PREFACE

In the ACs conference held in July, 2010 at KVS (HQ), New Delhi, issue of Study Material for Board classes was discussed at length and finally decided to provide it to students. Various Regional Offices were asked to prepare the study material in different subjects while the task of its correction and moderation was assigned to various ZIETs of KVS. KVS, ZIET, Chandigarh received study material in the subjects of Physics, Chemistry, and Biology &Maths for XII, Maths and Science &Tech. for X class, from various Regional Offices. The study material was got reviewed and suitably modified by organising workshops of experienced and competent subject teachers with the co-operation and guidance of AC,KVS,RO,CHD. Corrected study material was sent to all regional offices for providing it to students and also uploaded on the Website WWW.zietchandigarh.org. Subject teachers, both at the preparation and moderation levels have done a remarkable job by preparing a comprehensive study material of multiple utility .It has been carefully designed and prepared so as to promote better learning and encourage creativity in students through their increased self efforts for solving assignments of different difficulty level. But the teachers and the students must bear in mind that the purpose of the study material is in no way to replace the text-book, but to make it a complete set by supplementing it with this study material so that it may provide requisite and adequate material for use in different ways. The study material can be effectively used in the following ways: Practice material to supplement questions given in the textbook. Material for Study Camps: The purpose of conducting study camps is to inculcate study habits amongst students under active supervision of the teachers. These camps can beorganised within the normal school hours and days. Day wise target will be ascertained and given to the students and reviewed by the concerned subject teacher. If the target is not achieved by any student, it will be added to the next days target. Master Cards: The teachers can help students prepare master cards by taking the important questions/topics/points/concepts /reactions/terms etc from this study material for the quick revision for the examination. Crash Revision Courses: The material can also be used for preparing handouts for conducting Crash Revision Courses under the supervised guidance of the teachers just before or in the gaps between papers during examination.

Effectiveness of the study material will ultimately depend upon its regular and judicious use for the above listed purposes both by teachers and students. While attempting the source material, it would be quite useful to mark every time a question done successfully with a tick out ( ) and a question not done successfully with a dot ( ). It can be later used as a source of feedback for error analysis and for effective subsequent re visions/remedial work etc. I am sure that this well prepared study material if used sincerely and judiciously will surely bring cheers to all sections of students. I, also, take this opportunity to extend my most sincere gratitude to our Honble, Commissioner KVS (HQ), New Delhi, and other higher authorities of KVS for providing this opportunity for making some useful contribution to the study material. I also extend my thanks to all the Assistant Commissioners of various Regions for their in-valuable contribution in preparation of the Study Material in various subjects. Above all, sincere and dedicated efforts of the subject teachers in preparation of this study material deserve full appreciation . Teachers observations, suggestions and critical analysis for further improvement of the study material mailed to kvszietchd @g mail.com, will be highly appreciated. With best wishes to all users of this STUDY MATERIAL.

(HAR GOPAL) Director KVS ZIET Chd

STUDY MATERIAL FOR CLASS XII SESSION 2010-2011 BIOLOGY

K.V. OCF, SEC-29-B, CHANDIGARH.

How to use this study material

This study material is a supplement material to the NCERT textbook. It is neither a guide nor a refresher. The teachers can prepare the master card by taking the important topics/points/concepts /terms etc from this study material for the quick revision for the exam.

The material can also be used during the study camp by taking the important questions from the study material as mentioned in the level 1, 2& 3 assignments depending on the level of the student.

The material can also be used during the crash course by doing the revision of those topics by the teachers depending upon the topics given in starting or at the end of the chapter. Systematic revision of the different topics according to their level of difficulty & importance.

Details of the concept to be mastered by every child of class XII with concepts and exercises of NCERT Book. Symbols used * Important Questions * * Very Important Questions * * * Very-very Important Questions
S.NO 1 CHAPTER Reproduction In Organisms Concepts Types of reproduction A) Asexual reproduction B) Sexual reproduction, phases and events in sexual reproduction Degree of imp. Ref. NCERT text book.: page nos NCERT text book xii fig . 1.2(a) (b) fig. 1.3, 1.4 page 5-8 NCERT book p 15 - 19 ex q 2,6,9,13,15,18 Common errors fail to differentiate asexual reproductive structureszoospores, conidium, gemules etc. differentiation in monoecious & diocious 2 Sexual Reproduction In Flowering Plants 1. Pre fertilization: structures and events (i) stamens microsporangium & pollengrain microsporogenesis (ii) pistil megasporangium ( ovule) embryosacmegasporogenesis 2) pollination (i) autogamy , xenogamy, geitnogamy (ii) agents of pollination NCERT book fig 2.2. 2.3 , 2.5 p 21 23 NCERT book fig 2.7(d) 2.8 p 24 27 NCERT book p 27 28 NCERT book p 31 33 NCERT book p 34 fig 2.12. no. of cells in mature pollengrains no. of cells & nuclei in embryo sac , role of synergids self incompatibility triple fusion

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(iii) out breeding devices (iv) pollen pistil interaction 3. Double fertilization 4. Post fertilization : structures & events endosperm , embryo, seed 5. Apomixis - polyembryony 3 Human Reproduction 1 male reproductive system (i) diagram & description (ii) parts of male reproductive system (structure) (iii) functions of parts of system (iv) accessory ducts (v) accessory glands 2. Female reproductive system (i) diagram & description (ii) parts of female reproductive system (structure) (iii) functions of parts of system (iv) accessory ducts (v) uterus & its layers (vi) mammary glands

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(c,d,e) fig 2.13, 2.14, 2.15 p 35 NCERT p 38 NCERT NCERT P 43 , FIG 3.1 (B) NCERT P 43-44 NCERT P 43-44

free nuclear & cellular endosperm, embryo of monocot fail to differentiate apomixes , parthenocarpy

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Exact Location & Function Of Leydig Cells & Sertoli Cells

** * ** * ** * ** * *** *** *** Fig 3.6, page no 48 page no 48 NCERT P 44- 46 , FIG 3.3 (B) -DONCERT P 44-46 NCERT P 44 NCERT P 46 NCERT P 47 NCERT P 47 FIG 3.2 & 3.5 , 3.8 (a) P 49 Page no 47

3 gametogenesis (i) spermatogenesis & diagram (ii) stages of spermatogenesis with names of cells & no of chromosomes (iii) structure of sperm (diagram) (iv) functons of each part of sperm

Exact Stage Where Meiosis I & Ii Occurs During Gametogenesis As Well As The Ploidy Of Cells At Each Stage Of

& organelles (v) composition of semen 4 oogenesis i)structure and description ii) development of follicles iii) stages with names of cells and no. of chromosomes with events iv) significance of polar bodies 5 menstrual cycle (i) menarche and menopause (ii) phases of menstrual cycle with diagram (iii) role of hormones in cycle 6 fertilization and implantation (i) structure of ovum (ii) cleavage- formation of morula and blastula (iii )implantation- meaning, stage and site (iv) sex determination in humans (v) three germ layers 7 pregnancy and embryonic development (i) placenta as endocrine gland (ii) embryo and extra-embryonic layers 8 parturition (i) meaning (ii) foetal ejection reflex (iii) Role of hormones 9 lactation

Gametogenesis ** *** ** *** *** page no 48 Fig 3.7 ,Fig 3.8(b) Page no 48-49 Page no48-49 Page no48-49 Page no 49, 51 Fig 3.9 Co-relation of levels of pituitary hormones and events during menstrual cycle Difficulty in relating different stages of oogenesis with different life stages.

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Fig 3.1, Page no 51 Fig 3.11 Page no 52 Page 53 Page 52 Page 54

Labelling of mature graafian follicle

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Page 53 Fig 3.12 Page 53 Page no 54

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Hormones involved at the time of parturition

Reproductive Health

Meaning, colostrum and its importance 1. Reproductive health (i) Problems & Strategies 2. Methods of birth control 3. Infertility Corrective treatments 4. Sexually transmitted diseases 2 methods of birth control (i) natural methods (ii) barrier methods (iii) IUDs (iv)oral contraceptives (v) injections and implants (vi)surgical methods 4. Menstrual (i) Menarche and menopause (ii) Phases of Menstrual cycle with diagram (iii) Role of hormones in cycle 5. Fertilisation & implantation (i) structure of ouvum (ii) cleavage formation of morula, blastula (iii) implantation meaning, stage, site (iv) sex determination in humans (v) three germs layers 6. Pregnancy & embryonic development

Page no 54

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Page 57-58 Page 59-61 Page 64 Page 63

Amniocentesis Specific site for transplantation of embryo in GIFT and ZIFT

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49,51 fig 3.9, page 49-51

Fig 3.1, page 51 Fig 3.11, page 52 Page 53 Page 52 Page 54 Page 53

(i) placenta (ii) embryo & embryonic layers 7. Parturition (i) Meaning, role of hormones 8. Lactation (i) Meaning, colostrum and its importance 1. Mendels laws of inheritance (i) Reasons for choosing garden pea (ii) Seven contrasting traits of pea plant (iii) Symbols and terms used in mendels experiment (iv) Steps involved in mendels experiments (v) Monohybrid cross (vi) Test cross (vii) Incomplete dominance (viii) Codominance and multiple alleles (ix) Dihybrid cross 2. Chromosomal theory of inheritance 3. Linkage and recombination 4. Sex determination in animals 5. Mutations 6. Genetics disorder (i) Pedigree analysis (ii) Mendelian disorders (iii) Chromosomal disorder

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Fig 3.12

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Page 54

Hormones secreted by placenta

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Page 54

Principles of inheritance and variations

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Page 11/7 of Pradeeps Textbook Fig 5.1 page 7071 Page 71-73

** * *** *** *** *** *** ** ** *** ** ** *** *** Page 70-71 Page 71-75 Fig 5.5 74-75 Fig 5.6 page Table 5.3 page 77-78 Fig 5.7 page 79 Table 5.3 fig 5.8-5.9 Page 83-84 Fig 5.12 page 85-86 Page 87 Page 87-88 fig 5.13, 5.14 Page 89-90 Page 90-91 Heterogametey in sex determination Use of symbols for autosomal and sex linked disorders

Gamete formation in dihybrid cross

Molecular basis of inheritance

1. Dna (i) structure and salient features (ii) packaging of dna helix 2. Search for genetic material (i) transforming principle (ii) hershey and chase experiment (iii) properties of genetic material 3. Replication experimental proof 4. Transcription (i) transcription unit (ii) type of rna and process of transcription 5. Genetic code (ii)Mutations and Genetic Code (iii) TRNA-The adapter Molecule 6. Translation 7. Regulation of Gene Expression (i) Levels of Regulation (ii) The lac operon 8. Human Genome Project

** Page no. 96-98 ** Fig 6.4 page 99 *** *** ** *** Page 100-101 Fig 6.5 Page 102 Page 103 Differentiation between transformation and transaction Histone and non histone chromosomal protein Polarity of two stands

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Fig 6.7, 6.8 page 105-107 Leading and lagging strand direction Polycistronic, monoistronic, capping, taling Frame Shift and Point mutation Charging of tRNA Expresses Sequence Tags Sequence Annotation BAC/YAC Satellite DNA,VNTR

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Fig 6.9, page 107-108 109-111 page 112 Page 113 Fig. 6.12 Page-114 Fig. 6.13 Page 115 Page 115 Fig. 6.14,Page 116,117 Page118-120 Fi. 6.16 Page 121-122

9. DNA Finger printing

7.

Evolution

1. Origin of Life 2. Evidences of Evolution 3. Adaptive Radiation 4. Biological Evolution 5. Mechanism of Evolution 6. Hardy-Weinburg Principle 7. A Brief Account of Evolution 8. Origin and Evolution of Man

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Fig 7.1 Fig. 7.3 Page 130-132 Fig. 7.5,7.6,7.7 Page 133 Page 134 Page 135 Fig. 7.8 Page 136-137 Fig. 7.9,7.10 Fig. 7.11 Page 140 Fig. 8.1 Page 146-149

* 8. Human Health & Diseases 1.Common Diseases in human 2. Immunity (i) Innate immunity (ii) Aquired immunity (iii) Active and Passive immunity (iv) Vaccination and immunity (v) Allergy (vi) Autoimmunity (vii) Immune System of Body 3. AIDS 4.CANCER 5. Drug and alcohol abuse (i) Adolescence and drugs ** ** ***

Branching Desent and Natural Selection Darwinism versus devries- -Saltation Hardy-Weinburg Equlibrium,Founder Effect

Page 150-154 NCERT Fig. 8.4

Specific Role of histamines and cerotonins Mucosal associated lymphoid Tissue CMALT

* ** * **** *** **** Fig 8.6 Text 156 Text Page 156-158 Text Page 158-163

Contact Inhibition

9.

Strategies for Enhancement in food Production

1. Animal husbandry (i) Management of farm and farm Animal (ii) Animal Breeding (iii) Bee Keeping (iv) Fisheries 2. Plant breeding (i) Method (ii) For disease Resistance (iii) For Pest Resistance (iv) For Improved food quality 3. Single cell Protein 4. Tissue Culture 1. Role of Microbes in:(i) House Hold (ii) Industrial Product (iii) Sewage Treatment (iv) Production of Bio Gas (v) As Biocontrol Agent (vi) As Biofertlizers

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Page 170-176 Text Portion Page176 Text Portion Page 177 Text Portion

10.

Microbes in Human Welfare

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Text Page 181 Page 182,183 Ex. Question 12 Page 184-185 Ex. Question 7,8,11 Page 185 Page 186-187 Page 188

11.

Biotechnology principles and processes

1. Principles of biotechnology (i) techniques used in modern biotechnology (ii) advantages of sexual reproducton over a sexual reproduction (iii) genetic engineering includes reconibinant dna, genecloning a gene transfer (iv) meaning and use of plasmid restriction enzymen (v) basic steps for gmo 2. Tools of Recombinant DNA Technology 3. Cloning Vectors 4. Processes of Recombinant DNA Technology Steps *** *** ***

NCERT text book xii fig . 1.2(a) (b) fig. 1.3, 1.4 page 5-8

Fail to differentiate asexual reproductive structureszoospores,

NCERT book p 15 - 19 ex q 2,6,9,13,15,18

conidium, gemules etc.

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Differentiation in monoecious & diocious

Fig. 11.1-11.2-11.3 Page-195- 198 Fig. 11.4 Page-198-200 Fig. 11.6,11.7 Page 201-205

12.

Biotechnology and its applications

1. Applications of Biotechnology in agriculture (i) Advantages of GMO (ii)Bt Cotton (iii) RNA interference 2. Applications of Biotechnology in Medicines (i) Genetically engineered insulin. (ii) Gene Therapy-ADA (iii) Molecular Diagnosis of diseases. 3. Transgenic animals 4. Ethical issues , Biopiracy

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Page 207-208 Differentiation of Page 208 Page 208-209 Page 209-210 Cry and cry nRNA silencing , nematode Meloidegyne Page-210-211 Page 211 incognitia Steps in production of insulin Page-212 Role of Biotechnology in Page-213 Page-214 molecular diagnosis.

13.

Organisms and Populations

1. Organisms and its Environment (i) Major Abiotic Factors ** (ii) Responses to Abiotic Factors ** (iii) Adaptations *** 2. Populations:*** (i) (ii) (iii) (iv) Population Attributes *** Population Growth * Life History variation ** Population interactions Table 13.1 Page 232-238 Page 231-232 Fig. 13.5 Page 228-231 Fig. 13.4 Page 226-228 Page 225-226 Fig. 13.3 Page 223-225 Page -221-223 NCERT

Eurythermal & stenothermal Conformers, Regulators Distinction between Expanding, stable, declining population. Distinction between Exponential and Logistic growth curve. Distinction between commensalisms and Amaensalism.

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Ecosystem

1. Structure and function 2. Productivity * 3. Decomposition ** 4. Energy Flow ** 5. Ecological Pyramids 6. Ecological succession * 7. Nutrient cycling

Page 242 Ex Q. 9 Fig. 14.1 Page 243-244 Ex. Q10 Page 245,247 Ex. Q. 11 Fig. 14.4 Page 248 Fig. 14.5 Page 250-251 Fig. 14.6 Page 253-255 Ex. Q. 12,13 GPP,NPP

15.

Biodiversity and Conservation

1. Patterns of Biodiversity 2. Importance of species diversity to ecosystem. 3. Loss of Biodiversity 4. Conservation of Biodiversity

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Fig. 15.1 Text page 259 Ex. Q 3 Page 263 Page 264-265 Ex. Q. 5 Page 265-267 Ex. Q. 7

Graphical representation , species area relationships Crypreservation

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Environmental issues

1. Air Pollution and its control (i) Case study of Delhi

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Fig. 16.1 Page 270-273 Page 272-273

Advantages of CNG over Petrol or diesel. Norms of Air

2. Water pollution and its control (i) Domestic Sewage & Industrial effluents (ii) (iii) (iv) (v) Algal Bloom BOD Eutrophication Biomagnification *** *** *** *** Page 275 Page- 275 Page-276 Fig. 16.5 Page-276 ** Page-274 Fig.16.2,16.3

Pollution. Types of impurities & their nature in domestic sewage. Effect of Sewage discharge on characteristies of river

3. Solid waste (i) Case study of remedy for plastic waste. (ii) Electronic waste *** *** * ** Page-279 Page-279 Page-279 Page-280 Page-280

Concentraion of toxic substances at various trophic levels

4. Ago chemicals & their effects (i) Case study of organic farming 5. Radioactive wastes

6.Greenhouse effect and Global Warming (i) Green house gases & their relative contribution to total global warming 7. Ozone depletion in Stratosphere 8. Deforestation:- Case study of conservation. *** ** ***

Page-280-282 Fig. 16.6,16.7 Types of e-wastes & the metals extracted.

Page-282-283 Page-284-285

Role of UV-B radiations

Contents
S No Chapter 1 2 3 4 5 6 7 8 9 10 11 12 Reproduction in animal Sexual reproduction in flowering plants Human reproduction Reproductive health Principles of inheritance and variation Molecular basis of inheritance Evolution Human health and diseases Strategies for enhancement in food production Microbes in human welfare Biotechnology :principles and processes Biotechnology and its application Pages 0-11 12-17 18-36 37-46 47-63 64-96 97-105 106-124 125-130 131-142 143-167 168-172

13 14 15 16 17 18 19 20 21

Organisms and populations Ecosystem Biodiversity and conservation Environmental issues Wordlist Concept Mapping Word List Hots Assignments Assignments of different difficulty levels Assignments for self evaluation

173-183 184-192 193-195 196-208 208-209 210-214 215-246

CONTENTS
Sl No 1 2 3 4 5 6 7 8 9 10 11 12 13 Chapter Reproduction in animal Sexual reproduction in flowering plants Human reproduction Reproductive health Principles of inheritance and variation Molecular basis of inheritance Evolution Human health and diseases Strategies for enhancement in food production Microbes in human welfare Biotechnology:principles and processes Biotechnology and its application Organisms and populations Pages 0-11 12-17 18-36 37-46 47-63 64-96 97-105 106-124 125-130 131-142 143-167 168-172 173-183

14 15 16 17 18 19

Ecosystem Biodiversity and conservation Environmental issues Wordlist Concept Mapping Word List Hots Assignments

184-192 193-195 196-208 208-209 210-214 215-246

Chapter 1: - REPRODUCTION IN ORGANISMS

REPRODUCTION IN ORGANISMS Life Span Period from birth till natural death. Every organism live only for a certain period of time. Example: Elephant: 60-90 years Fruit fly: 4-5 weeks Reproduction Producing young-ones of their kind, generation after generation. Types of reproduction: Asexual reproduction :single parent capable of producing offsring. Sexual reproduction : two parents are invovled in producing offspring. Modes of asexual reproduction Binary fission: parent body divides into two halves, genetically identical to parent. Amoeba: It is simple or irregular.

Paramoecium: Transverse binary fission. Multiple fission: parent body divides into many daughter organisms : Plasmodium. Budding: daughter organisms grow from small buds arising in parent body. Exogenous budding: out side the body eg. Hydra, Yeast.

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Budding in Yeast Endogenous budding : inside the body eg. Gemmule in sponge. Conidia: non-motile, exogenous spores in chains eg. Fungi. Zoospores: microscopic motile structures eg. Algae. In plants : term vegetative reproduction frequently used instead of asexual reproduction, units of vegetative propagation called vegetative propagules.

Adventitious buds in Bryophyllum

Sexual reproduction All organisms show remarkable similarity Vast difference in their reproductive structure. Similar pattern or phases in their life cycles.

Juvenile phase: The phase of growth before reproductive maturity. Reproductive phase: Reproductive maturity. Senescent phase: Phase between reproductive maturity and death.

The main events of sexual cycle are: i. Prefertilisation events: a.Gametogenesis : The process of formation of male and female gametes by meiosis(cell-division).

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Homogamete(Isogamete): - gametes similar eg. Algae Heterogamete(an-isogamete): - morphologically dissimilar gamete ,male gamete (antherzoid or sperm) ,female gamete (egg or ovum) eg. Human. Sexuality in organisms : In plants Bisexual term is used for Homothallic and Monoecious plants Both male and female reproductive structures in same plant eg. Higher plants, cucurbits and coconut. Unisexual term is used for Heterothallic and Dioecious plants Male and female reproductive-structure on different plants., Flowering plants male flowerstaminate flower and female flowerpistillate flower eg. papaya and date-palm. Animals Bisexual term is used for Hermaphrodite animals-eg. Earth-worm, Tapeworm, Leech, Sponge. Unisexual animals have male & female sexes in separate individuals-e.g. insects, frogs, humanbeings Cell division during gamete formation: Hapliod-parent (n) produces haploid gametes (n) by mitotic division, eg. Monera,fungi, algae and bryophytes. Dipliod parent (2n) produces haploid gametes(n) by meiosis division (possess only one set of chromosomes)and such specialized parent cell is called meiocyte or gamete mother cell. ExampleName of organism Human Housefly Ophioglossum (fern) in Meiocyte(2n) 46 12 1260 2 6 gamete (n) 23 6 630

Potato

48

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b) Gamete transfer:- to facilitate fusion. Male gametes mostly motile and female non-motile,exception few fungi and in algae both gametes are motile in some cases Water medium for gamete transfer- in lower plants. Large number of male gametes produced to compensate loss Higher plants, pollen-grains are transferred by pollination. Fertilization: Fusion of male and female gametes diploid zygote. Parthenogenesis.-development into new organism without fertilisation eg. Rotifers, honey-bees, some lizard, bird(turkey). Fertilization Two types- external and internal . External fertilisation- outside the body of organism in external- medium (water) eg. majority of algae, fishes, amphibians. Advantageshow great synchrony between the sexes 1.release of large number of gametes into surrounding medium 2.large number of offsprings produced. Disadvantage- offsprings are extremely vulnerable to predators, natural disasters. Internal fertilisation- fusion occurs inside female body eg. majority of plants and animals. Egg non-motile and formed inside female body. Male gamete motile, produced in large numbers to reach egg and fuse with it. In seed plants, non- motile male gamete carried to female gamete by pollen-tube.

Post -fertilisation events- formation of zygote. a. zygote. One celled , diploid, vital link between two generations. External fertilization zygote formed in external medium water eg. Frog, Internal fertilization zygote formed inside the body eg. Humanbeings. Development of zygote depends on type of life cycle and environment. Some develo thick wall ( prevent damage and desiccation) & undergo period of rest eg. algae, fungi. In haplontic life cyclezygote (2n) divides by meiosis to form haploid (n)spores. Diplontic life-cycle- zygote (2n) divides mitotically, develops into embryo(2n). Oviparous animals lay eggs out-side the female body.Eggs can be fertilized/ unfertilized. Fertilized eggs covered which hard calcareous shell, laid in safe place in the enviroment. Unfertilised eggs laid in water. Example- fishes, frogs, reptiles, birds 2 7

Viviparous animals bear and rear the embryo inside female body, give birth to youngones.Advantage- proper embryonic care, protection, survival chances of young-ones greater. Example- cows, whales, humanbeings Embryogenesis: development of embryo from zygote by cell division (mitosis) and cell differentiation. Cell- division increases the number of cells in the developing embryo formation of

Cell differentiation - groups of cells undergo certain modifications for the different kinds of tissues and organs.

In flowering plants- zygote formed inside ovule Changes occur in flowering plants: sepals, petals, stamen- wither and fall-off. Zygote develops into embryo. Ovule forms the seed (germinates to produce new plants). Ovary transformed into fruit which develops protective thick wall-pericarp.

Parthenogenesis: Female gamete develops into new organism. Seedless fruits can be formed by parthenogenesis Clone: A group of individuals of the same species that are morphologically and genetically similar to each other & their parents

Very short answer type(1 mark) What is meiocyte? Specialized cells in diploid organism, i.e., gamete mother cell which undergo meiosis. Name the kind of reproduction in bees by which drones are produced? Parthenogenesis. What is special in flowering bamboo? Bamboo species flower only once in their life-times generally after 50-100 years. 2 8

What is meant by homothallic? The term homothallic refers to bisexual or hermaphrodite condition.

Why are the date palms referred to as dioecious ? In date-palms, the male and female flowers are present in different plants.

If the diploid number of chromosomes in an angiosperm plant is 28, what number would you expect in the endosperm and embryo of that plant? Endosperm: 48 Embryo: 28

Give the scientific terms for the following

a. Morphologically and genetically similar individual derived through asexual reproduction. b. Cyclical changes shown by seasonal breeders. a. Clone b. Oetrous cycle Short answer type (2 marks) Name the structure which gets transformed into seeds at maturity. ovule Name any one animal in which self-fertilization occurs. Taenia (tapeworm) What is the site of origin of new plantlets in the followings ?

a. Potato tuber, b. rhizome of ginger, c. leaves of bryophyllum, d. stem cutting of sugar cane a.Buds called eyes b.Adventitious buds at Nodes c.Advevtitious buds from the notches at margins of leaves d.A portion of stem with buds at nodes 7

What do the fallowing parts in a flower form after fertilization?

a. Zygote, b. ovule, c. ovary-wall, d. Petal a.Embryo b.Seed c.Pericarp d.Wither and fall off Draw a diagram of the cross section of a simple fruit and label two important parts-

Show diagrammatically only reproduction in yeast.

Short type answer (3 marks) Q Write the mode of asexual reproduction in the following organisms:

Penicillium, Spongilla, Paramoecium, Yeast, Chlamydomonas, Amoeba. Ans:Penicillium: Conida.

Spongilla: Internal budding (gemmules). Paramoecium: Binary fission (transverse). Yeast: Budding (external). Amoeba: Binary fission (simple). Chlamydomonas: Zoospore. Q Differentiate between: External Fertilization / Internal fertilization, Zoospore / zygote, Gametogenesis / Embryogenesis.

External fertilization 1. Occurs outside the body of females.

Internal fertilization Inside the body female

2.

Large number of gametes are released into the surrounding medium.

Number of gametes produced is less.

Ex- fishes, frogs

Ex- lizard, humans

Gametogenesis Haploid gametes formed

Embryogenesis Multicelluar embryo from cd unicellur from zygote Involves cell division, cell growth, celldifferentiation

Two types: spermatogenesis (formation of male gamete) and oogenesis (formation of female gamete Meiosis occurs.

Mitosis occur

Zoospores 9

Zygote

Formed inside zoosporangium Flagellated and motile Result of asexual reproduction Haploid or dipliod

Fusion of two gametes Usually nonflagetted and non motile or motile Result of sexual reproduction Dipliod

Q. What are the three major phases in the life cycle of organism? Define each phase Ans. The three major phases are: i. Juvenile phase. ii. Reproductive phase. iii. Senescent phase. Juvenile phase: The Phase of growth before reproductive maturity. Reproductive phase: Attainment of reproductive maturity. Some major hormonal changes mark this phase. Senescent phase: The phase between reproductive maturity and death. Q. What is external fertilization? Name two animals having external fertilization. Why are more gametes produced by such animals? Fertilization occurs outside the body of organism Frog and bony-fish Large no of gametes to enhance produced the chances of fertilization

Long answer type question(5 marks) Discuss the similarities in pattern of sexual reproduction. A) Pre-fertilization: Prior to the fusion of gametes after attainment of maturity. This includes two main events: i. Gametogenesis: Formation of two types of gametes male and female which involves spermatogenesis in males and oogenesis in females. ii. Gamete transfer: It is to facilitate fusion. Medium by water, pollination, copulation. B) Fertilization: Fusion or syngamy. 10

C) Post-fertilization: Single celled zygote. Embryogenesis: Process of Development of embryo from zygote. Zygote undergoescell-division and cell-differentiation. By cell-division- it increases the number of cell cell-differentiation- group of cells undergo certain modifications to form specialized tissues and organs to form an organism. HOTS: FIVE MARKS QUESTIONS: 1. Study the following diagram and answer the questions given below:

i. What does S and P denote? ii. Where do they develop from? iii. What is the term given to the point of attachment of the ovules in the fruit? iv. What is the ploidy of embryo and the tissues in the ovary? v. What is the function of fruit apart from storage of materials? 2. Fertilization leads to the formation of embryo. i. Give the technical term for the development of embryo. ii. What are the events that occur during embryo development? iii. The development of zygote depends on two factors. What are they? iv. How will you categorise animals based on the development of zygote inside or outside the female body? v. How does zygote in fungi and algae overcome dessication?

Chapter 2 SEXUAL REPRODUCTION IN FLOWERING PLANTS

FLOWERS Site of sexual reproduction. Male & fema le reproductive organs are borne on flower s.

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a PARTS OF A FOWER: 4 WHORLS - CALYX (sepals), COROLLA(petals), ANDROECIUM( Male reproductive organ), GYNOECIUM( Female reproductive organ). Male reproductive organ Androecium consists of stamens Stamen consists of anther , filament & connective( when anther is bilobed) Anther is bilobed & has 4 microsporangia

L.S. OF A FLOWER WITH DIFFERENT PARTS

MICROSPOROGENESIS: Microspore mother cell (2n)

Meiosis Microspore (n)

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Mitosis Pollen grains (n)

Pollen grains have two outer walls; i) Exine ii) Intine Exine is made of sporopollenin (HARDEST NATURAL SUBSTANCE) Mature pollen grains have two cells big vegetative cell & small generative cell. Generative cell forms two male gametes by mitotic division. Pollen grains shed in 2-celled/3celled stage GYNOECIUM / CARPEL (THE FEMALE REPRODUUCTIVE ORGAN)

-Each Carpel consists of ovary, style & stigma.

- Ovules are attached to ovary by placenta.

. Structure of an Anatrpous ovule

The funicle stalk of ovule Hilum, a region where funicle is attached Integuments cover embryo sac. Micropyle a pore for entry of pollen tube & to imbibe water

Parts of gynoecium 13

Structure of anatropous ovule

MEGASPOROGENESIS Megaspore mother cell (2n) Meiosis 4 Megaspores (n) (3 megaspores degenerate, 1 remains functional)

Funtional Megaspore (n) (Divides 3 times by mitosis)

8 Nucleated Embryo Sac formed

DEVELOPMENT OF 8 NUCLEATED EMBRYO SAC

14

3 cells group at micropylar end -the egg cell(n) & 2 synergids(n)

3 cells at chalazal end called antipodals(n) 2 polar nuclei at center(n each )

POLLINATION transfer of pollen from anther to stigma. Agents of pollination air, water, insects.bat,bird,man.

Double fertilization

syngamy 1st male gamete fertilizes egg. nd 2 male gamete fertilizers fusion product of polar nuclei. Pollen grains germinate on stigma & pollen tube grows through style. Pollen tube reaches micropyle & releases two male gametes into embryo-sac. Fertilisation is the process of fusion of male& female gametes (n+n) to form a diploid(2n) zygote SYNGAMY: Fusion of one male gamete(n) with egg (n) fusion Fusion of two polar nuclei(n+n=2n) Second fusion Zygote(2n) produced First

The other male gamete(n) fuses with the fusion product of two polar nuclei(2n) fusion

Third

DOUBLE FERTILISATION: i ) Fusion of male gamete with egg First fertilization, ii )Fusion of fusion product of polar nuclei with male gamete Second fertilisation POST FERTILISATION CHANGES: 15

Stages of development of the embryo after fertilation : STAGES OF EMBRYO DEVELOPMENT AFTER FERTILISATION 1. Zygote divides by mitosis into suspensor & embryo cells 2. Suspensor cell forms a globular basal cell which remains embedded in the endosperm & a multicellular suspensor bearing the embryo 3. Globular embryo becomes heart-shaped & then mature embryo with radicle, plumule & cotyledons Primary endosperm nucleus divides repeatedly to form endosperm, food for the embryo. Mature ovary becomes fruit. Mature ovule beomes seed. True Fruit develops only from the ovary, e.g. mango, tomato False Fruit develops from parts of the flower other than the ovary,e.g. apple, pe Seeds are of two types:

16

Dicot seed

Monocot seed

OUTBREEDING DEVICES: Continued self-pollination result in breeding depression. Flowering plants have developed many devices to discourage self-pollination & encourage cross-pollination such as Bearing unisexual flowers Anthers & stigmas mature at different times Anthers & stigmas are placed at different positions Self-incompatibility where pollen grains of a flower donot germinate on the stigma of the same flower ARTIFICIAL HYBRIDISATION: Type of cross-pollination performed by man for crop improvement. Achieved by- i ) Emasculation i.e.removal of anthers from the flower bud of a bisexual flower before the anther dehisces using a pair of forceps and ii )Bagging i.e.covering the emasculated flowers with a bag of suitable size to protect them from contamination with unwanted pollen If flower unisexual, emasculation is not needed. Flower bud bagged & when the stigma becomes receptive, pollination is done using desired pollen & the flower is rebagged

Emasculation

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Chapter 3: Human Reproduction

The Male reproductive system
1. Penis a. urination b. sexual intercourse 1. Corpus cavernosum- spongy tissue that fills with blood to make penis erect 2. Glans- the head, end of penis 3. Foreskin i. Covers glans, ii. May be removed surgically in an operation (circumcision) a. Located behind penis b. Contains two testes c. Temperature sensitive (Sperm must be made in cooler conditions i.e, 2-3 C lower than body temperature)

2. Scrotum

3. Testes a. Sperm is produced by the seminiferous tubules due to FSH b. Testosterone is produced by Leydig cells due to LH 1. Causes the development of the male sex organs at ~8 weeks after conception. 2. Responsible for facial, armpit, and pubic hair, bone growth and muscular development. c. Testes are formed in the abdomen before birth. Descend through the "inguinal canal" during fetal or post-natal life. Sometimes may take months/years to reach right place. Possible site for hernia.

