THE MORPHOLOGICAL, PHYSIOLOGICAL AND CHEMICAL STUDIES OF SELECTED MEDICINAL PLANT (Catharanthus roseus) THROUGH

MYCORRHIZAL FUNGI TREATMENT

Farah Wahida Ayob Feb 22, 2013

OBJECTIVES
General objective :

To study the molecular DNA identification of mycorrhizal fungi and the effectiveness of mycorrhizal fungi and fertilizer usage on the morphological, physiological and chemical properties of Catharanthus roseus.
Specific objectives :

1.To identify the mycorrhizal association through the molecular DNA indentification and microscopic method. 2.To examine the effects of mycorrhizal fungi inoculation and fertilizer usage on the morphological, physiological and chemical properties of Catharanthus roseus.

3.To compare the effects of different inoculation technique of mycorrhizal fungi on the productivity of Catharanthus roseus.

HYPOTHESIS

From the research, we expect that

(i) the inoculation of mycorrhizal fungi on Catharanthus

roseus will increase morphological, physiological and chemical properties of the plant.
(ii) the usage of fertilizer will decrease due to the boosting

nutrient uptake by host plant which had the mycorrhizal association.

(iii) the growth of Catharanthus roseus is much more

better through liquid inoculation technique compared to the solid inoculation.

METHODOLOGY 1. Collection of Mycorrhizal Fungi Root Sampling Root Clearing and Colouring Technique Microscopy - Light microscopy
- Scanning electron microscopy (SEM) - Transmission electron microscopy (TEM)

2. Molecular DNA identification 3. Experimental Design 4. Inoculation technique (liquid / solid inoculum) 5. Plant Growth Measurement
morphological - (plant height, root length, root dry weight, root fresh weight, stem diameter, number of leaves and ratio of root colonization) physiological (the content of nitrogen, phosphorus and pottasium) chemical (alkaloid vincristine, vinblastine, ajmalicine) 6. Harvesting

and Staining of Root Samples 7. Statistical Analysis

METHODOLOGY cont.
Experimental Design
Mycorrhizal inoculated Control Sterilized soil Fertilizer -

Treatment 1
Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6 Treatment 7

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+ + + -

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+ + + -

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METHODOLOGY cont.
Catharanthus roseus

Seed

Seedling

Liquid inoculation
7 treatmant + control

Solid inoculation
7 treatmant + control

Liquid inoculation
7 treatmant + control

Solid inoculation
7 treatmant + control

FLOW CHART
STAGE 1 Identification of the mycorrhizal fungi in the host plants root *Inoculate *Microscopy (LM, SEM, TEM) *DNA test (PCR) STAGE 2 The preparation of inoculums in the culture media

STAGE 3 7 experiments and control seedling (experimental design) Liquid and solid inoculation

DATA COLLECTION *plant growth measurement *Microscopy (LM, SEM, TEM) *DNA test (PCR)

GANTT CHART
Methods Month Proposal 9 10 2012 11 12 1 2 3 4 5 6 2013 7 8 9 10 11 12 1 2 3 4 5 6 2014 7 8 9 10 11 12

Presentation
Literature review

Experiment Stage 1
(AMF isolation)

Stage 2
(Inoculums preparation)

Stage 3
(Experimental design )

Data collection

Progress report

Data analysis Writing

MILESTONE
Num Date Description

1 2 3

March 2013 May 2013 October 2013

Complete isolation and identification of arbuscular mycorrhizal fungi Present proposal defense (Seminar 1) Complete experiment on seedling (Mycorrhizal Effect on the Growth of Catharanthus roseus Seedling by Using Different InoculationTechnique)

Mei 2014

Complete experiment on seed (Effect of Mycorrhizal Fungi on the Productivity of Catharanthus roseus Seed through Liquid and Solid Inoculation)

5 6

Jun 2014 August 2014

Complete data collection Complete data analysis (Ultrastructural Changes of Catharanthus roseus Root after Treated with Mycorrhizal Fungi.

February 2015

Complete thesis writing.