Mendels Approach

Mendel obtained 34 different varieties of peas from local suppliers

Identified 14 strains - 7 specific traits - each with 2 forms
He worked with these strains for 5 years, determining how each character was inherited
The Blending Theory of Inheritance
Mendels experiments tested the blending theory of heredity
Under this theory, a black cat X a white one ?
Mendels Approach Followed the Modern Scientific Method
1. Make initial observations about a phenomenon or process
2. Formulate a testable hypothesis
3. Design a controlled experiment to test the hypothesis
4. Collect data from the experiment
The Scientific Method (continued)
5. Interpret the experimental results, comparing them to those expected under the hypothesis
6. Draw a conclusion and reformulate the hypothesis if necessary
One of Mendels strengths was his careful experimental design
Five Critical Experimental Innovations
There were five features of Mendels experiments critical to success:
o Controlled crosses
o Pure-breeding strains
o Dichotomous traits
o Quantification of results
o Replicate, reciprocal and test crosses
Controlled Crosses Between Plants
Pea plants are capable of self-fertilization and artificial cross-fertilization
Self-fertilization occurs naturally
Cross-fertilization involves removing the anthers from a flower and introducing pollen of the desired type with a
small brush also occurs naturally!
Pure-Breeding Strains to Begin Experimental Crosses
Mendel took 2 years to establish pure-breeding strains
These are strains that consistently produce the same phenotype
Began with crosses between two pure-breeding plants (P generation) that produced offspring called F1 (first filial
generation)
Further Experimental Crosses
Pure-breeding (P) produce F1 of same phenotype
F1 crossed to one another second filial generation (F2)
Crossed to produce the (F3) and so on, as needed
Selection of Single Traits with Dichotomous Phenotypes
Mendel chose dichotomous phenotypes
Easily distinguished - no intermediate phenotypes
Early in his experiments, he worked with an additional trait (#8 seed coat), but noticed a connection with
another trait
Quantification of Results
Mendel counted the number of progeny plants of each type and counted many offspring from each cross
He identified patterns in his results, such as the consistent ratios between phenotypes
The ratios he detected are the foundation of his laws of heredity
Replicate-, Reciprocal- and Test-Cross Analysis
Replicate crosses, produced hundreds or thousands of progeny
Reciprocal crosses, same genotypes were crossed, but the sexes of the parents are reversed
He also performed test crosses
Identifying Dominant and Recessive Traits
All of the traits Mendel studied gave consistent results

Pure-breeding parental strains were artificially cross-fertilized to produce an F 1 generation; self-crossed or intercrossed to produce F2
All F1 produced had the same phenotype, e.g., when yellow-pea and green-pea parents were crossed, all the F 1 had
yellow peas
Dominant and Recessive Traits
The trait shown by the F1 offspring was called the dominant phenotype
The trait not apparent in the F1 was called the recessive phenotype
When F1 were crossed, 75% of the F2 had the dominant trait, the recessive trait reappeared in 25%
Consistent Results of These Experiments
1. Dominance of one phenotype over the other in the F1 generation
2. Re-emergence of the recessive phenotype in the F2 generations
3. A ratio of approximately 3:1 (dominant:recessive) among F2 phenotypes
Evidence of Particulate Inheritance and Rejection of the Blending Theory
Mendels results rejected the blending theory of heredity
Mendel proposed the theory of particulate inheritance
Plants carry two discrete hereditary units for each trait; one of these in the egg and the second in pollen
Alleles
The hereditary particles referred to in the theory are alleles
Together the two alleles for each trait determine the phenotype of the individual
Mendel used letters as symbols to represent the alleles for each trait
Homozygous and Heterozygous Individuals
Pure-breeding individuals have identical copies of the two alleles for a trait
These are called homozygous individuals
The F1 plants had different alleles from each parent and were heterozygous
Consistent Inheritance Patterns among All Traits Studied by Mendel
F1 plants crossed or self-fertilized in a monohybrid cross; (G/g G/g)
3:1 phenotypic ratio is predicted for the F2
1:2:1 genotypic ratio is also predicted ( G/G, G/g, g/g)
Segregation of Alleles
Punnett Square - method of diagramming a genetic cross
The alleles (in gametes) carried by one parent are arranged along the top of the square and those of the other
parent, down the side
Random fusion of the gametes are placed within the square
Mendels First Law
Mendel used his theory of particulate inheritance to formulate the law of segregation (Mendels first law)
His hypothesis describes the units of heredity, their separation into gametes, and the random union of the gametes
into progeny in predictable proportions
The law applies to all seven traits tested
Hypothesis Testing by Test-Cross Analysis
Based on his segregation hypothesis, Mendel expected that half of the gametes of heterozygous F 1 individuals
would carry the dominant allele and half the recessive
He tested this by crossing suspected heterozygous individuals with homozygous recessive individuals from a
pure-breeding stock
He predicted that 50% of the offspring of this test cross should have the dominant trait, and 50% the recessive
Hypothesis Testing by F2 Self-Fertilization
Mendels hypothesis predicts that F2 plants with the dominant phenotype can be homozygous or heterozygous
The heterozygous state (2/3) is twice as likely as the homozygous state (1/3)
He used a self-fertilization experiment to test the predictions of the hypothesis
Results of Self-Fertilization Experiments Agreed with Predictions
Mendel expected that homozygous F2 plants should produce dominant phenotype only
He expected that heterozygous F2 plants would generate a 3:1 ratio of dominant:recessive
These predictions were confirmed by the experimental results