4. Epididymis: 5. Vas deferens: 6. Prostate gland: 7. Seminal Vesicles: 8. Cowper's gland:

Stores sperm til mature Tube from the epididymis to urethra sperm cells stored here. Provides an alkaline fluid that can protect sperm from harsh vaginal acids. Produce food purpose for sperm. Produces clear lubricating fluid

Hormonal control of male reproductive system 1. Pituitary gland at the base of the brain produces:
a. FSH (follicle stimulating hormone) 1. b. FSH regulates spermatogenesis.

LH (luteinizing hormone) 1. LH prompts Leydig cells to produce testosterone, (male hormone.) 18

Hormonal control of female reproductive system 1. Pituitary gland (at the base of the brain produces):
a. FSH (follicle stimulating hormone) i. ii. b. Responsible for maturation of follicles and respective ova. Stimulates estrogen production.

LH (luteinizing hormone) i. Ovulation- release of ovum from ovary usually on 14th day of menstrual cycle. Ovulation generally occurs every 28 days. Causes the corpus luteum to produces progesterone and estrogen. Progesterone hormone of pregnancy.

ii.

c.

Estrogen: the main female hormone produced in Graffian follicles of ovary -

1. increases size ( growth and development) of oviducts, uterus, and vagina. 2. responsible for secondary sexual characteristics: 3. Fat in breasts and development of duct system. 4. Broadening of the pelvis. 5. Soft and smooth skin. 6. Fat in buttocks and thighs. 7. Pubic hair.. d. Oxytocin causes uterine contraction during Parturition. e. Progesterone produced by corpus luteum. (ruptured graffian follicle after ovulation) The placenta continues to produce progesterone during the pregnancy. 1. 2. 3. pregnancy hormone calms muscular contractions of the uterus. It (along with estrogen) thickens the lining of the uterus.

f. Hormones from Placenta: hCG: human chorionic gonadotrophin hPL : human placental lactogen

Female Reproductive System.

2. Ovaries produce:
a. Ova: I) ii) each ovary contains immature ova (eggs) in follicles. Females are born with lifetime supply of eggs. (250,000-400,000 in each ovary) 19

iii)

Ovaries release ovum -. Almost all ova degenerate between birth and puberty. iv) Approx. 400 eggs will be ovulated over woman's life. v) An egg is the largest human cell. vi) Ovaries are located lower abdomen. One on either side. vii) Egg is viable for only 24-48 hours after ovulation.

2. Fallopian tubes
a. b. c. d. e. Two thin tubes attached to the upper sides of uterus Tubes terminate near the ovaries but not attached "Fimbriae" finger-like structures on the end of each tube Tubes conduct egg to uterus by use of small hairs called "cilia" Fertilization of ovum takes place in the ampullary-isthmic junction of the fallopian tubes. Pear-shaped located in lower abdomen Muscles (myometrium) stretch to allow baby to develop. Oxytocin starts labor contractions. Lining of uterus (endometrium) thickens with blood-rich tissue due to progesterone Endometrium supports embryo/fetus during growth Placenta (the after birth) attaches to endometrium. Interface between baby and mother. If not pregnant, lining breaks down and is discharged from body through vagina. This is menstruation (period) Cervix connects uterus to vagina. Like a door that opens during ovulation. Cervical mucous closes the door at all other times.

3. Uterus
a. b. c. d. e. f. g.

4. Vagina
a. Birth canal b. Menstrual blood leaves the body c. Organ of intercourse d. Very muscular stretches to allow a baby to grow ? e. Vaginal opening remains partly closed by thin membrane of tissue called hymenf. Hymen may be stretched or torn during any physical activity

6. Cervix
a. b. c. d. e. Located at inner end of vagina Opening of uterus into vagina Mucous prevents bacteria and viruses from entering uterus Lets sperm into uterus after ovulation Baby passes through during vaginal birth

7. Labia a. 2 layers of skin, which fold over the opening to vagina and urethra i. Inner labia (labia minora) ii. Outer labia (labia majora) site of pubic hair growth

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8. Clitoris
a. Small organ, 5 to 10 millimeters long b. Located at junction of inner labia near front of body c. Contains erectile tissue d. Sexually sensitive 9. Mons pubis Cushion like fatty tissue covered by skin and pubic hair

Difference between

1) Spermatogenesis and oogenesis

Spermatogenesis Produces male gametes (sperm) occurs in the seminiferous tubules (in testes)

Oogenesis Produces female gametes (oocytes) occurs in the ovaries

involves meiosis occurs throughout life after puberty

involves meiosis occurs after puberty until menopause

may produce 400,000,000 per day

humans normally produce one oocyte during each ovarian cycle Primary otocyte divide unequally to form one large secondary oocyte and a small polar body An oogonium produces one functional ovum and 3 non functional polar bodies

Primary spermatocyte divide equally to form two similar secondary spermatocytes One spermatogonium produces 4 functional spermatozoa

Role of amniotic fluid 1 Prevents desiccation 2 Shock absorber

21

Menstrual Cycle
Menstruation Repair of the endometrium Ovulation Thickening of the endometrium Breaking down

Follicular Phase FSH/Estrogen

Due to LH

Luteal Phase LH/Progesterone

2)Follicular phase (Proliferative phase ) and Luteal phase (secretory Phase)

Proliferative phase
Also called Follicular phase Stage of repair and proliferation It extends from the end of menstruation to ovulation LH and FSH increases Estrogrn level inceases Estrogen is secreted by Graffian follicle

Secretory Phase
Also called Luteal phase Prepares endometrium for implantation extends after ovulation to menstruation

LH is high (LH surge) Progesterone level increases Progesterone secreted by corpus luteum

22

The menstrual can be represented as 23

Notes on FSH and LH FSH and LH from the pituitary: Hormone FSH Controls LH Controls
.The follicle stages

FUNCTION / PRODUCTION In Females Eggs + Estrogen Ovulation + Corpus Luteum In Males Spermatogenesis Testosterone

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From primordial follicle to tertiary follicle

Primordial follicle
the surviving primary oocytes , at birth, are surrounded by thin, single layers cells of so-called follicular epithelial cells.

Primary follicle
Primordial follicles with iso or prismatic follicuilar follicles the follicular epithelium surrounding the oocyte becomes iso- to highly prismatic

A B 1 2

Primordial follicle Primary follicle Oocyte Follicular epithelium

1 2 3 4

Oocyte Pellucid zone Stratum granulosum Theca folliculi cells

Secondary follicle
Follicles with multiple rows of follicular epithelium stratum granulosum. Pellucid zone:- Zone between the oocyte and follicular epithelium.

Tertiary follicle
Follcile with well-developed net of capillaries in the theca interna. Antrum a fluid filled cavity. Theca layers of cells organized into external Theca externa and interna linterna.

Tertiary follicle

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1 2 3 4 5 6 7 8

Oocyte Pellucid zone Stratum granulosum Theca interna Theca externa Antral follicle Cumulus oophorus (Granulosa cells, together with the oocyte) Basal lamina between theca and stratum granulosum

Schematic stage From fertilization to embryo

Sperm + secondary oocyte

Fertilized ovum (zygote) 2-cell stage 4-cell stage 8-cell stage Morula Blastula Early Gastrula Late Gastrula

Cleavage-

cells division of embryo similar to mitotic division 26

MorphogenesisDifferentiationGrowth-

stage distinct shapes start to appear formation of distinct cells and tissues

cells get bigger and so does embryo

Conception to Birth
The following shows some of the many stages of human development:

Zygote

The single cell that results from fertilization of an ovum by a sperm.

Morula

=
*The morula (little mulberry)I solid ball (16 64 cells. Morula results of mitotic (cleavage)divisions.

Blastocyst

The blastocyst is a liquid-filled ball of cells. This stage of development occurs around 5 8 days after conception. Implantation in the endometrium occurs at this stage.

27

Embryo

The human is considered an embryo from implantation until about 8 weeks after conception.

Fetus =
8 weeks after conception until birth.

The fate of three germ layer

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Ectoderm Nervous system

Mesoderm
Skeleton

Endoderm Digestive tract

Epidermis of skin

Muscles

Respiratory system

Circulatory system

Liver, pancreas

Gonads

Bladder

Mnemonics
Tubules in male reproductive system 29

SEVEN UP Seminiferous tubules Epididymis Vas deferens Ejaculatory duct (Nothing) Urethra Penis Menstrual Cycle FOL(d) M(a)PS Ovarian cycle: Follicular phase Ovulatory phase, Luteal phase D Menstrual cycle: Menstrual flow, A Proliferative phase, Secretory phase (The ovarian cycle controls the menstrual cycle. The cycles begin (day 0) when menstrual flow starts. At day 14, ovulation take place. The luteal and secretory phases begin and last until day 28,after which the cycles begin again.)

TERMS TO REMEMBER Acrosome- the part of a sperm cell that contains enzymes (This enables a sperm cell to penetrate an egg.) Afterbirth-placenta and fetal membrane expelled from the uterus after the birth of a baby Amniotic sac-fluid-filled membrane or sac that surrounds the developing embryo while in the uterus; ". (Protects the baby from hard shocks and keeps it at a constant temperature. ) Birth-the process of being born; the baby moves from the uterus into the outside world. Parturition Blastocyst = blastula = early stage of an embryo surrounded by a liquid-filled sphere whose wall is composed of a single layer of cells A+ this stage (about eight days after fertilization) implantation in the wall of the uterus occurs. Cervix- the lower part, or neck, of the uterus. (Opening to the uterus.)
30

Clitoris-small, sensitive organ in front of the vagina Coitus-synonym for sexual intercourse Conception-fertilization of an egg cell by a sperm cell Corpus Luteum - Crater-like structure formed after ovulation produces progesterone and estrogen. Old RUPTURED GRAFFIAN follicle. It means yellow body . Cowper's glands - 2 glands that secrete an oily liquid, which cleans and lubricates the urethra of the male Egg cell-a female sex cell (female gamete or ovum or secondary oocyte ) Ejaculation-the discharge of semen from the penis Embryo-the unborn child developing in the uterus between the second and eighth weeks of life Endometrium - the innermost lining for the uterus, Site for blastocyst to implants and develop. Epididymis- tightly coiled tube at the back of each testis holds newly produced sperm (each epididymis is like a nursery where sperm mature and learn to swim.) Erection-the condition of the penis when it fills with blood and becomes firm, enlarged, and erect. Fallopian tube -tube one from each ovary leading to the uterus. Carry the egg from the ovary to the uterus. Fertilization occurs here. (Also known as the oviducts.) Fimbria - (plural fimbriae) means "fringe." a fringe of tissue near the ovary leading to the fallopian tube Fertile- able to conceive a child Fertilization-Union of a sperm and an egg. Conception. Fertilized egg- Egg after a sperm has united with it. Zygote Fetus- Unborn child developing in the uterus after the first eight weeks of life Follicle - Found in ovaries. Each holds and nourishes the egg until ovulation. It is like a nest. Will become corpus luteum after ovulation. Foreskin A sheath of skin that surrounds the penis. Follicle Stimulating Hormone - hormone secreted from pituitary gland in both men and women. In women, FSH promotes the development of eggs and estrogen. In men, FSH promotes the development of sperm cells. Fraternal twins-babies that develop from two eggs, each fertilized by a sperm cell; may or may not be of the same sex Gamete Sex cells. Sperm cells and egg cells are gametes. Genitals-Reproductive or sex organ. especially the external organs Germ Cell- An egg or sperm cell. A gamete. In humans, a germ cell contains 23 chromosomes Hormones-chemical substances produced by the endocrine glands Act in other parts of the body and affect maturation, growth, and behavior; LH, FSH, GH, Testosterone, Estrogen, Progesterone are all hormones. Hymen-a thin tissue or membrane that may partially cover the opening of the vagina Identical twins-babies that develop from a single fertilized egg that separates into two halves. Identical twins always of the same sex Labia-two folds of skin surrounding the entrance of the vagina Labor-the muscular contractions that expel a baby from the uterus during childbirth Leydig Cells when prompted by LH, Leydig cells create testosterone. LH - Luteinizing Hormone - Secreted from the pituitary gland and causes ovulation and the formation of the corpus Luteum in women. In men, LH causes the Leydig cells to produce testosterone.
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Menstrual cycle -The periodic building up and sloughing off of the lining of the uterus approximately every twenty-eight days Menstruation- The periodic discharge of blood and waste material (unfertilized secondary oocyte / ova and the degenerating endometrium lining) from the uterus Miscarriage-expulsion from the uterus of a fetus before it is developed sufficiently to live Spontaneous abortion Myometrium muscles of the uterus that stretch to accommodate the growth of the baby. These muscles contract during labor and push baby out Orgasm-the peak of sexual excitement when the male ejaculates semen. Ovaryfemale sex glands. Produces egg cells, estrogen and progesterone. OvulationDischarge or release, of an egg cell / secondary oocyte from the ovary Ovumscientific name for an egg cell / secondary oocyte Oxytocin - Hormone, released from the pituitary gland , stimulates contraction of the myometrium of the uterus during labor and facilitates ejection of milk from the breast during nursing. Penis-the male sex organ through which sperm cells leave the body. Also discharges urine Placenta- network of blood vessels and other tissues by which the unborn child is attached to the wall of the uterus. Grows out of the endometrium The umbilical cord is attached to it. It is the interface between mother and developing fetus. Pregnancy-the process in a woman from conception to birth Pregnant-the condition of a woman with an embryo or fetus in her uterus Progesterone pregnancy hormone. First produced by the corpus luteum and then by the placenta. *Progesterone increases lining of endometrium. *maintains pregnancy * helps in development of mammary glands. Prostate gland- Organ that surrounds the upper end of the male urethra and produces part of the fluid that mixes with the sperm to form semen. Prostate fluid is alkaline and helps to protect sperm from vaginal acids. Scrotum-Pouch of loose skin containing the testes. This houses and air-conditions the testicles by moving and sweating. Semen-Mixture of sperm and fluids that appears at ejaculation. Semen is comprised of sperm, fructose, prostate fluid and oil from the Cowpers gland. Seminal vesicles-small saclike organs opening into each vas deferens near the upper end of the urethra. Produce part of the fluid that mixes with the sperm to form semen. Provide food (fructose) for the sperm. Seminiferous tubules - Tubes found in the testes that produce sperm Sexual intercourse- The entry of the penis into the vagina and the subsequent release of semen coitus Sperm- The male sex cell (male gamete or spermatozoon), which contains 23 chromosomes in humans. Spermatozoon- Scientific name for sperm Spontaneous abortion-- Synonym for miscarriage Testes- Male sex glands. Produce sperm cells and testosterone. Testicles-synonym for testes Testosterone Male hormone that regulates the development of the penis, muscles, body hair, etc
32

Umbilical cord-the ropelike structure connecting the embryo or fetus to the placenta within the uterus. Urethra-the tube through which urine is expelled from the bladder in both males and females and through which semen leaves the male body Uterus = womb - the hollow pear-shaped organ in which a baby develops before it is born; Vagina-the passage from the uterus to the outside of the body. - accepts the penis during intercourse. -Birth canal. -Menstrual fluids leave through it Vas deferens-The tube extending from each epididymis to the urethra in males Womb- Synonym for uterus Yolk sac- Structure that develops for the nutrition of the embryo during early embryonic life and then ceases to function Zygote - Cell formed by the union of two gametes, [ fertilized ovum before cleavage.]
********************************* Practice material Chapter 3 HUMAN REPRODUCTION

1 mark questions Where does fertilization takes place in human female? Where and at what stage implantation takes place? What harm is caused if the testis fail to descend into the scrotal sac? Name the layer of cells forming the outer layer of blastula? What is semen? Why progesterone is called pregnancy hormone Name the hormones secreted by a. leydigs cell b. graffian follicles c. Corpus luteum d. Placenta 2 marks questions 8. Differentiate between a. Spermiogenesis and spermatogenesis b. Follicular phase (proliferative phase) and Luteal phase (secretory phase) c. menarch and menopause d. Spermatogenesis and oogenesis e. morulla and blastulla 1. 2. 3. 4. 5. 6. 7.

9. 10. 11. 12.

How is ovulation different from menstruation? How does the ovum ensure that only one sperm fertilizes it? Name the accessory reproductive glands in human male along with its secretion and functions What is colostrum? What is its significance?

33

Placenta acts as an endocrine gland why it is said so ? What are its other function? Where does oogenesis occurs in human?. Describe the stages of the process Where does Spermatogenesis occurs in human?. Describe the stages of the process What is menstruation? Mention the specific action of FSH,LH,estrogen and progesterone in menstrual cycle? HOTS 13. 14. 15. 16. 17. Suppose the acrosome of mammalian spermatozoa does not function normally, how would it affect fertilization? Give reasons. 18. An ovum allows the entry of only one sperm at a time. Why? 19. Failure of fertilisation leads to menstruation. Explain

20. Explain why there is no menstrual cycle before puberty , after menopause and during pregnancy. 21. Fill in the boxes

22. Study the diagram and answer questions that follows:

34

a.What is the role of corpus luteum in menstrual cycle? b. Why is luteal phase otherwise called as secretory phase? c. How is estrogen related to menstrual cycle? d. How does FSH and LH regulate the menstrual cycle?

23 What is the fate of inner cell mass in the blastocyst? Mention their significance. 24 Label a,b,c in the following diagram. 3

35

Give the term / reason 5 a) Mechanism responsible for parturition. b) Role of oxytocin during expulsion of the baby out of uterus c) Why does zona pellucida layer block the entry of additional sperms? d) Sperm cannot reach ovum without seminal plasma. e) All copulations do not lead to fertilization and pregnancy Furnish the technical term for the following: 5 a) Cushion of fatty tissue covered by skin and pubic hair in female external genitalia. b) The finger like projections which collect ovum after ovulation c) The middle thick layer/wall of uterus d) Semen without sperm e) The finger-like projections appearing on the trophoblast after implantation.

36

Chapter-4:REPRODUCTIVE HEALTH
Reproductive Health: Human reproductive health and sexuality involve great many components and interrelationships. A total view of human reproductive health is basic to personal well-being as well as to interpersonal relationships. Every individual is a unique sexual being. Adolescents are vivrant, fragile and prone to experimentation and risk taking , as a result they are the most vulnerable population as far as delinquent behavior and attitude are concerned. Every decision has its own consequence. Any wrong decision can lead to disastrous consequence, which in turn can ruin ones life. Sexual adjustment is part of total personality adjustment. Self-esteem is the key to sexual maturity. Broad based community and institutional support for reproductive health is essential.

Adolescence Reproductive and Sexual Health (ARSH) Importance of ARSH reducing specific risky behaviours theories which explain what influences people's sexual choices and behaviour . clear, and continuously reinforced message about sexual behaviour and risk reduction . Provide accurate information about, the risks associated with sexual activity, about contraception and birth control, and about methods of avoiding or deferring intercourse Dealing with peer and other social pressures on young people; providing opportunities to practise communication, negotiation and assertion skills . Uses a variety of approaches to teaching and learning that involve and engage young people and help them to personalise the information. Uses approaches to teaching and learning which are appropriate to young people's age, experience and cultural background . Creating a protective and supportive environment. Avoiding manipulative relationships. Promoting and protecting the sexual and reproductive rights of adolescents and youth .

37

Methods of birth control Behavioural methods: Behavioural methods depend on a good knowledge of the menstrual cycle as well as adequate self control by the couple. o Coitus Interruptus: 'Interrupted sex". Penis is withdrawn from the vagina just before ejaculation. Advantage No drug,. No interference with normal body functions. Planned pregnancy. Disadvantage Dependent almost wholly on the man's self-control. Failure rate high 15 - 18%. Rhythm method or Safe Period: Requires knowledge of the female partner's menstrual cycle to identify the days on which sexual intercourse is possible without the risk of pregnancy. Read more ... o Avoiding vaginal Intercourse: Anal sex, oral sex or sex without penetrating the vagina. Sexual act occurs between the tightly held thighs of the woman has no risk of pregnancy. Barrier methods: Barrier placed between the penis and the vagina during intercourse so that the sperm cannot meet the ovum for fertilization. o Male Condoms: Male condoms consist of a sheath, usually made of latex that covers the erect penis during penetration of the vagina. Read more... o Female Condoms: Not used often. Made of polyurethrane. Consists of a loose sheath with two rings on either side. The smaller ring is pushed deep into the vagina , larger ring remains outside on the edge of the vagina. The female condom can be inserted about 8 hours prior to sexual intercourse and can be kept in for about another 12 hours after intercourse. Can be used more than once during this period. o Condoms protect against pregnancy as well as sexually transmitted diseases (STDs), including HIV/AIDS. o Diaphragm: Vaginal diaphragm small saucer shaped rubber sheath with a metal coil in its rim which is fitted across the mouth of the uterus (cervix). o Cervical Cap: Small dome-shaped rubber device fitted on the cervix. Uncomfortable to apply and is rarely used nowadays. o Vaginal Sponge: Small polyurethrane round device which needs to be placed inside the vagina before sexual intercourse. It releases spermicide wich makes sperm inactive .Be left in place for 8 hours after use and can be used more than once during this time. The sponge also acts as a barrier contraceptive to some extent since it swells up to fit across the cervix once it is inside the vagina. Hormonal Methods: In this method, drugs are used to either prevent ovulation or to prevent implantation of the embryo after fertilization. Combined oral contraceptives contain two hormones similar to the natural hormones in a womans body---an estrogen and a progestin.
o

38

Working of Birth Control Pill : Preventing ovulation. In normal menstrual cycle, the pituitary gland secretes the hormones FSH and LH to stimulate the ovary to release an egg ('ovulation"). When pills are taken, the level of estrogen in the blood increased. This sends a negative feedback message to the pituitary gland to stop secreting FSH and LH. The lowered levels of FSH and LH fails to stimulate the ovaries and ovulation is prevented. The progesterone in the pills make the cervical mucus hostile to the sperm. .It also work by causing changes that make the endometrium unreceptive to a fertilized ovum if ovulation and fertilization do take place
o

o o

Oral Contraceptive pills: Combined oral contraceptive pills or birth control pills contain two hormones - estrogen and progesterone. One to prevent ovulation. Second to disrupt the normal growth of the internal uterine lining (endometrium) so that the embryo cannot implant in it. Read more... Centchroman: Non-hormonal non steroidal contraceptive. Function: slow down growth rate of internal uterine lining and speed up the movement of the embryo so that implantation cannot occur. Read more... The Patch: (Ortho Evra). Thin band-aid like patch containing estrogen and progesterone which should be applied over the skin. It releases the hormones slowly into the skin through which they are absorbed. Depo-provera: Injecting a high dose of the hormone progesterone every three months. prevents ovulation. Disadvantage : irregular bleeding throughout the three months. Nuvaring: Thin silastic ring inserted into the vagina once every month. Releases the hormones estrogen and progesterone and prevents ovulation during the menstrual cycle. Subdermal Implants Norplant (a registered trademark of The Population Council for levonorgestrel subdermal implants) set of six small plastic capsules. The capsules are placed under the skin of a womans upper arm. Norplant capsules contain aprogestin, similar to natural hormone of a womans body makes. Hormone released very slowly from all six capsules. Capsules supply a steady, very low dose. Contain no estrogen. Capsules thicken cervical mucus making it difficult for sperm. Stops ovulation (release of eggs from ovaries) in about half of the menstrual cycles after first year of use. Emergency Oral Contraception After unprotected sex, emergency oral contraception can prevent pregnancy. Sometimes called postcoital or morning after contraception. Mainly stops ovulation (release of egg from ovary). Perhaps also works in other ways. The sooner emergency control contraceptives used, the better they prevent pregnancy. Up to 72 hours after unprotected sex, the woman should take 4 low-dose or 2 "standarddose" combined oral contraceptives, and then take another equal dose 12 hours later. Regular use of emergency contraceptives has serious health hazards. Vaginal Pessaries, Tablets, Creams or Foams : Contain spermicides which are toxic to the 39

sperm and should be inserted into the vagina just before coitus. Advantage : easy to apply, do not interfere with coitus , act as lubricants. Disadvantage : not very effective always. Intra-Uterine Contraceptive Devices (IUCD): IUCDs or IUDs contraceptive devices which are placed inside the uterus. Usually small, flexible plastic frame. Often has copper wire or copper sleeves on it. It is inserted into a womans vagina through her uterus. Almost all brands of IUDs have two strings, or threads, tied to them. The strings hang through the opening of the cervix into the vagina. A provider can remove the IUD by pulling gently on the strings with forceps. IUDs work chiefly by preventing sperm and egg from meeting. IUD makes it hard for sperm to move through the womans reproductive tract. Reduces ability of sperm to fertilize the egg. Could also prevent the egg from implanting itself in the wall of the uterus. IUCDs prevent pregnancy by making the endometrium unreceptive to the fertilized ovum. Stimulates the endometrium to release leukocytes (WBCs) and prostaglandins making it hostile to the sperm. Causes bizarre and irregular growth of the endometrium. Prevents implantation of a fertilized ovum. IUDs like Copper-Ts also come wrapped in copper. The copper is toxic to sperms and is a method of enhancing the contraceptive effect of the IUDs. The IUCDs can come in various shapes and sizes. Lippes Loop: The Lippes loop consists of a thin plastic (or polyethylene)wire bent in a series of S-shapes. Needs to be straightened when inserted into the uterus but resumes its shape once inside.

l Lippes loop Copper-Ts

Copper T: Cu-T common IUCD. T-shaped structure stays inside the uterus with the long arm of the T along the uterine cavity (endometrium), shorter arms transversely across the upper part of the endometrium. Copper-Ts contain a copper wire or copper bars of different strengths wrapped around the arms or the stem. A Copper T 250 has 250 sq mm of copper wire wrapped around it and is effective for 3 years. The most commonly used IUCD is the Copper T 380. 40

Mirena: Hormone based IUD releases a progesterone called levonorgestrel. Works by affecting ovulation, affecting the normal growth of the endometrium and by affecting the cervical mucus so that the movement of sperm obstructed. In United Kingdom, hormone based IUDs are known as Intra-uterine Systems (IUS). Surgical Methods: These are more or less permanent methods of contraception. o Tubal Ligation: Both female tubes tied off and usually cut during tubal ligation to prevent the sperm from reaching the ovum during intercourse. o Vasectomy: The two tubes in the males i.e. vas deferens which carry sperm from the testes to the penis are tied off and cutt. o Essure: Method in which small micro-inserts are placed at the mouth of the fallopian tubes Cause scarring and block them. Prevents sperm from reaching the ovum for fertilization. PREGNANCY P occurs when a sperm produced by a male, fuses with an ovum produced by a female. Every month a mature ovum released from either one of a womans two ovaries. Ovum round and about 100 microns in diameter, slightly smaller than the period at the end of this sentence. Largest cell in the human body and Only one visible to the naked eye. Ovum picked up by the fallopian tube on the same side. (There are two fallopian tubes on either side of the uterus near the ovaries. See image ). Tubes have long fingerlike projections called fimbria which are used like hands to pick up the ovum. The ovum then moves through the tube, propelled by long hairs growing from cells in the tubes. Like grass bending before the wind, hairs bend towards the uterus in waves, pushing the ovum slowly towards the uterus. Egg remains viable, that is, alive for about 72 hours, but is capable of being fertilized for only about 12 24 hours. If it remains unfertilized during this period, it disintegrates in the tube without leaving any trace. Its end products (mainly proteins) are absorbed into the bloodstream and excreted through the urine or stool.

41

Fertilization, Implantation and Pregnancy Sperm however is viable for a longer period, been found in the uterus 5 7 days after coitus. Capable of fertilizing an ovum for only 48 - 72 hours after being ejaculated. Time taken by the sperm to reach the tubes is between 6 12 hours, Sometimes as early as 1 hour. Intercourse has to take place within this narrow time frame (1-2 days before ovulation or immediately after ovulation), for a pregnancy to occur. At every intercourse a normal man deposits 2 5 mililitre of semen in the upper part of the vagina (see diagram). Each mililitre of semen contains about 50 200 million of sperms. Whisking their tails madly, the sperms swim rapidly upwards into the uterus and from there into the two tubes on either side at the rate of 3mm per hour. It takes an average of 10 hours for the sperm to reach the tubes. All the sperm deposited in the vagina cannot swim into the uterus. Some die off in the vagina, some get entangled in the cervical mucous and some manage to swim just into the cervix before dying. But it is believed that even these sperms help in causing pregnancy by changing the acidity (ph) of the vagina or by acting on the cervical mucous so that other sperms can penetrate it and reach the ovum. Only about 1 % of the total number of sperms deposited in the vagina make the journey. Thousands of sperms flood the uterus and both the tubes. Some tumble out of the opening of the tubes into the abdomen. Hundreds of sperm (estimated to be around 300) surround the ovum in the tube. They press against the membrane of the ovum attempting to penetrate it and fertilize the ovum. Finally one sperm succeeds. At once a chemical reaction is triggered off in the wall of the ovum, making it impenetrable to any other sperm. No other sperm can enter the ovum now.

42

The unsuccessful sperms slowly degenerate. The sperm the ovum fuses with it to form zygote. The zygote starts to divide as it is propelled towards the uterus dividing Zygote called embryo. In-vitro fertilization (IVF ovum and the sperm is allowed to fertilize in a laboratory dish (petri dish). Embryo usually transferred into the mothers uterus on 3rd day at the 4 8 celled stage. The zygote reaches the uterus usually on 6th to 9th day after ovulation, Morula a 16-celled. Separation between 2-celled to 16-celled stages (identical twins). Separation at later stages can lead to the potentially fatal condition of conjoint twins or Siamese twins. Morula sticks to the inner lining of the uterus ('endometrium'). Progesterone from ovaries prepares the endometrium to receive morula. Morula burrowed deep into endometrium by 9th - 12th day Can cause implantation bleeding. Development of embryo continues until 9th months of pregnancy (40 weeks or 280 days), to form fully formed baby. INFERTILITY AND ITS TREATMENT

Infertility:- Woman fails to conceive after one year of sexual life without contraception. According to (WHO) World Health Organisation, infertility about 10 % worldwide. Infertility treatment, azoospermia (complete absence of sperm). Intra-Uterine Insemination (IUI): Used when men has moderately low sperm count. Semen collected by masturbation, washed and centrifuged to increase the sperm density. Sample injected into the uterus. Procedure within 2 hours of collection of semen.
o o o

Insemination with Husband's Semen (AIH) Impenetrable cervical mucous or diseased deformed Impotence or premature ejaculation. Insemination with Donor Semen (AID) Semen from somebody other than husband.

InVitro Fertilisation (IVF)) is a process by which egg cells are fertilised by sperm (usually 100,000 sperm / ml) outside the womb, in vitro. Process hormonal controll ovulatory process, removing ova (eggs) letting sperm fertilise in a fluid medium. Fertilised egg (zygote) transferred to the patient's uterus. The first successful birth of a "test tube baby", Louise Brown, occurred in 1978. 43

In vitro, within the glass, In vivo procedure, tissue remains inside the living organism within which it is normally found. IVF, also calledtest tube babies, Zygote intrafallopian transfer (ZIFT) fallopian tubes prevents binding of sperm to egg. Egg cells removed from ovaries, and in fertilised blocked vitro . Resulting zygote is placed into the fallopian tube by laparoscopy. Gamete intrafallopian transfer (GIFT). ZIFT success rate 64.8% one cycle five weeks. Steps: Woman to take fertiolity medication stimulate egg production. Ovaries follicles mature woman infected with HCC (human chorionic gonadotropin. Eggs harvested 36 hours later, by transvaginal ovum retrieval. Fertilization in laboratory resultant early embryos or zygotes placed woman's fallopian tubes by laparoscope. Gamete intrafallopian transfer (GIFT) Eggs removed from woman's ovaries, Placed in one Fallopian tubes, along with the man's sperm. Technique, pioneered by endocrinologist Ricardo Asch, Fertilization inside the woman's body. [1] Four to six weeks to complete a cycle of GIFT. Harvested egg, mixed with the man's sperm, placed into the woman's Fallopian tubes using a laparoscope. Intracytoplasmic Sperm Injection (ICSI): Single sperm injected into centre of the egg, to achieve fertilization. Advantage used for men with very low sperm count In azoospermic, men sperm suctioned out of the vas deferens ( male tubes). Careful testicular biopsy and MESA - Microepididymal sperm aspiration. Prevention of Male Infertility : Undescended testes treated infancy. Infections by mumps and other viruses should be managed by medically. General Facts About STDs Sexually transmitted diseases STD, or STIs (sexually transmitted infections) are transferred from one person to another through sexual contact. 25 diseases can be transmitted through sexual activity. E.G. HIV, chlamydia, gonorrhea, syphilis, genital herpes, human papillomavirus, hepatitis B, trichomoniasis, and bacterial vaginosis. Risk prone: Adolescents. Pelvic inflammatory disease, cause infertility. STDs can be prevented by refraining from sexual activity, some contraceptive devices, such as condoms. Specific STDs: An Overview Human Papilloma Virus: Main cause of cervical cancer. Treatment other types of cancers of the female reproductive system. Reduces signs and symptoms, no cure. HPV vaccine developed. Herpes Virus: Symptoms Periodical blisters or sores on the genitals. Hepatitis: Hepatitis B (HBV) vaccine available. Other hepatitis infections through sexual contact include Hepatitis A and Hepatitis C. HIV/AIDS: One STD that many people are worried about getting is HIV. While new ways of treating this 44

infection can significantly prolong an infected person's life, for far too many people this infection eventually progresses to AIDS and, ulitmately, death. More than 40 million people worldwide are infected with the HIV virus; women account for 50% of those infected. Auto Immuno Deficiency synotrome, Caused by HIV. Syphilis: Syphilis can easily be treated and cured. Symptoms can progress and affect the nervous system and brain leading to dementia and death. Trichomoniasis:Common, curable STDs. Common Infections: Chlamydia and gonorrhea often infect a person at the same time. Can be cured Damage reproductive system if left untreated. Pubic Lice: Crabs are very similar to head lice. Symptom itching. Can be treated. Rare Infections: Granuloma inguinale and chancroid, Well known in North America, Lesser-talked about STDs nongonococcal urethritis, molluscum contagiosum

KNOW THE SYMPTOMS OF STDs

Swelling or tenderness in genital area. Blisters ,sores or bumps around the mouth or genitals. Fever,chills and aches. Unusual itching. Burning sensation when you pass urine or move your bowels. White,watery or yellow disharge from the penis.

