What is Plant tissue culture?

1. Use of Low Cost Tissue Culture Materials for the Initiation and multiplication of Plants a case of Bananas, Cassava and Sweet potatoes Gitonga N. Mburugu Meru University College of Science and Technology

2. What is Plant tissue culture? • Refers to growing and multiplication of cells, tissues and organs on; defined media under aseptic and controlled environmental conditions

3. Limitations of conventional tissue culture • High cost of equipment • High cost of nutrient media and sterilizing agents • High cost of facilities • Lack or shortage of trained personnel • Lack of systems for marketing/ delivering tissue culture products

4. Solution: • Develop low cost tissue culture technology in terms of • Equipment • Chemicals • Apparatus • Protocols • Personnel/skills required/programs

5. What Low Cost Tissue Culture entails • Use of alternative and where possible locally available equipment and resources to • reduce the unit cost of tissue culture products without compromising the quality of the plants

6. Points of intervention in TC cost reduction Plant TC has three components which form points of intervention in cost reduction: Chemicals (minerals nutrients, plant growth hormones, vitamins) Equipment (culture containers, autoclave, laminar flow, instruments used for micropropagation, pH meter etc) Structures (media preparation, inoculation, growth and hardening rooms)

7. Phase one (preparation room) A: Equipment ELECTRIC AUTOCLAVE is expensive; long heating time, requires electricity, specialized maintenance, risk of electric shock. PRESSURE COOKER is cheap, efficient, can use any source of heat, easy to maintain, safe Substitution of preparation room equipment with low cost options has been successful

8. Cont’d Costly, delicate pipettes and cylinders Replaced by cheap, hardy syringes

9. Cont’d B: Disinfectants Costly Sodium Hypochlorite Low cost Jik®

10. C: Culture nutrients A: Easygro® fertilizer used in LCM 1 B: Stanesiodisedmicrofood® used as a source of micronutrients in LCM 2 B A Some of the conventional MS salts C

11. Cont’d Conventional vitamins Vitamin B complex tablets

12. Vitamins • NAA (Anatone) was found as effective as the conventional NAA. • GA3 (Tivag) was effective in shoot elongation. • Plantone - a source of BAP- was found effective in shoot induction.. • Pyridoxine and nicotinamide -readily available as tablets

13. Solidifying agent Agar or gerilite • Extraction of Agar from sea weed ??????

14. Phase two (Inoculation room) Conventional hood costly as it requires air filters, air pump, UV light, clear door, stainless steel bench, working light. Low cost hood fabricated using wood and formica

15. Final Phase (Growth room) Conventional electrically powered growth room Low cost growth room under natural light • Most challenging phase

16. Cont’d • We been successful with LCTC for • bananas Gitonga et al 2009) • Cassava ( Kwame 2011) • sweet potato ( Kwame 2011) • Cock-yams (Known in East Africa as Arrow roots) (Kibet in progress) • These are crops largely grown by subsistence farmers in sub-Saharan Africa.

17. B c A E D Low cost propagation of Bananas

18. Cassava plantlets on LCM 1: A: Muchericheri B: KME 1 A B Cassava plantlets on LCM 2: A: Muchericheri B: KME 1 A B

19. Acclimatization of cassava plantlets on Vermiculite in a bucket Cassava plantlets after transfer onto soil

20. Sweet potato plantlets on LCM 1: A: KEMB 36 B: Tainurey A B Sweet potato plantlets on LCM 2: A: KEMB 36 B: Tainurey A B

21. A B A: Sweet potato plantlets acclimatized on a mixture of red soil and rice husks B: Sweet potato plantlets transplanted on soil

22. Kawanda Agricultural Research Institute (KARI, Uganda)

23. Agro-Genetic Technologies Ltd ErostusNsubuga,CEO AGT and Chairman TCBN E & CA

24. AGT is doing booming business supplying Uganda, Rwanda and Congo • It’s currently the leading commercial TC institution in ECA

25. In Kenya Genetic Technologies Ltd leads followed by J.K.U.A.T and Mimea International Ltd JKUAT net house with thousands of banana seedlings ready for sell

26. Milestones: Table 1a: Substitution of macronutrients in basal Murashige and Skooge plant culturing Media Key Kes Kenya shillings CMA - Convectional macronutrients SMA - Substitute Macronutrients

27. Milestones: Table 1b: Substitution of micronutrients in basal Murashige and Skooge plant culturing Media Key Kes Kenya shillings CMI - Convectional macronutrients SMI - Substitute Macronutrients

28. Milestones: Table 1c: Substitution of Vitamins and other supplements in plant culturing Media

29. Milestone: Table 1d: Substitution of growth regulators in Murashige and Skoogeplant culturing Media Table 1d: Substitution of growth regulators in Murashige and Skooge plant culturing Media

30. Milestone Table 1e: Substitution of carbon source in basal Murashige and Skooge plant culturing Media

31. Milestone Table 1f: Substitution of solidifying agents in basal Murashige and Skooge plant Culturing Media

32. Milestone Table 2: Substitution of plant tissue culturing facility equipment and apparatus Key CE&A – Convectional equipment and apparatus SE&A – Substitute equipment and apparatus

33. Acknowledgement • Funding urgency: Initial funding by Rockefeller foundation, Later ASARECA

34. THANK YOU All • Special thanks for the work • Dr Ombori • S. Kiogora • M. Ngugi • S. ndungu • O kwame • S. kibet • S. kasyoka THANK YOU