Indian J. Genet., 79(1) Suppl.

340-366 (2019)
DOI: 10.31742/IJGPB.79S.1.27

Genetics on a maize cob: A teaching tool for schools

1,2 3
Firoz Hossain, Shripad Ramachandra Bhat , Trilochan Mohapatra and Ashok Kumar Singh*

1
ICAR-Indian Agricultural Research Institute, New Delhi 110 012; ICAR-National Institute for Plant Biotechnology,
2 3
New Delhi 110 012; XV Genetics Congress Trust, NASC Complex, New Delhi 110 012; Indian Council of
Agricultural Research, Krishi Bhavan, New Delhi 110 001

(Received: February 2019; Revised: April 2019; Accepted: April 2019)

Abstract vaccines, medicines including personalized medicines,

Genetics occupies central position in biology and directly
improved breeds of animals and crops and so on.
connects with all its branches. In fact, ‘life’ and ‘non-life’ Besides, genetic technologies are being increasingly
meet each other in viruses and genes. Technologies used in forensics (Arenas et al. 2017). Recent
emanating from genetics have had immense impact in developments in gene editing (Knott and Doudna 2018)
various areas such as healthcare, agriculture, environment and gene drive technologies (Collins 2018) have raised
etc. Recent developments in stem cell biology and gene
great concern about ethical and moral dimensions of
editing technologies not only promise cures for several
ailments which were hitherto unthinkable but also raise genetics based technologies (Brossard et al. 2019;
concerns of irreparable harm that can result from unbridled Custers et al. 2019). Thus, knowledge of genetics is
use of these technologies. Therefore, basic understanding essential for all citizens as it touches every individual
of genetics is necessary for all sections of society. Hence, in one or more ways.
genetics is now introduced at the school level. However, at
least in the Indian context, genetics teaching in classrooms Schools are the entry points of education system,
relies largely on blackboard or chart-based explanations.
and students’ interest and leanings on subjects are
Unlike physics, chemistry and to some extent basic botany
and zoology where models or live/preserved specimens greatly influenced by the teachers they encounter in
are available to demonstrate fundamental concepts under these early formative years. While text books serve
classroom conditions, there is a dearth of material resources as the primary source of information (substance) to
for teaching genetics. Maize has a long history of basic and school students, it is the style a teacher adopts that
applied genetics where a large number of well-characterized differentiates the best ones from the rest. Those who
mutant stocks are available. To make learning experience
interesting and engaging, we have developed maize genetic
succeed in capturing the attention and imagination of
resources for use under classroom conditions. In particular, students through articulate explanations, including use
maize cob with seeds segregating for various clearly of analogies, anecdotes and effective demonstrations
identifiable phenotypes presents unique opportunity to are invariably regarded as the best teachers. These
demonstrate basic genetic principles including statistical days, genetics is introduced at the primary school level
and population genetic concepts. In this article we describe
alongside other basic subjects such as physics,
these resources and explain how they can be used to
demonstrate different concepts. chemistry, biology and mathematics. In subjects like
physics and chemistry, and to some extent biology,
Key words: Allele, gene, resource, school, student, various tools/models/replicas are available to
model demonstrate the fundamental principles and to
describe the inner workings/mechanisms of different
Introduction
systems under classroom settings. For teaching
Genetics, the study of heredity, is a unifying discipline genetics, however, it is hard to find a model that can
of all branches of biology. Technologies derived from be readily used in a classroom to effectively explain
the understanding of genetics have revolutionized fundamental Mendelian principles. Teachers generally
healthcare and agriculture through novel diagnostics, resort to charts/diagrams for teaching, which not only

*Corresponding author’s e-mail: [email protected]

Published by the Indian Society of Genetics & Plant Breeding, A-Block, F2, First Floor, NASC Complex, IARI P.O., Pusa Campus, New
Delhi 110 012; Online management by www.isgpb.org; indianjournals.com
April, 2019] Maize: A model organism for teaching genetics 341

makes learning less interesting but also leaves most Before introducing genetics, students are already
school-level students confused. Since the past several taught core biology topics such as mitosis, meiosis,
years, we have been actively involved in training sexual reproduction in plants and animals. In fact,
biology school teachers in genetics and biotechnology genetics is best appreciated and easily understood
through workshops organized by the XV Genetics when it is explained using these core concepts. In our
Congress Trust. These interactions have made us experience, failure to connect key aspects of cell
aware of the key problems teachers face in teaching division and seed development with genetic principles
genetics at the entry level, and prompted us to develop is at the heart of much confusion and misunders-
of novel genetic stocks of maize tailor-made for tanding. Mastering Mendelian genetics requires clear
teaching genetic principles to school children. In order understanding of several terms such as ‘cross/self
to bring science lab to classroom, we have developed pollination’, ‘hybridization’, ‘dominance’, ‘recessive’,
‘model genetic resources’ which can be used to ‘gene’, ‘allele’, ‘genotype’, ‘phenotype’, ‘filial
demonstrate more than twenty key aspects/concepts generations’, etc. Further, knowledge of statistical
of genetics. These resources are relevant to school concepts of probability and sampling variation is
as well as college level students. indispensable to comprehend basic laws of inheritance
enunciated by Gregor Johann Mendel (Mendel 1866).
Maize as a model system Therefore, we feel that while introducing these terms,
features of maize especially with respect to its flower
Among various model organisms available, maize and seed development should also be explained.
possesses several unique features for demonstrating
genetic principles. These include i) a wide range of Explanation of key terms
clearly visible mutants for various traits, particularly ‘Self’- and ‘cross-pollination’ (hybridization) are two key
grain traits, are readily available (Neuffer et al. 1997); words in genetics. Pollination is the process of transfer
ii) male and female flowers are borne on separate of pollen grains from anthers to stigma, and is
structures and facilitate crosses; iii) grains on a single facilitated by wind, water, insects, birds etc. Mendel
cob show genetic segregation and represent a described his principles of inheritance using garden
population of individuals; iv) segregating seeds pea which bears hermaphrodite flowers. Textbooks
arranged in rows on a cob can be used to illustrate depicting pea as an example usually show self-
statistical concepts such as random events, sampling pollination as pollination of a flower by its own pollen,
and probability; v) ears can be easily preserved for while cross-pollination as pollination by pollen from
years and readily carried anywhere; vi) the mutants another plant. In contrast to garden pea, maize is a
are well characterized with respect to chromosomal monoecious plant where female inflorescence (called
locations of genes and molecular details of their ear) is borne in the axil, whereas male flowers are
action(s) are well worked out; and vii) people are very borne terminally on an inflorescence called tassel.
familiar with different types of corn (sweet corn, baby Hence, pollen from the tassel needs to disperse and
corn, pop corn, field corn etc.) and explaining their land on silk (fused style and stigma) to effect
differences in terms of genetics, helps to connect fertilization of female flowers. Many students
classroom teaching with day-to-day life experiences. introduced to genetics using pea as the example find
Besides, genetic studies with maize have led to path it difficult to explain cross- and self-pollination when
breaking discoveries such as heterosis (Shull 1948, presented with cases of monoecious or dioecious
1952) and transposable elements (jumping genes) plants. Maize offers good opportunity to clarify that
(McClintock 1951). Efforts have been made in the past self-pollination is not limited to pollination of a flower
to use maize as a model for teaching some concepts by its own pollen but includes all types of matings
of genetics (Chinnici 1999, Ford 2000). Here, we have involving the same or genetically identical individuals,
attempted to expand on those to include additional whereas cross pollination is the mating between
aspects, particularly, concerning statistical aspects. genetically dissimilar (contrasting) individuals. After
We describe below various mutant stocks gathered the students understand this, they will be able to
and assembled by the Maize Genetics Unit, Division explain various types of pollination in dioecious plants
of Genetics, ICAR-Indian Agricultural Research as well.
Institute (IARI), New Delhi and how they could be used
to explain different key concepts of genetics to The terms that should be introduced next are
students. ‘traits/characters’ and ‘character states/forms’ or
342 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

‘phenotypes’. In maize, a large number of mutants and parental derivation. This is particularly important
with contrasting seed features including variations in as maize seeds borne on cobs will be used here to
colour, texture, shape, appearance etc. are available, demonstrate various Mendelian concepts.
which can be very effectively used to explain the
above terms. A complete list of traits and character Maize seed displays typical monocot features.
states investigated by Mendel in pea and available in To understand it fully, one should start from the ovule.
our maize genetic stocks developed for teaching are A specialised cell within the ovule called the
presented in Table 1 and 2, respectively. Further, using ‘megaspore mother cell’ (2n) undergoes meiotic division

Table 1. Details of traits Mendel studied in his pea experiment

S.No. Trait/character Dominant Recessive Target tissue Protein function Chromo-

Allele Character Allele Character (ploidy) some
state state
1. Seed shape R Round r Wrinkled Cotyledon (2n) Starch branching 5
enzyme 1
2. Cotyledon colour I Yellow i Green Cotyledon (2n) Stay-green gene 1
3. Seed coat/flower A Grey/purple a White/white Pericarp/flower bHLH transcription 2
colour (2n) factor
4. Pod colour GP Green gp Yellow Pod (2n) Chloroplast structure 5
in pod wall
5. Pod form V Inflated v Constricted Pod (2n) Sclerenchyma 3
formation in pods
6. Position of flowers FA Axial fa Terminal Stem node (2n) Meristem function 4
7. Stem length LE Tall le Dwarf Stem (2n) GA 3-oxidase1 3
As per Reid and Ross (2011)

examples of mutants in starch biosynthesis, one can to generate four ‘megaspores’ (n) arranged in a linear
also show that phenotype is not restricted to traits fashion (Fig. 1). Of these, the lower three cells (i.e.
that can be seen but also extends to those that could closer to micropyle) degenerate leaving only one
be felt/experienced with other senses like taste. The functional megaspore. This functional megaspore (n)
other term is ‘genotype’. Mendel used the term ‘factor’ undergoes three successive mitotic divisions without
to refer to what we now call gene that determines a cytokinesis to yield an eight-nucleate cell with four
trait. For instance, the factor that makes a plant tall is nuclei each at opposite poles. One nucleus from each
denoted as ‘T’, while its alternate character state, dwarf pole migrates to the centre after which cellularization
is denoted as ‘t’. Mendel used ‘pure breeding’ takes place giving rise to seven-celled embryo sac
(homozygous) lines for hybridization experiments called the megagametophyte. The megagametophyte
which he obtained after repeated cycles of self- has three haploid cells called ‘antipodals’ located at
pollination. The ‘pure breeding’ lines are those whose the chalazal end, while another three haploid cells are
selfed progenies do not show variation for the trait(s) located at the micropylar end. The middle cell of the
under consideration. micropylar triplets is called the ‘egg cell’ while the cells
on its either side are called ‘synergids’. The two nuclei
Before initiating discussion on Mendel’s laws, that migrated to the centre are enclosed in one cell
we feel that a quick recapitulation of angiosperm seed called the ‘central cell’ (polar nuclei) (2n). On the male
development is imperative. As seed comprises of cells side, pollen mother cell (2n) undergoes one meiotic
that are structurally, genetically and compositionally division giving rise to four microspores (n) arranged in
different and unique, while explaining the seed, a tetrad fashion (Fig. 2). These microspores develop
emphasis should be placed on specifying the lineage into pollen grains. During pollen maturation, microspore
and identity of different cell types including their ploidy undergoes one mitotic division to generate two cells
Table 2. Details of nuclear genes that show phenotype in seed and/or cob

S. No. Trait/character Dominant Recessive Target tissue Protein function Chromo-

Allele Character state Allele Character state (ploidy) some
1. Endosperm Ae1 Normal/glossy ae1 Pale Endosperm (3n) Starch branching enzyme IIb (SBE-IIb) 5
April, 2019]

appearance
2. Endosperm Bt2 Round (low sugar) bt2 Highly crumpled Endosperm (3n) Small subunit of ADP-Glucose 4
appearance (high sugar) pyrophosphorylase
3. Endosperm Sh1 Round sh1 Crumpled Endosperm (3n) Sucrose synthase 9
appearance (small dent)
4. Endosperm Sh2 Round (low sugar) sh2 Highly crumpled Endosperm (3n) Large subunit of ADP-Glucose 3
appearance (high sugar) pyrophosphorylase
5. Endosperm Su1 Round (low sugar) su1 Partially crumpled Endosperm (3n) Debranching enzyme (DBE) 4
appearance (high sugar)
6. Endosperm Wx1 Normal/glossy wx1 Pale Endosperm (3n) Granule bound starch synthase (GBSS) 9
appearance
7. Endosperm O2 Normal/glossy o2 Opaque Endosperm (3n) Leucine zip (bZIP) transcription factor of 7
hardness genes coding zein proteins
8. Endosperm Bz1 Purple/red bz1 Bronze Endosperm (3n) & Flavonol 3-o-glucosyltransferase 9
colour depending on Pr1 embryo (2n)
or pr1
9. Endosperm C1 Purple/red depen- c1 Colourless Endosperm (3n) & Transcription factor of genes involved in
colour ding on Pr1 or pr1 embryo (2n) anthocyanin synthesis 9
C1-I Colourless C1 Purple/red depending
on Pr1 or pr1
10. Endosperm Pr1 Purple pr1 Red Endosperm (3n) & Flavonoid 3’-hydroxylase 5
colour embryo (2n)
11. Endosperm R1-nj Purple/red crown r1 Colourless Endosperm (3n) & Transcription factor of genes involved 10
colour colour depending embryo (2n) in anthocyanin synthesis
on Pr1 or pr1
Maize: A model organism for teaching genetics

R1 Purple/red depending r1 Colourless

on Pr1 or pr1
12. Endosperm colour Y1 Yellow/orange y1 White Endosperm (3n) Phytoene synthase 6
13. Haploid induction MATL Fertilization by matl Failure of fertilization Embryo (2n) Pollen specific phospholipase 1
by male gamete male gamete
(diploid embryo, (haploid embryo,
full seed set) partial seed set due
to embryo abortion)
14. Pericarp colour Orp1 White (colourless) orp1 Orange with orp2 Pericarp (2n) -subunit of tryptophan synthase 4
15. Pericarp colour Orp2 White (colourless) orp2 Orange with orp1 Pericarp (2n) -subunit of tryptophan synthase 10
16. Pericarp colour P1-rr Red P1-ww White or colourless Pericarp and cob/ R2R3 Myb-like transcription factor of 1
glume (2n) genes coding maysin
P1-rw Red P1-ww White or colourless Pericarp (2n)
343

P1-wr Red P1-ww White or colourless Cob/glume (2n)

344 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

Fig. 1. Female gametogenesis in plants

Fig. 3. Parts of a maize seed and their chromosome

constitution

embedded) are also maternal tissue with 2n

chromosome constitution.

As per convention in genetics, the first parent

written in a cross is always treated as female, while
the other parent is male. Different ‘filial generations’
(F , F2, BC1 etc.) are the next set of vocabulary of
genetics that needs to be explained. Seeds are
technically progeny of the plant on which they are
borne and thus represent the next generation. Thus,
Fig. 2. Male gametogenesis in plants
seeds borne on a plant after cross pollination are F1,
i.e. the first filial generation. Such F1 seeds give rise
called the ‘generative cell’ and ‘vegetative cell’. to F1 plants. Likewise, seeds borne on F1 plant upon
Generative cell further undergoes one mitotic division self-pollination represent F 2 generation. It is very
to produce two sperm cells (male gametes). One important to clarify that the mother plant and seeds it
sperm (n) fertilizes the egg (n) to produce the embryo produces represent two different, successive
(2n), while the other one (n) fuses with the two polar generations. When the F 1 is crossed to one of its
nuclei (2n) to give rise to endosperm (3n). The ovule parents, it is called backcross and is denoted as BC1F1.
upon maturity develops into seed which is composed Sometimes students are confused with the terms
of endosperm, embryo and pericarp (Fig. 3). Endosperm ‘seed’ and ‘grain’; grain refers to that which is meant
is triploid (3n) where 2n is contributed by the female for consumption whereas seed is that which is meant
parent and n is contributed by the male parent. The for propagation. Thus the two terms are essentially
outermost cell layer of the endosperm is called same from genetics point of view but differ in relation
aleurone which is a living tissue of the endosperm of to their usage. In this article, we have used grains and
mature seed and plays critical role in mobilizing seeds interchangeably.
nutrients to growing seedling. The large portion of
maize seed is made of starchy endosperm. Embryo With these backgrounds, we present below
is diploid (2n) inheriting one set of chromosomes from various genetic principles using maize ears obtained
each of the parents. However, the outer seed coat by selective mating between parents carrying specific
(pericarp) is a maternal tissue and is diploid (2n). Thus, mutations. Thus, a ‘cob’ bearing seeds is called the
it can be said that the mother protects its children ‘ear’. It might be of interest to note here that Mendel
(seeds) wrapped in its own piece of cloth. Besides, tested his pea results with various other plants
the entire plant and cob (pith on which grains are including maize, and in a letter to Carl Wilhelm von
April, 2019] Maize: A model organism for teaching genetics 345

Nageli, the leading Swiss botanist of that time, he

stated that the maize experiments confirmed his
findings with pea (Rhoades 1984). Thus, maize finds
a legitimate place in the list of plants that contributed
to the discovery of fundamental principles of genetics.

Dominance and recessive relationship

Mendel made crosses between pure breeding lines

with seven contrasting character states for different
traits such as plant stature (tall and dwarf), seed shape
(round and wrinkled) etc., to understand the inheritance
of traits (Table 1) (Reid and Ross, 2011). He noticed
that all the F1s invariably exhibited only one of the two
parental character states. However, in the F 2
generation, both the character states were observed
in different individuals. He called the character state
observed in the F1 as ‘dominant’ and the character
state that was suppressed in F1 as ‘recessive’. For Fig. 5. Seed shape obtained by selfing or intermating
example, in a cross between tall and dwarf pea plants, of parents
F1 plant is tall. So, tall is dominant over dwarf and the
factor/allele for the dominant character state ‘tall’ is
round) will carry only round seeds indicating that round
denoted by the upper case letter ‘T’ and the recessive
seed is dominant over shrunken (Figs. 4 and 5). Similar
character state ‘dwarf’ is denoted by the lower case
dominant/recessive relationships can also be observed
letter ‘t’. Thus, the genotype of a true-breeding tall
in our stocks for seed traits such as sugary (Figs. 6
plant is represented as ‘TT’, while the true-breeding
and 7), waxy, opaque, endosperm- and pericarp-colour
dwarf is denoted by ‘tt’. When Mendel made these
and so on (Table 2).
discoveries, chromosomal basis of heredity was
unknown and therefore his designation of genotypes
is indeed praiseworthy.

Using maize ear, dominance/recessive

relationship can be demonstrated for several seed
traits. For example, seed shape (round versus
shrunken) in maize is controlled by the Shrunken2
gene located on chromosome 3 (Hossain et al. 2013;
Mehta et al. 2017a, b). A maize cob obtained by
crossing between pure breeding round- and shrunken-
seeded parents (i.e. round × shrunken or shrunken ×

Fig. 6. Inheritance of seed shape in maize (round

dominant over sugary)

Law of segregation

Mendel’s first law known as ‘law of segregation’ states

that two alleles of a gene co-existing in heterozygous
condition within a cell do not affect each other, and
separate in pure form during gamete formation. For
example, the maize Sugary1 (Su1) gene (located on
chromosome 4) when mutated (i.e. su1 allele) leads
Fig. 4. Inheritance of seed shape in maize (round to accumulation of sugar instead of starch in the grains
dominant over shrunken) (Hossain et al. 2013). As per the law of segregation,
346 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

Fig. 7. Seed shape obtained by selfing or intermating Fig. 9. Segregation of round and sugary in F2 seeds
of parents borne on F1 cob

coming together of alleles Su1/su1 in the F1 does not 3:1 segregation in F2 seeds available in our stocks
lead to their mix-up, and the two alleles separate to include (i) purple: white, (ii) round: shrunken, (iii) normal:
give rise to Su1 and su1 gametes each retaining its waxy, and (iv) normal: high amylose (Fig. 10). Both
unique characteristics. When such gametes unite at
random they will give rise to seeds with three kinds of
genotypes, namely, Su1/Su1, Su1/su1 and su1/su1.
Thus, the hybrid which shows the dominant phenotype
gives rise to selfed progenies that segregate for the
trait in 3 (dominant): 1 (recessive) ratio. For instance,
mutation of the Su1 gene makes the maize seed sweet
and such grains are crumpled at maturity (as against
the round normal seed). This 3 (round): 1 (sugary)
segregation of grains can be clearly seen on a cob
obtained by selfing an F1 plant (Fig. 8). For instance,
in the ear shown in Fig. 9, out of a total 425 grains,
322 are round and 103 are sugary. Other examples of
Fig. 10. Seed trait segregation in F2 seeds borne on F1
cob

sh2 and su1 mutants show shrivelled seeds but the

two phenotypes are clearly distinguishable, indicating
that a trait (seed shape) could be controlled by different
genes. Further, sweet taste of the sugary mutant grain
and its shrivelled shape (at maturity) are two
manifestations of a single gene. This illustrates that a
mutation could alter more than one trait (see later) or
manifest in different ways.

Law of independent assortment

Fig. 8. Checker board analysis of segregation of round Mendel’s second law, known as ‘law of independent
and sugary in F2 seeds assortment’, states that when two or more genes co-
April, 2019] Maize: A model organism for teaching genetics 347

occur in heterozygous condition, segregation of each

gene is independent of the segregation of the other
gene. The implication of this law with complete
dominance is that the F2 derived from an F1 hybrid
whose parents differ for two traits (a dihybrid) will show
9 (dominant/dominant): 3 (dominant/recessive): 3
(recessive/dominant): 1 (recessive/recessive)
segregation when the two traits are jointly considered.
In maize, Waxy1 (Wx1) gene is present on
chromosome 9, while the Yellow1 (Y1) gene is present
on chromosome 6. Seeds with dominant Wx1 allele
produce starch comprising 30% amylose and 70%
amylopectin, while grains with only recessive wx1
Fig. 12. Independent assortment of yellow/white and
alleles accumulate 95-100% amylopectin, thereby glossy/waxy F2 seeds borne on F1 cob
impart a dull look to the endosperm compared with
glossy endosperm of normal maize grains (Devi et al.
backcross. The test cross progenies show 1:1
2017, Hossain et al. 2019). Likewise, the dominant Y1
segregation for the trait under consideration whereas
allele produces carotenoids giving yellow/orange colour
in the other backcross (i.e. F1 × dominant parent), all
to the endosperm, while its recessive allele (y1) fails
progenies will display dominant phenotype. Testcross
to produce any carotenoids and thereby gives white
thus allows identification of zygosity (homozygous or
endosperm (Muthusamy et al. 2014; Zunjare et al. 2017,
heterozygous) status of an individual showing dominant
2018). When an inbred with yellow-glossy grains is
phenotype. The simplified segregation ratio (3:1 in F2
crossed with white-waxy inbred, the F2 seeds show 9
versus 1:1 in testcross) means smaller sample size
(yellow-glossy): 3 (yellow-waxy): 3 (white-glossy): 1
could be used to test the segregation ratios (see later).
(white-waxy) ratio (Fig. 11). Note that each of the trait,
In maize, several examples of 1:1 segregation can be
demonstrated in testcrosses (Fig. 13). F 1 between

Fig. 11. Checker board analysis of independent

assortment of yellow/white and glossy/waxy
Fig. 13. Segregation of round and shrunken seeds in
traits in F2 seeds
testcross and backcross

endosperm colour (yellow versus white) and grain normal inbred (Sh2Sh2) and shrunken (sh2sh2) when
appearance (glossy versus waxy), considered singly crossed with the recessive parent (sh2sh2), the seeds
shows 3:1 segregation (Fig. 12). These results on an on F1 cob showed 176 (round): 168 (shrunken) (i.e.
ear beautifully demonstrate that Y1 and Wx1 genes BC1F1 generation) thereby exhibiting 1:1 segregation
segregate independent of each other. (Fig. 14). While the F1 crossed with dominant parent
(Sh2Sh2) had all round grains.
Testcross
Maternal effect
Testcross is the cross between F1 and its parent that
carries the recessive allele. Thus, it is in essence a In maternal effect, the phenotype of the progeny is
348 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

(as female) and P1-rw/ P1-rw (as male) generates F1

seeds with white pericarp, thereby exhibiting reciprocal
difference. The F1 plants (P1-rw/P1-ww) of both the
crosses upon selfing yield F2 seeds with red pericarp
only (Fig. 16). Thus F2 seeds on a cob do not show

Fig. 14. Seed shape inheritance in testcross and

backcross

determined by the genotype of the maternal parent.

Coiling (right handed or left handed) of shell in snail is
often cited in text books to explain the maternal effect.
In maize, maternal effect can be demonstrated by
considering the pericarp colour of the seed. Dominant
Pericarp colour1 (P1-rw) allele located on chromosome Fig. 16. No segregation for pericarp colour in F2 seeds
1 imparts red colour to the pericarp due to the borne on F1 cob
accumulation of pigment (phlobaphenes), while no
pigment accumulates when its recessive mutant allele segregation for the pericarp colour unlike other traits
p1-ww is present in homozygous condition thereby such as sugary, waxy, shrunken etc. This is best
giving transparent pericarp (Goettel and Messing 2010). explained by considering the tissues involved in
Transparent pericarp reveals underlying endosperm expression of these traits. Pericarp of the F2 seeds is
(yellow, white or purple) thereby displaying different diploid maternal tissue (2n) and has the same genotype
coloured seeds depending on the genetic constitution as the F1 plant (P1-rw/p1-ww), whereas other traits
of the endosperm. A cross between P1-rw/P1-rw (as such as sugary, waxy, shrunken etc. are endosperm
female) and P1-ww/P1-ww (as male) leads to formation traits which is a triploid tissue (3n) derived from the
of F1 seeds (borne on female plant) with red pericarp fusion of central cell (2n) and sperm cell (n), and
(Fig. 15). While the reciprocal cross, P1-ww/P1-ww belongs to the F2 generation. Segregation for pericarp
colour can be seen among cobs (bearing F3 seeds)
borne by F2 plants, but in no case segregation for
pericarp colour is observed among seeds on a single
cob (Fig. 17). It is worth noting that Mendel encountered
differential behaviour where seed shape and cotyledon
colour showed segregation within a pod, while seed
coat colour did not. He presented these results in his
lectures but avoided including the seed colour in his
1866 publication (Zhang et al. 2017). The fact that
Mendel could correctly interpret these results at a time
when meiosis and double fertilization in plants were
unknown attests to his extraordinary intelligence and
insight.

