amna mushroom report
TRANSCRIPT
INTRODUCTION:
National Agriculture Research Centre (NARC) Islamabad:
National Agricultural Research Centre (NARC), Islamabad established in 1984, is the largest
research centre of the Pakistan Agricultural Research Council (PARC). NARC, with a total land
area of approximately 1600 acres, is located near Rawal Lake, six kilometers south-east of
Islamabad. Physical facilities in term of experimental fields, laboratories, green houses, gene
bank, library/ documentation, auditorium, machinery & lab equipment repair workshops, stores,
hostels, cafeteria, audio visual studios, are also available at NARC.
NARC coordinated programs serve as a common platform for the scientists working in
different federal, provincial agricultural research, and academic institutions to jointly plan their
research activities, avoiding unnecessary duplication of research efforts. Research which can best
be addressed at a National Centre rather than by provincial institutions is undertaken at NARC.
The adaptation of technologies available from the international research system is also managed
by NARC, in collaboration with the provincial research and extension institutions. In particular,
research requiring sophisticated instruments like electron microscopes, ultracentrifuges, and
elaborate analytical and quality testing facilities is undertaken at NARC, supported by highly
qualified and trained manpower.
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Crop Diseases Research Institute:
Crop Diseases Research Institute (CDRI) is the premier Institute in Pakistan working on Crop
Health Management through applied and basic research in phytopathology under the aegis of
Pakistan Agricultural Research Council. It existed in its nuclear form since 1955 and through
several reorganizations, attained the structure and mandate of Crop Diseases Research Institute
in 1983. Later in 2001 in an institutional re-organization of PARC it merged into a newly
developed Institute of Plant and Environmental Protection with the change of status from
Institute to Program. The Institute is again elevated from program to Institute in 2013. It is the
only one of its kind in the country. It is working with the infrastructure of main laboratories at
National Agricultural Research Centre (NARC), Islamabad and a substation at Murree.
Introduction to Oyster Mushroom:
Mushroom in general and Pleurotus in particular are an important source of nutrition particularly
for the people on cereal-based diet. Edible species of mushroom are low in calories, fats, sodium,
carbohydrates and cholesterol, whereas, rich in proteins, minerals, vitamins and fibers (Nasim et
al. 2001). Besides nutritional importance genus Pleurotus is also known for its medicinal value
(Kothe 2001), paper pulp bleaching, cosmetics and industrial use (Kyung-Ho Ma et al. 2009;
Sigoillot et al. 2005).
The mushrooms are naturally grown in fields, forests; on manure heaps, water channels and hilly
areas, mostly during and just after rains. The most popular varieties are Agaricus bisporus
(European or white button mushroom), Pleurotus spp., (Oyster mushrooms or Dhingri),
Volvariella volvacea (Chinese or paddy straw mushroom) Lentinu sedodes (Shiitake mushrooms)
and Auricu slaria (Black ear mushroom).
Mushrooms are valuable food, which are low in calories, high in vegetable proteins, zinc, chitin,
fiber, vitamins and minerals (Alam and Saboohi, 2001). The mineral salt content in mushrooms
is superior to that of meat and fish and nearly twice that of the most commonly used vegetables
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(Chang, 1972). Anticancer medicine (Leutinan) is produced recently by some chemical
companies from the extract (Polysaccharides) of Shitake mushroom (Mori, 1986).
The Oyster mushroom is grown under natural conditions on living trees as parasite or dead
woody branches of trees as saprophyte and primary decomposer. The chemical composition of
the fresh fruiting bodies of oyster mushroom, Pleurotus ostreatus indicates a large quantity of
moisture (90.8%), whereas fresh as well as dry oyster mushrooms are rich in proteins (30.4%),
fat (2.2%), carbohydrates (57.6%), fiber (8.7%) and ash (9.8%) with 345 K (cal) energy value on
100 g dry weight basis; while vitamins such as thiamin (4.8 mg), riboflavin (4.7 mg) and niacin
(108.7 mg), minerals like calcium (98 mg), phosphorus (476 mg), ferrous (8.5 mg) and sodium
(61 mg) on 100 g dry weight basis, are also found present (Pandey&Ghosh, 1996). Rambelli &
Menini (1985) reported that this mushroom is reputed to be antitumoral because of its chemical
composition.
The oyster mushrooms can be cultivated successfully under semi controlled conditions in a small
space by using agricultural as well as industrial waste and other refuse as substrate. Badshahet
al., (1992) have grown Pleurotus ostreatus on wheat straw, sugarcane bagasse, corn cobs or
sawdust by mixing 120-130 g of spawn with 2 kg of substrate and placing the mixture in
sterilized polyethylene bags which were kept in the dark at 25 ͦ C for 2-3 weeks.
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Varietals difference was observed for mycelial colony proliferation of mushroom (Khandakar,
2004). Some growth regulators at different concentrations affect the yield and size of mushroom
(Charles, 1986). Plant growth regulators and media can also play an important role for in vitro
mycelial colony proliferation of mushroom (Muniruzzaman, 2004). Quite a large number of
experiments have so far been carried out throughout the world to find out the appropriate variety
and media for mushroom cultivation. However, in the country sufficient research work have not
yet been done on different variety and media for mushroom cultivation. The present piece of
research work was, therefore, undertaken to investigate the mycelial colony proliferation of
Oyster mushrooms on PDA (Potato Dextrose Agar), CMA (Corn Meal Agar), TWA (Tap Water
Agar) and MEA (Malt Extract Agar) media.
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MATERIAL AND METHODS:
Sources of materials:
The species of oyster mushroom (strain AG 572) was obtained from Crop Disease Research
Institute (CDRI), Department of Plant and Environmental Protection (DPEP), NARC, Islamabad
and maintained in potato dextrose agar (PDA), corn meal agar (CMA), tap water agar (TWA),
and Malt Extract Agar (MEA) media.
Affect of different Media and Temperature on the growth of mycelium:
The effects of culture media and temperature on the mycelium growth of oyster mushroom
Pleurotus ostreatus for spawn production were studied.
Several trials were designed to evaluate factors affecting mycelium growth of oyster mushroom.
For each test, mycelium discs with similar diameter (1 cm) were used. The discs were obtained
by using the sterilized punch-hole tool to cut the mycelium and place into a new sterilized
medium plate by using a transplant needle. The diameter of the mycelium extension was
measured every 2 days with the help of a transparent ruler at the same time (between 9:00 AM
and 12:00 PM) and surface mycelium density was observed.
Effect of culture media:
Four agar media were used including (PDA) potato dextrose agar medium, (CMA) corn meal
agar medium, (TWA) tap water agar medium (MEA) malt extract agar medium. The ingredients
for those culture media were as follows:
PDA:
200 g potato, 20 g dextrose, 15 g agar powder, and 1000 ml distilled water;
CMA:
30 g corn meal, 20 g dextrose, 15 g agar powder, and 1000 ml distilled water;
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TWA:
15 g agar powder and 1000 ml distilled water;
MEA:
20 g malt extract, 20 g dextrose, 15 g agar powder and 1000 ml distilled water.
Media and Petri dishes (10 cm diameter) were autoclaved at 121℃ (at a pressure of 1.3 kg/cm2)
for 20 min. The mycelium discs (1 cm diameter) of oyster mushroom was placed in Petri dishes
containing each culture medium (20 ml) under aseptic condition and incubated at ±25ºC &
±28℃ temperature in the darkness. The diameter of the mycelium expansion was measured
every 2 days for 8 days.
Effect of temperature:
The Petri dishes containing sterilized (PDA) potato dextrose agar medium, (CMA) corn meal
agar medium, (TWA) tap water agar medium (MEA) malt extract agar medium (20 ml) each
were inoculated with the mycelium discs (1 cm diameter) of oyster mushroom under aseptic
condition and incubated in the darkness at two levels of temperature (25℃ and 28℃). The
diameter of the mycelium expansion was measured every 2 days for 8 days.
Experimental design:
The experiment was conducted in Crop Disease Research Institute (CDRI), Department of Plant
and Environmental Protection (DPEP), NARC, Islamabad during summer 2015 (June to July).
The experiment was arranged in a completely randomized design (CRD) with three replications
per treatment.
