ACTIVITY NO.

10
SOIL MICROBIOLOGY

I. Introduction:
An unpolluted environment is a habitat for a limited number of soil bacteria but a
polluted one is rich with various kind of microorganisms. In the soil habitat,
microorganisms are positive factors in the cycle of carbon, sulfur and nitrogen.
They are essential in the breakdown of organic matter to recycle the release of
these substances into the environment. Many types of bacterial activity such as
nitrogen fixation are so important to cycle back nitrogen into the atmosphere. This
activity will signify the fact as to how essential bacterial intervention is in supporting
life.
II. Objectives:
a. Appreciate the importance of soil microbes in maintaining a balance ecology
b. Identify the type of microorganism usually found in the soil
c. Enumerate factors that affect the growth of soil microbes
III. Materials:
Soil soil containers Platform balance sterile slides Urea
Erythrosin (or rose Bengal) 5% aqueous phenol Calcium chloride
microscope starch coverslip
Video Links:
https://www.youtube.com/watch?v=-dhdUoK7s2s
https://www.youtube.com/watch?v=DVxeqKyB5Tg&t=53s
https://www.youtube.com/watch?v=Wc_B0s-IW6Q
IV. Procedure:
A. BURRIED SLIDE TECHNIQUE
The “burried slide technique”, is a simple method of studying soil populations,
qualitatively more than quantitatively.
1. Collect field soil and pass it through a fine sieve.
2. Weigh out three portions of 250 grams of soil.
3. Add one gram of urea to one portion. One gram of starch to another and
nothing to the third portion, which serves as control.
4. Mix the soil and the added substances thoroughly.
5. Place each portion in separate containers.
6. Moisten the samples.
7. Insert two clean slides into the soil and pack the soil against the slides.
8. Cover the container to prevent excessive evaporation.
9. Incubate the soil at room temperature.
OBSERVE AFTER ONE WEEK.
1. Remove one slide from each container by means of a pair of forceps.
2. Wash the soil from the slide by dipping gently the slide in water.
3. Air dry the slide and fix the smear over low flame.
4. Place the slide over a steam bath and flood the stain with smear. The stain is
made up of:
Erythrosin (or rose Bengal) ………… 1 gram
5% aqueous phenol ………………… 100 ml
Calcium chloride …………………… 0.50 gram
5. Steam for 5-6 minutes.
6. Do not allow the stain to dry.
7. Wash the slide carefully to remove the soil particles and excess stain.
Dry in air and examine under oil immersion.
8. Repeat steps for the second slide a week later.
 9. Make a drawing which shows the different forms of microorganisms seen
under the microscope.
B. THE ROOT NODULES OF LEGUMES:
1. Examine and draw the root system of any available legume such as
mongo, peanut or soybean. Note the location, nature of attachment, and the
shapes and sizes of the nodules.
2. Select an effective nodule and sterilize in 10% calcium hypochlorite for 1
to 3 minutes. Rinse in sterile water. Cut and crush the nodule in sterile water
on a plain slide.
3. Transfer the loop of turbid suspension to a clean slide and allow the
smear to dry.
4. Fix and gram stain.
5. Draw the microorganisms seen from the samples
 ACTIVITY NO.10
WORKSHEET
Name/s: Baylon, Gwyneth Mae M. Date: April 24, 2022
Bunyi, Markiely A. Group no. 2
Encarnacion, Destine John V.
Galera, Samantha Nicole S.
Course and Section: BS Biology II-B

Observations:
A. Soil Populations:
Zygomycota (fungi) Protozoa Nematodes

Azotobacter (bacteria) Psedomonas (bacteria) Mycorrhizal (fungi)

 B. Observed microorganisms in the root nodules
Enisifer meliloti Rhizobium Mezorhizobium loti
leguminosarum

Bradyrhizobium elkanii Azorhizobium Burkholderia

caulinodans

Questions/Discussions:
1. What do urea and starch provide to the soil microorganism?
➢ Urea and starch work differently to give benefits to the soil microorganism.
First, urea is known as a fertilizer that give off nitrogen and this nitrogen is
needed for microbial growth. Second, starch work as a food source for these
microorganisms and is also needed for its growth.

2. What is buried slide technique?

➢ It is a technique where in the glass slides are buried in soil and incubate it
for a couple of days. This is done so that the microorganism will attach and
grow on the surface of the slides.

3. Describe the organism found in root nodules.

➢ Most of the organisms found in root nodules are nitrogen fixing bacteria.
These bacteria take in the nitrogen gas and convert it into soluble
nitrogenous compounds which is needed by the plants.

4. What is the role of these microorganisms?

➢ Nitrogen-fixing bacteria are microorganisms that can convert nitrogen from
the atmosphere into fixed nitrogen (inorganic compounds usable by plants).
These species are responsible for more than 90% of all nitrogen fixation,
and hence play a vital part in the nitrogen cycle. The purpose of nitrogen-
fixing bacteria is to provide plants with a food that they can't get from the
 air. Microorganisms that fix nitrogen do what crops can't: they get
assimilative nitrogen for them. Bacteria absorb it as a gas from the air and
release it to the soil as ammonia. Plants can only ingest nitrogen from the
soil in the form of nitrogenous inorganic compounds, which explains the
need of nitrogen fixation.

5. Explain the importance of having healthy soil.

➢ Healthy soils yield healthy crops, which nourish both humans and animals.
Food quality and quantity are inextricably tied to soil quality. Soils provide
our food-producing plants with the critical nutrients, water, oxygen, and root
support they require to grow and thrive. Healthy soils have a stronger ability
to absorb and keep water, reducing evaporation and increasing drought and
extreme weather resilience. The nutrient density of meals is increased when
soil bacteria provide nourishment for plants instead of synthetic fertilizers,
which benefits public health. Crops are better equipped to combat pests and
disease, decreasing the need for costly chemicals - this is beneficial for
farmers' bottom lines while also helping to reduce pollution, resulting in
healthier watersheds and water quality. Healthy soils also promote
biodiversity, provide better habitat for wildlife and pollinators, and absorb
carbon from the atmosphere.

6. What would happen to plants if there were no microorganisms in soil?

➢ Plants, as we all know, rely on microorganisms to live. They will swiftly lose
their ability to produce energy via photosynthesis if they do not have
microorganisms to take in and transform essential chemical components
into useful bits.

Conclusion:
Microbes in the soil play a key role in nutrient recycling, particularly carbon,
nitrogen, phosphorus, and sulfur. Bacteria are a type of microbe that helps to maintain
soils healthy and productive. Urea is known as a fertilizer that give off nitrogen and this
nitrogen is needed for microbial growth and starch work as a food source for these
microorganisms and is also needed for its growth. Buried slide technique is a technique
where in the glass slides are buried in soil and incubate it for a couple of days. This is
done so that the microorganism will attach and grow on the surface of the slides. Most of
the organisms found in root nodules are nitrogen fixing bacteria. These bacteria take in
the nitrogen gas and convert it into soluble nitrogenous compounds which is needed by
the plants. Nitrogen-fixing bacteria are microorganisms that can convert nitrogen from the
atmosphere into fixed nitrogen (inorganic compounds usable by plants). Healthy soils
yield healthy crops, which nourish both humans and animals. Healthy soils have a
stronger ability to absorb and keep water, reducing evaporation and increasing drought
and extreme weather resilience. It also promotes biodiversity, provide better habitat for
wildlife and pollinators, and absorb carbon from the atmosphere. Soil depth, organic
content, porosity, oxygen and carbon dioxide concentrations, soil PH, total phosphorus
and other factors all influence the microbial population.