Filamentous Fungi

Introduction

• Fungi are heterotrophic eukaryotic organisms.

• They are abundant in surface soils and important for their role in
nutrient cycling and decomposition of organic matter and organic
contaminants.
• In this lab we will isolate, observe, and quantify filamentous soil fungi
using dilution and plating techniques.
counted after they have formed macroscopic fungal colonies
Steps
Dilution of the soil is made by suspending a given
amount of soil in a solution (often deionized water).

Transferring aliquots of the suspensions to fresh

solution until the suspension is diluted sufficiently

Inoculation on several replicate agar plates

Counted after they have formed macroscopic fungal

colonies
Dilution
• 10-fold dilution series is performed.
• 10 g of moist soil is added to 95 ml of deionized water and shaken
well to disperse the organisms.
• 10 g of soil occupies approximately 5 ml. we have 10 g of soil in 100
ml total volume, thereby forming a 1 : 10 w/v dilution.
• 1.0 ml of suspension is removed from the bottle and added to a tube
containing 9.0 ml of the same solution.
• The dilution series is continued to the highest desired dilution.
Serial dilution ( 10 fold)
Plating dilution
Pour plates are utilized for the plating procedure.
Dilution and plating calculation
10 g soil → 95 ml saline (solution A) 10-1 (weight/volume)
1 ml solution A → 9 ml saline (solution B) 10-2 (volume/volume)
1 ml solution B → 9 ml saline (solution C) 10-3 (volume/volume)
1 ml solution C → 9 ml saline (solution D) 10-4 (volume/volume)

1 ml of solution C is pour plated onto an appropriate medium and

results in 200 bacterial colonies.
Number of CFU = dilution factor x number of colonies/amount plated

104 x 200 = 200 x 106 CFU/ g moist soil

Result