Principles of Genetics 2+1

Theory

Unit I: Cytology
Definition of genetics, heredity, inheritance, cytology, cytogenetics; Brief history of
developments in genetics and cytogenetics; Physical basis of heredity: Structure and function of
cell and cell organelles – Differences between Prokaryotes and Eukaryotes.Cell division – mitosis,
meiosis and their significance, cell cycle - zygote formation and embryo development - identical
and fraternal twins. Chromosome structure, chemical composition, nucleosome, centromere,
telomere, euchromatin, heterochromatin, NOR, satellite chromosome, karyotype, ideogram –
chromosome banding; Types of chromosomes based on position of centromere, based on structure
and function: normal and special chromosomes - polytene, lampbrush, based on the role in sex
determination: autosomes and allosomes, Other types of chromosomes - B, ring and
isochromosomes; Chromosomal aberration: Variation in chromosome structure – deletion,
duplication, inversion and translocation – genetic and cytological implications; Chromosomal
aberration: Variation in chromosome number – euploid, aneuploid, types of aneuploids and their
origin; Nondisjunction - Klinefelter syndrome and Turner syndrome; Definition of eugenics and
euthenics; Polyploid - auto and allopolyploids, their characters; meaning of genome; evolution of
wheat, Triticale, cotton, tobacco, Brassicas.

Unit II: Mendelian laws and modifications of Mendelian laws

Pre-Mendelian ideas about heredity – Vapour and fluid theory, Magnetic power theory,
Preformation theory, Lamarck’s theory, Darwin’s theory, Germplasm theory and Mutation theory;
Work of Mendel – Characters studied reasons for Mendel’s success, Law of dominance, Law of
segregation and Law of independent assortment. Rediscovery of Mendel’s work; Chromosomal
theory of inheritance. Allelic interactions – Dominance vs. recessive, complete dominance,
codominance, incomplete dominance, over dominance; Terminologies: gene, allele, locus,
homozygous, heterozygous, hemizygous, genotype, phenotype, monohybrid, dihybrid, trihybrid,
polyhybrid. Deviation from Mendelian inheritance – Non allelic interaction without modification
in Mendelian ratio – Batson and Punnet’s experiment on fowl comb shape. Non allelic interaction
with modification in Mendelian ratio – i.) Dominant epistasis (12:3:1) ii.) Recessive
epistasis(9:3:4) iii.) Duplicate and additive epistasis((9:6:1). iv.) Duplicate dominant
epistasis(15:1) v) Duplicate recessive epistasis (9:7) vi.) Dominant and recessive epistasis(13:3);
Summary of epistatic ratios (i)to (vi); Lethal genes, Pleiotrophy, penetrance and expressivity,
phenocopy: Multiple alleles, blood group in humans, coat colour in rabbits, self incompatibility in
plants; pseudo alleles, isoalleles;

Unit III: Quantitative inheritance, Linkage and Crossing over

Quantitative inheritance – Multiple factor hypothesis – Nilsson Ehle, his experiment on
wheat kernel colour; Polygenes – transgressive segregation, comparison of quantitatively and
qualitatively inherited characters; modifiers; Types of gene action controlling quantitative traits;
Linkage - coupling and repulsion; Experiment on Bateson and Punnet – Chromosomal theory of
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linkage of Morgan – Complete and incomplete linkage, Linkage group; Crossing over –
significance of crossing over; cytological proof for crossing over - Stern’s experiment; Factors

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controlling crossing over; Strength of linkage and recombination; Two point and three point test
cross; Double cross over, interference and coincidence; genetic map, physical map.

Unit IV: Sex determination, sex linkage and cytoplasmic inheritance

Sex determination: Autosomes and sex chromosomes - chromosomal theory of sex

determination - different types – sex determination in human, fowl, butterfly, grasshopper, honey
bee, fumea; Genic balance theory of Bridges, quantitative theory, hormonal theory, barr bodies,
metabolic differentiation theory; Gynandromorphs – sex reversal in chicken; Sex linked
inheritance – cris cross inheritance – reciprocal difference; holandric genes; sex influenced and
sex limited inheritance; Sex determination in plants – Melandrium, papaya, maize. Cytoplasmic
inheritance and maternal effects – features of cytoplasmic inheritance, chloroplast, mitochondrial
- plastid colour in Mirabilis jalapa - iojap gene of maize, cytoplasmic male sterility in rice, kappa
particles of paramecium - plasmid and episomic inheritance.

Unit V: Modern concept of genetics and mutation

DNA, the genetic material – Griffith’s experiment, experiment of Avery, McCleod and
McCarthy – confirmation by Hershey and Chase; RNA as genetic material – Frankel, Conrat and
Singer experiment; Structure of DNA – Watson and Crick model – Central dogma of life; Proof
for semi conservative method of DNA replication; Models of DNA replication; steps involved in
DNA replication; RNA types - mRNA, tRNA, rRNA; genetic code, transcription; Translation –
protein synthesis; Regulation of gene expression – operon model of Jacob and Monad; Structural
genes and regulator genes; Cistron, muton and recon; Complementation test; exons, introns – split
genes – plant genome structure; Mobile genetic elements; Meaning of Developmental genetics,
DNA methylation, siRNA, RNAi, Functional genomics, Metagenomics, Transcriptomics,
Proteomics, Metabolomics and Phenomics. Mutation – characteristics of mutation – micro and
macro mutation – ClB technique - molecular basis of mutation; major physical and chemical
mutagens.

Practical
Study of cell and cell organelles – Preparation of fixatives and stains – pre treatment of
materials for mitosis and meiosis – study of mitosis and meiosis. Study of genetic ratios of –
monohybrid, dihybrid – incomplete dominance. Gene interaction - multiple alleles and multiple
factors. Study of linkage, estimation of strength of linkage and recombination frequency in two
point and three point test cross data and F2 data – Drawing of genetic map – interference and
coincidence.

Theory schedule
1. Definition of genetics, heredity, inheritance, cytology, cytogenetics; Brief history of
developments in genetics and cytogenetics.
2. Physical basis of heredity: Structure and function of cell and cell organelles – Differences
between Prokaryotes and Eukaryotes.

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3. Cell division – mitosis, meiosis and their significance, cell cycle; zygote formation and embryo
development - identical and fraternal twins.
4. Chromosome structure, chemical composition, nucleosome, centromere, telomere,
euchromatin, heterochromatin, NOR, satellite chromosome, karyotype, ideogram –
chromosome banding.
5. Types of chromosomes based on position of centromere, based on structure and function:
normal and special chromosomes - polytene, lampbrush, based on the role in sex
determination: autosomes and allosomes, Other types of chromosomes - B, ring and
isochromosomes.
6. Chromosomal aberration: Variation in chromosome structure – deletion, duplication, inversion
and translocation – genetic and cytological implications.
7. Chromosomal aberration: Variation in chromosome number – euploid, aneuploid, types of
aneuploids and their origin; Nondisjunction - Klinefelter syndrome and Turner syndrome;
Definition of eugenics and euthenics.
8. Polyploid - auto and allopolyploids, their characters; meaning of genome; evolution of wheat,
Triticale, cotton, tobacco, Brassicas,
9. Pre-Mendelian ideas about heredity – Vapour and fluid theory, Magnetic power theory,
Preformation theory, Lamarck’s theory, Darwin’s theory, Germplasm theory and Mutation
theory.
10. Work of Mendel – Characters studied reasons for Mendel’s success, Law of dominance, Law
of segregation and Law of independent assortment. Rediscovery of Mendel’s work
11. Chromosomal theory of inheritance. Allelic interactions – Dominance vs. recessive, complete
dominance, codominance, incomplete dominance, over dominance.
12. Terminologies: gene, allele, locus, homozygous, heterozygous, hemizygous, genotype,
phenotype, monohybrid, dihybrid, trihybrid, polyhybrid.
13. Deviation from Mendelian inheritance – Non allelic interaction without modification in
Mendelian ratio – Batson and Punnet’s experiment on fowl comb shape. Non allelic interaction
with modification in Mendelian ratio – i.) Dominant epistasis (12:3:1)
14. ii.) Recessive epistasis(9:3:4) iii.) Duplicate and additive epistasis((9:6:1). iv.) Duplicate
dominant epistasis(15:1)
15. v) Duplicate recessive epistasis (9:7) vi.) Dominant and recessive epistasis(13:3); Summary of
epistatic ratios (i)to (vi).
16. Lethal genes, Pleiotrophy, penetrance and expressivity, phenocopy: Multiple alleles, blood
group in humans, coat colour in rabbits, self incompatibility in plants; pseudo alleles, isoalleles.
17. Mid Semester Examination
18. Quantitative inheritance – Multiple factor hypothesis – Nilsson Ehle, his experiment on
wheat kernel colour.
19. Polygenes – transgressive segregation, comparison of quantitatively and qualitatively
inherited characters; modifiers; Types of gene action controlling quantitative traits.
20. Linkage - coupling and repulsion; Experiment on Bateson and Punnet – Chromosomal theory
of linkage of Morgan – Complete and incomplete linkage, Linkage group.
21. Crossing over – significance of crossing over; cytological proof for crossing over - Stern’s
experiment; Factors controlling crossing over.
22. Strength of linkage and recombination; Two point and three point test cross.
23. Double cross over, interference and coincidence; genetic map, physical map.

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24. Sex determination: Autosomes and sex chromosomes - chromosomal theory of sex
determination - different types – sex determination in human, fowl, butterfly, grasshopper,
honey bee, fumea; Genic balance theory of Bridges, quantitative theory, hormonal theory, barr
bodies, metabolic differentiation theory; Gynandromorphs – sex reversal in chicken
25. Sex linked inheritance – cris cross inheritance – reciprocal difference; holandric genes; sex
influenced and sex limited inheritance.
26. Sex determination in plants – Melandrium, papaya, maize.
27. Cytoplasmic inheritance and maternal effects – features of cytoplasmic inheritance,
chloroplast, mitochondrial - plastid colour in Mirabilis jalapa - iojap gene of maize,
cytoplasmic male sterility in rice, kappa particles of paramecium - plasmid and episomic
inheritance.
28. DNA, the genetic material – Griffith’s experiment, experiment of Avery, McCleod and
McCarthy – confirmation by Hershey and Chase; RNA as genetic material – Frankel, Conrat
and Singer experiment.
29. Structure of DNA – Watson and Crick model – Central dogma of life
30. Proof for semi conservative method of DNA replication; Models of DNA replication; steps
involved in DNA replication.
31. RNA types - mRNA, tRNA, rRNA; genetic code, transcription.
32. Translation – protein synthesis; Regulation of gene expression – operon model of Jacob and
Monad; Structural genes and regulator genes;
33. Cistron, muton and recon; Complementation test; exons, introns – split genes – plant genome
structure; Mobile genetic elements; Meaning of Developmental genetics, , DNA methylation,
siRNA, RNAi, Functional genomics, Metagenomics, Transcriptomics, Proteomics,
Metabolomics and Phenomics.
34. Mutation – characteristics of mutation – micro and macro mutation – ClB technique -
molecular basis of mutation; major physical and chemical mutagens.

Practical schedule
1. Use of microscopes and study of cell shapes and cell organelles of active mitotic and meiotic
tissues.
2. Principles of killing and fixing; preparation of stains and preservatives.
3. Study of the mitotic phases in root tips of onion / Aloe sp.
4. Study of behaviour of chromosomes in mitosis.
5. Procedure for fixing and observing different meiotic phases in the inflorescence of maize.
6. Procedure for fixing and observing different meiotic phases in the inflorescence in pearl
millet/ sorghum/ forest tree.
7. Observation of bivalents, trivalents, quadrivalents and chromosome banding
8. Repetition of meiotic studies in maize/ sorghum/ pearl millet/ forest tree and making temporary
and permanent slides.
9. Principles of dominance, recessive, back cross, test cross, incomplete dominance,
codominance and lethal factor; Chi square test; Monohybrid genetic ratio with dominance, with
incomplete dominance and test cross.
10. Dihybrid ratio with dominance, with incomplete dominance and test cross
11. Simple interaction of genes-comb character in fowls; Dominant epistasis.
12. Recessive epistasis, Duplicate and additive epistasis.
13. Duplicate dominant epistasis, Duplicate recessive epistasis, Dominant and recessive epistasis.
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14. Multiple alleles and polygenic inheritance
15. Estimation of linkage with F2 and test cross data; Coupling and repulsion.
16. Problems on two point test cross and three point test cross; Working out interference,
coincidence and drawing genetic maps.
17. Final Practical examination.

References
1. Gupta P.K., 1997. Cytogenetics. Rastogi Publications, Meerut
2. Strickberger. M.W. 1996. Genetics. Prentice-Hall of India Pvt. Ltd. New Delhi.
3. Singh, B.D. 2004. Fundamentals of genetics, Kalyani Publishers, Chennai.
4. Verma,P.S. and V.K.Agarwal. 2007. Genetics. S.Chand and Company Ltd./ New Delhi.
5. Stansfield, W.D.1990. Theory and problems of genetics. Mc-Graw Hill Book Co.,New York

Further reading
1. Daniel Sundararaj, G. Thulasidas and M.Stephen Dorairaj, 1997. Introduction to
Cytogenetics and Plant Breeding. Popular Book Depot, Chennai –15.
2. Benjamin Lewin 2005 Genes IX Oxford University Press, Oxford.
3. Gupta P.K., 1993. Genetics, Rastogi publications, Meerut.
4. Reddi, O.S., 1992. Understanding Genetics. Sunil Sachdev Publishers, New Delhi – 64.
5. Russel, P.J. 2000. Fundamentals of genetics. Addition Wesley Longman Publishers, USA.
6. singh, R.J.2002. Plant cytogenetics. CRC Press, USA

Web resources
1. www.nmsu.edu
2. www.biology200.gsu.edu

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Lecture 1. Definition of genetics, heredity, inheritance, cytology, cytogenetics; Brief history
of developments in genetics and cytogenetics.

GENETICS
Genetics is the science of heredity and variation. The science of genetics deals with the
principles that explain the similarities between parents and their progeny and the differences
among individuals of a single species.
Heredity - Process which brings about the biological similarity between parents and progeny.
Deals with inheritance of characters from parents to off springs.
Inheritance is the transmission of genetic information from parents and ancestors to offspring.
Variation: The differences among individuals of a single species for a particular character.
Genes: are the functional units that govern the development of characters of an individual.
Gene - unit of inheritance
Cytology: Study of cell structure and functions of cell organelles

Cytogenetics -The study of various aspects of chromosomes and their effects on the development
of characters of organism.

Chief Events in Cytology

Year Scientist Discovery/ Event

1665 Robert Hooke discovered Cell
1831 Robert brown presence of nucleus
1838 Schleiden and Schwann Cell theory
1861 Schultzee Protoplasm theory
1870 Fredrick Meischer Isolated Nucleoprotein
1879 Flemming described chromatin in nucleus
1882 Flemming described cell division (Mitosis)
1888 Waldeyer Described chromosomes
1902 Mc Clung Sex Chromosomes
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 1903 Sutton Chromosome theory
1905 Farmer and Moore Coined the term Meiosis
1913 Sturtvent Built first chromosome map
1937 Blakeslee Induced polyploidy with Colchicine

Robert Hook (1665)

He described the cell as empty vessel. He introduced term cell.

Cameraious:
He proved pollen is important for fertilizations. He is the first man to produce first
artificial hybrid plant.
Koelreutes
He showed that F1 might resemble either male (or) female parents (or) combination of
both. Hereditary contribution of the two parents to their offspring was equal.
Knight
He obtained the dominant forms in F1 and segregation of various characters in F2.
Gaertner
F1 are uniform and their F2 produced considerable variation.
Naudin
Hybrids races and species of plants are often luxurient than either of the parents.
Robert Brown
He described the cell nucleus in the flowering plants.
He observed random thermal motion of small particles known as Brownian movement.
Schleiden and Schwann:
They discovered the formation of nucleus in the cell and formulated cell theory, which
says
1. The cell is the smallest building element of a multicellar organism.
2. Each cell has a specific work to complete.
3. The cell can only the produced from another by cell division.
Strasberger:
He described fertilization in Angiosperms.
Van Beneden - Meiosis:
He showed number of chromosomes in the gametes, is half of the number of body cells. In
fertilization the chromosome contribution of eggs and sperms to the zygote are numerically equal.
Flemming - Mitosis:
1. He proposed mitosis in cell.
2. He showed the chromosome spliited during nuclear division and the formation of
daughter nuclei.
3. He also applied the name chromatin which is to the stainable position of the nucleus.
History of Genetics
Gregor John Mendel
• An Austrian botanist who laid foundation for the science of genetics.

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 • Born in the year 1822 near Brunn in Austria
• He worked with Pisum sativum - Garden pea
• Presented a paper in 1865 – “Experiments in Plant Hybridization” before the Natural
History Society of Brunn
• Mendel’s Paper was published in 1866
• Formulated two important laws of inheritance in 1866
1. Law of segregation
2. Law of independent assortment
• Died in the year 1884
• Importance of his work was realized only in 1900
• For this pioneer work he was called as the "Father of Genetics"

Rediscovery of Mendels work in 1900 by Correns, Hugo devries, Tschermak

Carl Erich Correns
• A german botanist who rediscovered Mendel's work in 1900
• He conducted research with garden pea and came to the same conclusion as drawn by
Mendel in 1865.
• He worked with Mirabilis jalapa (4'O' clock plant) and established the first conclusive
example for Extrachromosomal inheritance
Hugo devries
• Rediscovered the mendel's law of inheritance independentantly but simultaneously with
Correns and Tschermak in 1900,
• He coined the term Mutation - sudden heritable changes in the characters
Tschermak
• One of the co discoverers of Mendel's classic papers on garden pea
• He applied mendel's law of heredity in barley, wheat rye hybrids and oat hybrids for
the development of new plants
Morgan, T.H.
• Established the chromosome theory of heredity in 1910.
• He showed that genes are linked in a series on chromosomes and are responsible for
observable genetic traits.
• Received Nobel Prize for Physiology and Medicine in 1933.
• Discovered hereditary transmission mechanisms and sex linkage in Drosophila.
Bridges, C.B.
• Established the chromosomal basis of heredity and sex.
• Constructed detailed gene map of the giant chromosomes found in the salivary gland
cells of fruit fly larva.
• Discovered genic balance theory of sex determination and gene duplication in
Drosophila.
Muller, H.J.
• X- rays speed up the natural process of mutation.
• For experimental induction of mutation he was awarded Nobel Prize
Beadle and Tatum
• Proposed the concept of one gene one enzyme hypothesis for which they received Nobel
prize in 1958

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 Avery, Mac Leod and Mc carty
• Discovered the phenomenon of transformation
• They (Avery, Mac Leod and Mc carty) reported that the substance which caused the
transformation was DNA and demonstrated the DNA was the genetic material
Watson and Crick (1953) and Wilkinson
• The X ray diffraction studies of DNA by Wilkinson resulted in the Discovery of the
molecular structure of DNA, in 1953 by Watson and Crick
Watson and Crick - proposed the double helix model of DNA
• The three scientists received Nobel prize in 1962
Barbara Mc Clintock
• Discovered transposons/ jumping genes in Maize in 1950- Genes that are capable of
changing their position on a chromosome and from one chromosome to another
• Awarded Nobel Prize in 1983

Benzer
• Detail structure of viral genes and coined the term cistron to denote functional sub unit
of genes
• Gave sub divisions of genes viz., Cistron, Recon and Muton. These are the units of
function, recombination and mutation within a gene.
Meselson and Stahl
• Experimentally confirmed the semi-conservative replication of DNA

F.H.C. Crick
• Proposed the Central Dogma of Molecular Biology in 1958
• Gave evidence for the triplet nature of genetic code in 1961
Nirenberg - Responsible for deciphering the genetic code- 1961

Jacob and Monad

• Explained gene regulation in cell metabolism by directing the biosynthesis of enzymes -
Operon concept in 1962
• Awarded Nobel prize in 1965
Khorana
• Discovered how the genetic components of the cell nucleus control the synthesis of
protein
• Awarded Nobel prize in 1968 with Nirenberg and Holley
• He prepared the first artificial copy of a yeast gene in 1970
Stanley Cohen and Herberd Boyer
• Genetic Engineering in 1973
A.M. Maxam and W.Frederick Gilbert; F. Sanger and Coulson
• Sequencing of DNA in 1977
Craig Venter (2001) – human Genome Project – Complete sequencing of human genome
Goff et al and Yu et al (2002) – Rice Genome Sequencing

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 Lecture 2. Physical basis of heredity: Structure and function of cell and cell organelles –
Differences between Prokaryotes and Eukaryotes.

CELL
A Cell may be defined as the structural and functional unit of a living being. It is the
minimal biological unit capable of maintaining and propagating itself. A study of the structural
and functional organization of different structures within a cell is known as 'Cytology'.
Cytogenesis concerns with the study of various aspects of chromosomes and their effects on the
development of characters of organisms. It is universally accepted that genes are located in
chromosome. Cytogenetics originated as a result of bringing two different branches of biology
namely cytology and genetics together.

HISTORY- the word 'Cell' has been derived from the Latin word Cellula meaning a small
compartment. The term was first used by Robert Hook (1665). Robert Hook who constructed the
first compound microscope observed the sections of Cork and opined that they contain honeycomb
like compartments. German biologists M.J. Schleiden and T.S. Schewann (1838) established the
'Cell theory' that all organisms are made up of cells. One of the significant discoveries of the cell
came from 'Robert Brown' (1830). He discovered the presence of a spherical body in the centre of
every cell, which he named 'Nucleus'. In 1835-37, Purkinje and Mohi independently discovered
that protoplasm is an important constituent of every cell and it
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plays an important role in every cell activity including division. Golgi (1838) discovered the golgi
apparatus, Balbiam (181) discovered chromosomes in the salivary glands of chironomus. At
amount the same time, Flemming (1882) studied cell division in detail and gave the name ' Mitosis'.
Endoplasmic reticulum was discovered by proter in 1945, while Benda gave the name
mitochondria to organells originally discovered by Hemming. Lyzosomes were discovered in 1955
by de Duve. The shape of cell may be variable like spherical, rectangular, flattened, oval, polygonal
triangular come like column etc., There is a great rang eof variation among cells in size also. This
small cell size can be encountered in coccus bacteria (0.2 to 0.5 m) while the largest size of the
cell is seen in Ostrich egg (Marly 15 cm).
Gross morphology of the cell
A generalized plant cell has an outer most envelope called the 'Cell wall'. This is absent in
animal cells. Internal to this in the plasma membrane. This encloses the nucleus and other
cytoplasmic inclusions suspended in cytoplasm. The inclusions are Ribosomes, Lysosomes,
Mitochondria, Plastids, Golgi complex, Endoplasmic reticulum, Vacuole and non-living inclusions
like crystals, raphids etc., The primitive organisms like certain bacteria blue green algae, the
nucleus is not properly organized hence such cells are called Prokaryotic, while in evolved
organisms, the nucleus is organized. Such cells are called Eukaryotic. The following are some of
the fundamental differences between eukaryotic and prokaryotic cells.
CELL WALL - Plant cells are surrounded by a non-living and rigid coat called a 'cell wall'. The
main functions of a cell wall are to provide plant cells a definite shape and mechanical support and
strength to tissue and organs. Cell wall has 3 distinct parts ;
1. Middle lamella
2. Primary cell wall
3. Secondary cell wall

Middle lamella - In plants, the wall of contiguous (immediate neighbour) cells are joined by
middle lamella, which is composed mainly of pectin. Secondary wall Middle lamella Primary wall
Plasmodesmata The pectin of middle lamella is most likely in the form of calcium (Ca++) and
Magnesium (Mg ++) salts. Adhesian of the walls of cotinguos cells in primarily dependant on the
presence of Ca ++ and Mg++ ions in the middle lamella a removal of these ions results in

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the separation of cells from each other. Pectin is readily hydrolysed by the enzyme pectinase as
well as by strong acids.
PRIMARY CELL WALL - is deposited after the formation of middle lamella and lies between
middlle lamella and plasma lemma. It is main constituent are hemi cellulose (53%) and cellulose
(30%). In addition it contains pectin (5%). Protein (5%) and lipid (7%).
SECONDARY CELL WALL - Is the last to be deposited and lies between cell wall and plasma
lemma in a cell, it is the inner most laayer of wall. It is composed of mainly of cellulose. The
cellulose microfibrils are relatively more closely packed and they are arranged more or less parallel
to each other. Several microfibrils associate to form a macrofibril, which is the structural unit of
secondary cell wall.
PLASMA LEMMA (PLASMA MEMBRANE)
The membrane enclosing cytoplasm of a cell is known as plasma lemma or plasma
membrane. It is composed of lipids and preoteins, the ratio between the two being quite variable
among different cell types. Three distinct layers are seen under electron microscope, two or three
are relatively dense and osmiphilic in nature; each of them is about 20oA thich. The two osmaphilic
layers are separated by a relatively light osmiphobic layer of about 35oA thickness. The three
layers together are known as 'Unit membrane"' this term coined by Robertson. The chief function
of plasma lemma in to regulate the movements of various molecules into and out of the cytoplasm.
In addition to the passive movement of molecules, some ions are transported across plasma lemma
by means of active transport.
CYTOPLASM
The substance, except nuclear, surrounded by the plasmalemma is known as 'Cytoplasm".
Electron microscope reveals a member of membraneous and other structures in the cytoplasm; the
portion of cytoplasm other than therse structure is known as 'hyaloplasm. Of the various structures
present in the cytoplasm, mitochondria and plastids contain DNA; as a result they are autonomous
to a limited degree. However, the remaining cytoplasmic structures do not contain DNA and they
are specified exclusively by nuclear genes.
The cytoplasm may contain the following structures -endoplasmic reticulum (ER),
ribosomes, Golgi bodies, Lysosomes, Sphersomes, Vacuoles, certioles (in animals only),
microtubrils, Mitochondira and plastids (in green plants only).

