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Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM PLANT PROPAGATION (AN OVERVIEW)

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Presentation on theme: "Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM PLANT PROPAGATION (AN OVERVIEW)"— Presentation transcript:

1 Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM PLANT PROPAGATION (AN OVERVIEW)

2 Kodiswaran Kandasamy © 2006 PLANT PROPAGATION - conventional - tissue culture

3 Kodiswaran Kandasamy © Cuttings (leaf, stem, root) 2. Layering (stem) 3. Marcotting 4. Grafting 5. Seed CONVENTIONAL PROPAGATION

4 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

5 Kodiswaran Kandasamy © 2006

6 plant tissue culture some basics

7 Kodiswaran Kandasamy © 2006 Description Growing surface sterilised plant tissue in a nutrient medium under aseptic conditions in a controlled environment

8 Kodiswaran Kandasamy © 2006 Hormonal control of growth AUXIN Low High IBA NAA IAA CYTOKININ High Low BAP Kinetin 2iP Zeatin shoots roots

9 Kodiswaran Kandasamy © 2006 Effect of IAA and Kinetin on Proliferation and morphogenesis: Auxin Cytokinin (mg/l)

10 Kodiswaran Kandasamy © 2006 MICROPROPAGATION (commercial)

11 Kodiswaran Kandasamy © 2006 Laboratory design Defined culture procedures Scheduling & Stock keeping Calculations Human resource Key factors

12 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

13 Kodiswaran Kandasamy © 2006 Micropropagation cycle Laboratory Greenhouse

14 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

15 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

16 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

17 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

18 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

19 Kodiswaran Kandasamy © 2006 frim at a brief

20 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

21 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

22 Kodiswaran Kandasamy © 2006 Introduction R&D Services Staff Products Contacts TISSUE CULTURE LABORATORY (MAKMAL KULTUR TISU)

23 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

24 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

25 Kodiswaran Kandasamy © 2006 Shorea leprosula

26 Kodiswaran Kandasamy © 2006

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28 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

29 Kodiswaran Kandasamy © 2006 MATURE PODS IMMATURE PODS

30 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

31 Kodiswaran Kandasamy © 2006 for Somatic Embryogenesis for Micropropagation Explant source Shoot formationRegeneration Culture initiation Shoot multiplication Callus initiationCallus proliferation Neobalanocarpus heimii (Chengal)

32 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)

33 Kodiswaran Kandasamy © 2006 Tongkat Ali (somatic embryogenesis)

34 Kodiswaran Kandasamy © 2006 Mangrove species

35 Kodiswaran Kandasamy © 2006

36 Weaning Chamber Patent Pending: (# PI )

37 Kodiswaran Kandasamy © 2006 Teak (Tectona grandis)

38 Kodiswaran Kandasamy © 2006 Acacia (Acacia hybrids)

39 Kodiswaran Kandasamy © 2006 Jelutong (Dyera costulata)

40 Kodiswaran Kandasamy © 2006 Sesenduk (Endospermum diadenum)

41 Kodiswaran Kandasamy © 2006 Karas (Aquilaria malaccensis)

42 Kodiswaran Kandasamy © 2006 Sungkai (Peronema canescens)

43 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)

44 Kodiswaran Kandasamy © 2006 other experiences a quick recap

45 Kodiswaran Kandasamy © 2006

46 Somatic embryogenesis

47 Kodiswaran Kandasamy © 2006 Suspension cell culture

48 Kodiswaran Kandasamy © 2006 Artificial seed production

49 Kodiswaran Kandasamy © 2006 Anther & pollen culture

50 Kodiswaran Kandasamy © 2006 Protoplast culture

51 Kodiswaran Kandasamy © 2006 Protoplast fusion

52 Kodiswaran Kandasamy © 2006 Fungal interaction studies

53 Kodiswaran Kandasamy © 2006 Associated problems

54 Kodiswaran Kandasamy © 2006 THANK YOU

55 Kodiswaran Kandasamy © 2006 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM

56 Kodiswaran Kandasamy © Disease tolerance 2. Climatic tolerance 3. Value added products 4. Higher/improved yield 5. Change characteristics Cost & time Kodiswaran Kandasamy Genetic modification

57 Kodiswaran Kandasamy © 2006

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59 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?

60 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

61 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

62 Kodiswaran Kandasamy © Time: 15 years 2. Cost: Ca RM15 million 3. Screen: 10,000 compounds 4. Chance: 1 drug Drugs from microbes

63 Kodiswaran Kandasamy © Polyploidization 2. Protoplast fusion 3. Haploid plants 4. Artificial seeds Miscellaneous tools


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