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IN VITRO CONTROL OF ASSOCIATED MYCOFLORA OF OYSTER MUSHROOM

SUBSTRATES BY THE APPLICATION OF FUNGICIDES

Article · January 2009

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 BANGLADESH RESEARCH PUBLICATIONS JOURNAL
ISSN: 1998-2003, Volume: 2, Issue: 4, Page: 737-741, July - August, 2009

IN VITRO CONTROL OF ASSOCIATED MYCOFLORA OF OYSTER

MUSHROOM SUBSTRATES BY THE APPLICATION OF FUNGICIDES

Pervez, Z.∗1, Bhuiyan, M.K.A.2 and Islam, M.S.1

Pervez, Z., Bhuiyan, M.K.A. and Islam, M.S. (2009). In Vitro Control of Associated Mycoflora of Oyster
Mushroom Substrates by the Application of Fungicides. Bangladesh Res. Pub. J. 2(4): 737-741.
Retrieve from http://www.bdresearchpublications.com/admin/journal/upload/08081/08081.pdf

Abstract
A laboratory trial has been conducted with Formalin, Bavistin and
combination of Formalin and Bavistin at three different concentrations
against identified associated mycoflora of Oyster mushroom substrates. The
combination of Formalin and Bavistin at the highest concentration (500+75
ppm) was found to be the best in inhibiting the radial colony growth of all
the identified fungi.
Key words: mycoflora, oyster mushroom, fungicide.
Introduction
Mushrooms are large reproductive structure of edible fungi belonging to
Basidiomycotina. They are non-green fungal plants occurring seasonally all over the world
in various habitats varying from sandy plains to thick forests or green meadows to roadside
pathways. They comprise a large heterogeneous group having various shapes, sizes,
colours; appearance and edibility (Chang and Miles, 1991). There are 10,000 different
types of mushrooms of which 2000 are edible. Of the various kinds, oyster mushroom
scientifically known as Pleurotus ostreatus is most suited to the climatic condition of
Bangladesh.
The economic importance of the mushroom lies primarily in its use as food for
human consumption It is a good source of protein, vitamins and minerals (Khan et al.,
1981) and contain about 85-95% water, 3% protein, 4% carbohydrates, 0.1% fats and 1%
minerals and vitamins (Tewari, 1986). Mushroom protein is intermediate between that of
animal and vegetable (Kurtzman, 1976) and the amount of niacin, pantothenic acid and
biotin are of appreciable level (Subramanian, 1986).
Mushrooms have medicinal properties like anti-cancerous, anti-cholesteral and
anti-tumorous activities and are useful against diabetes, ulcer and lungs diseases. The folic
acid present in oyster mushrooms helps to cure anemia. It is suitable for people with hyper-
tension, obesity and diabetes due to its low sodium: potassium ratio, starch, fat and
calorific value. Alkaline ash and high fibre content makes them suitable for consumption
for those having hyperacidity and constipation. A polycyclic aromatic compound
pleurotin has been isolated from P. griseus which possess antibiotic properties (Quimio,
1976).
The management of mycoflora associated with substrate of mushroom is very
difficult because both the host and the parasites are fungi. Chemical treatments with 1%
formaldehyde and that using 2.0% copper sulphate gave satisfactory mushroom yields.
Sterilization method had little effect on the rate of mycelial development. Chemical

∗ Corresponding Author’s Address: Email: zehadpervez@yahoo.com

1Department of Plant Pathology, Patuakhali Science and Technology University, Dumki, Patukhali.
2Department of Plant Pathology, BSMRAU, Gazipur1706, Bangladesh.
Pervez et al. 738

disinfection is recommended as it is cheaper than steam sterilization (Afyon, 1988).

