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Fungal interaction between Trichoderma spp. and Pleurotus ostreatus on the

enriched solid media with licorice Glycyrrhiza glabra root extract

Article  in  Acta Ecologica Sinica · March 2018

DOI: 10.1016/j.chnaes.2017.08.001

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journal homepage: www.elsevier.com

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Fungal interaction between Trichoderma spp. and Pleurotus ostreatus on the
enriched solid media with licorice Glycyrrhiza glabra root extract
Behar Moqdad Al-Ania, Mustafa Nadhim Owaidb, c, ⁎, Sajid Salahuddin Salem Al-Saeedia

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a
Department of Biology, College of Science, University of Anbar, Ramadi, Anbar 31001, Iraq
b
Department of Heet Education, General Directorate of Education in Anbar, Ministry of Education, Hit, Anbar 31007, Iraq
c
Department of Ecology, College of Applied Sciences, University of Anbar, Hit, Anbar 31007, Iraq

ARTICLE INFO ABSTRACT

Article history: The aim of this study is to investigate the antifungal activity of mycelia of Pleurotus ostreatus (white oys-
Received 22 June 2017

D
ter mushroom) and licorice (Glycyrrhiza glabra) root extract against three undesirable fungi. They are Tri-
Received in revised form 24 July 2017 choderma spp., Trichoderma harzianum I and Trichoderma harzianum II which was tested on PSA (potato
Accepted 7 August 2017
sucrose agar) medium enriched with licorice (Glycyrrhiza glabra) root extract (PSA-G media) using three
Available online xxx
concentrations (0.05, 0.10 and 0.20 g/L) in alone and dual cultures. Trichoderma spp. showed less mycelial

Keywords:
Oyster mushroom
Pathogenic fungi
Antagonistic activity
TE
growth of 8.75, 9.17 and 9.50 mm/day on PSA-G0.05, PSA-G0.1 and PSA-G0.2 respectively compared with
10.25 mm/day on fresh PSA (control) in dual culture. The best mycelial growth inhibition was recorded on
PSA-G0.2 (14.97%) by T. harzianum II in alone culture opposite 63.72% in dual ones. The lower mycelial
growth rate of T. harzianum I was 17.75 mm/day on PSA-G0.1 (0.10 g/L). In dual culture, overgrowth time
of T. harzianum I had 5 days compared as approx. 6 days in alone culture. Generally, when the concentration
Liquorice extract of licorice extract increased, the mycelial growth rate of the undesirable fungi decreased. Also, all PSA-G me-
Mycelial interaction
dia, especially PSA-G0.2, indicated low growth averages compared with the control (fresh PSA) against the
EC
pathogen while this concentration encourages growth of oyster mushroom. Also, this concentration reduced
the density of sporulation of green molds; therefore, this concentration can be applied to reduce influence this
pathogen in cultivation farm.
© 2017.

1. Introduction dual cultures. In contrast, metabolites of P. ostreatus were not stim-

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ulated growth of T. harzianum [8]. Also, T. harzianum has a highly

The fungus Pleurotus ostreatus, oyster mushroom, is one of the
popular macro-fungi which belong to higher Basidiomycetes. It is
considered good food and drug sources as antioxidant, antibacterial
and antifungal bio-factors [1]. In Iraq, Pleurotus ostreatus can be pro-
duced and grew on different cellulosic substrate sources such as wheat
straw, cardboard wastes [2] and date-palm residues successfully [3].
CO
aggressive toward A. bisporus, P. ostreatus and L. edodes that regard-
ing contamination problem which leads to a decrease in the gross pro-
duction and the financial loss for producers [5]. From the incubation
temperature profiles of the pathogen and oyster mushroom, no range
was found that would allow optimal growth of the mushrooms without
mold contamination [9].

