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Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM PLANT PROPAGATION (AN OVERVIEW)
Kodiswaran Kandasamy © 2006 PLANT PROPAGATION - conventional - tissue culture
Kodiswaran Kandasamy © Cuttings (leaf, stem, root) 2. Layering (stem) 3. Marcotting 4. Grafting 5. Seed CONVENTIONAL PROPAGATION
Kodiswaran Kandasamy © Micropropagation 2. Somatic embryogenesis 3. Suspension cell Culture 4. Protoplast fusion 5. Micrografting 6. Polyploidisation 7. Haploid Cell Culture 8. Mutation Breeding 9. Artificial Seeds TISSUE CULTURE
Kodiswaran Kandasamy © 2006
plant tissue culture some basics
Kodiswaran Kandasamy © 2006 Description Growing surface sterilised plant tissue in a nutrient medium under aseptic conditions in a controlled environment
Kodiswaran Kandasamy © 2006 Hormonal control of growth AUXIN Low High IBA NAA IAA CYTOKININ High Low BAP Kinetin 2iP Zeatin shoots roots
Kodiswaran Kandasamy © 2006 Effect of IAA and Kinetin on Proliferation and morphogenesis: Auxin Cytokinin (mg/l)
Kodiswaran Kandasamy © 2006 MICROPROPAGATION (commercial)
Kodiswaran Kandasamy © 2006 Laboratory design Defined culture procedures Scheduling & Stock keeping Calculations Human resource Key factors
Kodiswaran Kandasamy © 2006 Appropriate size of each section Media Prep & Washing Autoclave & Media room Transfer Room Growth Rooms Work Flow (Layout) From non-clean to clean Utilities & Specification Clean water Reliable power supply Floor & Wall finish Equipment layout Laboratory design
Kodiswaran Kandasamy © 2006 Micropropagation cycle Laboratory Greenhouse
Kodiswaran Kandasamy © 2006 Define Explant Type & Size Define Developmental Stages Define Media for Each Stage Define Growth Room Conditions Define Number of Explant/Vessel Define Sub-Culture Interval (cycle) A Complete Step-by-Step guide Shoot-tipNodal explant Plantlets ready for next sub-culture Culture procedures
Kodiswaran Kandasamy © 2006 Continuous production Multiplication cycle - numbers Rooting Stages - numbers Ready for Sale (ex-lab) - numbers Allocation of resources Equipment Manpower Raw material Stock keeping (database) Production record & estimation Plan production to meet targets (demand) Tracking & able to trace stock Scheduling & stock keeping
Kodiswaran Kandasamy © 2006 Production estimations Input requirement & cost Raw material Labour Production efficiency Cost estimations Budget estimation Business plan Cash flow (P&L) Marketing Delivery schedule Calculations
Kodiswaran Kandasamy © 2006 Smooth running of production line Skilled & semi-skilled Strict job specification Production Manager Production Supervisor Operators Support staff Monitor & evaluate performance Healthy and safe workplace Human resource
Kodiswaran Kandasamy © 2006 Planning production Y = No. of plantlets produced A = No. of shoots produced at each subculture B = No. of initial explants available n = No. of subculture passages F 1 = Proportion of usable shoots after Stage II F 2 = Proportion of usable shoots after Stage III F 3 = Proportion of usable shoots after Stage IV Y = A n x B x F 1 x F 2 x F 3
Kodiswaran Kandasamy © 2006 frim at a brief
Kodiswaran Kandasamy © 2006 Genetic resources Forests are the main home of Earth's species. While covering only 6% of the planet's land surface, tropical forests contain at least 50%, possibly 70% and conceivably 90% of all species. ½ km 2 of Malaysia's forests can feature as many tree and shrub species as the whole of the United States and Canada, while a single bush in Peruvian Amazonia has revealed as many ant species as in the British Isles. Norman Myers
Kodiswaran Kandasamy © 2006 Growing more crops and trees per acre leaves more land for Nature We cannot choose between malnourished children & saving endangered wild species Without higher yields, peasant farmers will destroy the wildlands and species to keep their children from starving Sustainably higher yields of crops and trees are the only visible way to safe both Dr. Norman Borlaug
