Lect. 1 Introduction to Plant Biotechnology

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importance of Biotechnology, Branches of Biotechnology, Plant Tissue culture

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Concept of Plant Biotechnology, History of Plant Tissue Culture and Genetic Engineering:

Concept of Plant Biotechnology, History of Plant Tissue Culture and Genetic Engineering Dr. M.K. Karnwal Asstt. Professor, Genetics & Plant Breeding College of Agriculture Biotechnology: An Introduction

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WHAT IS BIOTECHNOLOGY??? BIO – Biological system (s) TECHNOLOGY – Industrial scale venture WHAT FOR? For the benefit of mankind Biotechnology: An Introduction

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Biotechnology: An Introduction Biotechnology: In fact, biotechnology is the product of interaction between science of biology and technology . It is the technological exploitation and control use of biological system. According to the US National Science Foundation: Biotechnology is the controlled use of biological agent such as micro-organism or cellular components for beneficial use. According to the European Federation of Biotechnology: Biotechnology is the integral use of biochemistry, microbiology and engineering science in order to achieve technological (industrial) application of the capabilities of the microorganism, cultured cells and parts thereof.

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WORKING DEFINITION OF BIOTECHNOLOGY Industrial exploitation of biological systems or parts thereof for their commercial utilization in the service of society Biotechnology: An Introduction

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BIOTECHNOLOGICAL ACTIVITY Biochemistry Microbiology Chemical Engineering Natural Product Chemistry Genetics Molecular Biology (Proteomics, Genomics, Metabolomics) Physiology Cell/Tissue Culture Taxonomy Genetic / Recombinant Engineering Plant Breeding Cultivation know how Organic Chemistry Instrumentation Bio-informatics

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BIOTECHNOLOGY PRODUCT Down-stream processing Post harvest processing/storage Packaging and labelling Toxicity/allergy testing Regulatory formalities concerning environment safety Safe transport National/international politics CONSUMERS Biotechnology: An Introduction

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Biotechnology: An Introduction Major Branches of Biotechnology: Medicinal biotechnology Industrial biotechnology Animal biotechnology Environmental biotechnology Plant biotechnology………!

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Biotechnology: An Introduction Medicinal biotechnology S.N. Product Remarks 1. Monoclonal antibodies (used for disease diagnosis e.g. venereal disease, hepatitis B and other viral disease, cancer, etc.) Produced by hybridoma technology 2. DNA probes (used for disease diagnisis e.g. sleeping sickness, malaria, etc.) Produced by genetically engineered microbes . 3. Recombinant vaccines (cleaner, safer e.g. human hepatitis B virus, E. coli vaccines for pigs, rabies virus, etc. Produced by genetically engineered microbes . 4. Valuable drugs like human insulin , human interferon, human and bovine growth hormones , etc. Produced by genetically engineered bacteria

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Biotechnology: An Introduction S.N. Product Remarks 5. Babies of specified sex (artificial insemination with X or Y carrying sperms prepared by sperm separation tech.) It is feared that this may un-favorably change the sex ratio in the population 6. Identification of parents / criminals using DNA finger-printing Very accurate and reliable; from even blood or semen stains, hair roots etc. Medicinal biotechnology

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Biotechnology: An Introduction 2. Industrial biotechnology S.N. Product Remarks 1. Production of useful compounds e.g. ethanol, lactic acid, glycerin, citric acid, gluconic acid, acetone etc. Produced by the microorganisms, mainly bacteria , from less useful substrate. 2. Production of antibiotics e.g. penicillin, streptomycin, erythromycin, mitomycin , cycloheximide , etc. Produced by fungi, bacteria and actinomycetes as secondary metabolites . 3. Production of enzymes e.g. α - amylase, proteases, lipases, etc. From fungi, bacteria etc. for use in detergent, textile, leather, dairy etc. industries & in medicine 4. Fuel production (mainly ethanol, sometimes biogas) from cheap, less useful and abundant substrates e.g. sugarcane bagasse , wood etc. Produced through fermentation by microorganisms . Cowdung -based biogas being popularized in India

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Biotechnology: An Introduction 3. Animal biotechnology S.N. Product Remarks 1. Test tube babies in human ; involves invitro fertilization and embryo transfer. Couple suffering from infertility can have babies. 2. Production of transgenic animals for increased milk, growth rate, resistance to disease , etc. and production of some valuable proteins in milk/ urine/ blood . Transgenic mice, pigs, chicken, rabbits. Cattle, sheep, fish produced. 3. Hormone-induced super ovulation and/ or embryo splitting in farm animals , involves embryo transfer and, in many cases, invitro fertilization. For rapid multiplication of animals of superior genotype.

