2.

2 Morphology of Cereals

The embryo or germ of cereal grains is enclosed in a fragile structure being rich in protein and
enclose the new-fangled plant. A special packing of cells known as scutellum differentiate the
germ from endosperm and play a key role in providing energy to the germinating seeds. The
endosperm is composed of thin-walled cells which are packed within starch granules. Each
cereal grain has specific layers of aleurone cells surrounding the endosperm; wheat, maize,
sorghum, oat and rye constitute one cell layer while rice and barley possess three cell layers. The
pericarp originating from the ovary of flower constitutes the outer layers of grain and encases
seed coat while the external thick walled structure forms the bran. The spikelets constitute the ear
or spike of grain. In Poaceae, pericarp and seed coat are present in combined form. Thus, this
type of fruit is a typical feature of cereals and botanically termed as caryopsis. Cereal flowers
develop into a dry and monocot fruits, known as “grain” or “kernel”. There is a great diversity in
the weight and size of the grains e.g. corn grains is about 350 mg while millet grain is ~9 mg
(Evers and Miller 2002).

2.2.1 Wheat

Wheat Triticum aestivum L. belongs to the family Poaceae, subfamily Pooideae and tribe
Triticeae. It has various synonyms and common names as wheat, bread wheat and common
wheat. Bread wheat is an allohexaploid (6x), that has 21 pairs of chromosomes (2n = 42) during
meiotic cell division. These chromosomes are further categorized into 3 closely connected
(homoeologous) set of chromosomes designated as the A, B and D genomes. All of these
homoeologous groups usually include 7 pairs of chromosomes (AABBDD) (Dvorak et al. 1998;
Haudry et al. 2007).
Wheat kernel termed as “caryopsis” constitutes bran (12-14%), germ (2-3%) & the endosperm
(83%). The bran is made up of pericarp, testa and the aleurone layers. The endosperm and germ
are separated by the scetulum. The kernel is normally 5-9mm long, has an average weight of 35-
50mg and possesses a crease on one side of grain from where it is initially joined to the flower.
The cells present in the bran and aleurone layers are composed of live protoplast and these are
rich in proteins & enzymes. These nutrient dense cell layers have a crucial role during the
germination process (Belderok et al. 2000).
2.2.2 Rice

Rice is the second largest produced cereal crop after maize across the globe. Botanically it is
known as Oryza sativa L. and belongs to the tribe oryzeae. Oryzeae includes 12 genera and about
22 species. Among them, 20 species are wild and 2 Oryza sativa & Oryza glaberrima are grown
for food purposes. Rice has diploid (2n=24) or tetraploid genome (4n=48) and designated as AA
and BBCC (Brar and Khush 2003). The caryopsis of rice grain is 5-12mm long & 2-3mm thick,
doesn’t have a crease and comprises of glumes, endosperm and embryo (Vaughan and
Marishima 2003). Awns are present at the tips of the grain in some cultivars. These may become
a hurdle in dehusking so must be removed prior to dehulling (Vaughan et al. 2003). Li et al.
(2000) illustrated caryopsis (brown rice) and hull as main parts of the kernel. Hull weight is
approximately 20% of the whole kernel weight and composed of lemma & palea. Caryopsis
constitutes the germ (2-3%), endosperm (89-94%) and numerous thin layers of distinguished
cells such as; the pericarp (2%), the seed coat & nuclellus (5%). The seed coat (bran) constitutes
six layers of tissues in which the aleurone layer is the innermost. The main part of rice kernel is
endosperm which is composed of starch, protein, fat, sugar, crude fiber & total ash. The starch
granules are embedded in the protein matrix. Rice varieties vary in their kernel length, width &
thickness (Moldenhauer and Gibbons 2003).

2.2.3 Maize
Botanically termed as Zea mays L. and also known as corn, is an economical source of energy in
the form of biofuel and an easily accessible resource of animal feed and its products like starch,
which is utilized in numerous valuable industrial purposes (Macrae et al.1993; Fast and Caldwell
2000). Maize belongs to the tribe Andropogoneae having 86 genera with Zea including 5 species
with haploid number of chromosomes 2x=2n=20 and the genome is designated as AA (Ellneskog
et al. 2007). There is a great diversity among the corn cultivars regarding the composition of
endosperm and kernels. The four commercially important classes of maize include, firstly, dent
maize (identified by hard endosperm on sides & base of the kernel and when dried dent produces
on the top due to contraction of soft endosperm, grown for silage) secondly, flint maize (hard
endosperm with a small soft centre, use as food), thirdly, sweet corn (dent type maize with
elevated level of moisture & sugars) and fourthly, popcorn (flint type maize i.e. hard endosperm
which expands upon heating) (Paliwal 2000c)
The maize grain is composed of four parts as: the germ, the pericarp, the endosperm and the tip
cap. Corn is a monocotyledonous grain and constitutes a single seeded fruit along with some
loosely attached layers of the seed coat and nuclellus. The grain is connected to the ear through
the pedicel which remains attached to the bottom of the kernel. Maize kernel constitutes 5-6% of
the pericarp, 10-14% germ while the remaining part is the endosperm (Liu et al. 2007).

2.2.4 Barley

Hordeum vulgare L. belonging to tribe Triticeae is a cool season, fast growing, primitive cereal
crop commonly cultivated for food, feed and brewing purpose (Hughes and Baxter 2001).
Hordeum is unusual in the aspect that it has both annual & perennial species i.e. H. Vulgare &
H. marinum are annual while H. bulbosom is perennial. The genus hordeum includes 32 species
all having 7 number of chromosomes (x=7) with diploid (2n=2x=14), tetraploid (2n=4x=28) and
hexaploid (2n=6x=42) species (Komatsuda et al. 1999; Von Bothmer et al. 1999). Barley has a
great diversity in its types; two & six rowed barley, hulled & naked barley. The floret of a barley
plant has a spike at the top of the stem and it composes of 2-6 rows of fertile spikelets which
ultimately develop into mature caryopsis. A layer of cells termed as husk is present as outer layer
of grain and compose of lemma & palea. The hull of grain is tightly bound to its pericarp. The
kernel is composed of pericarp, seed coat, germ, aleurone layers & endosperm. Two to three
layers of cells are present in the aleurone layers with some varieties having coloured aleurone
(blue) layers (Kumar and Goh 2000).

2.2.5 Sorghum

Sorghum bicolour L. Moench is a temperate climate crop, belonging to the tribe Andropogoneae.
Bicolour is most pronounced genera having 25 species which are further categorized into five
subspecies as bicolour, guinea, caudatum, kafir and durra. Sorghum is unable to tolerate low
temperature conditions but has moderate resistance towards serious pests and diseases (Zohary
and Hopf 2000). It possesses diploid & tetraploid number of chromosomes (2n=2x=20;
2n=4x=48). Sorghum grain is spherical in shape and weighs about 20-30mg. Development of
seed occurs in three stages: milk dough, early dough & late dough. It is usually classified on the
basis of its end use as: (1) grain sorghum; (2) forage sorghum; (3) grass sorghum; (4) Sudan
sorghums & broomcorn (Serna and Rooney 1994). Sorghum grains are also classified on the
basis of their distinguishing characteristics such as: pericarp colour & thickness, colour of testa,
colour & type of endosperm. The caryopsis of sorghum grain is composed of three parts:
pericarp (7.9%), germ (9.8%) and endosperm (82.3%). Some varieties have hard endosperm
while others have soft. The endosperm provides required nutrients to the germinating seedlings.
Pericarp of the kernel constitutes three different layers such as epicarp, mesocarp & endocarp.
Along with seed coat, grains have another small cap called as “glume”. Sorghum varieties vary
on the basis of colour of seed coat & glume, although removed through dehulling but sometimes
may present in flour and affect its colour. These colour components include certain
phytochemicals i.e. condensed tannins that may exert some beneficial role such as an antioxidant
or lower digestibility of protein and iron (Rooney and McDonough 1987; Macrae et al. 1993).

2.2.6 Millet

Millet comprises of a number of divergent species that are annual cereal grasses with small
rounded grains (Macrae et al. 1993). Among all the diversified species, the most significant is the
pearl millet botanically known as pennisetum glaucum and tribe paniceae. Some other minor
species of millet include finger millet (Eleusine coracana), porso millet (Panicum
scorbiculatum) & foxtail millet (Digitaria exilis). Millet is not as much essential as other cereal
grains in terms of global food production whereas, human consumption accounts about less than
1 % of all the cereal grains (Andrews and Kumar 1992). It possesses tetraploid number of
chromosomes 2n=4x=36. The caryopsis weighs about 9 mg and comprises of pericarp, germ and
endosperm. The germ constitutes 17% of the kernel while the remaining part is the endosperm
(Rooney and McDonough 1987).

2.2.7 Rye

Rye botanically termed as Secale cereale L. is a member Poaceae in the tribe Triticeae. Cereale
possess diploid number of chromosome as 2n=2x=14. It is usually employed in bread making,
brewing and animal feed. The kernel is mostly hull less and has crease just like wheat grain but
is longer and thinner than wheat. The caryopsis encloses the bran, germ and endosperm (~86% of
the grain) with bran comprising the pericarp and the seed coat and constitutes about 10% of the
grain (Shewry and Bechtel 2001).
2.2.8 Oat
Oat (Avena sativa L.) belongs to tribe Aveneae in the family Poaceae. It includes 3 pronounced
species with chromosome number as 2n=2x=14 in the diploid species, 2n=4x=28 in the
tetraploids, and 2n=6x=42 in the hexaploids. The caryopsis is enclosed by the glumes (hull; layer
of lemma & palea) that remain attached to it. Hull constitutes 25% of the total kernel weight. The
caryopsis is termed as “groat”, which constitutes about 65 to 85 % of the kernel and enclosed by
pericarp, seed coat and aleurone cells of the bran. One third part of the groat is covered by germ,
other is the endosperm. The oat groat is rich in fat and protein as compared to the other cereals
(Kent and Evers 1994).

2.2.9 Pseudo cereals

Along with the cereal grains, some promising cereal like grains termed as pseudo cereals are also
gaining momentum owing to their starchy endosperm, belongs to the family Chenopodiaceae
with three grains i.e. quinoa (Chenopodium quinoa), amaranth (Amaranthus hypochondriacus)
and buckwheat (Fagopyrum esculentum) as mentioned in the “International AACC list of
recognized grains”. Pseudo cereals belong to the dicot family and have resemblance to cereal
crops in terms of their chemical composition and functional properties possessing splendid
nutritional components of proteins, minerals and vitamins. Thus, can be a good source of
alternative crops to supplement food products for enhancing their nutritive value (Brennan et al.
2012). Several phylogenetic classifications depict that Amaranthus and Chenopodium genus
belongs to Caryphyllales while Fagopyrum belongs to Polygonales (Drzewiecki et al. 2003). The
scanning electron micrograph of these grains reveals that they are smaller in size as compared to
the cereal grains and the embryo is enveloped in a layer of starchy tissue termed as perisperm
(Alvarez-Jubete et al. 2010). The heath promoting components known as “bioactive moieties”
are an essential constituents of pseudo cereals such as phagopyritols, phytosterols, polyphenols,
saponins and squalene (Berghofer and Schoenlechner 2002).

The diversified chemical composition of these grains makes them suitable for functional
properties such as the high amylopectin content makes them stable towards freezing, thawing
and retrogradation. Likewise, amaranth protein concentrates exhibit better solubility, foaming
and emulsifying properties (Fidantsi and Doxastakis 2001). These are now considered as a well
known source of gluten free cereals. Quinoa and amaranth are used for the development of a
wide variety of gluten free cereal products (Taylor and Parker 2002).
2.3 Chemical Composition
Cereal crops are usually consumed as a staple diet across the globe. In addition to their food
uses these are consumed as animal feed and non-renewable energy source in the form of biofuel
& bioethanol. Cereals have great variability in their chemical composition like all other food
groups. Compositional analysis of cereal grain has revealed the presence of essential nutrients
for a balance diet such as protein, dietary fiber and lipid (Fincher and Stone 1986; Truswell
2002). These components also affect the end use quality of cereal grains in food and feed.
Cereals are the energy packets constituting all the macro and micronutrients in a well-defined
ratio. On dry matter basis, carbohydrates (56-74%) are the major constituents followed by
proteins (8-11%), lipids (2-4%) and minerals & vitamins (1-3%), respectively (Table 2.1). The
components are located in different parts of cereal grains such as endosperm is the source of
starch and protein while dietary fiber, minerals and vitamins are mostly present in germ and
bran, respectively varying among different cereals in their respective varieties (Wrigley and
Batey 2010).

2.3.1 Carbohydrates

In cereal grains, 66-76% of their weight is carbohydrates that is a diverse group composed of
major and minor nutrient; starch (55-70%) and other non-starch polysaccharides (NSP).
Arabinoxylans (1.5–8.0%) and β-glucans (0.5-7.0%) while oligosaccharides in the form of
glucofructans (~1-3%) are the key examples (Goesaert et al. 2005).

2.3.1.1 Starch

It is the main storage carbohydrate of cereal grains (instant source of energy) and has
diversified functional and nutritional properties especially in bread and other baked products
(Zeeman et al. 2010). Starch is basically a polysaccharide composed of glucose subunits that are
held together by α, 1-4 & 1-6 glycosidic linkages. It is a heteropolymer with two distinct
polymer chains termed as amylose and amylopectin. Amylose is a linear polymer having α, 1-4
linkage with a degree of polymerization (DP) of 1000-1500 glucose residues. Amylopectin is a
branched polymer with α 1-4 &1-6 bonding. Starch is present in the form of granules in the
endosperm of different cereals with variable size such as rice starch granule has a diameter of 5
μm and wheat has 25-40 μm, shape (large, small, round or spherical) and content of amylose and
amylopectin fractions of starch. Generally, cereal starches consist of 25-28% amylose and 72-
75% amylopectin, while their mutant genotypes have varied amylose/amylopectin ratio. “Waxy”
varieties may contain up to 100% amylopectin level whereas “high amylose” cultivars have very
high amylose (70%) content. Variability in the ratio of amylose/amylopectin can alter their
functional attributes. High amylose starches have more resistant starch content than normal
cultivars and have physiological function as they resist the action of digestive enzymes and act as
dietary fiber (Hizukuri 1996; Van Hung et al. 2006). The fraction of dietary starch that is not
hydrolysed by the endogenous enzymes of human intestine is called as the resistant starch (RS).
Four different types of resistant starches; Type 1(RS1) physically inaccessible starch, Type 2
(RS2) native starch granules, Type 3 (RS 3) retrograded starch and Type 4 (RS 4) chemically
modified starches have been classified by Englyst et al. (1992). RS has been recognized as a
splendid substrate for the fermentation of microorganisms present in the colon and is valuable
due to its high levels of short chain fatty acid production. Animal trials have shown that resistant
starch has been found effective against colon cancer by lowering the pH in the colon which
ultimately inhibits the production of injurious by-products of protein fermentation (Koo et al.
2010).
Regarding the carbohydrates content of pseudo cereals, these enclose fewer amounts of mono
and oligosaccharides but rich in starch content. The granules of starch are small in diameter
usually in the range of 1-8 μm with amylose content of about 3-20% (Lindeboom et al. 2005).
2.3.1.2 Non-starch polysaccharides (NSP)

The carbohydrate polymers except starch which make up the outer covering of cereal grains are
termed as Non-Starch Polysaccharides (NSP). As present in bran so a higher extraction rate is
related with an elevated level of NSP (Meuser and Suckow 1986). Nutritionally, NSP are termed
as dietary fiber, these carbohydrate polymers of plant origin which remain intact in human
digestive tract (Bermink 1994). There are two types of dietary fiber which differ on the basis of
their solubility. The components of dietary fiber that are water soluble are called as soluble
dietary fiber and comprises of pectic substances and hydrocolloids while the components that are
insoluble in water are known as insoluble dietary fiber and consist of lignin, cellulose and
hemicellulose. Arabinoxylans (AX) and β-glucans (soluble dietary fiber) are key elements of
cereal grain endosperm such as wheat, barley, oat, rye etc. (Blackwood et al. 2000).
2.3.1.2.1 Arabinoxylans (AX)

All the cereal species possess varied quantity of AX as the highest contents exist in rye (6-8%)
while wheat grain has only 1.5-2.0%. AX is composed of β, 1-4, d –xylopyranosyl subunits as a
linear polymer along with substitution of α -l arabinofuranose at the O-2 and/or O-3-positions
(Izydorczyk and Biliaderis1995). Ferulic acid is a minor element of AX which is attached to
arabinose through an ester linkage at the O-5 position. All the cereals differ substantially in their
AX content, molecular weight and pattern of substitution (Maes and Delcour 2002). AX is
generally classified in two subgroups on the basis of their extraction in aqueous media as a
water-extractable (WEAX) and a water un-extractable fraction (WUAX). Total WEAX in wheat
and rye are 25-30% &15-25%, respectively (Meuser and Suckow 1986).

2.3.1.2.2 β Glucans

β glucans also known as lichenin are the principal non starch polysaccharides in barley (3-7%)
and oats (3.5-5.0%) while > 2% exist in other cereal grains (Lethonen and Aikasalo 1987). The
basic monomers in β glucan are D-glucose chains connected through mixed β,1-3 & β,1-4
glycosidic linkages. β glucan is comparatively flexible with an asymmetrical shape caused by the
two above mentioned different bonds in the polymer (Izydorczyk and Dexter 2008). β glucans
illustrate higher aqueous solubility as compared to AX (38-69% in barley, 65-90% in oats) and
subsequently lead to the formation of viscous solution. Epidemiological studies have depicted
the hypoglycemic and hypocholestrolmic potential as well as immunostimulatory activity of β
glucan via fermentation of colon microflora (Liu 2003). Furthermore, β glucan content of cereal
grains depend upon its genotype and is considered as an essential functional component of food
that can be enhanced by plant biotechnology (McKevith 2004).
2.3.2 Proteins

Protein is regarded as one of the imperative nutrient for the physiological activities of human.
Cereal grains diverge greatly in their protein content due to variation in genotype, agro climatic
conditions etc. Although the proteins are scattered over the whole grain but their quantity differ
within each part of the grain. For example, germ and aleurone layer of wheat grains holds 30%
proteins while endosperm and bran contain ~13 % & ~7%, respectively (Belitz et al. 2009).
Even the protein content varies among all parts of cereal grain but is mostly embedded in the
starchy endosperm. Regarding the amino acid profile of wheat, rye and barley; glutamine is
present in highest concentration (15-31%) followed by proline (12-14%), leucine (7-14%) and
alanine (4-11%) respectively. Essential amino acids such as tryptophan (0.2-1.0%), methionine
(1.3-2.9%), histidine (1.8-2.2%) and lysine (1.4-3.3%) are present only at very low levels and
thus termed as limiting amino acids of cereals but now some new varieties have been developed
via biotechnology, such as high-lysine barley and corn (Shewry and Tatham 1990).

2.3.2.1 Osborne fractions

Conventionally, cereal flour proteins are grouped into four types (albumins, globulins, prolamins
and glutelins) on the basis of their solubility and the process of fractionation as given by
Osborne, 1) albumins are soluble in water, 2) globulins are soluble in dilute salt solutions but
donnot dissolve in water, 3) prolamins are categorized as cereal proteins being soluble in
aqueous alcohols i.e. 60-70% ethanol and 4) glutelins are described as proteins being soluble in
dilute acids or bases. However, it has been found that remarkable fraction of glutelins are
insoluble in dilute acids e.g. acetic acid and the primary structure destroy during extraction in
alkaline media. Currently, a mixture of different solvents including alcohols (50% propanol),
reducing agents (dithiothreitol) and disaggregating compounds (urea) are being employed for its
extraction (Osborne 1907).

Most of the albumins and globulins are considered as metabolic proteins on the basis of their
functionality for example, enzymes/enzyme inhibitors, while oats are an exception that possess
substantial levels of legume like globulins e.g. 12S globulin (Peterson 1978). These metabolic
proteins mostly reside in the aleurone layer, bran and germ while present at very low
concentration in the endosperm. Albumin and globulins are present in lesser concentration than
the storage proteins with an exception of oat globulins that exist about 50% of the total proteins.
Their nutrition profile reveals that amino acid composition of metabolic proteins is rich in lysine
contents. The enzymes included in metabolic proteins are capable to hydrolyse carbohydrates &
proteins and hence fulfil the energy requirements of embryo during germination (Kruger and
Reed 1988).

