Exercise 25
Aim: Observation and comment on the setup.
A. Anaerobic Respiration
Principle: Breakdown of food substances to yield energy in the absence of oxygen is
called anaerobic respiration. It is observed in several soil anaerobic microorganisms, yeast
and certain types of tissues in human body. Anaerobic respiration yields much less energy
per mole of glucose as compared with aerobic respiration. In germinating seeds/flower
buds the equation for anaerobic respiration is C6H12O6 alcohol CO2+C2H5OH.
dehydrogenase
Requirement: Germinating seeds (gram/urad/moong), flower buds, a small test tube/
glass vial, petridish, a plastic tray slightly bigger than the size of petridish, mercury, forceps,
KOH pellets, burrette stand with clamp.
Procedure
• Take a test tube and
completely fill it with mercury.
Invert it over a petridish which
is also filled with mercury. Seeds
There must be a continuous
column of mercury in the
test tube.
• Tilt the test tube slightly and
with the help of forceps
Mercury
introduce 3 - 4 healthy
germinating gram seeds.
• Gently tap the test tube with
your finger nail/forceps so
that the seeds move upwards
Fig. 25.1 Experimental setup for
in the mercury column. the study of rate of
• With a clamp fix the test tube anaerobic respiration
to a stand and keep the setup
undisturbed for two hours.
• Observe the setup.
• Introduce 3-4 KOH pellets in
the same way as seeds were
introduced. Observe the changes.
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Observation
A space is formed at the top of the test tube due to downward displacement
of mercury. When KOH pellets are introduced, the gap slowly disappears
and mercury again fills up the entire tube.
Discussion
The germinating seeds respire in a situation when these are completely cut
off from air in the presence of a continuous column of mercury. The carbon
dioxide gas released gets collected at the top of the tube and displaces
mercury. The CO2 released disolves in KOH and the mercury level rises again.
This establishes the fact that seed/buds have respired anaerobically.
B. Phototropism
Principle: Light is responsible for inducing many responses and physiological processes in
plants. Light induced growth response in plants is called phototropism. Shoot is positively
phototropic. Light controls the distribution of auxins in the shoot tip and a greater quantity of
auxin accumulation occurs in the shaded portion leading to rapid cell division resulting in faster
growth and the bending or curvature of the stem towards light.
Requirement: Two herbaceous potted plant of same species, or seeds of mustard, an earthen
pot filled with sandy soil, water.
Procedure
• Keep one potted herbaceous plant on a table near a window
for a week and the other in the open space (Fig. 25.2). Water
the plant, regularly and observe.
OR
• Germinate about 20-30 mustard seeds each in two pots.
Keep one pot on a table near a window and the other in
exposed open space. Water them daily. Observe the
curvature of seedlings after a week in both setups. Fig. 25.2 A potted plant
showing phototropism
Observation
You will observe that the pots with plant/seed kept near window exhibit a
distinct bending towards the light.
Discussion
The curvature or bending of shoot towards the source of light is due to the
redistribution of auxins in the apical meristem of plants kept near the
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Exercise 25
window. The shoot/seedling kept in open space do not show any curvature.
Shoots always exhibit positive phototropism.
C. Apical bud removal (Apical dominance)
Principle: Stem grows due to divisions of the apical meristematic cells situated at the tip of
stem. Plants possess axillary (lateral) buds in the axil of leaves. The axillary buds are generally
dormant and are not expressed because the apical bud imposes a natural inhibition over the
growth of lateral buds. This influence exerted by the apical bud over the axillary buds is called
apical dominance.
Requirement: Two potted plants of same species with sparse branching- (Chenopodium/Croton/
Tecoma/Petunia/Hibiscus), knife/scalpel/blade.
Procedure
• Using a blade cut the tip of the stem of the experimental plant (plant
A). Keep another plant of the same species without cutting its tip as
control (plant B).
• Water both the plants regularly and observe.
Observation
After about 7-10 days, several lateral (axillary) branches will be seen in plant
A as compared to plant B.
Discussion
When the apical bud is removed in plant A, the apical dominance no longer
exists, as a result the lateral buds are activated and give rise to lateral
branches.
D. Suction due to transpiration (Transpiration pull)
Principle: Transpiration refers to loss of water in vapour form from the aerial parts of
plants. Transpiration develops a pull/upward suction force because of the presence of a
continuous water column (formed due to cohesive and adhesive properties) that starts
from the mesophyll cells of leaves (from where water is lost) to the root hairs (from where
water enters the plants). This is called transpiration pull. This force is responsible for the
absorption and upward conduction of water and minerals in the stem.
