Practical Exercise No. Notes. V.P.S.

Shekhawat
Cell inclusions: Starch grains (Potato and Rice); Aleurone Layer (Maize)

Aim:
Study of various cell inclusions
Theory:
The stored energy in plant tissues is preserved in the form of form of carbohydrates, lipids and
proteins. Plant cells deposit storage resources of carbohydrates, proteins, and neutral lipids in
subcellular particles termed starch granules, protein bodies, and oil bodies, respectively.

Starch is abundant in all major agricultural crops. Although it is the major energy reserve for a
large variety of higher plants such as cereals, legumes and roots and tubers, over 90% of world
starch production comes from cereals. Maize is by far the most important source of starch in the
world contributing to over 80% of world starch production, while wheat (>8%), potato (>5%),
and cassava (>5%) mainly share the rest of the production. Starches such as rice, barley, oats,
sweet potato, sago, etc., also contribute to the world starch production in small quantities.

Native starches have diverse properties and have been utilized for various food (54%) and
nonfood (46%) applications. Physical and chemical modifications have been employed to
improve the properties of native starches and make starch derivatives, and therefore widen the
scope of their applications.

Starch and starch derivatives have a growing demand in the industry because it is a renewable
and biodegradable resource, abundant, environmentally friendly, cost competitive, and versatile.
Storage starch is deposited in the endosperm (in amyloplasts) of the seed, while transient starch
is deposited in leaves (in chloroplasts). Starch is deposited as semicrystalline granules and
consists of two main glucan polymers named amylose and amylopectin. Amylose is
essentially a linear polymer with - (1,4) linkages between glucose units, amylopectin is highly
branched polyglucan. In amylopectin molecules, linear chains connecting glucose monomers via
-(1,4)-linkages, are interconnected via -(1,6) linkages. The starch granule is mainly (98- 99%)
composed of amylose and amylopectin, as discussed above. Although, the ratio of amylose to
amylopectin in a normal starch is about 1:3, the exact value depends on the botanical origin.

Starch grains consists of hilum around which dense and watery starch layers are arranged. The
position of the hilum may be concentric or eccentric. The shape of the hilum may be pointed or
fissured. The aggregation of the Starch grains may be; simple, compound or semicompound.

Depending on the biological origin, starch granules demonstrate different shapes, sizes, and
morphology increasing its specificity in food applications. Variously shaped granules such as
lenticular/oval, round/spherical, polygonal, disk, truncated ellipsoid, and irregular are found in
nature with size ranging from 1 to 100 m in diameter. Granules from roots and tuber starches
are relatively larger in size (2-100 m) and have oval shape although some varieties with round,
polygonal, and irregularly shape granules are found.



(A) Potato Starch
Obtained from the tubers of Solanum tuberosum, family Solanaceae. The granules are flatten
ovoid or sub spherical up to 100m. Few granules are compound, with 2 or 3 components. The
hilum is a point at the narrow end of the granules and concentric striations are well marked. It is
used as a disintegrant in making tablets.

Requirement:


1. Potato tubes
2. Slides
3. Cover Slips
4. Beakers
5. Muslin Cloth
6. Iodine Solution
7. Glycerine

Procedure:
1. Peal a raw potato and cut into small pieces
2. Grind them in a motor and pestle with sufficient water.
3. Collect the potato homogenate into a beaker and add enough water.
4. Then filter the homogenate through a muslin cloth to remove the particles.
5. Allow the filtrate to settle. Starch rapidly settles at the bottom. Decant the starch free
supernatant carefully.
6. Wash 3-4 times and decant the supernatant. Collect the compact mass of starch and allow
it to dry.
7. Observe the isolated starch

(B) Rice Starch
It is isolated from rice grains (From the fruits of Oryza sativa family-Poaceae). Individual
granules of rice starch measure from 3-5 to86 m in diameter. They are polyhedral with sharp
angles without concentric striations; a hilum is visible in the larger granules. Compound
granules are present and consist of from 2-150 components.

