BIO 4: GENETICS (Laboratory)

Exercise 2
GENE SEGREGATION AND INTERACTION

Gregor Mendel proposed the law of segregation and law of independent assortment by studying the
heredity and variation of seven pairs of inherited characteristics in pea plant (Pisum sativum). Law of
Segregation states that alleles in a gene pair separate cleanly from each other during meiosis. To determine
the principles of segregation, Mendel crossed two plants that differ in one trait. This is illustrated in a
monohybrid cross (one trait being considered, e.g. color) shown below.

P generation Purple x White Phenotype

PP x pp Genotype
P x p Gametes

F1 Generation Pp (Purple Flower)

F2 Generation Purple x Purple
Pp x Pp
Punnet Square Method:
♀\♂ P p
P PP Pp
p Pp pp

Genotypic Ratio: 1PP : 2 Pp : 1pp

Phenotype ratio : 3 Purple : 1 white

When two or more pairs of alleles are considered simultaneously, his second law, the Law of
Independent Assortment, applies. It states that alleles of different gene pairs separate completely and cleanly
from each other and randomly combine during meiosis. He based his second law on the results of his dihybrid
crosses. An example of a dihybrid cross (two or more traits considered) is shown below.

P generation round, yellow seeds x wrinkled, green seeds Phenotype

RRYY x rryy Genotype
RY ry Gametes
F1 generation RrYy (round, yellow)
F2 generation round, yellow x round, yellow
RrYy x RrYy
RY, Ry, rY, ry RY, Ry, rY, ry
Punnet Square Method:
♀\♂ RY Ry rY ry
RY RRYY RRYy RrYY RrYy
Ry RRYy RRyy RrYy Rryy
rY RrYY RrYy rrYY rrYy
ry RrYy Rryy rrYy rryy

Genotypic Ratio: 1 RRYY: 2 RRYy: 1 RRyy: 2 RrYY : 4 RrYy: 2 Rryy: 1 rrYY: 2 rrYy: 1 rryy
Phenotypic Ratio: 9 round, yellow: 3 round ,green: 3 wrinkled, yellow: 1 wrinkled, green
 Exercise 2: Gene Segregation and BIO 4. Genetics
Interaction
A more convenient way of getting the genotypes and phenotypes of the progenies of a given cross
involving several gene pairs is the branching method.

Branching Method: round, yellow x round, yellow Phenotype

RrYy x RrYy Genotype

Finding the gametes: (TEGI – Take each gene pair independently)

Rr x Rr 1 RR: 2 Rr: 1 rr
Punnet Square Method: Yy x Yy 1 YY: 2 Yy: 1 yy

Y Y 1 YY--------1 RRYY
R YR yR 1 RR 2 Yy--------2 RRYy
r Yr yr 1 yy---------1 RRyy
Branching method:
Y---------RY 1 YY---------2 RrYY
R 2 Rr 2 Yy---------4 RrYy
y----------Ry 1 yy---------2 Rryy
Y ------rY
r-------------------- 1 YY--------1 rrYY
y ry 1 rr 2 Yy---------2 rrYy
1 yy---------1 rryy

Genotypic Ratio: 1 RRYY: 2 RRYy: 1 RRyy: 2 RrYY : 4 RrYy: 2 Rryy: 1 rrYY: 2 rrYy: 1 rryy

Branching Method: round, yellow x round, yellow Phenotype

RrYy x RrYy Genotype
(TEPI – Take each phenotype independently)
Rr x Rr 1 RR: 2 Rr: 1 rr 3 round : 1 wrinkled
Yy x Yy 1 YY: 2 Yy: 1 yy 3 yellow : 1 green

3 yellow ------ 9 round, yellow

3 round
1 green ------ 3 round ,green

3 yellow ------ 3 wrinkled, yellow

1 wrinkled
1 green ------ 1 wrinkled, green

Phenotypic Ratio: 9 round, yellow: 3 round ,green: 3 wrinkled, yellow: 1 wrinkled, green

Genes are not merely separate elements producing individual effects, but they could interact with
each other giving entirely different phenotypes. Gene interaction may result to modified phenotypic ratios
deviating from those expected of independently assorting genes exhibiting complete dominance. This could be
allelic interactions involving alleles of a gene pair or non-allelic interactions involving alleles of different
genes.

At the end of the exercise, you should be able to:

 relate chromosomal segregation with Mendelian principles;
 learn to apply the Chi-squared test on observed phenotypic frequencies;
 derive genotypes based on observed phenotypic segregation ratios;
 analyze and determine the type of gene interaction involved in hypothetical crosses; and
 solve problems on probabilities.
 Exercise 2: Gene Segregation and BIO 4. Genetics
Interaction
Suggested Online Resources
 BIO30 Lecture on Gene Segregation and Interaction

 Khan Academy (Introduction to Heredity):

https://www.khanacademy.org/science/high-school-biology/hs-classical-genetics/hs-introduction-to-
heredity/v/introduction-to-heredity

 Khan Academy (The law of segregation):

https://www.khanacademy.org/science/high-school-biology/hs-classical-genetics/hs-introduction-to-
heredity/a/the-law-of-segregation

 Khan Academy (The law of independent assortment):

https://www.khanacademy.org/science/high-school-biology/hs-classical-genetics/hs-introduction-to-
heredity/a/the-law-of-independent-assortment

 Khan Academy (Probabilities in Genetics):

https://www.khanacademy.org/science/high-school-biology/hs-classical-genetics/hs-introduction-to-
heredity/a/probabilities-in-genetics

Laboratory Activity

For activity I, watch a video showing trait segregation in corn using this link:
(https://drive.google.com/file/d/1tNtXRmm-s-AluYaPv0x98dEgRk2MiHOr/view?usp=sharing.) Take note of the
actual number of corn kernels per phenotypic class which will constitute the observed frequency (O). Since,
the expected frequencies (E) are not usually equal to the actual counts (O), we will use the Chi-squared (Χ2)
test to determine the “goodness of fit” between the actual data and the expected results.

Here’s a video showing how to conduct a Chi-squared (Χ 2) test for a sample data
(https://drive.google.com/file/d/1yAEHKy-a8-vJuNm_m6z3Q7fe_jp_8eyj/view?usp=sharing).

Example statement of hypotheses:

Ho: The observed phenotypic ratio fits the expected 9:7 phenotypic ratio for a dihybrid cross that
exhibits independent assortment.

Ha: The observed phenotypic ratio does not fit the expected 9:7 phenotypic ratio for a dihybrid
cross that exhibits independent assortment.

Decision Criteria:
If X2c < X2tab, HO is accepted, otherwise HA is accepted.

I. Gene Segregation in Corn

Ho:

Ha: