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MEDIA AND CULTURE

30.10.2015

Group B
Jessica De Marco Matr. 21544096
Shanta Subba 21228261
Table of contents

1. INTRODUCTION

Like other organisms fungi also need nutrient to grow and survive in the nature. The artificial
media contain all essential elements required for fungi to growth in the laboratory. These
elements are macro and micro elements. From the name itself one can understand that macro
elements for instance carbon, oxygen, nitrogen, hydrogen, sulfur, phosphorus, potassium, sodium
and magnesium are taken by fungi in a large amount whereas micro elements like zinc, copper,
calcium, manganese or iron in a small quantity. The quantity and the variety of elements up taken
by fungi vary from species to species and are directly proportional to the fungal growth.
Trametes versicolor is widespread and common white rot fungi. It is known to produce laccase
that has property to degrade lignin. The fruiting bodies develop as overlapping clusters on dead
branches, stumps and logs (Stephenson, 2010).
In this study we compared growth of Trametes versicolor on different media and analyzed the
optimal media for its growth.
Also in this experiment we isolated small parts from inner (lamina) and outer of fruiting body of
Agaricus bisporus and Pleurotus ostreatus and inoculated them on the agar plate to see their
growth and the extent of contamination that could occur by inoculating fungal pieces from the
wild.

2. MATERIAL AND METHODS

The mycelia of Trametes versicolor grown on the agar plate and liquid media are used as an
inoculum to inoculate in different media as follows.
1. Malt-Agar
2. Millet-cultures
3. Straw-cultures
4. Liquid BSM Media

For the isolation of sample pieces, fruiting bodies of Agaricus bisporus and Pleurotus ostreatus
were taken.
The preparation of media and inoculation of fungus were performed according to the protocol (A.
Majcherczyc, O. Voigt and M. Zomorrodi, 2015). After 7 days of incubation at 37o in the dark, the
inoculated mycelia of all three given fungus were monitored.
3. RESULTS

3.1. Small block of Trametes versicolor grown on agar was taken to inoculate on agar plate.
The observation made after 7 days of incubation showed fully grown mycelia on the agar
surface. However, the mycelia grown looked thin and less dense.

FOTO Fig 1:

3.2. Three blocks of Trametes versicolor agar were taken to inoculate to the liquid medium.
After 7 days of incubation at 37o in the dark, mycelia were seen to be grown like patches
circling each inoculum.

Fig 2: The liquid culture from agar inoculum.


3.3. Trametes versicolor grown on liquid media was used for inoculation into the liquid
medium and incubated at 37o in the dark. The mycelial growth after 7 days showed
homogenous growth on the surface of medium without forming any patches.

Fig 3: Liquid culture from liquid inoculum.

3.4. Three blocks of Trametes versicolor agar were inoculated to the millets half-filled in the
flask and incubated like the previous cultures. At day 7 mycelia were seen to grow fast and
dense giving a cottony look engulfing millets.

Fig 4: Millets culture from agar inoculum.


3.5. Here Trametes versicolor already grown on millets were used as an inoculum (one table
spoon full) to grow them on straw. Observation made on day 7 of incubation showed
production of large quantity of biomass.

Fig 5: Straw culture from millets inoculum.

3.6. Three blocks of Trametes versicolor agar were inoculated to straw. The result after 7 days
showed very slow growth of mycelia on the straw.

Fig 6: Straw culture from agar inoculum.

3.7. Isolated pieces of fruiting bodies were observed and Pleurotus ostreatus inoculated on
agar plate showed good growth whereas Agaricus bisporus showed poor growth with
contamination.
From the above observations we can conclude that mycelial agar blocks inoculated showed better
growth on the agar plate and not on the liquid media and on straw. However, the homogenous
mixture of liquid mycelial media looked uniformly grown on the liquid media. And mycelia grown
on millets seem to grow well and produce abundant biomass on straw.

4. DISCUSSION

Fungi can be grown on different media under different condition depending upon their
adaptability to the ingredients present in the medium. Therefore, the media have been
categorized into synthetic, semi-synthetic and complex media according to the nutrients present
in them.
In our experiments, we used malt-agar, millet and straw as a complex media and BSM liquid as a
semi-synthetic medium. The sources of carbon and nitrogen in agar media are the sugar present in
the malt extract and the proteins contained in grains, respectively. Similarly, in millets protein
produced is again a source of nitrogen and starch as a carbon source. In case of BSM, glucose is
added as a carbon source and L-asparagine as a nitrogen source. In straw, cellulose and
hemicellulose are the main components that can be used as a carbon source. Structural proteins
of the primary cell-wall of the live plant remain as a part of lignified cell-walls in straws. These
proteins can be used as nitrogen source.
The mycelial growth on different media using different inoculum styles showed comparable
growth in all media. Agar block inoculated to agar plate and to millets showed good mycelial
growth but not to liquid media and straw. The reason behind this could be the presence of fewer
amounts of mycelia in the inoculum block which would not be sufficient for hyphae to grow and
form a network. However, inoculation of liquid mycelia to liquid medium showed uniform growth
over the liquid media and a spoon full of mycelial millets inoculated to straw showed good growth
and production of large amount of biomass.
From these observations, we can conclude that fungal growth is highly dependent on different
ways of inoculation and the substrate used.
During the isolation of fruit body it was observed that the sample used from the inner part
(lamina) grew better on the agar surface than from the other parts of mushroom. However,
Pleurotus ostreatus showed better mycelial growth than Agaricus bisporus whose plates got
contaminated after few days of mycelial growth.

5. LITERATURE

Stephenson, S.L., 2010, The Kingdom Fungi: The Biology of Mushrooms, Molds, and
Lichens, Timber Press