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ENDOPLASMIC RETICULUM (E.R)
The cytoplasm contains an extensive network of membrane-enclosed space; these space
along with the membranes enclosing them are known as E.R. It consists of 3 types of membrane-
enclosed elements.
1. Vesicles of 25-500 in m diameter
2. Tubules of 50-100 m m diameter
3. 40-50 m m thick cisterns of variable length and width.
The tubulus may or may not be extensivley branched, and the cisterns may ormmay not be
connected with each other. The ultastructure of E.R membrane in the same as that of a unit
membrane, that is, it has two osmiophilic layers separated by an osmiopholic layers. E.R is grouped
into two categories,
1. Smooth E.R.
2. Rough E.R.
In smooth E.R elements, both outer and inner surfaces are regular and smooth. In those
cells where little or no protein synthesis takes place, only smooth ER is found. The rough ER
elements, ther outer surfaces of membranes have a rough appearance due to the attachment of
ribosome’s on the outer surface. Rough ER is mainly composed of cisterns (membrane-enclosed
plate like elements) and is found in cells actively involved in protein synthesis. Smooth and rough
E.R change into each other as per the needs of cells.
Functions of ER
i. it provides the structural base for protein (rough ER), lipid, phopholipid synthesis.
ii. it proivides channel for the transport of materials synthesized in association with
ER to the various parts of cells and even outside the cells.
iii. it provides a controlled passage for the export of m.RNA moelecules from
nucleus to rought ER.
iv. Several enzyme molecules are embedded in the membranes of E.R.

RIBOSOMES
These are dense granular nucleoprotein structures occurring in cytoplasm, matrix of
mitochondria and chloroplasts. In many instances ribosomes are attached to the ER. Observed

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first in plants cells in 1953 by Robinson and Brown, while studying bean roots. Ranging in
diameter from 150 to 200Ao, they have RNA and protein in equal quantities. Ribisomes are
isolated by differential centrifugation depending on sedimentation co-efficient. The sedimentation
coefficient is expressed in terms of Sved berg units. The 'S' units are related with the size and
weight of the ribosome molecules.
TYPES
Two types of ribosome’s have been identified based on the sedimentation coefficient. If
the organelle is heavier, its sedimentation co-efficient is more. The two types are 70s ribosome’s
and 80s ribosome’s. Ribosome’s may occur singly as isolated units when they are called
'monosomes'. When they occur in clusters or groups, they are called 'polyribosomes'. The poly
ribosomes may have a sedimentation coefficient of 100s-600s. The number of ribosomes per cell
varies, it may be 10,000 (bacterial cell) or up to 10 million (eukaryotic cell). Ribosomes of
chloroplasts and mitochondria have their own protein synthesis.
They have sedimentation co-efficient of 55s with two sub units 40s and 30s.
ULTRA STRUCTURE
Ribosomes are oblate or spheroidal structures having two sub units (a large and a small).
The larger sub unit in dome likes and the smaller subunit is placed above like a cap. The 70 s
ribosome has two units 50 s and 30s.

FUNCTIONS
Ribosomes are the sites of protein synthesis. The polyribosomes serve as a plat form in
the assembly of amino acids brought together by specific to RNA from cytoplasm.
GOLGI COMPLEX
Described first by Camilio Golgi in 1890. Golgi complex found in plant cell is often referred to as
'Dictyosomes". Each golgi body consists of following parts;
1. Cisternae
2. Tubulus
3. Vesicles
4. Golgian vacuoler.

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Functions
i. Absorption of compounds
ii. Sites of enzyme production
iii. Sites of hormonal production
iv. Sits of protein storage
v. Formation of plant cell wall-by synthezing pectin, hemicellulose and cellulose microfibrils.
They also help in the formation of cell plate during mitosis.

PLASTIDS
These are living cytoplasmic inclusions found inmost of the plants. The plastids are of
three categories viz., chromoplasts, leucoplasts and chloroplasts.
Chromoplasts
They are pigmented plastids. The pigments are non-chlorophyllus like carotenes, yellow
xanthophylls, phycoerythrin etc.,
Leucoplasts
They are colorless plastids. They lack pigments and are usually present in cells which do
not receive direct light. Leucoplast may be seen in the storage leaves of onion. Leucoplasts that
store starch are called amyloplast, those that store oil are called elaioplasts and the ones storing
proteins are called protenoplasts.

Chloroplast
These are by far, the commonest and the most plastids. As the primary sties for trapping
and converting solar energy they are very vital for the existence of not only the green plants, bu
for the whole living world.
Chloroplasts have varied shape and varied size. Chloroplast of polyploid cells are generally
larger than in the diploid cells. They are uniformly distributed all over the cytoplasm, but in some
instances they cluster towards the nucleus. The concentration of chloroplasts will also depend on
light intensity.
Structure

16
 It has a covering of two membranes with an inner membrane space. These membranes are
smooth and there are no perforations or particles. The membranes are differentially permeable.
A section of chloroplasts reveals an intricate system of membranes enclosed in a granular
matrix. These membranes are called lamellae and the surrounding matrix-the stroma. In a sectional
view, the lamellae can be seen packed and there stacks are called thylakoids. In higher plants, the
thylakoids themselves form highly compact bundles called grana. Some thylakoids of granum
extend into the stroma and maintain contact with other grana. These are called stroma thylakoid or
stroma lamellae or inter grana. Ribosomes and RNA have also been isolated from the chloroplasts
indicating a machinery for protein synthesis. Some of the important pigments present in chloroplast
are chlorophylls, carotenoids, cytochromes etc.,
NUCLEUS
It is the most important organelle of the cell which regulates all its activities. It was discovered
by Robert Brown (1831). Most of the cells are uninucleate. It has the following parts;
1. Nuclear membrane
2. Karyolymph (Nuclear sap)
3. Chromonemata
4. Nucleolus
5. Endosperms
NUCLEAR MEMBRANE
It helps in effective communication between nucleus and cytoplasm. The
elements of E.R. contribute to the nuclear envelope during cell division. The nuclear membrane is
a double membrane with a number of pores called ‘Nucleopores'. The space between these two
membranes is called 'perinucelar space' or cisterna.
KARYOLYMPH (NUCLEAR SAP)
It is proteinaceous, but also has nucleic acids, enzymes and minerals. It is quite probable
that in plants the nuclear sap contributes to the spindle.
CHROMONEMATA
Enclosed in the karylymph and visible in the interphase nucleus are found a number of
fibrillar structures constituting a network called chromonemata or chromatrin fibrils. Some
coarsegranules are deposited on the chromatin net work. These are called chromocentres and

17
constitute the points of condensation of chromosomes. During cell division the chromatin network
brakes up into specific number of chromosomes. Two regions can be identified in the chromatin
material. These are heterochromatic region and euchromatic region. The heterochromatic region
stains darkly and shows numerous bead like structure called 'Chromomeres'. The heterochromatic
region has less DNA. This region is believed to the genetically and metabolically inert. The light
staining region of the chromatin is called the 'euchromatin region" This region contains more of
DNA and is supposed to be genetically active. NUCLEOLOUS
Nucleolus was first discovered by Fortana (1874). A sphearoidal body, situated either in
the central or peripheral position, the necleolous is supported to regulate the synthetic activity of
the nucleus. Usually 2 or more chromosomes are associated with the nucleus (this can be seed
during late prophase) and these are called nucleolar organisms as they play a role in re-appearance
of the nucleolus after cell division. The number of nucleoli per nucleus varies from one to 2 or 3.
Chemically the nucleoli are rich in RNA.
Functions
i. It is the active site of RNA synthesis
ii. It is the source of ribosomal RNA
iii. It produces precursors of ribosome’s

Lecture3. Cell division – mitosis, meiosis and their significance, cell cycle; zygote
formation and embryo development - identical and fraternal twins.

CELL DIVISION
MITOSIS
All cells originate through division of pre-existing cells. Bodies of all multicellular
organisms are derived from unicellular zygote through repeated divisions of zygote and the cells
derived through its division. The division of chromosomes and cytoplasm of a cell into daughter

18
cells is known as 'Cell division'. The cell that undergoes division is termed as 'parent cell', while
the cells derived from the division of a parent cell are known as daughter cells.
Functions of cell division
To produce two daughter cells, which are involved in the following;
i. Growth and development of somatic tissue of organism
ii. Regeneration of damaged tissues
iii. Production of new tissues
iv. Reproduction
v. Keeping the size of cells within a limited range.
Two types of cell division i. Mitosis ii. Meiosis
In addition, bacterial cells divide by fission (similar to mitosis). The various
events occurring in division may be grouped into
i. Karyokinesis - Division of chromosomes
ii. Cytokinesis - Division of cytoplasm
MITOSIS
It was first used by Fleming in 1882. In plants, mitosis is confined to the meristamatic
tissues of root and shoots tips, young leaves flower buds. On the basis for changes in the
morphology of nucleus and the chromosomes, the events in a mitotic cell division are grouped into
five stages;
i. Interphase
ii. Prophase
iii. Metaphase
iv. Anaphase
v. Telophase.
Interphase
In this stage of cell after the telophase of previous division and before onset of prophase of
the next one. During interphase, chromosomes are fully extended and uncoiled so that they do not
take-up sufficient stain. Interphase in the longest stage. In a cell undergoing mitosis every 24 hours
i.e. having a cell cycle of 24 hr., interphase may occupy upto 23 hours, while the division

19
or mitotic phase may take up onluy 1 hour. DNA replication occurs during the middle part of
interphase. This provides the basis for classifying interphase into three sub stages.
1. G1 (first gap)
2. S (Synthesis of DNA)
3. G2 (Second gap)
G1, G2 - Protein + RNA synthesis
S - DNA synthesis
M - Chromosome movement, division

PROPHASE
i. The appearance of define thread like structures in nuclear is the most important event of
prophase. In the beginning, chromosomes appear as a loose ball of thin wool. As prophase
proceeds, chromosomes become increasingly shorter and thicker due to increased condensation by
mid prophase; the two chromatids of each chromosome become visible. By the end of prophase
all the chromosomes become considerably shorter and thicker.
ii. During prophase nucleolus and nuclear membrane remain present.
Chromosome condensation that is decrease in length, with increased in thickness is mainly due to
the coiling of chromosomes.
The two sister chromatids of each chromosomes are coiled in relation to each other this is referred
to as relational coiling. This relational coiling is of two types.
i. Plectominic coiling - The two sister chromatids cannot separate from each other without the
chromosomes being rotated. It is happening during prophase of mitosis.
ii. Paranimic coiling - Sister chromatids are note twisted round each other, they are simply
slipped into those of other they can easily separated without rotating the chromosomes. This type
of coiling is found during meiotic prophase. The relational coiling between sister chromatids goes
on decreasing as the chromosomes become smaller, it disappears by late prophase.

METAPHASE
At the end of prophase four important events take place;
The main features of metaphase are

20
i. Absence of nucleolus
ii. Disappearance of nuclear membrane.
iii. Arrangement of chromosomes on the equatorial plate
iv. Shortest and thickest chromosomes (Condensation)
v. Coils are less in number and largest in diameter
vi. Presence of spindle apparatus
vii. Absence of relation coiling between sister chromatids.

ANAPHASE
The two sister chromatids of each chromosomes separate and migrate towards the opposite
poles of the cell. Anaphase begin when the centromeres of chromosomes appear to divide
longitudinally so that the sister chromatids separate from each other and ends with the reaching of
the chromosomes to opposite poles centromete in the first portion of each of the chromosomes to
begin to move towards the poles. Spindle fibres originate at two points located near the periphery
of a cell and opposite to each other. These points are known as 'poles' Chromosomes become
somewhat more condensed as compared to those at metaphase, so that they appear relatively
smaller in size.

TELOPHASE
Anaphase ends and Telophase begins when sister chromatids of all the chromosomes of a cell
reach the opposite poles. During telophase, the following events occur in the two groups of
chromosomes collected at the opposite poles.
i. The chromosomes uncoil so that they become very long and thin and appeared to be coiled into
a loose ball of fine thread.
ii. Nucleus reappears
iii. Nuclear membrane is reorganized around each group of chromosomes.
iv. At the end of telophase, middle lamella appears at the equatorial plate of the cell.
The nuclear envelope dissociates into small elements which become part of E.R. of the cell. During
telophase, there elements re-originate around the two groups of chromosomes and fuse to produce
nuclear envelope around them. In terms of duration, prophase in the longest stage of the

21
division phase of cell cycle. In comparison anaphase is the shortest stage, while metaphase and
telophase are considerably longer than anaphase.

CYTOKINESIS
It is complete by the end of Telophase. At the equatorial plate, elements of E.R. and products of
Golgi bodies organize and give rise to cell plate and subsequently of cytoplasm begin in the centre
of the cell and gradually extend outwards on each side in a plane, perpendicular to the axis of the
spindle. The two daughter cells produced by mitosis contain one nuclear sac; each nuclear has the
same number of chromosomes as the parent cell. Each daughter cell enlarges in size till it becomes
comparable to the parent cell.

Significance of Mitosis
• After fusion of male and female gametes the zygotes are formed. So, Mitosis is
responsible for development of zygote into adult organism.
• it is essential for normal growth and development of living organism. It gives shape to a
specific organism.
• Mitosis, in plants leads to formation of new parts – roots, leaves, stem branches. It helps
in repairing of damaged parts.
• Mitosis, leads to production of identical progenies in vegetatively propagated crops.
• It is useful in maintaining purity of types because it leads to production of identical
daughter cells and does not allow segregation and recombination to occur.
• In animals, it helps in continuous replacement of old tissues. Eg. Blood cells.

MEIOSIS
Meiosis takes place during gamete formation and hence it is confined to reproductive cells only.
As a consequence of meiosis, gametes contain only half (h) of the somatic chromosome number
(2n). Therefore union between one male and one female gamete during fertilization restorers the
chromosome number to the diploid (2n) state. Thus the chromosome number of a species remains
constant from one generation to the next generation produced by sexual reproduction. In the
absence of meiotic cell division, the chromosome number of a species would be doubled in

22
every generation, due to the fusion of male and female gametes, an impossible biological situation.
The nucleus of each cell undergoes two successive divisions referred to as the first and second
meiotic division.

Pre-Meiotic Interphase
During 'S' phase of pre-meiotic interphase chromosomes replication takes place. But
approximately 0.3% of the total DNA present in the nucleus does not replicate during the 'S' phase
this DNA replicates during the zygotine sub stage of prophase I. A special type of histone specific
to cells preparing for meiosis is synthesized during S phase. This histone is not found in cells
undergoing mitosis, and it may be related to the entry of cells into meiosis.

FIRST MEIOTIC DIVISION

Significant events;
i. Pairing between homologous chromosomes.
ii. Crossing over between them during pachytene stage of prophase I
iii. Separation of homologus chromosomes and their migration to the opposite
poles of a cell during Anaphase I. As a result, the two daughter nuclei produced by this division
receive only half of the chromosomes present in somatic cells. For this reason, the first division is
often referred as 'Reduction division'.
Prophase I - is divided into 5 sub stage viz.,
i. Leptotene
ii. Zygotene
iii. Pachytene
iv. Diplotene
v. Diakinesis
LEPTOTENE
i. There is a marked increased in the nuclear volume
ii. There is chromosome condensation so that they become visible as fine
threads like a loose ball of knitting wool. Each chromosome consists of
two chromatids.

23
ZYGOTENE
It begins with the initiation of pairing between homologous chromosomes. The main
events are as follows:
i. Paring between homologous chromosomes.
ii. Completion of replication of the remaining 0.3% DNA of each nucleus, this DNA synthesis is
referred to as Z-DNA synthesis or Zygote DNA synthesis.
iii. Synthesis of a specific nuclear protein
iv. Development of the synaptenemal complex and
v. Progressive condensation of chromosomes.
Pairing of homologous chromosomes is often referred as 'Synapsis'.

PACHYTENE
It begins when synapsis comes to an end and it ends when the homologous chromosomes
begin to move away from each other. The main events are ; i. Thre is a further condensation of
chromosomes, so that chromosomes pairs become shorter and thicker. ii. Chromosomes are easily
recognizable during this stage and each bivalent has four chromatids. iii. The nucleolus is distinct
and quite large. iv. Crossing over between homologous chromosomes takes place during this
stage.

DIPLOTENE
i. Homologous chromosomes of each bivalent begin to move away from each other.
ii. The two homologous of each bivalent appear to be attached with each other at one or more
points, these attachments are known as chiasma. It is believed that initially chiasma are located at
the points of actual crossing over between homologous chromosomes.
iii. As diplotene progress, chiasmata, slowly move towards the ends of the homologous
chromosomes; this movement is referred to as chiasma terminalization i.e. movement of chaisma
towards terminal positions in the chrommosomes. Chiasma terminalization occurs mainly due to
the movement of homologous chromosome away from each other.
iv. There is further condensation of chromosomes so that they become progressively shorter and
thicker.

24
DIAKINESIS
i. Bivalents move away from each other and spread towards the periphery cells.
ii. Nucleolus, nuclear envelope disappears.
iii. The spindle apparatus is organized. The bivalents now migrate to the equatorial plate of cells;
this marks the ends of diakinesis. Bivalents may be in the form of (1) a closed ring, (2) an open
ring or (3) rod shaped.

METAPHASE -I
i. Bivalents are arranged at the metaphase plate
ii. Centomeres of the two homologues of each bivalent lie on the either side of the equatorial
plate.
iii. Metaphase terminates as soon as homologous chromosomes begin to separate from each other
and to migrate to opposite poles of the cell.

ANAPHASE -I
i. Separation of the two homologous chromosomes of each bivalent marks the beginning of
anaphase stage.
ii. One chromosome from each bivalent begins to migrate to one pole, while the other migrates to
the opposite pole.
As a result the number of chromosomes at each pole is exactly half (h) and each pole
receives one homologue from each of the bivalents present in a cell. Thus the reduction in
chromosomes number is not only a quantitative one but a qualitative one as well. Thus at the end
of AI, the chromosome present is somatic cells are effectively and precisely separated into two
identical groups.

TELOPHASE -I
i. The chromosomes uncoil only partially
ii. Nuclear envelope becomes organized around the two groups of chromosomes.
iii. Nucleolus also reappears.
CYTOKINENSIS

25
 The cytoplasm of each cell divides into two halves, with a single haploid nuclear in each
half. The two halves of each cell do not separate, but they stay together, and this two celled
structure is known as dyad.

SECOND MEIOTIC DIVISION / MEIOSIS II

During Meiosis II, two sister chromatids of each chromosome separate and migrate to the opposite
pole. As a result, the number of chromosomes in each of the two haploid nuclei remains the same
(i.e haploid)., at the end of this division. The second division of meiosis is often referred to as
equational division. Sometimes, it is called as 'Meiotic Mitosis'. The second meiotic division is
also divided into four stages.
i. Prophase II
ii. Metaphase II
iii. Anaphase II and
iv. Telophase II
PROPHASE - II
There is no relational coiling between sister chromatids. At the end, nucleus, nuclear envelope
disappears and spindle apparatus is organized.
Cytokinesis
Dyad divides into two parts. One parent cell produces from haploid daughter cells after meiosis.
The four daughter cells present together and are known as tetrad.
Significance of Meiosis
• Meiosis play an important role in all living organisms
• It helps in maintaining the chromosome number constant in a species.
• Meiosis results in production of gametes with haploid chromosome number.
• Union of female and male leads to formation of Zygote which receives ½ chromosome
number from female and ½ chromosome number from male, thus the original somatic
chromosome number is restored.
• Meiosis facilitates segregation of independent assortment of chromosome and genes.

26
 • Recombination of genes results in Creation of Variability which is essential for evolution
of new crop plants.
• In sexually reproduction crops Meiosis helps for continuity of generation.
zygote formation and embryo development
• The development of the embryo is called embryogenesis.
• In organisms that reproduce sexually, once a sperm fertilizes an egg cell, the resultant cell
is called as zygote that has half of the DNA of each of two parents.
• In plants, animals, and some protists, the zygote will begin to divide by mitosis to produce
a multicellular organism. The result of this process is an embryo.
• Plant embryogenesis is the process that produces a plant embryo from a fertilized ovule
by asymmetric cell division and the differentiation of undifferentiated cells into tissues and
organs.
• It occurs during seed development, when the single-celled zygote undergoes a
programmed pattern of cell division resulting in a mature embryo.
Identical and fraternal twins
• A twin is one of two offspring produced in the same pregnancy
• Identical twins (monozygotic) : they develop from one zygote that splits and forms two
embryos
• Fraternal twins (dizygotic ) - they develop from two separate eggs that are fertilized by
two separate sperm.

27
 Lecture 4. Chromosome structure, chemical composition, nucleosome, centromere,
telomere, euchromatin, heterochromatin, NOR, satellite chromosome, karyotype,
ideogram – chromosome banding.

CHROMOSOME
Chromosomes are rod shaped, dark stained bodies seen during metaphase. The term
'chromsome' was first used by Waldeyer in 1888. (Chrom- coloured soma =body), deeply stained,
while cytoplasm remained unstained. Each speceies has a definite chromosome number. Each
species has a definite chromosome number, represented by 2n. Somatic cells contain two copies
of each chromosome, which are identical in morphology, gene content and gene order and they are
known as homologous chromosomes. Gametic chromosome number is precisely one half of the
somatic number, is represented by 'n' zygote is produced by fusion of one male and one female
gamete (n+n=2n).
MOROPHOLOGY
Cell division, the following structural features can be seen under light microscope by
staining.
1. Chromatid
2. Centromere
3. Telomere
4. Secondary constriction and satelite
5. Chromosome
CHROMATID

28
 It is the structural and functional unit of chromosomes. At Metaphase, each chromosome
appears to the longitudinally divided into two identical parts, each of which is known a
'Chromotid'. The chromatids of a chromosome appear to joined the giller at a point called
'centromere'. The two chromatid making up a chromosome are produced through replication of a
single chromatid, they are referred as 'Sister chromatids'. In contrast the chromatrids of
homologous chromosomes are known as non-sister chromatids.

CENTROMERE
The region where thre two sister chromatids of a chromosome appeared to held together is
known as 'centomere' under light microscope, centomere generally appears as a constriction in the
chromosome, hre it is also termed as 'primary constriction'. Centromeres are the first part moving
towards the opposite poles during anaphase; the remaining regions lag behind and appear as if they
wree being pulled by the centomere. Therefore, chromosome movement is due to the centromeres
of chromosomes hence they are also known as 'Kinetochoes'. In most speceis each chromosome
has a single centromere in a fixed position which does not change except due to structural
chromosome aberrations. Therefore, the position of centromere serves as an important land mark
in the identification of different chromosomes of a species. Each chromosomes is divided into two
transverse parts by its centromere; these parts are called 'Arms'. On the basis of the position of
centromere, the chromosome may be divided into your classes.
i. Metacentric - Cntremere is at the centre of chromosome having equal arms and appeared as 'V'
shaped during anaphase.
ii. Submeta centric chromosoem - Centromere is on one side called 'Submedian. 'L' ' shaped
during anaphase.
iii. Acrocentric - When centromere is located close to one end, they are called as "Sub terminal 'j'
or rod shaped.
iv. Telo centric - Occasionally, the centromere appeared to be at one end of the chromosome,
called as 'Terminal' Rod shaped during anaphase. They are unstable. In most species each
chromosomes has a single centromere such chromosomes are termed as 'Monocentric'. But in some
speceis each chromosomes as 'Poly centric'- Polycentric chromosomes often break into smaller
chromosomal units each of which is stable and functions normally. Centromeres,

29
ccontain highly repetitive DNA called "Satellite -DNA" or "Sat- DNA", distinct from the rest of
the Chromosomal DNA. It constitutes about10% of total DNA present in the genome. In many
species Sat-DNA consists of only one sequence, while in others more thanone distinct sequences
are found.

TELOMERE
The two ends of chromosomes are known as 'Telomeres'. They are highly stable and do not
fuse with other chromosomes. It is generally accepted that, the structural integrity and individuality
of chromosomes is maintained due to the telomeres and that all stable chromosome ends are
composed of telomeres.

SECONDARY CONSTRICTION AND SATELITE

In some chromosomes a secondary constriction, in addition to that due to centromere
(primary constriction) is also present. It is known as "Secondary constriction. It is present in short
arm near one end, or in many chromosomes they are located in the long arm nearer to the
centromere. The region between the secondary constriction and the nearest telomere is known as
satellite. Therefore, chromosomes having secondary constriction are called "Satellite
Chromosome" or "Sat -Chromosomes. The position of secondary constriction in Sat-Chromsome
is fixed and remains constant. The number of Sat-Chromosomes in the genome varies from one
species to the other. The number of Sat. Chromosomes may ranged from 2,4,6 or 10, 13,14,15,21
and 22. Human somatic cells have 10 Sat Chromosomes. Nucleolus is always associated with the
secondary constriction of Sat. Chromosomes. Therefore secondary

30
constrictions are also called as "Nucleolus organizer Region" (NOR) and Sat-Chromosomes
are often referred as Nucleolus organism chromosome (NOC) NOR contains several hundred
copies of the gene coding for ribosomal RNA (r RNA).

CHROMOSOME
In some species (Maize, amphibian etc.,) chromosomes during Prophase I of meiosis,
particularly during pachytene stage, show small bead like structures called 'Chromomeres'. The
distribution of chromomeres in a chromosome is highly characteristic and constant, the patterns of
distribution being different for different chromosomes; homologous chromosomes show an
identical pattern.
• Flemming (1882) discovered two structures at cytological level
• Heitz (1928) coined as
• HETEROCHROMATIN - Found proximal to centromere, indistinguishable from
Euchromatin at metaphase- genetically inactive region
• EUCHROMATIN is genetically active and less contracted regions. Principal functional
regions
• Pachytene stage of meiosis is ideal for studying chromosome morphology
Two types of heterochromatin
• Constitutive heterochromatin or inherited
• Facultative heterochromatin or appearing during development

S.N Euchromatin Heterochromatin

1. Less condensed region More condensed region

2. Loosely coiled chromatin fibres More tightly folded chromatin fibres

3. Deeply stains- cell division; less stain Less stain - cell division; Deeply stain -interphase
-interphase

4. Genetically active - has unique DNA Genetically less active – has repetitive DNA

5. Can synthesise mRNA in vitro Unable to synthesise mRNA in vitro

31
 6. DNA replication is earlier – in the S phase DNA replication is at later stage –at the start of cell
division

7. This region is not sticky This region is sticky

8. Cross over frequency is more Cross over frequency is less

9. Does not show heteropycnosis show heteropycnosis

KARYOTYPE
The general morphology, i.e. the size of chromosomes, the position of centromeres, the
presence of secondary constriction and the size of satellite bodies of the somatic chromosomes
complement of an individual constitutes its "Karyotype". It is represented by arranging the
chromosomes in a descending order of size keeping their centromeres in a straight line. Each
chromosome in the karyotype is designated by a serial number according to its position. A perfectly
symmetrical karyotype has all metacentric chromosomes of the same size. Karyotypes showing a
deviation from this state are called asymmertical. It is believed that, perfectly symmetrical
karyotypes represent a primitive state from which more advanced asymmetrical Karyotypes have
evolved through structural changes in chromosomes.
Idiograms: Diagrammatic representation of chromosome arrangement based on size and length.
Idiograms of larch species: related species often have different karyotypes (idiograms) as is
shown by these idiograms of Larix decidua and L. sibirica.

