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Laboratory Manual

Biology
Class XII


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FOREWORD
The National Council of Educational Research and Training (NCERT) is the
apex body concerning all aspects of refinement of School Education. It has
recently developed textual material in Biology for Higher Secondary stage
which is based on the National Curriculum Framework (NCF)2005. The NCF
recommends that childrens experience in school education must be linked
to the life outside school so that learning experience is joyful and fills the gap
between the experience at home and in community. It recommends to diffuse
the sharp boundaries between different subjects and discourages rote learning.
The recent development of syllabi and textual material is an attempt to
implement this basic idea. The present Laboratory Manual will be
complementary to the textbook of Biology for Class XII. It is in continuation
to the NCERTs efforts to improve upon comprehension of concepts and
practical skills among students. The purpose of this manual is not only to
convey the approach and philosophy of the practical course to students and
teachers but to provide them appropriate guidance for carrying out
experiments in the laboratory. The manual is supposed to encourage children
to reflect on their own learning and to pursue further activities and questions.
Of course the success of this effort also depends on the initiatives to be taken
by the principals and teachers to encourage children to carry out experiments
in the laboratory and develop their thinking and nurture creativity.
The methods adopted for performing the practicals and their
evaluation will determine how effective this practical book will prove
to make the childrens life at school a happy experience, rather than
a source of stress and boredom. The practical book attempts to provide
space to opportunities for contemplation and wondering, discussion
in small groups, and activities requiring hands-on experience. It is
hoped that the material provided in this manual will help students
in carrying out laboratory work effectively and will encourage teachers
to introduce some open-ended experiments at the school level.
YASH PAL
Professor and Chairperson
National Steering Committee
National Council of Educational
Research and Training
New Delhi
21 May 2008


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PREFACE
The development of the present laboratory manual is in continuation to
the NCERTs efforts to improve upon comprehension of concepts and
practical skills among the students. The present laboratory manual will
be complementary to the textbook of Biology for Class XII.
The expansion of scientific knowledge and consequently the change
in the system of education has led to the development of new methods of
instructions. Today the stress is laid on the enquiry approach and
discussion method instead of lecture method of teaching. Biology is now
something more than observation of living organisms. Study of Biology
includes microscopic observations to reveal minute internal details of the
organism, biochemical testing to understand complex reactions taking
place inside the organisms, experiments with live organism to understand
various physiological processes and even much more. In other words
experiments in Biology truly represents an interdisciplinary approach of
learning.
The new syllabus of Biology has been designed to cater to the needs of
pupil who are desirous of pursuing science further. The fundamental
objective of this course is to develop scientific attitude and desired
laboratory skills required for pursuing Biology as a discipline at this level.
A similar approach has been taken while formulating the practical syllabus
of Biology for higher secondary stage. The practical syllabus includes
content based experiments, which help in comprehension of the concepts.
There are altogether twenty-five exercises in the present manual which
are based on Biology curriculum for Class XII. For each practical work,
principle, requirements, procedure, precautions, observations, discussion
and the questions are given in the book. The methodology of preparation
of any reagent, if required, has been given alongwith the requirements,
for the convenience of students and teachers. The questions are aimed to
develop learners understanding of the related problems. However, teacher
may provide help in case the problem is found to be beyond the capability
of the learner. Precautions must be well understood by the learners before
proceeding with the experiments and projects. In addition to the core
experiments enlisted in the syllabus for Class XII emphasis has also been
given for pursuing Investigation Project Work. It is expected that these
informations will motivate the students to take up independent project
work on topics of their interest.
Appropriate appendices related to the observation and study of
organisms are given along with the experiment. International symbols for
units, hazards and hazard warnings are given at appropriate places in
the book. It is expected that this will make the learners more careful about
the environment and make them careful while dealing with the equipments
and chemicals in the laboratory.


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vi
It gives me a pleasure to express my thanks to all those who have been
associated at various stages of development of this laboratory manual. It is
hoped that this practical book will improve teaching-learning process in
Biology to a great extent. The learners will be able to understand the subject
well and will be able to apply the acquired knowledge in new situations.
I acknowledge with thanks the dedicated efforts and valuable contribution
of Dr Dinesh Kumar, coordinator of this programme and other team members
who contributed and finalised the manuscript. I especially thank Professor
G. Ravindra, Director (Incharge), NCERT for his administrative support and
keen interest in the development of this laboratory manual. I am also grateful
to the participating teachers and subject experts who participated in the
review workshop and provided their comments and suggestions which helped
in the refinement of this manual. We warmly welcome comments and
suggestions from our readers for further improvement of this manual.
HUKUM SINGH
Professor and Head
Department of Education in
Science and Mathematics


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MEMBERS
Animesh K. Mohapatra, Associate Professor, Regional Institute of Education,
NCERT, Ajmer
B.K. Tripathi, Professor, DESM, NCERT, New Delhi
C.V. Shimray, Assistant Professor, DESM, NCERT, New Delhi
N.V.S.R.K. Prasad, Associate Professor in Botany, Sri Venkateshwara College,
New Delhi
P.K. Durani, Professor (Retired), DESM, NCERT, New Delhi
Sunita L. Varte, Assistant Professor, DESM, NCERT, New Delhi
S.P. Sinha, Professor of Zoology (Retired), TM Bhagalpur University, Bhagalpur
V.V. Anand, Associate Professor , Regional Institute of Education, Mysore
MEMBER-COORDINATOR
Dinesh Kumar, Associate Professor, DESM, NCERT, New Delhi
LABORATORY MANUAL DEVELOPMENT TEAM


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The National Council of Educational Research and Training (NCERT) acknowledges the valuable
contribution of the individuals and organisations involved in the development of this laboratory
manual. The council also acknowledges the valuable contribution of the following academics for
reviewing and refining the manuscript of the laboratory manual: A.K. Sharma, Reader in Zoology,
University of Lucknow, Lucknow; Iswant Kaur, D.M. School, RIE, Bhopal; K. Muralidhar,
Professor, Department of Zoology, University of Delhi, Delhi; K.K. Sharma, Professor Department
of Zoology, M.D.S. University, Ajmer; M.M. Chaturvedi, Professor Department of Zoology,
University of Delhi, Delhi; Nazir Ahmad Kakpori, Department of Education, Govt of J ammu &
Kashmir, Srinagar; Reena Mohapatra, St. Stephens Senior Secondary School, Ajmer; Savita
Sharma, Mount Carmel School, Dwarka, New Delhi; Savithri Singh, Professor and Principal,
Acharya Narendra Dev College, New Delhi; Shalu Dhawan, Amity International School, Saket,
New Delhi; Shivani Goswami, Mothers International School, New Delhi; V.K. Srivastava, Reader
in Zoology, J .N. College, Pasighat; Vijay Kumar, Delhi State Science Teacher Forum, New Delhi.
We also acknowledge the contributions of Anil Kumar and Binita Kumari, J unior Project
Fellows, DESM, NCERT, New Delhi.
Special thanks are also due to Hukum Singh, Professor and Head, DESM, NCERT for his
interest in the work and administrative support.
The Council also acknowledges the efforts of Surender Kumar, Narender Kumar Verma, Monika
Rajput and Girish Goyal, DTP Operators, for helping in shaping this laboratory manual. The
contributions of Publication Department of NCERT in printing out this laboratory manual are
also duly acknowledged.
ACKNOWLEDGEMENT


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FOREWORD
PREFACE
Introduction
Exercise 1 : To study the reproductive parts of commonly
available flowers
Exercise 2 : To calculate percentage of pollen germination
Exercise 3 : To study pollen tube growth on stigma
Exercise 4 : To study the discrete stages of gametogenesis in
mammalian testis and ovary
Exercise 5 : To study and identify various stages of female
gametophyte development in the ovary of a flower
Exercise 6 : Preparation and study of mitosis in onion root
tips
Exercise 7 : Study of stages of meiosis using permanent slides
Exercise 8 : To study the blastula stage of embryonic
development in mammals, with the help of
permanent slide, chart, model or photograph
Exercise 9 : To verify Mendel's Law of Segregation
Exercise 10 : To verify the Mendels Law of I ndependent
Assortment
Exercise 11 : Preparation and analysis of Pedigree Charts
Exercise 12 : To perform emasculation, bagging and tagging
for controlled pollination
1
5
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CONTENTS
iii
v


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x
Exercise 13 : Staining of nucleic acid by acetocarmine
Exercise 14 : To identify common disease-causing organisms
and the symptoms of the diseases
Exercise 15 : To study the texture of soil samples
Exercise 16 : To determine the water-holding capacity of soils
Exercise 17 : To study the ecological adaptations in plants living
in xeric and hydric conditions
Exercise 18 : To study the adaptations in animals living in xeric
and hydric conditions
Exercise 19 : To determine the pH of different water and soil
samples
Exercise 20 : To study turbidity of water samples
Exercise 21 : To analyse living organisms in water samples
Exercise 22 : To determi ne the amount of Suspended
Particulate Matter (SPM) in air at different sites in
a city
Exercise 23 : To study plant population density by quadrat
method
Exercise 24 : To study plant population frequency by quadrat
method
Exercise 25 : Study of homologous and analogous organs in
plants and animals
Investigatory Project Work
Project 1 : To study the effect of pH on seed germination
Project 2 : Quantitative analysis of phytoplankton in a water
body
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Laboratory is a place where ideas and concepts can be tested through
experiments. Biology, like any other discipline of science, is based on
experimental work and therefore practical forms an integral part of learning.
Biology laboratory provides a unique learning environment where learners
inculcate scientific temper, develop relevant skills and get exposed to realms
of techniques and methodologies of scientific investigations. Laboratory
investigations in Biology increase the reasoning abilities, bring scientific
attitude in a learner and also help in acquisition of skills of scientific processes.
Also, observation of nature and the living organisms found in it is no less
important for the understanding of many aspects of the subject especially
the diversity of the living organisms, their systematic study, their relationships
among themselves and with the environment. Knowledge in the field of
Biology can be acquired or constructed only on the basis of correct
observations and experimentally verifiable processes.
Biology laboratory thus provides the learners an environment where the
process of learning is facilitated by hands-on experiments. Biology is a
unique discipline in the sense that it does not merely deal with the study of
morphology, anatomy, physiology and reproduction of the living organisms,
rather, understanding of the subject requires understanding of a number of
interdisciplinary areas and approaches. On one hand, a biologist needs to
be sufficiently skilled in handling the enormous diversity of the living
organisms, be it plants, animals, fungi or even microscopic bacteria, while
on the other hand, a biologist should be able to understand the biochemical,
molecular, physiological, behavioural, genetic and many other phenomena
pertaining to the living organisms. The study of intricate relationship of
different types of organisms among themselves and also with its environment
is an important concern of a biologist. Thus, experiments and exercises in
Biology train a learner about skills of observations, manipulation of the
organisms for the study of internal details, biochemical as well as molecular
composition and processes, investigation of the abiotic environment and even
analysis of phenomena like inheritance and evolution.
As far as the study of the living organism is concerned, correctness of the
method is very important. Such a study may be very simple, e.g., study of
habit, habitat and external features of the plants or animals, or, it may involve
certain manipulations like dissection and section cutting of the parts of the
organisms to study the minute details. Very often observation and study of
the magnified image of the minute parts under a microscope provides a
better insight about the features of the organisms. However, microscopic
study involves certain specific skills depending on type of the organisms/
tissues/ cells to be studied. It involves specific preparations (peeling, section
Introduction


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2
LABORATORY MANUAL: BIOLOGY
cutting, fixation, staining, dehydration, mounting, etc.) so that microscopic
examination reveals the expected details. As histological and cytological
observations give us only static pictures of the continuous processes, analysis
of biochemical, physiological and ecological aspects need certain other kinds
of skills such as preparation of chemicals and reagents, designing and
performing an experiment, observation and recording of data and ultimately
interpretation and drawing conclusions. While performing experiments,
honesty in recording of data and its correct presentation is very important
as it is not only useful in the logical interpretation but also helps in the
identification of errors.
In order to perform experiments successfully, a learner needs to go to
the Biology laboratory well prepared. This includes the following:
1. Laboratory Record Book: For maintaining all the information including
recording of data and its interpretation.
2. Dissection Box: A dissection box is required in the Biology laboratory
for various purposes like handling and manipulation of living materials,
performing experiments, preparation of slide, etc. A dissection box
should contain scissors (two pairs, one small with fine tip and one
larger), scalpels (one small and one medium sized), forceps (two, one
small with sharp fine tips and the other medium sized with blunt tips),
dissecting needles (two), razor, hand lens, dropper, fine brush, etc.
3. Laboratory Manual
4. A Laboratory Coat or Apron
5. A Hand Towel
While in the laboratory a student should be very careful and methodical.
One should listen carefully to the instructions given by the teacher/
instructor before performing an experiment. In the biology laboratory a
student has to handle a number of sharp objects and hence necessary
precaution and care should always be taken while handling objects like
scissors, forceps, needles, scalpel, razor, etc. It is also very important to
follow the safety instructions mentioned on the instruments and/ or on the
label of the reagent/ chemical. Students should also be aware about the
use of the First-aid Box so that in case of any accident or injury the
preliminary aid can be provided to the affected person.
While describing the experiment students are expected to follow a pattern
in which the aim of the experiment, its principle, list of the materials to be
used, procedure, observation table (if required), inference and discussion
should be given. Necessary precautions to be taken should also be
mentioned appropriately in the procedure or at the end. There are a few
experiments in which field visit is essentially required. For this all the
necessary preparations (materials, equipments, reagents and chemicals)


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3
EXERCISE 1
should be made in advance. Drawing of illustrations is also an important
component of the practical in Biology. Students are expected to follow certain
fundamental rules while drawing the illustrations so that it reflects your
observations correctly.
Make your illustrations using pencil only and always use white drawing
sheet. Illustration should be in the centre of the page.
Drawing of an object (plant, animal or experimental set-up) should be
proportionate in size.
Draw your illustrations keeping the object before you.
Drawing must be clear with simple outlines.
Appropriately label your drawing. Parts of the drawing should be
indicated by straight horizontal line or arrow. Two lines or arrow should
never cross each other. As far as possible, labelling should be done on
the right side of the drawing. An appropriate legend or heading of the
drawing should also be given below it.
About the Manual
The main objective of the manual is to introduce the students of higher
secondary stage to the fascinating world of plants, animals and microbes
and their complex biological phenomena. The manual covers a complete
description of the experiments and exercises. The suggested experiments
cover almost all the units/ topics including those on diversity in living world,
plant, animal and human physiology, genetics, bio-technology and human
welfare and environment. A standard format has been used to describe each
experiment which includes
Aim: It gives a brief title of the experiment under investigation.
Principle: It is a very brief introduction of the experiment under
investigation and explains the biological phenomenon involved. It gives
brief but comprehensive ideas about the design of the experiment and
explains the significance of the phenomenon being studied.
Materials required: This includes the names of plants/ animals to be
used as 'samples', the type of apparatus, the type and quantity of
glasswares required, reagents, chemicals and solutions needed, their
concentration and other specifications, method of preparations of
solutions and reagents. If a particular material/ chemical/ glassware is
not available, sufficient alternatives have been suggested.
Procedure: This section includes full details of experimental procedure
explained stepwise, including special precautions necessary to be taken
while the experiment is being conducted. Drawings of the samples,
apparatus and the experimental setup, wherever found necessary, have
been included to facilitate the students to perform the experiment as
accurately as possible.
INTRODUCTION


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4
LABORATORY MANUAL: BIOLOGY
Observation and Results: This section deals with the recording of all
observations made during the experiment. Students are advised to
consider the entire data. Data can be represented in the form of tables,
graphs and histograms wherever possible. Use of units in which various
quantities are measured has been indicated in the manual.
Discussion: Included in this heading is a statement of the conclusions
drawn from the experimental results and compared thesis (wherever
possible) with any comparable data from other sources. The relevance
of the conclusions drawn from the experimental results to the various
processes under investigation and to the life of plant, animal and microbes
has been prompted out.
Precaution: This section contains all the necessary precautions to be
taken during experimentation to obtain results free of errors. However,
attempts have been made to mention required precautions along with
the procedure also.
A great emphasis has been laid on a student getting valid results and
interpreting them. It is essential that the teacher should properly explain
each experiment so that inexperienced students will be able to obtain accurate
results within a reasonable time. Teachers are also expected to help students
in identifying errors and mistakes committed during experiments and ways
for correcting them. It is possible that some of the students may undoubtedly
be capable of doing more sophisticated work than that represented in the
manual. But introductory course of this sort has been designed to help all
students for some useful and joyful experience by conducting the
experiments of their own. The manual also aims that students and teachers
not be discouraged by either incomplete experiments or experiments which
yield apparently meaningless results.
With the objectives of inculcating scientific temper among learners and
providing them an opportunity to undertake independent scientific
investigation, Investigatory Project Work has been included as an integral
part of the practical curriculum of Class XII. Such investigatory projects
are expected to provide thrill in the learning process. It is also expected to
serve the real purpose of practicals, i.e., developing an ability to hypothesise
and design experiments to address certain problems, to make observations
methodically and to draw conclusions out of the experimental data. A
comprehensive idea about undertaking investigatory project has been given
in the book with a list of a few problems on which investigatory project
work can be undertaken. However, the list is only suggestive and considering
the wider scope students can undertake any kind of investigatory project
work depending on their region, its climatic condition, availability of
resources, etc.


