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Presentation on theme: "PLANT PROPAGATION (AN OVERVIEW) Kodiswaran Kandasamy"— Presentation transcript:

Tissue Culture Unit Forest Biotechnology Division FRIM

2 PLANT PROPAGATION - conventional - tissue culture

1. Cuttings (leaf, stem, root) 2. Layering (stem) 3. Marcotting 4. Grafting 5. Seed

4 TISSUE CULTURE 1. 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


6 plant tissue culture some basics

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

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

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

10 MICROPROPAGATION (commercial)

11 Key factors Laboratory design Defined culture procedures Scheduling & Stock keeping Calculations Human resource

12 Laboratory design 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

13 Micropropagation cycle
Laboratory Greenhouse

14 Culture procedures 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 Nodal explant Shoot-tip Plantlets ready for next sub-culture

15 Scheduling & stock keeping
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

16 Calculations Production estimations Input requirement & cost
Raw material Labour Production efficiency Cost estimations Budget estimation Business plan Cash flow (P&L) Marketing Delivery schedule

17 Human resource 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

18 Planning production Y = An x B x F1 x F2 x F3
Y = No. of plantlets produced A = No. of shoots produced at each subculture B = No. of initial explants available n = No. of subculture passages F1 = Proportion of usable shoots after Stage II F2 = Proportion of usable shoots after Stage III F3 = Proportion of usable shoots after Stage IV Y = An x B x F1 x F2 x F3

19 frim at a brief

20 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 km2 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 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

Introduction R&D Services Staff Products Contacts TISSUE CULTURE LABORATORY (MAKMAL KULTUR TISU)

23 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 Area Misting Area

24 Research & Development
Development of suitable, economically viable mass propagation protocol for various forest species. Selected Parent Explant Material Culture Initiation Multiplication Weaned Plantlet Somatic Embryos S.E. germination Shorea (Rita System) Embryogenic callus Cell suspension

25 Shorea leprosula



28 Acacia hybrids Wood density (AB6 x MB20) Harvest (pods/seeds)
Transfer to laboratory Surface sterilisation Fiber Length (AB3 x MB22 Confirmation (DNA/Isozyme) In vitro germination Non-hybrids Discarded Hybrids Clonal Trial Transplant Multiplication


30 In vitro germination & plant production
Crosses > 70,000 Seed Pods > Seeds > 2,000 Seedlings (ca 70%) > 1,400 Selected clones = Plantlets (for clonal trial) = 20,000

31 for Somatic Embryogenesis
Neobalanocarpus heimii (Chengal) Culture initiation Explant source Regeneration Shoot formation Callus proliferation Callus initiation Shoot multiplication for Somatic Embryogenesis for Micropropagation

32 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 Tongkat Ali (somatic embryogenesis)

34 Mangrove species


36 Weaning Chamber Patent Pending: (# PI )

37 Teak (Tectona grandis)

38 Acacia (Acacia hybrids)

39 Jelutong (Dyera costulata)

40 Sesenduk (Endospermum diadenum)

41 Karas (Aquilaria malaccensis)

42 Sungkai (Peronema canescens)

43 MAIN ACTIVITY FLOW-CHART Prepare for production
Sample Received MAIN ACTIVITY FLOW-CHART Reject sample (Sample again) PROTOCOL DEVELOPMENT Record information R & D No Sterilisation Method Prepare for production Order (received) No Culture Initiation Shoot multiplication No Multiplication (culture) Sales (ex-lab) Rooting of shoots No Rooting (of shoots) Weaning (ex-nursery) Delivered ( to client) No Weaning

44 other experiences a quick recap


46 Somatic embryogenesis

47 Suspension cell culture

48 Artificial seed production

49 Anther & pollen culture

50 Protoplast culture

51 Protoplast fusion

52 Fungal interaction studies

53 Associated problems


55 Kodiswaran Kandasamy Tissue Culture Unit Forest Biotechnology Division FRIM

56 Genetic modification 1. Disease tolerance 2. Climatic tolerance
3. Value added products 4. Higher/improved yield 5. Change characteristics Cost & time Kodiswaran Kandasamy



59 What it means? 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

60 Pharmaceuticals/Nutraceuticals
Processing & formulating Plant Production Marketing R&D Processing plant Hydroponic farming for clean & vigorous growth & quick harvest Pegaga grown via Tissue Culture for mass volume production Extraction, Purification & Standardisation Freeze drying Product formulation Grinding 1. Health food supplement 2. Natural medicine 3. Drugs MARKET Cost & time

61 Green pharmacy 1. 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

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

63 Miscellaneous tools 1. Polyploidization 2. Protoplast fusion
3. Haploid plants 4. Artificial seeds

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