University of Science and Technology Houari Boumediene
Faculty of Biological Sciences
L2 - SNV (Natural and Life Sciences)
Module: Genetics
Haploid Genetics
BECHOHRA Louisa, BOULDJENNET Faiza, CHELLOUTI Hiba,
CHEMLLAL Radia, LOUAHCHI Soumia
Introduction
I. Reproductive cycle and growth of Neurospora crassa
II. Examples of allelic differences in Neurospora crassa
III. Genetic analysis of meiosis (case of Neurospora crassa)
III-1. Transmission of a pair of alleles : The Monohybridism
III-2. Transmission of two pairs of alleles : The Dihybridism
II-2.1. Independent genes (The independent segregation)
III-2.2. Linked genes (The genetic linkage)
Introduction:
The haploid cell is a cell that contains only one set of each chromosome (n), in
contrast to the diploid cell, which contains two homologous sets of each chromosome
(2n).
Haploid cell Diploid cell
n=3 2n=6
The Fungi, along with certain lower eukaryotes, are highly useful for genetic analysis
because many of them are haploid.
• Examples of haploid organisms:
This is what Neurospora crassa
looks like when it is molding on
bread.
– Chlamydomonas reinhardtii: unicellular alga
(n=17).
I- Reproductive Cycle and Growth of Neurospora crassa
Neurospora crassa:
A fungus belonging to the Ascomycetes family,
characterized by its filamentous mycelium ;
it has a chromosome number of n = 7;
Its phenotype directly corresponds to its
genotype (haploid type).
The products of meiosis are contained together
within the ascus.
The spores are organized in a specific
arrangement, making it a fungus with ordered
asci.
The fungus Neurospora crassa can reproduce through two modes:
asexual and sexual reproduction.
I- Reproductive cycle and growth of Neurospora crassa
Important Terms
•Perithecium: Sexual reproduction •Mycelium: A network of
organ having flask shaped structures, adjacent cells called hyphae,
containing asci that produce spores. each of which can contain many
nuclei.
• Ascus: An elongated or circular
reproductive cell that contains •Thallus: it is composed of
ascospores. filaments or plates of cells that
forms the mycelium.
Reproductive system Vegetative system
I- Reproductive cycle and growth of Neurospora crassa
1- Asexual reproduction: It is done in the following
manner:
•By dissemination of asexual spores: Reproduction
occurs by the release and spread of asexual spores.
•By fragmentation of the thallus: Reproduction
occurs through the physical fragmentation of the
thallus, which develops into new individuals.
2- Sexual reproduction: This process involves the fusion of sexual spores (conidia located at
the tips of the thalli) that must originate from different thalli with opposing mating types.
Heterothallism: Refers to the presence of distinct types of thalli, specifically thalli of type a
negative (-) and thalli of type a positive (+). This is determined by a pair of alleles, a+ and a-.
Sterile - Fertile
- - crossing crossing
I- Reproductive cycle and growth of Neurospora crassa
Stages of the sexual cycle of Neurospora crassa
1. When two thalli of types a+ and a- come into contact, the haploid cells
combine to form a dikaryon (a dikaryotic cell, which contains two nuclei
within the same cell).
2. The combination of two haploid nuclei (a+ and a) leads fertilization and the
generation of a zygote (cell with 2n chromosomes). The zygote represents
the sole diploid phase of the fungus.
3. The zygote undergoes meiosis, which generates four haploid nuclei.
4. The four nuclei undergo additional mitosis to form eight haploid nuclei,
which constitute the eight spores contained in an ascus, the ascospores.
5. The ascospores released from the ascus germinate and generate new thalli,
which together form the mycelium.
zygote
Dikaryon
(Haplobiontic Cycle)
The products of meiosis are enclosed together in the ascus.
II- Examples of allelic differences in Neurospora crassa
2. Morphological Characteristics
Wild Type Strain: The conidia are black or dark brown, and the thalli are
normal in size.
Mutant Strain: The conidia are pale in color, and the thalli are dwarf-sized.
3. Biochemical Characteristics
The culture medium's chemical composition
Wild Type Strain: It grows on a culture medium consisting of water, sugar,
and vitamins. It is a prototrophic strain.
Mutant Strain: It grows on a culture medium containing water, sugar,
vitamins, amino acids, and nitrogenous bases. It is an auxotrophic strain.
III- Analysis of Meiosis (Case of Neurospora crassa)
The ability of certain fungi, such as Neurospora crassa, to isolate the four meiotic
products in the form of ordered tetrads (the four chromatids) allows for the
determination of the distance between a gene and the centromere of the
chromosome on which it is located.
Furthermore, the isolation of tetrads, whether ordered or unordered, provides
an alternative method for constructing genetic maps and determining the
distances between two or more genes.
III-1. Transmission of a pair of alleles : The Monohybridism
The cross between two strains of opposite mating types (single character = monohybridism)
resulted in offspring consisting of 6 different types of asci.