Maternal effect is also seen for traits such as

Fig. 15. Inheritance of pericarp colour in F1 seeds of glucosinolate and fatty content in mustard seeds. In
direct and reciprocal crosses this case, unlike the pericarp colour of maize, these
April, 2019] Maize: A model organism for teaching genetics 349

Fig. 17. Inheritance of pericarp colour in maize in direct and reciprocal crosses

metabolites are present in cotyledons which are the classified plants into contrasting categories such as
products of fertilization. Therefore, maternal tissue of tall and dwarf, and counted the number of individuals
the seed cannot be used to explain maternal effect. of the two categories in segregating generations. He
In Brassica, glucosinolates are synthesized in leaves did not resort to measuring height of individual plants
and silique wall, and are transported to seeds as practised by his contemporaries working to unravel
(cotyledons). Hence, all seeds on a plant irrespective the mystery of inheritance. This mathematical
of their genotype show glucosinolate content and treatment allowed Mendel to work out the proportion
profile corresponding to their mother. In the case of of the two categories and thereby arrive at the laws of
fatty acids, however, synthesis takes place in the inheritance. Size of the population is an important
cotyledons but the photosynthates necessary for fatty factor for drawing conclusion as small populations
acid synthesis are supplied from the mother plant which could show highly distorted ratios.
allocates equal amount to all its progenies. It is to be
noted that total fatty acid content shows maternal Toss of a coin or roll of dice is commonly used
effect but not the fatty acid profile (i.e. types of fatty to demonstrate randomness of occurrence of events
acids) which shows segregation among seeds on a and the concept of probability in genetics teaching
plant. The practical implication of maternal effect is (Lesnik 2018). Maize ear serves as an excellent model
that genotyping and phenotyping need to be done, to illustrate random occurrence of segregation and the
respectively, in successive generations. Thus, effect of population size on segregation ratio. Maize
maternal effect is of great practical significance and ear generally possesses 12-14 rows and each row has
calls for careful analysis and explanation to avoid 30-40 grains. When students are presented with an
confusion (see cytoplasmic inheritance later). ear segregating for say, normal and sugary seeds,
they will be able see the random distribution of two
Random events, sample size and segregation ratio types of seeds along a row and find that no two rows
show identical pattern of distribution of normal and
Mendel’s success in cracking the basis of inheritance sugary seeds. Also, when the students are asked to
of traits is often attributed to his mathematical calculate segregation ratios row wise, and on whole
approach (Zhang et al. 2017). In other words, Mendel cob basis, they will be able to realise the effect of
350 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

sample size on ratio. An example of segregation of (Morgan 1911). A pair of genes/traits are said to be
normal versus sugary grains is presented in Table 2 completely linked if the F 2 progenies show only
and Fig. 18. The random distribution of normal and parental types (trait combinations present in the
parents). However, if the linkage is incomplete, F 2
progenies will show a large fraction of parental types
and a small fraction of recombinant phenotypes (trait
combinations not present in the parents). It is now
established that the proportion of recombinants
depends on the distance between the two genes. The
discovery of linkage has led to the demonstration that
genes are linearly arranged on chromosomes and
thereby led to chromosomal basis of heredity.

Although Thomas Hunt Morgan is rightly credited

with the discovery of linkage based on his experiments
with Drosophila, Carl Correns, one of the three persons
who independently rediscovered Mendel’s principles,
Fig. 18. Effect of sample size on F2 segregation ratio had perhaps encountered linkage as early as 1902
of round and sugary seeds (Rhoades 1984) in his studies with maize (sugary and
starchy grain traits). However, he could not correctly
sugary seeds is clearly evident. Also, it can be seen interpret his results. In maize, genes controlling seed
that in some of the rows segregation ratio deviates shape and stem base colour are linked where dominant
widely from 3:1, but when whole ear is considered the alleles Sh2 and Anthocyanin1 (A1) (both located on
ratio approaches 3:1 and improves further when more chromosome 3) conferring ‘round seed’ and ‘purple
ears (say three) are considered for calculating the ratio. stem base’, respectively occur together, while
Whenever small samples are taken, sampling variation recessive alleles (sh2 and a1) conferring ‘shrunken
often leads to deviation from the expected ratio. The seed’ and ‘green stem base’ occur together. This type
genetic segregation ratio in Mendel’s experiments fitted of association where dominant alleles of two or more
well with the expected ratio of 3:1 and 9:3:3:1 in genes are on the same chromosome, while their
monohybrid and dihybrid crosses, respectively (Mendel recessive counterparts on the other homologous
1866). As a matter of fact, very close concordance chromosome, is known as ‘coupling phase linkage’.
between observed ratios and expected frequencies in In ‘repulsion phase linkage’, dominant allele of one
Mendel’s report has raised doubts about truthfulness gene is present with recessive allele of another gene
of his data (Weeden, 2016). The near perfect fit of on one homologous chromosome whereas their
observed data with that of expectation in Mendel’s opposite alleles are found on the other homologous
studies could be attributed to large data set used by chromosome. Sh2 is physically located at a short
him. In general, whenever samples are taken, there is distance (140 kb, <1cM) from A1. In most maize
possibility of conscious or unconscious bias of the accessions, the recessive alleles sh2 and a1 occur
experimenter in sampling and calculating ratios. together while the dominant alleles Sh2 and A1 occur
However, use of maize ear segregating for seed traits together (coupling phase linkage). In a cross between,
can overcome the problem of experimenter’s bias Sh2/Sh2/A1/A1 and sh2/sh2/a1/a1 plants, F2 seeds
as the number of seeds on a cob and their segregation segregate for round and shrunken in 3:1 ratio (Fig.
in different rows is governed by random events of 19). When the seeds are allowed to germinate, all
nature. seedlings from round seeds possess purple
pigmentation at the stem base, while all shrunken
Linkage
seeds give rise to seedlings which lack purple pigment
Mendel arrived at ‘law of independent assortment’ at the base (Chhabra et al. 2019). The purple versus
based on study of only seven traits in pea. Subsequent green stem base trait also shows 3:1 segregation in
findings brought to light that this law is not applicable F2. Since the distance between the two genes is quite
in all cases and some traits tend to inherit together short, complete linkage is observed when limited
(linked). Thus, linkage is defined as the tendency of number (100-200) of F2 plants are examined (Fig. 20).
two or more genes/traits to be inherited together However, in a larger population (1000-5000), a few
April, 2019] Maize: A model organism for teaching genetics 351

on F1 cob showing the segregation of both the genes.

The effect of incomplete linkage can be

demonstrated even in a small population on a single
ear of corn when segregation of Bronze1 (Bz1) and
Shrunken1 (Sh1) genes are considered together. Both
the genes are present on chromosome 9 in close
vicinity (~2.5 cM distance). The dominant Bz1 leads
to formation of anthocyanin pigments (purple colour)
in the endosperm when A1, C1 and other genes of the
pathway are also present in dominant condition.
Presence of recessive bz1 alleles yields bronze colour
in place of typical purple colour conditioned by Bz1.
Fig. 19. Checker board analysis of complete linkage The dominant Sh1 gives round seeds, while the
between seed shape and stem pigmentation
recessive sh1 creates small dent (depression) in the
in F2
crown area of the grain. When, Bz1Bz1/Sh1Sh1 (purple
and round) is crossed with bz1bz1/sh1sh1 (bronze and
shrunken), the majority of F2 grains are of parental
type, while a few grains show recombinant phenotypes
(purple and shrunken; bronze and round). We are in
the process of developing these stocks for
demonstration.

Pleiotropy

When mutation of a single gene affects more than

one trait, it is called pleiotropy. Maize provides several
examples of pleiotropy. Among various genes
responsible for nutritional enhancement in maize (Das
et al. 2019a, b, Muthusamy et al. 2014), Opaque2
Fig. 20. Complete linkage between seed shape (round (O2) gene located on chromosome 7 determines lysine
and shrunken) in F 2 seeds and stem and tryptophan content of grains (Mertz et al. 1964).
pigmentation (purple and green) in F2 plants Traditional maize grains with the dominant O2 allele
accumulate low amount of lysine (1.5-2.0% of protein)
recombinant types (round grains with green stem base, and tryptophan (0.3-0.4% of protein) whereas lysine
and shrunken grains with purple stem base) can be (3.0-4.0% of protein) and tryptophan (0.7-0.9% of
also observed. The synthesis of purple pigment requires protein) content is almost double in grains carrying
not only A1 but also another gene Coloured aleurone1 only recessive o2 alleles (Fig. 21). Besides altered
(C1) located on chromosome 9 in dominant condition protein content, o2 grains are soft and do not let light
(Sharma et al. 2011). We have developed inbreds with pass through when placed on a light box; hence the
Sh2Sh2/A1/A1/C1C1 (round and purple grains) and name ‘opaque’ (Hosain et al. 2008a, Hossain et al.
sh2sh2/a1/a1/C1C1 (shrunken and yellow grains) 2018, Sarika et al. 2018a). This opaqueness is due to
genetic constitution. The F2 seeds obtained from the loose packaging of protein bodies in the endosperm.
cross between these inbreds show only parental In contrast, protein bodies are tightly packed in normal
phenotypes but not recombinant types (round and maize grains which allow the light to pass through
yellow, shrunken and purple). Thus, due to tight linkage giving them a translucent look. Thus, both
between A1 and Sh2 genes located on chromosome accumulation of lysine/tryptophan and grain softness
3, Sh2Sh2/a1a1/C1C1 (round and yellow) and sh2sh2/ are conditioned by the recessive o2 gene (Fig. 21).
A1A1/C1C1 (shrunken and purple) are not produced. Pleiotropy in this case arises due to cascading effect
Thus complete linkage (when small population size is of seed protein composition on packaging of nutrients
considered) can very well be demonstrated in a single in a grain. Another good example of pleiotropy in maize
ear itself. We have developed the parental stocks, is the sugary mutant. Su1 mutation makes maize
and currently in the process of generating F2 grains grains sweet and such grains are shrivelled like raisins
352 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

pigments) but in two different tissues. Interestingly,

pea seed coat colour gene (A) studied by Mendel is
also responsible for flower colour, and is a case of
pleiotropy (Reid and Ross, 2011). Although Mendel
observed strict association between these two traits,
he omitted these two characters from his 1866
publication (Zhang et al. 2017).