In commercial mushroom farms, Trichoderma harzianum, T. lon-
gibrachiatum, and T. atroviride cause green molds [4]. T. harzianum,
the most dominant contaminant, is leading to green mold disease and
causing a high loss in Agaricus bisporus, P. ostreatus and Lentinula
edodes production [5]. Also, Trichoderma spp. is a common contami-
nant of spawn, compost, and wood in mushroom cultivation farm [6].
Generally, in dual culture test, T. harzianum was antagonistic to Pleu-
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Licorice or liquorice plant, Glycyrrhiza sp., is one of the medi-
cinal plants from 4000 years ago, which used as a drug in different
fields. It belongs to the family Fabaceae [10]. The primary active con-
stituent of Glycyrrhiza sp. is Glycyrrhizin known as glycyrrhetinic
acid or glycyrrhizic acid. It is fifty times sweeter than sugar (sucrose)
with concentration 6–14% [11]. From other side, Glycyrrhiza sp. is
possible to secrete flavonoids, coumarins [12], triterpenoids (Gly-

rotus spp. [7]. Whereas, mycelia of oyster mushrooms can be cover
the T. harzianum colonies when supporting the medium using oils of
tea tree Melaleuca alternifolica [6].
T. harzianum overgrew the colony of P. ostreatus rapidly due to
metabolites of T. harzianum which reduced P. ostreatus growth in
⁎ hibitory effect toward microbes. Also, using licorice extract spray on
Corresponding author at: Department of Heet Education, General Directorate of
Education in Anbar, Ministry of Education, Hit, Anbar 31007, Iraq.
Email address: mustafanowaid@gmail.com (M.N. Owaid)
cyrrhizin and volatiles) [13], glabrol (isoflavonoids) [14], tannins [15],
licoricone, kumatakenin, chalcones and phytosterols, [11]. Many stud-
ies made up toward using licorice extract to encourage the growth
of some plants and mushrooms because of naturally occurring com-
pounds [16–18]. Accordingly, Friis-Moller et al. [19] tested licochal-
cone A from licorice root powder toward anti-mycobacterial activity.
Musa et al. [15] referred to Glycyrrhizin (glycyrrhizic) have an in-
pomegranate fruits lead to determine findings of microbes [20].

https://doi.org/10.1016/j.chnaes.2017.08.001
1872-2032/© 2017.
2 Acta Ecologica Sinica xxx (2017) xxx-xxx

Strategies of possible management included the application of experiment was carried out in three replicates. The percent inhibition
chemical treatments like adjustment of pH [21], disinfectant, pasteur- was calculated using these equations:
ization and biological control using antagonistic bacteria or using re-
sistant mushroom varieties [22]. Thus this work dealt a new strategy
(using Glycyrrhiza glabra licorice root extract) to inhibit and/or re-

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duce the growth of those fungal pathogens and encourage the growth
of P. ostreatus oyster mushroom at the same time in alone and dual
cultures.

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2. Materials and methods

2.1. Fungal strains

2.6. Sporulation of green molds
Three varieties of genus Trichoderma viz.; Trichoderma spp. ob-
tained from Department of Horticulture, College of Agriculture; T. There are three levels of sporulation density, which were observed

harzianum I and T. harzianum II obtained from fungi and plant pathol-
ogy Lab. Department of Biology in University of Anbar, Iraq. White
oyster mushroom Pleurotus ostreatus obtained from MushroomBox,
UK which used for the mycelial interaction test.
2.2. Licorice root extract preparation
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on plates in case of alone and dual cultures as mentioned by [23]:
(+++): growth and sporulation similar to those of the wild-type strain
(++): reduced growth or/and sporulation
(+): highly reduced growth or/and sporulation
(−): no sporulation formation.