Women face special risks

Usually have fewer symptoms than men, often none at all. Bleeding that is not part of their period. Pelvic or vaginal pain. Discharge from the vagina. Painful urination. Unusual rash, sore or growth in the genital area.

DONT LET STDs TO TAKE YOU BY SURPRISE Pelvic Inflammatory Disease Overview Pelvic inflammatory disease (PID) is infection of a woman's reproductive organs. Infection spreads upward from the cervix to uterus, Fallopian tubes, ovaries, and surrounding structures Symptoms Lower Abdominal pain or tenderness Back pain Abnormal uterine bleeding Unusual or heavy vaginal discharge Painful urination 45

Painful sexual intercourse Symptoms not related to the female reproductive organs include fever, nausea, and vomiting. PID symptoms may be worse at the end of a menstrual period and during the first several days following a period. Ectopic Pregnancy Overview Pregnancy that develops outside a woman's uterus (womb). Fertilized egg develops somewhere else in the abdomen especially fallopian tubes, outside of Utres, on ovaries or attached to bowels. Fetuses destroyed. Usually found in first 5 10 weeks of pregnancy. Complications Intra-abdominal hemorrhage (severe bleeding) rupture of fallopian tube.

Chapter 5 Genetics
Relationship between genes and chromosome of diploid organism and the terms used to describe them 46

Know the terms

Terms

Meaning Address/ location of a gene in a chromosome

Example T,A.b,d etc T and t OR A and a etc AA or aa Aa or Tt etc AA

Allele Homozygous Heterozygous Homozygous Dominant Homozygous recessive

Allelomorphs= alternative form of a gene Both alleles of a gene at a locus are similar Both alleles of a gene at a locus are dissimilar Both alleles of a gene at a locus are similar & dominant Both alleles of a gene at a locus are similar & recessive

aa

Mendel's first law ( Law of dominance )characters are controlled by discrete units called genes (allele) which occur in pair .in heterozygous condition only one gene can express it self which is dominant. (Can be explained by monohybrid cross) Mendel's second law ( Law of segregation): The two alleles received one from each parent segregate independently in gamete formation, so that each gamete receives one or the other with equal probability. (Can be explained by monohybrid cross) 47

Mendel's third law ( Law of recombination): Two characters determined by two unlinked genes are recombined at random in gametic formation, so that they segregate independently of each other, each according to the first law (note that recombination here is not used to mean crossing-over in meiosis). (Can be explained by dihybrid cross) This is what Mendel said (summary) : 1) Dominant alleles overpower recessive alleles. Dominant traits overpower recessive traits. 2) Rule of segregation (Separation): Gametes (sex cells) only receive one allele from the original gene. 3) Rule of Independent assortment: One trait will not determine the random selection of another.

In complete dominance: When one allele of a gene is not completely dominant over the other and the F1 hybrids are intermediate between two parents. The phenotypic and genotypic ratio is same.1:2:1 in F2 generation. E.g. Snapdragon or Antirrhinum majus Co dominance: Two alleles of a gene are equally expressive and dominant in a generation eg Human blood group ( Note : Human blood group is also an example for multiple allelisim i,e when a gene exists in more than two allelic form) Basic outline of Mendels cross 1. Pure breeding parents for a pair of contrasting character (allelic Pair) is taken e.g, Tall pure-bred pea plants (TT) & short pure-bred pea plants (tt)

2. Gamete formation (Meiosis)

3. Hybridization (crossing is done)

4. F1 generation - the product of the above cross (are called hybrids)

5. Selfing (allowed to self fertilize / self breeding )

6. Gamete formation (Meiosis)

7. F2 generation - the product of the above selfing 48

8. Analysis of result (Phenotype and Genotype) Tendency of genes of same chromosome to remain together Such genes are called linked genes. Linked genes present only parental types

Figure 1 Schematic diagram of Genetic Linkage and Recombination (A) Two homologous chromosomes: blue (paternal) and orange (maternal). Three genes with separate alleles and linkage " noted (A,a; B,b; C,c;). (B) Crossing over during meiosis. ( (C) Two alleles and their linked genes have switched locations via recombination. Four additional alleles and their associated (A,a; B,b;) have not switched and are considered linked. (D) Recombined haploid chromosomes segregate separately during meiosis as gametes before fertilization. (E) Sample recombination frequencies between genes demonstrating higher rates of recombination for genes further apart. 49

Cross Monohybrid Tt X Tt Dihybrid cross YyRr X YyRr Incomplete dominance Rr X Rr 1:2:1 3:1 9:3:3:1

Result of F2 generation 1:2:1 1:2:1:2:4:2:1:2:1

1:2:1

Co Dominance and multiple allelisim Blood group A B AB O Possible genotype IAIA OR IAi IBIB OR IBi IAIB ii

Crosses of blood group ( CO DOMINANCE) Blood group AXA Possible genotype IAIA X IAIA IAIA X IAi IAi X IAi BXB IBIB X IBIB IBIB X IBi IBi X IBi AB X AB OXO IAIB X IAIB ii X ii Possible phenotype A A A;O B B B; O AB: A; B O

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POSSIBLE BLOOD GROUP OF PROGENY WITH RESPECT TO THE BLOOD GROUP OF PARENTS

A AXA AXO AXB BXB BXO AB X A AB X B AB X O AB X AB OXO KEY + + + + + + + + = POSSIBLE

B + + + + + + + -

AB + + +

O + + + + + -

+ -

- = NOT POSSIBLE

Sex determination and sex chromosome Organism Human beings Birds Insects Male XY ZZ XO Female XX ZW XX

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Pedigree Analysis

Pedigree is a chart of graphic representation of record of inheritance of a trait through several generations in a family Symbols used

Four patterns of inheritance

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AUTOSOMAL DOMINANT

AUTOSOMAL RECESSIVE 1. Traits are controlled by recessive genes and appears only when homozygous 2. Both males and females are equally affected 3. Traits may skip generations 4. 3:1ratio between normal and affected. 5. Appearance of affected children from normal parents (heterozygous) 6. All children of affected parents are also affected. 7. e.g.- Albinism, sickle cell anaemia etc

1. Traits are controlled by dominant genes 2. Both males and females are equally affected 3. traits do not skip generations 4. e.g. polydactyly, tongue rolling ability etc

X-LINKED DOMINANT

X-LINKED RECESSIVE

1. All daughters of an affected male will be affected 2. there is no male to male transmission. 3. e.g very rare-Ritts disease

1. Traits is more common in males (since hemizygous) but rarely in females (since homozygous) 2. all daughters of a male who has the trait are heterozygous carriers 4. there is no male to male transmission. 5. affected males receive the trait from carrier mother who has affected father 6. e.g- haemophilia, colour blindness

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A 'typical' autosomal recessive pedigree

54

A 'typical' autosomal dominant pedigree

x-chromosome linked pedigree Now try to answer

1. Is it possible that this pedigree is for an autosomal dominant trait? 2. can two individuals that have an autosomal dominant trait have unaffected children?

3. Is it possible that this pedigree is for an autosomal dominant trait?

4. Is it possible that this pedigree is for an autosomal dominant trait?

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5. Is it possible that the pedigree above is for an autosomal recessive trait? 6. Assuming that the trait is recessive, write the genotype of each individual next to the symbol A = normal a = the trait (a genetic disease or abnormality)

7. Is it possible that the pedigree above is for an autosomal recessive trait?

8. Write the genotype of each individual next to the symbol

9. Is it possible that the pedigree above is for an autosomal recessive trait?

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10. Is it possible that the pedigree above is for an X-linked recessive trait? 11. Write the genotype next to the symbol for each person in the pedigree

12. Is it possible that the pedigree above is for an X-linked recessive trait?

13. Is it possible that the pedigree above is for an X-linked recessive trait?

14. Is it possible that the pedigree above is for an X-linked recessive trait?

15. Is it possible that the pedigree above is for an X-linked recessive trait?

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16. Is it possible that the pedigree above is for an X-linked recessive trait? Clues Affected Autosomal Dominant AA Aa Unaffected aa

Autosomal Recessive

aa

AA Aa

X- chromosome linked ecessive

X X X Y

XX XX XY

TERMINOLOGIES Allele = A factor or letter that makes up a gene. 2 alleles make up one gene. Alternative forms of a genetic locus; a single allele for each locus is inherited separately from each parent (e.g., at a locus for eye color the allele might result in blue or brown eyes). Alleles = "B" and "b" are different alleles. Autosomal genes = genes that are not found on the sex chromosomes. Autosomal chromosomes are ones that are not XX and XY. A chromosome not involved in sex determination. The diploid human genome consists of 46 chromosomes, 22 pairs of autosomes, and 1 pair of sex chromosomes (the X and Y chromosomes). Carrier = A person who has a defective gene and a dominant normal gene and therefore, is normal . (Nn) Centimorgan (cM): A unit of measure of recombination frequency. One centimorgan is equal to a 1% chance that a marker at one genetic locus will be separated from a marker at a second locus due to crossing over in a single generation. In human beings, 1 centimorgan is equivalent, on average, to 1 million base pairs Chromosomes = 46 are found in human cells. Genes are carried on chromosomes. 58

Clones: Group of cells derived from a single ancestor. Cystic Fibrosis = Autosomal recessive. Mucous in lungs... Death in the 20s. DNA = The main instructions that explain how to build the body. (Deoxyribose Nucleic Acid) DNA makes up alleles, genes and chromosomes. Dominance = This is one of Johann Gregor Mendels principles. In his studies with pea plants Mendel notices that pure tall plants bred to pure short plants resulted in tall hybrid plants. Tallness was dominant over shortness. Dominant = A allele that overpowers another is dominant. Down's Syndrome = Due to an extra chromosome. (21st pair). Gamete = Sperm or egg. Germ Cell. In humans, germ cell contains 23 chromosomes. Genetics: The study of the patterns of inheritance of specific traits Gene = Every trait is controlled by a gene. A human has 20,000 genes. Genes are controlled by 2 factors called alleles . Each allele comes from a parent. Genotype = Hereditary or Genetic constitution of an Organism. (Genes an organism possesses) Genome: All the genetic material in the chromosomes of a particular organism; its size is generally given as its total number of base pairs. Germ Cell- An egg or sperm cell. A gamete. In humans, a germ cell contains 23 chromosomes. Haploid= A single set of chromosomes (half the full set of genetic material), present in the egg and sperm cells of animals and in the egg and pollen cells of plants. Human beings have 23 chromosomes in their reproductive cells. Hemophilia = Sex-linked recessive. Males get it most often. Heterozygous = This means alleles of a gene are "different". Heterozygosity=The presence of different alleles at one or more loci on homologous chromosomes. Homozygous = When alleles of a gene are "the same" Homologous chromosomes: A pair of chromosomes containing the same linear gene sequences, each derived from one parent Huntington's Chorea = Autosomal Dominant. People die at 40 +... Jerky muscular motions Hybrid = Means alleles of a gene are "different" (Hh) See heterozygous.

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Independent Assortment: This is Johann Gregor Mendels 2nd principle. It states that alleles of one gene separate independently from alleles of another gene. In other words, eye color does not affect a persons ability to roll his or her tongue.

In vivo: Inside living Organism.

In vitro: Outside a living organism in artificial or laboratory conditions. Karyotype: A photomicrograph of an individuals chromosomes arranged in a standard format showing the number, size, and shape of each chromosome type; Linkage: The proximity of two or more genes on a chromosome; the closer together the genes are, the lower the probability that they will be separated during meiosis and hence the greater the probability that they will be inherited together. Linkage map: A map of the relative positions of genetic loci on a chromosome, determined on the basis of how often the loci are inherited together. Distance is measured in centimorgans (cM). Locus (pl. loci): The position on a chromosome of a gene or other chromosome marker; also, the DNA at that position. The use of locus is sometimes restricted to mean regions of DNA that are expressed.( See gene expression).

Meiosis = The kind of cell division that produces sperm and egg. Meiosis reduces the number of chromosomes to half. In humans, for instance, the nuclei of body cells contain 46 chromosomes. Due to meiosis, sex cells(gametes) carry only 23 chromosomes one chromosome from each homologous pair. Mendel, Johann Gregor = The father of genetics Mutation = A change in the DNA instructions. A change in the DNA sequence. The change can be beneficial, detrimental or neutral. It ultimately results in a change in protein. For instance, random genetic mutation gave rise to the dark phenotype of the peppered moth. Non-Disjunction: When homologous chromosomes fail to segregate properly during meiosis. Down syndrome, Turner syndrome and Klinefelter syndrome result from non-disjunction. Nucleotide = One of the four DNA chemical submits: (A + sugar and phosphate), (T + sugar and phosphate), (G + sugar and phosphate), (C + sugar and phosphate). 3 billion nucleotides make human genome Phenotype = the way an organism looks.( EXTERNAL CHARACTERISTICS) Recessive = A small, weaker allele is recessive. (CANNOT EXPRESS ITSELF IN HETEROZYGOUS CONDITION) Segregation = This is one of Mendels principles. Mendel said that all genes are comprised of 2 factors, one from each parent. Chromosomes segregate during meiosis. These factors (alleles) of a gene separate during the formation of gametes (sperm and egg). This ensures that each parent contributes 50% of their genetic information. Sex chromosomes = Chromosomes that determine sex (XY and XX) Somatic Cell = Body cell that contains 46 chromosomes in humans. Tay Sachs Disease = Autosomal recessive. Children die young. Head enlarges.... 60

Trait = is a feature of an organism. Uracil = in RNA, this nitrogen base matches with Adenine instead of Thymine.

5. Mutation
Mutations sudden change in DNA sequences resulting in changes in the phenotype and genotype Mutagens that causes mutation (X-ray, UV ray, harmful chemicals etc) Types of Mutations

Chromosomal mutation

Gene mutation

Structural

Numeral

Point

Frame shift

II

III

IV

I - Deletion II- Duplication (addition) Euploidy 61 Aneuploidy

III Inversion IV Translocation

i. triploid (3n)

i. Nullisomy

(2n-2)

ii. Tetraploid (4n) ii.Monosomy (2n-1) iii. Pentaploid (5n) iii Trisomy (2n+1) iv. Hexaploid (6n) iv Tetrasomy (2n+2) (POLYPLOIDY)

Aneupolidy is due to non disjunction (non separation) of chromosome during meiosis

GENETIC DISORDERS

62

Mendelian Disorder Single gene disorder Follow Mendellian principle of inheritance Gene disorders (gene mutation & point mutation)

Chromosomal disorder Involve a chromosome ,its part or complete set of chromosomes Do not follow Mendellian principle of inheritance Chromosomal aberrations (breakage deletion polyploidy and aneuploidy)

GENETIC DISORDERS

AUTOSOMAL Recessive Sickle cell anemia Phenylketonuria Albinism Dominant Polydactyly Huntingtons disease

X- CHROMOSOME LINKED Recessive Haemophilia Colour blindness Dominant Very rare e.g. disease Retts

CHROMOSOMAL DISORDERS AUTOSOMAL Downs syndrome (trisomy of chromosome number 21 ) total chromosome 47 X- CHROMOSOME LINKED Turners syndrome (absence of one X chromosome) 45,X0 Klinefelters syndrome (presence of an extra X chromosome) 47,e.g. XXY

Note: I n case of X chromosome linked chromosomal disorders (aneuploidy) Presence of Y chromosome phenotype is male Absence of Y chromosome phenotype is female

63

Chapter 6. MOLECULAR BASIS OF INHERITANCE

DNA is a polynucleotide. When many nucleotides are linked with each other in a linear fashion, the resulting chain, is referred to as a polynucleotide (polymer). For example, each strand of DNA molecule has many deoxyribonucleotides linked in a chain-like arrangement. Therefore, it is described as polynucleotide strand and DNA molecule as polynucleotide molecule. In the DNA polynucleotide strands, the nucleotides are joined with each other by phosphodiester linkages (Figure. 1.). In a phosphodiester linkage, the phosphate group present at the C-5 of the sugar of one nucleotide gets attached to the C-3 of the sugar of the next nucleotide in the chain. (b) The Watson-Crick model of the DNA double helix : The most widely accepted model for the structure of DNA molecule (Figure 8.4) was proposed by Watson and Crick in 1953 (who won the Nobel Prize for Medicine in 1962). According to their model, the DNA has the following structural characteristics. i. Molecule : The DNA molecule is a double helix (Fig. 1A). The molecule is formed by two antiparallel polynucleotide strands which are spirally coiled around each other in a right-handed helix. The two strands are held together by hydrogen bonds. The double stranded helical molecule has alternate major (or deep) and minor grooves.

Fig 8.4

Figure 1. Structure of DNA (Watson and Crick model) (A) DNA double helix. (B) Detailed structure of the two strands. (C) C-5 and C-3 ends and antiparallel nature of strands (diagrammatic) ii. Structure of each strand (8.4 B) : Each strand is a long polynucleotide of deoxyribonucleotides. The backbone of the strand is formed by alternately arranged deoxyribose sugar and phosphate molecules 64

which are joined by the phosphodiester linkages. Each sugar in the strand has one base horizontally attached to it at carbon-1. It can be any one of the four: A,T,G,or C. These four N-bases can occur in any possible sequence along the length of a strand. iii. Complementary nature of the strands : The two strands are complementary to each other with regards to the arrangement of the bases in the two strands. For example, where adenine (a purine) occurs in one strand, thymine (a pyrimidine) is present in the corresponding position in the opposite strand and vice versa. Similarly, wherever guanine (a purine) is present in one strand, the other strand has cytosine (a pyrimidine) opposite to it and vice versa. Thus, in the double helix, purines and pyrimidines exist in base pairs, i.e., (A and T) and (G and C). As a result, if the base sequence of one strand of DNA is known, the base sequence of its complementary strand can be easily deduced. vi. Complementary base pairing : In each pair, the two bases of the opposite strands are joined by hydrogen bonds. A and T are joined by two hydrogen bonds, while G and C are joined by three hydrogen bonds. This is called complementary base pairing. The two strands are thus held together all along their lengths by these hydrogen bonds. v. Purine : Pyrimidine ratio : Because of the fixed or complementary base pairing in the DNA molecule, the total number of A is equal to the total number of T and the total number of G is equal to the total number of C. In other words, (A+G)= (T+C). Hence, purines: pyrimidines ratio is 1:1. vi. C-3 and C-5 ends of the strand : In each strand one end of the strand has one free phosphate group on carbon-5 of the sugar molecule. This is the end of the strand is called C-5 (or 5') end. The other end of the strand has a free -OH on carbon-3 of the sugar molecule. This is called C-3 (or 3') end of the strand (Figure 1c). vii. Antiparallel nature of strands : The two strands are oppositely oriented and hence are called antiparallel. This means, the 3' end of one strand is adjacent to the 5' end of the other strand (Fig. 8.4c) . This is because, the phosphate-sugar linkages run in opposite directions in the two strands. viii. Dimensions: The diameter of the DNA double helix is 20 Ao. The length of each complete spiral (turn or pitch) of the molecule measures 34 Ao. 10 pairs of nucleotides are present in each complete spiral. Therefore, each nucleotide in the strand occupies a distance of 3.4A0.

(c) Circular DNA molecules : Chromosomes of most prokaryotes (e.g. bacteria, cyanobacteria,) are circular molecules of DNA. In bacteria, there is no organized nucleus. The bacterial nucleoid consists of a single circular DNA molecule (bacterial chromosome). The molecule has two complementary strands forming a covalently closed circle. Generally, the circular molecule is present in a highly folded and suspercoiled state (Figure 8.5A). This is expected because the diameter of a bacterial cell (e.g. Escherichia coli) is about 1-2 microns while the total length of the circular DNA is about 1100 microns. The circular molecule has 40-50 folds or looped domains. These folds are held in position by RNA molecules (RNA connectors) and some nonhistone proteins associated with the bacterial chromosome. (Histones are absent in bacteria The DNA segment in each loop is supercoiled independently. Because of this characteristic formation of loops as well as the supercoils within the loops, the large circular DNA molecule can be packed into a 65

small bacterial cell. Otherwise, the relaxed and fully expanded circular molecule (Figure 8.5) would be too large (about 350 microns diameter) to be contained in the bacterial cell. The coils of the supercoiled circle can be relaxed by treatment with enzymes such as RNAse or DNAse. The uncoiling occurs due to a break (nick) in one or both the strands of DNA. In some viruses, e.g. certain bacteriophages, the circular DNA is single stranded. It becomes double stranded only during replication (replicatable form).

8.1 Packaging of Hereditary Material

Structure of nucleus (A) in a prokaryotic cell and (B) in an eukaryotic cell REPLICATION:The various steps involved in this process are summarized as follows: i. ii. iii. iv. v. vi. The mechanism of replication starts at a specific point of the DNA molecule. This is called the origin of replication (orisite) At the origin, the DNA strand breaks because of an incision (nick). This is made by an enzyme called incision enzyme (endonuclease). The hydrogen bonds joining the two strands are broken by the enzyme. The two strands start unwinding. This takes place with the help of a DNA unwinding protein. The two polynucleotide strands are thus separated. The point where the two strands separate appears like a fork or a y-shape. This is described as a replicating fork. A new strand is constructed on each old strand. This takes place with the help of a small RNA primer molecule which is complimentary to the DNA at that point. Each old DNA strand acts as a template (site) for the construction of new strand. The RNA primer attaches itself to the old strand and attracts the enzymes which add new nucleotides. The deoxyribose nucleotides are present in the surrounding nucleoplasm. Appropriate nucleotides are selected from the nucleoplasm, and are attached by H-bonds to their respective complementary bases on the old strand. A new DNA strand is thus constructed opposite to each old strand 66

As stated earlier, chromosomes are the carriers of the hereditary material. In cells, chromosomes are located in the nucleus. In prokaryotic cells (e.g., bacteria, cyanobacteria, etc.), organized nuclei are not present. As a result, the hereditary material occurs in the cell cytoplasm as a nucleoid (Figure 8.1-A) without the nuclear envelope. (a) Eukaryotic nucleus (Figure 8.1-B): In eukaryotic cells (e.g., all higher plants and animals), a well organized nucleus is present. It is bounded by a double membrane nuclear envelope with trilaminar structure. There are pores in the envelope. Internally, the nucleus is filled with nucleoplasm (nuclear sap) that is acidophilic and clear. The interphasic nucleus (i.e. the nucleus which is not in the stage of division) usually shows the presence of one dark, spherical body called nucleolus. The nucleoplasm also contains nuclear reticulum or chromatin network. It consists of very fine, long chromatin fibers. These represent the chromosomes at the interphasic stage. They become short, thick and distinct during cell division. (b) Structure of chromatin (Figure 8.2): The chromosomes are composed of nucleoprotein called chromatin. The chromatin fiber appears to have a structure like a string of beads. According to the nucleosome-solenoid model proposed by Kornberg and Thomas (1974), the beaded string is made of repeating units called nucleosomes (Figure 8.2A). Each nucleosome is a bead-like particle about 60 Ao high and 110 Ao wide.

Figure 8.2. Nucleosome - solenoid model for the structure of chromatin In the nuclesome, the DNA double helix is wound around the core of eight histone molecules (octamer). The segments of DNA joining the beads are called linker DNA. Each linker DNA has an average of one molecule of a histone protein (H1) attached to it. The thin, long and "beads-on-string" chromatin fiber is condensed and packed into a short and thick chromosome (metaphase chromosome) because of further coiling and "super coiling", wherein (1) the string of beads condenses and coils to produce a 100 Ao thick nucleosome fiber (2) the nucleosome fibre then supercoils to produce a 300-350 Ao thick solenoid fiber. A solenoid represents the structure of chromatin fiber in the metaphase chromosome. 8.2 The Structure of DNA

67

Deoxyribonucleic nucleic Acid (DNA) is a highly complex megabiomolecule. The long chain molecule is formed of repeating units called nucleotides. Hence, it is described as the polynucleotide molecule. It consists of two polynucleotide antiparallel strands which are spirally coiled round each other along their lengths (Watson and Crick, 1953). (a) Chemical Components of DNA : The highly complex DNA molecule is composed of only three types of chemical components. These are (i) deoxyribose sugar, (ii) a phosphate, and (iii) nitrogen containing organic bases. i. Deoxyribose sugar : The sugar present in the DNA molecule is called deoxyribose sugar and hence the nucleic acid is called deoxyribonucleic acid (DNA). It is a pentose sugar (with 5 carbon atoms) having a pentagonal ring structure (Figure 8.3) Phosphate (Figure 8.3) : The phosphate in the DNA is present as phosphoric acid (H3PO4). It has three reactive (-OH) groups of which two are involved in the formation of the sugar-phosphate backbone of each DNA strand. The nitrogen-containing organic bases

ii.

iii.

These are heterocyclic compounds containing nitrogen in their rings and therefore called nitrogenous bases. DNA contains four different bases called adenine (A), guanine (G) cytosine (C), and thymine (T). These are grouped into two classes on the basis of their chemical structure: (i) Purines (with a double ring structure) and (ii) Pyrimidines (with a single ring structure) (Figure 8.3) Nucleosides : In the molecules of nucleic acids (DNA and RNA), each pentose sugar molecule has one nitrogen base attached at carbon number 1. It may be either a purine or a pyrimidine base. Thus, a pentose sugar with the attached N-base forms a nucleoside. [Sugar + N-base] = Nucleoside. In DNA, the deoxyribose sugar has one of the four bases (A,G,T or C) attached. Therefore, the nucleosides in DNA are called deoxyribosides. [deoxyribose sugar + N-base] = Deoxyriboside Nucleotides : A nucleoside with a phosphate group attached to it is called a nucleotide

68

Fig.8.3 Chemical components of DNA and the phosphodiester linkage

. [Nucleoside + Phosphate] = Nucleotide A nucleotide is the basic unit or monomer in the structure of a nucleic acid molecule. Thus, a nucleotide is a nucleoside phosphate. In a nucleotide, the phosphate group is linked with the pentose sugar at carbon-5. In the DNA, each nucleotide is called deoxyribonucleotide. [Deoxyribose sugar + N-base + Phosphate] = Deoxyribonucleotide As there are four different bases (A,T,G and C) in DNA, there can be only four types of nucleotides in DNA. The nucleotides act as the building block molecules for the synthesis of the polynucleotide molecules. When two nucleotides are linked together, a dinucleotide (or dimer) results. 69

8. 3 Replication Of DNA In Eukaryotes Definition: "The process by which DNA produces daughter DNA molecules which are exact copies of the original DNA is called replication of DNA." In eukaryotes, DNA is double stranded. The two strands are complementary to each other because of their base sequences. Semi-conservative method of DNA replication Important points: (i) This is the most common method of DNA replication. (ii) It takes place in the nucleus where the DNA is present in the chromosomes. (iii) Replication takes place in the S-phase (synthesis phase) of the interphase nucleus. (iv) The deoxyribose nucleotides needed for the formation of the new DNA strands are present in the nucleoplasm. At the time of replication, the two strands of DNA first separate. Each strand then acts as a template for the formation of a new strand. A new strand is constructed on each old strand, and two exactly identical double stranded DNA molecules are formed. In each new DNA molecule, one strand is old (original) while the other is newly formed. Hence, Watson and Crick described this method as semi-conservative replication.

(A) An overall process of DNA replication showing replication fork and formation of new strands template and lagging template

70

Figure Stages in the semi-conservative method of DNA replication in eukaryotes The various steps involved in this process are summarized as follows: i. The mechanism of replication starts at a specific point of the DNA molecule. This is called the origin of of replication (ori site) ii. At the origin, the DNA strand breaks because of an incision (nick). This is made by an enzyme called incision enzyme (endonuclease). iii. The hydrogen bonds joining the two strands are broken by the enzyme. iv. The two strands start unwinding. This takes place with the help of a DNA unwinding v. enzyme(unwindase) Helicases. The two polynucleotide strands are thus separated. v. The point where the two strands separate appears like a fork or a Y-shape. This is described as a replicating fork. vi. A new strand is constructed on each old strand. This takes place with the help of a small RNA primer molecule which is complimentary to the DNA at that point. Each old DNA strand acts as a template (site) for the construction of new strand. The RNA primer attaches itself to the old strand and attracts the enzymes(DNApolymeraseIII) which add new nucleotides through base complementation. The deoxyribose nucleotides are present in the surrounding nucleoplasm.. A new DNA strand is thus constructed opposite to each old strand vii. The formation of new complementary strand always begins at the 3' end of the template strand (original strand) and progresses towards the 5' end (i.e in 3' - 5' direction). Since the new strand is antiparallel to the template strand, it is obvious that the new strand itself is always developed in the, 5'-3' direction. For this reason when the two original strands separate (then with respect to the origin of separation), one acts as 3'-5' template while the other acts as 5'- 3' template. Of the two, the replication of 3'-5' template begins first. Hence the new strand formed on it is called the leading strand. The other template (5'-3') must begin replication at the fork and progress back toward the previously transcribed fragment. The new strand formed on it is called the lagging strand. Replication of the lagging strand takes place in small fragments called Okazaki fragments. These are then connected together by the enzyme ligase. Replication may take place in only one direction on the DNA helix (unidirectional) or in two directions (bidirectional). At the end of the process, two double stranded DNA molecules are formed from the original DNA molecule. 71

viii. ix. x.

xi

In each newly formed DNA, one strand is old(parental) while the other is new. Hence, it is described as semi-conservative replication.

It is very important to know that DNA replication is not a passive and spontaneous process. Many enzymes are required to unwind the double helix and to synthesize a new strand of DNA. We will approach the study of the molecular mechanism of DNA replication from the point of view of the machinery that is required to accomplish it. The unwound helix, with each strand being synthesized into a new double helix, is called the replication fork.

The Enzymes of DNA Replication

Topoisomerase is responsible for uncoiling of double halix. The tension holding the helix in its coiled and supercoiled structure can be broken by nicking a single strand of DNA. Try this with string. Twist two strings together, holding both the top and the bottom. If you cut only one of the two strings, the tension of the twisting is released and the strings untwist. Helicase accomplishes unwinding of the original double strand, once supercoiling has been eliminated by the topoisomerase. The two strands very much want to bind together because of their hydrogen bonding affinity for each other, so the helicase activity requires energy (in the form of ATP ) to break the strands apart. DNA polymerase proceeds along a single-stranded molecule of DNA, recruiting free dNTP's (deoxy-nucleotide-triphosphates) to hydrogen bond with their appropriate complementary dNTP on the single strand (A with T and G with C), and to form a covalent phosphodiester bond with the previous nucleotide of the same strand. The energy stored in the triphosphate is used to covalently bind each new nucleotide to the growing second strand. There are different forms of DNA polymerase , but it is DNA polymerase III that is responsible for the processive synthesis of new DNA strands. DNA polymerase cannot start synthesizing de novo on a bare single strand. It needs a primer with a 3'OH group onto which it can attach a dNTP. DNA polymerase is actually an aggregate of several different protein subunits, so it is often called a holoenzyme. The holoenzyme also has proofreading activities, so that it can make sure that it has inserted the right base, and nuclease (excision of nucleotides) activities so that it can cut away any mistakes it might have made. Primase is actually part of an aggregate of proteins called the primeosome. This enzyme attaches a small RNA primer to the single-stranded DNA to act as a substitute 3'OH for DNA polymerase to begin synthesizing from. This RNA primer is eventually removed by RNase H and the gap is filled in by DNA polymerase I. Ligase can catalyze the formation of a phosphodiester bond given an unattached but adjacent 3'OH and 5'phosphate. This can fill in the unattached gap left when the RNA primer is removed and filled in. The DNA polymerase can organize the bond on the 5' end of the primer, but ligase is needed to make the bond on the 3' end. Single-stranded binding proteins are important to maintain the stability of the replication fork. Single-stranded DNA is very labile, or unstable, so these proteins bind to it while it remains single stranded and keep it from being degraded.