32
Chromosome banding techniques
A chromosome banding pattern is comprised of alternating light and dark stripes, or bands, that
appear along its length after being stained with a dye. A unique banding pattern is used to identify
each chromosome and to diagnose chromosomal aberrations, including chromosome breakage,
loss, duplication or inverted segments.

Chromosome banding has since become a standard and indispensible tool for cytogenetic
analysis., and several banding techniques have been developed::
Q banding: chromosomes are stained with a fluorescent dye such as quinacrine . Chromosomes are
treated with quinicrine mustard solution, a fluorescent stain, to identify specific chromosomes and
structural rearrangements. It is especially useful for distinguishing the Y chromosome (also Y
bodies in interphase nuclei) and various polymorphisms involving satellites and centromeres of

33
specific chromosomes.
G banding: produced by staining with Giemsa after digesting the chromosomes with trypsin.
Chromosomes are G-banded to facilitate the identification of structural abnormalities. Slides are
dehydrated, treated with the enzyme trypsin, and then stained.
C banding: chromosomes are treated with acid and base, then stained with Giesma stain. C-
banding stains areas of heterochromatin, which is tightly packed and repetitive DNA. NOR-
staining, where NOR is an abbreviation for "nucleolar organizing region," refers to a silver
staining method that identifies genes for ribosomal RNA that were active in a previous cell cycle.
R-banding methods are useful for analyzing deletions or translocations that involve the telomeres
of chromosomes. R-banding is the reverse pattern of G bands so that G-positive bands are light
with R-banding methods, and vice versa. R-banding involves pretreating cells with a hot salt
solution that denatures DNA that is rich in adenine and thymine. The chromosomes are then
stained with Giemsa. R-banding is helpful for analyzing the structure of chromosome ends, since
these areas usually stain light with G-banding.
T-banding is used to stain the telomeric regions of chromosomes for cytogenetic analysis.
Telomeric (or terminal) banding was first reported by Dutrillaux, who used two types of controlled
thermal denaturation followed by staining with either Giemsa or acridine orange. The T bands
apparently represent a subset of the R bands because they are smaller that the corresponding R bands
and are more strictly telomeric..

Lecture 5. Types of chromosomes based on position of centromere, based on structure and

function: normal and special chromosomes - polytene, lampbrush, based on the role in sex
determination: autosomes and allosomes, Other types of chromosomes - B, ring and
isochromosomes.
Special chromosomes
1. Polytene chromosomes
34
• Also called as Giant chromosomes / Salivary gland chromosomes. First reported by Balbiani
in 1881
• The nuclei of the salivary gland cells of the larvae of Drosophila have unusually long and
wide chromosomes, 100 or 200 times in size of the normal chromosomes.
• The salivary gland cells do not divide after the glands are formed. But their chromosomes
replicate several times (a process called endomitosis) and become exceptionally giant – sized
to be called polytene chromosomes
• The polytene chromosomes of the salivary gland cells of D. melanogaster contain 1000 to
2000 chromosomes, which are formed by nine or ten consecutive multiplication cycles
and remain associated parallel to each other.
• polytene chromosomes have alternating dark and light bands along their length.
• The dark bands are comparable with the chromomeres of simple chromosomes and are disc-
shaped structures occupying the whole diameter of chromosome. They contain euchromatin.
• The light bands or inter bands are fibrillar and composed of heterochromatin.
• The swollen regions are known as chromosome “puffs” or Balbiani rings. They are the
regions of genetic activity .
• Chromosome puffs are diffuse uncoiled regions of the polytene chromosome that are
sites of RNA transcription. A Balbiani ring is a large chromosome puff.
• Such puffs change location as development proceeds, at specific locations.
• The presence of a specific puff is related with the appearance of a specific protein

35
 Polytene chromosome

2. Lampbrush chromosome One of the large chromosomes found in the eggs (primary
oocytes) of amphibians, with paired loops which extend from most of their chromomeres
giving a furry, brush-like appearance under the microscope. They are particularly obvious
at the diplotene stage of meiosis.Lampbrush chromosomes occur during the diplotene
stage of meiosis I. Lampbrush chromosomes are meiotic bivalents, each consisting of 2
sister chromatids. Each halve-bivalent is represented by two long strands that form many
brushlike loops along the main axis of the chromosome. The outgrowths make DNA
available for transcription during the maturation of the egg. Usually there is a little gene
expression at meiosis, so it is not so easy to identify the activities of individual genes. Giant
chromosomes in the lampbrush form can solve this problem, since they allow the
individual transcription units to be examined. Lampbrush and chromosomal puffs in a cell
indicate that the transcription of tRNA is taking place.

36
 Lampbrush chromosomes

3 B-Chromosomes
– Many plant (maize) and animal (insects and small mammals) species, besides having autosomes (A-
chromosomes) and sex-chromosomes posses a special category of chromosomes called B-
chromosomes without obvious genetic function. These B-chromosomes usually have a normal
structure, are somewhat smaller than the autosomes.

37
4. ISO Chromosomes- An isochromosome is a chromosome in which both arms are identical. It is
thought to arise when a centromere divides in the wrong plane, yielding two daughter chromosomes,
each of which carries the informations of one arm only but present twice. The isochromosomes are
formed during mitosis and meiosis.
If a gamete having a isochromosome is fertilized by a normal gamete, the zygote will possess an
unbalanced karyotype.
In Drosophila, the misdivision of centromere of telocentric X chromosome changes that into an
“attached-X” isochromosome, In man X-isochromosome causes the disease called gonadal dysgenesis.
An isochromosome is a chromosome that has lost one of its arms and replaced it with an
exact copy of the other arm

5. Ring chromosome
• A ring chromosome is a chromosome whose arms have fused together to form a ring.
• A ring chromosome is denoted by the symbol r.
• Ring chromosomes may form in cells following genetic damage by mutagens like
radiation, they may also arise spontaneously during development.
• Ring chromosomes found in prokaryotes
• Ring chromosomes otherwise called as genophores
• Ring chromosomes were Also reported in humans and drosophila

38
 • Ring chromosomes thoroughly studied in maize by Mc Clintock
• Ring chromosomes are meiotically unstable

Lecture 6. Chromosomal aberration: Variation in chromosome structure – deletion,

duplication, inversion and translocation – genetic and cytological implications.

Types of structural chromosomal aberrations

The chromosomal aberrations may remain confined to a single chromosome or may extend to
both of the member of the homologues pair and, therefore, may be of following types:
A. Intrachromosomal aberrations,
B. Interchromosomal aberrations.
A. Intrachromosomal Aberrations
When aberrations remain confined to a single chromosome of a homologous pair, they are called
intrachromosomal or homosomal aberrations.
1. Deficiencies (Deletions): Loss of a portion of segment from a chromosome is called Deletion
Two types - Terminal deficiency
Intercalary or interstitial deficiency.
Terminal deficiency
In deletion or deficiency type intrachormosomal aberration a chromosomal lacks either in an
interstitial or terminal chromosomal segment which may include only a single gene or part of a
gene.
Intercalary or interstitial deficiency: Loss of a portion of segment from a chromosome from the
intermediate portion or between telomere and centromere.
• Intercalary deletions are more common than terminal deficiency
• The deleted portion may have one / two / several genes

39
Genetic Significance of Deficiencies
Lethal effect: Organisms with homozygous deficiency usually do not survive to an adult stage
because a complete set of genes is lacking.

2. Duplications (Additions)
Duplication occurs when a segment of the chromosome is represented two or more times in a
chromosome of a homologous pair. This extra-chromosomal segment may be a free fragment with
a centromere or a chromosomal segment of the normal complement
• Reported by Bridges (1919) in Drosophila
• Recent reports is on several crops – rice , wheat, maize, Tobacco, Tradescantia, Barley

40
Genetic significance of Duplications
• The duplications of chromosomes are not deleterious to the organism like the deficiency,
but, they usually protect the organism from the effect of a deleterious recessive gene or
from an otherwise lethal deletion.
• some duplications are useful in the evolution of new genetic material. In an organism with
duplications, because the old genes can continue to provide for the present requirements of
the organism, the superfluous genes may be free to mutate to new forms without a loss in
immediate adaptability.
• Large duplications can reduce the fertility as a result of meiotic complication, and in this
way reduce their own probability of survival (Sybenga, 1972).
• Relocation of chromosomal material without altering its quantity may result in an altered
phenotype, this is called position effect.

INVERSIONS
An inversion is an intra-chromosomal aberration in which a segment is inverted 180 degrees. For
example if a chromosome has segments in the order of 1-2-3-4-5-6 and breaks occur in regions 2-
3 and 5-6 and the broken piece (3-4-5-) is reinserted in reverse order, then the inverted chromosome
will have segments in order of 1-2-5-4-3-6

41
Types of inversions
The inversions are of following types:
i) Pericentric inversions – When the inverted segment of chromosome includes or contains
centromere, then such inversions are called heterobranchial or pericentric inversions.
ii) Paracentric inversions – When the inverted segment includes no centromere and the
centromere remains located outside the segment, then such type of inversion is called
homobranchial or paracentric inversion.

42
Genetic significance of inversions
i) Simple inversions do not have primary phenotypic effects other then on chromosome shape.
Frequently, however, some DNA at a break point has been damaged and this may result in an
observable mutation, often recessive (e.g., clB lethal mutation in Drosophila).
ii) Due to inversion a peculiar kind of position effect occurs. The position effect is caused by the
transfer of a gene from a euchromatic segment to the vicinity of heterochromatic segment.
Heterochromatinization may then extend into a displaced, originally euchromatic region and
suppress the transcription of the gene in it.
iii) Normal linear pairing is not possible in inversion heterozygotes. The difficulties encountered
with pairing cause a reduction of exchange (crossing over) in and around the inversion.
iv) They maintain heterozygosity from generations to generations.
B. Interchromosomal aberrations

43
When breaks occur in non-homologous chromosomes and resulting fragments are interchanged by
both of the non-homologous chromosomes, the inter-chromosomal or heterosomal aberrations
occur. The inter-chromosomal aberration is of following type:
TRANSLOCATION It is an inter-chromosomal aberration where in exchange of chromosomal
segments occurs between non-homologous chromosomes
RECIPROCAL TRANSLOCATION: Translocation involves the shifting of a part of one
chromosome to another non-homologous chromosome. If two non-homologous chromosomes
exchange parts, which need not be of the same size, the result is a reciprocal translocation. The
reciprocal translocation may be of following types:
Homozygotic translocation - In homozygotic translocation normal meiosis occur and cannot be
detected cytologically. Genetically they are marked by altered linkage group by the fact that a
gene with new neighbors may produce a somewhat different effect in its new location (position
effect).
Heterozygotic translocation – In heterozygotic translocation a considerable degree of meiotic
irregularity occur. During meiosis, an individual which is heterozygous for a reciprocal
translocation must form a cross-shaped configuration in order to affect pairing of all
homologous segments. This cross-shaped configuration often opens out into a ring as
chiasmata terminalize. The meiotic products (gametes) are of three types –normal balanced
and unbalanced gametes
Genetic significance of Heterozygotic Translocation:
1. The heterozygous translocation produce semi-sterile organisms because between half and two
third gametes fail to receive the full complements of genes required for normal development
of sex.
2. Some genes which formerly assorted independently, exhibit linkage relationships after
translocation has occurred; a single reciprocal translocation will reduce the number of linkage
groups by one.
3. The phenotypic expression of a gene may be modified when it is translocated to a new position
in the genome (position effect).

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 Lecture7. Chromosomal aberration: Variation in chromosome number – euploid,
aneuploid, types of aneuploids and their origin; Nondisjunction - Klinefelter syndrome
and Turner syndrome; Definition of eugenics and euthenics

The term genome refers to a complete set of chromosomes of a diploid species. A deviation
from the diploid state represents a numerical chromosomal aberration which is often referred to as
heteroploidy. Individuals possessing variant chromosome number are known as heteroploids.

Numerical changes in chromosomes

• A) Alterations in whole chromosome sets (Euploidy)
• B)Additions or subtractions of individual chromosomes (Aneuploidy)
Euploidy individuals having the chromosome number which is an exact multiples of the basic or
genomic number
Aneuploidy is the term for cells, tissues, and individuals with excess or lacking one or few
individual chromosomes; It is a change in the number of chromosomes that can lead to a
chromosomal disorder
− hyperploidy is a type of aneuploidy when there is an excess number of
chromsomes (trisomics, tetrasomics)
− hypoploidy is another type of aneuploidy when one or more number of
chromosomes are lacking(monosomics, nullisomics).
Types of Aneuploid

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Nullisomics(2n – 2): individuals from which one chromosome pair is missing
Monosomics(2n – 1): those lacking a single chromosome
Double monosomic(2n – 1 – 1) : individual has two chromosomes missing, but the two
chromosomes belong to two different chromosome pairs.
Trisomic(2n + 1): An individual having one extra chromosome
Double trisomic (2n + 1 +1): that having two extra chromosomes each belonging to a different
chromosome pair
Tetrasomic (2n + 2): When an individual has an extra pair of chromosomes
Origin of Aneuploids:
• Non-disjunction – failure of separation of paired chromosomes

• occurs when paired chromosomes do not separate either during meiosis I or meiosis II.

Sources of Aneuploids

Aneuploids in human disease

Monosomy (2n-1) condition

Turner’s Syndrome females have lost one of the X chromosomes (XO), sterile

Trisomics (2n+1) condition

Klinefelter’s Syndrome: XXY feminized males; sterile

Down Syndrome: Result of a trisomy of chromosome #21 (although a few cases due to a
translocation )
46
Trisomy 13 (Patau syndrome) and trisomy 18 (Edwards syndrome) can also survive to birth.

Eugenics" The study of the agencies under social control that may improve or impair the
racial qualities of future generations either physically or mentally.

Euthenics: the study of methods of improving human well-being and efficient functioning by
improving environmental conditions. (Or) Measures to improve the environment in order to
improve health, appearance, behavior, or well-being of society.

Euphenics: Measures to improve the individual or phenotype (the body) by biological or

medical means.

A summary of terms used o describe heteroploidy (variation in chromosome number):

Term Type of change Symbol*

47
48
Euploids
In euploids, the chromosome number is an exact multiple of the basic or genomic number.
Euploidy is more commonly known as polyploidy. When all the genomes present in a polyploidy
species are identical, it is known as autopolyploid and the situation in termed as autopolyploidy.
In the case of allopolyploids, two or more distinct genomes are present. Euploids may have 3
(tripoid), 4 (tetraploid), 5 (pentaploid), 6 (hexaploid), 7 (heptaploid), 8 (octaploid) or more
genomes making up their somatic chromosome number.
In case of autopolyploidy, they are known as autotriploid autotertaploid, autopentaploid,
autohexaploid, autoheptaploid, autooctaploid and so on, while in the case of allopolyploidy they
are termed as allotriploid, allotetraploid, allopentaploid, allohexaploid, alloheptaploid,
allooctaploid etc.
Amphidiploid is an allopolyploid that has two copies of each genome present in it and, as a
consequence, behaves as a diploid during meiosis. A segmental allopolyploid contains two or more
genomes, which are identical with each other, except for some minor differences.
Euploidy includes monoploids, diploids and polyploids.
Monoploids: Monoploids contain a single chromosome set and are characteristically sterile.
In other words monoploids have the basic chromosome number (x) of a spec ie. Monoploids

49
 (x) differ from haploids (n) which carry half or gametic chromosome number. In a true
diploid species, both monoploid and haploid chromosome number are same (i. e. x=n).
Haploid: Haploid is a general term used to designate the individuals or tissues with a
gametic chromosome number i.e. n.

Differences between monoploids and haploids

Monoploids Haploids
Represent gametic chromosome number of a Represent gametic chromosome number of any
diploid species species
Denoted by ‘x’ Denoted by ‘n
Monoploids are always haploids Haploids cannot always be monoploids
Contain single set of genome May contain one or more copies of genome.

50
 Lecture 8. Polyploid - auto and allopolyploids, their characters; meaning of genome;
evolution of wheat, Triticale, cotton, tobacco, Brassicas,

POLY PLOIDS

Individuals with more than two sets of chromosomes are called ‘ polyploid’. Poly ploid in
otherwise called as ‘Euploidy’.
Individual with one set of chromosomes – haploid or Monoploids.
Individual with two sets of chromosomes – True diploid
Most plants and animals posses two sets of chromosomes.
GENOME
The complete set of chromosome found in the gamete of a true diploid is called a
genome. e.g. p. glaucm, 14 chromosome 2 sets n=7, 2n=14.
POLYPLOIDS
Individuals with more than two sets of chromosomes are called ‘ polyploid’..
Somatic No. (2n) Multiples of Basic No.(x-12) Level of ploidy
24 2x Diploid
36 3x Triploid
48 4x Tetraploid
60 5x Pentaploid
72 6x Hexaploid

CLASSIFICATION
POLYPLOIDS

Based on origin Based on genome

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 Natural Induced Autopolyploids Allopolyploids

Autopolyploid: when same set of chromosomes of a genome are increased in number, it is

autopolyploid. For example, if a diploid species has two similar sets of chromosomes / genomes
designated as AA, an autotriploid will have three similar (AAA) genomes and autotetraploid will
have four similar (AAAA) genomes

Origin of autopolyploids

− The effective method to obtain autopolyploids is using colchicine.

− Colchicine is a spindle fiber poison or suppressant.
− It inhibits the spindle mechanism at mitosis, resulting multiples of normal
chromosome number.

Types of Autopolyploids

Auto triploids (3x): Auto triploids have three complete sets of genomes of the same species in
somatic cell. AAA

• Autotriploids can be produced by crossing diploids with their corresponding

autotetraploids (2x x 4x).
• The high sterility of autotriploids has been explored in plant breeding.
• Triploid bananas (2n = 33) are vigorous but seedless and therefore preferred for food
consumption.
• Triploid watermelons have only undeveloped seeds.
• Triploid is also applied in seedless Citrus cultivars

Important triploid plants include, some potatoes, bananas, watermelons and Winesap apples

Autotetraploids: four copies of the genome of same species (AAAA or BBBB) are present.
• Examples of autotetraploids are alfalfa, coffee, peanuts and McIntosh apples.
• These also are larger and grow more vigorously
• All of these crops must be propagated asexually

Use of Autopolyploids in Plant breeding

• i) Autopolyploid manifests greater vegetative growth but reduced seed production.
• This implies that autopolyploid induction would be more useful for vegetative parts of the
plants, such as forage, or root, but not the seed.
• ii)Autopolyploid produced from diploids with lower chromosome number have been
relatively more successful.
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Allopolyploids A polyploid containing genetically different chromosome sets from two or more
species is known as allopolyploid. (AABB)
• Allopolyploid are polyploids with chromosomes derived from different species.

Amphidiploid It is an allopolyploid (allotetraploid) which arises by combining genomes of two

different species. The amphidiploids are fertile due to the presence of homologous chromosomes
and behave as a diploid during meiosis.

Role of polyploidy in evolution of crops - wheat, cotton, Tobacco and Brassica.

Natural allopolyploids: Inter-specific crossing followed by chromosome doubling in nature

have resulted in origin of some natural allopolyploid crops like cotton, tobacco, mustard,
wheat, etc. The origin of some natural allopolyploid crops is briefly presented below:

(i)Wheat: The common or bread wheat, Triticum aestivum is an allohexaploid. It has two
copies each of the genomes A, B and D and its somatic complement is represented as AA BB
DD. The sources of A and D genomes are more or less unanimously accepted as Triticum
monococcum (AA) and Triticum tauschii (DD) (formerly Aegilops squarrosa –goat grass),
respectively. There is considerable doubt about the source of B genome. According to one
hypothesis, Aegilops speltoides may be the source of this genome.

(i) Cotton: The new world cotton (Gossypium hirsutum) is an interesting example of
allopolyploidy.

53
(ii)Tobacco: There are two cultivated species of tobacco. i. e. Nicotiana tabacum and
Nicotiana rustica.

a) Nicotiana tabacum is an allotetraploid and available evidence suggests that it is derived

from a cross between Nicotiana sylvestris x Nicotiana tomentosa

b) Nicotiana rustica is believed to be an amphidiploid obtained from a cross between

Nicotiana paniculata and Nicotiana undulata

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(ii)Brassica:

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Lecture 9. Pre-Mendelian ideas about heredity – Vapour and fluid theory, Magnetic power
theory, Preformation theory, Lamarck’s theory, Darwin’s theory, Germplasm theory and
Mutation theory.
Early concept:
Among the biological sciences the science of genetics originated 1900 with the rediscovery
of Mendelian Principles. Though pre-historic plants and animal breeders employed hybridizations
and selection they were not aware of principles of genetics.
Early works forwarded various speculation and theories explain the phenomenon of
heredity. The ideas of early workers can be grouped into the following headings.
1. Vapour and fluid theory:
Early Greek Philosophers thought hereditary information of the parents exist in the form
of vapour and fluid. Pythagorus 500 B.C. proposed that the moist vapour from dry nurves and
other parts of the body organs from the nerves form an embryo in the uterus of the female.

56
2. Magnetic power (Harvey (1578 - 1657)
He proposed the uterus had some magnetic power to consume an embryo.
3. Pre-formation theory

With discovery of Sperms and egg by 17 th century biologist pre-formation theory was
proposed. According to this new individuals of completely pre-formed in miniature size in
gametes.
4. Epigenetic theory ( Wolf) 1738
He proposed that each egg had a granule which gradually develops into various organs of
the embryo.
5. Particulate - Maupertius (1698 - 1759)
Maupertius proposed that both the parents produce the gametes. Egg and sperm united to
produce embryo. Each organ of embryo is made of two parts of embryo one is come from male
() another one female ().
6. Theory of Lamarck (1744 - 1829)
Lamarck proposed that environmental changes cause modification in organism and that
such modification transmitted to subsequent generation. That is the character acquired in one
generation to another generation. Acquire characters are inheritable.

e.g. Giraffie live in the interior part of the Africa. So it feed on the leaves of tall tree and
to strain itself continuously to reach them. Such exercise caused the legs and necks to grow in
length. The increased length was inherited by the progeny touched the legs and neck and over
generation it will continue. This has evolved the present day 6 metre giraffie.

7. Theory of pangenesis - Charles Darwin (1809 - 1882)

Darwin proposed theory of natural selection. Many individuals of each species and there
is always existence. If hereditary differences occurs with in a species of plants natural selection
allowed only the fittest individuals of the species to survive and eliminating others. This is known
as survival of fittest due to natural selection.
In 1868 Darwin published origin of species. He also proposed theory of Pangenesis . he
called it hereditary particles. Germules that are produced every part of the body during the life

57
time. The germules from all the organs are transported to the gonads. The gonads distributed
gametes if male () gametes unit during fertilisation, germmules from the migrate in respective
organ and determine the development of organ. Organ is modified some way. The gemmules of
the organ also modified accordingly which is transmitted to the offspring through gametes.

Germplasm theory(Weismann) (1814 - 1914)

Weismanns suggested the reduction in chromosome number takes places during formation
of eggs / sperm and the original number resorted when eggs sperm fused. According to him
acquired character not inherited. To prove this in mice to cut off tails 22 generation at last 1592.
All of them having normal fail end. If the acquired characters are inherited the tails condition of
mice produced by cuttings of their tails should be transmitted to offsprings. If the theory of
pangenesis was correct the germmules for tail would be absent in the gamets of the tail less mice
and their progency would be tails less.
Weismann proposed germplasm theory of heredity particles called IDS (Genes) situated on
idants (chromosome) constitute the germplasm. The germplasm transferred to parents to offsprings
and give rise body or some. The germplasm is independent of the body. Whatever happens to the
body has not effect on the germplasm. This was prooved by ovary transplantation in guinea pigs.
When albino guinea pigs are mated with alblono only albino progenies are produced.
Castle & Phillips (1909) removed the ovary of the albino guinea pigs and grafted in their
place the ovary of guinea pig. The albino animal with the ovary of the black one when mated with
the albino on the offsprings is black only. This proved the germplasm is not affected by body.
The history of genetics can therefore be reviewed under three periods (1) Pre-Mendelian,
(2) Mendelian, and (3) Post-Mendelian.

PRE-MENDELIAN PERIOD
Some of the scientists prior to Mendel tried to account for the differences existing between
individuals and suggested theories to explain their inheritance. The most important theories are the
following:

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Theory of Lamarck
The French biologist Jean Baptiste de Lamarck (1744-1829) proposed the theory that
environmental changes cause modifications in organisms and that such modifications are
transmitted to subsequent generations. He believed that environment acts directly on plants and
indirectly on higher animals.
Lamarck said that changes in environmental conditions create new needs in animals.
Conscious efforts of the animals to adapt to the environment involves the use of certain organs,
thereby causing them to become large, strong and well-developed. Other organs are not used and
so become smaller, weaker and less well-developed. Such bodily changes are called acquired
characters since an animal achieves them by its own exertions to adapt to the environment.
Acquired characters, according to Lamarck, are then passed on to the offspring of the organism
that acquired them, and new species originate by accumulation of these modifications.
The giraffe dwells in the interior arid parts of Africa where there is not much herbage.
According to Lamarck, the giraffe was obliged to feed on the leaves of tall trees and to strain itself
continuously to reach them. Such exercise caused the necks and legs to grow in length. The
increased length was inherited by the progeny, which, in turn, stretched their necks and legs and
transmitted their increased length to their own offspring. Thus has evolved the present day six-
metre high giraffe.
Detailed studies have failed to show that acquired characters are inherited. Most biologists
have therefore abandoned the theory of inheritance of acquired characters, otherwise known as
Lamarckism.
Darwin's Theory
In 1858, Charles Darwin (1809-1881) and Wallace independently proposed the 'Theory of
Natural Selection'. According to this theory, many more individuals of each species are born than
can possibly survive and consequently there is always a struggle for existence. If hereditary
differences occur within the wild species of plants, nature will eliminate some and select others.
Over-production, struggle for existence, hereditary variations and survival of the fittest are
thus the important principles of the theory of natural selection.