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Aim: To study the reproductive parts of commonly available flowers
Principle: The male reproductive parts of a flower are the stamens collectively called
androecium and the female reproductive parts are the carpels/ pistils collectively called
gynoecium. The individual units of stamen consist of a filament, which supports the anther
lobes. Gynoecium consists of stigma, style and ovary. Many variations are found in different
characteristics of both the stamens and carpels. We shall try to study these variations in the
reproductive parts of flowers in the exercise.
Requirement: Commonly available flowers, needles, forceps, razor/ scalpel blade, brush, slides,
cover slip, watch glass, magnifying lens, dissecting microscope, compound microscope, etc.
Procedure
(i) Familiarise with the terms to describe the reproductive parts of flowers
given in annexures of Exercise No. 11 of Laboratory Manual: Biology
(Class XI) and at the end of this experiment.
(ii) Observe the flower with the naked eye, hand lens or under a dissecting
microscope. Study their reproductive parts and count the number of
stamens and record their cohesive and adhesive features.
(iii) Cut L.S. of the flower and place it on a slide to observe the following
characters:
(a) Placement of anthers
(b) Position of the ovary: epigynous/ perigynous/ hypogynous.
(iv) Mount one stamen on a slide and study the following characters:
(a) Attachment of filament to anther
(b) Dehiscence pattern of the anther lobes for discharge of pollen.
(v) Cut T.S. of anther lobe to observe the number of pollen sacs.
(vi) Mount the pistil on a slide and study style, stigma and ovary. Record
the number of stigma and nature of pistil.
(vii) Cut T.S. of ovary, mount it on a slide and observe
(a) Number of locules in the ovary
Exercise 1


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6
LABORATORY MANUAL: BIOLOGY
(b) Type of placentation
(c) Number of ovules per locule
(viii) Draw labelled figures of your preparation and observations.
Questions
1. Name the most common type of placentation observed.
2. What is the most common type of dehisence pattern in anthers?
3. Name a few unisexual flower-bearing plants studied by you.
4. Flower is a modified shoot. J ustify the statement based on your observation.


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EXERCISE 1
Annexure 1
Description of reproductive parts of flowers
Androecium
Number of stamens The number of stamens may vary from a few to many in dif-
ferent flowers
Stamens may be free or united. I f united they can be of the
following type:
(i ) Syngenesi ous: Filaments free and anthers united,
e.g., Sunflower.
(i i ) Synandr ous: Stamens fused all through their length,
e.g., Cucurbita
(i i i ) Adel phous: Anthers remain free and filaments are united.
Adelphous condition can be
(a) MonoadelphousUnited to form 1 bundle, e.g.,
China rose
(b) DiadelphousUnited to form 2 bundles, e.g., Pea
(c) PolyadelphousUnited into more than two
bundles, e.g., Lemon
Fusion of stamens with other parts of the flower
(i ) Epi petal ous: Stamens fused with petals,
e.g., Sunflower, Datura
(i i ) Epi phyl l ous: Stamens fused with perianth,
e.g., Lily
(i ) Basi f i xed: Filament attached to the base of anther,
e.g., Mustard
(i i ) Adnate: Filament attached along the whole length of
anther, e.g., Michelia, Magnolia
(i i i ) Dor si f i xed: Filament attached to the back of anther,
e.g., Passion flower
(i v) Ver sati l e: Anther lobes attached with filament in the
middle portion with both ends free, e.g., Gramineae
family
(i ) Por ous: Pollens released through pores, e.g., Brinjal,
Potato
(i i ) Longi tudi nal : Pollens released through the longitudinal
slit of another lobes, e.g., China rose, Cotton
Cohesion
(Fig. 1.1 ae)
Adhesion
(Fig. 1.2)
Attachment of filament to
anther
(Fig.1.3 ad)
Dehiscence pattern
(Fig. 1.4 a,b)


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LABORATORY MANUAL: BIOLOGY
Gynoecium
Number of stamens The number of stamens may vary from a few to many in
different flowers
(i) Epi gynous: Position of ovary inferior to other floral parts,
e.g., Mustard, China rose
(i i ) Per i gynous: Other floral parts are attached around the
ovary, e.g., Apple, Guava
(i i i ) Hypogynous: Position of ovary superior to other floral
parts, e.g., Sunflower
I f number of carpels is more than one, they may be
(i ) Apocar pous: Carpels are free. Each carpel has its own
style and stigma, e.g., Rose
(i i ) Syncarpous: Carpels are united, e.g., Ladys finger, Tomato
Vary from one to many
(i ) Uni l ocul ar : One locule, e.g., Rose, Pea
(i i ) Bi l ocul ar: Two locules, e.g., Datura
(i i i ) Mul ti l ocul ar : Many locules, e.g., Ladys finger, China rose
(i ) Mar gi nal : The placenta forms a ridge along the ventral
suture of the ovary and the ovules are borne on this ridge,
e.g., Pea
(i i ) Axi l e: The ovary is partitioned into several chambers or
locules and the placentae are borne along the septa of
the ovary, e.g., Tomato, China rose
(i i i ) Par i etal : The ovules develop on the inner wall of the
ovary or on peripheral part. Ovary unilocular but in some
cases becomes two chambered due to formation of a false
septum, e.g., Mustard
(i v) Fr ee centr al : Ovules are borne on the central axis and
septa are absent, e.g., Carnation, Chilly
(v) Basal : Placenta develops at the base of the ovary,
e.g. ,Sunflower.
Position of ovary
(Fig. 1.5 ad)
Cohesion
(Fig. 1.6 ac)
Number of locules in ovary
Placentation
(Fig. 1.7 ae)


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EXERCISE 1
Fig.1.1 Cohesion of stamens: (a) Syngenesious (b) Synandrous
(c) Monoadelphous (d) diadelphous (e) Polyadelphous
(a) (b) (c)
(d)
(e)
Fig.1.2 Adhesion of Stamens: Epipetalous/ Epiphyllous
Fig.1.3 Attachment of fi l ament to anther: (a) Basi fi xed (b) Adnate
(c) Dorsifixed (d) Versatile
(a) (b) (c) (d)


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LABORATORY MANUAL: BIOLOGY
Fig.1.5 Position of ovary: (a) Epigynous (bc) Perigynous (d) Hypogynous
(b) (c) (d)
Fig.1.4 Dehiscence pattern of anther: (a) Porous (b) Longitudinal
(a) (b)
Fig.1.6 Cohesion of carpels: (a) Apocarpous (bc) Syncarpous
(a) (b) (c)
(a)


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EXERCISE 1
Fig.1.7 Placentation: (a) Marginal (b) Axile (c) Parietal (d) Free central (e) Basal
(a) (b) (c)
(d) (e)


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Aim: To calculate percentage of pollen germination
Principle: In nature, pollen grains germinate on the compatible stigmas of the carpel. Pollen
grains can also be induced to germinate in a synthetic medium. During germination, intine
(inner wall) of pollen grain emerges out as pollen tube through one of the germ pores in exine
(outer wall).
Requirement: Calcium nitrate, boric acid, sucrose, distilled water, petridish, slides, coverslips,
brush, needle, microscope, and mature pollen grains of Tradescantia/ balsam/ Jasmine/ lily/
pomegranate/ grass/ Vinca/ China rose/ Petunia.
Exercise 2
Procedure
(i) Prepare the pollen germination medium by dissolving 10g sucrose,
30mg calcium nitrate and 10mg boric acid in 100ml of distilled water.
Alternatively 10% sucrose solution can also be used.
(ii) Take a drop of medium or 10% sucrose solution on a cover slip and
sprinkle mature pollen grains on the drop.
(iii) Invert the cover glass on to a slide
(iv) After 10 minutes, observe the slide under microscope.
(v) Count (a) total number of pollen grains seen in the microscope field,
and (b) the number of pollen grains that have germinated.
Observation
Several pollen grains germinate and put forth pollen tubes. Count the total
number of pollen grains and the number of germinated pollen grains in 3-5
different microscope fields. Tabulate your observations and calculate the
percentage of pollen germination.
Name of the plant used as source of pollen
Number of pollen grains in a field of microscope =N
Number of germinated pollen grains in a field of microscope =n
Percent pollen germination =
100
n
N
or
100
N
n


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EXERCISE 2
Discussion
Although pollen grains of many species germinate in this medium, the
percentage of germinations and the time taken for germination varies in
different species. Draw a germinating pollen grain and label.
Number of Total number of pollen (N) Total number of pollen germinated (n) % pollen germination
observation
100
n
N
1.
2.
3.
4.
5.
Average
Questions
1. How many pollen tubes emerge from a pollen grain?
2. What does the pollen tube carry?
3. Can you explain as to why some pollen grains fail to germinate?
4. Why do we use sucrose as the medium for pollen germination?


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Aim: To study pollen tube growth on stigma
Principle: Pollen grains germinate and form pollen tubes after they get deposited by the process
of pollination on compatible stigma. Pollen tube, made up of cellulose, is an extension of the
inner wall of pollen grain (intine). It emerges through one of the germ pore and passes through
tissues of stigma and style to reach the ovule. The growing pollen tube is observed by staining
with cotton blue.
Requirement: 56 excised styles with stigma of Petunia/ grass/ maize/ sunflower/ Abelmoschus
(Lady's finger), beaker, water, slides, cover slips, cotton blue stain, microscope, brush, needle.
Exercise 3
Fig.3.1 Growth of pollen tube
in the style of a carpel
Pollen grains
Pollen tube
Styl e
Procedure
(i) Place the stigmas in boiling water in a beaker
for softening the tissues for 510 minutes.
(ii) Stain with cotton blue for 35 minutes and
wash with water to remove excess stain.
(iii) Mount one stigma in a drop of glycerine on a
slide. Place a cover slip on the stigma and gently
press the cover slip on the material. Observe
the slide under a microscope.
(iv) If you fail to observe pollen tubes mount
another stigma.
Observation
Look for long blue-coloured tubular structures
traversing through the tissues of stigma and
style (Fig. 3.1).


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EXERCISE 3
Discussion
Pollen tubes are seen amidst the stylar tissue. Many pollen tubes may be
seen. Trace the origin of pollen tubes to the pollen grains present around
the surface of the stigma.
Questions
1. Can pollen grains of one plant species germinate on stigma of other species?
Give reasons.
2. Do all pollen tubes reach the ovules?
3. Are all the pollen tubes of equal length? If not, why?


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Aim: To study the discrete stages of gametogenesis in mammalian testis and ovary
Principle: In all male and female organisms gamete formation takes place in their gonads, i.e.,
testis and ovary respectively. The process of gamete formation, called gametogenesis involves
meiotic cell division. The gametogenic development in testis is called spermatogenesis and in
ovary it is oogenesis. They exhibit marked differences and can be examined in transverse section
(T.S.) of these organs.
Requirement: Permanent slides of T.S. of testis and ovary, compound microscope,
lens-cleaning paper and cleaning fluid
Procedure
(i) Clean the slide and microscopes eye and objective lenses with the
help of lens cleaning paper using any cleaning fluid.
(ii) Place the slide on the stage of the microscope and observe first under
lower magnification and then in higher magnification. Observe various
stages of gamete development.
(iii) Record your observations in the notebook and draw labelled diagrams.
Observation
T.S. of testis
Exercise 4
Fig. 4.1 T.S. of mammalian testis
Seminiferous tubule
Spermatozoa
Germinal
Epi thel i um
Spermatogonia
(i) You will observe a large number
of seminiferous tubules under
lower magnification. Observe a
compl ete tubul e i n hi gher
magnification and view various
stages of gamete development
from periphery towards lumen
(Fi g. 4.1) and i denti fy the
following types of cells namely,
Germi nal epi thel i um,
Spermatogonial cells, Primary
spermatocytes, Secondary
spermatocytes, Spermatids and
Spermatozoa.


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17
EXERCISE 4
(ii) In T.S. of testis the space between tubules are filled with blood vessels
and a specific cell type called Leydig's cell or Interstitial cells.
T.S. of Ovary
(i) In the section of ovary, there is a mass
of ti ssue l i ned wi th germi nal
epithelium. I nside that you will
observe an ovum, which is a cell
surrounded by one to several layers
of fol l i cul ar cel l s. As the ovum
matures, the number of surrounding
follicular cell layer increases (Fig. 4.2).
(ii) I n the l ater stage of fol l i cul ar
development a cavity called antrum
appears.
(iii) The cavity gets further enlarged and
the follicle grows bigger. This is the
stage of Graafian follicle ready to release the ovum (ovulation).
(iv) In the next stage, you may notice a Corpus luteum, and/ or Corpus
albicans, which differ from each other and also from Graafian follicle in
their features.
Discussion
Spermatogenesis is a continuous process after attainment of puberty, and
that is why gamete development and spermatozoa are observed in a single
seminiferous tubule. In case of ovary, the follicular development stages are
observed.
Questions
1. What would happen if meiosis fails to occur in gametocyte?
2. At which stage of follicular development, is ovum released?
3. Spermatogenesis is a continuous process. J ustify the statement.
4. Draw a labelled diagram of T.S. of testis.
5. Draw a labelled diagram of T.S. of ovary.
6. What would happen if sperms are devoid of their tail?
7. What are the consequences of failure of ovulation?
Fig. 4.2 Section of mammalian ovary
Graafian Follicle
Corpus luteum
Corpus albicans
Antrum


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Aim: To study and identify various stages of female gametophyte development in the ovary
of a flower
Principle: In flowering plants, female gametophyte (embryo sac) is a microscopic structure
situated deep inside the ovule. An ovule generally has one female gametophyte. Development
of female gametophyte begins with megaspore mother cell. Most common type of female
gametophyte is the monosporic, 8-nucleate, 7-celled type.
Requirements: Permanent slides of V.S. of ovary, photographs/ chart or models showing
stages of female gametophyte development and microscope
Procedure
(i) In a V.S. of ovary we generally find several ovules. Carefully observe
each ovule and locate as many stages of female gametophyte
development as possible.
(ii) Draw the diagrams as observed under microscope.
Exercise 5
Embryo sac
Fig. 5.1 V.S. of an ovule
Chal aza
Outer integument
Inner integument
Micropyle
Funiculus
Observation
(i) Record the features of ovule
like number of integuments,
nucellus and micropylar
and chalazal poles. Fig 5.1
shows the femal e
gametophyte (embryo sac)
as seen in a V.S. of an ovule.
Di fferent stages of
devel opment of femal e
gametophyte are shown in
Fig. 5.2.
(ii) Observe the placement of
embryo sac close to the
micropylar pole.


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EXERCISE 5
Fig. 5.2 Stages of gametophyte development: (a) megaspore with 2 nucleus
(b) 4- nucl eate stage (c) 8- nucl eate stage (d) 8- nucl eate stage
showing 3+2+3 distribution of nuclei (e) mature embryo sac.
(a) (b)
(c)
Egg
Synergids
Central cell
Secondary nucleus
Anti podal s
(d) (e)
(iii) Note the contents of embryo sac, namely, an egg apparatus
(2 synergids and egg) at micropylar end, secondary nucleus in the
center and three antipodal cells at the chalazal end (Fig. 5.2).
Questions
1. Explain the difference between gamete and a gametophyte.
2. Name two differences between synergids and egg.
3. What is the function of polar nuclei?