In each ascus, there are 8 ascospores, with 4 of type a+ and 4 of type a- , arranged in various
patterns. The 8 spores are organized as they were during the tetrad stage in the zygote phase.
Several patterns of ascospore
arrangement within an ascus can occur,
depending on whether crossing-over has
taken place or not.
III-1. Transmission of a pair of alleles : The Monohybridism
Table: Results of crosses between strains of opposite mating types (a+ x a-)
I II III IV V VI
a- a+ a- a+ a- a+
a- a+ a- a+ a- a+ Total
a- a+ a+ a- a+ a- Number
a- a+ a+ a- a+ a- of Asci
a+ a- a- a+ a+ a- =
a+ a- a- a+ a+ a- 240
a+ a- a+ a- a- a+
a+ a- a+ a- a- a+
96 109 9 8 12 6
Neurospora crassa is haploid (n = 7), all cells of the thallus and
the spores are haploid, containing only one allele, either a+ or a- .
The zygote is diploid (2n = 14) and has the a+/a- genotype.
III-1. Transmission of a pair of alleles : The Monohybridism
I II
Interpretation of cross results
a- a+
Cases of the two initial types of asci I and II a- a+
a- a+
There was no mixing of parental characters.
a- a+
There was no crossing-over between the gene and the centromere.
a+ a-
The obtained asci contain 4 a+ spores and 4 a- spores. These asci
a+ a-
are referred to as pre-reduced asci, indicating that the parental alleles
a+ and a were separated during the first meiotic division (Reductional a+ a-
Division, RD). a+ a-
96 109
- -
- -
- - -
- -
- Or - - -
- - -
- - -
-
III-1. Transmission of a pair of alleles : The Monohybridism
Case of the two types of asci III and VI : III IV
a- a+
Parental Character Mixture: the mixing of parental alleles a+ and a- a- a+
results from a crossing-over between chromatids 2 and 3 for ascus III, a+ a-
and between chromatids 1 and 4 for ascus IV (genetic recombination). a+ a-
a- a+
The obtained asci consist of spores arranged in pairs (alternating a+ a- a+
a+ and a- a-). These asci are known as post-reduced asci, indicating that a+ a-
the separation of the alleles a+ and a- occurred during the second meiotic a+ a-
division (Equational Division, ED) as a result of the crossing-over. 9 8
-
-
- -
- - - --
-
- -
- -
- -
- -
- -
III-1. Transmission of a pair of alleles : The Monohybridism
Case of the two types of asci V and IV : V VI
a- a+
Similar to the case of asci III and IV, a- a+
a+ a-
In this instance, crossing-over occurred between chromatids 2 and 4 a+ a-
for ascus V and between chromatids 1 and 3 for ascus VI, resulting in a a+ a-
genetic recombination. The asci obtained in this case are composed of a+ a-
spores arranged in the pattern 2-4-2. a- a+
a- a+
12 6
- -
- -
- - -
1 - 1 - -
2 2
- - -
3
3 - -
4 - 4
-
-
-
-
III-1. Transmission of a pair of alleles : The Monohybridism
Calculate the distance between the gene and the centromere :
Number of post-reduction asci
Post-reduction rate = X 100
Total number of asci (pre- and post-reduced)
Post-reduction rate = ((9 + 8 + 12 +6) / (96 + 109+ 9+8+12+6)) X 100
Post-reduction rate = (35/204) X 100
Post-reduction rate =
a ½ ½
a
a
a
7.29cM
III-2.1. Independent Genes (Independent Segregation)
The cross between two different strains of Neurospora crassa resulted in the following asci.
ASCI
III-2.1. Independent Genes (Independent Segregation)
Interpretation of cross results :
Dihybridism
In this context, the parents have the same genotype as their phenotype, represented as a+b+
(homozygous dominant for both traits) and a−b− (homozygous recessive for both traits).
•Ascus I: This is referred to as a parental ditype because it is composed solely of parental spores.
•Ascus II: This is referred to as a recombinant ditype because it is composed solely of recombinant
spores.
•Asci III and IV: These are called tetratypes because they contain both parental and recombinant
spores.
