Mutations

A sudden heritable change

 Mutation is the sudden heritable change in the phenotype of an organism. In molecular term, mutation is the permanent and relatively rare change in the number or sequence of nucleotides which results due to change in in DNA bases in nuclear or cytoplasmic DNA or due to chromosomal aberrations.

 The term mutation was introduced by Hugo de Vries in 1900. Mutagenic action of X rays was discovered by Muller in 1927 on Drosophilla and of gamma rays and X-rays in 1928 by Stadler in barley. General characteristics of mutation (i) Mutations are generally recessive and dominant mutations also occur. (ii) Mutations are generally harmful to the organism. A small proportion of them (0.1%) are beneficial (iii)Mutations occur at random (i.e) they may occur in any gene or chromosome.

(iv) Mutations are recurrent (i.e) same gene may undergo mutations repeatedly. (v) Induced mutations often show pleiotropy due to close linkage of mutated gene with other genes. (vi) Macro mutations which occur in oligogenes are easily identifiable and micro mutations which occur in poly genes are not easily identifiable.
But some genes show higher mutation rates than others. Highly mutable sites are called mutational hot spots.

Change in allelic status is also classifies as below based on efficiency of the function of the mutated allelomorph. a) Hypomorph- allele functioning less efficient b) Hypermorph allele functioning more efficient c) Amorph- allele completely losing the function d) Neomorph- allele acquiring a new function e) Antimorph allele impacting the action of normal allele

Classification of mutations
1. Depending upon the size of genetic material involved in the mutation, mutations are classified as (i) Point mutation If the mutation affects a single gene causing a change in the allele status from dominant to recessive or vice-versa, the mutation is called as point mutation. It is also called as intra-genic mutation.

(ii) Chromosomal mutation If the mutation induces structural aberration in the chromosomes such as deletion, duplication, inversion and translocation (transversion) or number aberrations, the mutation is called as chromosomal mutation.
Change in allelic status may be from wild type to a new type called as forward mutation or may be from new type to wild type called as reverse mutation.

2. Depending upon the level of detectability of mutations / the magnitude of phenotypic effect produced. (i) Macro mutation If the mutation result in easily identifiable distinct morphological changes in the phenotype, the mutation is called as macro mutation. It is called as oligogenic mutation, as it is observed generally in qualitative characters which are controlled by oligogenes. Since, the phenotypic change is easily recognizable on individual plant basis; selection in M2 generation is easy.

(ii) Micro mutation If the mutation result is not easily identifiable and not clearly distinguishable morphological changes in the phenotype, the mutation is called as micro mutation. It is called as polygenic mutation as it is observed in quantitative characters which are controlled by polygenes. Micro mutations are of most economic value in plant breeding than macro mutation as most of the economic characters are governed by polygenes. Selection is carried out on M3 or later generations only.

3. Depending upon the survivability level of mutated individuals or the mutation effect on the survival level of individuals. i. Lethal: If the mutation kills all the individuals carrying the mutated gene. Dominant lethals (AA) do not survive in homozygous or heterozygous condition and hence cannot be studied. Recessive lethal (aa) do not survive in homozygous condition alone. ii. Sub-lethal: If the mutation kills more than 50% of the individuals Reduces the viability of the individuals to a greater iii. Sub-vital: If the mutation kills less than 50% of the individuals Sub-vital mutation reduces the viability of the individuals to a limited extent

iv. Vital: If the mutation does not kill any of the individuals Do not reduce the viability of the individuals Vital mutations occur in a much lower frequency (0.1% of all the mutations) as compared to other three types. Vast majority of mutations are lethal, sublethals, and sub-vital and are of little value in crop improvement Vital mutations are of high value in crop improvement

Molecular basis of mutations

In any organism, triplet codon consisting of three nitrogenous bases in the DNA codes for an amino acid and a specific combination of triplet codons produce several amino acids which unite through peptide linkage to produce a protein. Proteins (which include enzymes) decide the phenotype of the organism. Hence, any change in triplet codon (a single or no of bases) causes a change in the sequence of aminoacids in a protein result in the altered phenotype. A heritable change in N base is the ultimate cause of mutation. Chromosomal aberrations (change in no. or segment) occurring due to mutation also result in the change in the sequence of aminoacids or in the absence of entire protein.

 Depending upon the nature of change in the nitrogenous base of nucleotide DNA, molecular basis of mutation can be classified as below, A) Single base change at the particular point (i)Base substitution a) Transition b) Transversion (iii)Base addition (iv) Base deletion

B) Multiple base change at a particular point (i) Triplet base addition or deletion (ii) Non triplet base addition or deletion Frame shift mutation When a purine is replaced by another purine or a pyrimidine by another pyrimidine the base substitution is called as transition. When a purine is replaced by a pyrimidine or vice versa, the base substitution is called as transversion. Base addition When one or more nitrogenous bases in a DNA molecule is added.

Base deletion When one or more nitrogenous bases in a DNA molecule is lost. Frame shift mutation If the no. of bases added or lost is not a multiple of three, the sequences of all the triplet codons beyond the point of insertion or deletion are altered and all the codons code for a different amino acid. Thus the reading frame of the subsequent codons is shifted in such mutations. This type of mutation is called as frame shift mutation.

 A frame shift mutation, changes all the amino acids of the concerned protein, located subsequent to the addition or deletion of bases. In such cases, the concerned protein becomes non functional. Hence, such mutations are much more deleterious than those produced by base substitution except non-sense mutations.

Change in a base at atom level. Tautomerization: When the hydrogen atoms in a DNA molecule gets shifted from one position to another in a purine or in a pyrimidine, the process is called as tautomerization and the new product is called as tautomer.