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Aim: Preparation and study of mitosis in onion root tips
Principle: Somatic growth in plants and animals takes place by the increase in the number of
cells. A cell divides mitotically to form two daughter cells wherein the number of chromosomes
remains the same (i.e., unchanged) as in the mother cell. In plants, such divisions rapidly take
place in meristematic tissues of root and shoot apices, where the stages of mitosis can be
easily observed. In animals, mitotically dividing cells can be easily viewed in the bone marrow
tissue of a vertebrate, epithelial cells from gills in fishes and the tail of growing tadpole larvae
of frog.
Requirement: Onion bulbs, wide mouth glass tubes/ jar/ bottle, glacial acetic acid, ethanol
2-4% acetocarmine/ acetoorcein stain, N/ 10 HCl, spirit lamp/ hot plate, slide, cover slips,
blotting paper, molten wax/ nail polish and compound microscope
Procedure
Growing of root tips
Select a few medium-sized onion bulbs. Carefully remove the dry roots
present. Grow root tips by placing the bulbs on glass tubes (of about 34
cm. diameter) filled with water. Care should be taken so that the stem portion
of the bulb (basal part) just touches the water. A few drops of water may be
added periodically to compensate evaporation losses. New roots may take
36 days to grow. Cut 23 cm long freshly grown roots and transfer them to
freshly prepared fixative, i.e., aceto-alcohol (1:3:: glacial acetic acid : ethanol).
Keep the root tips in the fixative for 24 hours and then transfer them to 70%
ethanol (for preservation and use in future). Onion root-tip cells have a cell
cycle of approximately 24-hour duration, i.e., they divide once in 24 hours,
and this division usually takes place about two hours after sunrise. Therefore,
roots grown on water should be cut only at that time to score maximum
number of dividing cells.
Preparation of slide
Take one or two preserved roots, wash them in water on a clean and grease-
free slide. Place one drop of N/ 10 HCl on the root tip followed by 23 drops
of aceto-carmine or aceto-orcein stain on it. Leave the slide for 510 minutes
Exercise 6


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21
EXERCISE 6
on a hot plate (or warm it slightly on spirit lamp). Care should be taken that
the stain is not dried up. Carefully blot the excess stain using blotting paper.
Now cut the comparatively more stained (23 mm) tip portion of the root
and retain it on the slide and discard the remaining portion. After
(1020 seconds) put one or two drops of water and blot them carefully using
blotting paper. Again put a drop of water on the root tip and mount a cover
slip on it avoiding air bubbles. Place the slide in between the folds of blotting
paper using the fingers in such a way that the cover slip mounted on the
slide is properly held. Now slowly tap the cover slip using the blunt end of a
pencil so that the meristematic tissue of the root tip below the cover slip is
properly squashed and spread as a thin layer of cells. Carefully seal the
margins of the cover slip using molten paraffin wax or nail polish. This
preparation of onion root tips cells is now ready for the study of mitosis.
Study of slide
Place the slide on the stage of a good quality compound microscope. First
observe it under the lower magnification (10 X objective) to search for the
area having a few dividing cells. Examine the dividing cells under higher
magnification of the microscope to observe the detailed features of mitosis.
Observation
The stages of mitosis can be broadly categorised into two parts: karyokinesis
(division of nucleus) followed by cytokinesis (division of cytoplasm, and
ultimately of the cell). Those cells, which are not in the phases of cell division
are considered to be in interphase. You may observe that most of the cells
in a microscope field are in interphase
Interphase
The cells are mostly rectangular, oval or even circular in shape, with almost
centrally situated densely stained nucleus. The chromatic (coloured) material
of the nucleus is homogeneous and looks granular. The boundary of the
nucleus is distinct. One or few nucleoli (sing: nucleolus) can also be observed
inside the nucleus (Fig. 6.1a).
Stages of Mitosis
(a) Prophase
Intact nuclear outline is seen. The chromatin (seen as a homogeneous
material in the nucleus at interphase) appears as a network of fine threads
(chromosomes). Nucleoli may or may not be visible (Fig. 6.1b).


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LABORATORY MANUAL: BIOLOGY
If the cell under observation is in the early stage of prophase then the
chromatin fibres (chromosomes) are very thin. However, in the cells
at late prophase, comparatively thicker chromatin fibres would be
visible. Besides this, in the late prophase the nuclear membrane
may not be noticed.
(b) Metaphase
The nuclear membrane disappears. Chromosomes are thick and are seen
arranged at the equatorial plane of the cell (Fig. 6.1c). Each chromosome at
Fig.6.1 I nterphase (a) and stages of mitosis (b - e) actual microscopic
vi ew on l eft si de and i ts di agrammati c representati on on the
right hand side
a. Interphase
b. Prophase
c. Metaphase
d. Anaphase
e. Telophase


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EXERCISE 6
this stage has two chromatids joined together at the centromere, which can
be seen by changing the resolution of the microscope. Nucleolus is not
observed during metaphase.
(c) Anaphase
This stage shows the separation of the chromatids of each chromosome. The
chromatids separate due to the splitting of the centromere. Each chromatid
now represents a separate chromosome as it has its own centromere. The
chromosomes are found as if they have moved towards the two poles of the
cell. The chromosomes at this stage may look like the shape of alphabets 'V',
'J ' or 'I' depending upon the position of centromere in them. Different anaphase
cells show different stages of movement of chromosomes to opposite poles,
and they are designated to represent early, mid and late anaphase (Fig. 6.1d).
(d) Telophase
Chromosomes reach the opposite poles, lose their individuality, and look
like a mass of chromatin (Fig. 6.1e). Nuclear membrane appears to form the
nuclei of the two future daughter cells.
Cytokinesis
In plants, a cell plate is formed in the middle after telophase. The plate can
be seen to extend outwards to ultimately reach the margin of the cell and
divide the cell into two. Such cell plates are characteristic of plant cells
(Fig. 6.2). However, in an animal cell, the two sides of the cell show inpushings
or constrictions formed from the peripheral region in the middle of the cell,
which grow inward and meet to divide the cell into two daughter cells.
Draw labelled diagrams of all the phases of mitosis.
Fig. 6.2 Cytokinesis
Discussion
Mitotic index (MI) is defined as a ratio of the total number of dividing cells (n)
and the total number of cells (N) in a particular focus chosen randomly under
the microscope and is calculated as MI =
100
n
N
. By randomly selecting
5 to 10 such foci, one can estimate the mitotic index for a given type.


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LABORATORY MANUAL: BIOLOGY
Questions
1. Suggest names of a few tissues, which are suitable for the study of mitosis.
2. Why is mitosis also known as equational division?
3. What shape would a metacentric and a sub-metacentric chromosome exhibit
during the anaphase stage?
4. How does cytokynesis differ in plant and animal cells?
Features Interphase Karyokinesis Cytokinesis
Prophase Metaphase Anaphase Telophase
1. Cell morphology
2. Nuclear morphology
3. Chromosomes/ chromatids
Tabulate your observations in the tabular form given below
The effect of different samples of water (polluted or contaminated) can be
assayed on the mitotic-index (an indicative feature of somatic growth rate in
them).
Further, the impact of different types of pollutants on different phases of
mitosis can also be assayed.


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Aim: Study of stages of meiosis using permanent slides
Principle: Meiosis is a type of cell division in which the number of chromosomes is halved
(from diploid to haploid) in the daughter cells, i.e., the gametes. The division is completed in
two phases, meiosis I and meiosis II. Meiosis I is a reductional division in which the chromosomes
of homologous pairs separate from each other. Meiosis II is equational division resulting in the
formation of four daughter cells. Stages of meiosis can be observed in a cytological preparation
of the cells of testis tubules or in the pollen mother cells of the anthers of flower buds.
Requirement: Permanent slides of meiosis and compound microscope
Exercise 7
Procedure
Place the slide on the stage of the microscope and search for the dividing
cells using lower magnification. When dividing cells are located observe them
under higher magnification.
Observation
Observe various stages of meiosis and identify them on the basis of the specific
features given in the table 7.1. A significant number of cells will be in the
Interphase. These cells have a centrally positioned densely stained nucleus.
In case of slide of animal tissue a few mitotically dividing spermatogonial
cells may also be seen.
Unlike the prophase of mitosis, it is a comparatively complex phase
characterised by a number of events. Five sub-phases can be
identified in it.
(a) Leptotene (leptos =slender tene =band or thread)
(i) The nuclear membrane and nucleolus are not distinctly
observable (Fig. 7.1 a).
(ii) Fine network of thin threads are seen uniformly distributed
in the nucleus.These are chromatin threads, which may be
observed as more prominent structures in the later stages.
Meiosis I
1. Prophase I
Table 7.1 Different stages of meiosis and their features


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LABORATORY MANUAL: BIOLOGY
Fig. 7.1 Sub-phase of Prophase I (a-d) actual mi croscopi c vi ew on l eft
side and its diagrammatic representation on the right hand side
(a) Leptotene
(b) Zygotene
(c) Pachytene
(d) Diplotene-Diakinesis


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EXERCISE 7
2. Metaphase I
3. Anaphase I
4. Telophase I
Meiosis II
1. Prophase II
(b) Zygotene (Zygon = paired)
This stage is characterised by the pairing of the homologous
chromosomes, which can be seen as paired chromatin threads
(bivalents) (Fig. 7.1b).
(c) Pachytene (pachy = thick)
The chromatin threads get condensed and appear shortened and
thick. Pairs of homologous chromosomes can be seen. Each
chromosome has two chromatids and thus each bivalent consists
of four chromatids. This configuration is called tetrad (Fig. 7.1c).
(d) Diplotene (diplos =double)
The homologous chromosomes (each made up of two chromatids)
show distinct separation from each other except at few regions where
attachments are seen (Fig. 7.1d). These are chiasmata (sing.
chiasma) representing the site of exchange of the parts between
two homologous chromosomes (i.e. crossing over).
(e) Diakinesis (Dia =opposite; kinesis=separation or movement)
(i) The homologous pair of chromosomes appear more shortened,
thick and prominent (Fig. 7.1d).
(ii) Chiasmata can be still observed.
(iii) All the homologous pairs appear scattered in the cell.
Homologous chromosomes are still in pairs, and are arranged along
the equatorial plane of the cell (Fig. 7.2a). At this stage, the number
of bivalents can be counted. Chiasmata may still be seen in a few
bivalents.
The chromosome pairs appear to have moved towards the two
opposite poles of the cell. At the later stage, the anaphase - I may
show the assembly of chromosomes at two poles (Fig. 7.2b). This
results into the reduction of number of chromosomes to half.
This stage can be identified by the presence of two chromatids in
each chromosome.
The chromosomes present at the two poles appear decondensed
and form two distinct nuclei (Fig. 7.2c).
Note: After the telophase I stage there may or may not be cytokinesis.
Thereafter the cell enters into the second meiotic division.
(i) Distinct thread- like chromatin fibres or rod- shaped chromosome
are seen.


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LABORATORY MANUAL: BIOLOGY
Fig.7.3 Phases of Meosis I I (a,b) actual microscopic view on left side
and its diagrammatic representation on the right hand side.
(a) Metaphase I I
(b) Anaphase I I
(a) Metaphase I
(b) Anaphase I
(c) Telophase I
Fig. 7.2 Phases of Meosis I (a-c) actual microscopic view on left side
and its diagrammatic representation on the right hand side.


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EXERCISE 7
2. Metaphase II
3. Anaphase II
4. Telophase II
Questions
1. What is the significance of meiosis?
2. What is synapsis and crossing over?
3. How can anaphase I and anaphase II be distinguished from each other?
4. Indicate distinguishing feature of metaphase I of meiosis and metaphase of mitosis.
5. How many daughter cells are produced at the end of meiosis?
6. The daughter cells produced at the end of meiosis are genetically different. Explain.
7. What is the significance of synapsis?
This phase is similar to that of mitotic division
(i) The chromosomes having two chromatids attached at the
centromere are observed arranged at the equatorial plane of the
cell.
Note: Metaphase II of meiosis can be differentiated from metaphase-I
on the basis of the following features:
(ii) Each chromosome of metaphase I I has two chromatid
(Fi g. 7.3a) whereas i n metaphase I these are pai red
homologous chromosomes each having two chromatids thus
forming tetrad.
(iii) In the metaphase I of meiosis, a few chiasmata are observed,
where as no chiasmata are observed during metaphase II.
The two chromatids of each chromosome after separation appear
to lie at the two poles of the cell (Fig. 7.3b).
Note: Anaphase II can also be distinguished from the anaphase I of
meiotic division on the basis of chromatids: In anaphase I, each
chromosome has two distinct chromatids, but in anaphase II, each
chromosome is represented by one chromatid only.
The separated chromosomes appear decondensed and form nuclei
(Fig. 7.3c).


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Aim: To study the blastula stage of embryonic development in mammals, with the help of
permanent slide, chart, model or photograph
Principle: The zygote undergoes a few cycles of mitotic divisions to form a solid ball of cells called
morula. The cells continue to divide and at a later stage a cavity is formed within it. This stage is
blastula. The internal structural details of blastula can be observed in its transverse section.
Requirement: Permanent slide, chart/ model of T.S. of blastula, compound microscope,
lens cleaning fluid and paper
Exercise 8
Procedure
Observe the slide under lower magnification of the microscope. In case of
chart/models/photographs, note the feature of blastula in your practical
record and draw labelled diagram.
Observation
In transverse section, the blastula appears as a sphere with a cavity, called
blastocoel within it (Fig. 8.1). Notice an outer layer of blastomeres called
trophoblasts. A cellular mass, adhered to the trophoblast is present on one
end of the blastula. It is called inner cell mass.
Fig.8.1 Blastula stage of a mammal


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31
EXERCISE 8
Questions
1. What are the differences between blastula and morula?
2. What are the main structures you observe in T.S. blastula?
3. Match the stages in column I with features in column II
Column I Column II
(a) Trophoblast (i) Dividing cells of the morula
(b) Morula (ii) Outer layer of blastula
(c) Blastocoel (iii) Solid ball of cells
(iv) Cavity


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Aim: To verify Mendel's Law of Segregation
Principle: When two pure lines with contrasting forms of a particular character (phenotypes) are
crossed to produce the next generation (F
1
generation), all the members of the progeny are of only
one phenotype i.e. of one of the two parents. The phenotype that appears is called dominant, and
the one that does not appear is called recessive. When the F
1
plants are selfed, the progeny i.e. the
F
2
generation is in the ratio of 3 dominant: 1 recessive (: or 75%: 25%). This reappearance of
the recessive phenotype in F
2
generation verifies law of segregation.
Requirement: 64 yellow and 64 green plastic beads, all of exactly same shape and size, (when
beads are not available, pea seeds may be coloured using paint, these beads represent the
gametes of a specific trait), plastic beakers/ petri dishes and a napkin/ hand towel
Procedure
Students have to work in pairs to perform the experiment. The following
steps are to be strictly followed in the sequence mentioned below.
(i) Put 64 yellow beads in one beaker/petridish and 64 green beads in
the other to represent respectively male and female gametes. Let the
yellow bead be indicated by Y and green bead by y.
(ii) Take a bead from each container and place them together (it represents
fertilisation) on the napkin spread before you on the table. (One student
to take out beads and to put in the hands of the other student who will
put them on the table).
(iii) Just like the previous step, continue to pick beads and arrange them
in pairs. Thus 64 pairs of beads are obtained representing the 64
heterozygous F
1
progeny.
Note that all the F
1
individuals are represented by one yellow and one
green bead.
(iv) Put 32 F
1
progeny in one petridish and the remaining 32 in another
petridish (representing the F
1
males and females).
(v) Stir the beads of each petridish with a pencil/pen for about 10 times
taking care that no bead falls off.
Exercise 9


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33
EXERCISE 9
(vi) To obtain the F
2
generation, one student would withdraw one bead
from one beaker labelled male and one from the other beaker labelled
female keeping his/her eyes closed (to ensure randomness), and put
them together in the stretched palm of the partner, who will put them
together on the napkin spread over the table. Continue this process till
all the beads are paired. Thus 64 offsprings of F
2
are obtained.
(vii) Note the genotype (YY or Yy or yy) of each pair, and their possible
phenotype.
(viii) Have six repeats of the experiment (steps i to vii) with partners changing
their roles. Pool all the data from the six repeats together.
(ix) Calculate the genotypic and phenotypic ratios of your pooled data.
Note that larger the sample size, more accurate is the result.
Observation
Record the result in the following table:
Generation Repeat No. Total no. of Genotype (s) Phenotype (s)
individuals YY Yy yy
F
1
1.
2.
3.
4.
5.
6.
Total
F
2
1.
2.
3.
4.
5.
6.
Total
Phenotypic Ratio: in F
1
.
in F
2
.
Genotypic Ratio: in F
1
.
in F
2
.