Different fungicide treatments namely Benlate (50% benomyl), Fermenter Forte (35%
carbendazim) or Sportak Alpha (30% prochloraz + 8% carbendazim) at 100-250 ppm,
favourably influenced mycelial growth and in some cases increased fruit body yield
(Lelley et al. 1991). Maximum control of green mould in Oyster mushroom was obtained
when thiabendazole at 2 or 5 g/m2 was applied to the substrate before pasteurization.
The highest yield of oyster mushrooms was obtained with the 5 g/m2 treatment (Jhune et
al. 1990).
Rai and Vijay (1992) demonstrated that carbendazim stimulated the mycelial
growth of P. sajor-caju at low conc. (5 and 10 ppm) but inhibited it at 25 ppm with
complete inhibition at 100 ppm. T. viride completely inhibited the growth of P. sajor-caju
when the fungi were inoculated together, but had no effect if inoculated after P. sojor-
caju had been growing for 7 or more days. In mixed cultures, 5 ppm carbendazim
completely inhibited T. viride and stimulated P. sajor-caju. The yield of P. sajor-caju was
highest with Bavistin-treated straw (Bermudez 1994). Highest yield (95% biological
efficiency) was recorded with substrates treated with Bavistin [carbendazim] at 75 ppm +
formalin [formaldehyde] at 500 ppm, which was equivalent to the yields obtained from
substrates pasteurized with hot water (70-80 C) or Bavistin at 75 ppm (Pani et al. 1998).
Mushroom production is greatly affected by pests and diseases. The most
notorious of which is Trichoderma that is resistant to pasteurization. Results of the basic
tests showed that benomyl 1000, 500 and 250 ppm; prochloraz 1000 ppm; and imazalil
1000 ppm, 500 ppm inhibited the growth of Trichoderma. Moreover, chemical treatment
with acids and bases showed that acetic acid, salicylic acid, potassium hydroxide and
sodium hydroxide were effective at 2000 ppm. Some plant extracts and antibiotics have
the potential too of controlling Trichoderma contamination in mushroom cultivation. This
was also proven by the confirmation tests where 50 and 100 ppm concentrations of
fungicides inhibited the growth of Trichoderma (Domondon and Poppe 2000). Among the
tested three disinfectants, Environ and Purogene were more effective than Sudol in
limiting the recovery of Trichoderma spp. and Cladobotryum dendroides. All three
disinfectants applied 12 h after artificial contamination onto wood, concrete or glazed
surfaces were able to reduce the recovery of these isolates more effectively than when
they were applied 12 h before artificial contamination. In many cases, the protection
achieved by the physical nature of the smooth glazed tile surfaces was equivalent to that
available from the application of disinfectants, thus highlighting the attention needed to
the nature of building surfaces in the structures used in the mushroom industry (Abosriwil et
al. 2002).
To overcome the discrepancy among the effect of fungicides and their
concentrations, suitable fungicides at optimum concentration is needed to control
different mycoflora associated with oyster mushroom to increase the growth and yield. In
Bangladesh, no published reports are available in controlling the mycoflora associated
with the mushroom.
Keeping these in view, the present studies were undertaken on the basis of the
following objectives:
i. To find out the appropriate chemicals and their doses for controlling
associated mycoflora with Oyster mushroom sbstrate.
Materials and Methods
A total of 50 spawn packets with colonized substrate of P. ostreatus were collected
randomly at different growth stages from Mushroom Culture Center, Savar, Dhaka. Ten
mycoflora including Aspergillus flavus, A. fumigatus, A. niger, A. nidulans, A. terreus,
Penicillium citrinum, P. thiersii, Penicillium. sp., Rhizopus stolonifer and Trichoderma
harzianum were isolated and identified as associated with P. ostreatus (Oyster mushroom)
substrate.
An attempt was made to control different mycoflora associated with substrate
colonized Oyster mushroom with different fungicides in vitro. The experiments were
conducted at National Mushroom Development & Extension Centre, Sobhanbagh, Savar,
Dhaka, Bangladesh during 2006-2007.