Along with enzymes, enzyme inhibitors are also a part of metabolic proteins. Many researchers
have segregated and illustrated enzyme inhibitors from germ and endosperm. Usually, the
inhibitors target the hydrolysing enzymes to avert the extensive break down of starch and
proteins during the developmental stages of grain and also protect & defend plant tissues from
animal (insect) or microbial enzymes (Lasztity 1984). Predominant classes are amylase and
protease inhibitors which are concentrated in the albumin/globulin fractions. Amylase inhibitors
can be directed towards both cereal and non-cereal amylases while protease inhibitors directed
towards proteases from both cereals and animals. Some inhibitors appear to be bifunctional
inhibiting amylases as well as proteases (Delcour and Roosney 2010).

Prolamins and glutelins are the primary storage proteins of cereal grains embedded in the starchy
endosperm and provide nitrogen and amino acids to the young seedling during the process of
germination and their concentration vary from 70 to 90% based on total protein content. But
some cereals have an exception such as oat in which prolamins are present in minute quantity
and rice that is devoid of it (Shewry and Tatham 1990). The Osborne fractions of all proteins are
a mixture of complex proteins rather than a single fraction. The prolamin fractions of the diverse
cereals have been classified according to their trivial names such as: wheat (gliadin), rice
(oryzin) (Mandac and Juliano 1978), corn (zein) (Wilson 1991), barley (hordein), oats (avenin)
(Peterson 1978), millet & sorghum (kafirin) (Shull et al. 1991) and rye (secalin) (Gellrich et al.
2005). Similarly, the glutelin fraction of wheat, corn, barley and rye are known as glutenin,
zeanin, hordenin and secalinin, respectively. The prolamin and glutelin fraction of wheat proteins
are collectively known as gluten (Wieser et al. 2006).

Along with its quantitative features, Osborne procedure is also employed in the preparation and
characterization of flour proteins and the enrichment of different protein types. Amino acid
profile, sequence, molecular weight and inter & intra-chain S-S linkages of storage proteins
(prolamins and glutelins) have been broadly explored. These investigations revealed that the
storage proteins of wheat, barley and rye are closely related according to their phylogeny while
oat glutelins are structurally different (Kohler and Wieser 2000). According to two different
principles, storage proteins have been categorized into three groups on the basis of their
structure. Shewry and Tatham (1990) defined all storage proteins as prolamins on the basis of
their molecular weight and sulfur content as high-molecular-weight (HMW), sulfur rich (S-rich)
and sulfur poor (S-poor) prolamins.
The prolamins are also categorized on the basis of their electrophoretic mobility in acid-
polyacrylamide gel electrophoresis (PAGE) with band region termed as a / b (highest mobility),
g (medium mobility) and w (lowest mobility). Their nomenclature is also based on their apparent
size by employing sodiumdodecyl sulfate (SDS- PAGE) e.g. HMW and MW-glutenin subunits
(GS), HMW-secalins, D-, C- and B-hordeins (Shewry and Tatham 1990).

Storage proteins present in rice, maize, sorghum and millet depict similarity among each other
while on the other hand these grains illustrate a significant difference from wheat, barley, oat and
rye. Regarding the amino acid composition of these storage proteins, these possess higher level
of leucine and other hydrophobic amino acids and less quantity of proline and glutamine
(Hamaker et al. 1995). Corn storage proteins, called zeins, can be subgrouped into monomeric
zeins being dissolved in alcohol and cross-linked zeins that dissolve in alcohol with subsequent
heating or breakage of disulfide bonds. On the basis of structure, corn zeins are classified into
four subgroups (Tatham et al. 1985). a-Zeins are the main subgroup comprising of the 71-85% of
total zeins, followed by g-zeins having 10-20% of total zeins and b-zeins & d -zeins with 1-5%
of the total zeins, respectively (Esen 1987). Considering the molecular weight of all these
fraction revealed that a-Zeins have an apparent molecular weight of 19,000Da and 22,000D as
determined by SDS-PAGE while g-zeins have 10,000D to 27,000D MW, b-zeins and d-zeins
possess 18,000D and 10,000D MW, respectively (Wilson 1991).
The prolamins of sorghum and millet are termed as kafirins and on the basis of solubility,
molecular weight and structure these are subdivided into a, b, g and d subgroups (Watterson et
al. 1993). Among all these sub categories, a-kafirins constitutes about 65-85% of the total
kafirins and represents as the main subclass of total kafirins. Other sub groups comprise of > 10
% of total kafirins (Shull et al. 1991). The ratio of prolamins to glutelins (~1:30) is unbalanced in
the storage proteins of rice (Mandac and Juliano 1978). SDS-PAGE patterns of rice prolamins
(oryzins) showed a major band with MW 17,000Da and a minor band with MW 23,000Da.The
apparent MW of glutelin subunit is in a range from 20,000Da to 38,000Da (Juliano 1985).