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LABORATORY MANUAL: BIOLOGY
Questions
1. Do you expect the same results in terms of 3:1 ratio in F
2
if you had started with
smaller number of beads (say 10 beads)?
Discussion
The results are so because each diploid individual contains two copies of
every gene - one copy on each of the two homologous chromosomes. These
two copies of the gene may be of similar type (YY or yy) or are dissimilar Yy.
The former (YY or yy) are called homozygous for that particular character,
and the Yy are called heterozygous ones. The pure lines in the above cross
are homozygous ones, which contributed only one copy of their gene (as a
result of meiosis) to their F
1
progeny to restore its diploid nature with genotype
Yy (heterozygous) where only one form (allele) is expressed (dominant) and
the other form (allele) is not expressed (recessive). This is the phenomenon
of Dominance.
When the F
1
individuals are crossed together to raise the F
2
generation,
each F
1
individual produces two types of gametes: 50% having dominant
allele, and the remaining 50% having recessive allele. These gametes undergo
random fusion during fertilisation to produce the F
2
generation. According
to simple probability of mixing of opposite sex gametes (sperms and ova),
offsprings of three genotypes are likely to appear as follows: [(half of gametes
of Y type + half of remaining gamete y type) X (half gametes of Y type + half
of remaining gamete of y type)] = One-fourth of F
2
individuals of YY phenotype
+ half of F
2
individual Yy type + one-fourth of F
2
individul of yy type. Among
these proportion of dominant phenotype would be YY+ Yy = yellow
and recessive phenotype yy i.e. green phenotypes in 3:1 or 75%:25%
ratio.
This ratio of 3:1 in the F
2
suggests that in the F
1
heterozygotes, the
recessive allele does not get destroyed and remains only in the recessive
(dormant) state to get an opportunity to express itself when it has separated
from the influence of the dominant allele (Y). This is called Law of Segregation
of the alleles.

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Aim: To verify the Mendels Law of Independent Assortment
Principle: In a dihybrid cross, the segregation of one gene pair is independent of the segregation
of the other pair. It means that genes of two different traits assort independently to give a
probability ratio equal to segregration probability ratio of one allele pair X segregation probability
ratio of other allele pair, which comes to, (3:1) X (3:1) = 9:3:3:1
Requirement: Plastic beakers; 64 plastic beads each of yellow, green, red and white to represent,
yellow and green colour of seed coat and red and white flowers respectively and napkin/ hand
towel
Exercise 10
Procedure
Students are to work in pair.
The following steps are to be followed sequentially:
(i) Place 64 beads of each colour in four separate beakers.
(ii) Put the beakers containing the yellow and red beads on your left side,
and those containing the green and white beads on your right side.
The beakers on your left side represent plants bearing yellow seed and
red flower (dominant character YY, RR). Beakers on the right side
represent plants bearing green seeds and white flowers (recessive
character yy, rr). These are the two parental types having contrasting
forms of two different characters.
(iii) Stir the beads in each beaker with a pencil/ pen. Each bead now
represents alleles in the male and female gametes.
(iv) Pick up one yellow, one green, one red and one white bead, and put
them together on the napkin spread on the table.
(v) Continue picking up and putting together of the beads of all colours
as mentioned in the previous step, till all the beads are utilised.
(vi) Note that in all, 64 such 4-bead clusters are obtained representing the
F
1
individuals. Ascertain their genotype and phenotype.
(vii) Next step is to cross these F
1
individuals to raise the F
2
generation. Let
us suppose half of the 4-bead clusters (32 clusters) represent the male
parents and the remaining half (32 clusters) the female parents. Now
put the 32 red and 32 white beads together in one beaker (numbered-
I), and similarly put 32 yellow and 32 green beads together in other
36
LABORATORY MANUAL: BIOLOGY
beaker (numbered-II). These two beakers represent F
1
female. Similarly
put remaining 32 red +32 white beads in beaker numbered-III, and
32 yellow and 32 green one in beaker numbered-IV to represent the F
1
male. The arrangement can be presented as below
(viii) Stir the beads in each beaker with a pencil. In order to raise the F
2
generation, pick up (with eyes closed) one bead from the beaker-I of
female and one bead from the beaker-III of the male, and put into the
palm of the partner student. Similarly, pick up one bead each from the
beaker-II of female and beaker IV of male to put in the palm of the
partner. This partner would now keep all the four beads together (to
represent the F
2
individual). Continue this process till all beads are
utilised. At the end, 64 F
2
individuals (each represented by a 4-bead
cluster) are obtained.
(ix) Determine the genotype and phenotype of each of the 64 F
2
individuals
and write down the number of individuals of different genotypes and
phenotypes in the tabular form (given below), remembering that Y
(yellow seed colour) is dominant over y (green seed) and R (red flower)
is dominant over r (white flower).
(x) Repeat the whole procedure (steps i to ix) six times, and tabulate your
results.
Observation
Tabulate the results as follow:
Symbol (-) indicates the presence of corresponding dominant or recessive
allele e.g. Y or y and R or r.
Summarise your results (adding together the data of all the six repeats)
F1 Generation
(a) Total number of individuals: _________________________
(b) Phenotype (s) _________________________
(c) Genotype (s) _________________________
Female F
1
Male F
1
32 red +32 white (Beaker I) 32 red+32 white (Beaker III)
32 yellow +32 green (Beaker II) 32 yellow +32 green (Beaker IV)
37
EXERCISE 10
Generation Total No. Genotype Phenotype
& repeat of offsprings Y-R- Y-rr yyR- yyrr Yellow Yellow Green Green
No. Red White Red white
F1
1.
2.
3.
4.
5.
6.
Total
F2
1.
2.
3.
4.
5.
6.
Total
F2 Generation
(a) Total number of individuals _________________________
(b) Phenotypes _________________________
(c) Number of individuals in each phenotypic class:
Number Phenotype
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
(d) Phenotypic ratio _____________________
(e) Genotypic ratio _____________________
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LABORATORY MANUAL: BIOLOGY
Questions
1. Linked traits fail to assort independently. Explain.
2. How is independent assortment of alleles important from the point of view of
variation?
(f) Number of individuals of each genotypic class:
Number Genotype
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
(g) Genotypic Ratio ______________________________________
Discussion
The four phenotypic classes in the F2 generation are in ratio of 9:3:3:1 as
expected from the Law of Independent Assortment. The genotypic ratio
would be (1:2:4:2): (2:1):(2:1):1.
Note
1. In case six repeats of the experimental procedure are not feasible due
to time limitations, either the number of repeats be slashed down to
three or the data from single repeat of six different pair of students may
be pooled together to make the final calculations.
2. This Law of Independent Assortment was later found to be true only for
traits present on two different homologous pair of chromosomes, that
is, the two are not linked together. The linked traits do not assort
independently, rather they are inherited together (linked) except when
crossingover separates them.
3. It is quiet likely that you may not find your data exactly in the
expected ratio, instead almost approximate to it. The statistical
significance of this deviation from the exact expected ratio due to
probality can be checked using chi-square (
2
) test, about which
you will study in higher classes.
Aim: Preparation and analysis of Pedigree Charts
Principle: The Mendelian concept of dominance and segregation can also be studied in humans
by preparing and then analysing the pedigree charts. The internationally approved symbols for
indicating males and females, marriages, various generations (I, II, III), etc., are given below.
Exercise 11
Requirement: Information about characters/ traits in a family for more than one generation
Procedure
Select a family in which any one of the monogenic traits such as tongue
rolling, widow's peak, blood groups, red-green colour blindness, dimple in
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LABORATORY MANUAL: BIOLOGY
the cheek, hypertrichosis of ear, hitch-hiker's thumb, etc., is found. Ask the
person exhibiting the trait to tell in which of his/ her parents, grand parents
(both maternal and paternal), their children and grand children the trait in
question is present. Among surviving individuals the trait may also be
examined. The information made available is the basis for the preparation of
pedigree chart using the appropriate symbols. A careful examination of the
pedigree chart would suggest whether the gene for the character is autosome-
linked dominant or recessive, X - chromosome linked dominant or recessive,
Y- chromosome linked or not.
Explanation
1. Autosome Linked Dominant traits: These are the traits whose
encoding gene is present on any one of the autosomes, and the wild-
type allele is recessive to its mutant allele, i.e., the mutant allele is
dominant.
The pedigree-chart can be of the undernoted pattern (Fig. 11.2), where
the female being interviewed is exhibiting the trait, and is indicated by
an arrow-mark in the chart.
The characteristic features of inheritance of such type of traits are:
(a) Transmission of traits occurs from parents of either sex.
(b) Males and females are equally affected.
(c) The pedigree is vertical, i.e., the trait is marked to be present in each of
the generations.
(d) Multiple generations are characteristically affected.
Brachydactyly, polydactyly, dimple in the cheek are some of the common
traits of this type.
41
EXERCISE 11
2. Autosomal Recessive trait: These are the traits whose mutant allele is
recessive to its wild type allele.
The pedigree chart can be more or less of the pattern given below (Fig.
11.3), where the lady (marked by the arrow) is showing the trait. The bar
in the example represents the presence of corresponding dominant or
recessive allele for the specific trait.
Suppose the given trait is albinism. Denote its dominant allele as A
that produces pigments, and the recessive allele as a that fails to synthesise
the pigment, melanin. The female (our subject in generation III) is therefore
of genotype aa. She must have received each of her a allele from both the
parents (generation-II), who are therefore themselves normal but are definitely
of genotype Aa, and are carriers of the trait. The allele a must also have been
present in her grand parents too, of course in heterozygous condition also
to make them carriers (generation-I)
Albinism in the subjects children (generation-IV) suggests her husband
too to be of genotype Aa, a carrier. Marriage of her albino daughter to an
albino man is bound to produce all her grand-children albino (gen-V).
The following are the salient features of the inheritance of such type of traits.
(a) Occur in equal proportions in multiple male and female siblings, whose
parents are normal but carriers;
(b) The siblings are homozygous for the defective allele, but their parents,
though some may appear normal, are obviously heterozygous, i.e.,
are merely carriers of the trait.
(c) Consanguinity (marriage between man and woman genetically related
to each other, such as cousins) occasionally results in the appearance
of such traits.
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LABORATORY MANUAL: BIOLOGY
3. X-Linked Dominant traits: These are the traits whose encoding gene
is present on the X- chromosome, and the mutant allele of which is
dominant over its wild-type allele.
Such traits are very rare, and are almost difficult to find in the
population. One example is oral-facial-digital syndrome (Duchene
Muscular Dystrophy), which results in absence of teeth, cleft (bifid) tongue
associated with mental retardation. The pedigree chart may appear as
follows (Fig. 11.4):
The possible genotypes of the above pedigree can be written as follows
(Fig 11.5):
Fig. 11.5 Genotypes of individuals shown in Fig. 11.4
43
EXERCISE 11
Here, the dominant mutant allele is denoted by D, and its recessive wild
type allele is denoted by d. Remember that human females have two
X-chromosomes (XX), and the males have only one X and one Y chromosome.
Males receive their lone X-chromosome from their mother, and the
Y-chromosomes from their father, whereas females receives one of her
X-chromosome from her mother, and the other X from her father.
The characteristics of such inheritance are:
(a) The trait appears in almost all the generations, and the inheritance is
vertical.
(b) If the female is affected, then about half of her sons are affected.
(c) If the male is affected then all of his daughters would be affected, but
none of his sons are affected.
(d) In short, the pedigree resembles the pattern of inheritance of autosomal
dominants, except that there is no male-to-male transmission.
4. X-linked Recessive traits: These are the traits whose encoding gene is
present on the X-chromosome and its mutant allele is recessive to its
wild-type allele.
Red-green colour blindness and hemophilia, are some of its well known
examples. The characteristic features of such inheritance are:
(a) Females express the trait only when they are homozygous for the
mutant allele, whereas the males do so even when they are hemizygous
for it.
The pedigree chart would appear as the following one (Fig. 11.6):
44
LABORATORY MANUAL: BIOLOGY
(b) About half of the sons of the carrier (heterozygous for the trait) females
are affected. In case of homozygous females showing the trait, fifty
percent of her daughters and all of her sons are likely to be affected.
Therefore, the males are most affected in the population.
(c) Affected persons are related to one another through the maternal side
of their family.
(d) Any evidence of male-to-male transmission of the trait rules out the
X- linked inheritance.
5. Y-chromosome linked traits: These are the traits whose gene is present
on the Y-chromosome. The females do not have any Y-chromosome,
whereas all the males must have a Y-chromosome to be a male, and this
Y-chromosome they get from their father. Therefore, any trait linked to
the Y- chromosome must be present only in males, and certainly not in
any of the females. This is why these traits are also called male-sex limited
traits. All the sons of the affected male would express the trait whereas
none of his daughters would do so.
The pattern of the pedigree chart would be as follows (Fig 11.7):
Hypertrichosis of the ear (presence of hairs on pinna) is one most common
example of such traits.
Questions
1. How will you differentiate between autosome linked dominant and sex chromosome
linked dominant pedigree chart? Explain.
2. Discuss the differences in the patterns of autosome linked recessive and sex-
chromosome linked pedigree.
Note: Students may be asked to prepare the pedigree-chart from given data and analyse the
pattern of inheritance. The work may be done as a project.
Aim: To perform emasculation, bagging and tagging for controlled pollination
Principle: Conventional plant breeding programmes involve bringing under human control
reproductive processes that lead to seed and fruit formation. For this controlled pollination is
desirable using male and female parent having desired traits. One of the process that can be easily
brought under human control is emasculation. For this the knowledge of flower structure,
mechanism of pollination, fertilisation and physiology of flowering is essential for this. In
emasculation technique the stamens are removed before anthesis to obtain female parent and pollen
from the desired male parent is transferred on to its stigma.
Requirement: Ornamental plants/ wild plants bearing large bisexual flower, magnifying lens,
tweezers, small sharp scissors, brush, alcohol, rubber bands, paper bags, paper clips and tags
Procedure
(i) Select a flower in bud condition where antheses has not occurred. Open
the bud carefully and remove the stamens (Fig. 12.1). Mark this as
female parent plant.
Exercise 12
Fig. 12.1 Showing process of Emasculation


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LABORATORY MANUAL: BIOLOGY
Questions
1. Why is emasculation performed before anthesis?
2. What are the advantages of using a bag containing minute pores?
Fig. 12.2 Bagging of an emasculated flower
(ii) Cover the emasculated flower with a plastic
bag to protect it from undesired pollen
(Bagging) (Fig. 12.2). The bag should be
held securely in place with a paper clip/
string/ thread. Select the size of bag in
accordance with the flower size. Bags
must be transparent with minute pores.
(iii) Bring into physical contact anthers of a
desired male plant containing mature
pollen grains with the stigmatic surface
of emasculated female flower (Fig. 12.3).
Use tweezers/ brush if necessary to dust
the stigmatic surface with pollen.
(iv) Cover the pollinated flower again with the
bag immediately. For identification, label
the femal e parent (Taggi ng). Each
pollinated flower should bear a label
containing the name of the seed parent,
the letter X (to signify a cross), the name
of the pollen parent, and the date on which
the cross was effected.
Fig. 12.3 Showing cross pollination on an
emasculated flower


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Aim: Staining of nucleic acid by acetocarmine
Principle: Acetocarmine combines with nucleic acid present in the nuclei of cells to form a deep
red conjugate.
Requirements: Onion bulb, onion root tips, 2 to 4% acetocarmine/ acetoorcein stains, slide
and coverslips, brush/ needle, pair of fine scissors, filter paper and microscope
Procedure
(i) Peel off epidermis from the fleshy leaf of onion and put it on a slide.
Add a few drops of water over it to avoid desication.
(ii) Cut out a small piece (about 0.5 cm size) of the epidermal peel and
discard the remaining portion.
(iii) Wipe out the water with a filter paper.
(iv) Put 2 drops of acetocarmine on the epidermal peel and heat gently on
a spirit lamp.
(v) Apply a coverslip over the peel avoiding air bubbles and wrinkles of
the material.
(vi) Wipe out the excess stain with help of blotting paper.
(vii) Examine the material under low magnification of a microscope.
Observation
Record your observations with regard to shape of cell, the number of nuclei
and their position in the cell. Draw a diagram based on your preparation
and label its parts.
Discussion
Nuclei in cells are extremely rich in nucleic acid which exist in a conjugated
form with protein to form nucleoproteinous structures, called chromatin
fibres/ chromosomes.
Exercise 13


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LABORATORY MANUAL: BIOLOGY
Questions
1. What are the building blocks of the nucleic acid?
2. What is DNA and how is it different from RNA?
3. Name different nitrogenous bases present in the nucleic acid.


