Materials and methods

Plant material
This study was conducted in the laboratory of the  Department of  Plant Protection, Faculty of  Agriculture, Universiti Putra Malaysia.
The leaves of Ficus deltoidea were collected from Taman Herba Unit Taman Pertanian Universiti, Universiti Putra Malaysia. About one kilogram fresh plant materials were washed under running tap water, oven dry for four days and finally stored until further use 

Microorganisms 

The fungal stock cultures were obtained from the Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia. Two fungal species were used in this experiment which was Ganoderma boninense, and Rhizoctonia solani. R. solani was maintained on Potato Dextrose Agar (PDA) while G.boninense was cultured on Malt Extract Agar (MEA) and stored in the culture chamber under laboratory conditions.

Preparation of plant extract 

The leaves of F. deltoidea were thoroughly washed, oven dried and crushed by using a commercial blender
(Retsch Model SK 100).

Leaf powder of 50g was soaked in 300 ml of methanol in a 500ml conical flask and loaded onto an orbital shaker at a speed of 120 rpm for 24 h. The mixture was filtered using Whatman No-l filter
paper and the filtrate concentrated using rotary evaporator
(Buchi Model R215W).

Dried extract was collected in a conical flask covered with aluminiums foil, sealed with parafilm and stored at 4°C. 

Screening of antifungal activity

By using poison agar technique that involved the cultivation of pathogen plugs on a culture medium containing F. deltoidea leaf extract, and then the percent inhibition of diameter growth (PIDG) was determined by measuring the diameter of mycelial growth daily until a week for G.boninense and five days for R.solani, F.deltoidea leaf extract with different concentrations as treatments were incorporated and mixed well with respective culture medium and poured into culture plates. 

In this experiment four different concentrations of leaf extract were used which were 5%, 10%, 15% and 20% and compared with control (0%). Stock solution was prepared by diluting the extract with methanol. The obtained crude extract was added to 100 ml methanol and left it for 24 h in the laminar air flow. To make up other concentrations, for example, 5%, 1 m1 of stock solution was taken and added into a Petri dish and was added with 19 ml agar solution.

For each treatment, three replicates were done. Agar plugs of G. boninense and R. solani were placed at the center of Petri dishes containing MEA and PDA respectively, with extracts at the defined concentrations.

The formula used to calculate the percent of inhibition was:

{[diameter of mycelial growth of fungal pathogen in control plate- Diameter of mycelial growth of fungal pathogen in treatment plate]  /Diameter of mycelial growth of funga1 pathogen in control plate} x 100

Experimental design and statistical analysis

Recorded data was analyzed with SAS Software (SAS Institute, North Carolina, State University, Version 9.1, 2004). The effects and the interactions between treatments and phatogens were tested by ANOVA using CRD (Completely Randomized Design) where there were five treatments with three replications for each treatment and the means of significant results were compared using Least Significance Difference (LSD) at 5% probability level.