Multiple alleles

Mendel’s pea examples had only two alternative

character states for each trait and hence two alleles
for each gene. Subsequently, it was found that a gene
could have more than two alternative forms (i.e.
multiple alleles) each displaying a distinct phenotype.
Fig. 21. Pleiotropic effects of opaque2 gene. A: higher Since a diploid individual can have only two copies of
lysine and tryptophan levels in QPM (o2o2) over a gene, all alleles of a gene with multiple alleles cannot
normal maize (O2O2). B: Ear of normal maize, occur in a single individual. Instead, such multiple
C: ear of QPM, D: grains of normal maize on
alleles occur in different combinations among
light box, E: grains of QPM on light box
individuals of a population. In the previous section,
we have discussed P1 gene governing pericarp and
at maturity. Grains with su1 allele (i.e. endosperm su1/ glume colour of the cob. More than 100 natural alleles
su1/su1) are impaired in starch biosynthesis and of P1 have been reported (Sekhon and Chopra 2009).
therefore accumulate more sugars giving sweet taste Of these, four alleles viz., P1-rr, P1-rw, P1-wr and p1-
to grains. Such grains on maturity tend to crumple ww are commonly observed and phenotypically
and hence appear shrivelled. Note that the sh2 mutation distinguishable (Goettel and Messing 2010). The first
also gives shrivelled seeds with enhanced sweetness letter after P1 indicates colour of seed pericarp (r =
of the grain, but its phenotype is distinct from su1 and red, w = white/no colour), and the second letter
is easily distinguishable. These examples also indicates colour of the glumes on cob. The allele, p1-
illustrate that apparently similar phenotypes could have ww is recessive allele to all the other three alleles.
different genetic underpinnings. P1-rr gene responsible Thus, plants with P1-rr/P1-rr show red colour in both
for red pericarp colour also imparts red colour to the pericarp and glume, P1-rw/P1-rw plants have red
glumes (Fig. 22). Thus, P1-rr gene is also pleiotropic; coloured pericarp but white glumes, P1-wr/P1-wr plants
however, pleiotropy in this case results from a gene have white coloured pericarp but red glume, while the
performing the same function (production of red recessive p1-ww/p1-ww lacks colouration in both
pericarp and glume (Fig. 22). Thus, P1 gene serves to
illustrate two concepts, pleiotropy and multiple alleles.
It is interesting to note that mutations in the P1 gene
affect expression of the trait in different tissues
(regulatory sequence mutations) and thus yield multiple
alleles and cause pleiotropy. It also demonstrates
dominance relations are hierarchical among different
alleles.

Penetrance

An individual carrying a gene/allele does not always

show the corresponding phenotype because phenotype
results from gene expression, and gene expression
can be influenced by the environment and/or modifier
genes present in the background. The term
‘penetrance’ indicates the likelihood of an individual
Fig. 22. Variation in pericarp and glume/cob colour due
to multiple alleles of P1 gene who carries the allele also displays the corresponding
April, 2019] Maize: A model organism for teaching genetics 353

phenotype. If all individuals carrying a given allele show ‘variable expressivity’. For instance, all maize seeds
the expected phenotype, it is complete penetrance. possessing the recessive o2 allele show opaque
Various seed phenotypes discussed earlier such as phenotype (complete penetrance) (Vasal et al. 1980,
waxy, sugary, shrunken, opaque are examples of Gupta et al. 2013). However, the degree of opaqueness
complete penetrance. Incomplete penetrance could is variable among grains on an ear segregating for the
result from environmental effects or the presence of trait (Fig. 24) which is clearly visible when grains are
other interacting genes elsewhere in the genome. In
such case, only a fraction of individuals carrying the
allele show the corresponding phenotype. In maize,
pollen expressed Matrilineal (MATL) gene located on
chromosome 1 mediates fertilization of male gamete
with the egg cell (Kelliher et al. 2017). When maize
plant carrying normal MATL allele is crossed with pollen
from matl matl plants, fertilization takes place but the
chromosomes from the male parent fail to propagate
during zygotic divisions leading to production of
maternal haploid embryos. However, an inbred
homozygous for matl/matl upon crossing produces
haploids in only 8-10% of the seeds (~10%
penetrance). Besides haploids, mat1 mutation causes Fig. 24. Variable expressivity for kernel opaqueness
embryo abortion in a few developing seeds in selfed conditioned by recessive o2 allele. A: ear of
ears of matl/matl plants (Fig. 23) as against well-filled normal maize, B: ear of QPM, C: variable
degree of opaqueness in seeds of an ear of
grains found when the dominant allele (MATL) is
QPM inbred, D: variation of softness in QPM
present. seeds (F ) (orange and yellow regions depict
hardness and softness, respectively), E: F 2
seeds viewed under light box , black indicates
soft region, translucent represents hardness

observed under a light box (Hossain et al. 2008a).

Regions where light does not pass through appears
as black (thus opaque) and indicates soft region, while
the region that permits light to pass through appears
as orange and represents the hard region (Hossain et
al. 2007; Sarika et al. 2018b). With experience, one
can also visualize the varying degree of opaqueness
just by looking at grains. This variation is more profound
when inbreds with different genetic backgrounds are
compared. Such varying shades of a phenotype are
ascribed to (i) presence/absence of modifier genes in
Fig. 23. Incomplete penetrance for embryo abortion by
matl allele different genetic backgrounds (Pandey et al. 2015),
and/or (ii) influence of environmental factors on gene
expression. Expressivity is particularly important in
Expressivity the context of genetic ailments where low expressivity
and low penetrance would greatly lessen the symptoms
Expressivity is a measure of the degree of expression
and suffering.
of a trait in a set of individuals who carry the allele and
also display the phenotype. When the degree of Dosage effect
expression is similar in individuals carrying the target
gene, it is called as ‘uniform expressivity’. P1-rr and Dosage effect is observed when trait expression varies
P1-rw possess red colour in the pericarp with uniform depending on the number of copies of specific alleles
intensity and expression, thus having ‘uniform in a genotype. For example, in maize Red1-navajo
expressivity’ (Fig. 22). When a gene shows variable (R1-nj) located on chromosome 10 imparts purple
degree of expression among individuals, it is called pigmentation in the crown of the endosperm (Fig. 25)
354 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

Fig. 25. Dosage effect of R1-nj allele (in F seeds borne Fig. 26. Activity of transposable elements (TEs)
on F1 cob) conditioning purple pigmentation in affecting pericarp (A) and endosperm (B-D)
crown regions of endosperm pigmentation

(Chaikam et al. 2015). The recessive r1 allele fails to colour of the endosperm is interrupted, it does not
produce purple colour in the crown region of produce any colour. The excision of TE restores the
endosperm. Selfing of a heterozygous plant (R1-nj/r1) function of C1 leading to the formation of purple
yields four types of seeds, namely, deep purple, light pigment patches (Fig. 26B-D). However, depending
purple, fade purple and no pigment with corresponding upon the time of excision of TE from the C1 gene,
endosperm genotypes of R1-nj/R1-nj/R1-nj, R1-nj/R1- different sizes of deep purple pigmented patches are
nj/r1, R1-nj/r1/r1 and r1/r1/r1 (Fig. 25). Therefore, R1- observed. Early excision leads to large coloured spot,
nj allele displays dosage effect with increasing number while late excision leads to smaller spots. Thus, a
of dominant alleles giving more intense pigmentation. complex concept of jumping gene in action could be
Similarly, Y1 gene producing yellow/orange endosperm easily demonstrated with our maize stocks.
colour also shows dosage effect (Fig. 12). Endosperm
with Y1/Y1/Y1 is deep yellow/orange, Y1/Y1/y1 is light Xenia
yellow/orange, Y1/y1/y1 is fade yellow/orange, while If a seed trait (e.g. colour, size, shape etc.) is
y1/y1/y1 is white with no synthesis of carotenoids. influenced by the pollen carrying the dominant allele,
Transposable elements (Jumping gene) it is called xenia. For instance, when a sh2/sh2 maize
plant (recessive homozygous) receives Sh2 (dominant
The position of a gene on a chromosome is fixed and allele) pollen, the resulting seed will have Sh2/sh2/
it is known as ‘locus’. Working with maize, McClintock sh2 endosperm constitution and will form round instead
(1951) reported unusual behaviour of genes controlling of shrunken seeds (sh2/sh2/sh2 endosperm) expected
seed colour which appeared to switch on and off within from self pollination (Fig. 27). Thus, a few round grains
a seed giving various seed colour patterns. Detailed
examination led to the discovery of Transposable
Elements (TEs) as the cause of this behaviour. These
TEs are capable of moving from one place to other on
the same or different chromosomes. When a TE moves
and inserts into a gene, it may disrupt the function of
that gene, while its excision restores gene function.
Maize genome possesses several highly active TEs
such as Ac-Ds MuDR-DMu, Spm-dSpm (En-I) and
Dt1-dt1, which impart unusual colour pattern to grains.
For example, P1-rw gene imparts red colouration to
the pericarp but when it is interrupted by a TE, it does
not produce red coloured pericarp (Fig. 26A). However,
when TE moves out of the P1-vv gene, it restores its
function and starts producing the red colour in stripes. Fig. 27. Xenia effects of foreign pollens bearing
Similarly, when C1 gene responsible for deep purple dominant allele on different seed traits
April, 2019] Maize: A model organism for teaching genetics 355