Licorice (Glycyrrhiza glabra) extract was prepared using topical 2.7. Statistical analysis

licorice root powder with distilled water at ratio (1:10) (w/v), left for
24 h in shaker incubator at 25 °C. It was filtrated using Whatman No.
1, centrifuged at 3000 cycle/min for 10 min and micro-filtrated us-
ing Millipore 0.22 μ [17]. The treatments were three concentrations
of G. glabra 0.05, 0.10 and 0.2 g/L (as symbolized by PSA-G0.05, TE
PSA-G0.1, and PSA-G0.2 respectively) which used in this test in com-
parison with the control (PSA).
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Statistical significance was determined by using CRD at two-way
analysis of variance (ANOVA) by implementing GenStat Discovery
Edition computer program version 7 DE3 (VSN International Ltd.,
UK). Significant differences at p < 0.05 were considered. All the ex-
periments were done in three replicates.

2.3. PSA medium supplemented with licorice extract
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Potato sucrose agar (PSA) medium was prepared as mentioned by
Owaid [17]. The prepared PSA medium was autoclaved at 121 °C
for 25 min and then poured into 85 mm Petri dishes. Before solid-
ity PSA, three concentrations of the plant extract were added to yield
PSA-G0.05 (0.05 g/L), PSA-G0.1 (0.10 g/L) and PSA-G0.2 (0.20 g/
L) [17]. Fresh PSA was considered as control.
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3. Results
3.1. Radial mycelial growth of Trichoderma spp.
In alone culture test, mycelial growth rate (MGR) of Trichoderma
spp. on PSA enriched with extract of licorice (G. glabra) root in three
concentrations (0.05, 0.10 and 0.2) g/L are presented in Table 1. The
best MGR showed on fresh PSA medium for T. harzianum II and
Trichoderma spp. at averages 36.75 and 36.50 mm/day respectively,
then declined significantly (p < 0.05) to average 34.75 and 34.50 mm/

2.4. The mycelial interaction between Pleurotus ostreatus and
Trichoderma isolates
The mycelium of Pleurotus ostreatus (white) was tested for anti-
fungal activity against three pathogenic fungi for mushroom cultiva-
tion farm viz.; Trichoderma spp., T. harzianum I and T. harzianum
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day for Trichoderma spp. and T. harzianum II on PSA-G0.05 medium
(0.05 g/L) respectively. While the lower MGR was 17.75 mm/day
recorded on PSA-G0.1 (0.10 g/L), whereas, PSA-G0.2 recorded lower
MGR for T. harzianum II and Trichoderma spp. of 31.25 and
33.50 mm/day respectively. Generally, all PSA-G media indicated
low growth averages compared to the control (Table 1). Fig. 1 showed

II on PSA medium enriched with licorice extract in three concentra-
tions using dual culture method. Culture disks measuring 5 mm were
made in the Petri dishes for each pathogenic fungus. A culture plug of
seven-day-old pathogenic fungi was placed 3 cm away from the mush-
room culture plug. The plates were then incubated for seven days at
25 ± 2 °C. Plates were checked and inhibition zone was measured and
recorded in comparing with control plates [7]. The number of days
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radial mycelial growth of Trichoderma spp.
In dual culture test (Table 2), mycelial interaction of Pleurotus os-
treatus (white) antagonistic Trichoderma spp. was applied on PSA
Table 1
Mycelial growth rate of Trichoderma spp. on PSA-G media after 2 days in alone culture
(mm/day).

taken to overgrow the pathogens was recorded.

Pathogenic fungi Culture media
2.5. Percent inhibition of the mycelial growth
Fresh PSA PSA-G0.05 PSA-G0.1 PSA-G0.2

The seven-day-old cultures of three pathogenic fungi were placed T. harzianum I 20.75 19.50 17.75 18.50
in the center of plates and incubated at 25 °C for 2 days to detect in- T. harzianum II 36.75 34.50 32.00 31.25
hibitory activity by measuring the mycelial growth rate (MGR) on Trichoderma spp. 36.50 34.75 33.75 33.50
LSD p < 0.05 0.758
PSA-G media in case dual (MGRDual) or alone (MGRPSA-G) cultures
and the radial growth on fresh PSA plate as control (MGRPSA). The PSA: fresh potato sucrose agar medium, PSA-G0.05: PSA medium with licorice (G.
glabra) extract at 0.05 g/L, PSA-G0.1: PSA medium with licorice extract at 0.10 g/L,
PSA-G0.2: PSA medium with licorice extract at 0.20 g/L.
 Acta Ecologica Sinica xxx (2017) xxx-xxx 3

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Fig. 2. Mycelial growth rate of Trichoderma spp. after 2 days in general.