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Biotechnology: An Introduction 4. Environmental biotechnology S.N. Product Remarks 1. Efficient sewage treatment , deodorization of human excreta. Efficient strains of microorganisms developed. 2. Degradation of petroleum and management of oil spills. A strain of pseudomonas putida. 3. Detoxification of wastes and industrial effluents Genetically engineered microbes. 4. Biocontrol of plant disease and insect pests by using viruses, bacteria, amoebae, fungi, etc. Environment friendly, avoids the use of pesticides, etc. which cause pollution.

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Biotechnology: An Introduction 5. Plant biotechnology S.N. Product Remarks 1. Embryo culture to rescue otherwise inviable hybrids, to recover haploid plants from interspesific hybrids, etc. Applications are the most remarkable. 2. Rapid clonal multiplication through meristem culture, e.g. of many fruit and forest trees, such as, teak. Very high rates of multiplication, conventional rates very low. 3. Recovery of virus and other pathogen free stocks of clonal crops, meristem culture is generally combined with thermotherapy/ cryotherapy. Virus useful in clonal crops, particularly for germplasm exchange.

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Biotechnology: An Introduction 5. Plant biotechnology S.N. Product Remarks 4. Germplasm conservation through storage in liquid nitrogen (at -196*C, cryo preservation) or through slow growth. Particularly useful in clonal crops, especially in those producing tubers, storage roots, etc. 5. Rapid isolation of homozygous lines by chromosome doubling of haploids produced through anther culture/ interspecific hybridization/ ovary culture. Very successful in variety development in China, e.g. in Rice and wheat. 6. Gene transfer (genetic engineering) for insect resistance, protection against viruses, herbicide resistance, storage protein improvement, etc. Mainly using the Ti plasmid of Agrobacterium , also through particle gun, free DNA uptake; REVOLUTIONARY DEVELOPMENT in crop improvement

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WHY WE NEED BIOTECHNOLOGICAL INTERVENTIONS IN PLANTS ?? Biotechnology: An Introduction Plant biotechnology……….!

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Population rise Enhanced diseases Less land for agriculture Malnutrition Desire to learn the secretes of nature We need more plants We need quality plants We need healthy plants We need to conserve plants We need more natural products based medicines Design novel plants superior in traits and productivity Upgrade the genetic machinery through novel gene combination in an otherwise acceptable genomic background Add value to our existing crop usage Demand We must Biotechnology: An Introduction

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Biotechnology: An Introduction History of Biotechnology: The origin of biotechnology can be traced back to prehistoric times when microorganisms were already used for processes like: Fermentation Formation of yoghurt Formation of cheese from milk Vinegar from molasses Production of butanol and acetone from starch by Clostridium acetobutylicum Production of antibiotics from Penicillium notatum .

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Biotechnology: An Introduction History of Biotechnology: Biotechnology is an old science being practiced by nature and man since a long time. Today we are better equipped in terms of knowledge, experimental tools, instrumentation and controls to understand, manipulate and hasten the biotechnological events at cellular, molecular, genetical and biochemical levels on real time scale with precision

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Biotechnology: An Introduction History of Biotechnology: However, the biotechnology got a boost in 1970’s with the discovery of restriction enzyme which led to the development of a variety of gene technology which is considered to be the greatest scientific revolution of the century. Gene Technology/ Genetic engineering: refers to a number of new techniques for changing plants genetically that do not rely on sexual methods but instead involve genetic manipulation at the cellular and molecular level. These are the non-sexual methods of gene transfer.

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Biotechnology: An Introduction A Historical Account of Milestones in the Field of Plant Tissue and Cell Cultur 1904 First attempt to embryo culture of selected Crucifers . Hannig B., Bot. Zeitung, 62: 45-80. 1944 First In vitro culture of tobacco used to study adventitious shoot formation . Skoog F., Am. J. Bot., 31: 19-24. 1949 Culture of fruits In vitro . Nitsch J. P., Science, 110: 499. 1962 Development of MS medium . Murashige T. and Skoog F., Physiol. Plant., 15: 473-497. 1967 Haploid plants from pollen grains of tobacco . Bourgin J. P. and Nitsch J. P., Ann. Physiol. Veg., 9: 377-382 & 10: 69-81.