Protein quantity and quality both are much improved in the pseudocereals grains. The most
limiting amino acid lysine is in higher content in pseudo cereals as compare to the cereal grains.
Arganine and histidine content is high in amaranth and quiona and thus ulitilized in infant
formulas and specialty products for growing children. Bioavailability in terms of protein
efficiency ratio and protein digestibility is the main indicators employed as determinant of
nutritional quality of proteins and pseudo cereals rank higher in this regards. These are low in
prolamins and probably suitable for cealic disease (Drzewiecki et al. 2003).

2.3.3 Lipids

Lipids exist in minute quantity in cereal grains but have a pronounced impact on the functional
attributes of food by conjugating with protein and starch that varies from 1.7-7.0% among
cereals with highest concentration present in germ followed by aleurone layer and endosperm.
Oat is rich in lipid (8%) while wheat grain contains ~2% including essential fatty acids (palmitic
C16:0 & linoleic acid C18:2), phytosterols and fat soluble vitamins (Ruibal-Mendieta et al.
2004). Lipids are classified as free lipids, starch lipids and non-starch lipids (NSL) on the basis
of their extraction under specific conditions (Eliasson and Larsson 1993; Hoseney 1994). NSL
constitute about 75% of the total lipids when extracted with aqueous butanol at normal
temperature.

Fatty acid profile of all cereal grains is similar having 39-69% linoleic acid, 11-36% oleic acid
and 18-28% palmitic acid (Table 2.2). (Delcour and Hoseney 2010). Triglycerides are present in
form of free non starch lipids and free lipids in germ and aleurone layer while phospholipids and
glycolipids are present in the endosperm as bound non starch lipids. Even present in minute
quantity but have a pronounced effect on the baking properties so have been studied
comprehensively. The pseudo cereals are rich in fats content as compare to the cereal grains
with an elevated amount of unsaturated fatty acids. Squalene a bioactive compound present in
pseudo cereals is basically an unsaturated open chain triterpen that is mostly present in the liver
of deep sea fish and other marine species (Lindeboom et al. 2005).
2.3.4 Micronutrients

Minerals and vitamins are two categories of micronutrients which are present mainly in bran,
germ and aleurone layers of cereal grains. The mineral content of cereals ranges from 1.0-2 to
5%. Thus whole flour is rich in mineral as compared to patent flour. Wholegrain cereals have
substantial levels of micro minerals such as iron, zinc and magnesium along with trace minerals
such as selenium. Among cereals, selenium content is highest in rice as 10-13 μg/100g
(Henderson et al. 2003b).
Vitamins are present in the range from below 1 to 50 mg/kg in cereal grains. Mostly, cereal
grains are deficient in vitamin A & C but have substantial content of vitamin B. Thus, cereals are
a good source of B-group and fulfil about 50-60% of the daily requirement of B-vitamins.
Vitamin E is the most important fat soluble vitamin present in the form of the tocopherols in
range of 14-26 mg/kg in cereal grains. Just like minerals, vitamins are also present in the outer
portion of grains hence, milling of cereals into patent flour results in loss of most of the vitamins.
Therefore, whole grain products or products enriched or fortified with micronutrients can be
more beneficial for consumers (McKevith 2004).

Pseudo cereals are also rich source of micro and macro minerals such as calcium, potassium, iron
and zinc. The ratio of calcium and phosphorous (Ca:P) is considered best as 1-1.5 while it is
about 1.9-2.7 in pseudo grains. Vitamins are also present in substantial amount in pseudo cereals
especially the folate content. Buckwheat possess 24.7 mg/100 g total folate while in amanarth it
ranges from 52-70 mg/100g, on the other hand quinoa possess about 10 times more total folate
than spring wheat as 132.7 mg/100g (Schoenlechner et al. 2010).
2.3.5 Bioactive components

These are the biochemical compounds that exist in minute quantities in food matrices and have
health promoting properties. In cereals, these phytochemical are generally present in bran and
germ portion of grains. Phenolic compounds, phytosterols, tocols, dietary fiber, lignins, phytic
acid, cinamic acid, ferulic acid, oryzanols, carotenoids etc. (Liukkonen et al. 2003) are some of
the bioactive components in cereals. Some of these compounds are present in specific cereals
i.e. oryzanols exist in rice bran, saponins are present in oats, β glucans in oat and barley while
alkylresorcinol subsist in rye (Fardet 2010). The factors that may influence the quantity of all
these compounds include nature of grain, cultivars and portion of grain sampled (Adom et al.
2005). Among the phenolic compounds, phenolic acids and flavonoids are mostly present in
whole grains (Heinio et al. 2008) while considering the carotenoids, lutein has been found in
wheat, followed by zeaxanthan and β cryptoxanthin in rice and maize, respectively. The most
promising characteristic of carotenoids is that these are equally distributed in kernel with major
content in endosperm portion (Konopka et al. 2004; Saikia and Deka 2011). Phytic acid and
saponins are usually present in bran portion of cereal grains and termed as antinutritional
components as phytic acid binds the mineral elements there by lowering the bioavailability of
these nutrients (Lopez et al. 2002). Yet it has now been recognized for its antioxidative potential
by chelating the free iron and subsequently suppressing the iron catalyzed oxidative reactions
(Graf et al. 1987). Tocols comprise of tocopherols and tocotrienols and renowned as natural
antioxidants in cereal grains (Nielson and Hansen 2008). β glucan are the prime component of
cell wall of cereals especially oat and barley. In barley these are present in endosperm but in oat
they usually reside in aleurone layers (Peter 2007). These bioactive moieties are renowned as an
effective tool to cure various diseases such as hypercholesterolemia, diabetes, cancer, oxidative
stress, cardiovascular disorders etc. (Kris-Etherton et al. 2002).