3.3. Inhibition percent of green molds using PSA-G media

Fig. 1. Pattern of radial mycelial growth of Trichoderma spp. on PSA media supple-
mented with licorice root extract. Legend: PSA: fresh potato sucrose agar medium,
PSA-G0.05: PSA medium with licorice (G. glabra) extract at 0.05 g/L, PSA-G0.1: PSA
medium with licorice extract at 0.10 g/L, PSA-G0.2: PSA medium with licorice extract
at 0.20 g/L.
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It is evident from the Table 3, the significant differences (p < 0.05)
were observed among the percent inhibition of various pathogenic
fungi on serious concentrations of licorice root extract on solid
medium (PSA-G medium). In significant (p < 0.05), the best mycelial
growth inhibition was recorded on PSA-G0.2 (14.97%) by T.
harzianum II, followed 14.46% and 12.93% by T. harzianum I and
T. harzianum II on PSA-G0.1 medium respectively, then reached to

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10.84% on PSA-G0.2 in case of T. harzianum I. The lower growth
Table 2 inhibition was 4.79% in case of Trichoderma spp. on PSA-G0.05.
Mycelial growth rate of Trichoderma spp. on PSA-G media after 2 days in dual culture PSA-G0.1 considered the best treatment to inhibit Trichoderma spp.
(mm/day).

Pathogenic fungi Culture media

Fresh PSA-
TE
Mean of fungi
in alone culture while other varieties are given best treatment on
PSA-G0.2.
The force of inhibition of mycelial P. ostreatus against mycelial
pathogenic fungi viz., Trichoderma spp., T. harzianum I, and T.
PSA G0.05 PSA-G0.1 PSA-G0.2 harzianum II (in dual culture) on PSA-G media after 2 days is pre-
T. harzianum I 10.25 8.75 9.17 9.50 9.41 sented in Table 4. Generally, inhibition percent of Trichoderma spp.
EC
T. harzianum II 15.50 14.33 13.75 13.33 14.22 started from 59.18%–60.16% in case of licorice extract compared
Trichoderma 14.83 14.00 13.67 14.75 14.31
spp.
LSD p < 0.05 0.890 0.445
Table 3
PSA: fresh potato sucrose agar medium, PSA-G0.05: PSA medium with licorice (G. Percent inhibition of Trichoderma spp. on PSA-G media after 2 days in alone culture.
glabra) extract at 0.05 g/L, PSA-G0.1: PSA medium with licorice extract at 0.10 g/L,
PSA-G0.2: PSA medium with licorice extract at 0.20 g/L. Pathogenic fungi Culture media
RR

Fresh PSA PSA-G0.05 PSA-G0.1 PSA-G0.2

enriched with licorice extract. Generally, according to varieties of
T. harzianum I 0.00 6.02 14.46 10.84
Trichoderma, bigger MGR was recorded 14.31 and 14.22 mm/day T. harzianum II 0.00 6.12 12.93 14.97
for Trichoderma spp. and T. harzianum II respectively, whereas, T. Trichoderma spp. 0.00 4.79 7.53 8.22
harzianum I had been less MGR at average 9.41 mm/day significantly LSD p < 0.05 0.5825
(p < 0.05). PSA: fresh potato sucrose agar, PSA-G0.05: PSA medium enriched with licorice (G.
CO