Kodiswaran Kandasamy © 2006 Introduction R&D Services Staff Products Contacts TISSUE CULTURE LABORATORY (MAKMAL KULTUR TISU)
Kodiswaran Kandasamy © 2006 Facilities & Infrastructure We are fully equipped to undertake a complete range of in vitro studies, i.e. micropropagation, suspension cell culture, somatic embryogenesis, etc. Media Preparation Room Weaning Chamber Transfer Room Growth Rooms Culture Racks Holding Area Growing AreaMisting Area
Kodiswaran Kandasamy © 2006 Research & Development Development of suitable, economically viable mass propagation protocol for various forest species. Selected ParentExplant Material Culture InitiationMultiplicationWeaned Plantlet Somatic EmbryosS.E. germinationShorea (Rita System)Embryogenic callus Cell suspension
Kodiswaran Kandasamy © 2006 Shorea leprosula
Kodiswaran Kandasamy © 2006
Wood density (AB6 x MB20) Fiber Length (AB3 x MB22 Harvest (pods/seeds) Transfer to laboratory Surface sterilisation In vitro germination Confirmation (DNA/Isozyme) MultiplicationTransplant Hybrids Non-hybrids Discarded Clonal Trial Acacia hybrids
Kodiswaran Kandasamy © 2006 MATURE PODS IMMATURE PODS
Kodiswaran Kandasamy © 2006 Crosses > 70,000 Seed Pods > 380 Seeds > 2,000 Seedlings (ca 70%)> 1,400 Selected clones = 400 Plantlets (for clonal trial) = 20,000 In vitro germination & plant production
Kodiswaran Kandasamy © 2006 for Somatic Embryogenesis for Micropropagation Explant source Shoot formationRegeneration Culture initiation Shoot multiplication Callus initiationCallus proliferation Neobalanocarpus heimii (Chengal)
Kodiswaran Kandasamy © 2006 Culture Establishment from Seed Explant ex vitro germination in vitro germination Seed explants Seed dissected Germination (in vitro) Seedling (in vitro) Germination (ex vitro)Seeding (ex vitro)
Kodiswaran Kandasamy © 2006 Tongkat Ali (somatic embryogenesis)
Kodiswaran Kandasamy © 2006 Mangrove species
Kodiswaran Kandasamy © 2006
Weaning Chamber Patent Pending: (# PI )
Kodiswaran Kandasamy © 2006 Teak (Tectona grandis)
Kodiswaran Kandasamy © 2006 Acacia (Acacia hybrids)
Kodiswaran Kandasamy © 2006 Jelutong (Dyera costulata)
Kodiswaran Kandasamy © 2006 Sesenduk (Endospermum diadenum)
Kodiswaran Kandasamy © 2006 Karas (Aquilaria malaccensis)
Kodiswaran Kandasamy © 2006 Sungkai (Peronema canescens)
Kodiswaran Kandasamy © 2006 MAIN ACTIVITY FLOW-CHART Sample Receive d Reject sample (Sample again) Order (received) Record informatio n PROTOCOL DEVELOPMENT Sterilisation Method R & D Culture Initiation Multiplicatio n (culture) Rooting (of shoots) Weanin g No Delivered ( to client) Prepare for productio n Shoot multiplicatio n Rooting of shoots Sales (ex-lab) Weaning (ex- nursery)
Kodiswaran Kandasamy © 2006 other experiences a quick recap
Kodiswaran Kandasamy © 2006
Kodiswaran Kandasamy © 2006 Suspension cell culture
Kodiswaran Kandasamy © 2006 Artificial seed production
Kodiswaran Kandasamy © 2006 Anther & pollen culture
Kodiswaran Kandasamy © 2006 Protoplast culture
Kodiswaran Kandasamy © 2006 Protoplast fusion
Kodiswaran Kandasamy © 2006 Fungal interaction studies
Kodiswaran Kandasamy © 2006 Associated problems
Kodiswaran Kandasamy © 2006 THANK YOU
Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM
Kodiswaran Kandasamy © Disease tolerance 2. Climatic tolerance 3. Value added products 4. Higher/improved yield 5. Change characteristics Cost & time Kodiswaran Kandasamy Genetic modification
Kodiswaran Kandasamy © 2006
1. Look for the gene (s) & remove it 2. Clone it & place it in the desired plant 3. Ensure it functions well & does not disrupt other functions 4. Produce many copies of the plant What it means?
