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1. Soil Sampling - Coggle Diagram

- - - - Significant differences accepted at p<0.05, p<0.01 , p<0.001*
        
        Principal component analysis (PCA) was applied to examine PLFA community structure on the different samples
        
        Statistics calculated using the Statistica 6.0 technology.
    - - 10g soil sample diluted in 90ml of sterile water
        
        10-fold serial dilutions are made
        
        CFU are determined according to different methods based on bacterial, fungi and actinomycetes origin
        
        CFU counted after 5 days for bacteria / 7 days for fungi and actinomycetes at 28 degree Celcius
        
        PLFA analysis performed using 2 different methods for bacterial and fungal respectively
        
        Ratio of GN/GP is used as indicator for changes in the relative abundance.
        
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  - - - The concentrations then added to the treated pots (Control pots : only distilled water)
        
        Each is triplicated : :
        
        Pots sealed
        
        Placed in environment chamber at 25 degree Celcius air temperature and 50% humidity in the dark
        
        Aeration once a day for 1 hour. Soil moisture maintained at 60% water-holding capacity with deionized water.
        
        Pots taken out randomly from environment chamber after 1,8,15,22 and 29 days ( CFU analysis) and 29 days (microbial community analysis PLFA)
- - - - Duplicate composite samples taken with 10 subsamples, mixed, packed, labelled
        
        Transported to the laboratory
        
        Samples mixed, sieved with 2mm mesh to remove plant tissue
        
        Samples pre-incubated in the dark at 25 degree Celcius, 50% humidity for 14 days
        
        Addition of 2,4-D butyl ester