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Biotechnology: An Introduction A Historical Account of Milestones in the Field of Plant Tissue and Cell Cultur 1977 Successful integration of T-DNA in plants . Chilton M. D. et al., Cell, 11: 263-271. 1985 Infection and transformation of leaf discs with Agrobacterium tumefaciens and regeneration of transformed plants . Horsch R. B. et al., Science, 227: 1229-1231.

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Biotechnology: An Introduction Biotechnology in India: 1982: The government of India constituted a National Biotechnology Board (NBTB) ----To encourage and coordinate research activities in Biotechnology. 1986: A separate Department of Biotechnology was created within the Ministry of Science and Technology .

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Biotechnology in India: Initially, 3 main Research Centre have been organized by the DBT- Indian Agriculture Research Institute Centre (This centre has been named as National Research Centre on Plant Biotechnology ) National Dairy Research Institute Centre Indian Veterinary Research Institute Centre Biotechnology: An Introduction

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Biotechnology in India: DBT created 7 Centers for Plant Molecular Biology ( CPMB ) at the following institutes: Madurai Kamraj University, Madurai Jawaharlal Nehru University, New Delhi Tamil Nadu Agriculture University, Coimbatore Osmania University, Hyderabad (A.P.) Bose Institute, Calcutta National Botanical Research Institute, Lucknow Delhi University (South Campus), New Delhi Biotechnology: An Introduction

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Biotechnology in India: In Addition to the DBT, ICAR, New Delhi and CSIR, New Delhi also promoting research activities in Biotechnology. ICAR has created Biotechnology Wings in their several Research Institutes like- Indian Pulse Research Institute, Kanpur Indian Grassland and Forage Research Institute, Jhansi etc. Biotechnology: An Introduction DBT has focused its attention on the conservation of germplasm using biotechnological approaches -------A National Facility for Plant Tissue Culture Repository has been organized at the NBPGR, New Delhi (3 gene banks created at this centre for MAP) CIMAP, Lucknow TBGRI, Trivandrum

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Biotechnology: An Introduction For the purpose of large scale micropropagation of forest and fruit trees ( eg. Teak, Eucalyptus, poplar etc.) DBT has funded the establishment of three pilot plants at- Tata Energy Research Institute (TERI), New Delhi National Chemical Laboratory, Pune J.N. Vyas University, Jodhpur

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Biotechnology: An Introduction In India DBT also initiated an extensive degree programme for manpower development in biotechnology. DBT Sponsored Regular Teaching Courses in Biotechnology in Different Universities/ Institutes in INDIA University/ Institute Year of Start Annual Intake M.Sc. General Biotechnology ( 2 Year Course) 1. Jawaharlal Nehru University, New Delhi 1985-86 20 2. Madurai Kamaraj University, Madurai 1985-86 20 3. MS University, Baroda 1985-86 20 4. University of Poona, Pune 1985-86 20 5. Banaras Hindu University, Varanasi 1986-87 12 6. Indian Institute of Technology. Mumbai 1987-88 10 7. Roorkee University Roorkee 1991-92 10 8. Aligarh Muslim University, Aligarh 1991-92 10 9. Guru Nanak Dev University, Amritsar 1991-92 10 10. Devi Ahilya Viswavidyalaya , Indore 1991-92 10 11. University of Hedrabad , Hyderabad 1991-92 10

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University/ Institute Year of Start Annual Intake 12. Himanchal Pradesh University, Shimla 1994-95 10 13. University of Calicut, Kerala 1994-95 12 14. Banasthali Vidyapeeth , Banasthali , Rajasthan (for girls only) 1994-95 10 15. Tezpur University, Tezpur (Assam) 1998-99 12 16. Gulburga University, Gulbarga (Karnataka) 1998-99 10 17. Jammu University, Jammu 1999-2000 10 18. Gujrat University, Ahmedabad 1999-2000 10 19. Mysore University, Mysore 1999-2000 10 20. University of Allahabad, Allahabad 1999-2000 12 21. Guru Jambeshwar University, Hissar 2000-2001 10 22. University of Kashmir, Srinagar 2000-2001 10 23. Kumaun University, Nainital 2000-2001 10 M.Sc. Agricultural Biotechnology ( 2 Years) 1. Assam Agriculture University, Jorhat 1988-89 10 2. Tamil Nadu Agriculture University, Coimbatore 1988-89 10 3. GB Pant University of Agri. & Tech., Pantnagar 1988-89 10 4. Birsa Agriculture University, Ranchi 1999-2000 12 5. Himachal Pradesh Krishi Vishvavidhalaya , Palmpur (H.P.) 1999-2000 05 6. Indira Gandhi Agriculture University, Raipur 2000-2001 10 7. Marathwada Agriculture University, Parbhani 2000-2001 10