59
 Ten years after the publication of the Origin of Species (1859), Darwin adopted the
doctrine of the inheritance of acquired characters but he proposed a new theory of how it
happened. He modified the views of Spencer and proposed the 'Hypothesis of Pangenesis' (1868).
Darwin assumed that hereditary particles termed pangenes of gemmules, are produced by
every part of the body during the life time of an organism and that, these assume the characters of
the various parts of the body from which they were derived, together with whatever modifications
the latter may have acquired. Eventually all the pangenes accumulate to form the germ cells which
give rise to the new individual, thus ensuring the development of the parental characters and
inheritance of acquired characters.
Weismann's Germplasm Theory
Weismann(1834-1914), a German zoologist, suggested in 1887 that a reduction in
chromosome number took place during the formation of the egg and the sperm, and that the original
number was restored when the egg and the sperm fused. In 1892, he suggested that the maternal
and paternal chromosomes separated during the reduction division and that they recombined when
the gametes united.
According to Weismann's Germplasm Theroy of Heredity', the hereditary particles called
ids (what we now call as genes) situated on idants (what we now call chromosomes) constituted
the germplasm. The germplasm is handed down from parent to offspring and it gives rise to the
body or soma (somatoplasm) whose character it determines. The germplasm is independent of the
body and whatever happens to this body has no effect on the germplasm which is contained within
it.
According to Weismann, acquired characters cannot therefore be inherited. To prove this
he cut off the tails of mice for twenty-two generations and found that the progeny consisting of
1,592 individuals had tails of normal length.
The independence of the germplam from the somatoplasm was shown by the ovary
transplantation experiment in guinea pig. Ordinarily, when albino guinea pigs are mated with
albinos, only albinos are produced. Castle and Phillips removed the ovaries of an albino guinea
pig and grafted in their place the ovaries of a black guinea pig. The albino animal with the ovary
of the black one was then mated with an albino. All the offspring were found to be black, thereby

60
proving that the germplasm (i.e., the ovary from the black guinea pig) is not affected by the
somatoplasm (i.e., the body of the albino).

De Vries' Mutation Theory

Charles Darwin believed that evolution is due to natural selection of small hereditary
variations occurring among individuals of any species. Bateson did not agree with Charles Darwin.
He believed that evolution is due to large discontinuous variations. De Vries (1848-1935)
introduced the term `mutation' for these large, discontinous changes in the genotype and proposed
the `Mutation Theory', according to which sudden hereditary changes lead to evolution.
De Vries (1901) observed that the evening primrose Oenothera lamarckiana, a native of
America, was growing wild in Holland. In a population of this weed, he observed some plants
which differed in some characters from the typical Oenothera lamarckiana. Since it is a self
-fertilised species, he felt that these variants have arisen suddenly rather than as hybrids. He
transplanted them to his garden and studied them for several years. He observed that variations
continued to arise spontaneously and that these variations were inherited. He called these drastic
changed as mutations and maintained that mutations play an important role in the evolution of new
species.

Lecture 10. Work of Mendel – Characters studied reasons for Mendel’s success, Law of
dominance, Law of segregation and Law of independent assortment. Rediscovery of
Mendel’s work

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Work of Mendel
Gregor John Mendel was born in 1822 near Brunn in Austria. In 1843, Mendel entered
Augustinian monastery at Brunn. He completed his theological studies in 1848 and was appointed
as a substitute teacher in high school. In 1851 – 53, the monastery sent him for studies at university
of Vienna. He continued as a substitute teacher for 12 more years. In addition he worked as a priest
in the local church. He lived in a house located within the premises of the church. He began to
collect pea seeds for his experiments in 1857 from commercial seed growers all over the Europe.
He conducted all his experiments within the kitchen garden of his house with the help of his own
resources.

Mendel carried out his experiment in the monastery gardens from 1856 – 63. His
experimental material is garden pea (Pisum sativum). He studied seven contrasting characters.
He presented the result of his experiment before “The National History society at Brunn”. His
paper entitled "Experiments in plant hybridization" was presented in German language. No.
one appreciated the importance of his work until 1900. Gregor Johan Mendel died in 1884, at the
age of 62 years.
Seven Characters studied by Mendel
Character Dominant Recessive
1. Seed shape Round Wrinkled
2. Petal colour Purple White
3. Cotyledon colour Yellow Green
4. Pod colour Green Yellow
5. Pod shape Full Constricted
6. Position of flower Arial Terminal
7. Length of stem Tall Dwarf

PEA as an experimental material

Pea offered several advantages as an experiment material.

62
i. In the pea varieties available commercially, several characters had two contrasting form which
were easily distinguishable from each other.
ii. The flower structure of pear ensured self pollination this was experimentally verified by
Mendel. This greatly facilitated the production of F2 and F3 progeny as well as avoided
contamination by foreign pollen.
iii. Pea flowers are relatively large. Therefore emasculation and pollination is quite easy, which
allows easy artificial hybridization in pea.
iv. The duration of pea crop is of a single season. As a result, every year one
generation of pea can be grown.
v. Pea seeds are large and present no problem in germination. Pea plants are relatively easy to
grow and each plant occupies only a small space. This persists a
large number of plants to be grown in a relatively small area. (In addition, Mendel worked in Raj
mash, P. vulgaris)
Mendel’s findings:
1. Both male and female make equal contribution to the development of character in progeny,
since the results from reciprocal crosses are identical.

2. In F1 generation, character of only one of the parent is expressed. This is known as

dominant character. The character of the other parent which is not expressed is referred as
recessive.
3. In F2 characters of both the parents ie., dominant and recessive appeared in a definite

proportion of 3:1
4. The recessive character is not modified in F1 generation and its expression is prevented.

5. The dominant character may be of 2 types.

i) It may be pure like that in the parent producing progeny with the dominant trait.
ii) It may be a hybrid similar to F1 hybrid (heterozygote) producing ¾ progeny

with dominant character and remaining ¼ with recessive character.

Reasons for Mendel success:
1. He accurately diagnosed the weakness of earlier experimental materials, techniques and
approaches. He carefully avoided mistakes in his experiment.

63
 2. Mendel studied the inheritance of only one pair of contrasting character at a time.
3. He selected pea varieties, which have clearly different forms of one or more character.
4. He classified all the plants of a population on the basis of contrasting character under
studying and kept accurate record of the no. of plants in each category for every generation.
5. He carried out his experiments with great care, i.e he grow the parents in two seasons to
avoid mechanical mixture.
6. His knowledge on mathematics was a definite asset for the interpretation of his result.
The Laws of Mendel:
1. Law of dominance:
On crossing homozygous organisms for a single pair of contrasting characters, only one
character make its appearance in F1 generation and is named as dominant character.

2. Law of segregation (law of purity of gametes): (Mendel's First Law)

If two alleles are brought into a hybrid, they do not contaminate or blend with each other
but segregate and pass into different gametes.

3. Law of independent Assortment: (Mendel's Second Law)

When two or more pairs of alleles are brought into a hybrid, the segregation of any one
pair of allele is independent to that of segregation of any other pair of allele.

Rediscovery of Mendel’s work:

In the year 1900, Mendel’s paper was rediscovered. Three scientists working
independently of each other, de Vries in Holland, Correns in Germany and Tschermak in Austria,
arrived at the same conditions as those of Mendel. After this rediscovery, there was a spurt of
interest in the Mendel's findings and the science of Genetics was truly borne.

Mono hybrid

The progeny derived by crossing two individuals (or) strains which differ for one gene.

Di hybrid:

The progeny from a cross between two homozygous parent different for two gene.

Tri hybrid

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 The progeny from a cross between parents differing in three genes.
No. of Phenotypic ratio No. of No. of Genotypic ratio
gene phenotypes genotypes
1 (3:1) 2 3 (1:2:1)
2 (9:3:3:1) 4 9 (1:2:1:2:4:2:1:2:1)
3 27:9:9:9:3:3:3:1 8 27
Different kinds of 2n 3n
gametes produced

Where’ n’ is no. of genes for a character

Mendel's monohybrid cross
As mentioned earlier, Mendel studied the inheritance of each of the seven pairs of
contrasting characters selected by him in peas in separate crosses. For example, he crossed a variety
of peas having round seeds with a variety having wrinkled seeds. The hybrid seeds from this cross
were all round. He planted these hybrid round seeds and obtained an F2 generation consisting of

round and wrinkled seeds in the proportion of 3:1. Similarly, he crossed a variety of peas having
yellow cotyledons (the colour of which could be seen through the `transparent' or thin seed coats)
with a variety having green cotyledons. The hybrid seeds resulting from this cross were all yellow.
He planted these hybrid yellow seeds and obtained an F2 generation consisting of yellow and green

seeds in the proportion of 3 yellow : 1 green.

Mendel's dihybrid cross
Mendel then crossed a variety with round and yellow seeds with a variety with wrinkled
and green seeds. The F1 hybrid seeds were all round and yellow. The plants raised there from

yielded seeds of four sorts which frequently presented themselves in one pod. In all, 556 seeds
were yielded by 15 plants and of these, there were:
315 Round and yellow
108 Round and green
101 Wrinkled and yellow
32 Wrinkled and green

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 Mendel observed that the segregation for form of seed and colour of cotyledons
separately was in the ratio of 3 dominants: 1 recessive.

Round seeds (315 + 108) / 556 76.08%

=
Wrinkled seeds (101 + 32) / 556 23.92%
=
Yellow seeds (315 + 101) / 556 74.82%
=
Green seeds (108 + 32) / 556 25.18%
=

F2 ratio of hybrid F2 ratio of hybrid F2 ratio of hybrid round

round yellow and yellow

3Y = 9 RY (round yellow)
3R 1y = 3 Ry (round green)
3Y = 3 rY (wrinkled yellow)
1r 1y = 1 ry (wrinkled green)

Mendel concluded that the F2 of a cross involving two pairs of contrasting characters (i.e., a

dihybrid cross) shows four kinds of individuals in the ratio of 9 : 3 : 3 : 1.

Mendel's Second Law
Mendel arrived at the following conclusion:
When an individual forms gametes, the members of a pair of alleles always segregate (i.e.,
separate) from each other but the members of different pairs of alleles assort independent of each
other.
Mendel's Second Law of Inheritance or the Law of Independent Assortment can, therefore,
be expressed as follows:
The segregation in one pair of alleles is independent of the segregation in any other pair of
alleles.

Test cross- Crossing of F1 with recessive parent

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That the dihybrid forms four kinds of female gametes and four kinds of male gametes in equal
numbers can be shown by crossing it with the double recessive.
From a cross between the dihybrid as female and the wrinkled green seeded plant as male,
Mendel obtained 31 round yellow, 26 round green, 27 wrinkled yellow and 26 wrinkled green
seeds. As the recessive plant produces only one kind of male gamete, ry, a 1 : 1 : 1 : 1 ratio is
possible only if the dihybrid produces four kinds of female gametes, RY, Ry, rY and ry, in equal
numbers.

Back cross
Crossing of F1 with any one of the parent
From a back cross between the double recessive as the female and the dihybrid as the male, he
obtained 24 round yellow, 25 round green, 22 wrinkled yellow and 27 wrinkled green seeds. The
progeny is in the ratio of 1 round yellow : 1 round green : 1 wrinkled yellow : 1 wrinkled green,
thereby showing that the dihybrid produces four types of gametes, RY, Ry, rY and ry in equal
numbers.

Trihybrid ratio
Trihybrid is a hybrid resulting from a cross between parents differing in three genes.
As an example, we can consider Mendel's cross between pea plant with round seeds, yellow
cotyledons and grey-brown seed coats and one with wrinkled seeds, green cotylendons and white
seed coats. All the hybrid seeds resulting from this cross are round, yellow and grey-brown.
An individual heterozygous for three independently assorting pairs of alleles produces
eight types of gametes in equal numbers as follows:

RYB RYB
Ryb Ryb
RyB RyB
Ryb Ryb

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 Eight kinds of male gametes fertilising at random eight corrresponding kinds of female
gametes produce an F2 consisting of 64 possible combinations composed of 27 different

genotypes. Since R is dominant over r, Y over y, and B over b, the 27 different genotypes fall into
eight visibly different types (i.e., phenotypes) as follows:

3 B = 27 RYB (round, yellow, brown)

3Y 1 b = 9 Ryb (round, yellow, white)
3R 3 B = 9 RyB (round, green, brown)
1y 1 b = 3 Ryb (round, green, white)
3 B = 9 rYB (wrinkled, yellow, brown)
3Y 1 b = 3 rYb (wrinkled, yellow, white)
1r 3 B = 3 ryB (wrinkled, green, brown)
1y 1 b = 1 ryb (wrinkled, green, white)

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 Lecture 11. Chromosomal theory of inheritance. Allelic interactions – Dominance vs.
recessive, complete dominance, codominance, incomplete dominance, over dominance.

Chromosomal theory of inheritance: (Sutton and Boveri)

The idea of chromosomal basis of segregation and independent assortment was put forth
by Sutton in 1902. He explained the law of segregation and independent assortment on the behavior
of chromosomes during meiosis. The hypothesis that genes are located in chromosome was
designated by Sutton and Bovery. Sutton and Bovery hypothesis was known as chromosome
theory of inheritance. Evidence for chromosome theory of inheritance

1. Each somatic cell contains 2 copies of each gene i.e., A, a similarly each somatic cell
has 2 copies of homologous of each chromosome. Each somatic cell produced during
embryonic and subsequent development receives 2 copies of each gene present in the
zygote.

2. A gamete contains only one copy of a gene or one allele of a gene. This phenomenon is
termed as segregation. It is assumed that 2 alleles of a gene are located in the 2
chromosomes of a homologous pair. Separation of 2 homologous chromosomes during
anaphase I of meiosis will account for segregation of 2 alleles.

3. Several subsequent studies and Mendel's studies found that 2 or more genes assorted
independently to yield typical dihybrid, trihybrid ratios. Members of homologous
chromosomes assort independently to that of the other.

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 4. In 1901 Meclung discovered the accessory or x chromosome of grass hopper and stated
that, this chromosome was involved in sex determinanation since a specific chromosome
is involved in sex determination, the genes governing this trait may be located on x
chromosomes.

5. In 1910, T.H. Morgan presented a more direct evidence supporting the chromosome
theory of heredity. He found that the pattern of transmission of white eye gene was identical
with that of the x chromosome of Drosophila. This prompted Morgan to postulate that the
gene for white eye was located in the x chromosome.

6. In 1910, Morgan described the phenomenon of linkage and crossing over between 2 sex
linked genes in drosophila. He proved that the linkage between any 2 genes depends on the
distance between them in the chromosome. Morgan explained that the recombination of
linked gene is due to exchange of genetic material between homologous chromosomes.

7. When a segment of chromosome is inverted, the inverted chromosome shows as inverted

gene sequence corresponding to the inverted segment. Hence genes must be located in
chromosome.

8. In addition to structural changes in chromosomes, numerical chromosome changes also

furnish in favour of this theory. In monosomics and trisomics there will be significant
deviation from the normal inheritance pathern ie from 3:1 ration in F2 for a single gene.

Parallelism of the chromosome and genes.

i. Chromosomes occur in pairs one received from male () parent other received from female
() parent. Gene also exist in pairs (allele) one from  another .
ii. Each pair of chromosome differ from other pairs. Likewise each gene has individuality and
produce specific effect.
iii. At meiosis the member of one pair of chromosomes separate and go into different gametes.
According to law of segregation each gene separate from its allele and each one enter into
different gamete.
iv. The segregation of one pair of chromosome has been observed is independent of the
segregation in another pair chromosome. According to law of independent assortment,
segregation of one pair of allele to another pair of alleles is independent.
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Allelic interaction –interaction within the allele of same gene is called as intra-allelic
interaction or dominance
Types of dominance
1. Complete dominance:
In case of complete dominance, the phenotype produced by heterozygotes is identical with
that produced by homozygotes for the concerned dominant allele. The dominant allele in such a
situation is known is completely or fully dominant.
Eg: In peas, round seed shape is produced by the dominant allele W, while wrinkled shape
is determined by its recessive allele w. Seeds having the genotype Ww are round and
indistinguishable from those having the genotype WW. As a result, characters showing complete
dominance yield the typical 3:1 monohybrid ratio in F2.
2. Incomplete Dominance:
The phenotypic expression of heterozygote for a gene being intermediate between those of the two
concerned homozygote. Such a situation is known as incomplete or partial dominance.
Eg:- Flower colour in 40 O Clock plant
(Mirabilis jalapa)
RR x rr
(Red) (White)
F1 : Rr (pink)
F2 : 1 RR : 2 rr : 1 rr
Red : pink : white
Here the phenotypic ratio in F2 is 1: 2 :1 instead of 3:1 in Complete dominance
3. Co – Dominance: Both the alleles of a gene express themselves in the heterozygotes. As a
result, heterozysotes for the such genes posses the phenotype produced by both the alleles of such
genes.
Eg. Blood group antigens of man.
I A IB x I A IB
(AB) (AB)
I A IB : 2 I A IB : 1 IB IB
1 A: 2 AB : 1B
coat colour in cattle
CR CR x CW CW

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 (Red) (White)

F1 CR Cw (Roan)

F2 1 CR CR : 2 CR CW: 1 CW CW
Also called as mosaic dominance

4) Over dominance:
In case of some genes, the intensity of character governed by them is greater in
heterozygotes than in the two concerned homozygotes. This situation is known as over dominance.
Eg: - AA x aa
(120 cm) (100mm)
F1 Aa (140 cm)
F1 is superior to the dominant parent. It is called as Hetero, Super (or) over
dominance.

Lecture 13. Deviation from Mendelian inheritance – Non allelic interaction without
modification in Mendelian ratio – Batson and Punnet’s experiment on fowl comb shape.
Non allelic interaction with modification in Mendelian ratio – i.) Dominant epistasis
(12:3:1)
Gene interaction: For the determination of single phenotypic character, two alleles of a single
gene interacted in various way.
Eg: complete dominance, incomplete dominance or codominance. These kind of genetic
interactions occur in between the two alleles of a single gene is referred as Allelic interaction or
intra geneic interaction.
Non – allelic interaction or intergenetic interactism (or) Epistasis

Deviation from Mendelian inheritance – Non allelic interaction without modification in

Mendelian ratio – Batson and Punnet’s experiment on fowl comb shape

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 A classical case of two pairs of alleles affecting the same characteristic and producing in
the F four different phenotypes in the ratio of 9 : 3 : 3 : 1 was discovered in fowls by Bateson
2

and Punnett.
Each breed of pultry possesses a characteristic type of comb. The Wyandotte breed has a
comb known as the ‘rose’ comb, the Brahma has a ‘pea’ comb, the Leghorn has a ‘single’ comb
and the Malay breed has a comb known as the ‘walnut’ comb. Each of these breeds true.
Crosses between rose-combed and single-combed types show that rose is dominant to
single comb and that there is a segregation of 3 rose : 1 single comb in the F . In matings
2

between pea-combed and single-combed birds, pea comb is found to be dominant over single comb
and a 3 : 1 ratio appears in the F .
2

When, however, a rose-combed fowl is crossed with a pea combed one, all the F birds
1

show the walnut comb. When the F walnut combed birds are bred together, there appears in the
1

F 9 walnut : 3 rose : 3 pea : 1 single comb.

2

These results can be interpreted as follows: The rose comb is due to a gene R and the pea
is due to another gene P. The walnut comb is due to the presence of both the dominant genes, R
and P and the single comb is due to their recessive alleles, r and p.

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The above example depicts a case of non-epistatic intergenic interaction in which two genes that
determine the same character produce a new phenotype by mutual non-epistatic interaction.

Epistasis: (Non-allelic interaction) Interaction between two genes.

Expression of one gene masks the expression of the other is called epistasis

A gene which suppressing the other character is known as epistatic gene

Gene expression which was suppressed by a epistatic gene is known as hypostatic gene
The term epistasis is used for any kind of gene interaction.

Types of Epistasis

The various types of epistatic gene interaction include

1. Dominant epistasis or Simple epistasis (12:3:1)
In Dominant Epistasis, the dominant allele at one locus mask the expression of both
dominant and recessive alleles at another locus resulting in 12 : 3 :1 ratio
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2. Recessive epistasis or Supplementary gene action (9:3:4)
In Recessive Epistasis, the recessive alleles at one locus mask the expression of both
dominant and recessive alleles at another locus resulting in 9 :3 : 4 ratio
3. Dominant and recessive epistasis or Inhibitory gene action (13:3)
In this type of Epistasis, the dominant and recessive alleles at one locus mask the
expression of both dominant and recessive alleles at another locus resulting in 13 : 3 ratio
4. Duplicate recessive epistasis or Complementary gene action (9:7)
In this epistasis, the recessive alleles at either of the two loci mask the expression of
dominant alleles at the two loci, resulting in 9 : 7 ratio
5. Duplicate dominant epistasis or Duplicate gene action (15:1)
In this epistasis, the dominant alleles at either of the two loci mask the expression of
recessive alleles at the two loci, resulting in 15 : 1 ratio
6. Duplicate genes with cumulative effect or polymeric gene action (9: 6 : 1)
In this type of epistasis, two dominant alleles have similar effect when they are separate
but produced enhanced effect when they are together, resulting in 9 : 6 : 1

Difference between Dominance and Epistasis

Sl.No Dominance Epistasis

1. Interaction of two alleles of the same Interaction of two or more genes, thus
gene, thus involving single locus involving two or more loci
2. Always refers to Heterozygotes Refers to homozygotes and heterozygotes
therefore, it is not fixable therefore, it is fixable in homozygotes
3 Dominance is of three types viz., Epistasis is of several types viz., dominance,
complete, incomplete and duplicate and recessive
overdominance
4. Partial dominance alters the normal It modifies the normal dihybrid ratio in F2
segregation ratio of 3:1 into 1:2:1
5. It is known as intragenic or intralocus It is known as intergenic or interallelic or
gene interaction interlocus gene interaction
6. Recessive genes can express only in Recessive genes can also exhibit masking

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 homozygous condition effect

Dominant epistasis: (12:3:1)

Two genes affecting the same character produce distinct phenotypes when they are alone.
But when both the genes are present together, the expression of one gene masks the expression of
the other.

In Barley seed coat color is governed by two dominant B and Y

B - Black b- white

Y - Yellow Y - white

When both B_ Y_ present both the gene produce their effect but the B- Black colour is so intense
and it masks the expression of Y gene.

9 - B–Y- Black

3 - B-yy Black

3 - bb Y- Yellow

1 - bb yy White

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 When two non allelic genes affect the same trait of an organism, expression of one
dominant gene mask the expression of another gene. A gene thus mask the expression of another
gene is called epistatic gene and the gene hidden is called hypostatic gene.

Lecture 14. ii.) Recessive epistasis(9:3:4) iii.) Duplicate and additive epistasis((9:6:1). iv.)
Duplicate dominant epistasis(15:1)

Recessive epistasis: (9:3:4) (supplementary factor)

In this gene interaction, the dominant allele of one of the two genes governing a character
produces a phenotypic effect. However, the dominant allele of the other gene does not produce a
phenotypic effect of its own, but when it is present with the dominant allele of the first gene it
modifies the phenotypic effect produced by that gene.

Eg:- Maize grain colour / Aleurone colour.

RRPP x rrpp RR – Red

(purple) (white)

RrPp purple (RR – red PP – no.

colour when it (pp)
3P – 9 R-P - purple
combine with
3R- 1pp – 3R –pp – Red
another Dominant
3 P – 3 rr P_ gene (RR) it modify

1 rr White its phenotype)

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 1 p - 1 rr pp

Phenotypic ratio 9:3:4

Duplicate and additive epistasis: (9:6:1)

In this gene action, the two genes controlling a character produce identical phenotypes
when they are alone. But when the genes are present together, their phenotypic effect is enhanced
as if the effect of the two genes were cumulative (or) additive.

Eg:- In barley, two completely dominant genes A and B affect the length of awns. Gene
A or B alone gives rise to awns of medium length. But when both the genes A and B are present
together, they produce long awns indicating that the effects of A and B are added together.

Parents AA BB x aa bb

Long awn awnless

AaBb long awn.

F2

3B_ 9A- B - Long awn

3A- 1 bb 3A - bb

3B- 3 aa B - Medium

1 aa

1 bb aa bb awnless

Duplicate dominant interactions : (15: 1)

Characters showing duplicate gene action are determined by two completely dominant
gene these dominant genes produce the same phenotype, whether they are alone (or) together.

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The contrasting phenotype is produced only both the genes are in the homozygous recessive
state.

Eg: Non floating habit in rice is controlled by two dominant genes DW1, and Dw2. genes

DW1 and DW 2 alone, as well as together produce the same phenotype viz., non floating. The

floating habit is obtained only when both these genes are in recessive state.

Parents

DW1 DW 2 DW2 x dw1 dw1 dw2 dw2

(Non floating) floating

3Dw2 - 9 DW 1 _ Dw2_

N. floating

3 DW1-- 1dw2 - 3 DW 1 _ dw2 dw

3Dw2 - 3 dw1dw1 Dw2-

1 dw1dw1

1dw2dw2- 1 dw1 dw 1 dw2 dw2 - floating

Phenotypic ratio 15:1

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Lecture 15. v) Duplicate recessive epistasis (9:7) vi.) Dominant and recessive epistasis(13:3);
Summary of epistatic ratios (i)to (vi).

Duplicate recessive epistasis: ((9:7) complementary factor.

In this type of gene interaction, the production of one of the two phenotypes of a trait
requires the presence of dominant alleles of both the genes controlling the concerned traits when
any are of the two or both the genes are present in the homozygous recessive state, the contrasting
phenotype is produced.

Eg: In sweet pea, the development of purple flowers requires the presence of two dominant
genes C and R when either C or R or both the genes are present in recessive condition. Purple
colour flower cannot be produced as a result of which white flowers are obtained.

