UNIVERSITY OF NAIROBIUNIVERSITY OF NAIROBIDEPARTMENT OF PLANT SCIENCE AND CROP DEPARTMENT OF PLANT SCIENCE AND CROP

PROTECTION PROTECTION PROJECT PROPOSAL PRESENTATION 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

General objective of the projectGeneral 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 objectivesTo study the best levels of both the

agents for making media.To compare the growth rates of citrus

under mediums containing the gels.

Problem statementProblem 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)

Project justificationProject 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.

MATERIALS AND METHODS 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.

Materials cont.Materials cont.Working laminar flow hoodSterile forceps and sterile petri

dishesSterile distilled waterTween 20Sterillant e.g. 1% jik70% ethanolClean paper towelsBurner

Plant material preparationPlant 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.

Addition of gel and culturingAddition 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.

Culturing cont.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 250C for 16/8 photoperiod, at low light intensity.

Parameters to be measuredParameters to be measured

Rate of germinationEarliest germinations Moderately early germinationsLatest germinations

Rate of growthShoot growth Root growth

General comparison of the gels

Budget and work planBudget 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.

Work planWork 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.

References 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