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Aim: To identify common disease-causing organisms and the symptoms of the diseases
Principle: There are quite a large number of organisms that are parasitic/ pathogenic to humans.
These organisms substantialy damage the human body and cause diseases, which may even be fatal
sometimes. These organisms exhibit characteristic features in their external morphology. Symptoms
of the diseases caused by them are also specific.
Requirement: Preserved specimens/ permanent slides/ photographs of Ascaris, Entamoeba,
Plasmodium, Ring-worm fungus and compound microscope
Procedure
Observe the preserved specimens/ slides/ photographs and note down the
features in the practical record book. Take care to observe all the minute
details and draw labelled diagrams of the pathogens.
Exercise 14
Observation
A. Entamoeba
Observe the following features of the parasite in the slide or photograph:
(i) It is unicellular.
(ii) Shape of the cel l i s i rregul ar due to
pseudopodia.
(iii) A single nucleus is present eccentrically in
the cell.
(iv) *In the nucleus a peripheral ring of granule
of nucleoprotein and central karyosome are
observed. Rest of the space in the nucleus
looks empty (Fig. 14.1).
(v) A few food vacuoles may be seen in the
cytoplasm. Contractile vacuoles are absent.
(vi) *Mature quadrinucleated cysts may be
present.
Fig.14.1 An Entamoeba


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LABORATORY MANUAL: BIOLOGY
Note: Entamoeba is an intestinal parasite in humans and causes amoebic
dysentery. The symptoms of the disease are frequent loose, mucus filled
watery stools, abdominal pain and spasms.
Systematic position
Phylum Protozoa
Class Rhizopoda
Type Entamoeba histolytica
* Distinctive feature of the pathogen
B. Plasmodium vivax
(i) It is an intracellular endoparasite seen easily within the RBC of the
infected person.
(ii) It is unicellular.
(iii) The most diagnostic stage of the parasite is "signet ring" stage in the
erythrocytes, wi thi n whi ch i t appears as a rounded
body (Fig. 14.2).
(iv) It has a big vacuole inside, and the cytoplasm is accumulated at one
place containing the nucleus. Because of the above mentioned features,
the parasite appears as a ring.
Search the stage in the blood film slide, find the signet-ring stage, and
draw its labeled diagram.
Note: It is a protozoan parasite causing malaria in humans. When an infected
female anopheles mosquito bites a healthy person, it injects the infective
stage, sporozoite, into the peripheral blood vessels. The infective stage
undergoes several rounds of multiplication in liver and erythrocytes.
Symptoms: Intermittent high fever with chills followed by profuse sweating
at an interval of alternate days.
Systematic position
Phylum Protozoa
Class Sporozoa
Type Plasmodium vivax


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51
EXERCISE 14
C. Ascaris
The external features of round worm are as follows:
(i) Body long (20 to 40 cm), cylindrical (5 to 6 mm
diameter) with no segmentation (Fig. 14.3).
(ii) Sexes are separate; the females are longer than
the males.
(iii) Both the ends are pointed; posterior end of male
is ventrally curved.
(iv) Mouth is situated at the anterior end, and is
surrounded by three lips, one present mid-
dorsal l y and rest two l i ps are si tuated
ventrolaterally (for viewing these lips a magnifying
lens is needed).
(v) Single longitudinal lines are present on the dorsal,
ventral and on the two lateral sides, all along the
length of the body. Out of these the lateral lines
are comparatively more distinct than the others
lines.
(vi) Excretory pore is present on the ventral surface
slightly behind the anterior end.
(vii) In addition to the ventrally curved posterior tip,
the male worm has a pair of penial spicules very
close to the cloacal opening.
(viii) In case of female specimen a female genital
aperture is present mid-ventrally about one third
distance from the anterior end.
Systematic position
Phylum Aschelminthes
Class Nematoda
Type Ascaris lumbricoides
Note: Round worm or Ascaris is one of the common parasite found in the
intestine of human beings.
Symptoms: (a) Irregular bowel, (b) Occasional vomiting, (c) Anaemia
Fig.14.3 Ascaris (a) Female (b) Male
Female genital
aperture
Mouth
Penial spicule
(a)
(b)


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LABORATORY MANUAL: BIOLOGY
Trichophyton (Ringworm fungus)
It is a fungus that feeds on keratin of the skin of human beings. The features
as observed under the microscope are:
1. Texture of hyphae is waxy, glabrous to cotton like.
2. Unstained hyphae are white, yellowish brown to reddish brown in colour.
Systematic position
Kingdom Fungi
Class Deuteromycetes
Type Trichophyton rubrum
Symptoms
Ringworm is a contagious fungal infection of the skin. Infected area of skin
is itchy, red, raised, scaly patches (with sharply defined edges). It is more
red on the periphery than in the center creating a ring like appearance.


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Aim: To study the texture of soil samples
Principle: Texture is one of the most important physical properties of soil. The soil texture is based
upon division of the size of soil particles into three size fractions viz., Sand (20.05mm average
particle diameter), Silt (0.050.002mm) and Clay ( less then 0.002mm). If one of these fractions
dominates the properties of a soil, the name of that fraction is included in the name of the texture.
A soil which has all of these fractions in nearly equal proportion is called a loamsoil.
The four termssand, silt, clay and
loam are combined in various ways to name
12 different textural classes. The 12 textural
classes and the percentages of sand, silt and
clay fractions that are included in each are
shown in textural triangle (Fig. 15.1).
Texture affects several physicochemical
properties of soil like density, capillary and
non-capillary pore spaces, water holding
capacity, aeration, temperature and also the
root penetration.
Requirement: Oven/ stove dried soil
samples, balance, weights, mechanical sieve
set and blotting sheets/ old newspapers
Procedure
Three methods are suggested here. Any one of these may be followed.
Method I
(i) Collect about 300500g of soil from two different locations. Label them as sample A and B.
(ii) Dry the samples in an oven, or stove or in sun to remove the soil moisture (capillary and
bound water).
(iii) Select the 3 sieves of different mesh sizes (2mm, 0.05mm and 0.002mm). Arrange them in
a collecting chamber as shown in Fig. 15.2.
(iv) Place 200g of the soil in the Ist sieve (sieve of 2mm mesh) and close the lid. To sieve the soil,
shake the set manually for 510 minutes and collect the three soil fractions.
Exercise 15
Fig.15.1 Soil Textural Triangle


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54
LABORATORY MANUAL: BIOLOGY
Soil sample Percentage (%) Texture class
Sand Silt Clay
A
B
Note for Teachers: The sieve sets contain a number and an abbreviation BSS/ ASTM/
ISS on each sieve. In the given table (Table No. 15.1) the corresponding aperture size of
the sieves is listed. For example, BSS 30 sieve aperture size will be 500 microns.
Fig.15.2 Si eve set
2mm mesh
0.05mm mesh
0.002mm mesh
Collecting chamber
Lid
the precentage lines of silt run parallel to the clay side of the triangle and, (iii)
perentage lines of sand run parallel to the silt silde of the triangle. In reading
the textural triangle, any two particle fractions will locate the textural class at
the point where these two intersect.
(v) Weigh the soil fractions viz-sand, silt and clay
collected in the 3 compartments
Wt of soil sample taken .........g
Wt of sand fraction .........g
Wt of silt fraction .........g
Wt of clay fraction .........g
The weight of three fractions should be equal to the
total weight of sample taken for analysis.
Observations
Calculate the percentages of the various soil fractions
and tabulate:
Calculate the percentages of sand, silt and clay
fractions.
Use the textural triangle now. Note that the three
sides of the textural triangle represent 0 to 100% of sand,
silt and clay respectively. Note that (i) the percentage
lines for clay run paralled to the base line of sand, (ii)


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EXERCISE 15
Appendix 1
Table 15.1 Mesh No. and the corresponding Aperture size
Sl. No. BSS Mesh ASTM Mesh ISS Mesh Aperture
1 4 5 480 4.75 mm
2 5 6 340 3.35 mm
3 6 7 280 2.80 mm
4 7 8 240 2.36 mm
5 8 10 200 2.00 mm
6 10 12 170 1.70 mm
7 12 14 140 1.40 mm
8 14 16 120 1.18 mm
9 16 18 100 1.00 mm
10 18 20 85 850 micron
11 22 25 70 710 micron
12 25 30 60 600 micron
13 30 35 50 500 micron
14 36 40 40 425 micron
15 44 45 35 355 micron
16 52 50 30 300 micron
17 60 60 25 250 micron
18 72 70 20 212 micron
19 85 80 18 180 micron
20 100 100 15 150 micron
21 120 120 12 125 micron
22 150 140 10 106 micron
23 170 170 9 90 micron
24 200 200 8 75 micron
25 240 230 6 63 micron
26 300 270 5 53 micron
27 350 325 4 45 micron
28 400 380 3 38 micron
29 25 micron


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LABORATORY MANUAL: BIOLOGY
Method II
Texture by Feel
The texture of the soil sample can also be estimated by feeling it in the dry,
moist and wet states. Sand is coarse and gritty, silt feels smooth like flour
and clay is sticky and plastic. The smallest soil particles that one can see are
coarse silt. Feel the known texture samples first, then feel the unknown ones
and decide their textures.
Procedure
(i) Feel the dry soil first. Does it crumble easily or is it hard to break?
Hard soil samples contain a moderate amount of clay.
(ii) Take in your palm a lump of soil sample about the size of a one-rupee
coin and wet it to the consistency of modeling clay. Try to press it into
a ribbon between the thumb and forefinger. An alternate test is to form
a wire by rolling the wet soil until it is about 1/ 8" in diameter.
(iii) If a long wire or ribbon can be formed readily the soil is plastic and
probably contains over 40% of clay. Its texture must therefore be clay/
silty clay/ or sandy clay. If a ribbon or wire can be formed easily but
also breaks easily, the soil sample is probably a clay loam/ silty clay
loam/ or sandy clay loam. A heavy loam/ silt loam/ or sandy loam
sample may form ribbon or a wire if the moisture content is just right
but these will be still weaker than the ribbons and wires formed by the
clay loam samples.
(iv) Next determine whether sand or silt is dominant. If there is a gritty feel
without the smooth floury touch of silt, choose a texture-name that
includes the word sandy. If the smooth floury feel predominates and
there is not much gritty feel, choose one of the silty texture names.
Use the name without a prefix if neither smoothness nor grittiness
predominates (simply clay or sand or silt). Often this can best be
determined by adding more water until the soil is in a wet state.
If the soil is very sandy, you must choose between sandy loam, loamy
sand and sand. In the moist state, sandy loam samples will have some
tendency to stick together but loam sand and sand samples will not do so.
Use the wet state to determine whether a sample is sand or loamy sand.
After handling wet sand, your hands will be moist but clean loamy sand will
make the hands slightly soiled.


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EXERCISE 15
Method III
Requirements: Soil samples, balance, weights, glass rod, standard sieves of
2mm and 0.5mm mesh size, blotting sheets/ old newspapers, evaporating
dish and water
Procedure
(i) Collect 200-300g of soil samples from different sites, and dry them as
suggested previously to remove the moisture.
(ii) Sieve the sample through a 2mm sieve to remove stones, pebbles,
roots etc.
(iii) Take 100-150 g of the sieved soil sample and further sieve it through a
0.05 mm sieve to separate the sand fraction (collected in the sieve) from
silt and clay (collected on a blotting sheet). Weigh the amount of sand
fraction and silt +clay fraction.
(iv) Take a large evaporating dish (a shallow clay plate, glass trough or a
shallow iron plate) and record its weight.
(v) Add the clay and silt fraction to the dish and note the weight.
(vi) Add water to the dish leaving half an inch space empty at the top and
stir the liquid thoroughly with a glass rod taking care that the contents
do not spill out. Allow it to stand for several hours. Decant off the cloudy
supernatant liquid (clay fraction). Repeat the process three to four times
until the decanted liquid is quite clear.
(vii) Dry the silt left in the evaporating dish to dryness. Cool the dish and
weigh it.
Observation
Record your observation in the following table:
Subtract the weight of silt fraction from the weight of silt +clay fraction. The
difference will be the weight of clay decanted.
Calculate the % of sand, silt and clay fraction of the soil and express the texture.
A B
Weight of the soil sample taken
Weight of sand fraction
Weight of silt & clay fraction
Weight of silt fraction


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LABORATORY MANUAL: BIOLOGY
Questions
1. Which type of soil is better for root-penetration and better aeration?
2. Among sandy and clay soil which one has higher water holding capacity? Explain.
3. If the clay content is high, will it affect soil fertility? Explain.
4. Which type soil has poor nutrient status and high leaching?
5. What kind of plants grow in smooth texture soil? Name two plants that grow in
heavy-textured soil.
Discussion
Correlate the texture with the plants growing in the area from which the soil
sample has been collected. Discuss how the texture of soil can affect the root
penetration, tillage, soil aeration, moisture content, water holding capacity
and other aspects related to plant growth. In sandy soil the non-capillary
pore spaces will be more and the capillary pore spaces will be less. The
condition will be reverse in case of clay soil. The pore space in turn determines
water holding capacity, percolation rate, aeration, root penetration and soil
flora and fauna. Clay particles are anionic colloides and adsorb mineral
nutrients and minimise their leaching.


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Aim: To determine the water-holding capacity of soils
Principle: Water holding capacity of the soil is the amount of water retained in the capillary
spaces of the soil after the percolation of gravitational water into the deeper layers. Water holding
capacity depends upon the capillary pore spaces in the soil. Sandy soil has very low water holding
capacity, whereas clayey soils have very high water holding capacity.
Requirement: Soil samples from different sites (garden, road side, bank of river, paddy field
etc.), Gooch crucible (china clay crucible with perforated bottom), filter-paper, pestle and
mortar, petridish, beaker, glass rod, balance and blotting paper
Exercise 16
Procedure
(i) Dig a small pit about 10cm x 10cm x 10cm, Scoop 100300 g of soil
from the pit and collect it in a small polythene bag.
(ii) Remove the pebbles and large lumps from the soil sample.
(iii) Pass the soil through a coarse sieve to remove small lumps and dead
decaying leaves and twigs.
(iv) Spread the soil into a thin layer on a sheet of blotting paper or old
newspaper and sun dry it for 23 hours or dry it in a pan kept on
stove. Alternatively dry the soil sample in oven at 108
0
C for 1 hour.
(v) With the help of pestle and mortar grind the sample into fine powder.
(vi) Put a small disc of blotting paper at the base of the Gooch crucible.
Weigh the crucible along with the blotting paper and note its weight.
(vii) Transfer the soil sample into the crucible. Tap the rim of the crucible
gently several times with the help of glass rod so that soil is compactly
filled and forms a uniform layer at the top. Add more soil if necessary.
(viii) Weigh the crucible along with soil sample and note its weight.
(ix) Fill the petridish with water and place two small glass rods in it parallel
to and at a small distance from each other.
(x) Place the crucible on the two glass rods in such a manner that its
bottom is in contact with water.
(xi) Leave the set up undisturbed till water appears at the upper surface of
the soil. Wait till entire soil surface is wet.


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60
LABORATORY MANUAL: BIOLOGY
(xii) Remove the crucible and allow all the gravitational water to flow out
from the bottom. When no more water percolates, wipe the bottom dry
with the blotting paper.
(xiii) Weigh the crucible and note its weight.
Observation
Record your observation in the following table.
Calculate the % water holding capacity of the soil as follows.
Weight of crucible +blotting paper: A g
Weight of crucible +blotting paper
+soil sample before experiment: B g
Weight of dry soil: B - A=Cg
Weight of crucible +blotting paper
+wet soil sample after experiment: D g
Weight of wet soil after the experiment: D - A=Eg
Mass of water absorbed by soil: E - C=Ng
% Water holding capacity:
Sample No. Wt. of Crucible Wt. of Crucible Wt. of soil Wt. of crucible Wt. of wet Amount of % water
+ blotting +blotting paper sample +blotting paper soil (D-A) water holding
paper (A) + soil sample (B) (B-A) = (C) + wet soil (D) =E absorbed capacity
(E-C) =N
A Garden
soil
B Road
side soil
C...
D...
Tabulate your results as shown below


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61
EXERCISE 16
Questions
1. What are heavy soil and light soil?
2. Give examples of a plant seen in heavy soil and light soil.
3. How does pore space determine the % water holding capacity of soil?
4. Why is clay soil often referred to as physiologically dry soil?
5. Which type of soil is suitable for cultivation of crop plants?
6. How can water-holding capacity of soil be improved?
7. Dead decomposed organic matter is usually added in the fields before the
cultivation of crops. Apart from providing the mineral nutrients, what additional
role does organic matter play in the cultivation of crop plants?
Discussion
Compare % water holding capacity of soil collected from different habitat
conditions. The variation in water holding capacity is due to varying
proportion of sand, silt and clay in the soil of different habitats. Soil with
very high proportion of sand have very low water holding capacity due to
large pore spaces between the particles which enables the water to percolate
freely into deeper layers leaving upper layers practically dry. In clay soil,
due to very small size of the pore spaces (fine capillaries) the water is retained
in the capillary spaces as capillary water. In these soil the water does not
percolate freely. Soil with more or less equal proportion of sand, silt and clay
(loam soil) combines the properties of sand and clay and therefore has
optimum water holding capacity and optimum soil-air for root growth.


