UNIVERSITI TEKNOLOGI MARA

LABORATORY MANUAL
MIC 316

AGRICULTURAL MICROBIOLOGY

GROUP
GROUP MEMBERS 1)
2)
3)
4)
LECTURER

FACULTY OF APPLIED SCIENCES

UiTM NEGERI SEMBILAN, KUALA PILAH

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 MIC 316
PRACTICAL 1

Observing symptoms and signs

Objective

1. Learn to recognize, define, and characterize symptoms on diseased plant material.

2. Learn to identify signs of pathogens and differentiate signs from symptoms.
3. Practice making careful observations of both symptoms and signs that can aid in
diagnosis of plant problems.

Materials

Samples of plants/plants parts showing symptoms and signs associated with plant
diseases.

Procedures

1. Observe the symptoms and signs that appear on the plant specimens.
2. Examine the following set of pictures. Name the symptom type (e.g., leaf spot, wilt)
that best fits each disease.

Figure 1 Figure 2

2
Figure 3 Figure 4

Figure 5 Figure 6

Figure 7 Figure 8

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 Figure 9 Figure 10

Results

Draw and label the symptoms or signs of the specimens.

Discussions

1. Which group of symptoms the specimen falls in.

2. Do the symptoms tell you what cause the disease. Why?

Questions

1. Define the following terms:

i) canker
ii) necrotroph
iii) biotroph
iv) necrosis
v) wilt
vi) saprophyte
vii) obligate parasite
viii) sporangium

2. What do you call when yellowing occurs caused by lack of light?

3. What kind of pathogens normally causes mosaic?

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PRACTICAL 2

DEMONSTRATING KOCH’S POSTULATES

Isolation of suspected pathogen from the infected plant materials into pure
culture

Materials

Samples of plants/plants having leaf spot or any possible disease

Scalpel, forceps, ethanol, paper towel/tissue

PDA agar

Procedures

1. Describe and record the symptoms shown

2. Using sterile scalpel cut several small sections from the margin of the infected
leaves.
3. Place the sections from margin of lesions in 99.9% Ethanol/ 10% Clorox for few
seconds.
4. Using sterile forceps, transfer the tissue sections into sterile paper towel to
remove excess Clorox/ ethanol
5. Finally, transfer that section on nutrient media.
6. Incubate those cultures at room temperature (25-28°C) under light for 6 days to
enhance fungal growth and sporulation.

Results

Draw appropriate diagram to show main steps in experiment above.

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PRACTICAL 3

DEMONSTRATING KOCH’S POSTULATES

Isolation of suspected pathogen from culture media

Materials

Agar plate with cultured microorganisms (fungus)

Cellophane tape, scissors, glass slide, cover slip, light microscope, methylene blue.

Procedures

1. Cut a cellophane tape into small section (2-3 cm)

2. Drop a methylene blue on a glass slide.
3. Tease off a tiny piece of fungal spores using a cellophane tape.
4. Then, place that tape on a clean, sterilized glass slide.
5. Lower a clean cover slip carefully onto the drop.
6. Observe your slide under light microscope.

Results

Draw and label the fungal structures that you observed

10×

40×

100×

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Questions

1. What is disease? How can it be distinguished from other conditions?

2. What was the cause of the symptoms in your unhealthy plant? What evidence do
you have?
3. Why is it important to grow a pure culture and to inoculate with bacteria
from a single colony?
4. What if the disease-causing pathogen does not grow outside of the host?
5. How might this have changed Koch’s understanding of infectious agents?
6. Can you think of any examples?

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PRACTICAL 4

DEMONSTRATING KOCH’S POSTULATES

Inoculation of healthy plant with suspected pathogen

Materials

Seedlings of banana, culture of Rhizopus, Iron fillings, distilled water, inoculating

needle, beaker, brush

Procedures

1. Using the inoculating needle, transfer the spores from the available fungal culture
into a beaker filled with water (50 ml)
2. Mix them thoroughly so that the contents become homogenous.
3. Take one seedling and rub the leaves with Iron fillings.
4. Dip a brush into the spore solutions and brush them onto leaves, and leave the
seedling for a week.
5. Observe the leaves again the next week to see the symptoms of disease.

Results

Draw and label the procedure for this process

Questions

1. What is the importance of Koch's Postulates?

2. Why is it important in Plant Pathology?
3. Why have Koch's Postulates remained unchanged for over a century?
4. Why were you instructed to rub the banana leaves with an iron filling?
5. Other than observations of appearance, what further investigations might have
been done to prove that the organism that grew on the banana leaves in
Procedure 5 was the same one that you started with in Procedure 1?

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 RECORD WORK

Experiment : ………………………………………………… Date : …………………..

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Title : ………………………………………………….

OBSERVATIONS AND DIAGRAMS

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 RECORD WORK

Experiment : ………………………………………………… Date : …………………..

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Experiment : ………………………………………………… Date : …………………..

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Title : ………………………………………………….

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 RECORD WORK

Experiment : ………………………………………………… Date : …………………..

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Title : ………………………………………………….

OBSERVATIONS AND DIAGRAMS

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