2.4 Storage of Cereal Grains

Grain quality is generally determined before harvesting because deterioration may occur during
harvest, drying and storage so the quality of grain can only be improved before harvesting.
Certainly, the keeping quality of kernels may instigate to depreciate in the field before harvest.
Agro-climatic conditions such as rainfall before or during harvesting may promote ear diseases
and early sprouting. Along with it, high moisture content in grains will require more drying
expenses (Bailey 1992). Delayed harvest of wheat will lead to low protein contents, reduced
specific weight and high amylase activity which in turn will acutely lessen the grain quality for
bread making. Inferior quality of crop in the field will give poor quality grain and subsequent
end product. Weeds when present along with the grains may obstruct the harvesting process and
results in high moisture content of grains as well as raise the expenses of cleaning (Evans 2001).

Cereal grains are considered as one of the vital commodities for feeding humanity. Some of the
pre-processing parameters which play a key role in maintaining the end product quality include
drying and storage of grains. Proper storage of grains is basically done to uphold physical,
chemical and biological characteristics of the harvested grain during the storage period (Bailey
1992). The grains can both momentarily be stored on the farm after subsequent harvesting or
depart to the collection centre directly, from where these are transported to bigger storage
facilities. The techniques for sustaining the value of cereal grains are being employed since
prehistoric ages but deterioration is still observed in developed countries (Bell and Armitage
1992).
Storage is related to a wide range of chemical and biological hazards such as sprouting,
microbial spoilage and insects & rodents’ infestation. Appropriate storage is crucial to reduce the
postharvest losses with some of its critical factors such as; moisture content, temperature &
duration of storage (Richard-Molard 2003). Elevated level of moisture in cereal grains during
storage creates a favourable environment for the propagation of mold growth as well as insect
infestation and enhanced rate of respiration. Being a living commodity, when the grains respire
they increase the temperature of storage bin and this may ultimately affect the quality of grains.
Proper ventilation can be an appropriate method by which the excess heat is expelled from the
storage area and ultimately reduced the insect infestation (Reed 2006).

2.4.1 Drying of cereals

Drying is a preliminary step before storage with an aim to eradicate excess moisture from grains
and for the purpose; air has been recognized as an appropriate medium. The parameters of drying
air which must be considered for the development of a suitable drying method includes; drying
temperature, relative humidity, the specific volume and specific enthalpy. The rate of drying
actually depends upon the drying of individual seed (Brook 1992). Generally, the size and
moisture loss represent inverse relation i.e, small grains dried earlier then the large ones.
Similarly, naked kernels also dry easily than those having husk around them. Maize kernel is the
largest among all the cereal grains so it will take more time for drying. On the other hand, when
wheat and rice are considered, wheat grain will dry more easily then the rice kernel owing to the
presence of husk in rice grain (Reed 2006). A wide variety of low to high temperature driers may
be employed for the purpose of drying. High temperature drier removes moisture from the grains
rapidly and destroy insects. These have less time of contact with grains but it can also negatively
influence the properties when not applied properly such as the thermal denaturation of protein.
Natural drying processes have also been exploited such as wind and solar driers e.g., corncobs.
Some of the naturally present modified storage atmosphere comprises of underground storage
and silos blushing with gases such as nitrogen (Delcour and Hoseney 2010).