72

The

Steps

of

DNA

Replication

A portion of the double helix is unwound by a helicase. A molecule of DNA polymerase binds to one strand of the DNA and begins moving along it in the 3' to 5' direction, using it as a template for assembling a leading strand of nucleotides and reforming a double helix. Because DNA synthesis can only occur 5' to 3', a second DNA polymerase molecule is used to bind to the other template strand as the double helix opens. This molecule must synthesize discontinuous segments of polynucleotides (called Okazaki fragments). Another enzyme, DNA ligase then stitches these together into the lagging strand

Speed

of

Replication

Prokaryotes - The single molecule of DNA that is the E. coli genome contains 4.7 x 106 nucleotide pairs. DNA replication begins at a single, fixed location in this molecule, the replication origin, proceeds at about 1000 nucleotides per second, and thus is done in no more than 40 minutes. And thanks to the precision of the process (which includes a "proof-reading" function), the job is done with only about one incorrect nucleotide for every 109 nucleotides inserted. In other words, more often than not, the E. coli genome (4.7 x 106) is copied without error! Eukaryotes - The average human chromosome contains 150 x 106 nucleotide pairs which are copied at about 50 base pairs per second. The process would take a month (rather than the hour it actually does) but for the fact that there are many replication origins on the eukaryotic chromosome. Replication begins at some origins earlier in S phase than at others, but the process is completed for all by the end of S phase. As replication nears completion, "bubbles" of newly replicated DNA meet and fuse, finally forming two new molecules

. The semi-conservative nature of DNA replication was confirmed by Meselson and Stahl (1958) with the help of an experiment. They marked the DNA in Esherichia coli with heavy isotope of nitrogen (15N) and then traced it in the following generations of the E-coli progeny. It was demonstrated that each daughter DNA has one strand of the parent (old strand) and the other strand in newly formed. 73

Nitrogen is a major constituent of DNA. 14N is by far the most abundant isotope of nitrogen, but DNA with the heavier (but non-radioactive) 15N isotope is also functional. E. coli were grown for several generations in a medium with 15N. When DNA is extracted from these cells and centrifuged on a salt(CesiumChloride) density gradient, the DNA separates out at the point at which its density equals that of the salt solution. The DNA of the cells grown in 15 N medium had a higher density than cells grown in normal 14N medium. After that, E. coli cells with only 15N in their DNA were transferred to a 14N medium and were allowed to replicate; the progress of cell division was monitored by measuring the optical density of the cell suspension. DNA was extracted periodically and was compared to pure 14N DNA and 15N DNA. After one replication(20minutes), the DNA was found to have close to the intermediate density. Semiconservative replication would result in double-stranded DNA with one strand of 15N DNA, and one of 14N DNA.
74

The authors continued to sample cells as replication continued. DNA from cells after two replications(after 40minutes) had been completed was found to consist of equal amounts of DNA with two different densities, one corresponding to the intermediate density of DNA of cells grown for only one division in 14N medium, the other corresponding to DNA from cells grown exclusively in 14N medium. The result was consistent with the semiconservative replication hypothesis. 8.6 RNA : Structure and Types Ribonucleic Acid (RNA) is another polynucleotide which occurs in the cells as non-genetic material, with the exception of some viruses. RNA is present in the nucleus as well as in the cytoplasm.

General structure RNA molecule is single stranded and consists of nucleotides arranged in a long series. The single strand of RNA may be simple and straight, or it may be variously folded upon itself in certain regions. Structural components : RNA molecule has three primary components. 1. Ribose sugar (a pentose sugar), with a pentagonal ring structure 2. Phosphate, as phosphoric acid 3. Nitrogenous bases There are four kinds of nitrogenous bases found in RNA. Of those, two are purines and two are pyrimidines, as follows :

Thus in RNA, uracil is present in place of thymine found in DNA. Structure of an RNA strand : The strand is made up of alternating molecules of ribose sugar and the phosphate. The nitrogen bases are attached to the sugar molecules in the strand and stick out laterally as in DNA (figure 8.9). A sugar, a N-base and a phosphate together form a ribonucleotide. A nucleotide without the phosphate is called ribonucleoside. RNA being single stranded, the nitrogen bases remain mostly unpaired. However, the strand may be folded upon itself in certain regions. In such folded regions, base pairing occurs between purines and pyrimidines as follows :

75

Adenine = Uracil (two H-bonds). Guanine Cytosine (three H-bonds). Nitrogen bases remain unpaired in the unfolded regions of the strand. Because of this variability in base pairing in different regions of the same strand, the total number of purines need not be equal to the total number of pyrimidines in RNA

Types of non-genetic RNA and their functions There are three types of non-genetic RNA. (1) mRNA or messenger RNA, (2) rRNA or ribosomal RNA and (3) tRNA or transfer RNA

(1) mRNA (Messenger RNA) : This is called messenger RNA because it carries information for protein synthesis from the DNA to the ribosomes in the cytoplasm (the site of protein synthesis). mRNA constitutes about 3-5% of the total RNA. It is produced on the DNA strand. The process is called transcription. Hence, the base sequence of mRNA is complementary to that of the DNA strand. The bases on the mRNA strand are organized into triplets. Each triplet consists of a sequence of three consecutive bases (nucleotides) and is called a codon (code word). Each codon specifies one amino acid. The sequence of codons on the mRNA strand is called the mRNA language. It indicates the sequence of amino acids for the synthesis of a protein. It begins with the codon AUG (initiation codon or starting codon) and ends with either UAA, UAG or UGA (stop codons). The single-stranded mRNA molecule is always straight (Fig. 8.10) and therefore, base pairing is totally absent in mRNA.

Role of m-RNA in protein synthesis i. ii. iii. Represents the sequence of codons from the DNA strand (transcription). Brings the sequence to the ribosomes (site of protein synthesis) in the cytoplasm. Provides the sequence for the synthesis of specific protein from the amino acids with the help of tRNA (translation).

(2) rRNA (Ribosomal RNA) : r RNA forms about 80% of the total RNA. It is present in the ribosomes in the cell cytoplasm (site of protein synthesis) and hence called rRNA. The single-stranded molecule of rRNA is variously folded and twisted upon itself in certain regions (Figure 8.11). In such folded regions, complementary bases form pairs and are joined by hydrogen bonds. Role of rRNA in protein synthesis : The role of rRNA in protein synthesis is not yet very clearly known but it is known to complex with various protiens. The resulting structure is a ribosome, and this complex reads the coded sequence in mRNA to link amino acids together into particular protiens. i. ii. iii. It provides proper binding sites for the mRNA of the ribosomes. It orients the mRNA in such a way that its nitrogen base triplets or codons are properly read or translated. It also releases tRNA after the transfer of activated amino acid. 76

iv. v.

It protects the mRNA strand from the action of enzymes (nucleases). It protects the growing (nascent) polypeptide chain from proteolytic enzymes.

(3) tRNA (Transfer RNA) : It is the smallest of all the types of RNA. About 10 to 20% of the total RNA of the cell is of this type. tRNA strand is folded upon itself forming loops. It results in either a clover leaf pattern or hair pin pattern (Figure 8.12). One end of the strand has guanine, while the other end carries the CCA combination of nitrogen bases. A triplet of nitrogen bases called anticodon is present on one of the loops. The anticodon pairs with the complementary codon on the mRNA molecule. The tRNA molecules carry amino acids to the mRNA during the process of protein synthesis. Each type of the amino acid is carried by a specific tRNA molecule. tRNA is synthesized on the DNA template. It has complementary base pairs in folded regions. Role of tRNA in protein synthesis i. ii. iii. tRNA carries the required specific amino acids from cell cytoplasm to the ribosome (site of protein synthesis). Each type of amino acid is carried by a specific type of tRNA. In the ribosome, tRNA helps to arrange the amino acids in their proper sequence for the synthesis of a protein. This is done with the help of the codons on the mRNA and the matching (complementary) anticodons on the tRNA (translation).

CLOVERLEAF STRUCTURE OF tRNA Table 8.2 : Difference between DNA and RNA Characters 1. Molecule DNA Double stranded, helical RNA Single stranded, straight or variously folded and twisted. Ribose Uracil Normally absent,

2. 3. 4.

Pentose sugar Pyrimidine base Complementary

Deoxyribose Thymine Always present

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base pairing

and exists but may be between A = T present in twisted and G = C segments of a molecule. If present, pairing is between A = U and G = C

5.

Ratio of Purines: Pyrimidines Replication

Always 1 : 1

Not 1:1

necessarily

6.

Can replicate

Cannot replicate

CLOVERLEAF STRUCTURE OF tRNA

The main principle of protein synthesis. According to Crick (1958), DNA determines the sequence of amino acids in a polypeptide (protein) through mRNA. This is the main principle (central dogma) of protein synthesis. This involves transcription and translation.

Components involved in protein synthesis : The process requires 1. 2. 3. 4. 5. The 20 types of amino acids that are specified by the genetic code. DNA (to provide the base sequence). The non-genetic RNAs (m-RNA, t-RNA and r-RNA). Ribosomes (site of protein synthesis) and Various enzymes (f0actors).

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Mechanism of protein synthesis As stated earlier, the mechanism of protein synthesis involves two main events namely (A) Transcription (B) Translation. (A) Transcription Transcription is the process of formation of mRNA on the DNA strand. It takes place in the presence of enzyme RNA Polymerase. One of the strands of DNA acts as a template for the formation of the same m-RNA in a series, one by one. Such ribosomes bound to the same mRNA strand are called polyribosomes or polysomes. The polypeptide chain formation in each ribosome is independent and the same as described above.

Transcription is the first stage of the expression of genes into proteins. In transcription, a mRNA (messenger RNA) intermediate is transcribed from one of the strands of the DNA molecule. The RNA is called messenger RNA because it carries the 'message' or genetic information from the DNA to the ribosomes, where the information is used to make proteins. RNA and DNA use complementary coding, where base pairs match up, similar to how the strands of DNA bind to form a double helix. One difference between DNA and RNA is that RNA uses uracil in place of the thymine used in DNA. RNA polymerase mediates the manufacture of an RNA strand that complements the DNA strand. RNA is synthesized in the 5' -> 3' direction (as seen from the growing RNA transcript).

The Transcription Process

RNA synthesis involves separation of the DNA strands and synthesis of an RNA molecule in the 5' to 3' direction by RNA polymerase, using one of the DNA strands as a template. In complementary base pairing, A, T, G, and C on the template DNA strand specify U, A, C, and G, respectively, on the RNA strand being synthesized.

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In a prokaryotic cell, transcription and translation are coupled; that is, translation begins while the mRNA is still being synthesized. In a eukaryotic cell, transcription occurs in the nucleus, and translation occurs in the cytoplasm. Prokaryotic Cell

Because there is no nucleus to separate the processes of transcription and translation, when bacterial genes are transcribed, their transcripts can immediately be translated. Eukaryotic Cell

Transcription and translation are spatially and temporally separated in eukaryotic cells; that is, transcription occurs in the nucleus to produce a pre-mRNA molecule. The pre-mRNA is typically processed to produce the mature mRNA, which exits the nucleus and is translated in the cytoplasm

mRNA in Eukaryotes
The sequence of a eukaryotic protein-coding gene is typically not colinear with the translated mRNA; that is, the transcript of the gene is a molecule that must

80

be processed to remove extra sequences (introns) before it is translated into the polypeptide.

Most eukaryotic protein-coding genes contain segments called introns, which break up the amino acid coding sequence into segments called exons. The transcript of these genes is the pre-mRNA (precursor-mRNA).

The pre-mRNA is processed in the nucleus to remove the introns and splice the exons together into a translatable mRNA. That mRNA exits the nucleus and is translated in the cytoplasm.

Pre-mRNA Processing (Splicing)

Eukaryotic pre-mRNAs typically include introns. Introns are removed by RNA processing in which the intron is looped out and cut away from the exons by snRNPs, and the exons are spliced together to produce the translatable mRNA.

81

The steps of pre-mRNA splicing (intron removal) are as follows: The intron loops out as snRNPs (small nuclear ribonucleoprotein particles, complexes of snRNAs and proteins) bind to form the spliceosome. The intron is excised, and the exons are then spliced together.

The resulting mature mRNA may then exit the nucleus and be translated in the cytoplasm.

(B) Translation In Translation one particular group gets transmitted from one Band of DNA to another Band of DNA. In this procedure the DNA polymer (enzyme) is used. Normally, 40% of protein synthesis occurs in this manner. The genetic code : DNA is genetic material and contains genetic information. The expression of a gene takes place through specific enzymes. Each gene produces a specific (one-gene one-protien hypothesis). In other words, formation of each specific protein is controlled by a particular gene. A gene is (almost always) a segment of DNA strand and so, the information for the formation of a protein is contained in the DNA strand. Further, each protein is a long polypeptide chain molecule formed by joining amino acid molecules. From the cell pool, only 20 different types of amino acids are used for protein synthesis. The sequence of the nitrogen bases in the DNA determines the sequence of amino acids in a protein molecule through the mRNA. This sequence is copied down by the mRNA (transcription). It is present on

82

the mRNA strand in the form of coded language (cyptogram or mRNA language or genetic code). The mRNA bases (A, U, C and G) serve as the four alphabets of the coded language. Codon : The smallest sequence of the nitrogen bases (nucleotides) on the mRNA which can specify one amino acid is called a codon . Each codon consists of three successive bases on the mRNA. Why should each codon in the genetic code consist of 3 bases (triplet codon) and not of one base each or 2 bases each? This is because there are 20 different amino acids which can be used in the synthesis of proteins in the cells. There must be at least one specific codon for each amino acid. Thus, there has to be at least 20 different codons in the genetic code. There are only four bases. A minimum of 3 bases per codon is necessary to have (a minimum of) 20 codes. The wobble hypothesis (Crick, 1966) : The anticodon on tRNA is complementary to the codon on the mRNA as per the A = U, G = C base pairing rule. However, it has been observed that the 3rd base position may vary and yet still code for the same amino acid. For example, both codons TTA and TTG code for the amino acid, leucine. Thus, the third position is called the wobble position. Thus, Cricks (1966) wobble hypothesis explains the degeneracy of the genetic code at the third position of the codon. 8.8 The Central Theme of Protein Synthesis A protein is a long chain polypeptide. The chain molecule is formed by joining amino-acid molecules with peptide linkages. The sequence of amino acids in each type of protein is highly specific. The functional proteins form enzymes and hormones and control all metabolic and biochemical reactions and processes in cells. Any alteration in the sequence of amino acids in protein molecules can affect the function of that protien. Hence, bio-synthesis of proteins is a highly specific process.

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SUMMARY : MOLECULAR GENETICS AND GENE EXPRESSION (1) DNA is the genetic material in almost all organisms. (2) Chromosomes carry genes. (3) The chromatin consists of DNA and two types of proteins (histones and non-histones). (4) During prophase of the cell division, DNA of the chromatin undergoes coiling, super coiling and folding and along with the associated proteins, forms highly a condensed and prominent chromosome (as seen during metaphase). (5) This is explained with the help of necleosome solenoid model (Komberg and Thomas, 1974). (6) DNA molecule has a double stranded, helical structure in which the two antiparallel polynucleotide strands are helically coiled round each other, and joined by hydrogen bonds (Watson and Crick). (7) DNA has the ability of self-replication. (8) RNA molecule is single stranded. It can not replicate. It is produced on DNA strand. (9) Non-genetic RNA is of 3 types and helps in protein synthesis. (10) In prokaryotes, the single circular chromosome has a circular DNA capable of replication. (11) Prokaryotes also possess plasmids and episomes. These are the extra chromosomal circular DNA which carry genes and are capable of selfreplication. (12) Genetic information required to produce a protein is called the genetic code. It is present on the DNA in the form of the base sequence. It is transcribed on m-RNA and is then used for synthesis of protein in the ribosomes. (13) Genetic code is triplet. (14) Each triplet or codon on m-RNA specifies one amino acid. (15) In all, there are 64 codons in the genetic code of which 61 codons specify 20 different amino acids. (16) Three codons (UAA, UAG and UGA) do not specify any acid (stop codons). (17) AUG is the starting or initiation codon. (18) In protein synthesis, m-RNA provides the sequence of the arrangement of amino acids in form of the codon sequence. These are carried by the t-RNA and are arranged in the sequence with the help of matching anticodons on t-RNA. (19) After formation, the peptide 84

chain is released with the help of release factors.

Griffith's Transformation Experiment

Griffith's experiment, conducted in 1928 by Frederick Griffith, was one of the first experiments suggesting that bacteria are capable of transferring genetic information through a process known as transformation. Griffith used two strains of Diplococcus pneumoniae (which infects mice), a type III-S (smooth) and type II-R (rough) strain. The III-S strain covers itself with a polysaccharide capsule that protects it from the host's immune system, resulting in the death of the host, while the II-R strain doesn't have that protective capsule and is defeated by the host's immune system. A German bacteriologist, In this experiment, bacteria from the III-S strain were killed by heat, and their remains were added to II-R strain bacteria. While neither alone harmed the mice, the combination was able to kill its host. Griffith was also able to isolate both live II-R and live III-S strains of pneumococcus from the blood of these dead mice. Griffith concluded that the type II-R had been "transformed" into the lethal III-S strain by a "transforming principle" that was somehow part of the dead III-S strain bacteria. Today, we know that the "transforming principle" Griffith observed was the DNA of the III-S strain bacteria. While the bacteria had been killed, the DNA had survived the heating process and was taken up by the II-R strain bacteria. The III-S strain DNA contains the genes that form the protective polysaccharide capsule. Equipped with this gene, the former II-R strain bacteria were now protected from the host's immune system and could kill the host. The exact nature of the transforming principle (DNA) was verified in the experiments done by Avery, McLeod and McCarty and by Hershey and Chase. The AveryMacLeodMcCarty experiment was an experimental demonstration, reported in 1944 by Oswald Avery, Colin MacLeod, and Maclyn McCarty, that DNA is the substance that causes bacterial transformation. It was the culmination of research in the 1930s and early 1940s at the Rockefeller Institute for Medical Research to purify and characterize the "transforming principle" responsible for the transformation phenomenon first described in Griffith's experiment of 1928: killed Streptococcus pneumoniae of the virulent strain type III-S, when injected along with living but non-virulent type II-R pneumococci, resulted in a deadly infection of type III-S pneumococci. In their paper "Studies on the Chemical Nature of the Substance Inducing Transformation of Pneumococcal Types: Induction of Transformation by a Desoxyribonucleic Acid Fraction Isolated from Pneumococcus Type III", published in 85

the February 1944 issue of the Journal of Experimental Medicine, Avery and his colleagues suggest that DNA, rather than protein as widely believed at the time, may be the hereditary material of bacteria, and could be analogous to genes and/or viruses in higher organisms.[

To show that it was DNA rather than some small amount of RNA, protein, or some other cell component that was responsible for transformation, Avery and his colleagues used a number of biochemical tests. They found that trypsin, chymotrypsin and ribonuclease (enzymes that break apart proteins or RNA ) did not affect it, but an enzyme preparation of "deoxyribonucleodepolymerase" (a crude preparation, obtainable from a number of animal sources, that could break down DNA ) destroyed the extract's transforming power Schematic Representation: Two strains of bacteria - Rough (R) and Smooth (S) o Rough - bacteria had a rough appearance in culture, non-virulent (doesn't kill) o Smooth - bacteria had a smooth appearance in culture, virulent (kills) Performed several experiments whose results can be summarized as follows: o Mouse + Live S >> Died. Autopsy revealed infestation of S strain bacteria o Mouse + Live R >> Lived a happy life These two experiments illustrate the effects of normal R and S strains o Mouse + Heat-killed S >> Lived a happy life o Mouse + Heat-killed R >> Lived a happy life These two experiments illustrate that dead bacteria by themselves are harmless o Mouse + Heat-killed S + Live R >> Died. Autopsy revealed infestation of S strain o Apparently something from the heat-killed S changed the live R to make them virulent this was called transformation Griffith didn't know what it was that transformed the R strain into the S strain, but he demonstrated that it could be done Oswarld Avery, Maclyn McCarty, and Colin MacLeod (1944) Performed the same experiment, only they used tissue cultures growing in petri dishes o Heat-killed S + Live R + protease >> Live R and S colonies o Heat-killed S + Live R + DNAse >> No growth

Other experiments - purified each chemical class associated with transformation and subjected bacteria to each one individually o Only DNA produced transformation

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Hershey and Chase conducted their experiments on the T2 phage, a virus whose structure had recently been shown by electron microscopy.The phage consists of a protein shell containing its genetic material. The phage infects a bacterium by attaching to its outer membrane and injecting its genetic material and leaving its empty shell attached to the bacterium.

In their first set of experiments, Hershey and Chase labeled the DNA of phages with radioactive Phosphorus-32 (the element phosphorus is present in DNA but not present in any of the 20 amino acids from which proteins are made). They allowed the phages to infect E. coli, and through several elegant experiments were able to observe the transfer of P32 labeled phage DNA into the cytoplasm of the bacterium. In their second set of experiments, they labeled the phages with radioactive Sulfur-35 (Sulfur is present in the amino acids cysteine and methionine, but not in DNA). Following infection of E. coli they then sheared the viral protein shells off of infected cells using a high-speed blender and separated the cells and viral coats by using a centrifuge. After separation, the radioactive S35 tracer was observed in the protein shells, but not in the infected bacteria, supporting the hypothesis that the genetic material which infects the bacteria was DNA and not protein.

o o

Bacteriophage - composed only of protein coat and DNA - injected something inside host bacterial cell which contained its genetic information Was the injected material DNA or protein?

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In the first experiment, illustrated on the left, the "radioactive" substance was the protein coat. In the second experiment, illustrated on the right, the "radiopactive" substance was the DNA. After the bacteriophage had injected the genetic material into the cell, the solutions were centrifuged and each reaction flask was measured for radioactivity. In the first experiment, all of the radioactivity was in the fluid that the cells were originally suspended. In the second experiment, all of the radioactivity was in the bacteria. It was concluded that since the radioactivity in the second experiment was in the bacteria, the genetic information was contained in the DNA and not the protein.

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What Are ESTs and How Are They Made?

An Expressed Sequence Tag is a tiny portion of an entire gene that can be used to help identify unknown genes and to map their positions within a genome.

ESTs are small pieces of DNA sequence (usually 200 to 500 nucleotides long) that are generated by sequencing either one or both ends of an expressed gene. The idea is to sequence bits of DNA that represent genes expressed in certain cells, tissues, or organs from different organisms and use these "tags" to fish a gene out of a portion of chromosomal DNA by matching base pairs. The challenge associated with identifying genes from genomic sequences varies among organisms and is dependent upon genome size as well as the presence or absence of introns, the intervening DNA sequences interrupting the protein coding sequence of a gene.

An expressed sequence tag or EST is thus a short sub-sequence of a transcribed cDNA sequence. They may be used to identify gene transcripts, and are instrumental in gene discovery and gene sequence determination.[2] The identification of ESTs has proceeded rapidly, with approximately 65,9 million ESTs now available in public databases (e.g. GenBank 18/6/2010, all species). An EST is produced by one-shot sequencing of a cloned mRNA (i.e. sequencing several hundred base pairs from an end of a cDNA clone taken from a cDNA library). The resulting sequence is a relatively low quality fragment whose length is limited by current technology to approximately 500 to 800 nucleotides. Because these clones consist of DNA that is complementary to mRNA, the ESTs represent portions of expressed genes. They may be present in the database as either cDNA/mRNA sequence or as the reverse complement of the mRNA, the template strand. ESTs can be mapped to specific chromosome locations using physical mapping techniques, such as radiation hybrid mapping, Happy mapping, or FISH. Alternatively, if the genome of the organism that originated the EST has been sequenced one can align the EST sequence to that genome using a computer. The current understanding of the human set of genes (as of 2006) includes the existence of thousands of genes based solely on EST evidence. In this respect, ESTs have become a tool to refine the predicted transcripts for those genes, which leads to the prediction of their protein products and ultimately their function. Moreover, the situation in which those ESTs are obtained (tissue, organ, disease state - e.g. cancer) gives information on the conditions in which the corresponding gene is acting. ESTs contain enough information to permit the design of precise probes for DNA microarrays that then can be used to determine the gene expression.

Full genome sequencing (FGS), also known as whole genome sequencing, complete genome sequencing, or entire genome sequencing, laboratory process to determines complete DNA sequence genome at a single time. sequencing all of an organism's chromosomal DNA-including extrachromosomal DNA as well as DNA contained in the mitochondria and for plants the chloroplast as well. Almost any biological sampleeven a very small amount of DNA or ancient DNA can provide the genetic material necessary for full genome sequencing. Such samples may include saliva, epithelial cells, bone marrow, 89

hair (as long as the hair contains a hair follicle), seeds, plant leaves, or anything else that has DNAcontaining cells. Because the sequence data that is produced can be quite large (for example, there are approximately six billion base pairs in each human diploid genome), genomic data is stored electronically and requires a large amount of computing power and storage capacity. Full genome sequencing would have been nearly impossible before the advent of the microprocessor, computers, and the Information Age.

Gene Expression:
The lac Operon in E. coli Introduction
Gene expression is regulated in bacteria, causing genes to be turned on and off in response to environmental signals. You will find information on how genes in bacteria are grouped together in functional groups called operons, the functions of various segments of DNA in operons, how specific molecules start and stop transcription within operons, and how certain molecules affect the rate of this transcription. Animations in the activity show these processes. The DNA of Escherichia coli is sufficient to encode about 4000 proteins, but only a fraction of these are made at any one time. E. coli regulates the expression of many of its genes according to the food sources that are available to it.

Gene Regulation in Bacteria

Bacteria adapt to changes in their surroundings by using regulatory proteins to turn groups of genes on and off in response to various environmental signals. An operon is a cluster of bacterial genes along with an adjacent promoter that controls t he transcription of those genes.

When the genes in an operon are transcribed, a single mRNA is produced for all the genes in that operon. This mRNA is said to be polycistronic because it carries the information for more than one type of protein.

The lac Operator

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The operator is a short region of DNA that lies partially within the promoter and that interacts with a regulatory protein that controls the transcription of the operon.

Here's an analogy. A promoter is like a doorknob, in that the promoters of many operons are similar. An operator is like the keyhole in a doorknob, in that each door is locked by only a specific key, which in this analogy is a specific regulatory protein.

The lac Regulatory Gene

The regulatory gene lacI produces an mRNA that produces a Lac repressor protein, which can bind to the operator of the lac operon.

The general term for the product of a regulatory gene is a regulatory protein. The Lac regulatory protein is called a repressor because it keeps RNA polymerase from transcribing the structural genes. Thus the Lac repressor inhibits transcription of the lac operon.

The Lac Repressor Protein

In the absence of lactose, the Lac repressor binds to the operator and keeps RNA polymerase from transcribing the lac genes.

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Animate
It would be energetically wasteful for E. coli if the lac genes were expressed when lactose was not present. The effect of the Lac repressor on the lac genes is referred to as negative regulation.

The Effect of Lactose on the lac Operon

When lactose is present, the lac genes are expressed because allolactose binds to the Lac repressor protein and keeps it from binding to the lac operator. Allolactose is an isomer of lactose. Small amounts of allolactose are formed when lactose enters E. coli.

Allolactose binds to an allosteric site on the repressor protein causing a conformational change. As a result of this change, the repressor can no longer bind to the operator region and falls off. RNA polymerase can then bind to the promoter and transcribe the lac genes.

The lac Inducer: lactose

lactose is called an inducer because it turns on, or induces the expression of, the lac genes.

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The presence of lactose (and thus allolactose) determines whether or not the Lac repressor is bound to the operator.

Allolactose binds to an allosteric site on the repressor protein causing a conformational change. As a result of this change, the repressor can no longer bind to the operator region and falls off. RNA polymerase can then bind to the promoter and transcribe the lac genes.
Feedback Control of the lac Operon
When the enzymes encoded by the lac operon are produced, they break down lactose and allolactose, eventually releasing the repressor to stop additional synthesis of lac mRNA.

Note
o

Adding a new substrate to the culture medium may induce the formation of new enzymes capable of metabolizing that substrate.

The three enzymes are a permease that transports lactose across the plasma membrane from the culture medium into the interior of the cell beta-galactosidase which converts lactose into the intermediate allolactose and then hydrolyzes this into glucose and galactose. Once in the presence of lactose, the quantity of beta-galactosidase in the cells rises from a tiny amount to almost 2% of the weight of the cell. a transacetylase whose function is still uncertain.

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The capacity to respond to the presence of lactose was always there. The genes for the three induced enzymes are part of the genome of the cell. But until lactose was added to the culture medium, these genes were not expressed (-galactosidase was expressed weakly just enough to convert lactose into allolactose). The most direct way to control the expression of a gene is to regulate its rate of transcription; that is, the rate at which RNA polymerase transcribes the gene into molecules of messenger RNA (mRNA)

DNA Fingerprinting:
Repetitious DNA and DNA polymorphism: In eucaryotes, only a small portion of the genome
constitutes coding regions . In human only about 2% of the genome codes for protein, rest represents noncoding regions. The latter category includes the following types of repetitious DNAs. a) Simple Sequences-a short DNA sequence constitutes one unit and such a unit is repeated tandemly several times.(e.g. Satellite DNA,Microsatellite/Short Tandem Repeats,Minisatellite/Variable Number Tandem Repeat) b) Interspersed Repeats-they are scattered throughout the genome,however they do not form any tandem repeats. DNApolymorphism-It is this noncoding repeat sequences which are the basis for generation of polymorphism among the individuals.The unit length as well as the number of repeats varies from individual to individual,the total length of a specific sequence (say, one VNTR or One STR) at differentclusters will be different in different individuals. The DNApolymorphism involving non-coding simple sequenceDNA, is the genetic basis of DNA fingerprinting. DNAprofiles of no two individuals (except for identicaltwins) are identical The Southern Blot is one way to analyze the genetic patterns which appear in a person's DNA. Performing a Southern Blot involves: 1. Isolating the DNA in question from the rest of the cellular material in the nucleus. This can be done either chemically, by using a detergent to wash the extra material from the DNA,or mechanically, by applying a large amount of pressure in order to "squeeze out" the DNA. 2. Cutting the DNA into several pieces of different sizes. This is done using one or more restriction enzymes. 3. Sorting the DNA pieces by size. The process by which the size separation, "size fractionation," is done is called gel electrophoresis. The DNA is poured into a gel, such as agarose, and an electrical charge is applied to the gel, with the positive charge at the bottom and the negative charge at the top. Because DNA 94

has a slightly negative charge, the pieces of DNA will be attracted towards the bottom of the gel; the smaller pieces, however, will be able to move more quickly and thus further towards the bottom than the larger pieces. The different-sized pieces of DNA will therefore be separated by size, with the smaller pieces towards the bottom and the larger pieces towards the top. 4. Denaturing the DNA, so that all of the DNA is rendered single-stranded. This can be done either by heating or chemically treating the DNA in the gel. 5. Blotting the DNA. The gel with the size-fractionated DNA is applied to a sheet of nitrocellulose paper, and then baked to permanently attach the DNA to the sheet. The Southern Blot is now ready to be analyzed. In order to analyze a Southern Blot, a radioactive genetic probe is used in a hybridization reaction with the DNA in question. If an X-ray is taken of the Southern Blot after a radioactive probe has been allowed to bond with the denatured DNA on the paper, only the areas where the radioactive probe binds [red] will show up on the film. This allows researchers to identify, in a particular person's DNA, the occurrence and frequency of the particular genetic pattern contained in the probe.

1. Hybridization is the coming together, or binding, of two genetic sequences. The binding occurs because of the hydrogen bonds [pink] between base pairs. Between a A base and a T base, there are two hydrogen bonds; between a C base and a G base, there are three hydrogen bonds.

2. When making use of hybridization in the laboratory, DNA must first be denatured, usually by using heat or chemicals. Denaturing is a process by which the hydrogen bonds of the original double-stranded DNA are broken, leaving a single strand of DNA whose bases are available for hydrogen bonding.

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3. Once the DNA has been denatured, a single-stranded radioactive probe [light blue] can be used to see if the denatured DNA contains a sequence similar to that on the probe. The denatured DNA is put into a plastic bag along with the probe and some saline liquid; the bag is then shaken to allow sloshing. If the probe finds a fit, it will bind to the DNA.

4. The fit of the probe to the DNA does not have to be exact. Sequences of varying homology can stick to the DNA even if the fit is poor; the poorer the fit, the fewer the hydrogen bonds between the probe [light blue] and the denatured DNA. The ability of low-homology probes to still bind to DNA can be manipulated through varying the temperature of the hybridization reaction environment, or by varying the amount of salt in the sloshing mixture.

Every strand of DNA has pieces that contain genetic information which informs an organism's development (exons) and pieces that, apparently, supply no relevant genetic information at all (introns). Although the introns may seem useless, it has been found that they contain repeated sequences of base pairs. These sequences, called Variable Number Tandem Repeats (VNTRs), can contain anywhere from twenty to one hundred base pairs. Every human being has some VNTRs. To determine if a person has a particular VNTR, a Southern Blot is performed, and then the Southern Blot is probed, through a hybridization reaction, with a radioactive version of the VNTR in question. The pattern which results from this process is what is often referred to as a DNA fingerprint.

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A given person's VNTRs come from the genetic information donated by his or her parents; he or she could have VNTRs inherited from his or her mother or father, or a combination, but never a VNTR either of his or her parents do not have. Shown below are the VNTR patterns for Mrs. Banerjee [blue], Mr. Banerjee [yellow], and their four children: D1 (the Banerjees' biological daughter), D2 (Mr. Banerjees stepdaughter, child of Mrs. Banerjeeand her former husband [red]), S1 (the Banerjee' biological son), and S2 (the Banerjees' adopted son, not biologically related [his parents are light and dark green]).