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 Parents CC RR x cc rr
(Purple) (white)
Cc Rr (purple)

F2 3R _ 9 C _ R _ Purple

3C_

rr 3C _ rr

1 cc 3 R _ 3 cc R _ White

1 rr 1 cc rr

Phenotypic ratio (9:7)

Dominant and recessive epistasis (13:3)- Inhibitory epistasis

In this gene action, one of the two completely dominant genes produces concerned phenotype
or the character while its recessive allele in homozygous state produces contrasting phenotypes.
The second dominant gene has no effect of its own. However it has the ability to stop the
expression of the first gene. As a result when the two dominant genes are present together, they
produce same phenotype as that produced by the recessive homozygous of first gene.

Eg: Development of aleurone colour in maize. A dominant gene ‘R’ produces red
colour, while its recessive allele ‘r’ reduces no colour. Another dominant gene ‘I’ does not
produce any colour by itself; it only prevents the color production by ‘R’ when both ‘I’ and
‘R’ are present together.

(i) Dominant Epitasis 12:3:1 Simple epistasis

(ii) Recessive Epitasis 9:3:4 (Supplementary factor)

(iii) Dominant & Recessive Epitasis 13:3 Inhibitory

(iv) Duplicate Recessive interaction 9:7 (Complementary)

v) Duplicate Dominant interaction 15:1 (Duplicate factor)

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 vi) Duplicate gene with cumulative effect 9:6:1 (Additive factor)

Lecture 16. Lethal genes, Pleiotrophy, penetrance and expressivity, phenocopy: Multiple
alleles, blood group in humans, coat colour in rabbits, self incompatibility in plants; pseudo
alleles, isoalleles.

Lethal Genes: Lethal genes are genes which in the homozygous state have such a marked
deleterious effect that such homozygous organisms are inviable. A lethal gene causes the death of
all the individuals carrying this gene in the appropriate genotype before these individuals reach
adulthood.
When seeds from self-pollinated maize plants are sown, sometimes green seedlings and
white seedlings (i.e., albinos) emerge in a ratio of approximately 3 green : 1 white. The albinos die
within a few says after germination and only green plants remain. The 3:1 ratio will be modified
into 2:1 .

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 Maize being a highly cross-pollinated crop is likely to be heterozygous. If the gene for
chlorophyll is represented as W and its recessive allele for albinism as w, the plant is likely to have
the genotype Ww. When this is selfed, homozygous recessives appear and these die.

Green plants Ww
Selfed
WW Ww Ww Ww
Green Green Green White (die)
Selfed Selfed Selfed
Green only Green and white Green and white

Seedlings of genotype ww always die after a short time in the field because they have no
chlorophyll which is absolutely essential for plants.
Lethal genes may be grouped into the following five categories.
1. Recessive Lethals
Most of the lethal genes are recessive lethal since their lethal effect is expressed only when
they are in the homozygous state and the survival of the heterozygote is unaffected.
Eg. In mouse hydrocephaly is due to a recessive lethal gene in homozygotes (recessive)
the gene causes abnormal growth of cartilage during embryonic development. This leads to
irregularly formed skull and brain and accumulation of cerebrospinal fluid. Such homozygotes do
not survive while their heterozygotes are apparently normal.

2. Dominant lethal:
Dominant lethal genes are lethal in homozygous conditions and produce some defective or
abnormal phenotype in heterozygous condition. Their most serious effect in heterozygotes may
also cause death.
Eg: Yellow lethal in mice
When ever the yellow males crossed with yellow females always yellow and brown were
obtained in the ratio of 2:1
Yy x Yy
(Yellow) (Yellow)
1 YY : 2Yy: 1yy

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 Die as : 2 yellow: 1 brown
Embryo
3. Balanced lethal:
Lethal genes can be used in establishing balance lethal system, where both homozygous
dominant and homozygous recessive die leaving only heterozygotes. Such balanced lethal systems
are known in Oenothera , Drosophila etc.,
4. Conditional lethal:
The genes which may be normal to the individual in a particular environment may prove
to be lethal when environment is changed.

Eg: - In Drosophila pseudoobscura. Flies live normally at a temperature at 16.5 0 C when

it raised to 250C, the flies begin to die.

5. Gametic lethal:
Some genes leads to the in-viability of a class of gametes or make them incapable of
fertilization such genes are termed as gametic lethal.
Eg: A dominant Segregation Distarter( SD) gene is known in the chromosome II of
Drosophila melanogaster
Complete lethal genes cause death of the zygote, developing embryo, fully developed
embryo or the fully developed organism before reproductive stage as in the case of coat colour in
mice, seedling colour (albino) in maize, snapdragon and many other plants.

Pleiotrophy:
A single gene controlling more than one character of the organism ie. To say that a gene
produces a major phenotypic trait. But in addition to that influences some other phenotypic traits.
The phenomenon of multiple or manifold phenotypic expression of a single gene is called
Pleiotropism and such genes are known as pleiotropic genes.
Eg: (i) In cotton, the Punjab hairy lintless gene “lic produces seeds which are without lint.
This gene also causes incomplete lancination of leaf, reduction in number and length of internodes,
reduction in boll size and fertility.

84
(ii) In Drosophila, the recessive gene for vestigial wings also affects structure of reproductive
organs, reduction in egg production, reduction in longevity and the bristles on the wings.

(iii) In man, gene producing disease called Phenyl ketoneuria also produces number of abnormal
traits which are collectively known as syndrome other effects are short stature, mental
retardation, widely spaced incisors, pigmented patches on the skin and excessive sweating.
Penetrance
The ability of a gene or gene combination to be expressed phenotypically to any degree is
called penetrance.
The proportion of individuals of a specified genotype that show the expected phenotype
under a defined set of environmental conditions called penetrance. It is the ability of the gene to
express its character.
Complete penetrance
Most dominant and recessive genes in homozygous condition and many completely
dominant gene even in heterozygous condition give their phenotypic expression. Heterozygous
condition also produces the normal phenotype.
Incomplete penetrance:
Heterozygous condition unable to express fully the normal phenotype.
Eg: In the case of polydactyly in man where one extra finger is present in the palm or foot,
the heterozygous condition (Pp) brings forth polydactyly in some and normal condition in others.
Some heterozygotes individuals were not polydactyles some had an incomplete penetrance.

Expressivity:
The degree of phenotypic expression of a gene in the different individuals is known as
expressivity, ie; the degree of effect produced by the Penetrance genotype:
Change of temperature, nutrition, hormone deficiency etc., influence the expressivity of the
curly wing in Drosophila.
Eg:- In man polydactyl condition may be penetrance in left hand (6fingers) not in right
hand (5 fingers) or may be penetrance with the feet but not in the hand.

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Phenocopy:
The phenotype becomes altered by the environment in such a way that the new phenotype
resembles another phenotype produced by known genes. The induced phenotype is not inherited
and is called as phenocopy. Eg:- Generally the body colour of the fruit fly Drosophila
melanogaster is light brown. When the larvae of the normal brown bodied fruit flies were reared
on food with silver salts, the emerging adults had yellow body

Multiple alleles

More than two alleles at the same locus give rise to a multiple allelic series. Therefore, the
existence of more than two alleles at the same locus of a homologous chromosome is referred to
as multiple alleles
Presence of multiple alleles adds variability for a character
The number of possible genotypes in a series of multiple alleles can be calculated by using the
formula ½ {n x 1 (n + 1)} where, n is the number of identified alleles in that group. For example,
if there is 4 alleles in the multiple allelic series, then, ½ (4 x 5)=10 genotypes are possible

Main features of multiple alleles

1. Multiple alleles always belong to the same locus and one allele is present at a locus at a
time in a chromosome
2. Multiple alleles always control the same character of an individual. However, the
expression of the character will differ depending on the allele present.
3. There is no crossing over in a Multiple allelic series. When two alleles are involved in a
cross, the same two alleles are recovered in the F2 or test cross progeny. This is based on
the classical concept of the gene, according to which crossing over takes place between
gene and not within a gene.
4. In a series of Multiple alleles, wild type is always dominant. Rest of the alleles in the series
may exhibit dominance or intermediate phenotypic expression when two alleles are
involved in a cross.
5. The cross between two mutant alleles will always produce mutant phenotype
(intermediate). Such cross will never produce wild phenotype. In other words, Multiple
alleles do not show complementation (Complementation refers to appearance of wild
phenotype when two mutants are crossed)

Examples of Multiple Alleles

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 Several cases of multiple alleles are known to occur in both plants and animals. Some
well known examples for expression of multiple alleles include,
1. Fur color in Rabbits
2. Wing type in Drosophila
3. Eye colour in Drosophila
4. Self incompatability alleles in Plants
5. ABO Blood group in man

1. Fur colour in Rabbits

It is a well known example for multiple alleles
In rabbits, the fur colour is of four types, agouti, chinchilla, Himalayan and albino
1. Agouti:
 This has full colour, known as wild type
 This colour is dominant over all the remaining colours
 Produces agouti colour in F1 and 3:1 in F2 when crossed with any of the other
three colours in rabbits.
 This colour is represented by C
2. Chinchilla:
 This is lighter than Agouti.
 This colour is dominant over Himalayan and albino
 Produces chinchilla in F1 and 3:1 ratio in F2 when crossed with either Himalayan
or albino.
 This colour is represented by cch
3. Himalayan:
 The main body is white while the tips of ear, feet and tail and snout are coloured
 This colour is dominant over albino
 Produces 3:1 ratio in f2 when crossed with albino.
 This is represented by ch
4. Albino:
 This has pure white fur colour and is recessive to all other types.
 This is represented by c
Thus the fur colour in rabbits can be represented in the order of dominance as,
Agouti Chinchilla Himalayan Albino

(C) ( cch) (ch) (c)

Cross between Expression in F1 Segregation in F2

Agouti x chinchilla Agouti 3 Agouti : 1chinchilla

Agouti x Himalayan Agouti 3 Agouti : 1himalayan
Agouti x albino Agouti 3 Agouti : 1albino

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 Chinchilla x Himalayan Chinchilla 3 Chinchilla : 1Himalayan
Chinchilla x albino Chinchilla 3 Chinchilla : 1 albino
Himalayan x albino Himalayan 3 Himalayan : 1albino

Blood Groups in Human

The A-B-O blood group in human being controlled by three alleles IA, IB and IO to form
a multiple allelic services.

Blood Group Possible genotype

O IO IO
A IA IA, IA IO
B IB IB, IB IO
AB IA IB

IA and IB are codominant of each other and both are completely dominant over IO.

Pseudoalleles
Pseudoalleles are non alleles so closely linked as often inherited as one gene, but
shown to be separate by cross over studies.
One of the first demonstrations of pseudoallelic condition was that of star-asteroid analysed
by Lewis in 1951. He found a recessive mutation in Drosophila producing a small rough eye when
homozygous. It was at locus 1.3 in the second chromosomes. This was also the identical location
of gene star, a dominant mutation also affecting the morphology of the eye. The eye was rough and
had a slight gleam, hence the name star, with gene symbol S. In crosses among these flies,
recombination between star and asteriod occurred at a low frequency of one in five thousand.
Pseudoallelic effects are found in Drosophila, corn, cotton, Aspergillus, Neurospora,
bacteria and in viruses.

Isoalleles

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 Usually wild type allele (represented as +) is dominant over its recessive allele. In some
natural populations, different wild type alleles affecting the same character were found and these
wild type alleles had similar allelic dominance or they may differ in their degree of expression that
could be detected in special combinations. Such alleles are called isoalleles.
Timofeev - Ressovsky and Muller found that the wild type Drosophila from different
natural populations had different dominant (red eye) alleles as judged by their stability or by their
different effects in combinations. Stern found three different wild type alleles of another
Drosophila mutant, cubitus interruptus, which showed different degrees of dominance over the
same mutant allele. He called such alleles as isoalleles because they were alike in their
homozygous effect and their differences appeared only in special combinations.

Modifying genes
A modifying gene is one that alters the expression of a major gene but has no effect on the
allele of the major gene. The modifiers have very similar but individually small effects and
are usually present in such large numbers that they cannot be individually identified.

In the Guernsey breed of dairy cattle, the `solid' colour (fawn i.e., light yellowish brown) of the
coat is due to dominant gene S and the `spotted' coat (white spotting) is due to its recessive allele
s. A number of these modifying genes influence the intensity of spotting. If a large number of these
modifying genes is present in animals with ss, the animals are highly `spotted'. If only a small
number of these modifying genes is present in animals with ss, they are medium `spotted'. If the
modifying genes are absent, animals with ss have only few `spots'. These modifying genes have
no effect in the presence of the gene for `solid' colour and animals with SS or Ss have solid-
coloured coats irrespective of the number of modifyig genes present.
In Gossypium barbadense the presence of petal spot is due to a gene S and the absence of
petal spot is due to its recessive allele s. A number of modifying genes increases the intensity of
colour in the presence of the gene S.

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Lecture 18. Quantitative inheritance – Multiple factor hypothesis – Nilsson Ehle, his e
x p e r i m e n t
It is quite natural that small differences exist among individuals of similar genotype due to the
effect of environment on genotype.
• On the other hand, there are some heritable differences also exist with continuous variation.
• Most of the economical traits show continuous variation and they are measurable or
quantifiable.

Quantitative characters
• Quantitative characters are traits which show
• continuous variation and
• governed by a large number of genes called multiple genes or
• multiple factors or polymeric genes or polygenes.
• Their inheritance follows same mendelian principles.
Qualitative characters
Qualitative characters show
• discontinuous variation and
• are governed by one or two major genes or oligognes.

Multiple factor Hypothesis (Nilson – Ehle -1908) experiment on wheat kernel color

Multiple factor hypothesis (Nilson - Ehle)

i) for a given quantitative trait there could be several genes, which were independent in their
segregation, but had cumulative effect on phenotype
ii) Dominance is usually incomplete
iii) Each gene contributes something to the strength of expression of character whereas its
recessive allele does not of genes present dominance gene.

• Nilson-Ehle studied Kernel colour in wheat

• concluded that is a quantitative character

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 • He crossed true breeding red kernel whet (RR) with true breeding white (rr) and the F1 was
red (Rr) and the F2 segregated for red and white in 3:1 ratio indicating the dominance of
red over white.
• However, careful examination indicated the variation in red color among the red color
progenies
• F1 red was not as intense as one of the parents
• In F2 he could observe two grades of red ie., one was red as that of one of its parent, two
were higher red as that of F1 individuals.
• In some crosses, a ratio of 15 red : 1 white was found in F2 indicating that there are two
pairs of genes for red colour that either or both of these can produce red kernels.
 Finally he observed different shades of red in F2 for red kernel types. The F2
showed red shades and white as follows;
Dark red : 1
• Medium dark red : 4
• Medium red : 6 15
• Light red : 4
• White : 1
Total : 16
• It was concluded two duplicate dominant alleles R1 and R2 cumulatively decide the
intensity of red colour
• and both R1 and R2 are in completely dominant over white.
• The high intensity of red colour depends on the number.
The F2 ratio in wheat
Genotype Genotypic ratio Phenotype
R1R1 R2R2 1 Dark red
R1R1 R2r2 2 Medium dark red
R1r1 R2R2 2 Medium dark red
R1r1 R2r2 4 Medium red
R1R1 r2r2 1 Medium red
r1r1 R2R2 1 Medium red
R1r1 r2r2 2 light red
r1r1 R2r2 2 light red

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 r1r2 r2r2 1 white

• Hence, if two parents differ for the two genes the segregation was 1:4:6:4:1 provided both
R1 and R2 contribute equally to the colour.
 If three genes are involved in F2 segregation showed 1:6:15:20:15:6:1 for red
shades and 1 for white.

Multiple factor –Quantitative inheritance

The alleles that contribute for the trait are called "contributing alleles",

If four genes are determining the kernel color then,

R1R1 R2 R2 x r1r1 r2r2
dark red white

F1 R1r1 R2r2
Medium red

♂ R1R2 R1r2 r1R2 r1r2

♀
R1R2 R1R2 R2R2 R1R1 R2r2 R1r1 R2R2 R1r1 R2r2
(dark red) (Medium dark red) medium dark red light red

R1r2 R1R1 R2r2 R1R1 r2r2 R1r1 R2r2 R1r1 r2r2

medium dark red light red light red very light red
r1R2 R1r1 R2R2 R1r1 R2r2 r1r1 R2R2 r1r1 R2r2
medium dark red light red light red very light red
r1r2 R1r1 R2r2 R1r1 r2r2 r1r1 R2r2 r1r1 r2r2
light red very light red very light red white

Dark red is due to the presence of four contributing genes medium dark is due to three
genes and medium red is due to two contributing genes and light red is due to one contributing
gene

Quantitative inheritance in ear length of Maize (Emerson & East)

• In maize, the ear (cob) length is governed by multiple factors.
• Two varieties of maize viz., long eared sweet corn and short eared popcorn.
• The earlength of sweet corn ranges from 13-21 cm with an average of 16.8 cm.
• While that of pop corn was ranging from 5 to 8 cm with an average of 6.6 cm when these
varieties were crossed,
• the F1 progeny was found to have intermediate size ranging from 9-15 cm with an
average of 12.1cms.
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 • In F2 there were relatively few of these extreme and relatively large number of these
extreme and
• relatively large number of these with intermediate ear size which implies that ear size
differences between two parents may be due to one, two or several gene differences.

Similarly in tobacco (Nicotiana longiflora) the corolla length shows polygenic inheritance.

Lecture 19. Polygenes – transgressive segregation, comparison of quantitatively and

qualitatively inherited characters; modifiers; Types of gene action controlling quantitative
traits

Polygenes: Any of a group of nonallelic genes, each having a small quantitative effect, that
together produce a wide range of phenotypic variation. Also called multiple factor, quantitative
gene.
Transgressive segregation
When the range in the F2 progeny goes beyond the original parents, it is known as
transgressive segregation ". Transgressive segregation implies that the parents donate contributing
alleles from different genes to the hybrid .
The appearance of individuals in F2 with very higher or lower intensity of expression than their
both parents is known as transgressive segregation and such individuals are called transgressive
segregants.
Transgressive segregants are produced when

i) the two parents involved in a cross which have positive alleles of different
genes affecting a quantitative characters.
Segregation for these genes produces the two extreme homozygotes in F2 which transgress the
parental limits for the character
.Quantitative inheritance for skin color in human beings

93
 Genotypic variance

Heritability =
Total or phenotypic variation

The fraction of the total variance that is due to genotype is called in the broad sense.

The differences between qualitative and quantitative traits

Qualitative traits Quantitative traits
1. Segregation of alleles cause differences Segregation of contributing genes causes
in phenotypic character differences in phenotyeic characters
2. Qualitative traits are governed by Oligo Quantitative traits are governed by polygenes or
genes or mendelian genes multiple factors
3. Effect of each allele is easily effect of individual contributing genes is not
distinguishable distinguishable
4. discrete variation continuous variation
5. Traits are classified by more counting quantative traits are measured on a continuous
scale (mean, variance) with statistical method)
6. Influence of environment is very less More influence of environment
7. Dominance is complete in mendelian each dominant gene has a quantitative effect
ratio
8. Lesser role of modifying genes more role of modifying genes

Features of quantitative inheritance or polygenic inheritance

1. The trait is governed by polygenes
2. segregation occurs at an indefinitely large number of loci

94
3. Polygenes are always under the influence of environmental factors and hence the
quantitative trait is altered.
4. Multiple genes are usually incompletely dominant duplicate genes with cumulative
effect
5. The phenotypic differences (variation) within a parental line or F1 population are
purely due to environment
6. The F1 individuals have the same genotype and they are intermediate between parents
7. The F2 exhibits considerable variability. The variation within individuals is mainly
due to genotype and the variation is continuous.
8. The F2 mean is equal to the F1 mean and the parental mean
9. The individual effect of individual contributing genes can not be distinguished early
when compared to the environmental influence
10. The estimates of quantitative inheritance are calculated with mean, variance,
heritability and genetic advance
11. They are measurable characters height of 100 cm. The recessive alleles t1t1t2t2 do
not influence the height of plant. T1 and T2 due incompletely dominant over t1 and
t2 respectively.

95
 Lecture 20. Linkage - coupling and repulsion; Experiment on Bateson and Punnet –
Chromosomal theory of linkage of Morgan – Complete and incomplete linkage, Linkage
group

LINKAGE AND CROSSING OVER

Bateson and Punnett discovered in 1906 that the principle of independent assortment of
members of different pairs of alleles at the timeof formation of gametes is not universsal but has
same exception. Thomas Hunt Mongan (1910) found similar situations in Droshphila to give a
satisfactory explanation for such deviation.

Linkage
• The tendency of two or more genes to stay together during inheritance is known as
linkage
• Linked genes do not show independent segregation
• Group of genes situated on the same chromosome is known as linkage group
• The number of linkage group = the number of haploid chromosome number
• Any two genes on a linkage group is known syntenic

96
 According to 'Chromosome Theory of Inheritance,, the genes are carried in the chromosomes.
But the number of genes per individual far exceeds the number of chromosome pairs. It means
each chromosome bears many genes. The genes located on the same chromosome cannot assort
independently, rather these tend to be inherited together. This phenomenon of inheritance of genes
together and to retain their parental combination even in the offsprings is known as linkage. The
genes located in the same chromosome and being inherited together are known as linked genes,
and the characters controlled by these are linked characters.

The difference between linkage and independent assortment can be illustrated

by the following figure
i

97
 All those genes which are located in the single chromosome! constitute a linkage
group. The total number of linkage groups in mu organism is equal to the number of chromosome
pairs. For example, there are 4 linkage groups in Drosophila melanogaster, 23 in man and 7 in
sweet pea.

HISTORY OF LINKAGE
The theory of linkage was propounded by T. H. MORGAN in 1911. But
its existence was predicted even before that and was described under!- different
names.
1. Sutton's Hypothesis

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 Sutton (1903) suggested that each chromosome bears more than one genes and all the
genes, situated in one chromosome are inherited together in the offsprings, but he was unable to
support his hypothesis experimentally.

2. Coupling and Repulsion Hypothesis

The event of linkage was first observed by BATESON and PUNNET in 1906 in pea plant,
but it was described as coupling. They found that the results of dihybrid cross in sweet pea,
Lathynis, odoratus, involving colour and shape of pollen grains, do not agree with the law of
independent! assortment. The results obtained are shown in the table below

P1 Purple flower; long pollen x Red flower, round pollen

F1 Purple flower; long pollen

Generation Phenotype Number Ratio

F2 Purple long 296 11

Purple round 29 1
Red long 27 1
Red round 85 3

When these F1 purple, long (heterozygous) hybrids were crossed with the double recessive

red and round )homozygous) individuals (test cross) failed to produce expected 1: 1: 1: 1 ratio in
F2 generation. These actually produced following four combinations in the ratio of 7:1:1:7.

P1 Purple long x Red round

RRLL rrll

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 F1 Purple long x Red round (double recessive)

RrLl rrll

Test cross
Progeny
Purple long Purple round Red long Red round
7 1 1 7
7 purple long : 1 purple round:1 red long : 7 red round

The above results of the test cross indicate that the parental combinations are seven times more
numerous than the non-parental combinations. Bateson and Punnet suggested that the alleles
coming from the same parent tend to enter the same gamete and to be inherited together (genetic
coupling). Similarly, the same genes coming from two different parents tend to enter different
gametes and to be inherited separately and dependently (repulsion).

Bateson and Punnet could not explain the exact reasons for coupling and repulsion. Although,
the theory is obsolete now, the terms ‘coupling phase’ and 'repulsion phase' have been retained on
account of heir descriptive significance.

3. Morgan's Concept of Linkage

Morgan (1910) while working on Drosophila stated that coupling and repulsion are two
aspects of the same phenomenon, which he described as 'linkage'. He defined linkage 'the tendency
of the genes, as present in the same chromosome, to remain in their original combination and to
enter together in the same gamete.

4. Chromosome Theory of Linkage

Morgan and Castle formulated The chromosome Theory of linkage. It has following
characteristics: —
1. Genes that show linkage are situated in the m chromosome.
2. Genes are arranged in a linear fashion in the chromosome. Linkage of genes is linear.

101
 3. The distance between the linked genes is inversely proportional to the strength of
linkage. The genes which are closely located show strong linkage, whereas those,
which are with separated, have more chances to get separated by cross over.
4. Linked genes remain in their original combination during the course of inheritance.

The chromosome theory of linkage is widely supported from the cytological studies. It has
helped in the construction of linkage maps of chromosomes (The distance between the genes is
determined by the percentage of cross overs.

ARRANGEMENT OF LINKED GENES

In an individual, which is heterozygous for two pairs of linked genes, the linkage can be
either of the two types:-
(i) The dominant genes of both the pairs are located in one member of the chromosome
pair and their recessive alleles are located in the other chromosome of the pair.
This arrangement is known as cis-arrangement. And the heterozygous with such
arrangement (AB/ab) are known as cis - heterozygotes –COUPLING PHASE
(ii) The dominant gene of one pair and the recessive gene of other pair are located on one
chromosome of the pair and the recessive gene of the first pair and dominant gene of the
second pair are located in the second chromosome pair (Ab/aB). This arrangement of a
dominant and recessive gene in the same chromosome of the chromosome pair is known as
rans-arrangement and heterozygotes with such arrangement are called trans-heterozygotes.
REPULSIVE PHASE

102
 A
Diagram exhibiting the trans and cia
a
arrangement of genes
b

Types of linkage
Completely linked genes: any two
a
genes on a A p a r t i c u l ar
chromosome is very close to each other
B
and move together b to the gametes and
n o recomination/cross
overs/chiasma between these two loci. Eg. Male drosophila
Incompletely linked – any two genes on the same chromosome but show moderate level of cross
overs
Tightly linked genes – show very little frequency of recombination

Advantages of linkage
• Linkage provides a way to map genes on chromosomes. (Until modern gene mapping
techniques linkage was the only way to do this)
• .Even today linkage is important when used with other techniques to determine the
location of genes and also for diagnosis of disease
• the presence of linkage complicate the evolutionary behavior of genes in populations

Lecture 21. Crossing over – significance of crossing over; cytological proof for crossing
over - Stern’s experiment; Factors controlling crossing over.

Crossing over
Crossing over is the exchange of strictly homologous segments between non sister chromatids of

103
homologous chromosomes during pachytene stage of prophase I of meiosis I.