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RESULT AND DISCUSSION:
Effect of different culture media on the mycelium growth of oyster mushroom:
Every living being requires food for its growth and reproduction and oyster mushrooms are not
an exception to it. In order to culture the oyster mushroom in the laboratory, it is necessary to
furnish those compounds in the media which are requires for its growth and other life process. In
the present study, oyster mushrooms PO was cultured successfully in the laboratory
environment. During the present investigation, different media, i.e., PDA, CMA, TWA, and
MEA were used to identify their effect on mycelium growth.
The observed mycelium colony diameter of mushroom PO on four different media significantly
differed with each other. The Mycelium density of oyster mushroom PO was compact in all
media. The average mycelium diameter of oyster mushroom ranged from 3.92 ± 0.05 to 9.00 ±
0.00 cm at 8 days after inoculation. The mycelium growth of mushroom PO on PDA and CMA
media was better than those on TWA and MEA media at 2, 4, and 6 days after inoculation
(DAI); however, they were not significantly different at 8 DAI.
The result indicated that these four culture media are suitable for mycelium growth of mushroom
while PDA and CMA media are more suitable for the mycelium growth of mushroom PO. This
may be due to availability of required nutrients for mushroom PO in PDA and CMA media.
OYSTER MUSHROOM MYCELIUM ON PDA OYSTER MUSHROOM MYCELIUM ON CMA
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OYSTER MUSHROOM MYCELIUM ON MEA OYSTER MUSHROOM MYCELIUM ON TWA
The results of the present research showed that the mycelium of PO took shorter time to
complete growth in all media. These findings were similar to the results reported by Mshandete
and Mgonja; Mansur et al. They indicated that PDA and CMA were suitable media for culture
of Pleurotus species to achieve high level mycelium biomass, exo-polysacharides and mycelium
protein. In Pakistan, potato is so much cheaper than corn malt extract, so PDA medium was more
suitable and efficient to use as alternatives for culture of mushroom PO. From this result, we
chose local materials as cheaper materials to prepare media for culture of mushroom PO.
Effect of temperature on mycelium growth of oyster mushroom:
Temperature is a very important environment factor for mycelium growth of fungi. To determine
optimal temperature for mycelium growth, of oyster mushroom was cultivated in PDA, CMA,
TWA, and MEA medium at two temperatures (25℃and 28℃). The trend of mycelium growth
in response to temperature was not very similar, where the optimum temperature for oyster
mushroom was found to be 25℃ followed by 28℃.The mycelium growth of mushroom PO was
significantly faster at 25℃. The temperature tested to grow after 8 days inoculation at 25℃ and 28℃. The mycelium density of mushroom PO was very thin at 28℃ indicated that the
optimum temperature for mycelium growth of oyster mushroom was 25~30℃. This optimum
temperature result indicated that the oyster mushroom PO was able to grow better in summer and
autumn season in subtropical and tropical regions as potential opportunity to develop oyster
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mushroom production in poor and developing countries in Asia. The result is quite similar to the
results reported by Kashangura, (2008) and Choi et al.(2003). When demonstrating that the
mycelium growth and fruiting formation of oyster mushroom species were affected by
temperature and they could grow at high temperature as summer season in tropical regions.
GROWTH OF MYCELIUM INCUBATED AT 25℃ ON PDA. GROWTH OF MYCELIUM INCUBATED AT 28℃ON PDA.
GROWTH OF MYCELIUM INCUBATED AT 25℃ON CMA. GROWTH OF MYCELIUM INCUBATED AT 28℃ON CMA
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GROWTH OF MYCELIUM INCUBATED AT 25℃ON MEA. GROWTH OF MYCELIUM INCUBATED AT 28℃ON MEA
GROWTH OF MYCELIUM INCUBATED AT 25℃ON TWA. GROWTH OF MYCELIUM INCUBATED AT 28℃ON TWA.
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CONCLUSION:
In conclusion, the mycelium growth of oyster mushroom PO was affected by different culture
media and temperature.
It was concluded that PDA, CMA, TWA, and MEA were the most favorable media for the
mycelium growth of mushroom while PDA and CMA were the most suitable media for
mycelium growth of oyster mushroom PO.
Maximum mycelium growth of oyster mushroom PO was achieved by cultivating them at the
optimum temperature of 25℃.
In all studied cases, mycelium growth of oyster mushroom PO was significantly better on PDA
and CMA at 25℃.
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