Because VNTR patterns are inherited genetically, a given person's VNTR pattern is more or less unique. The more VNTR probes used to analyze a person's

1. Paterniy and Maternity Because a person inherits his or her VNTRs from his or her parents, VNTR patterns can be used to establish paternity and maternity. The patterns are so specific that a parental VNTR pattern can be reconstructed even if only the children's VNTR patterns are known (the more children produced, the more reliable the reconstruction). Parent-child VNTR pattern analysis has been used to solve standard father- identification cases as well as more complicated cases of confirming legal nationality and, in instances of adoption, biological parenthood. 2. Criminal dentification and Forensics DNA isolated from blood, hair, skin cells, or other genetic evidence left at the scene of a crime can be compared, through VNTR patterns, with the DNA of a criminal suspect to determine guilt or innocence. VNTR patterns are also useful in establishing the identity of a homicide victim, either from DNA found as evidence or from the body itself. 3. Personal Identification The notion of using DNA fingerprints as a sort of genetic bar code to identify individuals has been discussed, but this is not likely to happen anytime in the foreseeable future. The technology required to isolate, keep on file, and then analyze millions of very specified VNTR patterns is both expensive and

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impractical. Social security numbers, picture ID, and other more mundane methods are much more likely to remain the prevalent ways to establish personal identification

Chapter 7 : Evolution
Evolution: The process that results inheritable changes in a population spread over many generations (change in allele frequencies over time). Biological evolution refers to populations and not to individuals and that the changes must be passed on to the next generations. Genes mutate, individuals are selected, and populations evolve.

Evidences of evolution

From comparative anatomy

Homologous organs Same basic structural plan and origin but different function It suggests common ancestry Indicates Divergent evolution Thorn of Bougainvillea & Tendril of Cucurbits

Analogous organs Different structure and origin but same function It do not suggests common ancestry Indicates Convergent evolution Thorn of citrus and spine of Opuntia Tendril of cucumbers and tendril of pea

Flipper of seal, wing of bat, cats paw, human hand

Wing of insect and wing of bird

Vestigial organs: Functionless homologous organs that have no apparent function in certain organism.( supposed to be remnants of organs that had been well developed and functional in their ancestors but had became modified during evolution) E.g. 1. Vermiform appendix in man 98

2. Pelvic girdle in python 3. Nictitating membrane 4. Coccyx or tail vertebrae in man

Divergent evolution Origin of a variety of species from a common ancestral form Divergent evolution is the process of two or more related species becoming more and more dissimilar. As they adapted to different environments, the appearance of the two species diverged Homologous organs supports it

Convergent evolution Independent development of similar forms and features by unrelated organisms to adapt to a similar environment Unrelated species become more and more similar in appearance as they adapt to the same kind of environment

Analogous organs supports it

Divergent

Evolution

, the evolutionary pattern in which two species gradually become increasingly different. This type of evolution often occurs when closely related species diversify to new habitats. On a large scale, divergent evolution is responsible for the creation of the current diversity of life on earth from the first living cells.

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On a smaller scale, it is responsible for the evolution of humans and apes from a common primate ancestor. Adaptive radiation is one example of divergent evolution

Convergent Evolution
Convergent evolution takes place when species of different ancestry begin to share analogous traits because of a shared environment or other selection pressure. For example, whales and fish have some similar characteristics since both had to evolve methods of moving through the same medium: water.

Parallel Evolution
Parallel evolution occurs when two species evolve independently of each other, maintaining the same level of similarity. Parallel evolution usually occurs between unrelated species that do not occupy the same or similar niches in a given habitat.

Stabilizing selection favors the norm, the common, average traits in a population In nature, natural selection is most commonly stabilizing. The average members of the population, with intermediate body 100

sizes, have higher fitness than the extremes. Natural selection now acts against change in form, and keeps the population constant through time. Directional selection favours those individuals who have extreme variations in traits within a population. Natural selection may be directional: it may favour, for example, smaller individuals and will, if the character is inherited, produce a decrease in average body size. Directional selection could, of course, also produce an evolutionary increase in body size if larger individuals had higher fitness. Disruptive selection, like directional selection, favours the extreme traits in a population. Disruptive selection differs in that sudden changes in the environment creates a sudden forces favoring that extreme Natural selection can sometimes favour extremes over the intermediate types. This is called disruptive selection. In nature, sexual dimorphism is probably a common example

Directional selection has been studied using artificial selection experiments. Natural selection can also be stabilizing or disruptive. Genetic Drift changes populations.. Random change in allele frequency causes an allele to become common

Founder Effect:
A cause of genetic drift attributable to colonization by a limited number of individuals from a parent population 101

When few individuals colonize a new habitat, genetic drift will more than likely occur.

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The founding population is small and again the alleles present in this small population will not be representative of the original population.

Saltation (from Latin, saltus, "leap") is a sudden change from one generation to the next, that is large, or very large, in comparison with the usual variation of an organism. The term is used for occasionally hypothesized, nongradual changes (especially single-step speciation) that are atypical of, or violate, standard concepts involved in neo-Darwinian evolution. The unorthodox emphasis on saltation as a means of evolutionary change is called saltationism. Gene Flow: Genetic exchange due to the migration of fertile individuals or gametes between populations (reduces differ ences between populations)

Natur al Selecti on: Differ ential success in reproduction; only for m of microevolution that adapts a population to its environment

An example of adaptive radiation these species all diverged from a common ancestor (founder species) 102

FOUNDER SPECIES

Adaptive radiation: The diversification, over evolutionary time, of a species or group of species into several different species or subspecies that are typically adapted to different ecological niches (for example, Darwin's finches). Adaptive radiation is one example of divergent evolution . Artificial selection: The process by which humans breed animals and cultivate crops to ensure that future generations have specific desirable characteristics. (In artificial selection, breeders select the most desirable variants in a plant or animal population and selectively breed them with other desirable individuals). Big bang theory: The theory that states that the universe began in a state of compression to infinite density, and that in one instant all matter and energy began expanding and have continued expanding ever since. Disruptive selection: Selection favoring forms that deviate in either direction from the population average. Selection favours forms that are larger or smaller than average, but works against the average forms between the extremes. Genetic drift: Changes in the frequencies of alleles in a population that occur by chance, rather than because of natural selection.

Gene flow: The movement of genes into or through a population by interbreeding or by migration. Gene frequency: The frequency in the population of a particular gene relative to other genes at its locus, expressed as a proportion (between 0 and 1) or percentage (between 0 and 100 percent). Gene pool: All the genes in a population at a particular time.

Hardy-Weinberg principle: In population genetics, the idea that if a population experienced no selection, no mutation,

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no migration, no genetic drift, and random mating, then the frequency of each allele and the frequencies of genotype in the population would remain the same (constant) from one generation to the next. p2 + 2pq + q2 = 1 (p + q)2 = 1

Calculation of allele frequencies Recessive traits: If the frequency of a recessive trait such as cystic fibrosis or PKU is known, it is possible to calculate allele frequencies and genotype frequencies using the Hardy Weinberg equation and its assumptions as follows:

i. Say 1 in 1, 2500 Indian newborns have cystic fibrosis which means that the frequency of homozygotes for this recessive trait is

q = 1/2,500 = 0.0004

ii. The square root of the frequency of recessives is equal to the allele frequency of the cystic fibrosis allele

q = (0.0004)0.5 = 0.02

iii. The frequency of the normal allele is equal to 1 - the frequency of the cystic fibrosis allele

p = 1- q = 1 - 0.02 = 0.98

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iv. The frequency of carriers (heterozygotes) for the cystic fibrosis allele is

2pq = 2 (0.98)(0.02) = 0.04 or 1/25

v. The frequency of homozygotes for the normal allele is

p = (0.98) = 0.96

vi. Thus the population is composed of three genotypes at the calculated frequencies of

homozygous normal = 0.96 heterozygous carriers = 0.04 homozygous affected = 0.0004

Bottleneck effect- The size of a population becomes drastically reduced by natural disasters ( i.e. volcanic eruptions, earthquakes, fires) will kill individuals non-selectively. The small population left will not be a correct representation of the original population. Some alleles may be over-represented and others under- represented.

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Chronology of Human Evolution: Time period Name Brain capacity Remarks

10-15 Mya

Dryopithecus (ape like)

East Africa Asia ; closely related to Chimpanzee Shivalik hills;Erect posture , small canine teeth 500cc African ape man; height 1.5 meter Tool maker, community life Knew to use fire, larger teeth, East and central asia

Ramapithecus (man like)

2 Mya

Australopithecines ( cave dwellers) Homo habilis

650-800cc

1.5 Mya

Homo eractus (Java ape man)

900cc

100,000-40,000 years ago 25000yrs ago

Neanderthal man

1450cc

Homo sapiens

1650cc

Modern man ; height 1.5 to1.8 meters;f lat face

(Million years ago= Mya)

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Chapter 8 HUMAN HEALTH AND DISEASE

Definition of health
Health is a state of complete physical,mental and social well-being.

Types of diseases
. Infectious easily transmitted from one person to another . .Non- infectious - not transmitted from diseased persons to others.

Common infectious diseases in humans - V iral diseases-common cold, aids,polio,chicken pox,measles - Bacterial diseases -pneumonia,typhoid,cholera - Fungal diseases - ringworms - Helminthic diseases- ascriasis, filariasis Protozoal diseases -malaria, amoebiasis

Name of disease Common cold

Causal organism

Symptoms

Transmission

Rhino virus

Nasal congestion &discharge,sore throat,cough,headache for 3 to 7 days Alveoli get filled with fluid,problem in respiration,fever, chill, cough.

By droplet infection Through Air

Pneumonia

Bacteria- Streptococcus pneumoniae

By inhaling droplets/aerosols released byinfected persons,sharing utensil. By female Anopheles mosquito bite.

Malaria

Protozoa- Plasmodium vivax

Chill, high fever recurring every 3-4 days.due to rupture of RBC.Haemozoin released in blood Chronic inflammation 0f organs,specially lower limbs,genital organs affeected

Filaria

Helminthic worm- Wuchereria bancrofti,W. malayi

By bite of female culex mosquito

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Rringworms

Microsporum,Trichophyton,Epider mo phyton.

Dry scaly lesions on skin,scalp,nail. Intense itching.

From soil,by using clothes, towels,comb of infected person.

LIFE CYCLE OF Plasmodium

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STAGES IN THE LIFE-CYCLE OF Plasmodium

IMMUNITY The overall ability of host to fight with disease causing organism is called immunity. The immune system The system of our body which protects us from various infectious agents and cancer.

IMMUNITY

INNATE

ACQUIRED

Non specific type, present at time of Birth.

An individual acquires it during his life life. Charcteristic features:

Types: a)Physical barrier - skin b)Physiological barrier saliva , tear, acid in stomach. c)Celluar barrier - PMNL neutrophil,macrophages, monocytes.(d) Cytokine - Interferons 1)Specificity 2)Diversity 3)Discrimination between self and non self

Cells involved in acquired immunity Lymphocytes- the major cells of the immune system. They are of 2 types: B lymphocytes ,produced & matured in bone marrow. T lymphocytes, produced at bone marrow but matured in thymus B-cells produce antibodies in response to pathogen in blood. Antibody reacts with antigen(pathogen) & destroys it. 109

This is called humoral immune response as the antibodies are found in the blood Some activated B cells remain as memory cells.

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Structure of an antibody consisting of 2 heavy & 2 light polypeptide chains.

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T-CELLS act differently. They produce a clone of T cells in resonse to pathogen. Helper T cells stimulate B cells & killer T cells to destroy non-self cells. Killer T cells directly attack foreign cells. This type of immune response is known as CELL MEDIATED IMMUNITY (CMI)

TYPES OF ACQUIRED IMMUNITY : ACTIVE IMMUNITY Antibody produced within own body. PASSIVE IMMUNITY Antibodies transferred from another individual.

It is long lasting.

Not long lasting.

Takes time to produce Vaccination and Immunization Vaccination refers to the administration of any vaccine.

Provides immediate relief.

Immunization is the process by which the body produces antibodies in response to the vaccine to fight infections. Vaccine is a preparation of antigenic proteins of pathogens or inactivated/weakened pathogen. It is introduced into the body to generate antibodies which can neutralize the pathogens during actual infection. Vaccines also generate memory B & T cells that recognize the pathogens quickly. Vaccines that contain performed antibodies produce quick immune response and provide Passive Immunity e.g. vaccines against tetanus & snakebite. Other vaccines provide Active Immunity e.g. oral polio vaccine, BCG,cholera vaccine. Allergies The exaggerated response/hypersensitiveness of the immune system of a person to certain antigens coming in contact with or entering into the body is called allergy. 111

Substances that cause allergy are called ALLERGENS.

Common allergens: Dust,pollen,spores,feather,fur,cosmetics, some food substances,heat,cold,sunlight etc. Symptoms of allergy: Sneezing, running nose, watery eyes, difficulty in breathing, itching etc. Drugs to reduce symptoms of allergy: Antihistamines, adrenalin and steroids. Auto-immunity It isa condition when structural & functional damage is caused due to the attack of the self cells of the body by its own immune cells . Examples : Rheumatoid arthritis, Insulin- dependent diabetes. Immune system of the body 1. Primary Lymphoid organs where B &T lymphocytes mature and acquire their antigen specificity e.g.bone marrow and thymus. 2. Secondary lymphoid organs where B & T lymphocytes migrate after maturation and undergo proliferation and differentiation e.g. spleen, lymph nodes, tonsils, Peyers patches of small intestine, appendix. See diagram in page154 of NCERT TEXT BOOK. MALT Mucosal Associated Lymphoid Tissue located within lining of respiratory, digestive and urogenital tracts & forms about 50% lymphoid tissue in human body. AIDS(ACQUIRED IMMUNO DEFICIENCY SYNDROME) Caused by HIV( HUMAN IMMUNO DEFICIENCY VIRUS),a retrovirus. This virus has RNA as its genetic material.

112

Replication of retrovirus

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Transmission of HIV infection i)sexual contact with infected person ii) transfusion of contaminated blood & blood products. iii)by sharing infected needles. iv)from infected mother to her child. Detection a)ELISA(enzyme linked immuno sorbent assay) b)PCR(polymerase chain reaction) c)RIP ( radio immunoprecipitation assay) d)Western blot (cofirmatory test) Prevention i)use of sterile needle, surgical instruments, syringes ii)protected sex iii)counseling before & after the test iv)NACO(National AIDS Control Organisation) has started a number of programmes.

Cancer

Cancer arises from the mutation of a nor ma l gene. Mutated genes that cause cancer are called oncogenes. It is thought that several mutations need to occur to give rise to cancer Cells that are old or not functioning proper ly nor mally self destruct (degenerate) and are replaced by new cells. However, cancerous cells do not self destruct and continue to divide rapidly producing millions of new cancerous cells. Cell division becomes uncontrolled in ca ncer ous cells. 114

Carcinogens

Ionising radiation X Rays, UV light Chemicals Tar from cigarettes Virus infection Papilloma virus can be responsible for cervical cancer. Hereditary predisposition Some fa milies are more susceptible to getting certain cancers. Remember you cant inherit cancer its just that you maybe mor e susceptible to getting it. Benign & malignant tumours Benign tumours do not spread from their site of origin, but can crowd out (squash) surrounding cells eg brain tumour, warts. Malignant tumours can spread from the original site and cause secondary tumours. This is called metastasis.

Cancer detection and diagnosis Detection is based on Biopsy & Histopathological studies of the tissue & blood for increased cell count. In biopsy a thin section of suspected tissue is stained & examined under microscope. Early detection is essential. Radiography, CT, MRI are very useful to detect cancer of internal organs. Cancer Treatment 1-Local therapy: surgery. radiation therapy. 2-Systemic treatment: chemotherapy. Hor monal therapy. Monoclonal antibodies. 3-Supportive care. 115

 4 -Administration of biological response modifier - Interferon given to activate immune system & help in destroying tumour.

Drugs of abuse 1)Opoids : e.g. heroin ,codein, morphene obtained from Papaver somniferum,a depressant 2)Cannabinoids: e.g. hashish,marijuana, charas , ganja. Obtained from Cannabis sativa. effects on cardiovascular system. 3)Coca alkaloid or Cocaine , obtained from Erythroxylum coca.It interferes with transport of neurotransmitter dopamine Causes of drug and alcohol abuse: 1)Curiosity, 2) Need for adventur e, 3)Peer pressure, 4)Excitement, 5) Experimentation, 6) To escape from stress, 7) Unsupportive fa mily structure,8)Repeated use of drugs 7 alcohol. Dependence and addiction to drugs & alcohol leads the patient to (a a) ignor e all social nor ms (,b) social adjustment problems,(c) withdrawl symptoms and (d d) the person may need medical supervision. Warning signs of drug & alcohol abuse These have

1)Drop in academic performance, 2)Absence from school/college, 3)Lack of inter est, 4)Withdrawl, 5)Isolation, 6)Depr ession & fatigue, 7)Aggr essive & rebellious behaviour, 8)Loss of interest in hobbies, 9)Behavioural change, 10)Change in eating & sleeping habits, 11)Stealing, 12)Loss of weight etc.

Prevention and control of drug and alcohol abuse: Avoid undue peer pressure Educating & counselling the child to face problems of life, stresses and to accept failures & disappointments of life Seeking help from parents, teachers,peers & elders to vent their feelings of anxiety,stress & guilt Looking for da nger signals Seeking professional & medical help for deaddiction. Some important definitions:

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Substance abuse It is the use of illicit drugs or the abuse of prescription or over-thecounter drugs for purposes other than those for which they are indicated or in a manner or in quantities other than directed.

Physical dependence It denotes a state when the body of the abuser requires the continuous presence of the drug within it. Psychological dependence When this develops, the substance abuser becomes mentally hooked to the drug & has a continuous uncontrollable craving for it. Some substances that are commonly abused are: Cannabinoids e.g. hashish and marijuana Stimulants e.g.tobacco, nicotine, cocaine, ampheta mines Depr essants e.g. alcohol, barbiturates Narcotics e.g. opioids, morphine derivatives as heroin, opium Hallucinogens e.g. LSD, mescaline Other compounds e.g. steroids, inhalants Probable questions & answers from Human Health and Diseases

Very short answer type question (1 mark)

1. What is disease? Ans. Disease is the condition when the functioning of one or more organ(s) or system(s) of the body is/are adversely affected and characterized by various symptoms. 2. Name one infectious and one non-infectious disease, which are the major causes of mortality? Ans. - Infectious disease - AIDS or Hepatitis-B; - Non-infectious disease Cancer. 3. What are pathogens? Ans. Pathogens are those organisms which cause disease . 4. Name two diseases that spread by droplet infection. Ans. Pneumonia and common cold spread by droplet infection. 5. What name is given to infectious stage of Plasmodium? Ans. Sporozoites. 6. Give the scientific name of pathogen causing malignant malaria in humans. Ans. Plasmodium falciparum. 117

7. What causes swelling of the lower limbs in patients suffering from filariasis? Ans. The filarial worms live on the lymphatic vessels of the lower limbs, where they cause inflammation; that causes swelling of lower limbs. 8. What are interferons? Ans. Interferons are the proteins produced by virus-infected cells in our body. As. 9. Name the two kinds of acquired immunity. Active immunity and passive immunity are the two limbs of acquired immunity.

10. Name the kind of cells responsible for rejecting organ transplants. Ans. T-lymphocytes are responsible for rejecting organ transplants. 1) Why do children of metro cities of India suffer from allergies and asthma? 2 2) A doctor injects preformed antibodies against a snake bite. What type of immunity does it develop in the patient? 1 3) A person has developed allergic reactions like sneezing, watery eyes, running nose and difficulty in breathing. What could be the reason for these symptoms? How can it be controlled? 2 4) A patient has lost his immunity. 3 (i) Name the disease associated with it. (ii) Name the confirmatory test to diagnose the disease. (iii) Why did he lose his immunity? 5) A person claimed that he has seen sounds, heard colours and smelt light. 3 (i) What could be the possible reason? (ii) Name two chemicals responsible for this condition. (iii) Mention any one source for these chemicals.

Short answer type question (2 marks)

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1. Fill in the blanks in the different columns of the table given below : Disease Causative Mode of Symptoms organism transfer Filariasis Wuchereria a Lymphatic vessels of lower limbs affected. b Trichophyton Using towels of infected persons Common cold c Droplets from sneezing of infected persons Ascariasis Ascaris Through contaminated water, vegetables and fruits Dry, scaly lesions on body. Affect nose and Respiratory passage; Sore throat. d

Ans.

(a) Bite of Culex mosquito. (b) Ringworm (c) Rhino virus (d) Blockage of intestinal passage, anemia, muscular pain.

2. How do saliva and tear help to prevent bacterial infection? Ans. -Saliva and tear contain lysozymes. -Lysozymes are the enzymes which digest the cell wall of bacteria. -By lysing the cell wall, they kill the bacteria and prevent their infection. 3. What is vaccination? How does it help producing immunity? 119

Ans.

Vaccination is the process of introducing a preparation of antigenic proteins of the pathogens or weakened or killed pathogen into the body. -The vaccines induce quick multiplication of B and T-lymphocytes; some of them are stored as memory cells. -The B-lymphocytes quickly produce antibodies, which neutralize the antigen during infection.

4. Mention the symptoms of AIDS. Ans. The symptoms of AIDS include: (i) Swollen lymph nodes (ii) night sweats (iii) loss of body weight (iv) fever (v) infection by pathogens of other diseases. 5. Write the full form of ELISA. Give an example of the clinical application of ELISA. Ans. -Enzyme Linked Immuno Sorbent Assay. -ELISA test is used in the diagnosis of AIDS, hepatitis-B and other STDs. 6. List four reasons to justify the ban on intake of cannabinoids by sports persons. Ans. (i) Cannabinoids affect the cardiovascular system. (ii)They cause addiction and when the regular dose is not taken, there are unpleasant withdrawal symptoms, which may be life-threatening. (iii)The person shows reckless behaviour, vandalism and violence. (iv)Excess dose can lead to coma and death due to heart failure, cerebral hemorrhage, etc. Short answer type questions (3 marks) 1. What are the advantages of people being healthy? Ans. When people are healthy, (i) They are efficient at work which consequently increases productivity and brings economic prosperity. (ii) Health increases longevity. (iii) It reduces infant and maternal mortality. 2. Name the two major/broad categories of diseases and differentiate between them. Ans. The two major/broad categories of diseases are: (i) Infectious diseases and (ii) Non-infectious diseases. 120

Differences: Infectious Diseases - These are the diseases which are easily transmitted from one person to another. - They are caused by biological agents, called pathogens. e.g. Diphtheria. Non-infectious Diseases - These are the diseases which are not transmitted from one person to the other. - They occur due to deficiencies, habits, hereditary factors, etc. e.g. Cancer.

3.

(a) Name the respective forms in which the malarial parasite gains entry into (i) Human body and (ii) Body of female Anopheles. (b)Name the hosts where the sexual and the asexual reproductions of malarial parasite occur respectively. (c) Name the toxin responsible for the appearance of symptoms of malaria in humans. Why do these symptoms occur periodically?

Ans.

(a) (i) Sporozoite (ii) Gametocyte (b) - Sexual reproduction in mosquito. - Asexual reproduction in human body. (c) Haemozoin - Haemozoin is released when the RBCs rupture and release the pathogen. - Some cells of pathogen enter fresh RBCs and reproduce asexually and repeat the cycle; hence the symptoms appear periodically. 121

4. Define innate immunity. Name and explain the category of barrier which involves macrophages. Ans. Innate immunity refers to all those defence element with which a person is born and are always available to protect the body. -Macrophanges form part of the cellular barrier. -The cellular barrier includes the following specialized cells; (i) Polymorphonuclear leucocytes. (ii) Monocytes. (iii) Natural killer lymphocytes and (iv) Macrophages. these cells phagocytose and destroy the invading microbes.

5. What is meant by writing H2L2 for an antibody? Name any four types of antibodies produced in our/human body? Ans. - Each antibody molecule has four peptide chains. - Of them, two are small and called light chains (L) and two of them are longer and called heavy chains (H); hence written as H2L2. The four types of antibodies are IgA, IgE, IgG and IgM. 6. Differences between innate and acquired immunity and give examples of each. Ans. Differences: Innate Immunity - Innate immunity consists of all the defence elements with which an individual is born. - It consists of various types of barriers that prevent the entry of foreign agents. - e.g. Lysozymes, mucus-coated epithelium. - It consists of specified cellsB-lymphocytes, T-lymphocytes and antibodies circulating in the body fluids. - e.g. development of immunity by vaccination or by contracting the disease. Acquired Immunity - The immunity which is acquired after birth, is called acquired immunity.

7. Enumerate the steps taken by WHO to control AIDS. 122

Ans.

The steps taken by WHO include: (i) Ensuring use of disposable needles and syringes. (ii) Checking of blood samples for HIV. (iii) Free distribution of condoms. (iv) Advocating safe sex. (v) Controlling drug abuse. (vi) Promoting regular check up for HIV in susceptible cases.

8. How do normal cells get transformed into cancerous neoplastic cells? Mention the differences between viral oncogenes and cellular oncogenes. Ans. The transformation of normal cells into cancerous neoplastic cells is induced by physical, chemical and biological agents collectively called carcinogens. They lose the property of contact inhibition. Difference: Viral Oncogenes - These are the genes present in the oncogenic viruses, which effect oncogenic transformation of the cells they infect. Cellular Oncogenes - These are the genes present in normal cells and code for growth factors; when activated under certain conditions, can cause oncogenic transformation of the cell.

9. What is meant by withdrawal syndrome? What are its characteristic features? Ans. Withdrawal syndrome refers to the characteristic and unpleasant manifestation of the body, when regular dose of drugs/alcohol is abruptly discontinued. Characteristic features: (i) Anxiety (iii) Shakiness (ii) Excess sweating (iv) Nausea

(v) In some cases it may be severe and life-threatening. Long answer type questions (5 marks) 1. What are carcinogens? Describe the different types of them. Ans. Carcinogens are those physical, chemical or biological agents which transform the normal cells into cancerous/neoplastic cells. 123

Types of carcinogens: (i) Physical Agents: - Ionizing radiations like X-rays, gamma rays and - Non-ionizing radiation like UV; these cause damage to DNA leading to neoplastic transformation. (ii) Chemical carcinogens: - Asbestos, cigarette smoke, etc. are carcinogens for lung cancer. (iii) Oncogenic viruses, which possess viral oncogens that transform the normal cells into neoplastic cells. 2. (i) Explain metastasis. Why is it fatal? (ii) The lymphocytes are of two types B and T-cells. Why are they called so? (iii) A person has injured on a road accident and required an urgent immune response. (a) What did the doctor immediately do to this patient? (b) What kind of an immunity was he providing to this patient? (c) Define this type of immunity? (i) Metastasis is the property of tumor cells, which get separated from a tumor, spread to different sites in the body through body fluids and produce secondary tumors wherever they are lodged. Since secondary tumors are formed at several parts of the body, it is difficult to be diagnosed and treated; hence it is fatal. (ii) Those lymphocytes which undergo maturation in the bone marrow are called B-cells while those which undergo maturation in the thymus are called T-cells. (iii) (a) The doctor would inject preformed antibodies. (b) It provides passive immunity. (c) Passive immunity is defined as the immunity developed by directly taking the preformed antibodies that were developed in other vertebrates. 3. (a) Represent schematically the lifecycle of HIV. 124

Ans.

(b) What are the various routes by which transmission of human immunodeficiency virus takes place?

Ans.

(a)

(b) The various routes of transmission of human immunodeficiency virus includes: (i) sexual contact with the infected person. 125

(ii) by transfusion of contaminated blood and blood products. (iii) from infected mother to the child through the placenta. (iv) through contaminated needles and syringes.

Chapter-9 strategies for enhancement in food production

Animal Breeding-objectives:

1.Improved growth rate. 2.Increased production. 3. Improve Desirable Qualities. 4. Improved resistance.

Methods: i).Inbreeding:-Breeding between same breed for 4-6 generations.Ex-cows,baffaloes,poultry In breeding depression- continued in breeding reduces fertility even productivity A single outcross often helps to overcome inbreeding depression ii) Outbreeding- breeding between unrelated animals It is of two types 1. Out crossing- mating within the same breed but not having ancestors.

2.) Crossbreeding- superior males of one breed are mated with superior females of another breed to get better progency.e.g.- cows of inferior breed with superior bull. 3) Hisardale- is a new breed of sheep developed in Punjab by crossing Bikaneri Eves and Marano Rams. 4) Interspecific hybridization- male and female animals of two different species are mated. E.g.- mule is crossbreed of male donkey and female horse. 5) Control breeding- it is done by artificial insemination and multiple ovulation embryo transfer technology (MOET) (a) Artificial insemination- semen of superior male is collected and injected unto the reproductive tract of selected female. The spread of certain diseases can be controlled by this method.

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(b) MOET- This is a technique for herd improvement. Hormones(FSH) are given to the cow for inducing follicular maturation and super ovulation.The cow is either mated with best bull or inseminated .It is done in cattle,sheep,rabbits etc.

Steps in Plant breeding:-1 Collection of variability-Collection and preservation of all different wild varieties,species,relatives of cultivated species etc. are also called germplasm collection. 2.Evaluation and selection of parents-Germplasm is evaluated to identify plants with desirable traits. 3.Cross hybridization among the selected parents-Two plants having two desired characters are hybridized to get new hybrid having two desired characters. 4.Selection and testing of superior recombinants-Selection of the plants having desired character combinations. 5.Testing,release and commercialization of new cultivars-Newly selected lines are evaluated for their yield,agronomic traits,disease resistance etc.and released into the market Green revolution -Crop production. White revolution -Milk production Blue revolution -Fish production Biofortification-Breeding crops with higher levels of vitamins and minerals SCP (Single cell protein )-Microbes such as bacteria,yeast,algae are treated in various ways and used as food. Eg-spirulina can be grown in waste water(from potato processing plant) Tissue culture- cultured part called explant. Types -1.Meristem When apical part is taken and cultured. Uses: a)Rapid clonal multiplication b)Production of virus free plants c)Production of transgenic plants d)Germplasm collection 2. Protoplast culture and somatic hybridization- The plant cell laking cell wall is protoplast. Fusion of protoplast is done by Polyethylene glycol. Pomato is somatic hybrid of potato and tomato.

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Micropropagation-Tissue culture technique used for rapid vegetative propagation of ornamental plants and fruit trees. Somaclone-Plants obtained from single plants by vegetative propagation.

1 Mark questions

Q1.Name two techniques involved in controlled breeding experiments. Ans-a) Artificial insemination(AI) b)Multiple ovulation embryo transfer technology(MOET)

Q2.What is blue and green revolution? Ans- Blue-Increased fish production Green-Crop Q3 What is inbreeding depression? Ans-Loss of vigour due to increase in homozygosity in recessive alleles. Q4. What is heterosis or hybrid vigour? Ans-Phenotypic superiority over either of its parents. Q5.Name the Indian variety of rice patented by an American company. Ans-Basmati. Q6.What is Pomato? Ans-Somatic hybrid of tomato and potato. Q7 Name the algae used as protein rich food. Ans-Spirulina Q8.Expand-MOET and SCP. Ans-MOET-Multiple ovulation embryo transfer technology. SCP-Single cell protein. Q9.What is quarantine? 128

Ans-Careful examination of all introductions for the presence of insects,disease causing organisms. Q10.What is cultivar? Ans-All cultivated varieties of plants. Q1. What is Biofortification? Ans-Breeding crops with higher levels of vitamins and minerals, higher proteins and lesser fats. Objectives of improving-a) Protein content and quality. b)Oil content and quality. c)Vitamins. d)Micronutrient and mineral content.

Q2.Which part of the plant is best suited for making virus free plants? Ans-a) Apical and auxillary meristems b) These are free from virus due to absence of vascular tissues. Q3.What is breed? What are the objectives of animal breeding? Ans- a) Group of animals related by descent,similar in appearance,shape and size b) Improved growth rate , increased production of milk, meat, egg, wool, silk etc. Q4.Define out-crossing? Suggest an advantage. Ans-a) Practice of mating of animals within the same breed,but no common ancestors on either side upto 4-6 generations. b) A single out-cross often helps to overcome inbreeding depression. Q5.What is artificial insemination?what is its importance? Ans_a)Collection of semen from the male bull and injected into the female cow. b)Help to overcome several normal mating problems. Q6. What are the differences between aqua and pisciculture? Ans-a) Production of aquatic plants and animals like fishes , prawns ,lobsters, crabs etc. b) Pisciculture is the production of fishes only. Q7. What is animal husbandry? 129

Ans-Breeding and rearing of livestock like buffaloes ,cows ,pigs, horses , cattle etc.Extended, it includes poultry farming and fisheries also. Q8. What is bird flu? Ans- Contagious disease of animals caused by viruses that only infect birds . symptoms : fever ,cough and sore throat. Q9. Name the most common species of honey bees of India?what are the products from the honey bees? Ans_Apis indica. b) Honey , bee wax and bee venom. Q10. What is germplasm?How it is maintained?

Ans-a)Some total of alleles of genes. b)Low temp.in the form of seeds, Plant tissue culture.

3 Marks Questions Q1.What does inbreeding mean? Suggest its advantages. What is the danger of inbreeding? Ans-a)Mating of more closely related individual within same breed b)i)Development of pure line ii)increases homozygosity iii) Elemination of harmful recessive genes iv)Accumulation of superior genes c) Inbreeding depression-Reduction of fertility and productivity. Q2.Name the methods employed in animal breeding. Which method is the best? Why? Ans-a) Natural methods-these includes inbreeding and out breeding b) Artificial method MOET. It includes super ovulation and embrayo transplantation . Artificial insemination produces good quality progeny , economic . Q3. Explain the procedure of MOET technique in cattle. Ans: FSH is injected to induce follicular maturation and super ovulation . 130

06 to 08 eggs are produced. Cow is inseminated artificially . fertilized eggs transferred to surrogate mother. Q4. What is interspecific hybridization ?Give one example of crop in which it practiced and mention one advantage. Ans : Crossing individuals of two different species to produce a hybrid. Ex- Rice variety ADT 37 from oryza japonica and oryza indica Advantage : Brings useful genetic variations of two or more lines together. Q5. What is cross-breeding ? what advantages does it have? Give example. Ans: Mating between different breeds. Superior male of one and superior female of another breed. Desirable qualities of two breeds are combined Example : Hisardale breed of sheep from Bikaneri ews Merino rams .

