12.

Mutagens
Artificial induction of mutations: Mutations can be induced artificially using

1. Physical mutagens or radiations

2. Chemical agents
Classification of mutagen
1. Physical mutagens: Include various types of radiations, viz., x-rays, γ-rays, a-
rays, ß-rays, fast neutrons, thermal or slow neutrons, UV rays etc. The physical
mutagens are classified into
a) Ionizing radiations: They work through the release of ions. They have
deep penetrating capacity. Eg: x-rays, γ-rays, a -particles etc.
b) Non-ionizing radiations: They function through excitation and have a
very low penetrating capacity. They are used for studies on bacteria and
viruses. Eg. UV rays.

Sources of physical mutagens:

 Gamma garden
 Gamma green house
 Vertical gamma irradiation facility
 Horizontal gamma irradiation facility
 X-ray machine
 Isotopes
 Small portable irradiators, accelerators and cyclotrons
 Nuclear reactors

2. Chemical mutagens: These can be divided into four groups.

a) Alkylating agents: This is the most powerful group of mutagens. These are
the chemicals which are mainly used to induce mutations in cultivated plants.
They induce mutations especially transitions and transversions by adding an
alkyl group (either ethyl or methyl) at various positions in DNA. Alkylation
produces mutation by changing hydrogen bonding in various ways. Eg:
Dimethyl sulphonate (DMS), Ethyl methane sulphonate (EMS), Nitrosomethyl
Urea (NMU), Nitrosoethyl Urea (NEU), Methyl methane sulphonate (MMS).
b) Base analogues: These are chemicals which are very similar to DNA bases,
such chemicals are sometimes incorporated in DNA in place of normal bases
during replication. Thus, they can cause mutation by wrong base pairing. An
incorrect base pairing results in transitions or transversions after DNA
replication. Eg: 5– bromouracil, 3-bromodeoxy uridine, 2-amino purine.
c) Antibiotics: A number of antibiotics like mitomycin and streptomycin have
been found to possess chromosome breaking properties. Their usefulness for
practical purposes is very limited.
 d) Acridine dyes: Acridine dyes Eg: proflavin, acriflavin, acridine orange, etc.
are very effective mutagens. These are positively charged and they insert
themselves between two base pairs of DNA. This is known as intercalation.
Replication of intercalated DNA molecules results in addition or deletion of
one or few base pairs which produces frame shift mutations.
e) Deaminating agents: deaminating agents has strong mutagenic activity in a
variety of viruses and microorganisms. But not useful in higher plants.
f) Miscellaneous: Hydoxyl amine produce chromosomal aberrations. Nitrous
acid.

Materials used for treating with mutagens:

Seeds, pollen, vegetative buds, whole plants, bulbils, tubers, suckers etc.
Molecular basis of mutations:
The term mutation is presently used to cover only those changes which alter the
chemical structure of the gene at molecular level. Suchchanges are commonly referred
to as “point mutations”. Point mutations involve a change in the base sequence of a
gene which results in the production of a mutant phenotype. Point mutations can be
subdivided into the following three classes based on molecular change associated with
them.
1. Base substitution
2. Base deletion
3. Base addition
1. Base substitution: When a single base in a DNA molecule is replaced by another
base it is known as base substitution (see Fig.). This can be of two types.
a) Transition: Replacement of a purine by another purine or a pyrimidine by
another pyrimidine. (or) The substitution of a purine by another purine or of a
pyrimidine by another pyrimidine base in DNA or RNA is known as
transition.

(A G or C T)

b) Transversion: Replacement of a purine by a purimidine and vice versa. (or)

The substitution of a purine by a pyrimidine or of a pyrimidine by a purine in
DNA or RNA is known as transversion.

(A or G C or T or U)
A base substitution may cause Neutral, Sense (silent), Missense or Nonsense
mutation (see Fig.)

A. Neutral mutation: A change in a base pair results in an amino acid change

but the new amino acid has the same chemical properties as the old amino
acid.
B. Sense (silent) mutation: when one nucleotide is changed to another
nucleotide but it doesn’t affect the amino acid (protein) the gene codes for. In
iother word a change in base doesn’t significantly alter the phenotype of the
organism in which they occur.
C. Missense mutation: change in a base produced altered protein.
D. Nonsense mutation: change in one base leads results in a premature stop
codon, or a nonsense codon which yield incomplete, and usually non-
functional protein product
2. Base deletion: In base deletion, one or more bases are altogether deleted
3. Base addition: There is insertion of one or more bases.

