Isolation of phylloplane microorganism
Phyllosphere term is used for the microflora of leaf surface. Therefore the term phylloplane was suggested for leaf surface environment by last and deighton. Thus, phylloplane contain a large quantity of microorganisms both pathogens and saprophytes. There are several methods used for isolation of phylloplane microorganisms but the most common are leaf impression method and serial dilution plate method.
Isolation of microorganisms by leaf impression method
In this method leaves are gently pressed on the surface of nutrients agar medium in petrified plates. After incubation microorganisms grow are isolated, identified and preserved if necessary.
Requirements
- Plant leaf sample
- Sterilization polythene bags
- Czapek Dox agar plates (for fungi)
- Nutrient agar plates ( for bacteria)
- Bunsen burner
- Marking pencil
Procedure
- collect leaves of different age groups from the desired plant, put in sterilization polythene bags and bring them to the laboratory.
- Press aseptically first the ventral surface of a leaf and then dorsal surface on the surface of agar plates.
- Incubate the Nutrient agar plates for 24-48 hours and Czapek Dox agar plates for 5-7 days at 25 C in inverted position.
Observation
different microorganisms grow on agar plates. Count their number per plate to measure microbial population.
Reference
Dr. R. C. Dubey – Practical Microbiology