5 Marks Questions Q1. Explain the points that have to be considered for successful bee- keeping? Ans : a) Knowledge of nature and habits of bees. b) Selection of suitable location . c) Catching and hiving of swarms. d) Management of bees hives. e) Handling and collection of honey and bee- wax Q2. Write the scientific name of sugarcane grown in north and south India respectively. Mention their characteristic features. Mention the characteristic of the hybrid produced by crossing these two varieties . Ans: (i) North Saccharum barberi poor yield , poor sugar content South Saccharum officinarum thicker stem , higher yield and higher sugar content. (ii) High yield , thick stems , higher sugar content , ability to grow in both North and South India. Q 3. Describe various steps involved in plant breeding . Ans : (i) Collection of variability collection and preservation of all the different wild varieties , species ( germplasm collection )

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ii) Evaluation and selection of parents germplasm is evaluated to identify plants with desirable combination of characters . iii) Cross hybridisation among the selected parents- to bring two good qualities in a hybrid , cross hybridization is done. iv) Selection of testing of superior Recombinants - the superior recombinants are selected and tested . v) Testing , release and comercialisation of new cultivars - newly selected lines are evaluated for yield ,other agronomic traits, disease resistance etc.

Chapter 10: Microbes in Human Welfare

Microbes: Present everywhere. E.g. Thermal vents of geyser (Temp. above 1000c) Deep in soil. Under snow. Diverse. Protozoa, Bacteria, Fungi, Virus, Viroids, Prione (Proteinacious infectious agents) Useful : Antibiotics. Harmful: cause diseases.

In Household Products: Everyday : Lactobacillus (LAB) Lactic acid Bacteria form curd from milk. Increase Vit . B12 Check disease causing microbes in our stomach. Fermentation of dough for dosa, idli (CO2 produced) Making bread bakers yeast. Saccharomyces cerevisiae. Toddy making from sap plam. Cheese making (eg. Swiss chesse by Propionibacterium Sharmanii, Roquefort cheese by fungi.)

In Industrial Products : Beverages and antibiotics. 132

Fermentors : Large vessels for growing microbes.

Fermented Beverages : Beverages like wine, bear, whisky, Brandy, Rum (Saccharomyces cerevisiae) Malted cereals and fruit juices used to produce ethanol, wine and beer produced without distillation; Whisky, brandy, rum produced after distillation.

Antibiotics : (Against life) Penicillin produced by Alexander Fleming from Penicillium notatum while working with Staphylococci Earnest Chain and Howard Plorey awarded Nobel Prize in 1945 for establishing Penicillin as an effective antibiotic. Uses : Treat diseases like plague, whooping cough, diphtheria, leprosy.

Chemicals: Engymes and other Bioactivities Molecules: Uses: Aspergillus niger for production of Citric Acid. A cetobacter aceli for production of Acetic Acid. Clostridium butylicum for production of Butynic Acid. Lactobacillus for production of Lactic acid. Lipases used in detergents to remove oil strains from Laundry. Pectinases and Proteases to clarify bottled jucies. Streprokinase (from Streptococcus) as clot buster in patiento with myocardial infraction (heart attack). Cyclosporin A as inmunosuppresant in organ transplant patients (produced by Trichoderma polysporum) Statins produced by yeast Monascus purpureus used as blood, cholesterol lowering agent.

Microbes in sewage Treatment: Major comporent of waste water, human excreta. Waste water sewage. Cannot be disposed directly into rivers and streams. Before disposal sewage treated in sewage treatment plants (STPs)

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Treatment done in two stages. Primary : Physical removal of particles large and small by filtration and sedimentation. : Solids primary sludge. Supernatant effluent. Secoundry: Primary effluent taken to large aeration tanks. Agitated mechanically and air pumped into it. Aerobic microbes form masses with fungal filaments flocs. Microbes consume organic matter in effluent for growth. BOD ( Biological oxygen demand) reduced. Passed into settling tank. Bacterial flocs sedimented (activated sludge) Small part of activated sludge used as inoculums in aeration tank. Major part pumped into large anaerobic sludge digesters. Anaerobic bacteria digest bacteria and fungi. Bacteria produce gases such as menthane, hydrogen sulphide and CO2 Biogas. Secondary effluent released into rivers and streams. No man made technology available till date. Untreated sewage if released into rivers causes pollution. Ministry of environment and Forests iniated Ganga Action Plan and Yamuna Action Plan.

Biogas plant: Concrete tank 10- 15 mts deep, & slurry or dung fed. Floating cover placed above rises as biogas content rivers. Connecting pipe for supply of biogas. Used for cooking and lighting. Development by IARI :- Indian Agriculture Research institute & KVIC:- Khadi and village Industries Commission.

Microbes as Biocontrol Agents : Insecticides and Pesticides toxic, harmful & are pollutearts. Natural predation better method. No of pests kept in check, not totally eradicated. Food chains not disturbed Eg. Ladybird and Dragon flies useful to get rid of aphids and mosquitoes. Bacillin thureingiensis (Bt) used to control butterfly caterpillar. Mode of spores operation. o Available is sachets, mixed with water and sprayed on plants. o Eaten by insect larva 134

o Toxin released in gut larvae Rpilled. Now Bt toxin genes introduced into plants resistant to insect pests. e.g. Bt cotton. Tungus Trichoderma now being developed. Nucleoply hadrovires good for narrow spectrum insecticide applications. Advantage :No negative impacts on plants, mammals, birds, fish or target insects. For overall IMP (Intergrated pest Management) programme. For ecologically sensitive areas.

AS Biofertilizers: Chemical fertilizers major pollutant. Switch to organic farming and use of biofertilizers need of the time. Main sources of biofertilizers. Bacteria, Fungi & Cyanobacteria. Eg Rhizobium present in roots of leguminious plants fix atmospheric nitrogen into usable organic form. Azospirillium and Azatobacter free living bacteria fix atmosphere Nitrogen. Symbiotic Associations Eg Glomus sp. form myeorrhiza Fungal symbiont absorbs phosphorus from soil and passes it to plant. Plants show resistance to root borne pathogers. Tolerance to salinity and drought Increase in growth and development. Cynobacteria autotrophic fix atmospheric nitrogen Imp biofertilizer. e.g. Anabaena, Nostoc, Oscillatoria. Blue green algae increase fertility by adding organic matter. No of biofertilizers commercially available.

Process of sewage treatment in STP

Primary treatment(physical ) Filtration &sedimentation

Secondary treatment(biological)

Effluent loaded in large aeration tank Agitation & rapid growth of aerobic microbes (flocs)

Consumes organic matter ,reduces BOD

Effluent passed to settling tank

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Flocs sediments form activated sludge

Poured into sludge digester(small amount of a.s.

Very short answer type (1 mark) Name two vitamins produced by microbial fermentation. Vitamin B12 & vitamin B2 What is the botanical name of bakers yeast? Saccharomyces cerevisieae Q. Milk starts to coagulate when lactic acid bacteria is added to warm milk as a starter. Mention two benefits LAB provides? A. i) Besides coagulation, improves nutritional quality by increasing vitaminB12 ii) Check disease causing microbes in our stomach.

Short answer type question(2 marks) State the use of: Trichoderma with respect to organ transplant Nucleo polyhedrovirses with respect to pest management

Ans: Trichoderma polysporum produces a bioactive molecule cyclosporin A Used as an immunosuppresive agents in organ transplant Nucleo polyhedrovirses, a baculovirus attack many insects and arthopods, but do not harm plant, mammals, fish etc. Why should be sewage treated before its disposal? 136

Sewage cantains faecal matter & many pathogenic microbes which would pollute water and cause water borne diseaMention one commercial use of lipase. It is helpful in removing oily stains from the laundry. Statins acts as competetive inhibitor of enzyme responsible for synthesis of cholestrerol What is primary sludge? All the solids that settledown during primary treatment Name the pests, lady birds and dragonflies help to get the rid of respectively Ladybi rds-aphids, dragonflies-mosquitoes. Microbes can be used to lessen the burden of use of chemicals and pesticides-Explin how this can be achieved? Use of friendly microbes which kills & stop the growth. Reduce dependency of chemicals of bacteria- Bacillus thuringenesis, used to kill bollworm catterpillars on cotton Give the role of microbes in single cell protein. SCP is protein rich microbial biomass which can be used as food and feed. SCP has all essential amino acid, fat content is low. Spirulina has become an important food supplement What is micorhiza? How does it help as biofertilizers? Association between roots of higher plants and fungus Solubilize phosphorus and produce growth promoting substances It protects plants from soil pathogens What is BOD? What does it mean if water samples has more BOD? Biochemical oxygen demands. It represent the amount of dissolved oxygen that would be consumed if all the organic matter in one litter of water is oxidized by microbes More value of BOD means the water sample is polluted by organic matter. Name any two cyanobacteria. How do they serve as main source biofertizer. Nostoc & anabaena. They possess heterocyst and fix atmospheric nitrogen into ammonia. Fixed form of nitrogen leaches out in soil & is absorbed by higher plants What is the difference between Bt and Bt cotton? Explain the use of Bt as a biological control.

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Bt stands for bacteria name Bacillus thuringenesis whose gene has been incorporated in cotton plant to make Bt cotton. Bt is mixed with water & sprayed, It is eaten by insect larvae, in the gut toxin is released and insect is killed Name the blank spaces a, b, c, d in the table given belowTypes of microbes Bacterium b c Name a Aspergillus niger d Commercial product Clot buster enzyme Citric acid Butyric acid

a, Steptococcus: b, Fungus: c, Bacterium: d, Clostridium butylicum

Give reason- a) Bottled fruit juices brought from market are clearer as compared to those made at home, b)Large holes are found in swiss-cheese, c) The insect which are so called pest are not eradicated in organic fumes. a) Clarified by the use of pectinases and proteses, b) large holes due to production of large amount of carbon di oxide by bacterium Propionibacterium sharmanii, c) insect pest are not eradicated because the beneficial predatory insect and other parasites which depend upon them as food or host would not be able to survive

Name the gobar gas liberated from biogas plant. which type of bacteria are responsible for its production? Give advantage. It is methane gas. By the activity of methanogenic bacteria

Advantage: it is readily combustible, it do not pollute environment

Q. Short answer type question(3 marks) Differentiate between a) primary sludge and activated sludge, b) biofertilizer and chemical fertilizer, c) primary sewage treatment and secondary sewage treatement.

Primary sludge
All solids that settle down during primary treatment Biofertilizer There are micro-organism which enrich their nutrient quality of the soil

Activated sludge
The sediment formed during secondary treatment Chemical fertilizer There are the chemicals synthesized in 138 factories and give only specific nutrient

Primary sewage treatment It involves the physical removal of particles by filtration and sedimentation

Secondary sewage treatment Pretreated effluent is biologically treated with microbes in aeration tank.

Q. Draw labelled diagram of a bacteriophage

Q. Label the parts numbering 5 ,3 ,9 , 1 of a typical biogas plant.

Give the role of spent slurry. Constituent of biogas 5) floating gas holder, 3) sludge, 9) digester, 1) Dung and water

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Spent slurry is used as fertilizer, Constituent of biogas is CH4+CO2+H2

Long answer type question(5 marks) Answer briefly: 1) How is sewage harmful to men? 2) What is organic farming? 3)which group of organisms attack insect and arthopod? How are they best biocontrol biological agent, 4) What is the difference between flocks and primary sludge?

Sewage is municipal waste water which contains human excreta, other waste and pathogenic microbes The natural method of pest and pathogen control involving use of viruses, bacteria and other insect i.e. which are the natural predator and pest Baculovirus : species specific ,narrow- spectrum ,eco-friendly insecticides. Primary-Sludge sands, silt, small pebbles settledown during primary treatment. fungal filament which like structure.

Flocks masses of bacteria with form mesh

Write short notes on: a) bakers yeast, b) alcohol c) statin d)Brewers yeast e) streptokinase

Probable Questions. MICROBES IN HUMAN WELFARE

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Q1 what are biofertilizers ? give 1 example. Q.2 Name the bacteria used in the production of "Swiss Cheese" Q.3 What is the scientific name for Brewer's yeast? Q.4 Name a bioactive molecule used as an immuno suppressive agent. Q.5 Why are bottled fruit juices appear clearer than the home made ones. Q.6 Three water samples labelled A, B & C has BOD values 20mg/L, 8mg/L and 400 mg/L respectively. Which sample of water is most polluted and which one most clean. Hint : The greater the BOD of waste water, more is its polluting potential. Q7) Name the group of bacteria that are capable to live in high temperatures above 100 C Q8) To which class of fungi does Penicillium belong? (1) Q9_ Which of the steps are related to Nitrogen fixation? (1) a) N2 NH3 b) N2 NO3 c) N2 Amino acid d) Both a and b Q10) Which of the following organisms has Nif Gene? (1) a) Penicillum b) Rhizobium c) Aspergillus . d) Streptococcus 2 Marks Questions Q.1 What are statins? What is its significance? Q.2 In which food will you find LAB? Mention its two useful applications. Q.3 can microbes be used to decrease the use of chemical fertilizers and pesticides? Explain how? What is BOD? What does it indicates .

3 Marks Questions Q.1 Draw a labelled diagram of a typical biogas plant. Q.2 Explain Why : (a) Cow dung is used in the generation of biogas. (b) A small amount of curd is added in fresh milk to convert it into curd. (c) Baculovirus are used in narrow spectrum insecticidal application. Q.3 What are antibiotics? Give two examples. What is their significance? Give one example and one use of the following (3) a) Free living fungi b) Symbiotic fungi c) Free living bacteria Q4) How is it that the Cry protein produced by Bacillus thuringiensis (Bt) does harm the bacteria but only killsthe insect larvae?

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Q5) Differentiate Antibodies and antibiotics Q6) How are biofertilizers different from fertilizers such as NPK that we buy in the market? Justify the role of Rhizobium as a biofertilizer. 5 Marks Questions Q1)Explain how microbes are used in sewage treatment? Q2)What do you understand by integrated pest management (IPM)? Explain with example and state its importance

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Chapter-11: BIOTECHNOLOGY: PRINCIPLES AND PROCESSES

Biotechnology is a broad area of science involving multiple disciplines designed to use living organisms or their products to perform valuable industrial or manufacturing processes or applications pertaining to human benefit.

Recombinant DNA technology: An organism's genome contains virtually all the information necessary for its growth and development Determining the molecular sequence of DNA that makes up the genome of different organisms is an international scientific goal, several laboratories are participating worldwide in this task. It is thought that having access to the complete DNA sequence of an organism can help us not only to decipher its biology but also help us understand major biological questions, for instance, what makes one species pathogenic whereas a related species is not. Each cell (with a few exceptions) carries a copy of the DNA sequences which make up the organism's genome. However, many genomes are large and complex (for instance the human genome is made up of ~3000 x 106 base pairs). A particular DNA sequence (for instance the allele of a gene) can be very small in comparison. And it probably occurs only once or twice within the genome (ie only one or two copies per cell). This means that a particular DNA sequence will be present as only a (very) small part within the complex mixture of DNA sequences that make up the genomic DNA of that organism. It is often necessary to 'break up' large DNA molecules into smaller, more manageable fragments - often to sizes ranging from 100 bp to 2 kb (bear in mind that each resulting DNA fragment is an individual molecule). These smaller fragments can then be manipulated more easily - to isolate particular DNA fragments, to characterise their molecular sequence, to determine their function, to determine their position in relation to other sequences within the genome, to use them to express proteins, etc. . Progress in understanding genetic mechanisms at the molecular level was slow. Then came the discovery of various bacterial and viral enzymes which modify and synthesise nucleic acids (DNA and RNA), along with the means to produce more out with the organism from which they were originally isolated. The application of these enzymes for manipulating DNA (no matter what the source) led to the creation of Recombinant DNA Technology which has enabled great scientific advances in the field of biology, has created new scientific disciplines and has revolutionised our world. Recombinant DNA Technology Techniques for - Isolation - Digestion - Fractionation - Purification of the TARGET fragment - Cloning into vectors - Transformation of host cell and selection

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- Replication - Analysis - Expression of DNA

Restriction enzymes . Restriction enzymes are endonucleases Bacterial enzymes Different bacterial strains express different restriction enzymes The names of restriction enzymes are derived from the name of the bacterial strain they are isolated from Cut (hydrolyse) DNA into defined and reproducible fragments Basic tools of gene cloning Names of restriction endonucleases Titles of restriction enzymes are derived from the first letter of the genus + the first two letters of the species of organism from which they were isolated. EcoRI - from Escherichia coli BamHI - from Bacillus amyloliquefaciens HindIII - from Haemophilus influenzae PstI - from Providencia stuartii Sau3AI - from Staphylococcus aureus AvaI - from Anabaena variabilis Enzymes that can cut (hydrolyse) DNA duplex at specific sites. Current DNA technology is totally dependent on restriction enzymesThe EcoRI restriction enzyme--the first restriction enzyme isolated from E. Coli bacteria--is able to recognize the base sequence 5' GAATTC 3'. Restriction enzymes cut each strand of DNA between the G and the A in this sequence. This leaves "sticky ends" or single stranded overhangs of DNA. Each single stranded overhang has the sequence 5" AATT 3'. These overhanging ends will bond to a fragment of DNA which has the complementary sequence of bases

Restriction enzymes recognise a specific short nucleotide sequence

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This is known as a Restriction Site The phosphodiester bond is cleaved between specific bases, one on each DNA strand

The product of each reaction is two double stranded DNA fragments Restriction enzymes do not discriminate between DNA from different organisms Most restriction enzymes will cut DNA which contains their recognition sequence, no matter the source of the DNA Restriction endonucleases are a natural part of the bacterial defence system Part of the restriction/modification system found in many bacteria These enzymes RESTRICT the ability of foreign DNA (such as bacteriophage DNA) to infect/invade the host bacterial cell by cutting it up (degrading it) The host DNA is MODIFIED by METHYLATION of the sequences these enzymes recognise o Methyl groups are added to C or A nucleotides in order to protect the bacterial host DNA from degradation by its own enzymes

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Hundreds of restriction enzymes have been isolated and characterised Enables DNA to be cut into discrete, manageable fragments Type II enzymes(II Recognise a specific target sequence in DNA, and then break the DNA (both strands), within or close to, the recognition site) e.g. EcoRI, these are those used in the vast majority of molecular biology techniques .

Many Type II restriction endonucleases recognise PALINDROMIC sequences (From Greek palindromos, running back again, recurring : palin, again) A segment of double-stranded DNA in which the nucleotide sequence of one strand reads same in reverse order to that of the complementary strand. (always read from the same direction)

For example, EcoRI recognises the sequence 5'-G A A T T C-3' 3'-C T T A A G-5'

Different enzymes cut at different positions and can create single stranded ends ('sticky ends') with overhangs Some generate 5' overhangs - eg: EcoRI

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Some generate 3' overhangs - eg: PstI

Some generate blunt ends- eg: SmaI

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The 'sticky' overhangs are known as COHESIVE ENDS The single stranded termini (or ends) can base pair (ANNEAL) with any complementary single stranded termini This is the basis for RECOMBINANT DNA TECHNOLOGY Inserting foreign DNA into a cloning vector Restriction enzymes are a useful tool for analysing Recombinant DNA After ligating a particular DNA sequence into a cloning vector, it is necessary to check that the correct fragment has been taken up. Sometimes it is also necessary to ensure that the foreign DNA sequence is in a certain orientation relative to sequences present in the cloning vector. Checking the size of the insert Checking the orientation of the insert Determining pattern of restriction sites within insert DNA DNA fractionation Separation of DNA fragments in order to isolate and analyse DNA cut by restriction enzymes Electrophoresis Electrophoresis is a technique used to separate and sometimes purify macromolecules especially proteins and nucleic acids - that differ in size, charge or conformation. As such, it 148

is one of the most widely-used techniques in biochemistry and molecular biology Linear DNA fragments of different sizes are resolved according to their size through gels made of polymeric materials such as polyacrylamide and agarose. For instance, agarose is a polysaccharide derived from seaweed - and gels formed from between 0.5% to 2% (mass/volume i.e. 0.5 to 2.0g agarose/100 ml of aqueous buffer) can be used to separate (resolve) most sizes of DNA When charged molecules are placed in an electric field, they migrate toward either the positive or negative pole according to their charge. In contrast to proteins, which can have either a net positive or net negative charge, nucleic acids have a consistent negative charge imparted by their phosphate backbone, and migrate toward the anode DNA is electrophoresed through the agarose gel from the cathode (negative) to the anode (positive) when a voltage is applied, due to the net negative charge carried on DNA

When the DNA has been electrophoresed, the gel is stained in a solution containing the chemical ethidium bromide. Ethidium bromide is a fluorescent dye that intercalates between bases of nucleic acids and allows very convenient detection of DNA fragments in gels, This compound binds tightly to DNA (DNA chelator) and fluoresces strongly under UV light - allowing the visualisation and detection of the DNA. Like any molecule that binds to DNA, Ethidium bromide is hazardous. It is a carcinogen. Finally,, the separated DNAs are cut out from the gel and extracted from the gel piece(elution).

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Fig:Seperation of DNA fragments according to size. Recombinant DNA technology: DNA cloning is the isolation of a fragment or fragments of DNA from an organism and placing in a VECTOR that replicates independent of chromosomal DNA. The RECOMBINANT DNA is propagated in a host organism, the resulting CLONES are a set of genetically identical organisms which contain the recombinant DNA Why is DNA cloning important? DNA cloning is involved in a number of applications (GENETIC ENGINEERING). Many techniques for DNA isolation and manipulation have been worked out and are now routinely followed in scientific laboratories. Three main purposes for cloning DNA 1) DNA sequencing Determining the sequence of the bases in the DNA can tell us about which proteins or RNAs are encoded and their sequences, which sequences control their expression (GENE PROMOTERS and other control sequences), as well as any possible mutations which might alter their function.

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Having access to the complete DNA sequence of an organism can help us decipher the biology of that organism. 2) Protein production Isolating the gene which encodes a desired protein (haemoglobin, interferon) may allow that gene to be over-expressed so that the protein can be produced in bulk for study or use 3) Engineering animals/plants/proteins The ability to alter the properties of proteins as well as create genetically modified animals and plants (TRANSGENICS) has lead to their use for research and for therapeutic and commercial purposes. The technology may lead to the development of new therapies for the treatment of disease (GENE THERAPY). Cloning and Expression Vectors Isolated DNA is cloned into VECTORS for long term storage, propagation of the DNA and for production of protein from gene(s) encoded in the DNA What are cloning vectors? Cloning vectors are extra-chromosomal 'replicons' of DNA which can be isolated and can replicate independently of the chromosome. Vectors usually contain a selectable marker - a gene that allows selection of cells carrying the vector e.g. by conferring resistance to a toxin. DNA of interest can be cloned into the vector and replicated in host cells, usually one which has been well characterised. Commonly used vector systems Bacterial plasmids Bacteriophages Cosmids Yeast artificial chromosomes (YACs) Ti plasmid (plants) Eukaryotic viruses such as baculovirus (insect cells), SV40 virus and retroviruses. Characteristics of a Cloning Vector
Origin of replication (ORI)

This process marks autonomous replication in vector. ORI is a specific sequence of nucleotide in DNA from where replication starts. When foreign DNA is linked to this sequence then along with vector replication, foreign (desirable) DNA also starts replicating within host cell.
Selectable Marker

Besides ORI, a cloning vector should have selectable marker gene. This gene permits the selection of host cells which bear recombinant DNA (called transformants) from those which do not bear rDNA (nontransformants). Charecteristics of Selectable marker:

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A gene whose expression allows one to identify cells that have been transforrned or transfected with a vector containing the marker gene. A marker gene is used to determine if a piece of DNA has been successfully inserted into the host organism.. A gene, usually encoding resistance to an antibiotic, added to a vector construct to allow easy selection of cells that contain the construct from the large majority of cells that do not. Gene that allows for selection of cells carrying the gene. More commonly, selectable markers provide resistance against cytotoxic insults .A selectable marker will protect the organism from a selective agent that would normally kill it or prevent its growth. In most applications, only one in a several billion cells will take up DNA. Rather than checking every single cell, scientists use a selective agent to kill all cells that do not contain the foreign DNA, leaving only the desired ones .Antibiotics are the most common selective agents. In bacteria, antibiotics are used almost exclusively. Restriction sites It should have restriction sites, to allow cleavage of specific sequence by specific Restriction Endonuclease. Restriction sites in E.coli cloning vector pBR322 include HindIII , EcoRI , BamHI , SalI, PvuI, PstI, ClaI etc.

A Cloning Vector that Works with Plant Cells The most commonly used plant cloning vector is the "Ti" plasmid, or tumor-inducing plasmid. This plasmid is found in cells of the bacterium known as Agrobacterium tumefaciens, which normally lives in soil. The bacterium has the ability to infect plants and cause a crown gall, or tumorous lump, to form at the site of infection. The tumor-inducing capacity of this bacterium results from the presence of the Ti plasmid. The Ti plasmid itself, a large, circular, double-stranded DNA molecule, can replicate independently of the A. tumefaciens genome. When these bacteria infect a plant cell, a 30,000 base-pair segment of the Ti plasmid - called T DNA - separates from the plasmid and incorporates into the host cell genome. This aspect of Ti plasmid function has made it useful as a plant cloning vector (natural geneticengineer). The Ti plasmid can be used to shuttle exogenous genes into host plant cells. This type of gene transfer requires two steps: 1) the endogenous, tumor-causing genes of the T DNA must be inactivated and, 2) foreign genes must be inserted into the same region of the Ti plasmid. The resulting recombinant plasmid, carrying up to approximately 40,000 base pairs of inserted DNA and including the appropriate plant

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regulatory sequences, can then be placed back into the A. tumefaciens cell. That cell can be introduced into plant cell protoplasts either by the process of infection or by direct insertion. Once in the protoplast, the foreign DNA, consisting of both T DNA and the inserted gene, incorporates into the host plant genome. The engineered protoplast - containing the recombinant T DNA - regenerates into a whole plant, each cell of which contains the inserted gene. Once a plant incorporates the T DNA with its inserted gene, it passes it on to future generations of the plant with a normal pattern of Mendelian inheritance.

Plasmids are the most commonly used vector system. Several types are available for cloning of foreign DNA in the host organism Escherichia coli. Many E. coli plasmids allow the expression of proteins encoded by the cloned DNA

Bacteriophage are another common vector system used for cloning DNA. These are viruses which 'infect' E. coli. The M13 bacteriophage is a single-stranded DNA virus which replicates in E. coli in a double-stranded form that can be manipulated like a plasmid. It can be used to produce single-stranded DNA copies which are useful for DNA sequencing. Strategy for cloning DNA into a plasmid (or other cloning) vector SUBCLONING cut DNA of interest with the appropriate restriction endonuclease(s) separate fragments by gel electrophoresis purify target fragment from gel ligate fragment with a plasmid cut with the same restriction endonuclease(s) - ligation is performed using the enzyme DNAligase, ATP and Mg2+ ions Transformation is the process by which plasmids (or other DNA) can be introduced into a cell. For E. coli transformation with plasmids is quite straightforward, plasmids can be introduced by electroporation or by incubation in the presence of divalent cations (usually Ca2+) and a brief heat shock (42C) which induces the E. coli cells to take up the foreign DNA There are different methods to select for transformed cells. For instance, transformants can be selected as antibiotic-resistant colonies on agar plates containing antibiotic. For E. coli transformed with plasmids, colonies grown on antibiotic-containing plates should all carry plasmids. However, this does not guarantee that the plasmid contains an insert. It is possible that the vector has re-ligated and not incorporated an insert. A means to determine which clones contain plasmids with inserts is to use a positive control method such as insertional inactivation. This provides a clear way of recognising recombinants from those carrying re-ligated vector. The following two methods are most commonly used 1. antibiotic selection and replica plating 153

2. colour selection: blue/white selection using the lacz gene Insertional inactivation Subcloning a DNA fragment into an active gene (usually a marker gene whose function can be easily detected) will disrupt the function of that gene. This can be detected by looking for colonies that no longer display that phenotype. Colour selection A more common method to determine which transformants contain plasmids with inserts is to use colour selection. For E. coli, this involves the lac complex and blue/white screening. The lac complex is comprised of the LacI gene and the lacZ gene. The gene lacZ produces the enzyme beta-galactosidase, which can break down a derivative of lactose called X-gal (5-bromo-4-chloro-3indolyl-beta-D-galactopyranoside). This produces a blue dye, which colours the colony blue. In E. coli, the expression of the lacZ gene is regulated by the LacI repressor protein (product of the LacI gene) which binds to the promoter, and prevents transcription until another lactose analogue is added. IPTG (isopropyl-beta-D-thiogalactopyranoside) is a non-degradable lactose analogue which binds to the LacI repressor protein and displaces it from DNA it binds in the lac promoter system. Transcription of lacZ can then occur and X-gal can be broken down. Usually, modern cloning vectors encode a shortened derivative of lacZ (lacZ'), which encodes the Nterminal alpha-peptide of beta-galactosidase. A strain of E. coli is used which produces only the Cterminal portion of the enzyme, the transformant can use the reconstituted enzyme to break down X-gal. The polylinker site in these vectors is located within the lacZ' gene.

Cloning an insert into this site will disrupt the gene and recombinants will be unable to break down X-gal. Colonies carrying plasmid with no insert will be coloured blue whereas colonies carrying recombinant plasmid will be white.

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For plasmids such as pBR322, which contains two antibiotic resistance genes, cloning an insert into one of these will disrupt that gene and inactivate the resistance to that antibiotic.

For example, if DNA is cloned into the BamHI site in the Tetr gene, then transformed bacteria should be plate onto media containing the other antibiotic (ampicillin). Positive clones can then be selected by replica plating.

Colonies are transferred by replica plating on to identical positions on two more plates, each of which contains one of the antibiotics which the parental plasmid confers resistance to - in this case tetracycline and ampicillin. Transformants which carry plasmids with insert should be unable to grow on the tetracycline plate and can be selected from the identical position on the ampicillin plate on which they should still be able to grow. Analysis of clones One of the first steps is to identify clones carrying the recombinant plasmid, with the desired DNA insert. This can be done by 'picking' clones - choosing individual bacterial colonies in order to isolate the plasmid DNA from each of them. Single bacterial colonies are grown in culture broth containing the selection antibiotic in order to maintain the plasmid. The plasmid DNA is 155

extracted by the standard minipreparation technique and then analysed by restriction digest. After digesting the DNA, different sized fragments are separated by agarose gel electrophoresis and the sizes determined by comparison with known DNA molecular weight markers. 1. PCR(Polymerase Chain Reaction): The polymerase chain reaction (or PCR) is a technique for the in vitro amplification of a desired sequence of DNA. PCR allows the generation of a large quantity of DNA product (up to several g) from only a few starting copies. It has been shown that PCR can be used to generate a detectable quantity of DNA from only one starting target (or template) molecule. PCR was developed in the mid-1980's, but has already found multiple applications, such as: 1. Rapid amplification of intact genes or gene fragments 2. Generation of large amounts of DNA for sequencing 3. Generation of probes specific for uncloned genes by selective amplification of a specific segment of cDNA 4. Analysis of mutations for medical applications 5. Detection of minute amounts of DNA for forensic purposes 6. Amplification of chromosomal regions adjacent to genes of known sequence and many more Development of PCR won the Nobel prize for Kary Mullis and co-workers. PCR principle The PCR reaction is a DNA synthesis reaction that depends on the extension of primers annealed to opposite strands of a dsDNA template that has been denatured ( melted apart) at temperatures near boiling. By repeating the melting, annealing and extension steps, several copies of the original template DNA can be generated. The amount of starting material (target) needed is very small It is not necessary to isolate the desired sequence, because it will be defined by the primers that are used in the reaction. The primers are oligonucleotides complementary to different regions on the 2 strands of DNA template (flanking the region to be amplified). The primer acts as a starting point for DNA synthesis. The oligo is extended from its 3' end by DNA polymerase.

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Primer design PCR is a cycle of three steps: 1. DENATURATION - the strands of the DNA are melted apart by heating to 95C 2. ANNEALING - the temperature is reduced to ~ 55C to allow the primers to anneal to the target DNA 3. POLYMERISATION/EXTENSION - the temperature is changed to the optimum temperature in order for the DNA polymerase to catalyse extension of the primers, i.e. to copy the DNA between the primers. The cycle is repeated over and over again - as many times as needed to produce a detectable amount of product. Typical cycling regime Approximately 20-40 cycles of the 3 reaction steps are performed in a PCR reaction. A typical reaction sequence would be: Initial denaturation - 95C for 2 mins 30 cycles of 95C (30 seconds denaturation) 158

55C (30 seconds primer annealing) 72C (60 seconds primer extension, for fragments up to 1 kb) Final extension of all DNA ends - 72C for 10 mins Storage of DNA - 4C

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Discovery of a thermostable DNA polymerase The breakthrough came with the discovery of the thermostable DNA polymerase Taq polymerase, from the thermophilic bacterium, Thermus aquaticus, which lives in hot springs., Taq polymerase enzyme can resist the high temperatures required to melt the template DNA apart without denaturation (loss of activity) and works best at high temperatures (72C). This led to improved specificity & sensitivity. Annealing of primers to sites other than the target sequence is significantly reduced at the higher temperatures used for Taq polymerase. The fact that only one aliquot ot Taq polymerase needs to be added to the PCR reaction means that all components can be added to the tube at the start of the reaction - allowing the automation of the PCR. Heating blocks which can be programmed to carry out the time and temperature cycles for a PCR have been developed (thermal cycler machine).