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Biotechnology: An Introduction Area of Biotechnology: Infect, the classical genetics as applied to plant breeding can not be separated from other improvement in agricultural production. Plant breeding alone may increase up to 50 % of the harvest in the crop cultivation. But, the crop improvement practices of plant breeding have some limitations. Because the plant breeders objective is to improve the efficiency of a crop variety which is different from the force operating in nature where the selection operates at the survival of the fittest rather than efficiency.

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Biotechnology: An Introduction Area of Biotechnology: In past few years a number of methodologies/ technologies have come, that would seems to improve : The photosynthetic efficiency of the plant Nutritional efficiency Resistance ability to pathogens etc. ………. Very faster and quicker as compare to the conventional plant breeding practices. To improve the quality and quantity of the plant through innovative approaches, the two areas are concerned: (1) Manipulation and subsequent growth of cells, tissues, organs and naked plant cells (protoplast) in tissue culture. (2) Second field is genetic engineering/ recombinant DNA technology.

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Biotechnology: An Introduction Area of Biotechnology: (1) Manipulation and subsequent growth of cells, tissues, organs and naked plant cells (protoplast) in tissue culture. (2) Second field is genetic engineering/ recombinant DNA technology. These two areas of research have in recent years become associated with the general field of biotechnology and are potentially applicable to a wide variety of plant species, as well as offering a precision in manipulation of genetic material.

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PLANT TISSUE CULTURE CONCEPTS VOCABULARY APPLICATIONS Biotechnology: An Introduction

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Plant Tissue Culture - Definition The growth and development of plant seeds, organs, explants, tissues, cells or protoplasts on nutrient media under sterile (axenic) conditions. Biotechnology: An Introduction

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Biotechnology: An Introduction Plant Tissue Culture - Definition Based on the culture of part of the plant: plant tissue culture categories into different groups: (1) Plant Culture: Culture of seedlings or larger plants. (2) Embryo culture: culture of isolated plant organs. (3) Organ culture: may be distinguished into different types based on the explant taken for culture from the mother plant. (a) Meristem culture (b) Shoot tip culture (c) root culture etc.

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Biotechnology: An Introduction Plant Tissue Culture - Definition (4) Callus Culture: culture of a differentiated tissue from explant allow to dedifferentiate invitro and a so called callus tissue is produced and this process is termed as callus culture. (5) Cell suspension culture: Culture of isolated cells or very small cell aggregates dispersed in liquid medium (6) Anther culture: Culture of anthers and/ or immature pollen grains in an effort to obtain a haploid cell or callus line. (7) Protoplast culture: culture of plant protoplasts i.e. cells devoid of their retaining cell walls.

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Explant - Definition This means to simply cut-out a very small piece of leaf or stem tissue, or even isolate individual cells, and place them in a tissue culture container. The tissue has to be surface-sterilized so it will not have any contaminating bacteria or fungus. It is then placed inside the tissue culture vessel (dish, jar, etc.) containing a gel called agar. In the agar is dissolved all the sugar, nutrients and plant growth regulators the explant needs. Biotechnology: An Introduction

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Characteristic of Plant Tissue Culture Techniques Environmental condition optimized (nutrition, light, temperature). Ability to give rise to callus, embryos, adventitious roots and shoots. Ability to grow as single cells (protoplasts, microspores, suspension cultures). Plant cells are totipotent, able to regenerate a whole plant. Biotechnology: An Introduction

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PRE-REQUISITES FOR RAISING PLANT TISSUE CULTURES HEALTHY AND YOUNG COMPETENT CELLS WITH MORPHOGENIC PLASTICITY ASEPTIC ENVIRONMENT GROWTH SUPPORTING FOOD SUPPLIMENTED WITH APPROPRIATE HORMONE COMBINATION & CONCENTRATION Biotechnology: An Introduction

