6 Genetic
1.5 Tetrad analysis which spend the great.test
and fungi,
Certain species of
life cycle in the
lower ryotes,
particularty
been used
unicellularalgae
in mapping
studies. The sac
reproduction.
fungi
(ascomycetes parthaveof beenthe
have also of sexual
haploid state, style
of their unique
particularly useful to geneticists because
Haptoid cell
Haploid cel
(ID) Diplold zygote
Zygotic meiosis
Tetrad of cells contained
within an ascus
Mitosis
Figure 1.20
DO Sexual reproduction in ascomycetes.
For simplicity, this diagram shows
each haploid cell as having only one
chromosome per haploid set. However,
fungal species actually contain several
chromosomes per haploid set.
Fungi may be unicellular or multicellular. Fungal cells are
typically haploid (1n) and can reproduce asexually. In
addition, fungi can also reproduce sexually by the fusion of
two haploid cells to
diploid zygote can then proceed through melosis to create a diploid zygote (2n). The
of four spores is known as a tetrad. In
produce four haploid cells, which
are called spores. This group
some species,
meiosis is followed by a
cells, known as an octad. The cells of a tetrad or mitotic division to produce egnt
octad are contained
within a sac known as an
ascus (plural, ascl)
Ordered or unordered tetrad and octad
The arrangement of spores within an ascus varies from
Space for the tetrads or octads of species to species. In
spores to
These occur in fungal species such as S.
randomly mix together. This is some cases, the ascus provides en gh
cerevisiae. By known as an unordered tetrad or octad.
ehat orevents spores from randomiy comparison, other species o
moving around. This can create
of fungi produce a very tight
ascus
a linear tetrad or octad found in N. sa.
o
GenetiCS 37
(3)
Saccharomyces cerevisiae Neurospora crassa
Flgure 1.21 Dierent arrangements of fungal spores.
A key feature or i n e a r t e t r a d s or octads is t h a t the position and order of spores within the ascus r e n e e
lationship to each other as they were produced by meiosis and mitosis. This idea is schematically shown in ngu
1.22. After the original diploid cell has undergone chromosome replication, the first meiotic division produces
cells that are arranged next to each other within the sac. The second meiotic division then produces four ces
are also arrangea in a straight row. Due to the tight enclosure of the sac around the cells, each pair of daugnte
oy
cells is forced to lie next to each other in a linear fashion. Likewise, when each
sis, each of the daughter cells is located next to each other.
of these four cells divides
Mitosis
Meiosis II
Meiosis
spores. After fertilization,
the zygote's
1.22 Formation of a linear octad in N. crassa. Neurospora has ordered
Figure is forced to lie along the long
axis
ascus is narrow, the two nuclei
nucleus undergoes meiosis I. Since the Neurospora axis of
also oriented along the long
the second meiotic division, which is
of the ascus. The two nuclei then undergo
a r e ordered according to
their centromeres.
the ascus. The result is that the spores
tetrad
1.5.1 Analysis of ordered
centromere. This approach has
the distance between a gene and the
Linear tetrad analysis can be used to map
crassa. In N. crassa, the products
of meiosis are contained in an ordered array
been extensively exploited in N.
each pair representing one of the products
contains eight ascospores in four pairs,
of spores. Each mature ascus
each gene with respect to its
of meiotic product makes it possible to map
of meiosis. The ordered arrangement
and its centromere. Two c a s e s are
the recombination frequency between a gene
centromere; i.e. to determine
there is a crossover between the locus and its
centromere.
on whether or not
possible depending
First case
between a gene and its centromere, the alleles of the gene (for example
A and a)
In the absence of crossing over
Octad contains
meiotic division, this separation is called First Division Segregation (FDS).
must separate in the first
cells that contain
a linear of four haploid cells carrying the A allele, which are adjacent to four haploid
arrangement
an allele i.e. 4:4 arrangement
of spores within the ascus (figure 1.23).
38
Genetics
A
4
AL A
Mitosis
a
a
Segregation of homologous
chromosome during meiosis I 4
Segregation of sister
chromatids during meiosis II
Figure 1.23 First Division Segregation (FDS): No crossing over produces a 4:4
arrangement.