104
Each event of crossing over produces
− two recombinant chromatids (involved in the crossing over) – crossover
chromatid
− Two original chromatid (non cross over chromatid)

The term crossing over was first used by Morgan and Cattell in 1912.
The main features of crossing over are given below :
1. Crossing over takes place during meiotic prophase, i.e., during pachytene. Each pair of
chromosome has four chromatids at that time.
2. Crossing over occurs between non-sister chromatids. Thus one chromatid from each of
the two homologus chromosomes is involved in crossing over.
3. It is universally accepted that crossing over takes place at four strand stage.
4. Each crossing over involves only two of the four chromatids of two homologus
chromosomes. However, double or multiple crossing over may involve all four, three or
two of the four chromatids, which is very rare.
5. Crossing over leads to recombinations or new combinations between linked genes.
Crossing over generally yields two recombinant types or crossover types and two parental
types or non-crossover types.
6. Crossing over generally leads to exchange of equal segments or genes and recombination
is always reciprocal. However, unequal crossing over has also been reported.
7. The value of crossover or recombinants may vary from 0-50%.
8. The frequency of recombinants can be worked out from the test cross progeny. It is
expressed as the percentage ratio of recombinants to the total population (recombinants +
parental types) Thus,
No. of recombinants
Crossing over frequency (%) = -------------- x 100
Total progeny

Differences between crossing over and linkage

Crossing over Linkage

1. It leads to separation of linked genes. It keeps the genes together.

105
 2. It involves non-sister chromatids of It involves individual chromosome.
homologous chromosomes.
3. Frequency of crossing over can never Linkage groups can never be more than
exceed 50%. haploid chromosome number.
4. It increases variability by forming new gene It reduces variability.
combinations.
5. It provides equal frequency of parental and Provides higher frequency of parental
recombinant types in test cross progeny. types than recombinant types in test
cross progeny.

Chiasma and Crossing over

The point of exchange of segments between non-sister chromatids of homologous
chromosomes during meiotic prophase is called chiasma (pleural chiasmata). It is thought to be
the place where crossing over takes place. Chiasma was first discovered by Janssens in 1909.
Depending on the position, chiasma is of two types, viz., terminal and interstitial. When the
chiasma is located at the end of the pairing chromatids, it is known as terminal chiasma and when
it is located in the middle part of non-sister chromatids, it is referred to as interstitial chiasma.
Later on interstitial chiasma is changed to terminal position by the process of chiasma
terminalization. The number of chiasma per bivalent may vary from one to more than one
depending upon the length of chromatids. When two chiasmata are formed, they may involve two,
three or all the four chromatids.

Chiasma Terminalization
The movement of chiasma away from the centromere and towards the end of tetrads is called
terminalization. The total number of chiasmata terminalized at any given stage or time is known
as coefficient of terminalization. Generally, chiasma terminalization occurs between diplotene and
metaphase I.
TYPES OF CROSSING OVER
Depending upon the number of chiasmata involved, crossing over may be of three types,
viz., single, double and multiple as described below :

Single Crossing Over

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 It refers to formation of a single chiasma between non-sister chromatids of homologous
chromosomes. Such cross over involves only two chromatids out of four.
Double Crossing Over
It refers to formation of two chiasmata between non-sister chromatids of homologous
chromosomes. Double crossovers may involve either two strands or three or all the four strands.
Multiple Crossing Over
Presence of more than two crossovers between non-sister chromatids of homologous
chromosomes is referred to as multiple crossing over. Frequency of such type of crossing over is
extremely low.

CYTOLOGICAL PROOF OF CROSSING OVER – STERNS EXPERIMENT ON

DROSOPHILA –CYTOLOGICAL PROOF FOR CROSSING OVER

The first cytological evidence in support of genetic crossing over was provided by Curt Stern
in 1931 on the basis of his experiments conducted with Drosophila. He used cytological markers
in his studies. He selected a female fly in which one X-chromosome was broken into two segments.
Out of these two segments, one behaved as X-chromosome. The other X-chromosome had small
portion of Y-chromosome attached to its one end. Thus, both the X-chromosomes in the female
had distinct morphology and could be easily identified under microscope. In female fly, the broken
X-chromosome had one mutant allele (carnation) for eye colour and another dominant allele (B)
for bar eye shape. The other X-chromosome with attached portion of Y chromosome had alleles
for normal eye colour (red eye) and normal eye shape (oval eye). Thus, phenotype of female was
barred.

A cross of such females was made with carnation male (car+). As a result of crossing over
female flies produce four types of gametes, viz., two parental types or non crossover types (car B
and ++) and two recombinant types or crossover types (car+ and B+). The male flies produce only
two types of gametes (car+ and Y), because crossing over does not take place in Drosophila male.
A random union of two types of male gametes with four types of female gametes will produce
males and females in equal number, means there will be four females and four males

107
108
 Stern examined the chromosomes of recombinant types, viz., red bar and carnation normal
under microscope. He observed that in carnation normal females both the X-chromosomes were
of equal length.
In red bar flies, one X-chromosome was normal and other was fragmented. The fragmented
X-chromosome also had attached part of Y-chromosome. Such chromosome combination in red
bar is possible only through exchange of segments between non-sister chromatids of homologous
chromosomes. This has proved that genetic crossing over is the result of cytological crossing over.
Similar proof of cytological crossing over was provided by Creighton and McClintock in maize.
Cytological evidence for crossing over in the autosomes (chromosomes other than sex
determining chromosomes) is available in maize (n = 10), in which Creighton and McClintock
(1931) identified a mutant. The mutant had a satellite (knob) at the terminal end of the ninth
chromosome, which also had a segment of the eighth chromosome translocated to it. The genes

for coloured or colourless aleurone and starchy or waxy endosperm were located on the 9 th
chromosome. Recombinations due to crossing over were recognized in the progeny of a cross

between the female parent having the aberrant 9 th chromosome with its normal homologue and a

male parent with normal 9th chromosome. The crossover could be cytologically detected, thus
establishing the cytological basis for genetic crossing over.
FACTORS AFFECTING CROSSING OVER
The frequency of crossing over is influenced by several factors which are briefly discussed
below :
1. Distance, The distance between genes affects the frequency of crossing over. Greater the
distance between genes higher is the chance of crossing over and vice versa.
2. Age. Generally crossing over decreases with advancement in the age in the female
Drosophila.
3. Temperature. The rate of crossing over in Drosophila increases above and below the
temperature of 22°C.
4. Sex. The rate of crossing over also differs according to sex. There is lack of crossing over in

109
 Drosophila male and female silk moth.
5. Nutrition. Presence of metallic ions like calcium and magnesium in the food caused
reduction in recombination in Drosophila. However, removal of such chemicals from the
diet increased the rate of crossing over.
6. Chemicals. Treatment with mutagenic chemicals like alkylating agents was found to increase
the frequency of crossing over in Drosophila female.
7. Irradiation. Irradiation with X-rays and gamma rays was found to enhance the frequency of
crossing over in Drosophila females.
8. Structural Changes. Structural chromosomal changes especially inversions and
translocations reduce the frequency of crossing over in the chromosomes where such changes
are involved.
9. Centromere Effect. Generally genes that are located adjacent to the centromere show
reduced frequency of crossing over.
9. Cytoplasmic Genes. In some species cytoplasmic genes also lead to reduction in crossing
over. For example, Tifton male sterile cytoplasm in pearl millet.

SIGNIFICANCE OF CROSSING OVER

Crossing over is useful in three principal ways, viz., (1) creation of variability, (2) locating
genes on the chromosomes, and (3) preparing linkage maps as described below:
1. Creation of Variability: Crossing over leads to recombination or new combination and thus
is a potential genetic mechanism for creating variability which is essential for improvement
of genotypes through selection.
2. Locating genes : Crossing over is a useful tool for locating genes in the chromosomes.
3. Linkage maps : Crossing over plays an important role in the preparation of chromosome
maps or linkage maps. It provides information about frequency of recombination’s and
sequence of genes which are required for preparation of linkage maps.

110
Lecture 22. Strength of linkage and recombination; Two point and three point test cross.

Linkage studies revealed the following:

1. Genes that assort at random are unlinked genes. Genes that do not segregate at random are
linked genes. They are situated in the same pair of chromosomes and are transmitted in
unitary groups.
2. The linked genes are arranged in a linear fashion on the chromosome. Each linked gene
has a definite and constant order in its arrangement.
3. The distance between the linked genes determines the degree of strength of linkage.
Closely located genes show stronger linkage than the widely located genes.

Two-point Test Cross -A cross of dihybrid with its homozygous recessive parent
Three point testcross: A three point test cross is a cross of a trihybrid (Ft differing in three genes)
with its homozygous recessive parent. The three point test cross provides useful information on
two important aspects, viz., (1) about the sequence of genes, and (2) about the recombination
frequencies between genes. This information is essential for mapping of chromosomes.

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Two-point Test Cross
Estimation of strength of linkage from test cross data
The value of linkage can be determined from the test cross data by dividing the total
number of individuals with the parental combinations divided by the total number of progeny and
multiplying it by 100.
Let us calculate the value of linkage from the results of the test cross:
P Coloured full X Colourless shrunken
CS/CS Cs/cs
F1 Coloured Full
CS/cs
Test cross Coloured full X Colourless shrunkle
CS/cs Cs/cs

Test cross progeny

Coloured full CS/cs 4032
Coloured shrunken Cs/cs 149
Colourless full CS/cs 152
Colourless shrunken Cs/cs 4035
Total number of progeny 8368
Parental combinations
Coloured full 4032
Colourless shrunken 4035
Total 8067
Percentage 8067/8368 * 100 = 96.4
Recombinations
Coloured shrunken 149
Colourless full 152
Total 301

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 Percentage 100 - 96.4 = 3.6
Linkage value 96.4%
Cross over value 3.6%

Estimation of linkage from F2 data

Linkage values can be calcualted from the F2 data by the additive method or the product

method or the maximum likelihood method.

We shall now illustrate the calculation of the linkage value from the F2 data.

Assume that a cross CS/CS * cs/cs gives in F2

Coloured full (a) 7300

Coloured shrunken (b) 200
Colourless full © 200
Colourless shrunken (d) 2300

i) Additive method:

The formula applied is p2 = E - M / N

Where p = linkage value; E = sum of end classes; M = sum of middle classes and N =
total number of progeny.

P2 = (7300 + 2300) - (200 + 200) / 10000

P2 = 0.92
P = 0.9592
Linkage value = 95.92%
Cross over value = 4.08%
ii) Product ratio method:
Product ratio (P) = ad / bc

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 P2 = (P + 1) - root (3P + 1) / P - 1
Where P = linkage value,
P= product ratio,
A and d=No. of F2 progeny in the parental combinations and b and c = No. of F2 progeny in the

recombinations.
Substituting the figures,
P = 7300 * 2300 / 200 * 200 = 419.75
P2= (419.75 + 1) - root (3 * 419.75) - 1 / 419.75 - 1

= 0.920072
p = 0.9592 1 - p = 0.0408
Linkage value = 95.92%
Cross over value = 4.08%
iii) Square root method:
Percentage of non-cross over gametes = Frequency of double recessives
= square root(2300 / 10000)
= 0.9592
Linkage value = 95.92%
Cross over value = 4.08%

Chiasmata
• The points at which the chromosomes actually cross over are called chiasmata (singular
chiasma),
• They involve large, multi-enzyme complexes that cut and join the DNA.
• There is always at least one chiasma in a bivalent, but there are usually many, and it is
the chiasmata that actually hold the bivalent together.
• The chiasmata can be seen under the microscope and they can give the bivalents some
strange shapes at prophase I.
• There are always equal amounts crossed over, so the chromosomes stay the same length.

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 • Ultimately, crossing over means that maternal and paternal alleles can be mixed, even
though they are on the same chromosome i.e. chiasmata result in different allele
combinations.

The probability of chiasma occurring between two linked genes

• Chiasma % = 2 (crossover %)
− or
• Crossover % = 1/2 (chiasma %)
• Chiasma frequency is twice the crossover frequency because when a chiasma forms
between two genes, only half of the meiotic products will be of the crossover type.

Lecture 23. Double cross over, interference and coincidence; genetic map, physical map.

Genetic map(cross over map or chromosome map or Linkage map)

CHROMOSOME MAPPING
Chromosome map refers to a line diagram which depicts various genes present on a
chromosome and recombination frequency between them. Such maps are also known as genetic
maps or linkage maps. The process of assigning genes on the chromosomes is known as

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chromosomal mapping. The mapping of chromosomes is done with the help of three point test
cross. A three point test cross is a cross of a trihybrid (Ft differing in three genes) with its

homozygous recessive parent. The three point test cross provides useful information on two
important aspects, viz., (1) about the sequence of genes, and (2) about the recombination
frequencies between genes. This information is essential for mapping of chromosomes.
Morgan postulated that genes are arranged in a linear order along the length of the
chromosome, each gene having a fixed place on the chromosome and its allele, a corresponding
position on the homologous chromosome.
In mapping genes, a unit of distance must be used and this unit is called a map unit which
is the space within which one per cent of crossing over takes place. If the percentage of cross over
between two linked genes is 1 per cent, it means that the map distance between these two linked
genes is one unit of map distance or one map unit, or one centimorgan.
The chromosome map may be defined as a line, on which the genes are represented by
points, separated by distances proportional to the amount of crossing over.
The chromosome maps are also referred to as cross over maps since they are sketched by
the amount of crossing over.
The percentage of crossing over is directly proportional to the distance of the alleles
showing crossing over in the chromosome.
The chromosomes maps are the graphic representation of the genes in a chromosome.
The percentage of crossing over is calculated by test crosses. In mapping the genes, a unit
of distance is used and it is called as map unit or Morgan unit.
The first chromosome map was made in 1911 by Sturtevant and soon after additional maps
were made by Bridges and others.
Drosophila is the earliest material used by the scientists, for constructing maps.
Procedure for the chromosome mapping
In fact genes are plotted on the chromosome on the basis of crossing over results between
different pairs of linked genes. The actual distance between two genes is said to be equivalent to
the percentage of crossing over between these genes.
When the % of crossing over between two genes is 5, then the distance is 5 units. For
example five genes A, B, C, D and E are to be plotted on a chromosome. If cross over results

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indicate that genes A and E have the highest percentage of crossing over, it means that these should
be placed at the maximum distance.
In this example, the gene A can be taken as a starting point in the chromosome and can be
represented by O.
Now if the gene A and B exhibit 7% crossing over, the gene B can be placed on the
chromosome at a distance of 7 units.
If the gene C shows 8% crossing over with gene B and about 15% crossing over with gene
A, it can be plotted on the chromosome at a distance of 15 units from gene A.
Similarly if gene A and E exhibit 20% and 30% crossing over with gene D and 5% and
10% with gene C these, are located on the chromosome 5 and 10 units away from the gene C
respectively.
Construction of Chromosome map in Drosophila
In Drosophila the chromosome map is constructed with the help of test cross. In Drosophila
grey colour is dominant over black colour; and the long wing is dominant over vestigial wing.
The F1 female hybrid is test crossed. Four types of individuals are formed. Out of four

types, two types are parental type (G:L & B:V) and other two are non parental type (G:V & B:L)
due to crossing over. Non – parental type is 17%. So the percentage of crossing over is equivalents
to 17%. The distance between the two genes (G-L) is equivalent to the percentage of crossing over
or percentage of non parental combination. So the distance between the gene G & L is equivalent
to 17 morgan units.

Percentage of non parental combination = 17%

So the percentage of crossing over = 17
So the distance between the Gene G & L = 17 map unit

In another experiment the F1 female grey red is test crossed with black cinnabar. The

experiment shows 9% non parental combination individuals. So the distance between the Gene G
& Cn is equavalent to 9 map unit.

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 In the same way the F1 female red long is test crossed with cinnabar vestigial. The

experiment shows 9.5% non-parental combination individuals. So the distance between the gene
Cn is equivalent to 9.5 map unit.
According to the first experiment the distance between G & L is equivalent to 17 map
unit. But the second and third experiment show 18.5 map units between the two genes. To find out
the actual reason for this difference in the distance, conduct a 3 point cross.
Three Point cross
In the three point cross all the three pairs of genes are considered in the experiment. The
F1 hybrid female is test crossed. They produce 8 different types of individuals. Out of 8 types, two

types are parental. Remaining six are non-parental.

Male Female
Parent : Grey Red Long Black cinnabar Vestigial
G Cn L x g cn l
Y-chromosome g cn l
Normal
F1 : G Cn L
g cn l
Back cross : Female Male
Normal Recessive
G Cn L x g cn l
g cn l Y – chromosome

Chromosome Maps of Drosophila

The chromosome maps of Drosophila include four linkage groups corresponding to four
chromosome pairs. The genes present in the X chromosome constitute the first linkage group,

those present in 2nd and 3rd chromosome constitute 2nd and 3rd linkage groups and those on the
fourth chromosome form fourth linkage group. The fourth linkage group is the smallest of all.

Chromosome maps of maize

Chromosome maps of maize have been drawn by R.A. Emerson. As there are 10 pairs of
chromosome 10 chromosome maps are seen.

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THREE POINT TEST CROSS
In a three point test cross, eight different phenotypic classes are obtained. These eight
classes are identified in two different ways, viz., (1) by phenotypic frequencies, and (2) by
alteration of gene sequence in the genotype as a result of single crossing over or double crossing
over between three linked genes. Parental types have the maximum phenotypic frequencies, double
crossovers have the lowest phenotypic frequencies, and the single crossovers have phenotypic
frequencies between these two classes. Suppose, ABC/abc are three linked genes located on two
different chromosomes in F1 of a cross between AABBCC and aabbcc parents.

1. Single crossover between A and B will alter the position of two genes, viz., B and C
2. Single crossover between B and C will alter the position of only one gene, i.e., C and,
3. Double crossover between A and C will alter the position of only middle gene, i.e., B

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Factors affecting the mapping
Chromosome map can be constructed only with the help of crossing over percentage. The
crossing over percentage is highly modified by the interference and coincidence.

INTERFERENCE
The term interference was coined by Muller which refers to the tendency of one crossover to
reduce the chance of another crossover in its adjacent region. Interference is affected by gene
distance on the chromosome. Lesser the gene distance greater is the interference and vice versa.
Generally, it is observed that crossing over in one region of chromosome may check the crossing
over in the second region.
Sometimes, presence of recombination in one region enhances the chance of recombination
in another adjacent region. This is termed as negative interference. This type of situation has

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been observed in some lower organisms, viz., Aspergillus and bacteriophages. Coefficient of
interference is estimated as follows :
Coefficient of interference (%) = 1-Coefficient of coincidence x 100
Positive and negative interference differ from one another in three main aspects .

Differences between positive and negative interference

Positive Interference Negative Interference

1. One crossover reduces the chance of One crossover enhances the chance of another
another crossover in the adjacent region. crossover in the adjacent region.
2. Observed in both eukaryotes and Found in some lower organisms like
prokaryotes Aspergillus and bacteriophages.
3. In this case coefficient of coincidence is In this case coefficient of coincidence is
less than one. always more than one.

COINCIDENCE
This term was also coined by Muller to explain strength or degree of interference. The
coefficient of coincidence is the percentage ratio of observed double crossovers to the expected
double crossovers. The greater the coincidence, lesser will be the interference and vice versa. Thus,
Observed double crossovers
Coefficient of coincidence (%)= ----------------------------------------- 100
Expected double crossovers

Coefficient of coincidence is a measure of the intensity of interference, because it has negative

association with interference. The value of the coefficient of coincidence is less than 1 for
positive interference, greater than 1 for negative interference, 1 for absence of interference and
zero for complete or absolute interference.

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Lecture 24. Sex determination: Autosomes and sex chromosomes - chromosomal theory of
sex determination - different types – sex determination in human, fowl, butterfly,
grasshopper, honey bee, fumea; Genic balance theory of Bridges, quantitative theory,
hormonal theory, barr bodies, metabolic differentiation theory; Gynandromorphs – sex
reversal in chicken

Sex differentiation in living organisms into male and female causes morphological, physiological
and behavioral differentiation between the two sexes and this phenomenon is called sexual
dimorphism. In a large number of species of animals and a small number of species of plants, eggs
and sperms are produced by different individuals, viz., females and males respectively.

MONOECIUOS
• Many plants and some animals (earthworms and hydra) have both male and female sex
organs in the same individual and produce both male and female gametes (sperm and egg,
respectively). These organisms are monoecious. (maize)
DIOECIOUS
• organisms come in two sexes, ie., male and female in separate individuals, and each
individual will produce only one type of gamete (papaya, palm)
HERMOPHRODITES
• Plants/flowers where both the male and female organs occur together (paddy, Hibsicus)
GYNANDROMORPHS
• In Drosophila some individuals show male characteristics in a part of their body, while
their remaining parts show the female phenotype, such individuals are known as
gynandromorphs. This is mostly caused by irregularities in cell division during embryonic
development

A sex-determination system is a biological system that determines the development of sexual

characteristics in an organism.
• Most sexual organisms have two sexes. In many cases, sex determination is genetic:

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• males and females have different alleles or even different genes that specify their sexual
morphology.
• In animals, this is often accompanied by chromosomal differences. In other cases, sex is
determined by environmental variables (such as temperature) or social variables (the size
of an organism relative to other members of its population

How sex is determined in animals?

sex is determination by
1. sex chromsome
2. Genic balance
3. environment
History
• Henking (1891) first observed the X chromosome in male insects
• McClung also observed extra chromosomes in in males of grasshopper
• Wilson (1909) – named X chromosome
• Stevens (1909) – named Y chrosome

SEX CHROMOSOME (ALLOSOME)

Heteromorphic chromosomes whose distribution in a zygote determines the sex of the organism
(usually designated as X or Y)
AUTOSOME
chromosome other than a sex chromosome and there is no difference in autosomes in male and
female
Sex chromosomes
• X and Y are the two sex chromosomes named in animals and plants
• The X chromosome is one of the two sex-determining chromosomes in many animals
Species,
• The X chromosome was named for its unique properties by early researchers,
• Y chromosome was named so as it it was discovered later and comes next to X in the
alphabet

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• Usually X chromosomes are larger than Y with some exception in plants
• X chromosmes carry more genes than Y

Barr body is the inactive X chromosome in a female cell ie., out of two X chromsomes of
human beings one of the X chromosome is inactivated and called BARR Body
• rendered inactive in a process called Lyonization ( in cells with multiple X
chromosomes, all but one is inactivated during mammalian embryogenesis )

SEX DETERMINATION BY SEX CHROMOSOME – Chromosomal mechanisms of sex

determination
• The different mechanisms of chromosomal sex determination may be grouped into five
classes
• 1. XX female, XY male
• 2. XY female, XX male
• 3. XX female , X- male
• 4. X- female, XX female
• 5 diploid female and haploid male
usually most of the animals have one pair of sex chromosomes. However, in Ascaris incurva, there
are 8X chromosomes in females and one Y chromosome in males; In case of Blaps polychresta
the males have 12 X and 6 Y chromosomes while the females have 12 pairs of X chromosomes
alone

1. XX-XY system
• XX female - XY male
• The XY sex-determination system is a well-known sex determination system.
• The XY sex determination system was first described independently by Nettie Stevens
and Edmund Beecher Wilson in 1905
• It is found in humans, most other mammals, some insects (Drosophila) and some plants
(Ginkgo).
• In the XY sex-determination system, females have two of the same kind of sex

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 chromosome (XX), and are called the homogametic sex.
• Males have two distinct sex chromosomes (XY), and are called the heterogametic sex

2. XY-XX sex-determination system -found in birds and some insects and other organisms.
• Females have two different kinds of chromosomes (XY), and heterogametic
• Males have two of the same kind of chromosomes (XX), Homogametic

3. XX - X0 sex-determination system
• in this system only one type of sex chromosome viz., the X chromsome is present
• in this system in one sex two X chromosome are present in another sex there is only one
sex chromosome, X_(XO)
• XX female- X0 male
• found in grasshoppers, crickets, cockroaches, and some other insects
• In this system, there is only one sex chromosome, referred to as X. Males only have
one X chromosome (X0), while females have two (XX).
• The zero (sometimes, the letter O) signifies the lack of a second X chromosome.
• Female gametes always contain an X chromosome, so the sex of the animals' offspring
is decided by the male.
• Its sperm normally contain either one X chromosome or no sex chromosomes at all.
• In a variant of this system, certain animals are hermaphroditic with two sex
chromosomes (XX) and male with only one (X0). The model organism Caenorhabditis
elegans — a nematode frequently used in biological research — is one such organism.

4. XO-XX system
• In fumea this kind of sex determination is found
• XO- females
• XX- males

5. Haplodiploid sex-determination system

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• The Haplodiploid sex-determination system determines the sex of the offspring of
many Hymenopterans (bees, ants, and wasps), and coleopterans (bark beetles).
• In honeybees
females – diploid
males – haploid

II. SEX DETERMINATION BY GENIC BALANCE THEORY

• In Drosophila, the greater the number of X chromosomes relative to the autosomes, the more
likely the individual will be female
Sex balance theory or genic balance theory states that the X chromosome determines
the sex of the individual and that sex is a dosage phenomena,
• where the ratio of the amount of the X relative to the autosomes determines the sex.
• In addition, environmental effects can influence the development of the intersex flies.
• sex index = number of X chromosomes

number of autosomes
• If the ratio is =1 - females
= >1.0 - super females
=0.5 - male
= <0.5 – super males
= 0.5 – 1.0 - intersex

Genic Balance theory of sex determination (Bridges 1921)

A number of lines of evidence indicate that even in dioecious species, all individuals have
genes for both sexes. To quote Bridges, `Both sexes are due to the simultaneous action of two
opposed sets of genes, one set tending to produce the characters called female and the other to
produce the characters called male. `Which sex actually develops is decided by the balance, i.e.,
by the preponderance of the female-determining or of the male-determining genes. The sex
chromosomes are merely vehicles of genes which help in tilting the balance in one direction or
another.