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TOTIPOTENCY The capacity of a cell (or a group of cells) to give rise to an entire organism . It implies that all differentiated living cells of a plant, except those that have been exclusively programmed into narrow path of specialization, possess an ability to display full genetic programme and embark upon a developmental pathway similar to that of a zygote, leading ultimately to the formation of a new individual. Totipotency is therefore: Totipotency can be: Genetically (endogenously) controlled Developmentally dependent Strongly regulated by external factors (light, Temperature, wounding etc.) Morphogenic, Physiological, Biochemical, Metabolic and Regenerative Biotechnology: An Introduction

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MORPHOGENESIS Morpho = shapes; genesis = beginning The process of growth and development of differentiated structures from an undifferentiated state of organisation Inception of growth, division, anatomical specialization and multiplication through diverse developmental patterns leading to formation of specialized forms (organs) from earliest to final status. Morphogenesis involves first the fating of cells to undergo morphological change and then carrying out that fating through differentiation and growth. Biotechnology: An Introduction

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DIFFERENTIATION The physiological and morphological changes that occur in a cell, tissue, or organ during development. ORGANOGENESIS The development of tissues and/or organs from individual cells not from pre existing meristems. The formation and development of definite structures/ organs from their primitive conditions (meristemoids/ primordia) REGENERATION Vegetative growth after a wound, amputation or drying of the surface. Process of Growing whole plant from a cell or a group of cells through differentiation and organogenesis Biotechnology: An Introduction

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PLASTICITY The condition of a cell characterized by degree of ease or ready response to a given stimulus for a desired out-put. INDUCTION In its wider developmental sense, is a process seen by its end results. It is the earliest change in the fate or destiny of a cell or a group of cells. It is this change in fate that is what is known as cellular determination. COMMITMENT/ DETERMINATION Event which “initiate” a specific developmental path by singling out from among the various possibilities for which a cellular system might be competent. Biotechnology: An Introduction

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EXPLANT A plant part or piece of tissue(s) excised from a parent plant to initiate a culture SHOOT TIP CULTURE Culture of a shiny, dome-like structure consisting of a shoot apical meristem plus one to few primordial leaves (0.1-1.0mm) MERISTEMOID A localized group of meristematic cells that arise in a cultured tissue (usually callus) and give rise to an organised structure (shoot and/or roots) SOME MORE TERMS IN THE LANGUAGE OF PTC Biotechnology: An Introduction

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CELL GENERATION TIME The interval between consecutive division of a cell. CELL DOUBLING TIME Time required for a cell population to double either in number or cell mass if all the cells in the cultured population are capable of deviding and don’t undergo lysis CLONE A group of genetically identical cells or organisms produced asexually from a common ancestor; they may or may not be homogenous CLONING Asexual multiplication from a single cell/organism. In molecular biology refers to the replication of a small DNA molecule or a gene in a vector Biotechnology: An Introduction

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CELL SUSPENSION CULTURE Culture of single cells or small aggregates of similar cells suspended in liquid medium (Typically single cells in minority and small cell aggregate in majority). BASAL MEDIUM A nutrient support containing all essential elements required growth, a ready made energy (carbon) source and water. It may be defined, undefined, optimal. minimal or selective. CALLUS Actively dividing non-organised tissues of undifferentiated mass and differentiated cells developing from injury (wounding) or in tissue culture. Biotechnology: An Introduction

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UNDIFFERENTIATED Cells existing in a state of cellular development characterized by isodiametric cells, little or no vacuole, large well marked nucleus and lacks specialized structure or function of a cell type in vivo DEDIFFERENTIATION Dememorization phase marked by reversal of organized structures and functions leading to an undifferentiated state Biotechnology: An Introduction

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BATCH CULTURE A suspension culture in which cells grow in a finite volume of medium and follow a sigmoid pattern of growth. Final cell mass is dependent on the quantity of the limiting nutrient. CONTINUOUS CULTURE A suspension culture continuously supplied with nutrient medium by the inflow of fresh medium. The culture volume is kept constant by draining out the equal amount of used medium ADVENTITIOUS Development of organs (shoot, roots, buds, flowers etc) or embryos (embryo like structure) from unusual points of origin, including callus. Biotechnology: An Introduction