Cereal grains have moisture content of about 16 to 20% at the time of harvest. This elevated
level of moisture content makes them susceptible to microbial deterioration. So, for suitable
storage the grain must be dried upto the moisture level of 13 to 15%, while when the barley is to
utilize for malt preparation it can be dried to 12%. Drying systems either depend on normal or
high temperature of air as drying medium. The former is a comparatively slow method in which
ambient air is forced through the grain chamber for the purpose of drying (Fan et al. 1976). The
driers having the batch or continuous flow of air use air at the temperature 40 to 120ºC followed
by cool air to remove excess heat and vapours from the grains for appropriate storage. As
moisture and heat are the crucial factors for grain storage so these should be controlled to avoid
any damage to the grains. Low temperature and moisture should be maintained in storage
premises to control microbial infestation. Usually in developing countries crib and bag type
storage is applied for maize cobs at small farms but these are now being substituted by bulk
storage all over the world (Reed 2006).
2.4.2 Methods of storage
2.4.2.1 Crib storage
Crib storage is an obsolete method of grain storage especially ear maize, but still practiced with
proper ventilation and moisture content of 20-25 % for clean grains in moderate climate. When it
has to be applied in warm and humid areas proper fumigation is applied to cribs for control of
insect damage during storage. Suitable aeration is a key factor that can take away 3-5% of
moisture from stored grains and if the method is applied for high moisture grains (> 25%) it
should then be equipped with automatic aeration system (Hall 1957).
2.4.2.2 Bag storage
This type of storage is also applied to small scale level of storage yet has its own importance
such as; easy to carry and transport, each individual lot with specific identity does not require
any specific equipment and can be stacked under suitable shelter. But it requires labour efforts so
can be expensive in areas with high labour cost (Bell and Armitage 1992). Bags made of
different materials such as jute, cotton, hemp etc. are usually utilized for storage but these are
unable to provide protection against deterioration factors. On the other hand, bags prepared from
polypropylene offer better mechanical strength and protect entry of rodents but these may add
extra cost and are also vulnerable to UV radiation deterioration. Although these provide high
mechanical strength but cannot be piled up more than 3m due to the slippery nature of
polypropylene. Jute bags give better opportunity in this regard and can be piled upto 6 m in
storage premises (Bailey 1992).
2.4.3.3 Bulk storage
2.4.3.3.1 Vertical storage
 This type of storage is applied for bulk storage of grains with proficient expulsion from the
concrete silos under the action of gravity. For the proper construction of vertical type silos, an
appropriate layout is the key factor under consideration. The concrete silos or steel bins have
variable holding capacity that depends upon the bulk density of stored grains. Each vertical type
of storage facilities has its own benefits and disadvantages such as the concrete silos have more
durability, large storage capacity, less maintenance and water condensation issue (Fan et al.
1976). However these also have some disadvantages; large setup cost, more manufacture time
then steel bins. While steel bins have flexibility in the fitting of ventilation equipments and
temperature sensors. Both the steel bins and concrete silos vary in the thickness of their walls
such as 150 mm for concrete silo and 5 mm for steel bins, respectively and ultimately differ in
volume with same outside diameter e.g., a 30 m x 5 m steel bin has 12 % more capacity than a
concrete silo. The main drawback of steel bin is rusting, so the bin must be galvanized and all the
nuts and bolts should also be properly painted to avoid appearance of rust spots (Boumas 1985).
2.4.3.3.2 Horizontal storage
Along with vertical type silos and bins, horizontal sheds and ware houses are also an effective
storage method for large scale storage. Although it is quite difficult to pour and remove grains
from these warehouses and need high labour efforts but have low initial construction cost as
compared to the vertical type silos. These sheds also require proper aeration and is usually less
uniform than vertical silos so the air flow is adjusted at higher rate i.e. 0.05 m3/min/ton (Boumas
1985).
2.4.3.3.3 Controlled atmosphere (CA) storage
Basically this type of storage is designed on the basis of variation in the gaseous condition of the
storage equipment. This alteration may be due to the respiration of grains as they are a live
commodity and thus consequently change the level of gases (reduces the O 2 and increases the
CO2). This can also be devised artificially through infusing gases such as nitrogen or carbon
dioxide in the storage premises. This type of storage is observed in conventional underground
cavity storage in which the concentration of oxygen decreases from 0.5 to 0.2% while carbon
dioxide increases from 45-50%, respectively. The change in atmosphere creates an inhibitory
effect on microbial infestation and is usually practiced in some regions especially in Africa
(Busta et al. 1980). Nitrogen generators are used for infusing the gas in sealed premises and this
storage type is mostly employed for long term storage of grains. The moisture content of grains
must be in range of 12-14% for controlled atmosphere storage. Nitrogen gas is lethal for life and
permeation of pure nitrogen can destroy any form of microbes present in the storage facilities.
There is a problem of nitrogen leakage from the premises so it is applied along with oxygen such
as 97% to 99% N2 and 0.5% to 4% O2. The lethal effect of nitrogen depends upon species of
microbes or insects, storage temperature and moisture content. Sometimes carbon dioxide is also
used for CA storage alongside nitrogen (Banks and Sharp 1979).
2.4.3.3.4 Chilled storage
High temperature provides the most favourable environment for the deterioration of grain
quality. Thus, to keep the grains intact, chilled storage is an appropriate method that uses low
temperature conditions and is favourable in cold regions such as tropical areas. For the purpose,
a chiller or a mechanical refrigeration unit is used that may lower the temperature and humidity
ratio of air to the desired level and then enters the chilled air to the storage premises (Sulzer
1993).
2.4.3 Nutritional changes during storage

Nutritional attributes of cereal grains may alter during storage even though these changes will be
very minute. The quality of grains will deteriorate when the storage environment has high
moisture content as it favours the enzymatic activity of grain & microbes consequently leads to
the breakdown of starch (Macrae et al. 1993). The presence of favourable conditions such as
elevated temperature and moisture activates the lipolysis of unsaturated fatty acid and thus
causes rancidity in grains. However, a little or no change occurs in protein and micronutrient
content of cereal grains even if stored for up to 6 months in disinfected conditions. On the other
hand, proper storage is also necessary to enhance the milling yield and cooking characteristic of
rice grains as their expansion ratio increase during cooking after appropriate storage. To improve
the cooking characteristics and eating quality of rice grains they should be stored for 3 to 4
months (Pomeranz 1984).