1. UNIVERSITY OF NAIROBI DEPARTMENT OF PLANT SCIENCE AND CROP PROTECTION PROJECT PROPOSAL PRESENTATION UNIVERSITY OF NAIROBI DEPARTMENT OF PLANT SCIENCE AND CROP PROTECTION PROJECT PROPOSAL PRESENTATION TITLE: EFFECTS OF DIFFERENT GELLING AGENTS IN TISSUE CULTURE OF CITRUS NAME: LUCAS OMONDI ADM. NUMBER: A22/0076/2009 SUPERVISED BY: DR. T. MAGOMERE

2. General objective of the project General objective of the project This project was designed to examine the effects of different media supports, brands gelling agents (agar and phytagel) in the tissue culture of citrus. Specific objectives To study the best levels of both the agents for making media. To compare the growth rates of citrus under mediums containing the gels.

3. Problem statement Agar is the most frequently used solidifier in plant tissue culture media with characteristics like high gel clarity, stability and resistance to digestion by plant enzymes. However, reports on its adverse effects i.e. inhibition of growth, presence of impurities and impartment have been published ( Romberger & Tabor, 1971; Debergh et al. 1981; Debergh, 1983)

4. Project justification It was therefore probable to compare agar with another substitute (phytagel) to see whether the problems stated are manifested in both the gels or not. The rate of productivity of citrus plantlets can highly increased if an alternative gel with optimal properties required by the plants can be used.

5. MATERIALS AND METHODS Plant material (citrus) - Young citrus seeds obtained from 3-4 weeks old fruits. These were purchased from the local market then seeds extracted. Culture medium- Murashige & Skoog`s(1962) culture medium with each level of gel having 150ml of the medium. Agar and phytagel powders.

6. Materials cont. Working laminar flow hood Sterile forceps and sterile petri dishes Sterile distilled water Tween 20 Sterillant e.g. 1% jik 70% ethanol Clean paper towels Burner

7. Plant material preparation The young citrus fruits were carefully cut horizontally to extract the seeds. Seeds were washed with running tap water and quickly dipped in 70% ethanol. The seeds were then soaked in 1% jik for 10 minutes with two drops of tween 20 (surfactant) Seeds were then rinsed with sterile distilled water three times and then placed in petri dish.

8. Addition of gel and culturing The experiment had six levels of both agar and phytagel with each replicated five times. For agar, 3.0, 4.0, 6.0, 8.0, 9.0 and 10.0g/l were used, while in phytagel, 1.0, 1.5, 2.0, 2.5, 3.0 and 4.0g/l were used.

9. Culturing cont. Three seeds were then placed in each baby jar in a semi submerged position using sterile forceps in the lamina flow. The jars were then transferred to the growth room and incubated at 25 0 C for 16/8 photoperiod, at low light intensity.

10. Parameters to be measured Rate of germination  Earliest germinations  Moderately early germinations  Latest germinations Rate of growth  Shoot growth  Root growth General comparison of the gels

11. Budget and work plan Purchase of the 45 young citrus fruit at a total cost of Ksh.200 Every other material was provided by the department at the tissue culture laboratory including the gelling agents.

12. Work plan Culturing done on 12/2/2013 in the morning. First record of germination done after 4 weeks Second record of germination taken after 6 weeks. Record of seeds that have failed to germinate after 8 weeks. Comparison of growth rate after 8 weeks.

13. References Gamborg, O.L., Murashige, T., Thorpe,T.A., and I.K, 1976 Plant tissue culture media. In vitro 12:473-478 Kochba, J., Spiegel-Roy,P., and H.Safran,1972. Adventive plant from ovules and nucelli in citrus plants. Kimani, W., and Jane M. Mbaratha, 1990. Plant tissue culture for propagation and production of disease-free plants Thank you