Requirements:
1. Rice Grains
2. Slides
3. Cover Slips
4. Beakers

5. Muslin Cloth
6. Iodine Solution
7. Glycerine

Isolation Procedure
1. Take 100g of broken pieces rice and wash them to remove the impurities.
2. Now crush them in powder form and soak them in sufficient amount of 0.4% NaOH
solution for 20 minutes to soften them.
3. This ground mixture is mixed with water for 20 minutes.
4. The mixture is then stirred on a magnetic stirrer for 5 mins
5. Filter it using a muslin cloth
6. The remaining filtrate is allowed to stand in a beaker for some time.
7. The starch will settle down and will be separated and dried and identification tests can be
now performed on this starch.

(C). Maize Starch

It is prepared from grains of fruit of Zea mays family Poaceae. Maize Starch consists of grains
that are nearly uniform in size Consists of granule that are fairly uniform in size measuring 5-25
m, they are polyhedral with blunt angles or more or less rounded. In the center there is often a
small cleft, or two or three radiatingfrom the center, the position of the hilum. The granules are
mostly simple.

Isolation Procedure
1. Maize grains are first washed with running water to get rid of dust particles and adhered
organic matters.
2. They are now softened by soaking in water for 12 to 24 hrs
3. Crush them to separate the embryo and the epicarp.
4. Add distilled water and filter it using a muslin cloth
5. The milky slurry thus obtained is a mixture of starch and gluten particles which is then
subjected to centrifugation
6. Thus, the starch which being relatively heavier settles at the bottom and the gluten being
lighter floats on the surface and removed quickly by a jet of water.






















Observations:
Macroscopic Characteristics:
S.No. Characters Description
Potato Rice Maize
1. Form Fine Powder In masses Fine Powder
2. Color Slightly Yellowish White White
3. Odour None None None
4. Taste Mucilaginous Mucilaginous Mucilaginous
5. Solubility Gives colloidal
solution when boiled
with water
Gives colloidal
solution when boiled
with water
Gives colloidal
solution when
boiled with
water
6. pH Neutral/Faint Acidic Alkaline Neutral

Microscopic Characteristics:
S.No
.
Characters Description
Potato Rice Maize
1. Nature Mostly Simple, but
occasionally compound
Simple and
Compound
Simple
2. Shape Subspherical or ovoid
and often slightly
flattened
Polyhedral with
sharp angles
Polyhedral or
rounded
3. Size Subspherical grains are
upto 35 micrometer in
length and ovoid grain
vary from 30 to 100
micrometer
4-10 micro meter but
average grains are
about 6 micro meter
compound grains
may contain upto 150
components and may
measure upto 30
micrometer in width
and 20 micrometer in
length
5-30 micro meter
Majority with 12-18
micro meter
4. Hilum and
Striations
The hilum occurs as a
small point towards the
narrower end of the
grain and is surrounded
by well marked
concentric striations
No striations are
visible and only very
occasionally a hilum
can be distinguished
as a small central
dot.
The grain exhibit no
concentric striations,
but a hilum can
usually be seen either
as a central point or
as a 2-5 rayed
fissure.

Observation and result:


Aleurone Layer (Maize)

The mature corn kernel is composed of four principle parts: tip cap, 0.8%; pericarp, 5.3%; germ,
11.9%; and endosperm, 82% . Immediately beneath the testa is the first layer of the endosperm,
the aleurone, a single layer of highly proteinaceous cells having thick, tough cell walls. Aleurone
comprises about 3% of the weight of the kernel. Although structurally part of the endosperm, the
aleurone, along with the pericarp and tip cap, is included in the fiber fraction.

(1) Cut thin sections transverse and longitudinal sections of the grain
(2) Place a section in a little Millon's reagent ; if the protein contents do not turn red quickly,
warm the slide.
(3) Note that the greater portion of the endosperm consists of cells packed with starch-grains, but
the outer- most layer (" aleurone " layer) consists of cubical cells containing protein grains.
Protein Grains ("Aleurone" Grains) are found in various parts where food is stored, but are
especially abundant and large in seeds.