Applications of PCR
1) Cloning a gene encoding a known protein 2) Amplifying 'old DNA' 3) Amplifying cloned DNA from vectors Convenient way of checking the inserts - amplified DNA can be analysed by electrophoresis, Southern blotting 4) Creating mutations in cloned genes 5) Rapid amplification of cDNA ends - RACE . 6) Detecting bacterial or viral infection * AIDs infection * Tuberculosis (Mycobacterium tuberculosis) 7) Cancer Detecting mutations that occur in cancer and monitoring cancer therapy. Determining if a patient is free of malignant cells 160

8) Genetic diagnosis a. Diagnosing inherited disorders * Cystic fibrosis * Muscular dystrophy * Haemophilia A and B * Sickle cell anaemia b. Diagnosing cancer - certain cancers are caused by specific and reproducible mutations: e,g. Retinoblastoma - childhood cancer of the eye. The heritable form (germ line mutation of one of the two retinoblastoma allelles): mutation is detected in all cells. Spontaneous form: only detected in tumour tissue. c. Blood group typing d. Prenatal diagnosis - such as determining the sex of foetuses for those at risk of X-linked disorders PCR is one of the most versatile techniques invented, and has so many applications that this list could go on for quite some time.

Downstream processing
It refers to the recovery and purification of biosynthetic products, particularly pharmaceuticals, from natural sources such as animal or plant tissue or fermentation broth, including the recycling of salvageable components and the proper treatment and disposal of waste. It is an essential step in the manufacture of pharmaceuticals such as antibiotics, hormones (e.g. insulin and human growth hormone), antibodies (e.g. infliximab and abciximab) and vaccines; antibodies and enzymes used in diagnostics; industrial enzymes; and natural fragrance and flavor compounds. Downstream processing is usually considered a specialized field in biochemical engineering, itself a specialization within chemical engineering, though many of the key technologies were developed by chemists and biologists for laboratory-scale separation of biological products.

Stages in Downstream Processing

A widely recognized heuristic for categorizing downstream processing operations divides them into four groups which are applied in order to bring a product from its natural state as a component of a tissue, cell or fermentation broth through progressive improvements in purity and concentration. Removal of insolubles Product Isolation ProductPurification Product Polishing

A) 1 Mark Questions
1) 2) 3) 4) 5) 6) What is biotechnology? Define plasmid. What are molecular scissors? What do you mean by recognition sequence? Which enzymes act as molecular glue? What is elution? 161

7) What are cloning vectors? 8) Name the sequence within a cloning vector from where the replication commence. 9) Mention the bacteria that acts as natural genetic engineer. 10) Name any two processes by which alien DNA is introduced into the host cell. 11) Expand the term PCR. 12) Name the microorganism from which the thermostable DNA polymerase required for PCR is obtained? 13) What is a bioreactor? 14) What are the two main processes involved in downstream processing? HINTS: 1)Large scale production and marketing of products and processes usin g living organisms,cells or enzymes. 2)Autonomously replicating circular , extra-chromosomal bacterial DNA used in gene manipulation. 3)Restriction enzymes. 4) Restriction endonucleases always cut DNA at a specific point by recognizing a specific sequences of base pair known as recognition sequence. 5) DNA ligases 6)The ultimate step in the separation and isolation of DNA fragments through gel electrophoresis in which separated bands of DNASs are cut out from the gel and extracted from the gel piece. 7) Cloning vectors are extra-chromosomal 'replicons' of DNA which can be isolated and can replicate independently of the chromosome.. DNA of interest can be cloned into the vector and replicated in host cells 8)ORI point 9) Agrobacterium tumefaciens 10)Micro injection,biolistics(gene gun) 11)Polymerase Chain Reaction 12) Thermus aquaticus 13)Large scale biotechnological product involves the use of bioreactor. 14) Separation and purification.

2-Marks Questions
1)Enlist the core techniques that pave the way for modern biotechnology.

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2) What is gene cloning? 3)Mention the three steps involve in genetically modifying an organism. 4) Why do bacteria possesses restriction enzyme ? 5)Mention one basic difference between restriction endonucleases and exonucleases. 6) What is a palindromic sequence? Give example. 7) What are sticky ends and blind ends ? 8)Mention the role of selectable marker in cloning vector. 9)What is insertional inactivation? 10) Howe can you make a bacterial cell competent to take up foreign DNA ? HINTS: 1) ( a) Genetic engineering (b) maintenance of sterile ambience. 2) The process of cloning multiple copies of a gene. 3) (a) identification of DNA with desirable genes (b) introduction of the identified DNA into the host and (c) maintenance of introduced DNA in the host and transfer of DNA to its progeny. 4) By restriction enzyme bacteria can attack and destroy the phage DNA in a case of viral attack and thereby prevent viral attack. 5) Exonucleases digest DNA from the flank ( beginning/end) of the DNA strands. Whereas endonucleases catalyses the hydrolytic cleavage of DNA in the middle. 6) A segment of double-stranded DNA in which the nucleotide sequence of one strand reads same in reverse order to that of the complementary strand. (always read from the same direction) 7) Double stranded ends of a DNA molecule (without any overhangings) produced by the action of certain restriction enzymes .-[blunt ends)/ Sticky ends - Double stranded ends of a DNA 0molecule (with overhangings) produced by the action of certain restriction enzymes 8) The selectable marker genes in a cloning vector allow fopr the selection and identification of bacteria that have been transformed with a recombinant plasmid compared to nontransformed cells.Some of the most common selectable markers are genes for ampicillin resistance (ampR) and tetracycline resistance (tet R ) and the lacZ gene used for blue white selection. 9) Insertional inactivation refers to the loss of activity of the selectable marker genes due to the insertion of foreign DNA within the coding sequence of the marker gene in a transfected bacteria.

3-Marks Questions:
1)Enlist the major steps in recombinant DNA technology.

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2)Mention the steps involved in the separation and isolation of DNA fragments through agarose gel electrophoresis. 3)Describe in brief the principle of DNA isolation through gel electrophoresis. 4)Highlight the salient features that are required to facilitate cloning into a vector. 5) Enumerate the major steps for isolation of DNA. 6) Draw a neat ,labeled diagram of (a) simple stirred tank bioreactor/ (b) sparged tank bioreactor. HINTS: 1) R-DNA Technology: Restriction enzyme cuts double stranded DNA at its particular recognition sequence.

The cuts produce DNA fragments with cohesive ends

DNA from a plasmid was also cut by the same restriction enzyme

When two of the above mentioned DNA come together they can join by base pairing.

DNAligase enzyme is used to unite the backbones of the two DNA fragments ,producing R-DNA 2)Agarose gel electrophoresis: Electrophoresis Electrophoresis is a technique used to separate and sometimes purify macromolecules especially proteins and nucleic acids - that differ in size, charge or conformation.

Linear DNA fragments of different sizes are resolved according to their size through gels made of polymeric materials such as polyacrylamide and agarose. When charged molecules are placed in an electric field, they migrate toward either the positive or negative pole according to their charge. In contrast to proteins, which can have either a net positive or net negative charge, nucleic acids have a consistent negative charge imparted by their phosphate backbone, and migrate toward the anode

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DNA is electrophoresed through the agarose gel from the cathode (negative) to the anode (positive) when a voltage is applied, due to the net negative charge carried on DNA

When the DNA has been electrophoresed, the gel is stained in a solution containing the chemical Ethidium bromide Ethidium bromide is a fluorescent dye that intercalates between bases of nucleic acids and allows very convenient detection of DNA fragments in gels under U-V radiations, This compound binds tightly to DNA (DNA chelator) and fluoresces strongly under UV light allowing the visualisation and detection of the DNA.

Finally,, the separated DNAs are cut out from the gel and extracted from the gel piece(elution).

3) DNA When charged molecules are placed in an electric field, they migrate toward either the positive or negative pole according to their charge. In contrast to proteins, which can have either a net positive or net negative charge, nucleic acids have a consistent negative charge imparted by their phosphate backbone, and migrate toward the anode DNA is electrophoresed through the agarose gel from the cathode (negative) to the anode (positive) when a voltage is applied, due to the net negative charge carried on DNA 4)Salient features of a DNA cloning Vectors:

Size: small enough to be easily separated from the chromosomal DNA of the host bacteria. Ori site; must have the site for DNA replication that allows the plasmid to replicate separately from the host cells chromosome. Multiple Cloning sites :a stretch of DNA with recognition sequence for many different commonb restriction enzymes. Selectable marker genes RNApolymerase promoter sequence

Major steps for isolation of DNA:

Cell containing DNA is treated with lysozyme/cellulose/chitinase

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DNA along with RNA,Protein,lipd are released

Treatment with RNAase,protease to remove RNA And Protein

Appropriate treatment to remove other impurities

Addition of chilled ethanol to get precipitation of purified DNA 6) Consult NCERT Textbook page number 204

5-Marks Questions:
1) What do you mean by PCR? Briefly enumerate the major steps of PCR. Mention the utility of PCR. HINTS: PCR is a cycle of three steps: 4. DENATURATION - the strands of the DNA are melted apart by heating to 95C 5. ANNEALING - the temperature is reduced to ~ 55C to allow the primers to anneal to the target DNA 6. POLYMERISATION/EXTENSION - the temperature is changed to the optimum temperature in order for the DNA polymerase to catalyse extension of the primers, i.e. to copy the DNA between the primers. The cycle is repeated over and over again - as many times as needed to produce a detectable amount of product (DNA)

HIGH ORDER THINKING SKILLS (HOTS).

1) Why dont restriction enzyme digest chromosomal DNA in bacterial cells ? 2) Why do bacteria have plasmids? 3) Why thermostable DNA polymerase is essential in PCR?

4)Eukaryotes do not have restriction endonuclease, then how they manage with normal endonuclease enzyme? 5) It is advisable to use different restriction endonucleases to cut the vector DNA and source DNA.Why ? 3 Uncontrolled recombinant DNA technology experiments is dangerous to mankind. Comment on it.

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4)

+
Foreign DNA + plasmid

=
??

Complete the above sequence of diagrammatic representation and name it. 5 ). (a) Which is the most commonly used matrix in gel electrophoresis ? (b) What is the source of it? 6) Find the odd one out and write why that is odd (a) Sal I, Pst I, Cla I, BamH I, pBR 322 (b) Bacteria, Virus, Gene-gun, Fungi 7) Detect the mismatch from the following and replace the wrong match with a right one (a) ECOR I Bacteria (b) Ethidium Bromidqe- Gel electrophoresis

(c) Lysozyme- Fungi (d) Palindrome sequence-Restriction enzyme

8). Name the enzyme involved in the following process: (a) Repeated amplification of DNA fragments. (b) Formation of short piece of RNA strand for annealing. (c) Breaking of bacterial cell to release DNA and other macromolecules. (d) Cutting and rejoining DNA fragments.

(e) Formation of m-RNA

(f) Joining of foreign DNA fragments with plasmid. 9)Explain how recombinants and non- recombinants are differentiated on the basis of colour production in the presence of a chromogenic substrate. Name that procedure.

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Chapter-12 BIOTECHNOLOGY & ITS APPLICATION

Biotechnology has given several useful products using microbes, plants, animals & their metabolic machinery. Applications- i) Diagnostic & therapeutic ii) Genetically modified crops iii) Waste treatment iv) Energy production v) Food processing vi) Bioremediation Application in agriculture Genetically modified organisms (GMO)-Plants, bacteria, fungi, animals.Whose genes are altered by manipulation. Transgenic crops-Crops contain or express one or more useful foreign genes. Advantages-i) More tolerant to stresses (heat, cold, drought). ii) Pest resistants GM crops, reduced the use Chemical pesticides. Eg- BT-Cotton iii) Reduced post harvest losses. Eg- Flavr savr tomato. iv) Enhance nutritional value of food. Eg- Golden Rice (Vitamin A enriched). v) Increased efficiency of mineral uses. Bt- cotton -- BT stands for Bacillus thuringenesis (Soil Bacteria). Bacterium produces proteins (Crystal Protein) that kills the insects. Remains as Protoxin (inactive) in bacteria, converted to toxins in alkaline medium (i.e. in the gut of insects) and cause death of the insect. Protection of plants against nematodes Nematode, Meloidogyne incognita infects tobacco plants & reduces yield. Specific genes (DNA) from nematodes introduced into the plants using Agrobacterium. Genes produce sense and antisense Complementary RNA. Complementary RNA neutralizes the specific RNA of nematodes by a process called RNA Interference and parasite cannot live in transgenic host. In medicine- genetically engineered insulin Human insulin consists of Polypeptides chain A & B. Insulin secreted as Pheromone, which contains C peptides, and it is removed during maturation. In 1983, Eli Lilly, an American company prepared 2 DNA sequences coding for chains A & B. Gene therapy Genes inserted into the cells and tissues to correct certain hereditary diseases.

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ADA (Adenosine Deaminase) defiency can be cured by bone marrow transplantation. Molecular diagnosis -- PCR (Polymesase chain reaction) used for early diagnosis of disorder. Elisa (Enzyme Linked Immunosorbent Assay) used to detect AIDS Biropiracy -- Some organizations and multinational companies exploit or patents bioresources of other nations without proper authorization. 1) Name the bacterium that was the first biopesticides used on commercial scale in the world? ANS- Bacillus thuringenesis 2) What do the terms Cry and cry represent respectively? ANS- CryProtein, cryGenes 3) What does ELISA stand for? ANS- Enzyme Linked Immunosorbent Assay 4) Which Nematodes infects the roots of tobacco plants and causes a great reduction in yield? ANS- Meloidegyne incognita 5) What is DNA Probe? ANS- Short specific artificially produced segments of DNA. 6) Name the disease for which first chemical gene therapy was given? ANS- ADA deficiency to a 4- years girl in 1990. 7) What is Alpha-lactalbumin? Where is it produced in human body? ANS- Important human milk protein found in human breast milk. 8) Crystal of BT toxin produced by same bacteria kill the bacteria themselves. Why? ANS- It remains as protoxin (inactive form) in bacteria. It becomes toxin in alkaline medium ( in gut of insects ). 9) Expand the terms :GEAC, SCID ANS- GEAC- Genetic Engineering Approval Committee. SCID- Severe Combined Immuno Deficiency 10) Which transgenic animal is used for testing the safety of Polio vaccine? ANS- Mice. 169

11) Name the first transgenic cow? ANS- Rosie. 12) What are genetically modified organisms? ANS- Organisms (plants, bacteria, fungi, animals) whose genes have been altered. 2marks questions 1) What was the speciality of the milk produced by the transgenic cow Rosie? Ans- i) Contained alpha lactalbumin ii) Nutritionally more balanced than normal cow milk 2)Name some techniques used for early molecular diagnosis of pathogens and genetic disorders? Ans- i) rDNA technology ii) PCR (polymerase chain reaction) iii) ELISA (enzyme linked immune sorbent assay) 3) Explain why children eating golden rice are unlikely to suffer from nightblindnes? Ans- i) It is genetically engineered with high levels of beta carotene and other carotenoids which enhance vitamin A in rice . ii) Lack of vitamin A causes night blindness . 4) Describe briefly how does Hirudin protein is being produced through a transgenic plant . Ans- Gene encoding Hirudin chemically synthesized and transferred to brassica napus where it accumulates in seed and purified and used as medicine. Hirudin protein stops blood clotting. 5) What is meant by RNA interference ? Explain . Ans- Silencing of a specific mRNA due to complementary ds RNA molecule that bind to and prevents translocation of the mRNA (silencing) 3marks questions 1) What are the potential risks ( Three ) and benefits(Three) of GM crops ? Ans Potential risks- i) Products of transgene - allergic or toxic ii) Cause damage to natural environment iii) Weeds also become resistant iv) Can endanger native species Benefits i) More tolerant to abiotic stress. ii) Disease resistant .

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iii) Enhanced nutritional value 2) What are cry proteins? Name the organism that produces it . How has man exploited this protein to his benefit? Ans i) Protein crystals containing toxic insecticide. ii) Produced by soil bacterium , Bacillus thuringenesis. iii) BT toxic genes isolated and incorporated into several crop plants ( BT cotton , BT corn ) to provide resistance against insect pests. 3) What are probes ? How it is used in diagnosis of diseases? Ans Probes are small (15-30 bases ) nucleotide sequences used to detect the presence of complementary sequences in nucleic acid samples. Single stranded DNA or RNA with radioactive molecules allowed to hybridize its complementary DNA in clone cells . Followed by detection using autoradiography. Clone containing unitated gene appear on photographic film.

5 marks questions 1)Explain the steps involved in the production of genetically engineered insulin? Ans- i) Human insulin consists of 51 amino acids arranged in chains of A and B bearing 21 and 30 a.a respectively interconnected by disulphide bridges.

Diagram- Maturation of proinsulin into insulin after removal of c- peptide

ii) Insulin synthesized as prohormone has extra c -peptide which is removed during maturation.

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iii) In 1983 , Eli Lilly, American company prepared two DNA sequences similar to A and B chains of human insulin(humulin). iv) Chain A and B extracted and combined by creating disulphide bonds. 2) a) Name the nematode that infects and damages tobacco roots . b) How are transgenic tobacco plants produced to solve this problem. Ans a) Melodegyne incognitia b) i) Nematode specific genes isolated , cloned and introduced into tobacco plants ii) ds RNA are produced that inititated RNA interference . iii) mRNA translation silenced. iv) Survival of the nematode not possible in the transgenic host plant. 3) Describe with examples why transgenic animals are produced ? Ans-i) Transgenic animals produce useful compounds , created by introducing a porton of DNA that code for the products. Example :-Alpha 1 antitrypsin used for treating emphysema,. ii) These animals help to test the safety of vaccines Ex:- Polio vaccine is tested on mice iii) More sensitive to toxic substances. iv) It increases our understanding as to how genes could control the development of diseases. v) The study of regulation of genes and how it affects the normal function. Example:-Information is obtained about how insulin act as a growth factor .

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Chapter 13 ORGANISMS AND POPULATIONS

Capsule Levels of organisation 1.organism 2.population 3.biotic community 4.ecosystem 5.biome 6.biosphere Environment a) abiotic factors Temperature Water Light Soil b) RESPONSE TO ABIOTIC FACTORS Regulate Conform Migrate Suspend Adaptation population growth and growth forms change Nt=N + B+I D+E

characteristics growth rate

GROWTH CURVE J or S Shaped dN=rN dt Change in population size

POPULATION ATTRIBUTES POPULATION GROWTHImmigration + Population density Emigration

Growth ModelsExponential growth Logistic growth Population interactionsPositive mutualism commensalism negative competition predation parasitism ammensalism

Density dN dt = rN K-N K

Natality

Mortality

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13. organisms and populations Ecology # It is a branch of science. # It deals with the interaction (i) Among organisms (ii) Between organisms (iii) Physical environment. # Deals with four levels of organization

Levels of organisaton BIOSPHERE

BIOME

COMMUNITY

POLULATION

ORGANISM

Organism Individual of specific species e.g. cat, rat, human being, etc. Population # Group of individuals. # Observed intra-specific competition for basic needs. Community # Aggregation of populations # Observe both Intraspecific and Interspecific competition. Biome # Combination of various communities. # Regulation type and associated fauna in specific climatic zone. # Seasonal variation and annual variation lead to biome formation # E.g. Artic and Alpine tundra, coniferous forest, temperate forest, grass land and desert. Biome distribution

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Environment

Abiotic factors Temperature, soil, water, light

Biotic factors Microorganisms, plants, Animals

Temperature # Average temperature varies seasonally # Decreases from equator to poles and plains to mountains # Temperature ranges from sub Zero to 50 C in tropical forests # Function and distribution of organisms temperature dependent. # Organisms Eurythermal or Stenothermal # Organisms affected by Global Warming. Water # Influences life of organisms. No life without water. # Productivity and distribution of plants water dependent. # Quality important for aquatic organisms #Organisms Euryhaline or Stenohaline. Light # Photosynthesis and release of oxygen light dependent. # Sciophytes need use diurnal and seasonal light intensity of forage, migration and reproduction. # Distribution of algae at different depths light dependent. Soil # Nature and proportion of soil in a place depends on climate, weathering process and types of soil. # Soil composition, grain size and aggregation determine percolation and water holding capacity of soil. # Physical and chemical properties determine type of plants and Animals that survive in a habitat. # Biotic sediments determine type of Benthic Animals.

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Response to environmental condition

Regulation

suspention

organism

conformation

migration

Regulation # Organisms maintain homeostasis achieved by physiological and behavioral means # Have constant body temperature (Thermoregulation) # Constant osmotic concentration (Osmoregulation) Conformation # Cannot maintain constant internal Environment # Body temperature changes with the ambient temperature # Osmotic concentration of body fluid changes with the ambient concentration of medium. # Thermoregulation energy expensive, heat loss or gain is a function of surface area of body. Migration Occurs in stressful condition Organism moves away temporarily to another habitat. Birds undertake long distance migration. Susppension Organisms suspend their metabolic activities during stressful condition Resume their function at the return of favorable conditions. E.g. Hibernation of Frog, Reptiles, Polar Bear etc Aestivation in Snail and Fish. Seed dormancy. Adaptation Morphological, physiological and behavioral changes that enable organisms to adjust to the Ever changing environment E.g. Kangaroo rat survives in desert conditions through internal oxidation of fat, removing concentrated urine of les quantity. Allens rule-cold climate mammals have shorter ears and limbs to minimize heat loss. Polar mammals like seals have blubber to prevent heat loss. Burrowing habit to escape form heat Higher count of RBC, Hb at high altitudes. Population attributes *Birth Rate *Death Rate *Sex Ratio 176

*Population density. Age pyramids # Three ecological ages: # Pre-reproductive, Reproductive and Post-Reproductive # High proportion pre-reproductive individuals occur in expanding population # Pre-reproductive individuals are uniform in stable population. # Pre-reproductive individuals are less in Declining population.

Representation of age pyramids for human population

Post Reeproductive

Reproductive

Pre Reproductive EXPANDING Population growth Factors that affect the size of population Food availability Weather Predation pressure Competition Density of population at any time at a given place depends on Natality, Mortality, Emigration Immigration STABLE DECLINING

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Population growth models

Exponential growth : dN/dt rN Logistic growth : dN/dt rN K N/K Where dN = Difference in the number of individuals in a population. dT = Difference in time. rN = intrinsic rate of natural increase in number of individuals K = carrying capacity of the area N = number of existing individuals in a population.

Factors that affect population density

IMMIGRATION I

NATALITY B

POPULATION DENSITY N

MORTALITY D

EMIGRATION

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Types of population interactions Interaction Species a Mutualism Predation Parasitism Commensalism Competition Ammensalism

Species b

Mutualism
Both the species get benefited. Lichens Relationship between Non-photosynthetic Fungus and photosynthetic Algae or Cyanobacteria.

Mycorrhiza Asociation between Fungui and Higher Plants like Pinus. Plants and insects for pollination Orchid ophrys and male bee a good example for co-evolution of plants and Animals. PREDATION One species get benefited and the is harmed. Tiger and Deer

Snake and Frog Herbivores and plants Competition Both the species are harmed. Flammingoes and resident fishes compete for the common food zooplankton in

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South American lakes. Abington Tortoise and goats in Galapagos Islands for food. Gouses Competitive Exclusion Principle -Two closely related species competing for the same resource cannot co-exist indefinitely and trhe competitively inferior on will be eliminated eventually. Parasitism One species gets benefit and the other is harmed.

Parasites

Endoparasites Liver fluke, plasmodium

ectoparasites lice, ticks

brood parasites koel

Adaptations of parasites # Loss of sense organs # Presence of adhesive organs or suckers # Loss of digestive system # High reproductive capacity.

Ammensalism One species hurts the other but the other is not affected. Penicillium secretes Penicillin and kill Bacteria but by this Penicillium does not benefit. Algal bloom leads to death of fishes, but the death of fishes is of no use to the algal bloom. Commensalism One species benefits and the other neither harmed nor benefited. The cattle egret catches the insects disturbed by moving cattle, but the cattle neither harmed nor benefited. Another example The clown fish gets protection from predators by close association with sea anemone, but the sea anemone has no effect.

Very short type questions (1 mark) 1. Define Ecology. Ans. Branch of biology, which studies the relationships of living organisms with the abiotic and biotic components of their environment. 2. Name the factors that account for the formation of major biomes. Ans. A) The annual variations in the intensity and duration of temperature and B) The annual variation in precipitation. 3. Most living organisms cannot survive at temperature above 45 C. How are some microbes able to live in habitats with temperature exceeding 100 C.

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Ans. Possess some heat-resistant or thermostable protein/enzymes and an array of biochemical adaptations. 4. Why many of the fresh water animals cannot live for long in sea water or vice versa? Ans. They would face osmotic problems. 5. Define homeostasis. Ans. It is the phenomenon in which the organisms maintain a steady constant environment, despite varying external environmental conditions. 5. Why is the polar region not a suitable habitat for tiny humming birds? Ans. Have a larger surface area to body volume ratio, tend to lose heat fast when it is cold outside, have to spend more energy to generate body energy. 6. What is meant by Allens rule? Ans. Allens rule refers to the minimizing heat loss in animals by possessing shorter ears and limbs. 7. Why do people living in high altitudes have a higher red blood cell count and total haemoglobin? Ans. To compensate for the low atmospheric pressure. Low oxygen content of air prevailing at high altitudes. 8. When does a population growth curve become J-shaped? Ans. Sigmoid growth curve represents logistic growth, Growth is slow initially, becomes rapid , then becomes steady as resources become limiting. 9. What is the ecological principle behind the biological control methods of managing the pest insects? Ans. Predation, interspecific interaction where one animal kills and consumes the other weaker animal. 10. Why is the female mosquito not considered a parasite, though it needs our blood for reproduction? Ans. The female mosquito is not considered a parasite because; it does not take shelter on our body and does not harm the body. Short answer type questions (2 marks) 1.Name the four major biomes found in India? Ans. (i) Tropical rain forest (ii) Tropical deciduous forest (iii) Desert (iv) Sea coast 2. How does light influence the life of living organisms? Ans. Intensity and duration of light influence. Foraging , migratory activities and reproduction. 3. Why are very small animals generally not found in Polar Regions? Ans. Heat loss or gain is a function of the surface area. Small animals have a larger surface area relative to their volume. So loose heat rapidly. So they tend to lose body heat very fast when it is cold outside consequently they have to spend more energy to generate body heat. Considering the cost and benefits of energy expenditure very small animals are generally not found in Polar Regions. 4. List the attributes that populations but not individuals possess. Ans. (i) Birth rate (ii) Death rate (iii) Sex ratio (iv) Age distribution 5. Biomass is a more meaningful measure of population size. Explain with an example. Ans. (i) Population large the total number is not an easily adoptable measure. Counting takes long time or practically impossible (ii) There is no need to know the absolute population size for some investigations..

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(iii) Number may sometimes be misleading e.g. In a given area there are 200 Parthenium plants and a single banyan tree.Here biomass size of the banyan tree is much more than those of 200 Parthenium plants. 6. Why is logistic curve growth model considered more realistic? Explain. Ans. No population of any species in nature can have unlimited resources to permit exponential growth. Since resources are finite and become limiting, there is competition among individuals for the limited resource and eventually only the fit individuals will survive and reproduce. 7. Name the important defence mechanisms in plants against herbivory. Ans. (i) Thorns and spines are most common morphological means of defence. (ii) Plants produce and store certain chemicals which may (a) Make the animal sick (b) Interfere with digestion (c)Be poisonous to kill them. Short answer type questions (3 marks) 1.How do organisms manage with stressful conditions existing in their habitats for short duration? Explain with the help of one example each. Ans. (i) Organism moves away from the stressful area to a comfortable area, returns back when the stressful period is over e.g. Birds from Siberia migrate to Bharatpur Sanctuary in Rajasthan. (ii) Animals which cannot migrate show hibernation during winter or aestivation in summer or enter into diapause. (iii)Bacteria, Fungi and lower groups of plants produce thick walled spores which germinate under suitable conditions. (v) In higher plants seeds and some other vegetative reproductive structure serve the purpose. 2. What is brood parasitism? Give an example. What adaptation has evolved in this phenomenon? Ans. * One species lays eggs in the nest of another bird, lets the host incubate them. e.g. Cuckoo lays eggs in the nest of a crow. The eggs of the parasite resemble with the eggs of the host in colour, size. Reduce chances of the host bird detecting the foreign eggs and ejecting them from nest. 3. Name and explain the kind of interaction in the following. 1. Algae and Fungi in Lichens 2. Head Louse and Humans 3. Hermit Crab and Sea Anemone (i) Interaction of mutualism where the two species are equally benefited. Fungus provides protection, helps in absorption of water and minerals,algae provide food for the Fungus. (ii) This is case of Parasitism where the louse is an ectoparasite. Parasite takes shelter on humans and also derives nutrition. (iii) It is commensalisms where one species is benefited and the other is neither benefited nor affected. Sea Anemone is benefited as it does not have to move to places rich in nutrients, while hermit crab is neither benefited nor harmed.

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Long answer type questions (5 marks) 1.Define the following terms and give an example each. a) Commensalism b) Parasitism c) Camouflage d) Mutulism e) Interspecific competition. Ans. Commensalism. i) interspecific ii) one species benefited, other neither benefited nor harmed, E.g. A clown fish living among sea anemone. b) Parasitism i) interspecific ii) one species takes food and shelter from another iii) damages host iv) parasite benefited host harmed. c) Camouflage Adaptation where the animal blends with the surroundings to escape detection by predator e.g Frogs have olive green skin with patches of chromatophores to camouflage with grass. d) Mutulism Both the species are mutually benefited ,e.g. An alga and a fungus in lichen. e) Inter specific competition. Individuals of two different species compete with each other for certain resources and both are harmed 2.What is an age pyramid? What do they show about human population? Represent diagrammatically the different shapes of age pyramids and mention what each of them represents. Ans. Graphic representation of the age distribution ,i.e. per cent individuals in different age group of a population. For human population, the age pyramids generally show age distribution of males and females in a combined diagram.

Expanding

Stable

Declining

* Shape of the age pyramid reflects growth status of the population. * Size of the population tells us about its status in the habitat. * Ecological process is investigated in terms of a change in the population size.

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CHAPTER-14 ECOSYSTEM A.G.Tansley coined the term ecosystem in1935. Types-a)Terristrial-Forest, grassland, trees, desert etc. b)Aquatic-Pond, lake, river etc. Components a)Biotic Producers, consumers,decomposers b)Abiotic-Air,water,soil,temperature. ii) stratification- vertical distribution of different species in different strata. iii) tropic organization- Food relationship of producers and consumers. 1) Productivity Two types i)primary productivity-I) amount of bio-mass produced per unit area .It is of two types a)GPP(Gross primary productivity) Rate of synthesis of organic matter per unit area . b)NPP(net primary productivity)- NPP = GPP R (respiration ) ii) secondary productivity Rate of formation of new organic matter by consumers . 2) Decomposition- Breakdown of complex organic matter into simpler inorganic substances by decomposers . Steps in decomposition i) Fragmentation of Detritus ii) Catabolism iii) leaching iv) Humification and mineralisation 3) Energy flow Flow of energy is governed by law of thermo dynamics. a. Energy flow unidirection from producers to consumers . b. In each trophic level there is loss of 10 % energy(10%Law) 4) Nutrients cycle or biogeochemical cycle Different nutrient cycles are i) Carbon cycle ii) Phosphorus cycles Specific place occupied by organism in the food chain. It is a division of food chain .

STANDING CROP Biomass present in atrophic level at a particular time. Biomass is the dry weight of organism.

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Food chain is of two types i)Detritus food chain Starts with organic matter or detritus. Detritus Earthworm Death Frog death Snake death Peacock death

Decomposers (detrivores ) ii) Grazing food chain Known as predator food chain. Grass (producer) Goat (P.consumers) Man (S.consumers )

Food web Natural interconnected food chains .i) ii) unidirectional loss of 10 % of energy in each trophic level

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Ecological pyramidsGraphical representation of food or energy relationships between organisms of different trophic levels . Types 3 types -i) Pyramid of numbers Graphical representation of the number of individuals per unit area. It is mostly upright . It may be upside down .

Pyramid of number upside up and upside down. Pyramid of biomass

Graphical representation of biomass per unit area of different trophic levels . May be upside up and upside down.

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Pyramid of energy

Amount of energy trapped per unit time and area in different trophic levels . It is always upside up .

Gradual and fairly predictable changes in species composition of a given area . Types i) primary succession Succesion on bare land ex:- bare rocks ,newly created ponds etc. ii)Secondary succession Occures in an area where there was vegetation earlier. Soil or sediment present that is why it is faster than primary succession . Two types ,based on nature of habitats. i) hydrach succession Succesion in water or wet areas Steps in hydrach Phytoplanktons--- Floating angiosperms---Rooted hydrophytes---sedges---grasses---shrubs and trees (Pioneer) (climax)

(Seral)

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ii) Xearch succession succession on bare rock Stages or steps Lichens---mosses---herbs---shrubs---trees (pioneer) (climax)

(seral) i) Carbon cycle

ii)Phosphorus cycle

Chapter-14 Ecosystem LEVEL1:-

Q1 ) What is biomass ? Ans Total dry weight of organic matter present in an organism . Q2 ) Name the four important functional aspects of ecosystem . Ans i) Productivity ii) Energy flow iii) Decomposition iv) Nutrient cycling

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Q3) Name the two forms of reservoir of carbon that regulate the ecosystem carbon cycle . Ans Oceans and atmosphere . Q4) Name the dominant producers in a deep aquatic ecosystem . AnsPhytoplankton. Q5) What is NPP? ANS NPP= GPP-R

Q4. What are the two basic catagories of ecosystem? Give example. Ans-a) Terrestrial-Forest, grassland, desert. b) Aquatic-Pond, lake, sea, ocean Q5.What is food chain? Give an example. Ans-a) Food and feeding relation among organisms makes a chain like structure b) GrassDeerLion Q6. Describe the major components of ecosystems. Ans- a) Biotic-i) Producer-green plants. ii) Consumers-primary, secondary,tertiary and decomposers. b) Abiotic-i) Physical and climatic factors-soil, temperature,light, humidity. ii) Chemical factors-inorganic chemical substances (sodium, potassium, nitrogen etc.) organic substances-(humus, protein, fat etc.)