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 Support for the balance theory of sex determination comes from the work of
Bridges(1921) on Drosophila. Bridges observed some females of Drosophila melanogaster with
three X chromosomes and three sets of autosomes (i.e, triploids). When he crossed them with
normal (diploid) males, he found that some of the progeny had one or more chromosomes less or
more than the normal flies(i.e., aneuploids). His results are given below:

X+A Y+A
2X + 2A 3X + 3A 2X + Y + SA
Female Intersex
X+A 2X + 2A X + Y + 2A
Female Male
2X + A 3X + 2A 2X + Y + 2A
Superfemale Female
X + 2A 2X + 3A X + Y + 3A
Intersex Supermale

Bridges found intersexes, superfemales and supermales among the progeny. Intersexes are
sterile individuals intermediate between females and males but are different from gynandromorphs
which are typically female in certain portions of the body and typically male in others.
Superfemales and supermales are sterile individuals which are very weak and very poor in viability.
This is shows that in Drosophla the X chromosomes carry genes that are predominantly
female-determining.
Flies with one X, one Y and two sets of autosomes are normal males but flies with one X,
one Y and three sets of autosomes are supermales.
1X + 1Y + 2A Male
1X + 1Y + 3A Supermale

This shows that the autosomes carry genes that are predominantly male-determining.

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 That individuals with two X, one Y and two sets of autosomes are female in spite of the
presence of the Y chromosome shows that the Y chromosome plays no positive role in sex
determination. That the Y chromosome does not determine maleness in also shown by the fact that
flies with one X chromosome and two sets of autosomes (i.e., XO flies) are males in spite of the
absence of the Y chromosome. They are, however, sterile showing thereby that the Y chromosome
contains male fertility genes necessary for the production of a fertile male.

Bridges interpreted these results as follows:

Sex in Drosophila melanogaster is determined by the X chromosomes as well as by the
autosomes, the ratio of the number of X chromosomes to the number of sets of autosomes being
the deciding factor
Phenotype chromosomal complement number of X chromosomes/
number of autosomal sets
normal female XX + 2N autosomes 1.00
normal male XY + 2N autosomes 0.50
superfemale XXX + 2Nautosomes 1.50
superamale X + 3N autosomes 0.33
intersex (sterile) XX + 3N autosomes 0.67

III. Environment –sex determination

• Many other exotic sex-determination systems exist. In some species of reptiles, including
alligators, some turtles, and the tuatara, sex is determined by the temperature
at which the egg is incubated.
• some hormones produced by the proboscis of female Bonelia induces larvae to
differentiate into males
• in Dinophilus(sea worm) , animals from larger sized eggs are females and from smaller
size are males
• Other species, such as some snails, practice sex change: adults start out male, then
become female.

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• In tropical clown fish, the dominant individual in a group becomes female while the
other ones are male.
• In some arthropods, sex is determined by infection. Bacteria of the genus Wolbachia
alter their sexuality;
• some species consist entirely of ZZ individuals, with sex determined by the presence of
Wolbachia

Lecture 25. Sex linked inheritance – cris cross inheritance – reciprocal difference;
holandric genes; sex influenced and sex limited inheritance.

SEX LINKAGE- location of gene on X chromosome or sex chromosome

Morgan crossed a red eyed female with a white eyed male and found that all F1 flies of
both sexes were red eyed. In F2, 3 red and 1 white eyed. So, it is due to an allelic Pair of genes of
which red is dominant.

Red female X White male

F1 Red female Red male

F2 Red female Red female Red male White male

A reciprocal cross was made between white eyed female and red eyed male. It
was found that among the F1 off spring, all the females were red eyed and all the males were white
eyed. The results were quite unexpected firstly, because the phenotypes of F1 females and male
were different.
White female X Red female

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 F1 Red female White male

F2 White female Red female White male Red male

The different results from the reciprocal crosses could be explained only on the assumption
that the gene for eye colour is located on ‘X’ chromosomes and that ‘Y” chromosome has no gene
for colour of the eyes. A white eyed female crossed with a red-eyed male produces red eyed females
and whtie eyed males, this method of inheritance, is often referred to as ‘criss-cross inheritance’.
The F2 consisted of red eyed and white eyed individuals in equal numbers in both sexes.
Morgan concluded that the gene for eye colour is located on the X chromosome and that
the Y chromosome carries no gene for eye colour.

Sutton and Boveri hypothesised that genes are borne on chromosome (i.e, the chromosome
theory of heredity) but it was Thomas Hunt Morgan (1910) who first associated a particular gene
(i.e., the gene for eye colour) with a particular chromosome (i.e., X chromosome) visible in
microscopic preparations and showed that the gene for eye colour follows exactly the transmission
of the X chromosome.

Criss cross inheritance:- A sex linked gene passes from male to female then back to male. The
gene for eye colour is located on ‘X’ chromosome, it is called ‘X’ linked gene. This pattern of
inheritance is called ‘Sex linkage’.

There are genes located on ‘Y’ chromosomes and its allels absent in X chromosome. Such
gene are called ‘Y linked’ or Holandric genes. The gene responsible for hypertrichosis causing
hairy prina (ear lobes) in human beings is a Y linked gene. There are certain homologons regions
on X and Y chromosomes in which both the allels of a gene may be present as in the case of bobbed
bristles (b) and its allele (b+) for normal bristle. Such genes are present both in X and

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Y chromosomes are called XY linked genes. Eg. Genes for colour blindness, Xerodermia
pigmentosum (pigmentation on the skin), Retinitis pigmentosa (pigmentation on the eye retina),
Nephritis etc., in human beings are XY-linked.

Sex-influenced characters: are characters which may be expressed differently in the two
sexes even when their genotypes are identical. The mere influence of the sex of the individual may
be sufficient to alter the phenotypic expression of a gene. The most common expression of sex
influence is that dominance is reversed between the sexes. Genes determining sex influenced
characters are borne on autosomes.
A typical example of a sex-influenced character is the presence of horns in sheep. Both
sexes of Dorset sheep are always horned while both sexes of Suffolk sheep are always hornless. If
Dorset and Suffolk are crossed, the F1 females are hornless while the F1 males are horned. One

interbreeding the F1 sheep, an F2 is obtained in which the females show a ratio of 3 hornless

: 1 horned and the males show a ratio of 3 horned : 1 hornless. Presence of horns may therefore be
said to be recessive character in females but a dominant character in males.
P Horned female X Hornless male
HH Hh
F1 Hh
Females, hornless
Males, horned

F2 1 HH : 2Hh : 1hh

Female Horned Hornless Hornless

Male Horned Horned Hornless

Reciprocal crosses show no differences because the gene is carried by the autosome.
Baldness in human beings is a sex-influenced character which is recessive in females and dominant
in males.
Sex-limited character

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 Sex-limited inheritance is an extreme type of sex influence in which a particular phenotype
can be expressed only in one sex. As genes for sex-limited characters are borne on autosomes, all
genotypes should occur with identical frequencies in both sexes but the physiological differences
between the sexes are such that certain genotypes can be expressed only in one sex. Unlike sex
influenced characters in which gene is dominant in one sex and recessive in the other, sex-limited
characters are controlled by genes which have no visible influence at all in one sex-either as a
homozygote or as a heterozygote.
In domestic poultry, cock-feathering is a character limited to the male sex. Hen-feathering
is due to a dominant gene H and cock-feathering is due to its recessive allele h, but females with
genotype hh are hen-feathered. The genotypes and their corresponding phenotypes are as follows:

Genotype Phenotype
Female Male
HH Hen-feathered Hen-feathered
Hh Hen-feathered Hen-feathered
Hh Hen-feathered Cock-feathered

Removal of the ovaries in hens with genotype hh results in cock-feathering. This indicates
that the female sex hormone inhibits the production of cock-feathering in hens with genotype hh.
Sex reversal
In several species of plants that ar normally bisexual, suppression of the male or female
structuers has been observed in nature. The androecium getting converted into petals in ornamental
plants or carpels as in carrot and cabbage or pistils as in maize, papaya and primrose has been
observed. When the stamens get converted into rudimentary organs without the pollen sac and
pollen, they are called staminodes and a similar convertion of the pistil into ononfunctional
rudimentary organ is called the pistillode. The phenomenon in which there is

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suppression of one sex at the expense of the other is called sex reversal. The sex reversals are
mostly due to physiological and biochemical alterations involving sex hormones.
In maize, rarely it is observed that the male inflorescence called tassel bears seeds due to
sex reversal. The recessive gene `ba' is responsible for barren plants and another recessive gene
`ts' is responsible for tassel seed. Sex reversal in maize is due to the genetic constitution of the
plants.

Lecture 26. Sex determination in plants – Melandrium, papaya, maize.

Lecture 27. Cytoplasmic inheritance and maternal effects – features of cytoplasmic

inheritance, chloroplast, mitochondrial - plastid colour in Mirabilis jalapa - iojap gene of
maize, cytoplasmic male sterility in rice, kappa particles of paramecium - plasmid and
episomic inheritance.

Plastids and mitochondria, localized in the cytoplasm have been shown to be

responsible for the extranuclear transmission of inherent qualities. Like the nuclear genes,
they are capable of specific self-duplication. They are transmitted from generation to generation.
They mutate and the mutant particles reproduce their own kind just as do mutated genes.

The totality of heredity transmitted through the cytoplasm is referred to as plasmon, and
all cytoplasmic particles which manifest genic properties, viz., self-duplication, specificity and
mutability are called plasmagenes.

Cytoplasmic inheritance: The transmission of hereditary characters by the cytoplasm.

Inheritance via genes found in cytoplasmic organelles.
A few traits are controlled by genes located in cell organelles in the cytoplasm

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 i.e. cytoplasmic genes,
• These genes are exceptions to the chromosome theory of inheritance
• The inheritance of genes from the female parent that are not in the nucleus but in
organelles such as mitochondria that are found in the cytoplasm.

The cytoplasmic inheritance is known as extrachromosomal inheritance

Extra-chromosomal inheritance - controlled by non- nuclear genomes
• The inheritance of genes from the female parent that are not in the nucleus but in
organelles such as mitochondria that are found in the cytoplasm.
• This type of inheritance is not controlled by Mendel's laws.
• Because the cytoplasm is usually contributed entirely by one parent,the characteristics
encoded by these genes are usually inherited from only that parent.

Extrachromosomal inheritance

Cytoplasmic (maternal) maternal effect

Inheritance

plastids mitochondria symbionts like bacteria virus

ORGANELLE DNA
Genes located outside of the nucleus (i.e., not on the major chromosomes)
• Mitochondrial genomes (mt DNA)
• Chloroplast genomes (cp DNA)
• extranuclear plasmids and other factors

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• mtDNA and cpDNA are almost always uniparentially inherited, with only one sex (typically
the female) transmitting the genomes to their offspring.
• The cytoplasm contributed by the female contain several components

Mitochondrial inheritance in petite mutants of Yeast

• Colonies smaller in size than the normal colonies are occasionally observed in the yeast
(Saccharomyces cerevisiae) when grown on solid medium. These small colonies are called
petite mutants.
• Petites have slow growth as they grow by gulcose fermentation anaerobically and hence
inefficient in metabolism as compared to the normal colonies with aerobic metabolism.
• Among the petite mutants, there are segregational petites that exhibit Mendelian
segregation when crossed with wild type and the petite is recessive to wild type.
• Another class of petite mutant is called neutral petite, which on crossing with wild type
produces only wild type colonies and is thus uniparental in inheritance. The reason for such
a differential behaviour seems to be that majority of neutral petites lack most or all
mitochondrial DNA responsible for oxidative respiration.
• The petite phenotype is the result of large deletion in mitochondrial DNA. Thus the
mitochondria is found to be responsible for inheritance in respiration defective mutants in
yeast.
Inheritance of Kappa particles in Paramecium
• The presence of symbionts such as bacteria and viruses living in the cytoplasm was found
to be responsible for a variety of cytoplasmically transmitted characters in eukaryotes.
• In Paramecium aurelia, the presence of bacterial inclusions known as kappa particles
produce killer phenotype and its maintenance is dependent on a dominant nuclear gene K.
the killer strain releases a substance lethal to many other strains of the protozoan.
• When conjugation occurs between the killer (KK) strain and the sensitive (kk) strain and
if no cytoplasmic exchange takes place between them, the resulting progeny have killer
and sensitive types in equal proportion eventhough the genetic constitution of both types
is Kk. In the further progeny resulting form autogamy, the killer produces killer and
sensitive types while the sensitive produces only sensitive types.

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 • On the other hand, when cytoplasmic exchange occurs between the killer (KK) and
sensitive (kk) strains during conjugation, all the resulting progeny are killer (Kk) and no
autogamy, each killer produces killer and sensitive types in equal number.
• They are diagrammatically presented below:
• When cytoplasmic exchange takes place between the killer and sensitive strains, the kappa
particles in the cytoplasm of the killer strain mixes with the cytoplasm of the sensitive
strain and in the presence of the dominant allele K in the heterozygote Kk, the kappa
particles cause the killer phenotype. A
• fter autogamy, half of the progeny have cytoplasm with kapppa particles along with the
dominant gene K and hence turn to be killers while the other half without both kappa
particles and dominant gene turn to be sensitive. Thus an interaction between nuclear gene
and cytoplasmic inclusion has been found to be responsible for inheritance of killer trait in
Paramecium.

Maternal effect - Shell coiling in Snail

• There are instances in which the phenotype of the offspring is neither determined by the
genotype of both its parents nor by the cytoploasmic or extra nuclear factors.
• In the snail Limnaea peregra, the direction of coiling of the shell is either dextral (right
ward coiling) or sinistral (leftward coiling). When a dextral female is crossed to a sinistral
male, the F1 is dextral.

• On the other hand, when a sinistral female is crossed to a dextral male, the F1 is sinistral.

All the F2 offspring produced by self fertilization (as the snail is hermaphroditic) are

dextral in both the direct and reciprocal crosses. In this case, though, the F1 behaviour

resembles maternal inheritance, the behaviour F2 does not conform to it. When the F3

offspring obtained by self fetiliszation of the F2 were examined, they segregate in the

proportion of 3 dextral to one sinistral.

• Sinistrality is governed by the recessive allele s and dextrality by its dominant type allele
+. Thus the parents in the direct cross are dextral female (+/+) and sinistral male (s/s) and

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 vice versa in the reciprocal cross. Yet the heterozygous F1 (s/+) is dextral in the direct

cross, but sinistral in the reciprocal cross. This is due to the maternal effect, which means
that the genotype of the mother determines the phenotype of the offspring.
• Similarly the F2 offspring possess the genotypes +/+, +/s, +/s and s/s. The F3 offspring of

s/s F2 are sinistral while all other F3 offspring are dextral. It has been proved that the

orientation viz., dextrality or sinistrality is predetermined in the egg cytoplasm as a result

of the mother's genotype, the mitotic spindles during the second cleavage division in the
zygote determining the orientation. This is a case of maternal effect distinctly different
from maternal inheritance.
Inheritance of plastids in Mirabilis
In a wide variety of plants, there occurs a type of leaf variegation in which the normal green
leaves have pale green or white patches. These patches may be small or may extend throughout
the leaves.
• The inheritance of plastids in the Four O' clock plant, Mirabilis jalappa was first described
by Correns (1908). In Mirabilis jalappa, some of the branches may have normal green
leaves, while in the same plant, some other branches may have only pale green or white
leaves and still others may have variegated leaves.
• Flowers on branches with normal green leaves produce seeds that grow into plants with
normal green leaves irrespective of whether they are pollinated by pollen from branches
with normal green, variegated or pale green leaves.
• Flowers on pale green branches produce seeds that grow into plants with only pale green
leaves regardless of the pollen parent.
• Flowers on variegated branches yield offspring of three kinds, viz., green, variegated and
pale green in variable proportions irrespective of whether they are pollinated by pollen
from branches with normal green, variegated or pale green leaves.
Progeny of a variegated four O' clock plant
Type of branch from which Type of branch from which Type of leaf in the progeny
flowers are chosen for pollen was obtained grown from seed
pollination

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 Green Green Only green
Variegated Only green
Pale green Only green
Variegated Green Green, variegated or pole
Variegated Green, variegated or pole
Pale green Green, variegated or pale
Pale Green Green Only pale green
Variegated Only pale green
Pale green Only pale green

It is clear that variegation is determined by agencies transmitted through the female and
that it is not influenced by the type of pollen used. These agencies are the chloroplasts. They are
easily visible under the microscope as particles localized in the cytoplasm. They are capable of
self-duplication and are transmitted from generation to generation through the cytoplasm of the
egg. Seeds borne on a green branch have therefore only green plastids, seeds borne on a pale green
branch have therefore only pale green plastids and seeds borne on a variegated branch have green
or pale green or a mixture of the two types of plastids.
Variegation is thus a hereditary character determined by stable, self-duplicating,
extranuclear particles called plastids. Neither the nucleus of the female gamete not the male gamete
is involved in the control of this type of hereditary character.

Maternal inheritance by ‘iojap’ gene in maize

Rhoades (1946) identified the iojap gene (Ij) in maize located in chromosome VII
controlling plastid inheritance in the plant. The gene Ij is responsible for the normal green colour
of the plant. When normal green plants with Ij Ij are used as female and pollinated by pollen from
striped plants with ij ij, the F1 plants with Ij ij are wholly green.

Green X Striped
Ij Ij Ij ij

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 F1 : Green
Ij ij

When striped plants with ij ij are pollinated by pollen from the normal green plants with Ij
Ij, the F1 plants, all of which have the same genotype, Ij ij are of three different phenotypes, viz.,

normal green, striped and white.

Striped X Green
Ij ij Ij Ij
F1 : Green, striped or white
Ij ij

When plants with the same genotype Ij ij have different phenotypes, viz., normal green,
striped, or white, the differences can be attributed only to differences in plastids.
Cytoplasmic male sterility in maize
In several cases of cytoplasmic inheritance in plants, plastids have been shown to be the
vehicles of heredity but in several other cases, cytoplasmic particles other than plastids have been
identified as the basis for extranuclear transmission. Among these is a case of male sterility in
maize. Most or all of the pollen grains of such male sterile plants are aborted. This character is
transmitted only through the female and never by the pollen. When all of the chromosomes of the
male sterile line were replaced with chromosomes of normal plants, the line still remained male
sterile, showing thereby that male sterility is controlled by some genes in the cytoplasm. It was
later recognized that cytoplasmic male sterility in maize results from alterations in the hereditary
units in the mitochondria (mitochondrial DNA).

Lecture 28, 29, 30, 31,32 CHEMICAL

BASIS OF HEREDITY
Nature of the genetic material

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 A swiss biologist, Miescher (1869) identified a chemical compound in pus cells and salmon
sperm in the large nuclei of these cells. The chemical was named `Nuclein'. As it was found to be
acidic, it was called `Nucleic acid'. All plant and animal materials were found to contain nucleic
acid. Nucleic acid was found associated with various proteins and along with the protein, it was
called `Nucleoprotein'.
There are two kinds of proteins associated with nucleic acid and they are prolamine and
histone. Prolamine consists mostly of linked groups of amino acids arginine. Histones are
relatively complex in nature. Because of this complexity, proteins were originally thought as the
genetic material. Proteins have long chemical chains consisting of many amino acids and they were
considered to be capable of carrying many complex messages that cause variation in the biological
material.
Considering the proportion of different constituents of cell, nucleic acid was found to be
constant in volume in all the cells as compared to other cellular contents and hence it was inferred
to be the hereditary material. There are two types of nucleic acid, the deoxyribonucleic acid
(DNA) and the ribonucleic acid (RNA). By staining nucleic acid, Feulgen (1924) found that the
DNA was localized in the nucleus, while the RNA was found to occur outside the nucleus in the
cytoplasm.
The experiments of Griffith (1928) with the pneumonia bacterium and the interpretation
of results by Avery, MacLeod and McCarty (1944) confirmed the DNA as the hereditary
material.

Bacterial Transformation
Giffith (1928) worked on the pneumonia causing spherical shaped bacterium, Diplococcus
pneumoniae. Some of the strains of this bacterium have a smooth polysaccharide capsule which
causes the disease and hence called virulent S strain. A mutant strain has no capsule and is avirulent
or nonpathogenic and is called R strain. In agar medium, the virulent (S) strain produces smooth
surfaced colonies, while the avirulent ® strain produces rough surfaced colonies. There are several
types of these two strains, S I, S II, S III, R I, R II, R III etc. that

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differ in the type of antigen they produce. The kind of antigen produced in genetically determined.
The S type sometimes mutates to R type but not in the reverse.
Griffith injected the laboratory mice live R II bacteria and the mice did not get pneumonia
as R II is a virulent. When injected with virulent S III, the mice suffered of pneumonia and died.

When S III bacterial were heat killed at 65oC and then injected into the mice, they did not suffer
of the disease and lived. Later, the heat killed S III strain and live avirulent R II strain were mixed
and injected into the mice. Contrary to expectations, the mice suffered of pneumonia and died. On
analysing the blood sample of the affected mice, live S III and live R II bacteria were found in it.
This could not be possible due to the mutation of the avirulent R II to virulent types. Evidently,
some heat-stable component present in the heat killed, and hence, dead S III strain could have
conferred the virulent nature to the live R II strain. Griffith designated this as the `transforming
principle' that transformed the hereditary property of avirulent R II to virulent S III. This
phenomenon is called `Griffith effect' or `Bacterial transformation'.
Griffith did not understand the cause of bacterial transformation. Avery, MacLeod and
McCarty (1944) tested a fraction of the heat killed S III bacterial for the transforming ability. They
removed proteins, lipids, polysaccharides and ribonucleic acid from S III extract by a variety of
chemical and enzymatic methods without diminishing its ability to transfer R II into S III strain.
They found that a cell-free and highly purified DNA extract of S III bacterial could bring about
transformation of R II into S III and concluded that DNA is the transforming principle and hence
the genetic material in bacteria.
Later studies on other bacteria such as Haemophilus influenzae, Bacillus subtilis,
Escherichia coli, Shigella paradysenteriae and others revealed that they also undergo
transformation.
Transformation is the process of adding a foreign DNA fragment from a donor genome
into the genome of a recipient cell. The donor fragment passes through the cell membrane of the
recipient cell of the same or different species and becomes incorporated into the genome of the
recipient cell through recombination.
DNA as the genetic material in Viruses

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 Hershey and Chase (1952) provided direct proof that DNA is the genetic material in
certain bacterial viruses.
Bacteriophage is a virus that infects or feeds on certain specific bacteria. T2 bacteriophage

that infects the colon bacteria, Escherichia coli was involved in the studies.
Bacteriophage is eletron microscopic. It has a head and a tail. Inside the head, there is a
long chain of DNA molecule. The phage attaches itself by its tail to the bacterial and injects the
DNA into the bacillus. It dictates the cell to produce many copies of the viral DNA.
Bacteriophages are used in many finer analyses of the genetic material since they are
haploid organisms and there is no hiding of mutant effect. As there is no differential sex, there is
no need for two different individuals to unite for reproduction. They multiply enormously and have
a short life span. Recombination's and mutations, even if in a very low frequency, could be
recognised with relative ease. When a population is raised from a single phage all the descendents
will be identical. But occasionally, through errors in copying of genetic material, rare mutants
appear and such mutants are called `copy error'.
In a chemically defined cultural medium, known quantities of radioactive isotopes of

phosphorus P32 and sulphur S35 were added. Escherichia coli were grown in the medium and the
labelled E. coli cells were used as hosts for unlabelled T2 bacteriophage. The virus progeny that

multiplied inside the bacteria could be traced in the culture medium on lysis (cell wall breakage)
of the bacteria.

The viral DNA was labelled with P32 and the viral capsid (protein coat) with S35, since
DNA contained P and the capsid protein contained S. then the labelled viruses were allowed to
infect unlabelled E. coli and get multiplied. Later the viruses were separated from the bacterial

host cell by agitation and the content of P32 and S35 of the virus and bacteria was assessed. P32
could be traced in the infected bacterial cells. Hershey and Chase inferred that DNA of the virus
entered the bacterium and played a role in viral multiplication, whereas the protein of the virus did
not play any role in the intef cellular replication of the virus. Thus it was established that the genetic
material of the virus was DNA.
RNA as the genetic material in some viruses

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 Ribonucleic acid was found to be the genetic material in Tobacco mosaic virus, Turnip
yellow mosaic virus, Poliomyelitis, Foot and Mouth virus, Influenza virus, Reovirus, Rous
sarcoma virus and some of the bacteriophages such as MS 2.

Properties of the genetic material

The biological information properties of the genetic material enable the capability to
provide, from one generation to another, the information that gives each species its ability to
reproduce its own kind accurately. These include structural gene properties and regulatory gene
properties.
The genetic material must be capable of specifying the structure of other molecules. The
genetic material should be capable of accurately coding for a large number of different polypeptide
amino acid sequences during protein synthesis. The genetic material should regulate the
programmed of embryological development and organ differentiation for a given species, which
differ from one species to another. Thus the genetic material should be able to regulate gene action
and function.
The genetic material has the replicative properties. During mitosis, each daughter cell
receives a copy of all the genetic information from the parent cell. During meiosis, each gamete
receives a copy of half the genetic material.
Mutational properties of the genetic material provide a fundamental source of variation
essential for evolution of species.

Chemical composition of DNA

DNA is a complex macromolecular or polymeric chemical compound which contains four
kinds of monomers (small building blocks) called Deoxyribonucleotides. Each
deoxyribonucleotide is made up of 1 ) a phosphoric acid molecule, biologically called phosphate,
discovered by Levene (1910), 2) a pentose sugar called 2 -deoxyribose, also discovered by Levene
(1910) and 3) four major kinds of nitrogen bases, two heterocyclic and two ringed purines, adenine
(A) and guanine (G) and two one ringed pyrimidines, cytosine (C) and thymine (T), discovered by
Fischer (1880).

