivity disorder (ADHD). Neurofibromatosis type 1 is caused
due to mutations in the NF1 gene which codes for a protein called neurofibromin.
This protein is produced in the neurons as well as glial cells like oligodendrocytes
and Schwann cells. Neurofibromin acts as a brake for cell division and is known as a
tumour suppressing gene. Non-functional neurofibromin leads to lifting of this brake
and rampant and uncontrolled cell division leading to formation of tumours as seen
in the disease.
80 D. Patwardhan

Observations of the pedigree charts given above will make it clear that we do not
always see expected Mendelian ratios in these inheritances. This is mainly because
we do not have a large number of progeny which can be observed to reach the
expected ratio. The inheritance of gametes is dependent on chance, and as discussed
in chi-square analysis, we can see vastly different ratios than expected for a small
sample size. The second factor is that in a population, some alleles are more
commonly found than others. Most people are carriers of the rare allele and very
few are homozygous for the rare allele. Thus, mating usually happens between
individuals who are either heterozygous or homozygous for the most common allele,
making the appearance of individuals homozygous for the rarer allele very
uncommon.

2.6.3 Genetic Counselling

Pedigree analysis can also be used to predict the probability of the progeny inheriting
a certain trait or disease. Couples having certain disorders running in the family or
who themselves are affected may wish to know the probability of their children
inheriting the disease. Couples with one of their children affected with a genetic
disorder may seek to understand the possibility of their next child having the disease.
Genetic counselling may help in such situations. Genetic counsellors will obtain
information from the couples about affected family members and draw a pedigree.
From this, they can deduce the mode of inheritance and further calculate the
possibility of their unborn offspring inheriting the disorder. They can provide
information and educate as well as address concerns of the family members regard-
ing the disorders and provide support. They can also inform individuals about their
genetic predisposition to certain diseases and lifestyle changes if any that can prevent
or manage the disorder.
The Human Genome Project completed in 2003 was a 13-year-long study aimed
at sequencing the entire human genome. This sequencing was carried out at multiple
labs around the world and DNA was taken from a number of donors. The sequence is
therefore a mosaic and not from any one individual. This prompted the 100,000
genome project in the UK which aims at sequencing 100,000 individuals comprising
people with rare diseases, their families and cancer patients. With the mapping of
these genomes, we can hope to understand more and more about our genes and the
functions that they play in health and disease. We may be able to pinpoint the causes
of a number of genetic diseases which remain unknown till now. Genomic sequences
from patients will aid in developing diagnostics and therapeutics for individuals
suffering from Mendelian disorders. It may allow us to get closer to personalised
medicine where the analysis of an individual’s genome may provide clues as to what
treatment would be most effective for the individual.
2 Mendelian Principle of Inheritance 81

Box 2.1 Scientific Concept: Gregor Mendel’s Genetic Experiments:

A Statistical Analysis After 150 Years (Jan Kalina)
Gregor Mendel is not only regarded as the founder of genetics but also as one
of the pioneers of applying statistical principles in their experiments. Mendel’s
parents were peasants and bred fruit trees. After completing his secondary
education, Mendel joined the St Thomas Augustinian abbey where he was
ordained as a priest after some years. The abbey fostered an interest in science
and encouraged Mendel to carry out his plant breeding experiments for which
he was also given the use of one of the greenhouses in the abbey. In 1866,
Mendel published his landmark paper which defined basic laws of genetics.
He could not, however, enjoy the recognition of his work in his lifetime.
Mendel passed away in 1884, aged 61 years, due to chronic nephritis. His
work was rediscovered in 1900 independently by three scientists. In 1936,
Robert Fisher reanalysed Mendel’s data in light of the new statistical methods
available. After a detailed analysis of the data, he concluded that Mendel’s
data seemed to be too much in agreement with his theoretical expectations.
Given the variation caused by chance, his data seemed ‘too good to be true’.
He therefore concluded that Mendel must have falsified most, if not all, of his
data to fit his theoretical assumptions (Kalina 2016).
Since then, a large number of papers have come out analysing both sides of
the debate. In 2008, many scientists from different fields came together to
write a review on the subject hoping to put an end to the argument. The authors
were able to refute all arguments except the too good to be true claim for the
data. There were few Mendel supporters like Pilgrim, who found fault with the
way in which Fisher had performed his analysis (Pilgrim 1986). However, his
arguments were later refuted by Edward (Edwards 1986). Then, there was a
group of people who believed Fisher’s analysis but found it too stringent and
sought ways to analyse the data in an alternative manner. They too however
found that Mendel might have adjusted the data to suit his hypothesis. Then
there are those who believe that Fisher’s analysis holds true given the
assumptions. However, they try to provide suggestions for explaining the
high p values other than deliberate malpractice. Some of the explanations
provided are stopping experiments when the results seem to be good, error
carried out by some anonymous assistant and discarding plants due to suspi-
cion of some errors like pollen contamination and data selection for presenta-
tion (Novitski 2004).
In spite of the range of explanations provided, the bias towards expected
values seen in Mendel’s data has not been completely resolved. New statistical
models to explain the bias are being proposed to provide a satisfactory
explanation and end the controversy regarding falsification in Mendel’s data.
Based on the distribution of p values observed in Mendel’s data and different
simulation approaches, a plausible explanation of a two-stage model has been

(continued)
82 D. Patwardhan

Box 2.1 (continued)

put forth. It states that Mendel repeated some of his experiments, presumably
those which showed the largest deviation from expected values. He then
reported only the best of two or a combination of results of the two which
would result in values closer to those that were expected. This is called as a
two-stage model where experiments were performed and results evaluated in
second stage again for those experiments which did not meet the distribution.
This speaks of an unconscious bias that Mendel introduced in his experimental
approach but cannot be called as intentional scientific fraud (Pires and Branco
2010; Kalina 2016). Mendel’s laws have stood the test of time and are still
accepted today. His data might have been biased or adjusted but his
conclusions about the nature of genes and their inheritance are still relevant
today. We must therefore focus on Mendel’s contribution to the field of
genetics and quantitative biology and put an end to the debate as we seem to
have some possible explanations.

2.7 Summary

• Gregor Mendel’s painstaking decade long experiments and theories derived from
them have laid the foundation of genetics, and he is known as the father of
genetics. Mendel’s three laws of genetics provide a framework for understanding
the inheritance of genes. His work was rediscovered independently by three
botanists in 1900.
• In monohybrid crosses, plants which differed in only one trait were crossed. The
F1 hybrids carry the alleles for both parental traits but only express one of the
parental traits which was termed as the dominant trait. The parental trait that was
not expressed in the F1 hybrid was termed as the recessive trait. This was called as
the law of dominance.
• The F1 hybrid produces gametes possessing the dominant and recessive alleles
with equal probability. These can get paired randomly in F2 generation. This was
called the law of segregation of alleles.
• Mendel also carried out dihybrid crosses where he crossed plants differing in two
traits. He observed that the traits were inherited independently of each other. This
was referred to as the law of independent assortment.
• He developed the method of test cross to determine the genotype of plants. Plants
showing a dominant trait may either be heterozygous or homozygous for the
dominant allele. Test cross involved crossing the plant in question with a plant
showing the recessive trait. The ratio and phenotype of progeny from this cross
could indicate heterozygosity or homozygosity of the plant being studied.
• Phenotypic and genotypic ratios for a cross with multiple traits can be predicted
by using the methods of Punnett square, forked line method or probability
2 Mendelian Principle of Inheritance 83

method. Statistical methods like chi-square goodness of fit test can be used to

determine if the observed ratio from a cross vary from the expected ratios purely
due to chance.
• Application of Mendelian principles can contribute to understanding and
predicting genetic disorders in humans. Pedigrees can be constructed for a
particular trait to understand mode of inheritance of a gene. Genetic counselling
may help in counselling parents suffering from genetic disorders who want to
understand the chances of their children inheriting the disease.

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