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Aim: To study the ecological adaptations in plants living in xeric and hydric conditions
Principle: Successful adjustment of plants and animals under prevailing environmental conditions
is known as adaptation. For terrestrial plants, the habitats vary from extremely dry conditions as in
deserts to extremely wet conditions as in marsh lands. For aquatic plants the habitats may vary from
deep water bodies like oceans and lakes to shallow ponds and pools. The plants are adapted to
diurnal, seasonal or annual fluctuations of the habitat conditions. For land plants the main limiting
factor is the availability of soil water whereas, for aquatic plants the main limiting factors are the
fluctuations in water level, availability of gases like CO
2
and O
2
and the light intensity. Adaptation
of land plants are primarily for conservation of available soil water, avoidance of bright sunlight
and intense heat and for aquatic plants, adaptation are for conservation of gases and efficient
utilization of available sunlight.
On the basis of availability of water, plants are classified as:
(a) Xerophytes: These are plants growing in extreme dry conditions throughout the year. For
example, plants growing in deserts (psammophytes), on rock (lithophytes) or alpine plants
growing above 14000 feet altitude.
(b)Mesophytes: These are plants growing in soils with optimum soil water conditions prevailing
for major part of the year.
(c) Hydrophytes: These are aquatic plants growing in fresh to marine water.
The morphological, anatomical and physiological attributes of terrestrial plants are different
from the aquatic plants.
Requirement: Plant specimens from xeric and hydric habitat conditions. The specimens from
xeric condition may include a few cacti, succulents (Euphorbia, Bryophyllum, Kelancho) cycas
leaves, pine needles, twigs of Acacia, Nerium, Parkinsonia, Casuarina etc. The aquatic plants:
Salvinia, Eichornia, Pistia, Hydrilla, Vallisnaria, Utricularia, Lymnophila; some reeds like Typha,
Phragmites, amphibious plants like Marsilea and halophyte like Rhizophora. Beakers, glassjars,
microscope, slide, coverslips and rajor blades
Exercise 17
Procedure
Prepare temporary stained transverse sections of leaf, stem and root of the
specimens. Study the morphological and anatomical features of the plants


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63
EXERCISE 17
collected and look for the following adaptations. Write the name of the plant
in which a particular adaptation is observed.
Observations
Record your observation in the given tables:
Xerophytes
Adaptations
1. Conservation of Water
2. Storage of Water
3. Preventi on of l oss of
water by transpiration
4. Prevention of excessive
heat
5. Efficient mechanism of
water absorption
Modifications
(Morphological/ Anatomical)
a. Leaves few or absent or represented
by spines only
b. Peti ol e modi fi ed i nto l eaf l i ke
structure
c. Stem reduced, branching sparse
d. In some cases stem flattened, leaf
l i ke, green, photosyntheti c i n
nature
Thick, fleshy and succulent
leaves as well as stem
a. Intercellular spaces reduced
b. Spongy parenchyma/ pal i sade
parenchyma present
c. Stomata on lower surface,
sunken in stomatal pits
d. Leaves needle like
e. Thick cuticle on leaf surface
a. Leaves covered with dense hairs;
b. Leaf surfaces shiny or glaborous
c. Leaf blade remains rolled during
the day
a. Long and profusely branched roots
b. Dense root hairs
c. Well developed xylem
Examples (from the
specimen collected)
------
--------
---------
-------------
-----------------


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64
LABORATORY MANUAL: BIOLOGY
Questions
1. Give three adaptive features of water hyacinth suitable to aquatic life.
2. What are the features present in plants of xeric habitat for the prevention of
loss of water?
3. What is the importance of succulent leaves and stem for a xerophytic plant?
4. Why is air stored between tissues in aquatic plants?
Hydrophytes
Adaptations
1. Buoyancy and resistance
to currents of water
2. Transpiration
3. Absorption of water
4. Gaseous circulation and
storage of air
5. Mechanical tissues
Modifications
(Morphological/ Anatomical)
a. Leaves long and cylindrical
b. Peti ol es fl exi bl e to wi thstand
currents of water and to carry the
leaf blade on the surface of water
c. Peti ol es are modi fi ed i nto ai r
pockets
d. Leaf blade pale green in colour,
finely dissected
e. Leaf blade waxy with thin cuticle
a. Stomata absent
b. Stomata present on upper surface
of leaves
a. Poorly developed roots
b. Root hairs absent
c. Roots with air pocket to help in
buoyancy
Parenchymatous tissue of stem, roots,
peti ol es and l eaves modi fi ed i nto
aeranchyma in the form of air channels
i n
a. Root
b. Stem
c. Petiole
d. Leaf
a. Poorly developed xylem
b. Poorly developed sclerenchyma
c. Sclerides present
Examples


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Aim: To study the adaptations in animals living in xeric and hydric conditions
Principle: The aquatic ecosystem exhibits adifferent pattern of abiotic factors as compared to those
in terrestrial ecosystem. The temperature of the water, penetration of sun light, the physicochemical
characteristics of water body affect the growth and survival of the biotic community. In order to over-
come the cumulatory effects of these factors, certain morphological and anatomical features, as well
as physiological processes develop in the organisms. These modifications in animals are called adaptive
features. We will study adaptations in selected animals living in aquatic and xeric condition.
Requirement: Animal specimen/ models of xeric (rat, camel, squirrel) and hydric (fish, frog,
prawn, etc.) conditions
Exercise 18
Procedure
Observe the animals provided and note down their adaptive features in the
observation table with example.
Observations
Hydric adaptations
Features
Body colour
Body contour
Locomotory
Adaptations Example
(a) On dorsal surface
(b) On ventral surface
(a) Streamlined
(b) Disappearance of neck constriction
(c) Tail enlargement
(d) Position of external nostrils
(e) Loss of external ears
(f) Position of eyes
(g) Presence of eye protecting membrane
(a) Fins or fin-like expansions of the body wall
(b) Loss of limbs
(c) Webbed feet
(For students)


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66
LABORATORY MANUAL: BIOLOGY
Integument
Mouth
Respiratory organs
Presence of dermal/ epidermal derivatives
(a) Scales
(b) Hairs
(c) Mucous glands
(d) Oil glands
(a) Position
(b) Presence of teeth
a. Upper jaw
b. Lower jaw
(a) Gills/ lungs
(b) Cutaneous
Xeric adaptations
Features
Moisture getting
Moisture Conservations
Body colour
Body contour
Skin
Limbs
Scrotum
Adaptations
(a) Preference for juices as food
(b) Hygroscopic skin
(a) Storage of water in body
(b) Avoidance of evaporation (non-perspiring)
(a) Protective mimicry
(b) Predating mimicry
(a) Position of
a. Nostrils directly upward
b. Reduction to pin-head size
(b) Position of eyes
a. Covering of eyes
b. Size
(a) Hard
(b) Spiny
(c) Poison glands
(a) Speed
(b) Long slender
(c) Padded feet
Present or Absent


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67
EXERCISE 18
Questions
1. Name the features that helps a frog for aquatic life.
2. What are the adaptations present in xeric animals for conservation of water?
Discussion
You may have noticed many features in the body of aquatic animals
which support their life. As the different aquatic bodies vary to a
great extent, there are many other adaptive features you may notice.
For example the aquatic organism in ponds, lakes, river and sea.


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Aim: To determine the pH of different water and soil samples
Principle: The pH value of a water/ soil sample can be determined by (i) indicator dye method,
(ii) electrometric method using a pH meter and (iii) colorimetric method. For routine purposes
the indicator dye method using universal pH indicator solution (containing a wide range of pH
indicator dyes) or paper strips containing the pH indicators are preferred though it is not as
accurate as the electrometric method.
Requirement: Soil or water samples A, B and C collected from different sites (for example
soils from road side, garden, humus rich sites; water samples from borewell, handpump, pond,
sewage), balance, weights, filter paper, distilled water, measuring cylinder (50 mL), droppers,
cavity tile, funnel, beakers (100 mL), funnel stand, universal pH indicator solution and
pH indicator paper (narrow range and broad range)
Exercise 19
Procedure
(i) Weigh 10 g of the soil sample A. Add 50 mL of distilled water to soil
sample to make a soil solution.
(ii) Filter the soil solution through a filter paper and collect the filtrate in a
beaker. Label it as soil solution -A.
(iii) Take a clean dry porcelain cavity tile. Place 5 drops of soil solution A
in three cavities of the tile as shown in Fig. 19.1.
Soil - A Soil - B Soil - C
Universal pH indicator
solution
pH indicator paper
(Broad range)
pH indicator paper
(Narrow range)
Fig. 19.1 Porcelain cavity tile


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69
EXERCISE 19
(iv) To the 5 drops of soil solution present in one cavity add 5 drops of
universal pH indicator solution. Note the colour developed and compare
i t wi th the col our chart gi ven on the uni versal pH i ndi cator
solution bottle.
(v) To the soil solution present in the second cavity, dip a small strip of broad
range pH indicator paper (pH 2-11). Note the colour and compare with
the colour chart given on the broad range indicator paper and get a rough
estimate of pH of the sample solution.
(vi) Choose a suitable narrow range pH indicator paper (for e.g. If the pH of
soil is determined by you as 8.0, choose a narrow range 7.0 to 9.0) and
dip a small strip of it in the soil solution present in the third cavity. Note
the colour developed and determine the pH to the nearest possible value
with the help of the colour chart.
Repeat the same steps for determining the pH of sample B and C. Follow the
same procedure for water samples collected from different sites.
Observation
Record your observations in the given table.
Discussion
Based on the pH values obtained, categorise the samples into acidic, basic,
neutral type.
Record the plant species present in the site from which the samples are collected.
Note for teachers: The colour developed should be noted against direct sun
light. Also, sometimes the soil solution colour may interfere with the readings.
Thus one has to be careful while making the observations.
pH value as determined by Soil Samples
A B C
Universal indicator solution
Broad range indicator paper
Narrow range indicator paper
Table: Measurement of pH of soil samples A, B and C


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LABORATORY MANUAL: BIOLOGY
Questions
1. What will be the pH of chalk (calcareous) soil?
2. pH measurement with indicator paper is not very accurate. Comment.
3. Water logged soils are acidic. Comment.
4. Why are soil around mineral mining areas acidic?


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Aim: To study turbidity of water samples
Principle: Various characters that control the quality of water are taste, smell, colour, amount of
dissolved nutrients, dissolved O
2
and CO
2
, pH and different types of plants and animals and their
density. Turbidity of the water body determines the depth upto which light can penetrate and thus
affects the distribution and photosynthesis of phytoplanktons and macrophytes. More turbid the
water body less is the thickness of its photic zone.
In polluted water bodies turbidity is due to:
1. Effluents: A water body which receives domestic sewage, run off from adjacent agricultural
fields and liquid wastes from nearby small and large industries remains turbid.
2. Planktons: A water body may be turbid due to very high density of phytoplanktons and
zooplanktons, especially when the water body is rich in nutrients.
I. Secchis Disc method
Requirement: Secchis Disc, rope of moderate thickness, meter rod, black and white
paints; paint brush. Prepare aSecchi's disc by taking an iron disc of about 6 inches diameter, to
which aweight is attached in the centre on one side and an iron hook on the other side. Tie aplastic
rope of sufficient length to the hook. Divide the upper surface of the disc into 4 equal segments and
paint two of these white and the other two segments black in such a way that black and white
segments alternate with each other (Fig. 20.1).
Exercise 20
Procedure
(i) Visit a nearby pond.
(ii) Reach to the center of the pond in a small boat.
(iii) Slowly immerse the Secchi disc into water vertically holding
the rope tightly in the hand till the black and white segments
of the disc just begin to disappear. On reaching to a
particular depth, the disc becomes completely invisible.
Mark the length of the rope when the disc just disappears
(say A cm).
Fig. 20.1 A Secchis Disc


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LABORATORY MANUAL: BIOLOGY
(iv) Slowly pull up the disc and find out the length of the rope where the
black and white segments of the disc just reappear (say B cm).
(v) Find out the mean length (X) of the rope by the following method.
(vi) Repeat the process at different sites of the pond.
Water body Depth at which Disc Depth at which Depth of Photic
disappears (A cm) disc reappears (B cm) zone
Pond Site 1
Site 2
Site 3
-
-
Tabulate the results in the given table
Observations
The value X represents the depth of the photic zone upto which sunlight
penetrates in the water body and photosynthesis takes place.
Discussion
Greater the value of 'X' less turbid is the water. In crystal clear deep lakes, the
value of 'X' will be very high indicating, thereby, that the water body does not
have large quantities of flocculating silt or organic matter residues. This may
be due to no discharge of effluents or domestic sewage into the water body.
The high clarity of water is also an indication of very less density of phyto and
zooplanktons. These water bodies are called as non-productive or oligotrophic,
while highly turbid water bodies are eutrophic in nature.
Precautions
Students are advised to perform this experiment under the strict supervision
of teacher to prevent incidents due to drowning.


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EXERCISE 20
II. Measurement of turbidity using measuring cylinder
Requirements: Water samples from different sources, three measuring
cylinders (500mL) of the same height.
Procedure
(i) Collect about 2 liters each water samples from different sources.
(ii) Transfer 500ml of water sample in the measuring cylinders of same
volume and height.
(iii) Mark the three cylinders A, B and C and leave them undisturbed
overnight.
Observations
Observe the amount of sediment settled at the bottom of each cylinder and
also note whether the water above the sediment is still turbid.
Discussion
Do all the samples show same amount of sediments?
Which sample shows maximum sedimentation and correlate it with the
source of the sample?
Find out whether in all the cylinders, water above the sediment is clear or
turbid. Explain with reasons.
Draw conclusions on the basis of the observations.
Water sample Thickness of sediment Clarity of waterturbid/
semiturbid/ clear
'A'
'B'
'C'
Record your observations in the following table:


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LABORATORY MANUAL: BIOLOGY
Questions
1. Is turbid water fit for drinking? Explain.
2. Why is the penetration of sunlight in any water body important?
3. Green plants are seen only in photic zone. Comment.
4. It is a common practice to use alum for clearing turbid waters. Explain.
5. Turbidity of water body varies with season. Comment.


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Aim: To analyse living organisms in water samples
Principle: The productivity and the trophic status of a water body is determined by assessing the
number and type of organisms (micro as well as macro) present in the water body. Water body with
very high density of phytoplankton per unit area is a productive water body. Such water bodies are
usually turbid and have high amounts of nutrients and dissolved oxygen. These water bodies support
fairly large number of organisms of different trophic levels. This is in contrast to non-productive
water bodies, which have very low density of organisms per unit area, fairly transparent waters with
low mineral concentration and dissolved oxygen and also fewer trophic levels. The status of health
of a water body can be determined by analysing water samples for the number and type of organisms
present in it at a given time. Such assays also help us to find out whether a water body is polluted as
some of the organisms are strong indicators of water pollution.
Requirement: Water samples from different water bodies (lake, pond, river etc.), beakers,
a few vials or small test tubes, slides and cover slips, watch glasses, dropper, compound
microscope and 5% FAA (Formalin Aceto Alcohol 5:5:90:Formalin: Acetic acid: Ethanol) as
preservative
Exercise 21
Procedure
(i) Collect about a liter of water sample from nearby water body (pond lake,
reservoir, river etc).
(ii) Add about 5 ml of FAA to fix and preserve the living organisms present in
each sample at the place of collection.
(iii) In the laboratory, transfer the water sample into a measuring cylinder of
one litre capacity. Label each water sample to indicate the site from which
the water sample has been collected.
(iv) Leave the water samples undisturbed for 48-72 hours.
(v) Decant off the clear water, leaving concentrated sediment at the bottom.
(vi) Transfer the sediment into a vial or a small test tube. Cork and label each
vial for future use.
(vii) With the help of a dropper, transfer a few drops of sediment liquid from a
vial into a watch glass. Dilute the sediment with water if the sediment is
highly concentrated.


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LABORATORY MANUAL: BIOLOGY
Questions
1. Why do you find few organisms in polluted water? Explain.
2. Why is FAA (Formaline Aceto Alcohol) added after collecting the water sample?
3. Name at least one phytoplankton and zooplankton commonly found in polluted
water.
(viii) With the help of a dropper transfer a drop of water from the watch glass
on the center of a slide and mount it. Blot the excess water using blotting
paper.
(ix) Prepare a few more slides of each water sample in the same way.
(x) Observe each slide, first under lower magnification and then under higher
magnification.
Observations
1. Record the different types of organisms present.
2. Count the number of organisms under each field of microscope.
3. Some of the commonly found organisms of water bodies are given in
Annexure 2.
Discussion
Prepare a list of organisms observed in each water sample and make an
assessment of type and density of different organisms in each water sample.
Polluted waters may contain very few types of organisms but in very high
density. The non-polluted waters will have large variety of organisms in
low density.