among other shrunken grains indicate contamination by F1 plant and not in F1 seeds produced on the parent.
displaying xenia effect. Similar effect is also found Heterosis can be measured in three ways. When the
when Y1 pollen fertilizes y1/y1 plant (Y1/y1/y1: yellow, performance of F1 is compared with its mid-parent
y1/y1/y1: white), O2 fertilizes o2o2 plant (O2/o2/o2: value (average of the two parents), it is called ‘average
Glossy, o2/o2/o2: dull/opaque) (Hossain et al. 2008b) heterosis’, while comparison of F1 with the better of
and C1 fertilizes c1c1 plant (C1/c1/c1: purple, c1/c1/ the two parents, it is called ‘heterobeltiosis’. When
c1: colourless endosperm). It is of historical interest the performance of F1 is judged against commercial
to note that de Vries and Correns independently variety/hybrid, it is called as ‘economic heterosis’.
noticed xenia effect working with maize seed traits Maize lines show rapid decline in performance for
starchy/sugary and coloured/colourless endosperm almost all traits over successive generations of selfing.
(Rhoades 1984). One may note that xenia and maternal This phenomenon is called ‘inbreeding depression’.
effects are contrasting features. Traits showing Hybrid breeding involves developing true breeding
maternal effect are not affected by xenia and vice inbred lines (homozygous) and then identifying the best
versa. Such xenia effect is of practical significance combination of inbreds that gives highly heterotic
especially in cross pollinated crops where stray pollen hybrid. ‘Pusa Vivek QPM-9 Improved’ is a single cross
from open pollination could affect nutritional quality hybrid developed by IARI, New Delhi. The parents are
traits conditioned by recessive alleles. Recessive o2 PMI-PV-1 and PMI-PV-2. The average weight of a
gene based quality protein maize (QPM) hybrid is an single ear of the hybrid is 220 g, while the ears of
example. As discussed earlier, in mustard, parents are 60 g and 80 g, respectively (Fig. 28) and
glucosinolate content and profile are maternally the comparable commercial hybrid (Vivek Maize
controlled and are not affected by open pollination. Hybrid-27) gives ears with mean weight of 200 g. Thus,
However, fatty acid profile (but not fatty acid content) the three types of hererosis can be calculated as below.
of seeds shows xenia effect. Performance of F1-mean
Heterosis and inbreeding depression performance of parents
Average heterosis = x 100%
Heterosis is defined as the superior performance of Mean performance of parents
F1 over its parents/check variety. The concept of
heterosis was first proposed by East and Shull based 220  (60  80) / 2
= x 100%
on their studies with inbreeding of maize (Rhoades (60  80) / 2
1984). Heterosis is now widely exploited to obtain higher
yields in crops through breeding of hybrid cultivars, = 214%
and forms the foundation of hybrid seed industry. Performance of F1-
Heterosis can be vividly demonstrated in a classroom performance of better parent
by presenting maize ears produced on parental inbred Heterobeltosis = x 100%
lines and their hybrids (Fig. 28). It is important to Performance of better parent
emphasize that heterosis is realized on ear produced
220  80
= x 100%
80
= 175%

Performance of F1-
performance of commercial
hybrid
Economic heterosis = x 100%

Performance of commercial hybrid

220  200
= x 100%
200
Fig. 28. Ears of parents and their hybrid depicting
heterosis for ear length, seed number and ear = 10%
weight
356 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

Unlike Mendel’s traits where specific phenotypes deep purple, light purple, fade purple and no pigment
are associated with one or a few genes, yield heterosis have diploid genotypic (in embryo) constitution as R1-
is governed by many genes. Although each of these nj/R1-nj, R1-nj/r1, r1/R1-nj and r1/r1, respectively. Using
genes follow Mendel’s laws of inheritance, discrete ears of different generations produced through open
classes are not observed because each gene makes pollination among different mixtures of individuals
a small contribution towards phenotypic expression segregating for R1-nj and/or Y1 genes one can
of one or more traits which ultimately lead to heterosis. demonstrate Hardy-Weinberg principle to students. For
Further, environmental factors also influence trait example, consider a sample of 430 grains drawn from
expression thereby confound the detection of individual the harvest of a large random mating population have
contribution of genes towards a trait. four genotypic classes as 135 R1-nj/R1-nj, 120 R1-nj/
r1, 155 r1/R1-nj and 20 r1/r1. Since, R1-nj/r1 and r1/
Hardy-Weinberg Law R1-nj represent heterozygote, the frequency of
Hardy and Weinberg independently proposed a law heterozygote class is (120+155)/430 = 0.64. Similarly,
pertaining to frequency of occurrence of alleles and the frequency of homozygotes R1-nj/R1-nj and r1/r1
genotypes over generations in a population that is 135/430 = 0.31 and 20/430 = 0.05, respectively.
reproduces through random mating among individuals. Thus the gene/allele frequency of R1-nj = 0.31 + (0.64/
The law states that allele and genotype frequency will 2) = 0.63, and r1 = 0.05 + (0.64/2) = 0.37. Now, we
remain constant over generations in a large random can calculate the genotype frequency of the next
2
mating population in the absence of disruptive forces generation as R1-nj/R1-nj = (0.63) = 0.40, R1-nj/r1 =
2
such as mutation, migration, selection and random 2×0.63×0.37 = 0.47, r1/r1 = (0.37) = 0.13. It shows
drift (Hardy 1908, Weinberg 1908). Allele frequency is that either the the initial population was not in
also referred to as ‘gene frequency’ and ‘gametic’ equilibrium or the sample does not truely represent
frequency. Random mating means that an individual the population. In any case, after one generation of
in a population has an equal opportunity of mating random mating of individuals of this sample, the gene/
with any other individual of the same population. This allele frequency will be (R1-nj = 0.63, and r1 = 0.37)
law is central to all population genetics and is usually and genotype frequency will be (R1-nj/R1-nj = 0.40,
explained using algebraic equations. To calculate allele R1-nj/r1 = 0.47 and r1/r1 = 0.13) and this will remain
and genotype frequencies, one has to determine the so in subsequent generations if they are maintained
genotype of individuals. As explained earlier, by random mating, and other disruptive forces are not
dominance obscures identification of homozygous and operating.
heterozygous individuals expressing the dominant Hardy and Weinberg equilibrium can also be
phenotype. Using a conventional testcross to effectively applied in case of traits where dominant
determine zygosity status is quite demanding and homozygotes and heterozygotes cannot be
hence the operation of this law in nature is rarely distinguished based on phenotype. In a sweet corn
demonstrated. These days molecular markers could composite (population), it was found that out of
be used for this purpose but in this article we confine randomly selected 500 grains, round and shrunken
to features that can be used in classroom without using grains were 50 and 450, respectively. We now know,
sophisticated techniques. Again, maize ears offer a that sh2sh2sh2 endosperm gives shrunken phenotype,
chance. while Sh2Sh2sh2, Sh2sh2sh2 and Sh2Sh2Sh2
In previous pages, we explained how gene condition gives round phenotype. These four
dosage effect influences several endosperm colour endosperm classes correspond to sh2sh2, Sh2sh2,
traits. For instance, each of the endosperm phenotypes sh2Sh2 and Sh2Sh2 embryo (diploid) genotypes. Thus
such as deep purple, light purple, fade purple and no frequency of recessive sh2sh2 is = 450/500 = 0.90.
pigment conditioned by R1-nj gene could be The frequency of Sh2– is = 1-0.90 = 0.10. The
unambiguously assigned to respective genotypes (Fig. frequency of sh2 allele = 0.95 (square root of 0.90).
25). Note that based on endosperm genotype one could Hence, the frequency of dominant Sh2 = 1-0.95 = 0.05.
also clearly specify the male and female gamete. For This calculation has practical significance, as sweet
instance, heterozygous endosperm genotype R1-nj/ corn population should always have recessive sh2,
R1-nj/r1 results from the union of R1-nj female with r1 any contamination by dominant Sh2 allele in the
pollen while R1-nj/r1/r1 occurs from the fertilization r1 population will reduce the grain quality as Sh2 carrying
female with R1-nj pollen. Thus, the four types of grains grains will not be sweet. Since maize is a cross
April, 2019] Maize: A model organism for teaching genetics 357

pollinated crop, foreign pollen from neighbouring field reactions is converted to amylose and amylopectin,
can always contaminate and deteriorate the quality. the components of storage starch. Maize grains
The present example suggests that though frequency normally possess 30% amylose and 70% amylopectin.
of dominant Sh2 is quite low, selection should be Grains desiccate during maturity and take the round
practised to remove the round grains during selection shape due to packaging of starch and protein.
of ears for raising the next generation of sweet corn However, the mutant sh2 allele impairs conversion of
population. UDP-glucose to ADP-glucose, leading to less
synthesis of starch. Thus sh2 grains have less dry
Connecting phenotype and genotype matter while the sink size (grain volume) remains same
The classical Mendelian genetics did not provide a as the normal Sh2 grain. As a result, at maturity such
clear link between genotype and phenotype. However, grains collapse or shrivel as they lose water. These
molecular biology studies of gene action have provided grains with mutant sh2 allele possess high sugar and
us with a complete picture of information flow from are harvested at 20 days after pollination to be sold
gene to phenotype for several traits. The central as sweet corn. Another such example is the su1 gene
dogma of molecular biology where information flows located on chromosome 4. The dominant Su1 codes
from DNA to RNA to protein in a linear fashion (Crick for starch debranching enzyme and is responsible for
1970) holds good in most cases although some synthesis of amylopectin (Fig. 29). The mutant su1
exceptions have been well recognised. Most often, it allele affects this conversion step, leading to less
is the protein that a gene codes for determines the starch synthesis. However, the effect is less severe
phenotype. In maize, various examples clearly depict than that seen with sh2 allele. Hence, su1 mutant grains
such flow of information. The Sh2 gene located on appear as partially shrivelled at maturity. When both
chromosome 3 codes for the large subunit of ADP- the genes are in recessive condition (i.e. sh2sh2/
glucose pyrophosphorylase enzyme that catalyzes su1su1) the grains show extreme crumpling (Mehta et
the conversion of UDP-glucose to ADP-glucose (Fig. al. 2017c). The Wx1 gene located on chromosome 9
29). ADP-glucose by a further series of biochemical encodes granule bound starch synthase enzyme