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AXENIC A culture without any foreign or undesired life forms. It may include the purposeful co-cultivation of different types of cells, tissues, or organisms ASEPSIS Without infection or contaminating micro-organisms CELL LINE Cells derived from common ancestral cell that acquire the ability to multiply indefinitely in culture AUTOTROPHIC CULTURES A culture capable of self nourishment Biotechnology: An Introduction

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PROTOPLASTS Single cells with their walls stripped off; may or may not contain active nucleus CYBRIDS A viable cell resulting from fusion of a cytoplasm with a cell; nucleus of one and cytoplasm of both the fusing partners CYTOPLAST Intact cytoplasm remaining following enucleation of a cell SOMACLONAL VARIATION Expression (spontaneous as well as induced) of a phenotypic variation, either genetic or epigenetic in origin displayed by cultured somatic cells in vitro Biotechnology: An Introduction

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CHEMOSTAT An apparatus for maintaining cells (mostly of microbial systems) in the exponential phase of growth over prolonged periods BIOREACTOR A vessel in which a bioprocess takes place BIOCONVERSION A chemical conversion using a biocatalyst ( organism, cell or part there from) IMMOBILIZATION Holding of plant cells so that their metabolic and growth machinary is immobilized by entrappment in polymeric matrices and using such cells for continuous conversion of substrate into products Biotechnology: An Introduction

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SPHEROPLAST A cell from which most of the cell wall has been removed VARIANT A cell line or plant exhibiting a stable phenotypic change that may be genetic (mutant) or epigenetic in nature ACCLIMATIZATION Biological process whereby an organism adapts to a new environment Biotechnology: An Introduction

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CHEMOSTAT CULTURE An open continuous culture in which growth rate and cell density are held constant by a fixed rate of input of growth limiting nutrient(s) TURBIDOSTAT An open continuous culture into which fresh medium flows in response to an increased turbidity. A pre-selected biomass density is uniformly maintained by washing off the excess cells CONTINUOUS CULTURES INSPITE OF THEIR UNIQUE ADVANTAGES ARE NOT FREQUENTLY USED IN LARGE SCALE CULTIVATION BECAUSE OF COSTLY ON-LINE MONITORING Biotechnology: An Introduction

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APPLICATIONS OF PLANT TISSUE CULTURE Rapid propagation of true-to-type plants in limited space and shorter time (MICROPROPAGATION) Production of disease free or disease tolerant plants (MERISTEM CULTURE) As sources of variation (mutants) with better agronomic/productivity traits (SOMACLONAL VARIATION) Production of haploid plants and pure line via anther, pollen and ovule culture (ANDROGENESIS/ GYNOGENESIS) Biotechnology: An Introduction

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APPLICATIONS OF PLANT TISSUE CULTURE Protoplast culture and fusion to create novel plant types with unique cytoplasmic and nuclear combination across evolutionary limits (SOMATIC HYBRIDIZATION) Industrial production of natural products (METABOLIC ENGINEERING) Ex-situ conservation of biodiversity (TISSUE BANKING) Creation of genetically modified plants using genomic approaches (GENETIC ENGINEERING) Molecular markers for linkage mapping and mapping of QTL (Which is a powerful tool for indirect selection for quantitative traits. (RFLPs & RAPDs, Molecular markers) Biotechnology: An Introduction

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GOLDEN RULE S TO SUCCESS IN PTC IN VITRO PLANT TISSUE CULTURE APPROACHES REQUIRE HUGE INFRA-STRUCTURAL INVESTMENTS (COSTLY BUSINESS) THEY SHOULD BE RESOLVED TO ONLY WHEN OTHER OPTIONS ARE EXHAUSTED THEY SHOULD BE USED AS A COMPLIMENTARY INTERPHASE TO HASTEN OR TO ADD NOVELTY TO THE TRADITIONAL METHODS OF PLANT IMPROVEMENT STAY AWAY FROM NATURE FOR MINIMUM TIME AND GET BACK TO NATURE AS SOON AS POSSIBLE Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Biotechnology: An Introduction

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Hardening of in-vitro raised plants in the glass house 3 months after transplantation Biotechnology: An Introduction

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Biotechnology: An Introduction Germination of atrificial Seeds

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MICRO-CLONING IN CROPS Biotechnology: An Introduction

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TISSUE BANKING Biotechnology: An Introduction

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Biotechnology: An Introduction TISSUE BANKING

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Biotechnology: An Introduction Cryopreservation

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Biotechnology: An Introduction Thank You All !

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