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EXERCISE 21
Cosmarium
Desmickum
Spirogyra
Stigeoelomium
Draparnaldiopsis
Moubeotia
Nitella
Dinobryon
Euglena
Annexure 2


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LABORATORY MANUAL: BIOLOGY
Gymnodinum
Ceratium
Peridinium
Calothrix
Anabaena
Cylindespermum
Rivularia
Scytonema
Fischerella
Gleotrichia
Cyanomonas


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EXERCISE 21
Volvox
Chlamydomonas
Gol enki ni a
Coelastrum
Closterium
Pedrastrum
Hydrodictyon
Chlorella
Ankistrodesmus
Staurastrum
Scenedesmus


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LABORATORY MANUAL: BIOLOGY
Microcystis
Synechococcus
Nostoc
Spirulina
Lyngbya
Phormidium
Synechocystis
Oscillatoria
Gloeocapsa
Mel os i r a
Chaetocer os


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EXERCISE 21
Rhi zos el i na
Bi ddul phi a
Baci l l ar i a
Amphor a
Ni tzs chi a
Centronella
Synedra


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LABORATORY MANUAL: BIOLOGY
Pleurosigma
Lauderia
Skeletonema
Coscinodiseus
Somphonema
Navicula
Fragilaria
Cocconeis
Asterionella


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Aim: To determine the amount of Suspended Particulate Matter (SPM) in air at different sites
in a city
Principle: Environmental pollution is the unfavorable alteration of our surroundings wholly or
largely as a by-product of man's action through direct or indirect effects of changes in energy
patterns, radiation levels, chemical and physical constitutions of environment and abundance of
organisms. Substances that cause pollution to the environment are called pollutants. They are the
residues of things that man makes, uses and throws away. These residues pollute soil, water and air.
The atmosphere in highly populated area is very rich in dust, smoke and SPM all due to vehicular
exhausts and industrial emission.
Requirement: A few freshly cut broad leaves, Vaseline, laboratory balance, weights, brush,
paper clips and twine thread
Exercise 22
Procedure
This experiment is an outdoor activity and may be
conducted by assigning 23 students into a group.
(i) Collect a few locally available broad leaves from a
nearby tree plant (Canna, Peepal, etc.).
(ii) Wash the leaves gently in running water to remove
any dust settled on their surfaces.
(iii) Blot dry the surface area of the leaves. To calculate
the area of the leaf, trace the outline of the leaf on
graph paper (Fig 22.1). Within the traced area
calculate the total number of full squares, 1/ 2, 1/ 3
and 2/ 3 squares and individual small squares. Add
all the squares to get the total leaf area. Multiply
their value with two to obtain total area of both the
surfaces.
(iv) Take 810 feet long twine thread and tie five leaves
leaving a foot distance in between. Apply an
extremely thin layer of vaseline on both surfaces of
each leaf. Make a bundle of these leaves and pack
Fig. 22.1Calculating the area of a
leaf on a graph paper


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LABORATORY MANUAL: BIOLOGY
them in polythene bags. Ensure that the outer surface of polythene
bag does not have any vaseline sticking on it.
(v) Make three such bundles of smeared leaves, each bundle containing 5
leaves.
(vi) Mark bundles as A, B and C and carefully weigh each bundle of leaves
along with the polythene bags.
(vii) Select three spots (X, Y and Z) near by your school. Spots selected
should be in a manner that spot 'X' has very heavy vehicular traffic,
the spot Y has moderate traffic and spot 'Z' has little or no vehicular
traffic. At spot 'X' expose each leaf of bundle 'A' by stretching the
attached thread and tie the two ends to two poles or branches of trees
preferably at 10 feet height above ground. Keep leaves exposed for
about two hours.
(viii) After exposure at spot 'X', collect the leaves and carefully re-bundle
exposed leaves and place them along with the string in the polythene
cover 'A'.
Site Leaf bundle Weight of leaves (g) Weight of Total leaf area
sample suspended (cm
2
) of five
particle (W
2
-W
1
) leaves
Before After
exposure (W
1
) exposure (W
2
)
X 'A'
Y 'B'
Z 'C'
Record your findings in the following table:
(ix) Repeat the same process at spot 'Y' and 'Z' exposing leaves of 'B' and
'C' bundles respectively.
(x) At the end of the experiment, return back to the laboratory. Reweigh
each bundle of exposed leaves along with their respective polythene
cover.
- Calculate the amount of suspended particles deposited in mg cm
2
of
leaf at each spot.
- Compare the results of three different spots and interpret.
Since the weight of suspended particles will be in milligrams or even less
it is advised to use a very sensitive laboratory balance.


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Aim: To study plant population density by quadrat method
Principle: Density represents the numerical strength of a certain plant species in the community
per unit area. The number of individuals of the species in any unit area is its density. The unit area
may be as small as 5 square cm to as large as 10 square metre depending on the size and nature of
the plant community under study. For herbaceous vegetation a metre square quadrat is normally
used. Density which gives an idea of degree of competition is calculated as follows.
Total number of individual(s) of the species in all the sampling unit (S)
Density=
Total number of sampling units studied (Q)
The value thus obtained is then expressed as number of individuals per unit area. When the
measured unit area is divided by the number of individuals the average area occupied by each
individual is obtained.
Requirement: Cotton/ nylon thread (five meters), 4 nails and a hammer
Exercise 23
Procedure
(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
vegetation is not damaged while laying the quadrat.
(ii) List the names of the plant species seen in the quadrat (if the name is
not known mark these as species A or B etc., and the same species if
seen in other quadrats assign the same alphabet).
(iii) Count the number of individuals of each species present in the quadrat
and record the data as shown in the table.
(iv) Similarly make nine more quadrats randomly in the site of study and
record the names and number of individuals of each species.
Observations
Record the total number of species seen in the ten quadrats. This will give
an idea about the composition of the vegetation.
There will be difference in the species composition in the quadrats made in
shady areas, exposed areas with bright sunlight, dry or wet areas etc.


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LABORATORY MANUAL: BIOLOGY
Discussion
Plants growing together exhibit mutual relationships among themselves and
also with the environment. Such a group of plants in an area represent a
community. The number of individuals of a species varies from place to
place, making it necessary to take many random sample areas for reliable
results. Density values are significant because they show relative importance
of each species. With increasing density the competition stress increases
and the same is reflected in poor growth and lower reproductive capacity of
the species. Data on population density are often very essential in measuring
the effects of reseeding, burning, spraying and successional changes.
Discuss the vegetation composition of the area (herbs/ shrubs) and
comment on the dominant component species.
Questions
1. What factors influence the population density?
2. What is the significance of quadrat method?
3. What conclusion can be drawn if density of a plant species is low?
Plant Quadrats employed in study & no. of Total No. of Total no. Density (D)
Species individuals in each quadrat individuals (S) of Quadrats
studied (Q)
I II III IV V VI VII VIII IX X
A 2 5 7 10 3 27 10 27/ 10 =2.7
Z 1 2 4 8 3 2 20 10 20/ 10 =2.0
Table 23.1: Density studies of the given vegetation


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Aim: To study plant population frequency by quadrat method
Principle: Frequency is concerned with the degree of uniformity of the occurrence of individuals
of a species within a plant community. It is measured by noting the presence of a species in random
sample areas (quadrats) which are distributed as widely as possible throughout the area of study.
Frequency is the number of sampling units (as %) in which a particular species (A) occurs. The
frequency of each species (sps. A or sps. B or sps. X etc) is expressed in percentage and is calculated
as follows.
% Frequency or
Number of sampling units (quadrats) in which the species occurs
=
Frequency Index Total number of sampling units (quadrats) employed for the study
Requirements: Cotton/ nylon thread of 5 metres, 4 nails and a hammer
Exercise 24
Procedure
(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
vegetation is not damaged while laying the quadrat.
(ii) List the names of the plant species seen in the quadrat (if the name is not
known mark these as species A or B etc. and if the same species is seen in
other quadrats assign the same alphabet)
(iii) Similarly lay nine more quadrats randomly in the site of study and record
the names of individuals of each species.
(iv) Calculate the percentage frequency of occurrence using the formula given.
Observations
Record the total number of species seen in the ten quadrats. This will give an
idea about the composition of the vegetation.
There will be difference in the species composition in the quadrats made in
shady areas, exposed areas with bright sunlight, dry or wet areas etc.
Observe that the frequency of occurrence is not the same for all species.


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LABORATORY MANUAL: BIOLOGY
Discussion
Variation in distribution of a species is caused by factors like soil conditions,
quantity and dispersal of gemmules, vegetative propagation, grazing, predation,
diseases and other biotic activities. Also frequency values differ in different
communities. They are influenced by micro-habitat conditions, topography,
soil and many other environmental characteristics. Thus unless frequency is
not correlated with other characters such as density, frequency alone does not
give correct idea of the distribution of a species.
Frequency determinations by means of sample areas are often needed in
order to check general impressions about the relative values of species. Many
species having low cover or population density also rate low in frequency, but
some may have high frequency because of their uniform distribution. Usually
if the cover and population density are high, the frequency will be high. The
plants with high frequency are wide in distribution.
Questions
1. If frequency of a plant is high, what will be your interpretation?
2. Can many micro-habitat in an area affect frequency of a species? Comment.
Plant Number of quadrats employed No. of quadrats Percentage
Species in the study (Q) in which the of frequency
species is present (N) F=N/ Q X 100
I II III IV V VI VII VIII IX X
A 5 5/ 10 100 = 50%
B 1 1/ 10 100 = 10%
C 4 4/ 10 100 = 40%
Table 24.1: Frequency studies for the given vegetation


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Aim: Study of homologous and analogous organs in plants and animals
Principle: In plants and animals there are several organs or parts thereof, apparently alike in
their function and appearance, but markedly different from each other in their origin and
anatomical structure. These organs are called analogous organs, and the seeming similarity
among them is the result of convergence, that is, adaptation to similar habitat and identical
ecological niche.
On the other hand, there are organs or parts thereof, which apparently are quite dissimilar
to each other in appearance and perform different functions, but have the same origin and
anatomy. The differences in their function and also in their appearances are the result of
divergence, due to adaptive radiation to different habit, habitat and ecological niche. These
organs are called homologous organs.
Requirement: Plant specimens showing tendrils, thorns, etc., as given in the text or any other
locally available plants, a plant with normal stem, potato and onion bulb, prickly pear, specimens
of phylloclade, cladode, wings of bird, cockroach and bat, and cervical, thoracic and lumbar
vertebrae of a mammal/ lizard
Exercise 25
Observations
1. Homologous Organs in Plants
(i) Tendrils of passion flower and thorns of pomegranate
Tendrils of passion
fruit and thorns of
pomegranate are
structural l y and
functionally different
but they have similar
origin i.e. they arise
from axi l l ary bud
(Fig. 25.1a & b).
Fig. 25.1(a) Tendrils of passion fruit (b) Thorns of pomegranate
(a)
(b)
Tendril
Thorn


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LABORATORY MANUAL: BIOLOGY
(ii) Tendrils of Vi ti s and thorns of
Cari ssa
Tendrils of Vitis and thorns of
Carissa originate from the terminal
bud, but they are functionally
different (Fig. 25.2 a & b).
(iii) Tendrils of baloon vine
(Cardi ospermum) and bulbils of
Agave.
Both are modifications of floral bud,
but they perform different functions.
Tendrils help in climbing but bulbils
are meant for reproducti on
(Fig. 25.3 a & b).
(iv) Scale leaves of onion and spines
of prickly pear (Opunti a)
Both the scale leaves and spines are
modifications of leaves but are
structural l y and functi onal l y
different. Scale leaves of onion are
thick and fleshy and store food. On
the other hand spines of cactus are
defensive organs (Fig. 25.4 a & b).
2. Analogous Organs in Plants
(i) Stem tendrils and leaf tendrils
All tendrils are analogous with one
another, being structurally and
functionally similar, irrespective of
their origin.
Exampl e: Tendri l s of pea and
tendrils of Vitis. Tendrils of pea are
modification of leaf and in Vitis it is
the modification of terminal bud
(Fig. 25.5 a & b).
Fig. 25.2 (a) Tendrils of Vitis (b) Thorns of Carissa
(a) (b)
Tendril
Thorns
Fig. 25.3 (a) Tendrils of baloon vine (b) Bulbils of Agave
(a) (b)
Tendril
Fig. 25.5(a) Tendrils of pea (b) Tendrils of Vitis
(a)
(b)
Tendril
Tendril
Fig. 25.4 (a) Scale leaves of onion (b) Spines of cactus
Spines
(a)
(b)


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EXERCISE 25
(ii) Thorns and spines
Thorns and spines are analogous structures being
defensive in function. Thorns are modifications of
axi l l ary or termi nal buds, and spi nes are
modifications of leaves.
e.g: Thorns of pomegranate and spi nes of
prickly pear.
(iii) Modified underground stems and modified roots
Modi fi ed stems (rhi zome, corm, tuber) are
analogous to modified roots (carrot, radish) as they
perform similar function of storage of food but their
origin is different. Rhizome of ginger, potato tuber,
Colocasia are stems and beetroot, radish etc. are
roots. (Fig. 25.6 a & b)
(iv) Phylloclade, cladode and leaves
They perform the same functi on i .e. they
photosynthesise but phylloclade and cladode are
modifications of stem. Phylloclade of Opuntia,
Parkinsonia, Asparagus and leaves of any local
pl ant l i ke mango are anal ogous organs.
(Fig. 25.7 a & b)
3. Homologous Organs in Animals
(i) Wings of birds, and forelimb of mammals/ reptiles/
frog: All have the same bony elements (humerus radio-
ulna, carpals, metacarpals and phalanges), but
perform different (flying in birds, for holding or walking
etc. in other) functions. (Fig. 25.8 a & b)
4. Analogous Organs in Animals
(i) Wings of dragonfly/ cockroach/ butterfly and of
birds. (Fig. 25.9 a & b)
(ii) Mandible of cockroach and mandible (lower jaw) of
a vertebrate. (Fig. 25.10 c & d)
Fig. 25.7 (a) Phylloclade (b) Cladode of
ruscus
(a) (b)
Questions
1. Suggest examples of homologous and analogous organs other than what are
given in the manual.
2. Why are stem and leaf tendrils considered as analogous organs?
Fig. 25.6(a) Modified root of carrot
(b) Rhizome of ginger
(a) (b)
Note: Students and teachers are suggested to discuss
more examples.
Spine
Fig. 25.8Fore limb of (a) human (b) bat
(a) (b)
Fig. 25.9 Wing of (a) dragonfly (b) bird
(a) (b)
Fig. 25.10 Mandible of (a) cockroach
(b) rabbit
(a) (b)


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Investigations are more open-ended than practical exercises involving a search to understand
the unknown and begins with a question or a hypothesis. You are not instructed exactly what
to do, but are given only general guidance. These give you more opportunity to plan your
work. For example, you might investigate what traits you and your classmates inherit from
your parents and forefathers (both maternal and paternal).
Projects are even more open-ended than investigations. These are practical investigations
carried out by an individual or a group of students. Projects are largely your own initiative. It
also requires evaluation of your findings, redefining ideas and designing further investigations.
This may lead to evidence that enables answering the question posed at the outset. Some of
these projects would take about few hours to complete. Other may take few weeks. Some are
laboratory based, others involve fieldwork. Many could be carried out at home.
Investigatory projects are part of obligatory assignment involving purely experimental
procedures so that you report on, duplicate, or adapt something that someone else has already
discovered. It may involve some other form of investigation also. For example, you may undertake
to investigate the richness and patterns of biodiversity (flora and fauna) in your school campus
and prepare a mural of it or to investigate the effects of physical fitness on your pulse rate.
Investigatory Project Work
Choosing an Investigatory Project
You may be guided by your teacher for your choice of topic. The more original
or new the project is, the better it would be. But it must be realistic in terms of
the time available and at a level attained in the higher secondary biology.
You must review the available literature to find out what type of work has
been done. This will help you to reject some of the alternatives, and possibly
cause you to modify others. It may also be the source of new ideas.
By doing these investigatory projects you will gain experience of research
besides providing opportunity for learning skills such as photography,
electronics, etc.
Identifying the Objectives of the Project
Having identified a possible project, you should be able to identify and list the
tentative objectives you hope to attain by completing that investigation. For example,


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93
INVESTIGATORY PROJ ECT WORK
Suppose your project involves studying the biodiversity of birds in your
district/ state, examine the data in the light of some questions (say, how do
the birds in Rajasthan differ from those in Assam or Bihar?) your
investigation might attempt to answer.
Suppose your project involves investigating leaf mosaics revealing the
complexity of the growth correlations that lead to efficient light interception,
suggest also the factors that might affect this type of study.
Keep the aim of your project simple. Investigate only one factor at a time
and never allow yourself to be side-tracked. Remember that time may be too
short for follow-up and any fascinating secondary aspects that you may come
across.
Designing Projects
Having established the objectives of your chosen project, you must have an
experimental design. This will allow you to collect the data you need in a
scientific way to test the hypothesis. For example, if your project involves
investigating the hypothesis that stale milk contains more bacteria than fresh
milk, devise the procedure you would adopt to carry out your investigation.
Planning Investigations
Having decided your topic for scientific investigation, you should give careful
thought to the plan of your investigation in some detail. These may include
What hypothesis can you make?
How can you ensure that the experimental tests and measurement you
carry out are accurate and reliable?
What controls do you need?
How many variables are you investigating? Correctly identify key variables
as independent and dependent.
Are your variables discrete or continuous?
Identify appropriate control variable for fair test.
How many repeat observations or samples will you require?
What instruments/ equipment or techniques will you use to obtain relevant
information? Identify suitable materials and equipment to be used.
If your investigation requires the use of a questionnaire, design and
standardise before implementation.
Is your intended procedure safe and ethically permitted, i.e., taking care of
the distress or suffering of living organisms and damage to the environment?
How will you collect your data?