Addition or loss of bases in multiple of three, added or delete one to several amino
acids from concerned protein, which may or may not give profound effect on activity
of polypeptide. But if the number of bases added or deleted is not a multiple of three,
a frameshift mutation is obtained, as the reading frame in such case is shifted from the
point of addition or deletion onwards. Hence, in a frameshift mutation, all the amino
acids of a polypeptide chain located beyond the site of mutation are substituted /
altered.

Frameshift mutations: The mutations which arise due to addition or deletion of

nucleotides in mRNA are known as frameshift mutations, because the reading
frame of base triplets (codons) beyond the point of addition or deletion is altered as a
consequence of such mutations (see Fig.).

Detection of sex linked lethal mutations in Drosophila by Muller’s ClB technique

Muller developed a system, ClB technique for detecting recessive sex-linked
lethal mutations induced by X-ray treatment in Drosophila. He used a heterozygous
(ClB) stock of Drosophila, which has a special X-chromosome, a large part of
which is inverted (paracentric inversion).This acts as a crossover suppressor in the
inverted region and is designated by C. A recessive lethal (l) gene and the
dominant gene for bar (B) eye are located within the inverted segment, as a
result, the l and B are always inherited together in the same chromosome. The
other X-chromosome was normal. Consequently, all ClB females identified by the
bar eye shape, are heterozygous for this chromosome, while the males having the ClB
chromosome do not survive [because of the l (lethal) gene].

The male flies were irradiated with X-rays for the induction of sex-linked
recessive lethal mutations. Such males are crossed with ClB females. In the F 1 half
of the females will have the ClB chromosome, which are easily identified by the bar-
shaped eyes. The remaining half of the females will not have the ClB chromosome
and are rejected. All the surviving F1 males will have the normal chromosome from
the ClB females, while those receiving the ClB chromosome will die due to the lethal
gene ‘l’. Each F1 ClB females is mated to a normal male. Progeny from each such
mating is kept in separate culture bottles. Each F 1 ClB female will have one ClB X-
chromosome and one X-chromosome from the mutagen treated male parent. So, half
of the male progeny receiving ClB X-chromosome will die. The remaining half male
progeny will receive their X-chromosome from their mutagen treated grandfather
which may or may not carry the induced mutation. In case lethalmutation was
induced, no males will be observed. On the other hand, if no lethal mutation was
induced, half of the males will survive. Thus, the ClB method was the simple, rapid
and most efficient method for detecting sex-linked lethal mutations.

Detection of mutations in plants:

Generally, the seeds of a variety or strain are treated with the mutagens and
grown to obtain M1 plants. The M1 plants are selfed to avoid out crossing due to
partial male sterility in M1 plants. The seeds thus obtained represent the M2
generation. The M2 plants are grown and the plants having mutant features are
counted. Then the frequency of a given mutation is estimated as per cent ratio
between the number of plants exhibiting a mutant phenotype in M2 and the total
number of plants in M2 .

No. of plants exhibiting mutant phenotype in M 2

Mutation frequency (%) = x 100
Total number of plants in M 2
Significance of mutations in Plant Breeding:
1. When a variety is exceptionally good except for one or few characters.
 2. When a recessive character is desirable and transfer of that character from wild
species is difficult.
3. When a desirable character is linked with an undesirable character. 4. If there is
no known source of resistance gene in the available germplasm
4. To create variability
5. To develop male sterile lines
6. To create variations in vegetatively propagated plants

Chimeras
Chimera is defined as a mixture of genetically diverse tissues in the same shoot.
These tissues frequently form mosaic pattern. The most common and easily observed
chimeras are leaf variegations observed in horticultural plants like Codium and
Acalypha. The leaf variegations or chlorophyll variegations are due to plastid
mutations, cons isting of green and white or green and yellow patterning as also the
patterning of anthocyanin distribution. These chimeras can be of three types.

1. Sectorial: The tissues possess the characteristics which closely resemble the
parents from which these tissues are originally derived.
2. Periclinal: The core is of one plant and the epidermal region is of another plant.
As a result, the leaves may be of one plant type and the flowers and fruits are of
another plant.
3. Chromosomal chimera: The most frequent being the ploid-chimera. It has
been a very common experience to meet with cells especially in the root tips,
with higher chromosomal number in an otherwise diploid tissue. Plants under
experimental control have been sometimes observed to give rise suddenly to
sectors or whole branches differing in chromosome number.

Xenia
Effect of the genotype of pollen grain on the phenotype of seed tissues (embryo and
endosperm) or the genetic effect of pollen parent upon the embryo and endosperm of
seeds in some plants.