On Friday, Dr Wan asked us to do flowchart about how to isolate fungi in groups. My group did on isolation of fungi from tropical soil. The purpose of isolate of fungi from tropical soil is to isolate secondary metabolites such as antibiotic, enzyme, protein and so on.
Fungi produced two types of metabolites
Primary metabolites
Fungi produced primary metabolites for their proper growth, reproduction and their proper development.
Secondary metabolites
While secondary metabolites fungi produce for their own defense, to grow in the same medium and to killed other microbes in the same medium.
Name of the culture medium used for fungi growth
- Brain heart infusion agar
- Czapek yeast agar
- Czapek yeast broth
- Inhibitory mold agar
- Mycosel/Mycobiotic agar
- Potato Dextrose agar
- Sabouraud's heart infusion agar
- Bannerot synthetic media agar
- Potato flake agar
- Water agar
ISOLATION OF FUNGI FROM TROPICAL SOIL
- Prepare Potato Dextrose Agar (PDA)
- Dig earth surface at 10-12 cm depth and collect soil.
- Perform serial dilutions.
- Inoculate sample using spread plate method on the PDA.
- Incubate the agar plate in inverted positions at 28⁰C for 48 hours.
- Subculture the single colony into new PDA to isolate pure culture.
Isolation of fungi in bread mold
https://www.ijcmas.com/vol-4-1/Unachukwu%20M.N%20and%20Nwakanma%20C..pdf
Isolation of fungi from seawater
www.mdpi.com/2309-608X/3/3/36/pdf
FLOW CHART OF ISOLATION GUAVA’S TREE DISEASE
1.
Samples were collected from infected tissues to
ascertain the association of different pathogens. The samples consisting of
bark at collar region, twigs and soil were collected. Trees showing declining
symptoms were selected and samples of roots, soil, bark at collar region, and
leaves of healthy as well as diseased plants were collected.
2.
After that, blench all the disease’ parts by
using mortar into a powder. The infected samples (in form of powder) are mix
with distilled water.
3.
Infected samples are plated on PDA by using the
serial dilution technique. The plate that need to be inoculated are in a several
dilution factor by using spread plate (NOTE: try to inoculate the sample in
higher diluting factor).
4.
Incubated all the plates in incubator at 27 °C
for 3‒7 days and data was recorded. The fungi which colonized on these pieces
was purified and identified on the basis of their morphological characters.
5.
Cut the small pieces of fungus by using scalpel,
that differ by the morphology into a new plate. Place it on the middle plate
and incubated the plate again in suitable temperature again (27 °C)
for another week. For the type of fungus that bring the disease, Botryodiplodia theobromae, the mycelial
growth of isolates was classified as fluffy or depressed, uniform to irregular
and cottony white turning to black.
6.
After a week, observe the morphology of fungus
and record in your Results 