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94
LABORATORY MANUAL: BIOLOGY
How do you plan to analyse your results? Would you employ statistical or
other methods? Are scale range, interval, number of values chosen are
adequate and reasonable ?
Executing the Project
Following planning, a brief description of the expected procedures has to be
approved in advance by the teacher. Having decided what controls you need
to use, list the components of your experiment and decide what quantities of
substances to use, how to set the experiment. You should also decide what
type of readings or measurements you are going to make, how often and how
many. Note the source of error, if any, that you come across.
Handle instruments and equipments appropriately to give accuracy.
Repeat measurement.
Keep proper controls and the variables constant.
Reporting/Writing of Project
A format, such as given below, can be followed.
(i) Title of the investigatory project: Write the title of the project, for
example, Inheritance pattern of eye colour.
(ii) Objectives: Express as clearly as possible the effect of one variable that
the experiment is designed to investigate.
(iii) Materials needed: This might be just a list, or a diagram if a particular
piece of apparatus was used.
(iv) Method: Describe the procedure stepwise including the precautions
taken, if any.
(v) Result: A suitable chart or table for recording and organising your
readings or measurements should be made out before you start the
experiment.
(vi) Analysis and interpretation: Observation data are factual, and may
not be as expected by you.
(vii) Discussion: Discuss briefly the implication of your results and suggest
extensions of any kind that can be undertaken.
(viii) Conclusion: In view of the results obtained and related work done on
the topic of the project, write conclusion briefly.
(ix) References: Any work related to the project which you have come across
through books/ articles or any other source should be written as reference,
for example: Michael Michalco (2001), Cracking Creativity, Berkeley, Ten
Speed Press.
This write up is meant to train the students in scientific methods. In other
words, it accentuates the spirit of enquiry and investigation in young minds.


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95
INVESTIGATORY PROJ ECT WORK
The operational aspects of doing a project include choosing a hypothesis or
problem to be investigated, collecting data in a designed manner, analysing
the data in a scientific way, drawing conclusions which are justified and
discussing the results in the light of known knowledge and bringing out its
importance. Finally it includes the scientific way of communicating the findings.
While your discovery during the investigatory project may not merit a Nobel
Prize it may help you discover something, a fact or relationship that was
unknown to you and that was not recorded in any book available to you.
Scientists refer to this as an independent discovery. Your investigation will
certainly give a sample of the thrill of discovery.
Following are pages on procedural guideline about a few suggestive
investigatory project work.
1. Investigating the pH of a water sample
Background information
Monitoring the physico-chemical properties of water is of vital importance.
Normal maximum permissible limit of pH for our life and health is 6.58.5.
Abnormal levels of pH and their consequences are given below:
pH 3 to 5 is too acidic for most organisms to survive, when the pH of water falls
below 4.5 most of the fishes die, leaving only a small number of acid-tolerant
insects such as water boatman and whirligig. These insects (beetles) can survive
and multiply even at pH 3.5. Similarly, pH>8.5 is too basic for most organisms
to survive.
Materials needed
Universal indicator test paper (broad range, narrow range PH 211)
Water sample
2. Investigating the biochemical (also called biological oxygen
demand [BOD]) of a water sample as pollution indicator.
Background information
A dissolved oxygen (DO) test measures the current status of oxygen in a water
body. This is a useful starting point.
However, DO content can vary considerably from day-to-day as affected by
many factors like temperature, wind velocity, eutrophication, pollution, etc.
The unpolluted water is characteristically rich in DO and low in BOD.
Higher the BOD, lower would be DO. Conversely, the polluted water has
high values of BOD. Water for drinking should have a BOD less than 1. Typical
BOD value for raw sewage run from 200400 mg of oxygen/ litre. The maximum


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96
LABORATORY MANUAL: BIOLOGY
Concentration Indicator organism Site 1 Site 2
Organisms in order of
tendency to disappear
as degree of pollution
increases
Red sledge worm (Tubifex worm)
Larvae of midge (Chironomus)
Blood Worm
Leech (Hirudinea)
Water l ouse, water skaters
(Asellus)
Fresh water shrimp (Gammarus)
Water boatman
Diving beetle (Dytiscus)
Caddisfly larva (Ochrotricha)
Damselfly larva (Lestes)
Stonyfly nymph (Isoperla)
Mayfly nymph (Stenonema)
Snail (Lymnaea)
Clams (Corbicula)
Fungi
Bacteria (anaerobic)
Utricularia
Chara
Water fern (Salvinia)
Water velvet (Azolla)
Water meal (Wolffia)
Lesser duckweed (Lemna minor)
Greater duckweed (Spirodela)
Diatom
A
R
X
X
X
X
X
X
X
X
X
X
X
none
none
X
X
X
X
X
X
X
X
X
X
A
A
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
3. Population density of plants
(i) Identify any 5 weeds from your locality.
(ii) Collect information about them from various sources. Focus on their
economic importance especially their medicinal importance (collect
samples for herbarium preparation).
permissible limit of BOD followed by Central Pollution Control Board (India)
for national water quality monitoring purposes is less than or equal to 3mg/ L.
For example, in a study the prevalence of some organisms were done at
two different sites in a water body. The result can be tabulated on the basis of
following facts:
A =Abundant; R =Rare; X =Very few


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97
INVESTIGATORY PROJ ECT WORK
(iii) Study their distribution in different localities by quadrat method.
(iv) Register their data and draw comparisons of their distribution by
histograms.
(v) Try to analyse the differences in distribution and density.
(vi) Correlate their presence with their habitat/ adaptability.
4. To make an inventory of local tree, shrub and herb
Informations can be listed in following categories:
(i) Avenue trees
(ii) Wind Breakers
(iii) Road dividers
(iv) Sound barriers
(v) Medicinal and other uses
5. Agrochemicals and their effects
The project may be carried out in a survey mode with a questionnaire prepared
with the help of the teacher to cover the following aspects.
(i) List of pesticides used, amount used/ hectare or acre, periodicity of spray,
name of the crop plant grown, recommended dose and the dose employed,
known effects on pest, whether the chemical pesticide is biodegradable
or not, alternate ecofriendly biocides.
(ii) List of fertilisers used, cost incurred/ acre/ year, recommended dosage/
time of use and the dosage used, known effects on fertility of soil, any
decrease in crop productivity, use of ecofriendly biofertilisers (VAM fungi,
leaf mold, green manure, dung, etc.).
6. Ecological role of some animals observed in a local area
Record the various plant species growing in the area under studytrees,
shrubs, annual/ perennial herbs, etc.
Note the season when flower/ seed is formed.
Note the various types of insects, birds, reptiles, amphibians, mammals, etc.,
and record their role as a/ an
(i) Herbivore
(ii) Pollinator
(iii) Agent in seed dispersal
(iv) Prey


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98
LABORATORY MANUAL: BIOLOGY
(v) Predator
(vi) Vectors for transmission of diseases
(vii) Any other
07. Study the effect of a local industry on environment
(i) Select an industry of your choice.
(ii) Note the source of energy used, product formed, raw materials used
(locally available or imported) mode of transport used to move the final
product.
(iii) Possible types of pollutants released by the industry (air/ water/
soil).
(iv) Measures undertaken by the management to comply with the standard
set by Central Pollution Control Board (CPCB), PCBs, etc.
(v) Awareness about ISO 2000.
(vi) Impact assessment carried out or not.
08. Study of the effect of chemicals and pollutants on the Mitotic
Index of the mitotically dividing onion root tip cells
This study may include
(i) Growing of onion root tip cells in the solution of pollutant/ chemical
and also in normal water as control.
(ii) Preparation and observation of slide for the study of mitotic index both
in experimental and control set-ups.
(iii) Analysis of the effect of pollutant/ chemical by comparing the data of
mitotic index between experimental and control variable.
09. Study of the genetic markers in the human population
In this investigation a few selected inherited traits can be investigated in the
family members of a small population in the locality. The compilation and
analysis of data will provide an about prevalence of trait in the said
population.
10. Inventory of weeds in aquatic bodies/agricultural fields
11. Inventory of birds in your locality, their ecological role as
scavangers, pollinators, etc.


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99
INVESTIGATORY PROJ ECT WORK
12. Impact of local industry on the environment and the remedial
measures taken by the industry
The guidelines and a brief outline of a few projects have been given with a
purpose to design and perform such investigation. Students and teachers can
think and design investigatory projects based on almost all concepts about
which experimental protocol have been given in the manual. However, a small
list of suggestive projects are also given below. Please note that these are only
suggestive and it is expected that students and teachers will take up many
more types of investigatory projects depending on the specificity of their area,
need and problems.


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Aim: To study the effect of pH on seed germination
Principle: pH is one of the most important factors that controls the composition of flora and
fauna in different terrestrial and aquatic ecosystems. pH of soils is essentially controlled by the
amount and type of various minerals and also by the quality and quantity of humus (dead,
decaying organic matter) present in it.
Seed germination is controlled by pH of the germinating medium. Seeds of different species
prefer a specific range of pH for maximum germination. pH not only controls the germination
of seeds but also growth and development, reproduction and various other metabolic activities,
of the plant.
Objectives: After completing the project, the students will be able to
1. Plan out an experiment and understand the use of appropriate chemicals, apparatus and
equipment, and learn the preparation of solutions.
2. Understand research methodology.
3. Generate, analyse and interpret the data and draw conclusions.
4. Conceive and choose other different themes related to pH and plant growth.
Materials required:
1. 125 seeds each of sunflower, mustard, green moong, alfa alfa, fenugreek and barley (selection
of seeds of different species may be made as per their availability)
2. Phosphate buffers
3. Distilled water
4. Petridishes of 7.5 cm diameter (15 pairs)
5. Blotting papers cut into circular discs to the size of petridishes
Procedure
(i) Prepare a range of pH buffers using Na
2
HPO
4
and KH
2
PO
4
.
(ii) Wash the seeds with water and blot them dry.
(iii) Select an appropriate place in the laboratory where there is sufficient
light. Arrange petridishes in three horizontal lines, with 5 dishes in each
line. Arrange petridishes horizontally in three rows A, B and C with seven
dishes in each row.
Investigatory Project 1


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101
INVESTIGATORY PROJECT 1
No and % of seed germination Total % of seed
germination
pH 1 2 3 4 5 6 7 A B C
4
5
6
7
8
(iv) Place one blotting paper disc in each petridish.
(v) Wet the blotting papers with small quantities of buffer solution of 4.0
pH for petridish No. 1, 5.0 pH for petridish No. 2 and so on till all the
blotting papers in petridishes No. 15 in row 'A' are wet with appropriate
buffer solution.
(vi) Repeat the process for rows B and C also.
(vii) Spread selected seeds in each of the five petridishes in such a way that
each petridish contains 25 seeds in row 'A'.
(viii) Repeat the process for rows B and C using two other types of seeds.
(ix) Cover the petridishes and record your observation after every 24 hours
for 7 days. Tabulate your results as given.
Observation
Observe the emergence of radicle as an indicator of germination and record in
the table. Calculate the percentage of germination every day.
Use the data of the table for graphic presentation.
Inferences and conclusion
The inferences and conclusion may be drawn on the basis of observations and the points given
below
Find out at what pH range seeds of different species had maximum % of germination.
Find out at what pH seeds failed to germinate or showed minimum % of germination.
Is there any general pattern of seed germination with regard to pH ranges?
What are the common features exhibited by the various types of seeds under varied pH
ranges? For example, did all the type of seeds show maximum germination in acidic range or
alkaline range or did pH preference varied between acidic and alkaline ranges?
Did you observe any relationship among time period, seed germination and pH range?
Table: Percentage of ................ seed germination


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Aim: Quantitative analysis of phytoplankton in a water body
Principle: The species composition and the density of the phytoplanktons determine the
productivity status of a water body. Phytoplanktons are the principal producers in a water
body. Based on the density of phytoplanktons, the water bodies are classified into non-
productive or oligotrophic and highly productive or eutrophic. Oligotrophic water bodies
support only a few species, whereas eutrophic water bodies support large number of species.
Further, the species composition of the phytoplanktons indicates the status of health of the
water body. Through phytoplankton assays, limnologists make an estimate of degree of pollution
in the water body. High density of cyanophycean algae, diatoms, volvocales, etc., are the
indicators of high degree of pollution. It should be noted that density and the species composition
of phytoplanktons exhibit diurnal, seasonal and annual fluctuations. It therefore becomes
important to monitor water bodies at regular intervals for drawing specific conclusions related
to their ecology.
It is in this context, the procedure for phytoplankton analysis on qualitative (species
composition) and quantitative estimation (density/ unit area) is suggested for students who
want to enter into fascinating realms of aquatic ecology.
Objectives: After completing the project, the students will be able to
1. Plan out an experiment.
2. Identify and quantify phytoplanktonic forms present in an aquatic ecosystem.
3. Interpret the data and draw conclusions.
4. Recognise the indicator species of pollution.
Requirements: Plankton net with plankton bucket, graduated plastic bucket (15 L), slides,
cover slips, compound microscope, watch glasses, dropper, and 5% F.A.A (Formaldehyde
Acetic acid, Alcohol)
Investigatory Project 2
Procedure
(i) Plankton net resembles the butterfly net in several aspects. Plankton
net, however, is prepared from bolting silk cloth which is readily available
at shops dealing in scientific equipments and chemicals. Procure about
one metre of bolting silk cloth of 40 mesh size and stich out of it a 40 cm


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103
INVESTIGATORY PROJECT 2
long cone with a diameter of about 20 cm at the mouth and a diameter
of 34 cm at the other end (both the sides open). Fasten to the mouth of
the cone a circular iron ring (with handle) of about 20 cm diameter with
the help of twine thread. Fasten a small steel bucket (plankton bucket)
or glass specimen tube of 50 ml capacity at the lower end.
(ii) Visit a nearby pond, pool or river bank carrying along with you the
plankton net fitted with the plankton bucket, graduated plastic bucket
and 510 ml of F.A.A.
(iii) Since this is a group activity, ask your friend to hold the plankton net
firmly a few centimetres above the water surface.
(iv) Immerse and fill the plastic bucket with water completely upto 15litre
graduated mark and filter the water through the plankton net. Repeat
the process several times (say10 times).
(v) At the end calculate the quantity of water in litres (X) filtered by
multiplying the amount of water in one bucket and number of buckets
of water filtered.
(vii) During this process of filtering, the planktons are collected in plankton
bucket. Only the water free of planktons escapes through the mesh of
the net.
Splash a few buckets of water against the net from outside taking care that
no water enters into the cone from the mouth. This will wash all the planktons
sticking against the inside wall of the net into the plankton bucket.
Detach the plankton bucket from the net and add a few drops (12 ml) of
5% F.A.A. to the plankton concentrate. Transfer the concentrate collected into
a suitable specimen tube and cork it. Note the volume of the concentrate (Y).
In the laboratory
1. With the help of 1 ml pipette, draw 1 ml of concentrate and transfer it
dropwise into the watch glass. Count the total number of drops that make
1 ml of concentrate (A).
2. Transfer one drop of plankton concentrate from the watch glass on a clean
slide. Cover it with square shaped cover slip. (For convenience divide the
area of the cover slip into parts with the help of lines drawn by Indian ink).
Observation
Observe the slide under microscope and count the number of total organisms
(B) by moving the slide from one corner of the cover slip to another horizontally
as well as vertically till the entire sample under the cover slip is completed.
With the help of following calculations find out the total number of different
organisms per litre of water.


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104
LABORATORY MANUAL: BIOLOGY
Unit cells/ L =
1000(ABY)
Unit/L =
x
Where A =number of drops in 1ml concentrate
B =number of organisms counted in 1 drop of concentrate
X =total amount of water filtered
Y =total volume of concentrate after filtration
Note: Another alternate method is the use of haemocytometer to calculate the
density of organisms under the guidance of teacher.
Inferences and conclusion
Find out the density and composition of organisms in different water samples
(polluted/ non-polluted).
Note the common organisms in both the water samples and those specific
to each sample.


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Notes


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Notes


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