2.3 Dihybrid and Trihybrid Crosses Reveal the Independent Assortment of Alleles

Each of the seven traits Mendel studied showed the same pattern of heredity, explained by the law of segregation
Mendel also studied the inheritance of two or more traits simultaneously

Dihybrid-Cross Analysis of Two Genes

Mendel made dihybrid crosses between organisms that differed for two traits
He began each cross with pure-breeding lines
(e.g., RRGG and rrgg) F1 that were heterozygous (e.g., RrGg).
If assortment is random, four gametes should be equally likely in the F 1 (e.g., RG, Rg, rG, rg)
Mendels Second Law
The 9:3:3:1 ratios in dihybrid crosses illustrate Mendels law of independent assortment
The law states that during gamete formation the segregation of alleles at one locus is independent of the
segregation of alleles at another locus.
Within the 9:3:3:1 ratio, Mendel recognized two 3:1 ratios for each trait
Testing Independent Assortment by Test-Cross Analysis
To test his hypothesis about independent assortment, Mendel performed test-cross analysis
He predicted that the F1 were dihybrid, of genotype RrGr, that crossing them to a genotype rrgg would yield
four phenotypes with equal frequency
Testing Independent Assortment by Trihybrid-Cross Analysis
To test his hypothesis about independent assortment further, Mendel performed trihybrid-cross analysis
The trihybrid cross involved three traits
The cross was: RRGGPP rrggpp; the F1 were RrGgPp
Mendel Compared Predicted Outcomes to Actual Results of the Trihybrid Crosses
The number of gamete genotypes can be expressed as 2 n, where n number of genes
In a trihybrid cross, eight different gametes are possible, each equally likely (1/8)
Using the , expected frequencies for each individual trait, a phenotypic ratio expected for F 2 progeny can be
generated
Trihybrid Cross
3 factor cross examines 3 traits at once:
Punnett square would have 64 squares!
Ratios are 27:9:9:9:3:3:3:1
There must be a better way!

2.4 Probability Theory Predicts Mendelian Ratios

Mendel recognized that chance is the principle underlying the segregation and independent assortment of alleles
of genes at different loci
There are four rules of probability theory that describe and predict the outcome of genetic events
Probability and Genetic Events
Genetic ratios are more appropriately expressed as probabilities
Probabilities range from 0 to 1.0
0 probability - an event has no chance of occurring
1.0 probability - an event is certain to occur
Mathematical definition of probability
P = a/n
where a = the outcome of interest and n = the number of possible outcomes.
P(getting a head on a coin toss)
= 1/2
P(rolling a 1 on a die)
= 1/6
P(drawing the ace of spades)
= 1/52
P(drawing a diamond)
= 13/52 = 1/4
P(having a boy child)