Requirement: A healthy branch of a shrub or a tree (20-25 cm long), beaker (100mL) or a
glass tube (15 cm long and 0.5 cm diameter), a rubber tube (8-10 cm long and 0.5 cm diameter),
50ml beaker, mercury, scalpel/knife, grease or petroleum jelly, tray, stand with clamp.
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Procedure
• Insert a graduated 1 mL pipette to one
end of a rubber tube so that it fits tightly.
• Select a twig of appropriate diameter
and size and make an oblique cut with
a knife taking care not to rip off the bark.
Moisten the cut edge with water.
• Fill the rubber tube and pipette with
water by suction.
• Insert the twig into the other end of the
rubber tube and tie a thread to make it
air tight (Fig. 25.3).
• Fix the set up with the help of burette Fig. 25.3 Experimental setup showing
stand and clamp as shown in the suction due to transpiration
Fig. 25.3.
• Take a glass tube/beaker and fill it with coloured water (using
eosin/methylene blue/blue ink) and keep the nozzle of the pipette
in it.
• Note the level of coloured water in pipette. Keep the setup in
sunlight/under a fan for an hour and note the final level of
coloured water in the pipette.
Observation
Initially when the experiment is set up, a continuous column of water is
observed in the pipette. After an hour or so coloured water rises in the
pipette.
Discussion
A strong suction force was developed in the water column due to transpiration
from leaves. First a water potential gradient was created between the stomata
and the spongy mesophyll. As a result of transpiration through stomata,
water from the mesophyll moved into the stomata. A similar water potential
gradient was built up sequentially from mesophyll cells - leaf veins- leaf
midrib xylem - petiole xylem - stem xylem - water column in the experiment.
The intense suction pressure generated sucked water against gravity.
In nature this suction force is transmitted into the root xylem and root
hair, which enables plants to absorb water from the soil.
Questions
1. What other material could be used for demonstrating anaerobic respiration?
2. Would the same plant kept near the window respond similarly to artificial light?
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Exercise 26
Aim: To study the enzymatic action of salivary amylase on starch.
Principle: An enzyme is a biological catalyst. The enzyme salivary amylase or ptyalin present
in saliva initiates the breakdown of starch, which hydrolyzes it into disaccharide maltose,
isomaltose and short chain glucose polymers called - dextrins. Enzymatic activity of salivary
amylase can be tested by detecting the presence or absence of starch.
Requirement: Glass wares: Test tubes, cavity blocks, beakers, dropper, and funnel; Chemicals:
NaCl, Na2HPO4, KH2PO4, Iodine crystals, potassium iodide; Equipments: water bath or
incubator thermometer; Miscellaneous - cotton, rubber, distilled water, test-tube stand, test-
tube holder.
Preparation of reagents
(i) 1% starch solution: Add 1 g of soluble starch to 10ml of distilled water and mix them. Boil
90 ml of distilled water and to it add 10 mL of starch solution already prepared by stirring.
Leave the solution overnight and then filter to get 1% starch solution.
(ii) 1% NaCl solution: Dissolve 1 g of NaCl in 100 mL of distilled water.
(iii) Iodine solution (Lugol's): Dissolve 1 g of iodine crystals and 2g of potassium iodide in
100 mL of distilled water.
(iv) Preparation of buffer solution at pH 6.8: Buffer solution can be prepared by dissolving one
buffer tablet of 6.8 or 7 pH in 100 ml distilled water or prepare M/15 Na2HPO4 solution
(9.67g Na2HPO4 in 1000 ml of distilled water) and M/15 KH2PO4 solution (9.06g of
KH2PO4 dissolved in 1000 ml of distilled water). Mix equal volume as per requirement to
get buffer solution.
Procedure
• Take cotton soaked in distilled water. Remove the excess water
by pressing and then spread the moistened cotton over the mouth
of a funnel in such a way that it acts as a filter. After cleaning
mouth, chew a piece of rubber/cotton and pour the saliva into
the funnel. Saliva filtered through wet cotton will be collected in
the test tube. Avoid using filter paper for filtering saliva. Take
1 ml of saliva and add 19 ml of distilled water to get saliva solution.
• Take two sets of test tubes (8-10 test tubes in each set) in two
separate test tube stand each containing 1 mL of iodine solution to
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act as indicator tubes. Mark them 1, 2, 3 . . .. in both test tube
stands.
• Switch on the electric water bath or oven. Set the temperature at
370C. Maintain uniform temperature (370C) of the water in water
bath or water in a beaker inside incubator throughout the
experiment.