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In Vitro Control of Associated Mycoflora of Oyster Mushroom 739
Laboratory evaluation of fungicides against associated mycoflora from different substrate
of P. ostreatus
Three different concentrations namely 100, 250 and 500 ppm of Formalin, 25, 50
and 75 ppm of Bavistin and 100 +25, 250 + 50 and 500 + 75 ppm of Formalin + Bavistin
were tested in vitro to evaluate their effect on the radial growth of isolated fungi. Details
of the fungicides are given in the Table 1.
Table 1. Details of the fungicides
Trade IUPAC Name Chemical name Molecular Active ingredient (%)
name Formula
Formalin Methanol Formaldehyde C-H2O 50
Bavistin 50 Methyl(N-(1 H- Carbendazim C9H9N3O2 50
WP benzoimidazol-
2-yl)carbamate
Formalin Methanal Formaldehyde C-H2O 100% Formalin
+ + + + contains40%
Bavistin 50 Methyl(N-(1 H- Carbendazim C9H9N3O2 formaldehyde and
WP benzoimidazol- Bavistin contain 50%
2-yl) carbamate carbendazimas
wettable powder
Colony of the isolated fungi from selected 50 packets of wheat grain colonized
substrate of P. ostreatus were cultured on PDA. Each of the isolated fungus was treated
against the selected three chemicals at three different concentrations separately
following poison food technique (Begum 2000). The required amounts of fungicides were
mixed with melted PDA. Control was taken into the design of experimentation by keeping
untreated colony on PDA medium. Efficiency of chemicals at different concentrations
were evaluated on the basis of percent inhibition of the radial colony growth of the
isolated fungi, and computed as a proportional measurement of colony diameter on
control plate expressing in percentage, following the formula given by Sundar et al., 1998.
X−Y
% Inhibition = X100
X
Where, X= radial growth (mm) of control plates,
Y = radial growth (mm) of fungicide treated plates.
Results and Discussion
Laboratory evaluation of fungicides against associated mycoflora from different substrates
of P. ostreatus.
The results of the laboratory evaluation of Formalin, Bavistin and Formalin + Bavistin
at three different concentrations are presented in the Table 2. The highest concentrations
of Formalin, Bavistin and Formalin + Bavistin are appeared to be superior in inhibiting the
radial growth of all the tested 10 mycoflora ranging from 85.80 to 100% inhibition. Among
the tested fungicides, the combination of Formalin and Bavistin was found to be superior
at all the selected concentrations in comparison to the individual fungicide Formalin and
Bavistin. At the lowest dose of combined Formalin and Bavistin the highest 80.00%
inhibition was found against R. stolonifer followed by T. harzianum (73.86%) while the
minimum 44.11% inhibition was observed against Penicillium sp. In case of all the isolated
fungi above 92.00% inhibition was obtained at the maximum concentration of Formalin
and Bavistin combination. The complete inhibition of Aspergillus niger was observed at the
highest concentration when Formalin and Bavistin was used together.
At the lowest dose of Formalin, the maximum 78.00% inhibition was achieved
against Rhizopus stolonifer followed by Trichoderma harzianum (69.28%), while the
minimum 39.20% inhibition was recorded against Penicillium sp. The highest 96.80 %
inhibition of T. harzianum followed by A. niger (94.70%), R. stolonifer (94.00%) was
achieved at the maximum concentration of Formalin while the minimum 85.80% inhibition
was obtained against Penicillium sp.
In general, Bavistin was found to be superior in comparison to Formalin but inferior
to the combination of Formalin and Bavistin at all the selected concentrations. At the

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Pervez et al. 740

lowest dose of Bavistin, the highest 84.00% inhibition of the radial growth of R. stolonifer
was observed followed by T. harzianum(70.66%) while the lowest 44.65% reduction was
obtained against P. thiersii. The maximum 97.52% inhibition of A. nidulans followed by R.
stolonifer (97.00%) was obtained at the highest concentration of Bavistin while the
minimum 91.25% inhibition was observed against P. thiersii.
However, the results of the present study prevailed that both the fungicides
Formalin and Bavistin are effective against the associated molds of Oyster mushroom but
when these fungicides are used in combination were highly effective which are in
agreement with several investigators ( Rai and Vijoy 1992, Bermudez 1994, Anandh et al.
1998 and Pani et al. 1998).
Table 2. Laboratory evaluation of chemicals against mycoflora associated with
P. ostreatus grown substrate

Fungici % inhibition
des A. A. A. A. A. P. P. P. R. T.
With flavus fumigatus niger nidulans terreus citrinum thiersii sp. stolonifer harzianum
Conc.
(ppm)
Formalin
100 46.15 44.45 61.19 59.23 57.33 40.21 45.52 39.20 78.00 69.28
250 64.94 66.75 83.54 82.52 80.25 72.24 70.96 73.52 86.00 84.87
500 89.74 87.35 94.70 93.16 92.21 91.24 86.33 85.80 94.00 96.80
Bavistin
25 51.89 49.54 64.72 63.14 62.46 45.64 44.65 40.08 80.00 70.66
50 64.10 63.20 82.66 81.26 79.56 69.45 69.58 71.08 85.00 78.00
75 92.74 91.50 97.00 97.52 96.89 91.52 91.25 92.14 97.00 97.25
Formalin
+Bavistin
100+25 56.40 55.62 67.89 66.23 67.33 46.24 47.15 44.11 84.00 73.86
250+50 77.35 76.23 80.29 78.21 76.58 76.33 75.32 77.94 95.00 90.13
500+75 94.02 93.25 100.0 98.42 98.26 92.45 93.4 93.82 98.00 98.17
0
Untreat
ed 3.90 3.7 5.67 4.10 3.80 3.3 3.4 3.4 8.50 7.27
control (cm) (cm) (cm) (cm) (cm) (cm) (cm) (cm) (cm) (cm)

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