Second case
If a cross
over does occur between the gene and its
second meiotic division. centromere, the A and a allele do not
This separation is called Second Division become separated
deviates from the 4:4 Segregation (SDS). So, second division unti
pattern. Depending on the relative
crossover, the ascus will contain a 2: locations of the two
chromatids
segrega
is a result of
crossing over, the 2:2:2or 2: 4:2 pattern (figure 1.24 and that
participated in
centromere and the gene of percentage of SDS asci can be
used to calculate
1.25). Since a
pattern of 5
interest. As shown in the the map distance between
centromere if it begins in the figure, a
crossover will
2:2:2:2 or 2: 4:2 patternregion between the centromere and that gene.only separate a gene from its orig
depend on the distance
between the gene of Therefore, the chances of gettin
interest and the centromere.
C
entromere
of interest and
t h a l
crossover,
the ascus W the
percentaoe the
figui
in c e n t r o m e r e
and
over, s h o w n
gene
of crossing As the
S a result i n t e r e s t .
the between
the gene oof b e t w e e n
centromereand
d i s t a n c e
region
the
centromere if it begins depend
on
2 pattern
2 : 2 : 2or 2:
4
Mitosis
Crossing over in Segregation of
Segregation of sister
prophase I of h o m o l o g o u s c h r o m o s o m e
chromatids during
meiosis I during meiosis I
meiosis II
produces a 2: 2: 2: 2 arranaem.
A single crossing
over ment.
igure 1.24 Second Division Segregation
(SDS):
AL
A
Mitosis
4
Crossing over in
prophase I of A
meiosis I Segregation of
homologous chromosome
during meiosis I Segregation of sister
chromatids during
Figure 1.25 Second Division meiosis II
Segregation (SDS): Asingle
crossing over
produces a 2 4: 2 arrangem
Genetics 39
Calculation of map distance
The percentage of asci with SDS patterns
for a gene can be used to map the genes with respect to its
To deternmine the map distance between the centromee
centromere and a gene, we first count the number ot sDs
the
tota number of asci. In
SDS asci, only half of the
as a
actually
the of a crossover.
product
the map distance is calculated as:
spores
are
Ihereto
xNumber
2 of SDS asci
Map distance 100
Total number of asci
1.5.2 Analysis of unordered tetrad
Unordered tetrad contains group of spores that randomly arranged and the product of meiosis. Unordered
a are
tetrad analysis can be used to map genes in dihybrid crosses. This analysis can determine if two genes are linked
or assort independently. If two genes are linked, a tetrad analysis can also be used to compute the map distance.
Three patterns of segregation are possible in the tetrads when two pairs of alleles are segregating. For exampie,
in a cross ABxab, the three types of tetrad are:
Type 1: AB AB ab abb
If a crossover does not occur between these two loci or if a two-strand double crossover occurs between them, the
resulting meiotic products will be of two kinds, resembling the parental combinations. Such a tetrad is referred to
as parental ditype or PD.
No crossover
d
a
a b
b
B Two strand double
X O
crossover
Figure 1.26 Parental ditypes.
Type 2: Ab Ab aB aB
two kinds of products, neither of which are parental
between the two genes results in
A four-strand double crossover
ditype or NPD.
non-parental
combinations. This tetrad is referred to as
Four strand double
A
crossover
a
a
Figure 1.27 Non-parental ditypes.
Type 3: AB Ab aB ab two genes result
three-strand double
crossover (of two types) between the
crossover or a
A two-strand single combinations. This tetrad is referred to
as tetratype or TT
and non-parental type
formation of both parental and 50% non-parental type).
are present (50% parental type
four of the possible
genotypes
40
Genetics
B
B
Two strand single
b crossover
BB a
b B
B
a
X Vb Three strand double
crossover
b
Figure 1.28 Tetratype.
To estimate the amount of recombination between the two
markers, we can use the formula:
NPD+ TT
Recombination frequency = x 100%
Total number of tetrads