Fig. 29. Role of Sh2 and Su1 genes in starch biosynthesis (as per Whitt et al. 2002) and their effects on seed
phenotype
358 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

Fig. 30. Role of Wx1 and Ae1 genes in starch biosynthesis (as per Whitt et al. 2002) and their effects on seed
phenotype

responsible for the synthesis of amylose (Fig. 30). gene that masks the effect of the other gene is called
The recessive wx1 allele almost completely blocks ‘epistatic’ while the gene that is masked is referred to
this step, leading to accumulation of 95-100% as ‘hypostatic’. Considering that a gene regulates just
amylopectin. The grains with high amylopectin look one step in a series of reactions of a long biochemical
pale and can be easily identified from the normal glossy pathway leading to the manifestation of a trait, it is
grains. Further, the recessive amylose extender1 (ae1) easy to visualise occurrence of epistasis among genes
enhances the amylose level from 30% in normal grains regulating a metabolic pathway. The first hint of
to a higher level (50-70%, average: 60%) (Stinard et epistasis is that a known gene fails to show expected
al. 1993). The grains of ae1ae1 can be easily identified segregation in a new genetic background. At least six
by their pale look in comparison to glossy appearance different kinds of epistatic interactions involving two
in the normal maize. These examples demonstrate genes are commonly recognised which give distinct
how information flows from gene to phenotype. The F2 segregation ratios (Table 3).
molecular function and phenotype of various other
genes have been mentioned in Table 2. Epistasis is best illustrated with grain colour
variations in maize. Apart from pericarp colour
Epistasis
(maternal tissue), maize grain colour is determined by
Epistasis is intergenic interaction involving two or more anthocyanins and carotenoids accumulating in the
genes where one gene masks the effect of the other endosperm (Ford 2000). Biosynthesis pathways for
gene(s). This is akin to dominance/recessive anthocyanins (Sharma et al. 2011) and carotenoids
relationship between alleles of a gene (intragenic) giving (Naqvi et al. 2011) are well characterized. Maize
different character states that we have discussed endosperm could be bronze, red, purple, depending
above; in this case, however, the phenotypic change on the type of anthocyanins that accumulate there, or
occurs due to interaction between different genes. The colourless when no anthocyanins accumulate. In the
April, 2019] Maize: A model organism for teaching genetics 359

Table 3. Commonly observed F2 ratios in digenic epistatic interactions

S.No. Epistasis F2 phenotype ratio Genotypes in each phenotype class

1. Dominant epistasis 12:3:1 [9: A-/B- + 3: A-/bb] : [3: aa/B-] : [1 aa/bb]
2. Recessive epistasis 9:3:4 [9: A-/B-] : [3: aa/B-] : [3: A-/bb + 1 aa/bb]
3. Duplicate genes with cumulative effect 9:6:1 [9: A-/B-] : [3: aa/B- + 3: A-/bb] : [1 aa/bb]
4. Complementary epistasis 9:7 [9: A-/B-] : [3: aa/B- + 3: A-/bb + 1 aa/bb]
5. Duplicate dominant genes 15:1 [9: A-/B- + 3: aa/B- + 3: A-/bb] : [1 aa/bb]
6. Inhibitory epistasis 13:3 [9: A-/B- + 3: A-/bb + 1 aa/bb] : [3: aa/B-]
*A and B are unlinked genes but exhibit interactions

absence of anthocyanins, the carotenoid pigments in presented in Fig. 31 to facilitate illustration of various
the endosperm becomes visible and hence the grains epistastic interactions. Pr1 codes for flavonoid 3-
could be yellow or white depending on whether or not hydroxylase and drives the pathway downward to
it accumulates carotenoid pigments (Zunjare et al. produce purple pigment, cyanidin (Sharma et al. 2011).
2017, Goswami et al. 2018). Thus, when pericarp is When Pr1 function is lost (i.e. pr1 allele), endosperm
colourless, grains could be bronze, red, purple, yellow appears red due to the accumulation of pelargonidin
or white. pigment. The other important genes of the pathway,
namely A1, A2, Bz1 and Bz2 also code for key
Anthocyanin biosynthesis in grains involves enzymes of the pathway (structural genes). Genes
several steps and at least seven genes are well C1 and Red1 (R1) code for transcription factors and
characterized in maize. Simplified pathways are are essential for expression of A1 gene and other genes

Fig. 31. Biosynthesis pathways of A: anthocyanin (as per Sharma et al. 2011), B: carotenoid (as per Naqvi et al.
2011) in maize
360 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

in the pathway. Loss of function in any of the genes in Duplicate dominant genes
the anthocyanin pathway arrests the pathway giving
colourless endosperm. Y1 gene which functions in the When a gene is duplicated, loss of function of one
endosperm codes for phytoene synthase and is gene will be compensated by the other. In maize,
responsible for yellow colour of the endosperm and its Orange pericarp1 (Orp1) and Orp2 are duplicate genes
loss-of-function gives white endosperm. present on chromosome 4 and 10, respectively.
Tryptophan synthase is composed of - and -
To understand various epistatic interactions, we subunits, while both Orp1 and Orp2 code for -subunit
have to consider joint segregation of a pair of these (Wright et al. 1992, Fig. 32). The double recessive
genes (i.e. dihybrid ratio) when the rest of the genes
of the pathway are homozygous for the dominant allele
(and carry the recessive allele p1-ww for pericarp
colour). Note that these genes are not linked and
segregate independently.

Dominant epistasis

Here the dominant allele at one locus masks the effect

of the other gene. The F1 hybrid with R1r1/Y1y1 (purple
grains) represents a case of dominant epistasis. Here,
the R1 allele imparts purple colour to the endosperm
and thereby masks the yellow/white colour of the
starchy endosperm. As a result, in the F2, 12 (purple):
3 (yellow): 1 (white) segregation will be found as 9 R1-
/Y1- and 3 R1-/y1y1 will be purple, 3 r1r1/Y1- will be
yellow and 1 r1r1/y1y1 will be white.
Fig. 32. Tryptophan biosynthesis pathway in maize (as
Recessive epistasis per Wright et al. 1992)

As the name suggests, here the homozygous

recessive condition at one locus masks the effect of (orp1orp1/orp2orp2) is unable to convert anthranilate
the gene at the second locus. This is exemplified by and indole into tryptophan. Double mutant grains
the F1 hybrid with Pr1pr1/R1r1 (purple grains). In the develop intense orange pigmentation in the pericarp
F2, this dihybrid segregates as 9 (purple): 3 (red) and and germinated seedlings die at 4-5 leaf stage. The
4 (yellow) because 9 Pr1-/R1- will be purple, 3 pr1pr1/ dominant alleles of both the genes produce transparent
R1- will be red, 3 Pr1-/r1r1 + 1 pr1pr/1r1r1 will be yellow pericarp. The F1 hybrid (Orp1orp1/Orp2orp2) produces
(Y1 allele is present in homozygous condition). Here, 15 yellow (9 Orp1-/Orp2- + 3 Orp1-/orp2orp2 + 3
the recessive condition at the R1 locus masks the orp1orp1/Orp2-): 1 orange (1 orp1orp1/orp2orp2) in the
effect of the dominant Pr1 allele. This is also called genetic background of Y1 allele.
supplementary gene action.
Duplicate genes with cumulative effect
Complementary epistasis
In this case, two duplicate genes possess cumulative
This occurs when two dominant alleles of a pair of effect. Specific alleles of two genes interact to give a
genes are necessary to express a trait. Consider an new phenotype while each of the single gene exhibits
example where the F1 hybrid is C1c1/R1r1. Note that identical phenotypes. The other genotypic condition
C1 is dominant over c1 and R1 is dominant over r1. of both the genes gives third type of phenotype. It is
Thus, the F1 will have purple grains but the F 2 will also called duplicate complementary epistasis.
show 9 [C1-/R1-] purple: 7 [3 C1-/r1r1 + 3 c1c1/R1- + Example of duplicate complementary epistasis is
1 c1c1/r1r1] yellow segregation for grains colour in encountered in the F1 (Sh2sh2/Bt2bt2). Sh2 codes for
the background of Y1 allele. This is also called duplicate large subunit of ADP-glucose pyrophosphorylase, while
recessive epistasis because functions of two genes Brittle2 (Bt2) located on chromosome 4 encodes
are necessary for expression of a trait while any one smaller subunit of the same enzyme (Whitt et al. 2002,
fails to do so. Fig. 29). While dominant alleles of both the genes
April, 2019] Maize: A model organism for teaching genetics 361