= 1/2
P(dominant offspring from the cross Aa x Aa)
= 3/4
The Product Rule
If two or more events are independent of one another, the likelihood of their simultaneous or consecutive occurrence is the
product of their individual probabilities
This is the product rule, also called the multiplication rule
The product rule (multiplication rule, AND rule)
This rule can be used to calculate the probability of an event when the event is a combination of two or more
events happening together.
P(event A and event B) = P(A) x P(B)
Can only be used for independent events
Two events are independent if the occurrence of one does not affect the probability of the other.
Getting a head on the first flip and a tail on the second flip of a coin are independent.
The product rule (multiplication rule, AND rule)
Drawing a black marble from a bag of black and white marbles (if you put the first marble back before drawing
the second)
If you dont put the first marble back, the two events are not independent.
Examples
P(both heads when flipping 2 coins) = P(head) x P(head)
= 1/2 x 1/2 = 1/4
P(phenotype of round and green in the F2 generation) = P(round) x P(green)
= 3/4 x 1/4 = 3/16
Probability and Genetic Events
Product Rule
(PH:NH) = (1/2)(1/2) = = 0.25
(PH:NT) = (1/2)(1/2) =
(PT:NH) = (1/2)(1/2) =
(PH:NT) = (1/2)(1/2) =
All 4 outcomes occur with equal probability
The Sum Rule
The sum rule is also called the addition rule
It defines the joint probability of occurrence of any two or more equivalent events
The probabilities are summed
Used when more than one outcome will satisfy the conditions of the probability question
The sum rule (addition rule, OR rule)
This rule can be used to calculate the probability of an event when the event of interest can be one of two or more
events.
P(event A or event B) = P(A) + P(B)
Can only be used for mutually exclusive events
Two events are mutually exclusive if the occurrence of one excludes the occurrence of the other.
Getting a head or tail on a coin toss are mutually exclusive
The sum rule (addition rule, OR rule)
Getting a four or a five on a dice roll are mutually exclusive
Getting a four or an even number are not mutually exclusive.
Example of sum rule
P=(1 or 4 on a dice roll) = P(1) + P(4) = 1/6 + 1/6 = 1/3
Forked-line method
Multihybrid crosses
treating the cross as if it were separate monohybrid
apply the product rule to calculate the probabilities
Example using forked-line method
What is the probability of each possible phenotype from the cross YyRr x YyRr ?
An Aid to Prediction of Gamete Frequency
The forked-line diagram is used to determine gamete genotypes and frequencies

Independent Assortment of Alleles from the RrGg RrGg Cross

Mendel predicted that alleles of each locus unite at random to produce the F 2, generating
o round, yellow
R-G()() 9/16
o round, green
R-gg
()() 3/16
o wrinkled, yellow
rrG()() 3/16
o wrinkled, green rrgg
()() 1/16
Conditional Probability
The product and sum rules are used before a cross to predict the likelihood of certain outcomes
Conditional probability involves questions asked after a cross has been made and is applied when information
about the outcome modifies the probability calculation
Conditional Probability
Suppose we cross two Aa parents, where AA and Aa offspring are yellow, while aa offspring are green. Here,
o Pr(AA) = Pr(aa) = 1/4
o Pr(Aa) = 1/2
o Thus, Pr(Yellow) = Pr(AA) + Pr(Aa) = 3/4.
What is the probability that a yellow offspring has genotype Aa?
Conditional Probability
Using our formula,
Pr(genotype = Aa | offspring = Yellow) = Pr(genotype =Aa, offspring = Yellow) / Pr(offspring = Yellow) = (1/2)/
(3/4) = 2/3.
Likewise
Pr(genotype = AA | offspring = Yellow) = (1/4)/(3/4) = 1/3.
The Normal Distribution
In large samples outcomes predicted by chance have a normal (Gaussian) distribution
This is often described as a bell-shaped curve
The Normal Distribution
The mean (m) is the average outcome, and other outcomes are distributed around the mean
The probability of an experimental outcome gets smaller the further it is from the mean
The Probability of Particular Outcomes
Probability of particular outcomes is quantified by a measurement called standard deviation ()
In a normal distribution 68.2% of all outcomes fall within one of the mean, 95.4% within 2, and 99.8% within
3
The Probability of Particular Outcomes
An experimental outcome that is more than 2 from the mean shows a statistically significant difference between
the observed and expected outcome
The Chi-Square Analysis
The chi-square ( 2) test is commonly used for quantifying how closely an experimental observation matches the
expected outcome
o 2 = (O E)2/E;
o 0 = observed values;
o E expected values
Evaluating Genetic Data
Null hypothesis (H0) assumes no real difference between observed and expected ratios
Reject (H0) - deviation from expected is not by chance alone
Fail to Reject or Support (H0) - the deviation may be due to chance alone
Evaluating Genetic Data
Chi-square ( 2) analysis
Used to estimate how often an observed deviation might occur by chance
How well do your observed values fit the expected values
2 = (o-e)2 / e
2 = d2 / e
Evaluating Genetic Data