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 A nucleotide results from covalent bonding of a phosphate and a nitrogen base to the
pentose. That part of each nucletide which contains a nitrogen base and deoxyribose is called
Deoxyribonucleoside.
The four kinds of deoxyribonucleosides and deoxyribonucleo tides are as follows:
Nucleosides and nucleotides of DNA
Nitrogen base N base + Deoxyribose = Deoxyribo nucleoside + Nucleotide
Deoxyribo nucleoside Phosphoric acid =
Deoxyribo nucleotide
Adenine Deoxyadenosine Deoxyadenylic acid D e o x y- a d e n o s i n e
mono-phosphate
(dAMP)
Guanine Deoxyguanosine Deoxy-guanylic acid D eo x y- gu a n o s i n e
nono-phosphate
(dGMP)
Thymine Deoxythymidine Deoxythymidylic acid Deoxythymidinemon
o phosphate (dTMP)
Cytosine Deoxycytidine Deoxycytidylic acid Deoxycytidine
mono-phosphate
(dCMP)

Two organic chemists, Levene of the Rockefeller Institute and Todd of Cambridge (1910)
demonstrated that the components of DNA were joined together to form a long chain of alternating
deoxyribose and phosphoric acid units with side chains of the nitrogen bases.
Double helical model of DNA
Based on the findings of Chargaff (1950) that the total amount of purines equalled the
total amount of pyrimidines (A + G = T + C), that the amount of adenine equalled the amount of
thymine (A = T) and the amount of guanine equalled the amount of cytosine (G = C) and, that the
ratio between total purines and total pyrimidines was always not far from one, (A + G) : (T + C)
= 1, as well as the crystallographic evidences and X-ray differentiation photographs (Astbury,

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1947, Wilkins and Franklin, 1953), the double helical model of DNA was constructed by
Watson, an American biologist and Crick, a British physicist in 1953.
The DNA molecule was conceived as a two stranded structure coiled like a rope, and hence
called plectonemic, so that if the ends are permitted to revolve freely, the complementary strands
could easily separate. The coil was proposed to be helical and conceived to resemble a circular
staircase, maintaining the same diameter throughout and having a constant width between steps.
The steps are connected on either side by a railing.

The helix has a diameter of 20 Ao and makes a complete turn at every 34 Ao along its

length. The distance between nucleotides is 3.4 A o. Each complete turn has a stack of 10
nucleotides. The helix contains two polynucleotide chains or two stacks of 10 nucleotides each per
turn.
Each complementary strand is only half the circular staircase, either side consisting of
approximately half the width of the step. Each half step is connected by a railing or backbone. The
railing consists of phosphate - sugar linkages which are repeated without change. The half step of
one strand extends to meet the half step of the complementary strand. Each half step has either a
purine or pyrimidine base. Each step consisting of two half steps is together called base pair.
The fit between the bases is determined by hydrogen bonding. The bonding involves the

ability of the H atom with positive charge (H+) to be placed between an O atom with weak negative

charge (O-) and a N atom with a light negative charges (N-) from opposite strands. Adenine pairs
with thymine with two H bonds (A = T) and guanine with cytosine with three H bonds (G = C).
These N bases are connected to each other by deoxyribose and phosphoric acid.
Hydrogen bonds are generally weaker than other chemical bonds. But there are several of
them, two between A and T (A = T) and three between G and C (G = C) that give rigidity and
stability to the molecules.
The position of purines, pyrimidines, deoxyribose and phosphoric acid in DNA is presented
in Fig. 17-4.
Chemical composition of RNA

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 In RNA, there are four nitrogen bases, the purines consisting of adenine and guanine and
the pyrimidines consisting of cytosine and uracil. Thymine in DNA is replaced by uracil in RNA.
For uracil, the ribonucleoside is uridine and ribonucleotide is uridylic acid or uridine mono
phosphate (UMP). The ribonucleosides for adenine, guanine and cytosine are adenosine,
guanosine and cytidine respectively and the ribonucleotides are adenylic acid (or adenosine
monophosphate - AMP), guanylic acid (or guanosine monophosphate - GMP) and cytidylic acid
(or cytidine mono phosphate -CMP) respectively.
The structure of ribose sugar (as different from deoxyribose in DNA) and uracil (U) are as
follows:
The methyl group at position 5 in the thymine is replaced by a H atom in uracil.
RNA is made up of many ribonucleotides. The ribose sugar and phosphoric acid remain linked by
phosphodiester bonds.
Organisms which have only RNA employ their RNA in their genetic mechanism. Such
RNA is called genetic RNA. Organisms having DNA along with RNA, use the RNA for carrying
the orders of DNA and such RNA is called nongenetic RNA. Ribosomal RNA (rRNA), transfer
RNA (tRNA), heterogeneous RNA (htRNA) and messenger RNA (mRNA) are nongenetic
RNAs. rRNA and mRNA are single stranded, while tRNA and htRNA are doubled stranded. The
genetic RNA present in most of the viruses is single stranded while that in reterovirus is double
stranded.

24. REPLICATION OF DNA AND RNA

Replication of DNA
DNA has two important functions as the carrier of genetic information.
1) When DNA directs the synthesis of DNA itself, or in other words, when DNA
replicates, such a function is autocatalytic.
2) When DNA directs the synthesis of chemical molecules other than itself, such as
synthesis of RNA, protein etc., such a function is heterocatalytic.
The double helical model of DNA proposed by Watson and Crick provides a template
system for self-replication. Because of the specificity of base pairing, A with T and G with C, the
sequences of bases along one chain automatically determines the base sequence along the

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other. Hence each chain of the double helix can serve as a template for the synthesis of the
other.
Replication involves disruption of H bonds, followed by a rotation and separation of the
two-polynucleotide strands. Each purine and pyrimidine base of each polynucleotide strand
attracts a complementary free nucleotide available for polymerisation in the cell and holds it in
place on the paternal template chain. The free nucleotides are sewn together by formation of
phosphate di ester bonds for linking adjacent deoxyribose, thus forming a new polynucleotide
molecule. Thus two double helical molecules, identical to each other are formed.
Three theories were proposed for DNA replication. They are semiconserative theory,
conservative theory and dispersive theory.
Semiconservative theory
According to this theory, both the complementary strands in a DNA molecule separate and
each strand functions as a template or mould to prepare its replica.
Conservative theory
The strands are not supposed to separate from each other, but a new double helix appears
within the old double helical strands.
Dispersive theory
Each of the strands of the double helix breaks into pieces and these pieces duplicate. The
broken and duplicated pieces are reconstructed into two double helices consisting of strands
containing both old and new pieces.
Proof for semiconservative DNA replication - Meselon and Stahl's (1958) experiments with

Escherichia coli by labelling the DNA with heavy isotopes of N viz., N 14 and N15 and
distinguishing the density difference in a cesium chloride gradient by using sedimentation
equilibrium centrifugation and observing the band on ultra violet tube proved that DNA replication
occurs by the Semiconservative methods.
Cairns (1963), by autoradiography, studied the DNA replication of E. coli, which has a
single large duplex circular chromosome, and suggested that the chromosome replicated as an
intact ring and the model of replication was semiconservative.
A rolling circle model of DNA replication is suggested for bacteria and viruses.

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 Taylor, Woods and Hughes (1957) demonstrated that eukaryotic DNA replicates semi
conservatively in studies conducted with Vicia faba.
Some of the nucleic acid enzymes play an important role in DNA replication. They are: 1)
Nucleases that catalyse the breakdown of particular bonds leading to fragmentation of nucleic
acid. They may be exonucleases that attack nucleic acid at its terminal nucleotide only or
endonucleases that react only with those bonds that occur within the interior of the nucleotide
chain to cut in into pieces, 2) Ligases that join broken ends of two DNA chains by catalysing the
synthesis of a phosphodiester bond between 3'- hydroxyl group and 5' - phosphate group and
restore an intact DNA duplex, 3) Restriction enzymes that produce breaks only within sequences
which have two identical bases in adjacent position, 4) Swivelases that allow unwinding of one
of the two strands of DNA to allow free rotation within DNA molecule and 5) Polymerases that
are involved in the synthesis of nucleic acids by addition of bases to a growing nucleotide chain.
The speed of DNA replication was studied in vivo and in vitro by Kornberg et al. (1967)
using ø X 174 phage. The speed was 500 to 1000 nucleotides per minute in vitro and as high as
100,000 nucleotides per minute in vivo.
Replication of RNA
The genetic RNA of viruses is self-replicating. Its model of replication is called `RNA
dependent RNA synthesis'. RNA polymerase enzyme mediates in the replication mechanism
keeping the parent RNA as the template and synthesizing a complementary RNA chain.

25 & 26. PROTEIN SYNTHESIS AND GENETIC CODE CONCEPT

One gene - one enzyme hypothesis
When we consider the phenotype of an organism, we should understand that it is the result
or end product of many complicated actions and interactions within and between genes.
Phenogenesis is the mechanism by which the phenotype of an organism is produced under the
control of DNA in a given environment, which includes not only external factors such as
temperature, intensity and quality of light but also internal factors such as enzymes and hormones.
The phenotype is the result of various embryological and biochemical activities involving
enzymatic proteins. Enzymes catalyse in separating or uniting different cellular

147
molecules. A precursor is transformed into an end product through the production of many
intermediate products, each aided by an enzyme produced by a gene and this constitutes the
biosynthetic pathway.
DNA itself does not have enzymatic character and does not directly involves in the
biosynthesis pathway. The immediate gene product is a kind of RNA called messenger RNA
(mRNA) which controls the amino acids to form enzymes at the surface of the cytoplasmic
ribosomes. Thus, DNA transcribes mRNA which translates protein that ultimately produces a
phenotypic trait.
Beadle and Tatum (1941), the Nobel Laureates of 1958, working on the bread mould,
Neurospora, analysed the biochemical effects of genes by studying wild type prototrophs (strains
that grow one minimal medium) and auxotrophs (nutritional mutants that grow on supplmented
media). They found that ultra violet mutants produced defect in enzyme or loss of specific enzyme.
This concept is known as "one gene - one enzyme hypothesis".
Protein and Enzymes
Enzymes are proteins that composed of subunits called polypeptides, which can be further
broken into amino acids. The amino acids are united by peptide linkage. Though there are 35
different amino acids in biological systems, most of the biological proteins contain only 20 amino
acids.
Amino acids consist of four sub units viz., 1) a carboxyl group with a potential negative
charge, 2) an amino group with a potential positive charge, 3) a hydrogen (H) subunit and 4) an R
subunit which differentiates the amino acids.
Proteins have several structural levels
1. primary structure consists of the linear sequence of amino acids in a polypeptide chain.
2. Secondary structure consists of section of primary poly peptide chain twisted or coiled
into a helix.
3. Tertiary structure consists of very long spiral chains compressed in a globular form with
extensive folding over and bending of helices.
4. Quarternary structure consists of two or more independent polypeptide chains linked
together by interchain bonds to produce complex structure.
One gene - one polypeptide concept

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 Investigations by Ingram (1957) brought to light that each gene controls the production of
a single polypeptide chain of protein molecule. The studies were on hamoglobin pigment of the
red blood corpuscles that consist of four polypeptide chains viz., , , ,  varying in number and
arrangement of amino acids and four iron containing ring compounds (heme). Hemoglobin of adult

and foetus and abnormal haemoglobins Hb3 and Hb3 were studied electrophoretically to
understand the control of polypeptide by gene.
Crick (1958) proposed that the sequence of nucleotides in DNA and of amino acids in
proteins is colinear. This means that there is a direct correspondence between the base pairs
sequence in DNA and the amino acid sequence in the corresponding protein.
Nongenetic RNAs involved in protein synthesis
RNA is in intermediary between DNA base pair sequence and amino acid sequence.
Several kinds of RNA have been identified.
RNAs differ from one another in molecular weight, structure and role in protein synthesis.
The macromolecules are measured in Svedberg (S) units, determined by the rate of sedimentation
or the molecule in a density gradient under a standard centrifugal force. The rate of sedimentation
depends on the size and shape of the molecule.
1. Messenger RNA (mRNA)
Messenger RNA is made up of single stranded molecule consisting of nucleotides ranging
from 300 to 12000. In Escheichia coli, mRNA has 900 to 1500 nucleotides. It has a molecular

weight of 4 * 106. It is constituted of the bases adenine, guanine, uracil or cytosine.

In eukaryotes, RNA molecules of variable length with sedimentation coefficient of 20 S to
100 S are involved. This is called heterogeneous nuclear RNA (hnRNA). It is synthesized in the
nucleus and is present in the nucleoplasm outside nucleus. The hnRNA has two fractions, one
with coding sequences that produce mature messenger RNA (mRNA) and another with non
coding sequences that are degraded within the nucleus. The coding sequences of hnRNA
constitute exon and the non coding sequences constitute intron. Polyadenylic acid (poly A)
consisting of about 200 nucleotides is added to hnRNA by the action of polysynthetase enzyme
and after polydenylation, selective degradation takes place by the action of nucleases and the final
product poly A + mRNA reaches cytoplasm to get attached to ribosomes.

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 Functionally mRNA molecules code the sequence of amino acids in protein synthesis.
Eventhough mRNA is single stranded, base pairing occurs between its segments by forming hair
pin loops to produce secondary structures.
Jacob and Monad (1961) first proposed the existence of mRNA.

The life span of mRNA is 2 minutes at 37o C in E. coli, 1 to 4 hours extending to 2 days
in eukaryotes, 6 hours Bacillus cerus and several months and years in the dormant seeds of plants
and animal eggs.
The process by which the information in the nucleotids sequence of DNA is transferred to
a complementary sequence of mRNA is called transcription.

2. Transfer RNA (tRNA)

Transfer RNA molecules are smaller than mRNA molecules and contain 70 to 80

nucleotides. TRNA has a molecular weight of 2.5 * 104 and a sedimentation coefficient of 4 S.
tRNA molecules have a completely folded tertiary structure due to hydrogen bonds between the
constituent bases and due to the presence of a number of unusual bases such as A - U - G - C
situated in the curves of the molecules where no base pairing occurs.
Holley (1968) suggested a clover leaf model of alanine tRNA in yeast with the following
structural peculiarities.
1. An amino acid attachment site with a terminal sequence of CCA at 3' - OH end of the
polynucleotide chain. Adenylic acid (A) is the last residue at 3' end. Its complementary
strand aligned by folding has 5' - P end.
2. T  C arm consisting of a loop with seven unpaired bases including pseudouridine and is
involved in the binding of tRNA molecule to the ribosome.
3. DHU arm consisting of dihydro uridine loop having 8 to 12 unpaired bases and functioning
as the site for recognition of amino acid activating synthetase enzyme.
4. Codon recognition site or anti-codon arm consisting of one nucleotide triplet which is
complementary to the corresponding triplet codon of mRNA.
5. A short extra arm when the chain is long.

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 A three -dimentional L-shaped configuration was proposed by Kim et al. (1972) for phenyl
alanine tRNA molecule of yeast cells.
3. Ribosomal RNA (rRNA)
Ribosomes are ribonucleoprotein molecules present in all types of cells. They occur as 60
S units in mitochondria, 70 S units in bacteria and chloroplast and 80 S units in eukaryotes. Their
constitution is as follows.
Prokaryotic 50 S Sub unit 5 S rRNA, 23 S
ribosome 70 S rRNA + 34 proteins
30 S Sub unit 16 S rRNA + 21
proteins
Eukaryotic ribosome 60 S Sub unit 5 S rRNA, 7 S rRNA
80 S 28 S eRNA + 50
proteins
40 S Sub unit 12 S rRNA + 30
proteins

RRNA is the insoluble RNA that constitutes tha largest part, even up to 80 per cent of the
total cellular RNA. It has four major RNA bases A - G - U - C with a slight degree of methylation.
RRNA molecules are single polynucleotide stranded, unbranched, and flexible and behave
as a random coil or show helical regions with base pairing between A - U and G - C. rRNA has a
definite role in protein synthesis.

Protein Synthesis
Central dogma of molecular biology
The process of protein synthesis involves one of the central dogma of molecular biology,
postulated by Crick (1958) according to which genetic information flows from nucleic acid
to protein.
Protein synthesis involves two steps viz., transcription and translation. Transcription
involves a sequential flow of information from DNA to mRNA. This doesnot involve a change of
code since DNA and mRNA are complementary. Translation involves a change of code from
nucleotide sequences to amino acid sequences.

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 Generally the flow of information is one way, from DNA to RNA and from RNA to
protein.

Transcription Translation
DNA mRNA → Protein

In certain viruses, the existence of an enzymes `RNA dependent DNA polymerase' (also
called inverse transcriptase) was reported and this enzyme could synthesize DNA from a single
stranded RNA template. This finding of Baltimore (1970) and others gave rise to the concept of
`central dogma reverse'. According to this, the sequence of information flow is not necessarily
from DNA to RNA to protein, but can also take place from RNA to DNA.
Transcription Translation
DNA → RNA → protein

inverse transcription

Transcription
The process by which the information in the nucleotide sequence of DNA is transferred to
a complementary sequence of RNA is known as transcription.
Transcription occurs throughout interphase and continues up to early prophase of cell
division. `DNA dependent RNA polymerase' or `transcriptase' is the enzyme involved in
transcription. The locations of transcription are 1) the nucleolus where genes from rRNA are
transcribed and 2) the remaining chromatin where hnRNA (mRNA) is transcribed.
The system for in vitro RNA synthesis contains 1) ribonucleotide triphosphates (ATP,

CTP, GTP and UTP), 2) enzyme RNA polymerase, 3) Mg++ or Mn++ and 4) template DNA.
The enzyme links the ribonucleotides together by catalysing the formation of 3' - 5' phospho diester
bonds that pin the nucleotides. Consequently, RNA is synthesized and pyrophosphate is released.
The enzyme RNA polymerase acts only in the presence of DNA, against which the

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correct sequence of ribonucleotides is arranged and they are linked together by the enzyme. That
is why the enzyme is known as `DNA - dependent RNA polymerase'.
N : r A - P ~ P ~ P (ATP) RNA Polymerase
N : r U - P ~ P ~ P (UTP) + Template DNA →
N : r G - P ~ P ~ P (GTP) Mg++ or Mn++
N : r C - P ~ P ~ P (CTP)
Template DNA + N rA-P
r U - P + 4n P - P
rG-P
rC-P

The site of transcription in cistron is called the promotor site. The template strand is called
sense strand, while its complemetary strand is known as antisense strand. When only one strand of
DNA is transcribed for a given region, it is called asymmetrical transcription. When both the
strands of the DNA are transcribed, it is known as symmetrical transcription.
The details of the transcription process are the following: the enzyme RNA polymerase
attaches itself at the promotor site. The DNA molecule unwinds over a short region. Then the free
bases in the template strand of DNA determine the sequence of ribonucleotides in the newly
formed mRNA.
The RNA polymerase enzyme joins the nucleotides together to produce RNA transcript.
After the transcript becomes detached, the DNA template strand re-forms H-bonds with its
complementary strand rewinds to form the double helix.
RNA polymerase enzyme has five subunits of polypeptide chains viz., , , '  and 
subunits form the core of the enzyme and catalyse the linkage of ribose nucleotides by
phosphodiester bonds. The  - factor recognises the start signal in the promotor region of DNA.

Translation
As soon as the mRNA is formed, it leaves the nucleus and reaches the cytoplasm where
translation takes place.

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 Before the process of protein synthesis, the ribosomes occur in dissociated and inactive
state. The mRNA binds with 30 S ribosomal subunit in the presence of a protein factor called
Initiation Factor (IF). The mRNA carries triplet codons for the synthesis of proteins. Protein
synthesis involves mRNA, ribosomes, amino acids and their specific tRNAs.

Translation process involves the following steps:

1. Attachment of mRNA with 30 S ribosomes and formation of polyribosomes
The mRNA transcribed inside the nucleus moves to the cytoplasm and binds itself with 30
S subunit of the ribosome in the presence of Initiation Factor. Then the tRNA, present in the
cytoplasm binds itself with the first triplet codon 5' - AUG - 3' called the chain initiation codon
of mRNA to form the `Initiation Complex'. Later, the 30 S subunit of ribosome unites with 50 S

subunit to form 70 S ribosome, in the presence of Mg++ ions. The message in the mRNA is not
deciphered by one ribosome but many ribosomes are involved in the process and hence they are
called polyribosomes.
2. Activation of the amino acids
Amino acids present in the cytoplasm ared in a domant stage. Each amino acid is activated
by an activating enzyme called aminoacyl synthetase, beside the energy rich adenosine
triphosphate (ATP). The free amino acids react with ATP to produce aminoacyl adenylate and
pyrophosphate (PP).
Amino acid + ATP + Amino acyl Amino acid - AMP - Enzyme(aminoacyl
synthetase enzyme→ adenylate - enzyme complex) + PP

The aminoacyl adenylate enzyme complex bound together by a mono covalent bond
attaches itself with the specific tRNA molecule. As the enzyme is specific for specific amino acid,
the concerned amino acid gets attached without error.
3. Attachment of activated amino acid to tRNA
The aminoacyl adenylate remains bound with the enzyme till it is hooked to the tRNA
molecule. The dihydrouridine (DHU) loop of tRNA recognises the synthetase enzyme. Then the
amino acid residue of the aminoacyl adenylate is transferred to the amino acid attachment site of

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tRNA, where its carboxyl group forms linkage with 3 - OH group of the ribose of the terminal
adenosine at - CCA end of tRNA.
As a result, adenosine monophosphate (AMP) and the enzyme are released and aminoacyl
tRNA is formed. Then the aminoacyl tRNA moves towards the ribosome.
AA - AMP - Enzyme + tRNA → AA - tRNA +AMP + Enzyme
4. Initiation of the polypeptide chain
In the mRNA, the first triplet codon is AUG at its 5' end. AUG codes for methionine. Hence
protein synthesis commences with coding for methionine. The peptide chain formation starts in
5' end and proceeds towards 3' end and this helps in the correct sequence of protein synthesis.
The mRNA moves across the ribosome. A new codon of mRNA is brought in position. A
new tRNA charged with specific amino acid is brought in position in such a way that the anticodon
of tRNA pairs with the codon of mRNA. The attachment of two amino acids by polypeptide
linkage involves enzymes translocase and peptidyl transferase along with energy rich GTP, and
tRNA is released.
This process of movement of mRNA from 5' to 3' direction and addition of aminoacids to
polypeptide chain continues till mRNA is no longer translated.
5. Termination of the polypeptide chain
Any one of the three terminating codons in mRNA, viz., UAA, UAG or UGA can signal
the temination of chain elongation.
After chain termination, the enzyme peptidyl transferase hydrolyses the ester bond
between the chain and tRNA releasing the polypeptide chain, the last tRNA and mRNA.
Thus a polypeptide chain with a specific series of amino acids is formed which results in
synthesis of a specific protein that involves in a specific phenotypic expression in the organism.

Genetic Code
In the DNA and RNA, there are four types of nucleotides or bases viz., A, G, T, C and A,
G, U, C respectively. If it is assumed that each base codes for one amino acid, then only four amino
acids can be coded. If two bases together are responsible for production of one amino acid, then

they will code for 42 = 16 amino acids. If three bases together code for an amino acid, then

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43 = 64 amino acids could be coded. As the essential amino acids in a biological system are 20 in
number, the possibility of one or two bases coding for each amino acid is remote.
Crick and Brunner (1961) suggested that the genetic code might be a triplet code,
involving three nucleotide bases to code for an amino acid.
Further investigations by Nirenberg and Mathhaei (1961), Nirenberg (1961), Khorana
(1964) and others lead to the construction of a complete genetic code dictionary.

First Second base Third base

base (3' end)
(5'end)
U C A G
U Phe Ser Tyr Cys U
Phe Ser Tyr Cys C
Leu Ser NS NS A
Leu Ser NS Trp G
C Leu Pro His Arg U
Leu Pro His Arg C
Leu Pro Gln Arg A
Leu Pro Gln Arg G
A Ileu Thr Asn Ser U
Ileu Thr Asn Ser C
Ileu Thr Lys Arg A
Met Thr Lys Arg G
G Val Ala Asp Gly U
Val Ala Asp Gly C
Val Ala Glu Gly A
Val Ala Glu Gly G

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 The pattern of genetic code indicates the following:
1. Codons for the aromatic amino acids begin with Uracil
UUU Phenyl alanine (Phe)
UUC
UAC Tyrosine (Tyr)
UAC
UGG Tryptophan (Trp)

2. Codons for amino acids that form amides begin with Guanine and Adenine.
GAU Asparagin (Asp)
GAC
GAA Glutamin (Glu)
GAG

3. For many of the synonymous codons specifying the same amino acids, the first two
bases of the triplet code are constant while the third varies, being less specific.

GCU CUU
GCC CUC
GCA Alanine CUA Leucine (Leu)
GCA (Ala) CUG
GUU CCU
GUC CCC
GUA Valvine CCA Proline (Pro)
GUG (Val) CCG
GGU CGU
GGC CGC
GGA Glycine CGA Arginine (Arg)
GGG (Gly) CGG
ACU UCU

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 ACC UCC
ACA T h r e o n i n e UCA Serine (Ser)
ACG (Thr) UCG

According to Crick (1966), the third base tends to wobble or is unsteady and he
proposed the wobble hypothesis.
The genetic dictionary of mRNA codons reveals the following features of the triplet
code
General feature of genetic code
1. some of the nucleotides have to code for more than one amino acid and hence called
ambiguous code. For example, UUU codes for phenyl alanine and in the presence of
streptomycin, it may code for isoleucine, leucine or serine.
2. The code contains many synonyms and hence called degenerate code. Almost all the
amino acids are represented by more than one codon. For example, arginine, serine and
leucine have six synonymous codons.
For many of the synonymous codons specifying the same amino acid, the first two bases
of the triplet are constant whereas the third varies, as for alanine, valine, glycine, leucine,
proline, arginine, threonine and serine. This flexibility in the third base of codon
minimises the consequences of errors.
3. The code is read continuously without interruption and no codon is reserved for
punctuation. Hence it is called a comma a less code.
4. There is no overlapping of base sequences specifying for different amino acids and no
single base in a triplet can take part in the formation of more than one codon. Hence it is
called a non overlapping code. For example, in a polynucleotide chain, UCAGAA, UCA
codes for serine and GAA for glutamine, but over-lapping as UCA, CAG, AGA etc. does
not occur to code for other amino acids.
5. As the same code applies for all living systems, it is called an universal code. However,
a few codons in some organelle DNAs were found to have different meanings than those
in nuclear DNAs. For example, AUA that normally codes for isoleucine in nuclear tRNA

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 codes for methionine in mitochondrial tRNA and UGA codes for tryptophan in mt tRNA
instead of being a termination codon.
6. Among the triplet codons, AUG is the chain initiation codon as it initiates the synthesis of
polypeptide chain. Codons UAA, UAG and UGA are the terminating codons as they
terminate translation of the polypeptide chain. As these three codons do not specify any
amino acid, they are called nonsense codons.

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