LEVEL 2 Q1) What do you mean by transducers ? Ans Producers are called transducers because producers change light energy into chemical energy . Q2) What is meant by productivity of a trophic level ? Ans - Rate of synthesis of energy containing biomass per unit area in unit time . Q3) Which species is the pioneer species on a bare rock Ans Lichen 189

Q4.What are decomposers? Write their function. Ans-a)Saprotrophs feed on dead bodies of organisms, b) Decomposition and mineralization. Q5.What is the difference between gaseous and sedimentary cycle? Ans-a) Gaseous-Reservoir in atmosphere ,Nitrogen cycle b) Sedimentary-Soil,e.gphosphorus. Q6.Mention two factors by which productivity is limited in an aquatic ecosystem. Ans-a) Light-decreases with increasing water depth. b) Nutrient Limiting factor in Deep Ocean Q7.Expand PAR, How much PAR is used in gross primary productivity? Ans-a)PAR-Photosynthetically active radiations b) 2-10% is used. Q8.Give account of factors affecting the rate of decomposition. Ans-a) climatic factor i)temp ii) soil ) chemical quality of detritus Higher temp and moist condition high rate of decomposition Dry soil , High temp Low rate Q9) What are ecological pyramids ? Mention its limitations . Ans a) Arrangement of trophic levels from producers to top carnivores forms pyramid like structure 3 types i) Pyramid of number ii) Biomass iii) Energy Limitations i) Assumes simple food chain iii) Single species may operate at two or more trophic levels. Q10 ) Explain carbon cycle with ray diagram . Ans Given in text. Q11. Give an account of energy flow in an ecosystem. Ans- Rate of energy transfer between the organisms of different trophic levels is called energy flow. Energy flow is unidirectional, 10% loss of energy in each trophic levels. 190

2-10% PAR captured by green plants.

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Energy flow diagram from the text.

LEVEL 3

Q1) Why the pyramid of energy is always upright ? Ans Because when energy flows from one trophic level to another trophic level , always some energy is lost as heat .

Q2) What is bioenergetics ? Ans Transformation of solar energy into chemical energy by producers . Q3.Why is the length of a food chain in an ecosystem generally limited to 3-4 trophic levels? Ans As 90% energy is lost in the from heat from one trophic level to another, residual energy decreases drastically within 2-3 trophic levels. Q4.What are the differences between detritus and grazing food chains? Ans-a) Detritus-dead and decaying organic matter. b) Grazing-Living green plants.

Q5.Briefly describe the process and products of decomposition. Ans-Breakdown of complex organic matter by decomposers. a)Process-i)fragmentation ii)leaching iii)catabolism. Humification and mineralization humification leads to accumulation of dark colour substance called humus. Mineralisation result in release of inorgranic substances. b

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Q6 .Describe pond as an ecosystem . Ans- Pond has biotic and abiotic components a) Biotic Phytoplankton ,Zooplankton , small fishes , large fishes , frogs , snake ,etc. b) Abiotic - water , dissolved organic and inorganic substances ,sunlight , temp . Phytoplankton (microscopic plants ) producers . Zooplankton (microscopic animals ) primary consumers Small fishes - secondary consumers Large fishes , frog, snails tertiary consumers Q7.What is xerosere? Describe the process of succession on a bare rock. Ans-a) Succession on bare rock. b) Steps in Xerosere i) Lichens-Pioneer Community. ii) Mosses iii) Herbs iv) Shrubs v) Trees-Climax community.

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Chapter 15 BIODIVERSITY AND CONVERSATION

Biodiversity is defined as the totality of genes, species and ecosystems of a given region,i.e.the combined diversity at all levels of biological organisation.

GENETIC DIVERSITY

SPECIES DIVERSITY BIODIVERSITY

ECOLOGICAL DIVERSITY

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More than 70% of all the species are animals. Among animals insects are the most species rich taxonomic group. Plants comprise no more than 22% of all species. 1) Lattitudinal gradients species diversity decreases as we move from equator to poles.e.g. Columbia (near equator) has 1400 species of birds and Greenland (at 71degree North) has only 56species . Speciation is generally a function of time. Temperate region is subjected to glaciation.TropicaL regions have remained undisturbed . 2) SPECIES AREA relationships :

S=CAZ

LogS=logC+ZlogA

ALEXANDER VON HUMBOLDT observed within a region species richness increased with increasing explored area but only up to a limit. The relation beween species richness and area for a wide varity of taxa turns out to be a rectangular hyperbola. On a logarithmic scale the relationship is a straight line as in a eqution LogS = logC +Z log A

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Where S= species richness,A = Area,Z = slope of the line(regression coefficient),C = Y- intercept Loss of biodiversity Loss of biodiversity in a region may lead to 1)decline in plant production 2)lowerd resistance to environmental changes such as drought. 3)increased variability in certain ecosystem proesses such as plant productivity, water use,pest & disease cycles. Four major causes of biodiversity loss i)Habitat loss and fragmentation ii)over exploitation iii)Alien species invasions iv)Co-extinctions Biodiversity conservation Reasons for conservation can be grouped into three categories: a)narrowly utilitarian-for deriving direct economic benefit from nature. b)broadly utilitarian-as biodiversity plays a major role in many ecosystem services. c)ethical-we need to realise that every species has an intrinsic value and we need to pass on our biological legacy to future generations in good order. How to conserve biodiversity: IN SITU CONSERVATION Threatened /endangered plants and animals are provided with urgent measures to save from extinction in wildlife sancturies, national parks & biosphere reserves, sacred groves. Biodiversity hotspots regions with very high levels of species richness.e.g.western ghat, Himalaya. EXSITU CONSERVATION Threatened animals & plants are taken out from their natural habitat & placed in a setting where they can be protected and given care as botanical gardens, zoos, seed/pollon banks, gene banks etc.

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Chapter 16: Environmental Issues

Pollution Any undesirable change in physical, chemical or biological characteristics of air, land, water or soil which harms the human beings. POLLUTION

AIR POLLUTION Pollutants

WATER POLLUTION

SOIL POLLUTION

NOISE POLLUTION

Agents that bring about pollution E.g. smoke, dust, pollen, chemical pollutants, wastes from hospitals, E-wastes etc. Biodegradable and non -biodegradable pollutants Ways of removing particulate matter 1. Electrostatic Precipitator Discharge corona

Negatively charged wire

Dirty Air

Clean air

Dust particles

Collection plate grounded

Electrostatic precipitator

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2. SCRUBBER

3. Proper maintenance of Automobiles Advantage of cng over diesel CNG burns most efficiently Cheaper Cannot be siphoned, Cannot be adulterated. Problems in use of cng Difficulty in laying down pipelines Non-assurance of uninterrupted supply Steps taken in delhi to reduce pollution Phasing out old vehicles Use of unleaded petrol Use of low sulphur Petrol and Diesel Use of catalytic converters in vehicles Application of stringent pollution level norms for vehicles.

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Noise pollution It is undesirable high level of sound. Harmful effects of noise pollution Psychological and Physiological disorders Damage of eardrums and hearing ability Cause Sleeplessness, increased heartbeat altered breathing pattern, stress etc. Steps to be taken to control noise pollution Use of sound absorbent materials or by muffling noise in industries Demarkation of horn free zones around hospitals and schools. Permissible sound levels of crackers, Timings after which Loudspeakers cannot be played Water pollution Biological Oxygen Demand (BOD) indicates the rate of uptake of oxygen by microorganisms in a sample of water. It shows the presence of organic matter in water, Greater the BOD more is the polluting potential. Less is BOD, less oxygen will be consumed if all organic matter in a litre of water is oxidized by Bacteria completely. Effects of bod Bacteria consume dissolved oxygen for biodegradation of organic matter high in downstream areas and it causes of mortality of fish and other aquatic organisms. Algal bloom It is free floating (Planktonic) Algae. Imparts a distinct colour to water bodies Cause deterioration of water quality and fish mortality. Some blooms are toxic to humans and Animals. Water hyacinth (Eichornia crassipes) Worlds most problematic aquatic weed Called as Bengal Terror Grows faster than our ability to remove.

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Biomagnification Increase in concentration of the toxicant at successive tropic levels Biomagnification of ddt Water 0.0003 ppm Zooplankton 0.04 ppm Small Fish 0.5 ppm Large Fish 2 ppm Fish-eating Birds 5 ppm

Aquatic food chain Eutrophication Natural ageing of lake by biological enrichment of its water. Cultural or accelerated eutrophication Acceleration of ageing process of a lake by effluents from industries and homes. Integrated waste water treatment in arcata It consist of two steps a) Conventional sedimentation, filtering and chlorine treatment, b) Passing this water through marshes for neutralization absorption and assimilation of pollutants. c) Upkeep of this project by FOAM (Friends Of Arcata Marsh). SOLID WASTES

Biodegradable

Recyclable

Non-Biodegradable

Ecological sanitation (Ecosan ) A sustainable system for handling human excreta without using water but with composting method. Advantages of ecosan a) Wastage of water is reduced b) Practical and efficient c) Hygienic and cheap d) Excreta can be recycled and used as natural fertilizer.

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Hospital wastes Syringes, discarded medicines, Used gloves, Post operative materials etc Should be treated before disposing off. E-wastes Unused or damaged computers, calculators, mobile phones etc Developed countries have plants for recycling e-wastes for recycling of metals. In developing countries e-wastes are buried in landfills or incenerated. Agro chemicals Chemicals used in agricultural fields. Fertilizers, pesticides, weedicides etc. They are toxic to even non target organisms. Excess fertilizers cause Eutrophication. They cause soil pollution Advantages of organic farming Economical Wastes do not get accumulated but recycled Does not cause Eutrophication Radioactive wastes Emit radiations and damage biological organisms. Nuclear wastes are called potent pollutants, as they are lethal even in lower doses. Disadvantages of nuclear plants May happen accidental leakages Unsafe disposal of radioactive wastes Radiation emitted cause mutations in organisms Radiation causes genetic disorders

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Green house effect Earths atmosphere acts as a cover, which allows heat and light to enter in, but heat is not escaped thus warming up the earth. Green House Gases:-Carbon dioxide, methane etc.

Ozone Triatomic molecule of oxygen. Found in stratosphere of atmosphere. CFCs discharged from lower atmosphere move upward UV rays act on these CFCs and release chlorine atoms. Chlorine degrades ozone and release molecular oxygen This process is irreversible and thus ozone is depleted.

Ozone hole

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Soil erosion The removal of top fertile layer due to human activities Reasons: Over cultivation Over grazing Deforestation Improper irrigation practices Waterlogging The crops may droop Leads to salinity of the soil. Slash and burn agriculture/jhum cultivation Farmers cut down the trees of the forest and burn the plant remains. Ash is used as fertilizer and land is used for farming or cattle grazing Later, Land is left uncultivated for several years for replenishment of minerals Effects of deforestation Leads to global warming due to excess carbon-dioxide Loss of biodiversity Damage to hydrological cycle Leads to soil erosion Desertification of land Reforestaton Restoring forest that was existing earlier E.g. Observing Van-Mahotsavas It also occurs naturally Aforestation Developing a forest in a new area where no such forest existed in that area.

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A case study of peoples participation in forest conservation A king of Jodhpur wanted to arrange wood for his new palace in 1731. Few Bishnois hugged the trees and asked to cut them first rather than cutting trees. 365 persons lost their lives in this act A small temple is now present there in remembrance of this act Amrita Devi Bishnoi Wild Life Protection Award is instituted for individuals of rural areas who take keen interest in protecting wild life. Chipko movement It was started by local women of Garhwal, They hugged the trees to protect them from the axes of contractors. Joint forest management (jfm) Started y Government of India in 1980 Local communities worked with the government to save the forest. Communities get forest products for encouragement. Chapter-16 Environmental issues LEVEL 1 1. Define eutrophication. Ans. Nutrient enrichment in water bodies leading to depletion of oxygen and loss of life supporting environment. 2. What is biomagnification? Ans. Increase in the concentration of certain toxic chemicals at succesive trophic levels. 3. What is BOD? Ans. Biological Oxygen Demand is the measure of organic matter in any water sample. 4.Which is the worlds most problematic weed, also known as terror of Bengal Ans. Eichornia crassipes (Water hyacinth) Differentiate between biodegradable and non-biodegradable wastes.

Biodegradable wastes

Non-Biodegradable wastes

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*Can be broken down into harmless simple compounds by the action of decomposers. *Can be used as manure *Cause little pollution

*Cannot be broken down by microbes and get accumulated in the biosphere *Enter the food chain *Cause biomagnifications

5. Describe Chipko Movement. Ans. It was launched in Garwhal, Himalayas by Sh Sunder Lal Bahuguna in 1974. Local women showed enormous bravery in protecting the trees from the axes of the contractors by hugging them. 6. Mention harmful effects of noise pollution on human health. Ans. Stress Altered breathing pattern Increased heart beating and blood pressure Sleeplessness and headache Hearing impairment.

LEVEL 2 1. What is meant by algal blooms? What is its significance? Ans. Excess growth of certain phytoplanlktons due to excess nutrients in water causes Deteriorates water quality, leads to fish mortality. 2 What is Jhum cultivation? Ans Farmers cut down the tress, burn, use cattle for grazing and then allow the land to recover. 3 What is snow blindness? Ans. Inflammation of cornea caused by a high dose of UV-B radiation. 4. Mention the harm caused by fine particulate matter to human beings? Ans. (i) Cause respiratory problems (ii) Irritation of eyes (iii) Inflammation of lungs (iv) Premature death. 5.What are the advantages of Organic farming? Ans. Economical procedure as recycling takes place. Waste not accumulated but recycled Efficiency and utilization of resources increased 204

Does not lead to eutrophication. 6. How do radioactive wastes cause damage to living organism? Ans. Cause mutations in living organisms at a very high rate. Lethal in high doses Causes cancer and other disorders. Reduces the vegetation cover. 7. What measures should be taken to reduce global warming? Ans. Reduce use of fossil fuel Efficient use of energy. Avoid deforestation Reduce human population Control green house gases. 8. Write a short note on ozone depletion. Ans. Ozone found in stratosphere. CFCs discharged from lower atmosphere move upward. In stratosphere UV rays act on these CFCs release chlorine atoms. LEVEL 3 1. What is the effect of DDT in birds? Ans. DDT disturbs calcium metabolism in birds, thinning of egg shell and premature breaking of eggs leads to decline in bird population. 2.. What do you understand by Ecosan? Ans. Ecosan are the toilets which use compositing method for ecological sanitation. 3.. Why are nuclear wastes called potent pollutants? Ans. Because they are lethal even at lower doses and cause damaging disorders. 4. Mention two problems that have arisen due to green revolution. Ans. Water logging and soil salinity. 205

5. What is the effect of DDT in birds? Ans. Disturbs Calcium metabolism Thinning of egg shells and premature breakage of eggs, Decline of bird population. 6. Write an account on Ecological sanitation (Ecosan). Ans. A sustainable system for handling human excreta, using dry composting toilets. Practical, hygienic, efficient and cost-effective solution to human waste disposal Human excreta can be recycled into manure Used in Kerala and Sri Lanka. 7. What is ecological sanitation? What are its advantages? Ans. It is sustainable system for handling human excreta without using water but by composting method. Advantages Hygienic, practical and efficient Conserves water Can be recycled and acts as a natural fertilizer. 8. How can we reduce automobile pollution? Ans. Un-Leaded Petrol- Reduces lead pollution in air. Low Sulphur Diesel- Reduces sulphur pollution in air Four stroke engines to reduce emission of unburnt

hydrocarbons. Tube-Ups to increase air-fuel ratio and help in better combustion. Catalytic Converters to reduce pollution. CNG to reduce pollution and conserve fossil fuels. 9.Mention the adverse effects agrochemicals. Ans. They are toxic to non-target organisms. They cause soil pollution Excess fertilizers cause eutrophication. Chlorine degrades ozone and release molecular oxygen (O3 O2)

In this reaction chlorine acts, as catalyst and loss ozone is irreversible 10..Mention the Supreme Court directions to the Government to reduce pollution. Ans. Switch over to CNG in public transport system 206

Enforcement of Euro II norms for vehicles. Compulsory periodic check up of pollution Use of unleaded petrol Low sulphur petrol and diesel Catalytic converters in vehicles Phasing out of old vehicles. 11. a) Explain the functioning of electrostatic precipitator with the help of a diagram. b) Mention the consequence if the electrostatic precipitator does not work in a power plant. Ans. Used for removing particulate air pollutants. Removes about 99 of the particulate pollutants from the exhaust of thermal power plants.

Electrode wires that are maintained at several thousand volts, which release electrons. Electrons become attached to dust particles giving a net negative charge. Collecting plates are grounded and attract the charged dust particles. Velocity of air between the plates must be low enough to allow the dust particles to fall. If electrostatic precipitator of a thermal plant stops working, all the particulate pollutants get released and pollute the air. Discharge corona

Negatively charged wire

Dirty Air

Clean air

Dust particles

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Collection plate grounded ELECTROSTATIC PRECIPITATOR

Word List for concept mapping POLLUTION POLLUTANTS Undesirable change in environment Agents that cause pollution

WAYS OF REMOVONG PARTICULATE MATTER Electrostatic precipitator Scrubber Controlling vehicular pollution Use of CNG NOISE POLLUTION, ITS EFFECTS AND METHODS OF CONTROL WATER POLLUTION AND ITS CONTROL

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Biological Oxygen Demand Algal Bloom Biomagnification Eutrophication Accelerated Eutrophication WASTE WATER TREATMENT Case study of Arcata FOAM SOLID WASTES Municipal solid wastes Biodegradable, Recyclable and Non-Biodegradable A case study for Plastic Waste E-wastes. A case study of organic farming RADIOACTIVE WASTES GREEN HOUSE EFFECT AND GLOBAL WARMING OZONE EPLETION IN THE STRATOSPHERE Dobson units, CFCs, Snow-blindness, Montreal Protocol. SOIL EROSION AND DESERTIFICATION. WATERLOGGING AND SALINITY DEFORESTATION SLASH AND BURN AGRICULTURE REFORESTATION CASE STUDY OF PEOPLES PARTICIPATION IN CONVERVATION OF FORESTS Amrita Devi Bishnoi Wildlife Protection Award CHIPKO MOVEMEWNT JOINT FOREST MANAGEMENT

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WORD LIST BINGO Chapter-1: REPRODUCTION IN ANIMAL

Life span, reproduction, asexual reproduction, clone, binary fission, buds, zoospores, conidia, gemmules, vegetative propagation, runner, rhizome, sucker, tuber, offset, bulb,vegetative propagules, nodes,sexual reproduction, seasonal flowering, oestrous cycle, fertilization, gametogenesis, isogametes, heterogametes, antherozoid, homothallic, monoecious, hetrothallic, dioecious, staminate,pistillate, unisexual, bisexual, hermaphrodites, meiocytes, pollination Syngamy, zygote, parthenogenesis, external fertilization, internal fertilization, embryogenesis, cell differentiation, oviparous,viviparous,ovary, pericarp. Chapter-2:SEXUAL REPRODUCTION IN FLOWERING PLANTS Androecium, Gynoecium, stamen, filament, dithecous, microsporangia, pollen sacs,tapetum, sporogenous tissue, microsporogenesis, microspore tetrad, pollen grains, exine, intine, germ pores, vegetative cell , generative cell,monocarpellary multicarpellary,syncarpous, stigma , style, ovary,placenta,megasporangia(ovule) funicle, hilum, integuments, micropyle, chalaza, nucellus, embryo sac (female gametophyte), megasporogenesis, megaspore mother cell, megaspore, monosporic, egg apparatus, synergids, antipodals, egg cells, filiform apparatus, polar nucleus, secondary nucleus, pollination, autogamy, chasmogamous and cleistogamous flower, geitenogamy, xenogamy, outbreeding devices, pollen- pistil interaction, artificial hybridization, emasculation, bagging, primary endos- perm nucleus, triple fusion, endosperm , embryo, post fertilization events, scut- ellum, heart shaped embryo, plumule, hypocotyle, epicotyle, plumule, radical, coleorrhiza, coleoptiles, albuminous/nonalbuminous seed, perisperm, seed dormancy, pericarp,false/true /parthenocarpic fruit,apomixes, polyembryony. Chapter-3:HUMAN REPRODUCTION Insemination, implantation, testes, scrotum, seminiferous tubules, sertoli cells, interstitial cells of Leydig, rete testis, vasa efferentia, epedidymis, vas deferens, urethral meatus, accessory reproductive glandsseminal vesicles, prostate, bulbourethral glands, ovaries, oviduct, uterus, cervix, vagina, mammary glands, infundibulum, fimbriae, ampulla, isthmus, perimetrium, myometrium, endometrium,fallopian tube,clitoris,lactiferous duct,spermatogenesis, spermatogonia, primary spermatocytes,secondary spermatocytes, spermatids, spermatozoa, spermiogenesis/spermateleosis, spermiation ,acrosome, semen, oogenesis, oogonia, primary oocyte, primary/secondary follicle, antrum, graffian follicle, zona pellucid, ovulation, polar bodies,menstrual cyclemenarche,corpus luteum, progesterone, menopause,cleavage,morula, blastomeres,trophoblast cells, inner cellmsass, implantation, chorionioc villi, h CG, h PL, estrogen, relaxin, umbilical cord, germ layers-ectoderm, mesoderm, endoderm, stem cells, parturition, foetal ejection reflex, lactation, colostrums. Chapter-4:REPRODUCTIVE HEALTH Health,STDs,AIDS,Reproductive & child health care, amniocentesis, maternal mortality rate(MMR), infant mortality rate(IMR), contraceptive,natural methods, periodic abstinence, withdrawal method/coitus interruptus ,barrier methods, condoms, diaphragms, cervical caps, vaults, intra uterine devices(IUDs), pills, sterilization, vasectomy,tubectomy,Medical Termination of Pregnancy(MTP), Infertility, Assisted Reproductive technologies(ART),in-vitro fertilization(IFV), embryo transfer, test tube baby, zygote intra 210

fallopian transfer(ZIFT),intra uterine transfer(IUT), in-vivo fertilization, gamete intra fallopian transfer(GIFT), Intra cytoplasmic sperm injection(ICSI), artificial insemination(AI), intra uterine insemination(IUI),Adolescence Reproductive and Sexual Health(ARSH), Reproductive Health

Chapter-5:PRINCIPLES OF INHERITANCE AND VARIATION Inheritance, Fillial progeny, factors,traits, homologous chromosome, gene, loci/locus, allele homozygous,heterozygous ,phenotype, genotype, monohybrid cross, dominant , recessive, punnett square,segregation, dihybrid cross, independent assortment,test cross,back cross, incomplete dominance, co-dominance, chromosomal theory of inheritance,linkage, recombination,sex determination, autosomes, sex chromosomes/allosomes,homogametic, heterogametic, mutation, chromosomal aberrations, frameshift mutation, pedigree analysis, consanguineous mating, Mendelian disorders, nondisjunction, haemophilia, sickle cell anaemia, phenylketonurea, in born error of metabolism/metabolic disorder,aneuploidy,polyploidy, syndrome, Downs syndrome, trisomy, Klinefeltors syndrome, gynaecomastia, Turners syndrome, rudimentary/streak gonad.

Chapter-6:MOLECULAR BASIS OF INHERITANCE Deoxyribonucleic Acid(DNA), ribonucleic Acid(RNA),nucleotide, nucleoside, nitrogenous bases(purine, pyrimidine), sugar(pentose-deoxyribose/ribose),Adenine,Guanine, Thymine, Cytosin,Uracil, N-glycosidic linkage, Phosphodiester linkage, double helix, Chargaffs rule, anti parallel polarity, central dogma, histones, octamer, nucleosome, chromatin, euchromatin, heterochromatin, nonhistone chromosomal protein(NHC),Transformation(Griffiths experiment), Transduction (Hershey-Chase experiment), repli- cation,semiconservative replication,template DNA, Meselson-Stahls experiment,replication fork, helicases, topoisomerases, single stranded binding protein, primase,DNA polymerase, DNA ligase,Okazaki fragments, continuous (leading)strand, discontinuous (lagging)strand, Transcription, promoter,structural genes/cistrons,terminator,DNA dependent RNApolymerase,coding strand, polycistronic,monocistronic,coding sequence(exon),non-coding sequence(intron), messengerRNA(m RNA), transferRNA(t RNA),ribosomalRNA(r RNA), Initiation factor, termination factor, Eucaryotic transcription, heterogenous nuclear RNA(hn RNA),capping,tailing, splicing,Translation, genetic code,codon, unambiguous, degenerate, universal, methionine/fMet, START/initiation codon(AUG), STOP/NONSENSE CODON(UAA,UAG,UGA),frame shift (insertion/deletion) mutation, adapter molecule, untranslated region,anti codon, aminoacylation of t RNA, release factor,regulation, gene expression,operon, regulator, promoter,operator,lac-Operon, B-galactosidase,permease,transacetylase,lactose,inducer,switch on/off, inducible system,negative regulation, Human GenomeProject (HGP),GENOME, bioinformatics, DNA sequence, Expressed Sequence Tags(ESTs), Sequence Annotation Bacterial artificial chromosome(BAC),Yeast artificial chromosome(YAC),Single nucleotide polymorphism(SNPs), DNA fingerprinting, DNA polymorphism, repetitive DNA, satellite DNA, Variable Number Tandem Repeats(VNTRs),isolation of DNA, Electrophoresis, blotting, hybridization, probe, autoradiography.

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Chapter-7:EVOLUTION Evolution, Big bang, spontaneous generation, panspermia, Millers experiment, HMS Beagle, CharlesDarwin,fitness,AlfredWallace,fitness,naturalselection,evidences,morphology,anatomy,homologou s organ, divergent evolution, analogous organ, convergent evolution, industrial melanism, adaptive radiation, branching descent, saltation, stabilising selection, directional selection, disruptive selection, gene migration, gene flow, genetic drift, mutation, genetic recombination, founder effect, geological periods, Dryopithecus, Ramapithecus, Homo habilis, Homo erectus, Homo sapiens, Neanderthal man, brain capacity.

Chapter-8:HUMAN HEALTH AND DISEASES Health, disease ,infection, genetic disorders, life style disorders, infectious/ non-infectious, pathogen, typhoid, widal test, pneumonia, common cold, malaria, Plasmodium spp. Anopheles sp. ,Haemozoin, sporozoites, gametocytes, amoebiasis, ascariasis, elephantiasis/filariasis, Wuchereria bancrofti, ringworms, personal hygiene, public hygiene, air borne disease, water borne disease, vector borne diseases, biological control(Gambusia), Aedes, immunity, innate immunity, physical barrier, physiological barrier, cellular barrier, cytokine barrier, acquired immunity, B lymphocytes,T ,lymph ocytes, antibody(Immunoglobuline), light chain, heavy chain, Humoral immune response, cell mediated immunity, active immunity, passive immunity, colostrums, IgA, vaccination, immunization, allergies, IgE, histamine, serotonin, auto immunity, lymphoid organs, bone marrow, thymus, mucosal associated lymphoid tissue(MALT),Acquired Immuno Deficiency Syndrome (AIDS),retro virus, HIV, Enzyme Linked Immuno Sorbent Assay(ELISA),Cancer, contact inhibition, benign tumor, malignant tumor, neoplastic cells, metastatis, carcinogens, viral oncogenes ,proto oncogenes, radiotherapy, chemotherapy, immunotherapy, a-interferon ,drugs, opioids, cannabinoids, cocaine, barbiturates, amphetamines, LSD, hallucinogens, drug abuse, addiction, dependence, withdrawal syndrome, alcohol abuse , liver cirrhosis, danger signs, peer pressure. Chapter-9:STRATEGIES FOR ENHANCEMENT IN FOOD PRODUCTION Animal husbandry, dairy, poultry, animal breeding, inbreeding, out breeding, homozygosity, inbreeding depression, out breeding, out crossing , cross breeding, interspecific hybridization, artificial insemination, Multiple Embryo Transfer Technology(MOET), apiculture, fisheries, plant breeding, green revolution, germplasm collection, cultivars, disease resistance, mutation breeding, insect pest resistance, biofortification, Single Cell Protein(SCP), tissue culture, explants, totipotency, micropropagation, somaclones,meristem, somatic hybrids. Chapter-10:MICROBES IN HUMAN WELFARE Microbes, Lactic acid bacteria(LAB), Saccharomyces cerevisiae, fermentors, distillation, antibiotics, bioactive molecules, streptokinase, clot buster, cyclosporine A, immunosuppressive agents, statins, cholesterol lowering agents, sewage, primary treatment, primary sludge, flocs, Biochemical Oxygen Demand(BOD), Activated sludge, anaerobic sludge digesters, biogas, Ganga action plan, Yamuna action plan, methanogens, Bt cotton, Bacillus thuringiensis, baculoviruses, Trichoderma spp. Integrated Pest Management(IPM), Biofertilisers, organic farming, mycorrhiza, cyanobacteria.

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Chapter-11:BIOTECHNOLOGY:PRINCIPLES AND PROCESSES Biotechnology, genetic engineering, recombinant DNA, gene cloning, gene transfer, origin of replication, plasmid, restriction enzymes, cloning, restriction endonuclease, recognition sequence, nucleases, exonucleases, endonucleases, palindrome, sticky end ,blunt end, gel electrophoresis, elution, cloning vectors, selectable markers, transformation, antibiotic resistance, insertional inactivation, tumor, Ti plasmid, Agrobacterium tumifaciens, micro injection, biolistic/gene gun, lysozyme, cellulose, chitinase, Polymerase Chain Reaction(PCR),denaturation, annealing, extension, thermostable DNApolymerase, bioreactors, downsteam processing. Chapter-12:BIOTECHNOLOGY AND ITS APPLICATION Genetically Modified Organism(GMO), Bt cotton, insecticidal proteins, cry genes, pest resistant plants, RNA interference(RNAi)/RNAsilencing, dsRNA, Genetically engineered insulin, gene therapy, ADA deficiency , c DNA, Molecular diagnosis, transgenic animals, Bio ethics, Genetic Engineering Approval Committee(GEAC), Bio piracy, Indian patent bill. Chapter-13:ORGANISMS AND POPULATIONS Organisms, population, communities, ecosystems, biomes, ecology, grassland,tundra, desert, coniferous forest, temperate forest, tropical forest, abiotic features, temperature,stenothermal, eurithermal, water, salinity, pH, light, soil, moisture,conformers, regulators, partial regulators,migration, suspension, hibernation, aestivation,adaptation,altitude sickness, Allen,s rule population attributes,age pyramid(expanding, stable, declining),population density, natality, mortality, immigration, emigration, exponential growth, logistic growth,mutualism, competition, predation, parasitism, commensalism, amensalism. Chapter-14:ECOSYSTEM Terrestrial ecosystyem, aquatic ecosystem, stratification, productivity, Gross primary productivity (GPP),Net primary productivity(NPP),S econdary productivity,decomposition, detritus, detritivores, fragmentation, leaching, catabolism, humification, humus, mineralization, Photosynthetically active radiaton(PAR),energy flow, producers,consumers,herbivores,carnivores,Grazing food chain(GFC), Detritus Fo0od Chain(DFC), FOOD WEB, TROPHIC LEVEL, STANDING CROP, BIOMASS, Ecological pyramids, upright pyramid, inverted pyramid, succession, pioneer, sere/seral stage, climax community, Hydrarch succession, Xerarch succession, nutrient cycling , biogeochemical cycles, ecosystem services. Chapter-15:BIODIVERSITY AND CONSERVATION Biodiversity, genetic diversity, species diversity, ecological/habitat diversity,mega diversity country, loss of biodiversity,habitat loss, habitat fragmentation, over exploitation, alien/non native/invasive species,co extinctions,narrowly utilitarian, broadly utilitarian,in situ conjservation, endemism, hotspots, sacred groves, Ex situ conservation,Earth summit, sustainable development.

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Chapter-16:ENVIRONMENTAL ISSUES Pollution, pollutants, Environmental Protection Act(EPA), Electrostatic Precipitator(ESP), Vehicular pollution, Compressed Natural Gas(CNG), Euro ii, Bharatii, Air prevention and pollution control act, noise pollution, decibel, water pollution, domestic sewage, dissolved oxygen(DO), Oxygen sag curve, BIOCHEMICAL/BIOLOGICAL OXYGEN DEMAND(BOD), algal bloom, planktic, Bioaccumulation, Biomagnificationj,solid wastes, municipal solid waste, sanitary landfills, plastic waste, e-wastes, agrochemicals, radioactive wastes, enhanced Greenhouse effect , Global warming, CFCs, stratospheric Ozone depletion, deforestation,slash & burn agriculture, Jhum cultivation, deforestation, reforestation, chipco movement, Joint Forest Management(JFM)

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If there was a point mutation in a DNA triplet that changed the code from T T G to T T A, would that cause a problem with the resulting polypeptide chain? Explain Your answer. If a different point mutation changed the DNA code from A C G to A C T, would that cause a problem with the resulting polypeptide chain? Explain Your answer.

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