 Chi-square (2) analysis of monohybrid cross

Coat color in cats (B) black is dominant over (b) gray
Cross heterozygotes (Bb X Bb) = 50 kittens !
Expected progeny (e) Observed (o)
(50) = 37.5
30
(50) = 12.5
20
Evaluating Genetic Data
2 = (o-e)2 / e
Expected (e)
Observed (o)
(50) = 37.5
30
(50) = 12.5
20
(30 37.5)2 / 37.5 = 1.5
(20 12.5)2 /12.5 = 4.5
2 = 6.0
Evaluating Genetic Data
2 = 6.0
So what?
Need to interpret this value
What is the probability that the deviation from expected is by chance?
Evaluating Genetic Data
2 = 6.0
Need to compare to a theoretical value with the same degrees of freedom
Degrees of freedom (df )= n-1
Where n = number of different expected phenotypes
Evaluating Genetic Data
Once you have determined the degrees of freedom, you can assign a p-value (p), or probability
p > 0.05 - the observed deviation occurs by chance alone 5% or more of the time
p < 0.05 - reject the null hypothesis
Evaluating Genetic Data
In our example,
p < 0.05 reject the null hypothesis
The observed deviation is statistically significant not by chance!
Statistical Significance
A statistically significant result from 2 analysis is one for which the P value is less than 0.05
When any experimental result has less than 5% probability, the hypothesis of chance is rejected
P values above 5% indicate a nonsignificant deviation between observed and expected results
This results in failure to reject the hypothesis of chance
2.6 Autosomal Inheritance and Molecular Genetics Parallel the Predictions of Mendels Hereditary Predictions
In the early 1900s biologists began to extend Mendels findings to other organisms
They also began to identify exceptions to the hereditary principles
Mendelian principles can be applied to transmission of certain traits in humans
Human Pedigrees
Family tree representing the pattern of inheritance of a specific phenotype
Four Basic Patterns of Single Gene Inheritance
Pedigrees
Pedigrees, or family trees, are a way of tracing the inheritance of traits in humans
Standard notation is used to indicate males and females, their relationships, and the individuals who show the trait
and those who do not
The generations are indicated by Roman numerals
Autosomal Inheritance

Autosomal inheritance refers to transmission of traits carried on autosomes, chromosomes found in both males
and females
There are two copies of each autosome; so each individual carries two copies of each autosomal gene
Individuals with two identical copies are homozygous; those with two different copies are heterozygous
Autosomal Dominant Inheritance
Only one copy of an allele is necessary for expression.
Heterozygote has a 1 in 2 chance to pass on the allele. Offspring have a 50% chance of being affected.
Homozygous normal offspring have no risk of the disorder or of passing on to their children.
Autosomal Dominant Pedigrees
Transmission is vertical (from generation to generation)
# of affected males and females is equal
Male-to-male transmission is observed
Unaffected individuals have unaffected children
No skipping
Autosomal Dominant Inheritance
1. Each individual who has the disease has at least one affected parent
2. Males and females are affected in equal numbers
3. Either sex can transmit the disease allele
Autosomal Dominant Inheritance, continued
4. In crosses where one parent is affected and the other is not, approximately half the offspring express the disease
5. Two unaffected parents will not have any children with the disease
6. Two affected parents may produce unaffected children
Autosomal Recessive Pedigree
Disorder is rarely present in the parents, collateral relatives or ancestors, but may appear in siblings.
Affects males and females equally.
Consanguinity is more likely to be present in AR pedigrees.
Clinical Characteristics of AR Disorders
Cluster in ethnic groups with geographic or religious isolation and increased consanguinity.
Penetrance is complete - little phenotypic variability.
Most are enzyme abnormalities.
Heterozygotes may be detectable by presence of about 50% of normal enzyme activity
Clinical Characteristics of AR Disorders
Tay Sachs Disease: 1:3,500 Eastern European Jews
Cystic fibrosis (CF): 1:1,600 Caucasians
Sickle cell disease: 1:600 African Americans
Autosomal Recessive Inheritance
1. Individuals who have the disease are often born to parents who do not
2. If only one parent has the disorder the risk that a child will have it depends on the genotype of the other parent
3. If both parents have the disorder, all children will have it
Autosomal Recessive Inheritance, continued
4. The sex ratio of affected offspring is expected to be equal
5. The disease is not usually seen in each generation but if an affected child is produced by unaffected parents, the
risk to subsequent children is
6. If the disease is rare in the population, unaffected parents of affected children are likely to be related to one
another