exhibit round phenotype, their recessive allele gives In plants, the majority of imprinted genes are
crumpled phenotype. In F2 grains, 9 round (Sh2-/ expressed during early stages of seed development,
Bt2-): 6 crumpled (3 sh2sh2/Bt2- + 3 Sh2-/bt2bt2): 1 particularly in the endosperm. Depending on the
extremely crumpled (sh2sh2/bt2bt2) are observed. The parental origin of the functional allele, imprinting is
crumpling of grains caused by sh2sh2 and bt2bt2 are classified as ‘maternal’ or ‘paternal’. In maize,
phenotypically similar, but the double recessive imprinting was first reported by Kermicle (1970) for R1
(sh2sh2/bt2bt2) is highly crumpled and can be allele responsible for red coloured endosperm. When
distinguished from crumpling caused by either sh2 or R1 allele is inherited from the female parent, the
bt2. endosperm (R1/R1/R1 or R1/R1/r1) will be uniformly
red. However, if the R1 allele is received from the
Inhibitory epistasis male parent, the endosperm (r1/r1/R1) will be mottled
We have previously discussed multiple alleles. The with irregular patches of red. It is pertinent to state
C1 locus has a third allele designated as C1-I (C1- that only specific alleles of R show imprinting. It is
Inhibitor) with the dominance relations as C1-I dominant now known that such epigenetic changes result from
over C1, while both being dominant over c1. C1-I parent-specific acetylation/methylation of chromatin
inhibits the C1 allele. Now consider an F1 hybrid with bound histones or DNA leading to suppression of
C1-IC1/R1r1. This F1 will show yellow grain colour as transcription (Gehring and Satyaki 2017).
the action of C1 allele is inhibited in the presence of Paramutation and epialleles
C1-I allele. Upon selfing, this hybrid will show 13 yellow
[9 C1-I-/R1- + 3 C1-I-/r1r1+1 C1C1/r1r1]: 3 purple As per Mendel’s first law, alleles in a hybrid do not
[C1C1/R1-] segregation in the genetic background of affect each other and are passed on to the next
Y1 allele. This is also called dominant and recessive generation retaining their characteristic features.
gene interaction. Exception to this rule was first reported by Brink (1956)
in maize, where some alleles of the R1 gene when
Epistasis with three genes brought together with a functional R1 allele were found
to induce heritable suppression of the functional allele.
When dominant alleles of two genes give rise to a
This phenomenon is called ‘paramutation’; the allele
new phenotype, it is called complementary epistasis.
responsible for alteration of expression is called
Note that epistasis may involve more than two genes.
‘paramutagenic’ (designated as R1) while the allele
Complementary epistasis in maize grains can be
that is altered is called ‘paramutable allele’. In fact,
demonstrated with the F1 trihybrid Pr1pr1/C1c1/Y1y1.
once altered, the paramutable allele itself becomes
This F1 will show purple grains due to the presence of
paramutagenic and is designated as R1* to distinguish
Pr1 and C1 alleles. In the F2, four types of grain colours
it from R1. An important feature of this phenomenon
namely, purple, red, yellow and white will be found in
is that the two alleles do not show any change in the
the ratio of 36: 12: 12: 4 (or 9: 3: 3: 1). Note that the
nucleotide sequence of the gene. It is to be noted that
trihybrid will produce 27 Pr1-/C1-/Y1- (purple), 9 Pr1-/
most R1 alleles are neither paramutagenic nor
C1-/y1y1 (purple), 9 Pr1-/c1c1/Y1- (yellow), 9 pr1pr1/
paramutable (i.e. neutral alleles). Thus, when such
C1-/Y1- (red), 3 Pr1-/c1c1/y1y1 (white), 3 pr1pr1/C1-/
paramutable and paramutagenic alleles are brought
y1y1 (red), 3 pr1pr1/c1c1/Y1- (yellow) and 1 pr1pr1/
together in a hybrid, grains become colourless or lightly
c1c1/y1y1 (white) classes.
coloured as anthocyanin synthesis will be attenuated.
Genetic imprinting Further, F2 grains do not show any segregation. The
P1-rr gene that we discussed earlier (Fig. 22) also has
For most genes, parent-of-origin of an allele does not paramutagenic alleles P1-rr. Paramutation has been
affect its expression i.e., an allele behaves identically described for total five genes in maize, namely, R1,
irrespective of whether it is inherited from the male or P1, B1, Pl1 and lpa1-241 (Arteaga-Vazquez and
the female parent. For example, we have seen that Chandler 2010) of which, all except lpa1-241 code for
both (Su1/Su1 × su1/su1) and (su1/su1 × Su1/Su1) transcription factors that regulate genes governing
crosses give normal F1 (Su1/su1) grains (dominant biosynthesis of purple and red pigments in different
phenotype) indicating that Su1 allele coming from either parts of maize plant. Molecular studies of paramutation
of the parents is equally effective. However, for some have revealed that the altered allelic expression pattern
genes, the expression of an allele is determined by its (suppression) is brought about by changes in cytosine
parent-of-origin. Such genes are said to be imprinted. methylation of DNA by RNAi-mediated gene silencing
362 Firoz Hossain et al. [Vol. 79, No. (1) Suppl.

mechanisms (Arteaga-Vazquez and Chandler 2010).

Such heritable changes in gene expression

without associated changes at the nucleotide level of
the concerned gene, is called ‘epigenetics’ and the
altered allelic state is called epiallele. Epigenetic
changes in gene expression induced by environmental
factors have now been discovered for many important
developmental and agronomic traits. In maize, tissue
culture was found to produce epialleles of the P1-wr
allele (Fig. 22), which showed partial to complete loss
of pink colour in glumes (Rhee et al. 2010). P1-rw
epiallele shows high methylation in the second intron
which leads to suppression of P1-rw. We are planning Fig. 33. No segregation of cytoplasmic genes in direct
to procure these paramutagenic and epialleles for and reciprocal crosses. Shaded and white
cytoplasm indicate different genetic
inclusion in our kit to demonstrate these concepts to
constitution
students.

Cytoplasmic inheritance (S: USDA) (Allen et al. 2007). The chimeric sequence
Turf13 results in CMS-T, while chimeric sequence of
Soon after the rediscovery of Mendel’s laws in the
three genes, viz., atp6, atp9 and cox-III are found in
beginning of the twentieth century, case of uniparental
CMS-C. Similarly, ORF355 and ORF77 chimeric
(maternal) inheritance was reported. It is now well
sequences are associated with CMS-S system (Table
established that genetic material is present not only
4). When a male sterile parent is crossed with fertile
in the nucleus but also in cytoplasmic organelles,
parent, the F1 carries CMS cytoplasm contributed by
plastids and mitochondria. Mendel’s laws operate for
the female parent, and therefore fails to produce pollens
the nuclear genes, while mostly uniparental inheritance
is observed for cytoplasmic genes. Unlike nuclear
Table 4. Details of maize cytoplasm types causing
genes which occur in two copies per cell (diploid), maternally inherited male sterility
organelle genes occur in multiple copies (100-1000/
cell). In most angiosperms, pollen delivers only sperm S.No. Cytoplasm Mitochondrial gene Target
nuclei and very little cytoplasm during double causing male sterility tissue
fertilization, cytoplasmic organelles and genes are (ploidy)
inherited from the mother only. This is therefore called 1. CMS-T Turf13 Pollen (n)
‘maternal inheritance’. Like maternal effect, maternal 2. CMS-C atp6, atp9 and cox-III Pollen (n)
inheritance show reciprocal difference in F 1, but no
3. CMS-S ORF355 and ORF77 Pollen (n)
segregation is observed for traits controlled by
organellar genes (Fig. 33) in any generation. The key
difference between ‘maternal effect’ and ‘maternal
inheritance’ is in the location of genes controlling the
trait; maternal effect genes are located in nucleus,
while cytoplasmic inheritance is governed by genes
located in mitochondria or plastids.

Cytoplasmic male sterility (CMS) is a well known

maternally inherited trait in maize and other crops which
is governed by mitochondrial genes. In fact, CMS trait
is widely used for large scale production of hybrid
seeds in several crops as it overcomes the need for
manual emasculation of plants to be used as the
female parent. Based on mutation in different causal
Fig. 34. A: Tassel of normal maize showing exerted
genes in cytoplasm, maize CMS is categorized into
anthers shedding pollen, B: tassel of Texas
CMS-T (T: Texas), CMS-C (C: Charrua) and CMS-S male sterile line lacking anthers
April, 2019] Maize: A model organism for teaching genetics 363

leading to complete male sterility. In male sterile plants, Acknowledgements

anthers do not emerge from the glumes (Fig. 34). The
manifestation of male sterility is the formation of barren We thank Indian Council of Agricultural Research
cobs (cob without seeds) where no chance of foreign (ICAR) for funding the In-House and CRP projects.
pollen contamination from neighbouring field exists. Thanks are also due to Department of Biotechnology
CMS demonstration can be done by growing maize (DBT) for funding projects under which some of the
plants in pots. Further fixed tassels can be used to genetic resources have been developed. We thank
show this phenotype. ICAR-Indian Agricultural Research Institute, New Delhi
for providing necessary support in developing the
genetic materials under the maize programme. Support
Conclusion of AICRP-Maize, India; CIMMYT, Mexico; Maize
Training lectures organized for school teachers by ‘XV Genetic Cooperation Stock Centre (MGCSC), USA and
Genetics Congress Trust’ made us realize the need United States Department of Agriculture (USDA) for
for developing resources for easy understanding of providing some of the source germplasm are thankfully
genetics at school level. In recent years, a variety of acknowledged. We thank Drs. Vignesh Muthusamy
approaches have been advocated (Burhansstipanov and Rajkumar Zunjare for their support in maintaining
et al. 2001; Pukkila. 2004; Smith and Wood 2016; these novel maize germplasm. We are grateful to XV
Deutch 2018) and material resources (Arabidopsis, Genetics Congress Trust for providing opportunity to
Price et al. 2018), (pea, Kudish et al. 2015), molecular participate in their outreach programme to train school
tools and other resources (Haga 2006) have been teachers in genetics and biotechnology.
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