(
III-2.1. Independent Genes (Independent Segregation)
Stade Octade
Case 1: Ascus I without recombination parental ditype Mitosis a+
Stade Tetrade b+
a+
a+
ED
a+ b+ b+
P1 [a+b+] zygote Prophase I a+
RD b+
b+ a+
a+ a+
a+ a+
a+ b+
a- b+
b+ a- a- a+
b+ b+
b+ b+
a- b-
b-
b- a-
a- b-
b- RD ED
a- a-
a- b-
P2 [a-b-] b- b-
b- a-
a-
b-
b- a-
b-
III-2.1. Independent Genes (Independent Segregation)
Case 2: Ascus II with interchromosomal recombination a+
Mitosis
Recombinant ditype b-
a+
a+
a+
ED
zygote b- b-
P1 [a+b+] Prophase I a+
RD b-
b- a+
a+ a+
a+ a+
a+ b-
a- b-
b+ a- a- a+
b+
b+ b-
b+
a- b-
b- b- a-
a- a- b+
b- RD ED
a- a-
b+
b+
P2 [a-b-] b+ b+
a-
a-
b+
b+ a-
b+
III-2.1. Independent Genes (Independent Segregation)
Case 3: Ascus III - Crossing Over between Gene b and Its Centromere Mitosis a+
Tetratype b+
a+
ED a+
P1 [a+b+] zygote
a+ b+ b+
Prophase I a+
RD b+
a+ b- a+
a+ a+
a+ a+
a+ a+ b-
b+ a- a- b-
a- a- a- a+
b+ b+
b+ b+ b- b-
a-
b- b+
b- b- b- a-
b-
RD ED a- b+
a-
a-
a- b+
P2 [a-b-]
b+ b+
b-
a-
a-
b-
b- a-
b-
III-2.1. Independent Genes (Independent Segregation)
Mitosis
Case 4: Ascus IV - Crossing Over between Gene a and Its Centromere a+
Tetratype b+
a+
a+
ED
P1 [a+b+] zygote
a+ b+ b+
Prophase I a-
RD b+
a+ b+ a-
a+ a+
a+ a-
a+ a- b+
b+ a- a+ b+
a- a- a- a-
b+ b+
b+ b+
b+ b+
a-
b- b-
b- b- b- a+
b-
RD ED a+ b-
a+- a+
b-
P2 [a-b-] a-
b- b-
b- a-
a-
b-
b- a-
b-
III-2.1. Independent Genes (Independent Segregation)
Calculation of distance between genes and their centromeres:
a ½
a
• Post-reduction Rate = (44/200)×100 11cM
• Post-reduction Rate = 22%
a ½
a
b
b ½ 8 cM
• Post-reduction Rate = (32/200)×100
• Post-reduction Rate = 16%
b ½
b
III-2.2. Linked Genes (Genetic Linkage)
The cross between two strains of Neurospora crassa produced the following asci:
ASCI
III-2.2. Linked Genes (Genetic Linkage)
Interpretation of the crossing results:
- Two Traits: Dihybridism
- The genotype of the parents is the same as their phenotype, specifically c+d+ and c−d−
- Ascus I: Parental ditype, consisting solely of parental spores.
- Ascus II: Tetratype, containing a mixture of parental and recombinant spores.
- Ascus III: Recombinant ditype, consisting solely of recombinant spores.
III-2.2. Linked Genes (Genetic Linkage)
Determination of genetic distance (Between Genes c and d): This genetic distance between
genes c and d is proportional to their percentage of recombination.
* Calculation of the percentage of recombination between gene c and d:
The distance between the two genes= d (c–d) = % Recombination = Number of Recombinant Spores X 100
Total Number of Spores
d (c–d) = % Recombinaison = 296 X 100 = 18,5%
200 X 8
So d (gene c – gene d) = 18,5 cM
Linked Genes
Determination of the genetic distance between a gene and the centromere:
Note: The calculation of the distance between the gene and its centromere can only be performed for the post-
reduced gene, which is gene d in this case. Therefore, gene c is not included in this calculation due to the absence
of post-reduced asci.
III-2.2. Linked Genes (Genetic Linkage)
d (gene - centromere) = Post-reduction rate of the gene
2
Total number of post-reduced asci of the gene
Post reduction rate = x 100
Total number of asci (pre- and post-reduced)
Two scenarios:
Genes located on either side of the centromere: d= d1+d2
Genes located on the same side: d=-d2-d1 ou d1= d2-d et d2 =d+d1
III-2.2. Linked Genes (Genetic Linkage)
In our case:
The post-reduced gene is the gene d. Therefore:
Post-reduction rate = (66/200) * 100 = 33
d1 (gene d - centromere) = 33/2 = 16.5 cM
Since d (gene c - gene d) = 18.5 cM:
d = d1 + d2
d2 = d - d1
d2 = 18.5 - 16.5 = 2 cM
The two genes are located on either side of the centromere. Thus, they are separated by the
centromere.
III-2.2. Linked Genes (Genetic Linkage)
Case 1: Ascus I without Crossing Over Parental ditype Mitosis
P1 [c+d+] x P2 [c-d-]
ED
zygote Prophase I
RD
ED
III-2.2. Linked Genes (Genetic Linkage)
Case 2: Ascus II crossing over between chromatids 2 and 3 of gene d Tetratypes
P1 [c+d+] x P2 [c-d-]
III-2.2. Linked Genes (Genetic Linkage)
Case 3: Ascus III double crossing over between chromatids 2 and 3 and chromatids 1 and
4 of gene d Recombinant ditype
P1 [c+d+] x P2 [c-d-]