• In a test tube, take 10 ml of starch solution, 2 ml of 1% NaCl
solution and 2 ml of buffer solution. Mix them well and transfer
half of the solution into another test tube. Mark one test tube as
experimental tube and the other as a control tube.
• Now transfer both experimental and control test tubes to the water
bath or keep them in the beaker containing water inside incubator
for about 10 minutes so that temperature of solutions reaches
370C.
• Add 1 ml of saliva solution to the experimental tube and 1ml of
distilled water to control tube. Keep both tubes in water bath/
incubator throughout experiment.
• With the help of a dropper, take a drop each from experimental
and control tubes and pour it into two separate indicator tubes
(marked 1) containing iodine (from two series of indicator tubes -
one for experimental and other for control). Record the time of mixing
as zero minute reading and note the change in colour of iodine in
both tubes.
• After two minutes, again transfer a drop each from experimental
and control tubes to indicator tubes (marked 2) and note the
colour of iodine. Repeat the step at interval of every two minutes
till the colour of iodine solution does not change any further
(achromatic point). Always take the same amount of solution
throughout the experiment to be added to iodine tubes.
• Compare the series of experimental tubes with the control iodine
tubes.
Time (Min) Control Indicator tube Experimental
Indicator tube
0 Blue colour Blue colour
2 ____________ ____________
4 ____________ ____________
6 ____________ ____________
___ ____________ ____________
___ ____________ ____________
___ ____________ ____________
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Exercise 26
Observation
(i) Note the colour in control indicator tubes.
(ii) Note the colour change in both test tubes and the time taken for the
change.
(iii) Perform the Benedict's test for confirmation of the presence of
reducing sugar.
Discussion
On the basis of following questions draw your conclusion:
• Did the colour change occur in both sets?
• Which set showed colour change and why?
• Which set did not show colour change and why?
• Which set showed positive Benedict’s reaction and what does it confirm?
• How much time did it take to reach the achromatic point (no change in
the colour of the indicator)?
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Exercise 27
Aim: To study the effect of temperature on the activity of salivary amylase.
Principle: Most of the enzymes are proteinaceous in nature. Over a limited range of
temperatures, the rate of enzyme catalyzed reactions increases as temperature rises. The rate
of many biologic reactions roughly doubles with a 100C rise in temperature and is halved if the
temperature is decreased by 100C. There is an optimal temperature at which the reaction is
most rapid. Above and below this, the reaction rate decreases sharply.
Requirement: Glass wares: test tubes, cavity block, beakers, dropper, funnel, test-tube stand,
test-tube holder; Chemicals: NaCl, Na2HPO4, KH2PO4, Iodine crystals, potassium iodide,
Prepared Reagents as in Experiment 26; Equipments: water bath or incubator,
thermometers; Miscellaneous - cotton, rubber, distilled water,
Procedure
The first four initial steps are the same as in previous Experiment 26.
• In a beaker, take 15 mL of starch solution, 3 mL of 1% NaCl
solution and 3 mL of buffer solution. Mix them thoroughly. Divide
the solution into three test tubes and mark them as A, B and C.
All three test tubes are experimental tubes.
• Now transfer experimental tube A into a beaker containing ice
and a thermometer for recording temperature. Temperature of
this beaker is to be maintained at 50C. Transfer the second
experimental tube B into water bath set at 370C and the third
experimental tube (C) into the beaker maintained at 700C.
• Without taking them out add to the three experimental tubes
(A, B,and C), 1 mL of saliva solution.
• Take a drop from each of the experimental tubes with the help of
a dropper and add it to the corresponding indicator tubes
containing iodine solution. Note this time as zero minute reading.
• At intervals of every 2 minutes keep on repeating the above step
and note the change in colour of iodine solution. Continue this
till the colour of iodine does not change.
• Note the time taken for different experimental tubes till they do
not give any colour with iodine.
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Observation
To digest completely 5 mL of 1% starch solution, 1 mL of diluted enzyme
takes ------- minutes at 50C, ------- minutes at 370C and -----minutes at
700C.
Time minute Reaction with iodine Reaction with iodine Reaction with iodine
from experimental tube A from experimental tube B from experimental tube C
0 Blue colour Blue colour Blue colour
2
4
6
8
10
12
____
____
____
Discussion
On the basis of the following questions draw your conclusion:
• At which temperature the reaction is optimum?
• Did all three sets of tubes reach achromatic point? If not, why so?
• What inference do you draw about enzyme activity from your experiment?
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