Depending upon the type of cells involved in the mutation

(i) Germinal mutation Occurs in the generative cells (reproductive cells) of an organism, the mutation is called as gametic mutation and passed on to the next generation, via the gametes. (ii) Somatic mutation Occurs in the somatic cells of an organism (a) Non heritable somatic mutation In sexually reproducing organisms. (b) Heritable somatic mutation (Bud mutation) In asexually reproducing organisms, Such somatic mutations occurring in vegetative propagation is called as bud mutations. Since, in general buds are the core vegetative propagating part.

Depending upon the presence or absence of artificial causal factors

Spontaneous mutation If the mutation occurs naturally without the artificial treatment by physical or chemical agents, the mutation is called as spontaneous mutation. Generally frequency of spontaneous mutations is one in 10 lakhs. i.e 10-6. However, some loci undergoes spontaneous mutations at high frequency. e.g R locus in maize with a frequency of 4.92 x 10-4, while Wx locus is highly stable. Spontaneous mutation of certain genes are influenced by the genetic background in which it is present.

 Certain intrinsic and extrinsic factors produce spontaneous mutation. Genetic instability due to hybridity or polyploidy or due to specific genes Extrinsic (external) factors causing spontaneous mutation are nutrition, temperature, naturally occurring radiations are chemicals and very high oxygen pressure.

Induced mutation
If the mutation occurs due to the artificial treatment by physical or chemical mutagenic agents, the mutation is called as induced mutation. Induced mutation produces new allele similar to spontaneous mutations. But in contrast to spontaneous mutations, very high frequency of mutation can be induced through the mutagenic agents at desired level. Hence induced mutations are more useful in crop improvement. The method of crop improvement utilizing the induced mutation is known as mutation breeding. The agents that induce mutation are known as mutagens.

Mutagens Mutagens may be physical or chemical. I. Physical mutagens Physical mutagens are radiations. Radiation causing mutations are of two types
(1) Ionizing radiation (2) Non-ionizing radiation.

1. Ionizing radiation: Ionizing radiations are those which when pass through matter, transfer energy to the matter rendering it to lose electrons.

A. Non particulate (Electromagnetic and parsely

ionizing) X-rays- They are produced by X-ray machines. They are sparsely ionizing, non particulate and penetrating. Hard X-rays have wave lengths of 0.1 to 0.01A and soft x-rays have 1 to 10 A wave lengths. Gamma rays They are produced by 60Co and other radioactive isotopes. They are shorter in wave length than x-rays (0.01 A) and more penetrating than X-rays.

B. Particulate
(iii) Alpha particles: They are produced by radioisotopes of heavier elements. They have two protons and two neutrons. They are positively charged and less penetrating than neutrons and beta rays; Densly ionizing. (iv) Beta particles : They are high energy electrons produced by decay of radioactive isotopes like 32P, 35S, 3H. They are more penetrating than a particles, but less penetrating than X rays; Sparsely ionizing. (v) Fast and thermal neutrons: They are neutral in charge and produced by cyclotron of atomic reactors by radioactive decay of heavier elements. They are highly penetrating.

2. Non ionizing radiations Non ionizing radiations are those which when pass through matter transfer energy to the matter rendering its electrons to move to higher energy levels (higher orbits). Incase of ionizing radiations, electrons are lost by the matter, but incase of nonionizing radiations, electrons are raised to higher energy levels (Excitation). The atoms in excited state shows increased activity. e.g. UV rays: They are produced by mercury vapour lamp and the weave length is 100-3900A. They are lower in energy and less penetrating. Hence, thin tissues like pollen of plants or eggs of Drosophila are irradiated using UV rays to induce mutation. Since, DNA bases show maximum absorption at 540A.

 The wavelength of UV is suitable for mutation. Dimer formation and de-amination occur due to UV radiation. Chemical mutagens: Oehlkers in 1943 found that mixtures of ethylurethane and potassium chloride induced translocations in Oenothera. Auerbach and Robson reported the mutagenic action of mustard gas in 1946.

Different categories of chemical mutagenic agents are given below: 1. Alkaylating agents (i) Sulphur mustards: e.g. Dichloro Diethyl Sulphide (mustard gas) (ii) Nitrogen mustards: e.g. 2 Chloro ethyl dimethyl amine, Di (2chloro ethryl) amine (iii) Epoxides: e.g. Ethyl oxide, Glycidol (iv) Ethylene imines: e.g. Ethylne imine (EI), Acetyl ethylene imine (v) Sulphate, Sulphonates, Sultones and lactones:e.g. Dimethyl sulphate (DMS), Diethyl sulphate (DES), Methyl methane sulphonate (MMS), Ethyl methane sulphonate (EMS) CH3 SO2OC2H5) (vi) Diazo alkanes and nitroso compounds. e.g.Diazomethane, Diagoethane, Nitroso ethyl urea. 2. Acridine dyes eg. Acridine orange, Ethidium bromide.

3. Base analogues e.g. 5- Bromo Uracil, 5- Chloro Uracil, 2- Amino Purine Among these some of the alkylating agents viz, EMS, MMS, DES are frequently used for induced mutagenesis. Mechanisms of chemical mutagenesis i. Tautomeric shift: If the chemical mutagens change the amino group (-NH2) into imino group (- NH) in purines, A (amino adenine) = T pairing in DNA is converted to A (imino adenine) = C pairing. Similarly C=G pairing is to converted to C = A pairing. This is called as tautomeric shift. In case of pyrimidines, keto group (C=O) is converted to enol group (COH). Because of this A = T (keto) pairing in converted to G=T (enol) pairing.

(ii) Substitution: Base analogues substitute for purine and pyrimidines during nucleotide and DNA synthesis. (iii) Deamination: Amino group (NH2) is converted to keto group(C=O). Adenine is converted to hypoxanthine and cytosine is converted to uracil