Microorganisms Present on Leaf | Biotechnology

In this article we will discuss about the isolation and collection of microorganisms present on the leaf.

Isolation of Microorganisms on Leaf:

Leaf Impression Method:

Principle:

By this method almost all species of organ­isms can be brought into culture.

Requirements:

1. Agar plates—Bacterial, fungal and Actinomycete media.

2. Undamaged healthy mature leaves.

3. Graph paper.

Procedure:

1. Placing the upper surface of the leaf touch­ing the agar plate, gently press and remove. Similarly on another area of agar plate or on a different plate, place the lower surface of the leaf and press gently and remove the leaf.

2. After incubation at room temperature (28°-30°C) for 72-96 hours fungi and actinomycetes and within 24—48 hours bacteria appear. Count number of colo­nies and subculture them.

3. Trace the leaf on a graph paper. Find out the square centimetre area of the leaf and calculate the number of microorganisms per square centimetre.

4. Identify the organisms, subculture on slants and use for further studies.

Collection of Leaf Leachates:

Principle:

Mature leaves have maximum quantity and quality of substances leached on the surface which can be washed. Washings are concen­trated and analysed.

Requirements:

1. Actively growing leaves.

2. Distilled water.

3. Shaker.

4. Water bath.

5. Paper Chromatographic equipment’s.

6. Flasks.

Procedure:

1. Collect clean actively growing leaves from the middle portion of the plant by clipping them without petiole.

2. Weigh one gram of leaves and place them in 99 ml distilled water in 250 ml flasks.

3. Shake at 1200 rpm for half an hour.

4. Remove the leaves and evaporate the solution on a water bath or under vacuum to obtain one to two ml of leachate.

5. Use this for analysis. Chromatography is one method of identifying sugars, amino acids, organic acids, etc.