Kodiswaran Kandasamy © 2006 Plant Production Processing & formulating Marketing Hydroponic farming for clean & vigorous growth & quick harvest Processing plant Freeze drying Grinding Product formulation MARKET R&D Pegaga grown via Tissue Culture for mass volume production 1. Health food supplement 2. Natural medicine 3. Drugs Cost & time Extraction, Purification & Standardisation Pharmaceuticals/Nutraceuticals
Kodiswaran Kandasamy © Plantibodies (diagnostics & vaccines) 2. Therapeutic proteins (e.g. Interferon, growth hormones) 3. High value enzymes for industry 4. Plant defense (e.g. Virus resistant plants) Cost & time Green pharmacy
Kodiswaran Kandasamy © Time: 15 years 2. Cost: Ca RM15 million 3. Screen: 10,000 compounds 4. Chance: 1 drug Drugs from microbes
Kodiswaran Kandasamy © Polyploidization 2. Protoplast fusion 3. Haploid plants 4. Artificial seeds Miscellaneous tools
Basic Principles & Protocol in Plant Tissue Culture Chapter 9.
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Structure, function and growth of prokaryote and eukaryote cells (iii) Differentiation of cells into tissues and organs. (iv) Cell and tissue culture –
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Organogenesis in Peanut Research Experience for Undergraduates Food Biotechnology Laboratory Alabama A&M University Antonio Brazelton 7/3/08.
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Plant Tissue Culture. Definition: The term plant tissue culture is described in vitro and aseptic cultivation of any plant part on nutrient medium.
Lecture 8 Plant Tissue Culture & Applications. What is it? Tissue culture is the term used for “the process of growing cells artificially in the laboratory”
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Lec # Application of plant tissue culture & Biofertilisers Dr. Shah Rukh Abbas
India is one of the major banana exporting countries. Banana crop is an important source of income and employment in South and Central India. Market sale.
5 d) Cloning 5.17 describe the process of micropropagation (tissue culture) in which small pieces of plants (explants) are grown in vitro using nutrient.
IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)
Agricultural Biotechnology. How can we obtain better Crops ? 1.selection 2.breeding 3.hybridization 4.cloning 5.grafting 6.radiation mutagenesis 7.chemical.
Reminder - Your Tasks for Monday 1. Home Learning – Assessed Report Using drosophila as an example explain the process of differentiation of cells into.
PRINCIPLES OF CROP PRODUCTION ABT-320 (3 CREDIT HOURS) LECTURE 11 BIOTECHNLOGICAL APPROACHES IN PLANT BREEDING, IN VITRO CULTURE TECHNOLOGY, MICROPROPAGATION,
Naomi Asomani Antwi Matilda Ntowah Bissah May, 2016.
AGRICULTURAL BIOTECHNOLOGY. Stages of Biotechnology Development Ancient biotechnology early history as related to food and shelter; Includes domestication.
PLANT PROPAGATION Propagation The multiplication of a kind or species. Reproduction of a species.
Plant Tissue Culture A rice plant growing in nutrient rich agar Tissue Culture A.K.A Micropropagation In vitro Culture faperta.ugm.ac.id/newbie/download/pak_tar/Plantbiotechnology/TissuC.
Tissue Culture Unit one Cell and Molecular Biology Advanced Higher Biology.
Chrysanthemum ‘Golden Surfer’ protoplast regeneration Tom Eeckhaut, Silvia Bruznican and Johan Van Huylenbroeck Institute for Agricultural and Fisheries.
Micropropagation “… the art and science of multiplying plants in vitro.”
Biotechniques. Tissue Culture (micro-propagation) A method of rapidly cloning desired plants [few weeks]. Plant cells are totipotent (can grow into whole.
2.5 Asexual reproduction in plants. Propagation Asexual reproduction: Asexual reproduction is a mode of reproduction by which offspring arise from a.
Biotechnology Lecture 1 Applications on Plants. Tissue culture or In vitro culture Plant tissue culture: tissue culture means the culturing of pieces,
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Lec# 11 Plant tissue culture, GE Plants and applications Dr. Shah Rukh Abbas
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CHAPTER 38 PLANT REPRODUCTION AND BIOTECHNOLOGY Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Section B: Asexual Reproduction.
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Successful regeneration of A. paraguariensis was achieved. Direct multiple shoot formed on cotyledon explants within five days of culture incubation. Thidiazuron.
An Ethical Debate. Humans have been breeding farm animals for thousands of years Selective breeding is done to get the traits a person wants.
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