In another meaning, T. harzianum II had lower MGR 13.33 mm/ glabra) extract at 0.05 g/L, PSA-G0.1: PSA medium enriched with licorice extract at
day on PSA-G0.2 compared with 15.50 mm/day on fresh PSA (con- 0.10 g/L, PSA-G0.2: PSA medium enriched with licorice extract at 0.20 g/L.
trol). The second, Trichoderma spp. recorded 13.67 mm/day on
PSA-G0.1 compared with 14.83 mm/day on the control medium. The Table 4
last, T. harzianum I showed the lowest mycelial growth 8.75, 9.17 Percent Inhibition of Trichoderma spp. on PSA-G media after 2 days in dual culture.
and 9.50 mm/day on PSA-G0.05, PSA-G0.1 and PSA-G0.2 compared
Fungi Culture media Mean of fungi
with 10.25 mm/day on control (fresh PSA) in dual culture with P. os-
treatus.
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Fresh PSA-
PSA G0.05 PSA-G0.1 PSA-G0.2
3.2. Sensitivity of Trichoderma spp. isolates toward PSA-G media T. harzianum I 50.60 57.83 55.83 54.22 54.62
T. harzianum II 57.82 60.99 62.59 63.72 61.28
In Fig. 2, strains of Trichoderma spp. differ in sensitivity toward Trichoderma 59.36 61.64 62.55 59.59 60.78
Pleurotus ostreatus. Mycelial growth of T. harzianum I showed lower spp.
Mean of media 55.93 60.16 60.32 59.18
sensitivity in alone (5.5 mm/day) and dual cultures (5 mm/day) dur-
ing 2 days. In alone cultures, T. harzianum II and Trichoderma spp. LSD p < 0.05, fungi = 0.805, culture media = 0.929, fungi ∗ media = 1.609. PSA: fresh
showed mycelial growth about 3 mm/day, but showed approx. 4 mm/ potato sucrose agar, PSA-G0.05: PSA medium with licorice (G. glabra) extract at
0.05 g/L, PSA-G0.1: PSA medium with licorice extract at 0.10 g/L, PSA-G0.2: PSA
day in dual cultures. medium with licorice extract at 0.20 g/L.
4 Acta Ecologica Sinica xxx (2017) xxx-xxx

with 55.93% with fresh PSA (control). According to varieties of the
fungi, T. harzianum I exhibited less sensitive compared with Tri-
choderma spp. and T. harzianum II with inhibition percent 54.62%,
60.78% and 61.28% respectively. The lower significant (p < 0.05) in-
hibition achieved by T. harzianum I on fresh PSA medium (50.60%),
3.4. Sporulation patterns of pathogens
The sporulation was differed according to verities of Trichoderma
spp. which ultimately inhibited growth of P. ostreatus. Generally,
Trichoderma spp. showed higher sporulation density (Fig. 3C), fol-

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while the isolate T. harzianum II had more inhibition percent 63.72% lowed by T. harzianum I (Fig. 3A), while the Fig. 3B showed less
on PSA-G0.2 medium.

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TE
EC
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Fig. 3. Sporulation patterns of Trichoderma spp. in alone and dual cultures.

 Acta Ecologica Sinica xxx (2017) xxx-xxx 5

Table 5
Sporulation levels of Trichoderma spp.

Fungi Test Fresh PSA PSA-G0.05 PSA-G0.1 PSA-G0.2

T. harzianum I In alone +++ ++ ++ ++

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In dual +++ + + +
T. harzianum II In alone − − − −
In dual − − − −
Trichoderma spp. In alone +++ +++ +++ +++

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In dual +++ ++ ++ ++

Table 6
Number of overgrowth days of Trichoderma spp. on PSA-G media in alone culture.

Fungi Culture media Mean of fungi

Fresh PSA-

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PSA G0.05 PSA-G0.1 PSA-G0.2
against these pathogens. Table 5 showed various degrees of sporu-
T. harzianum I 5 6 6 5 5.5 lation, which decreased with dual cultures for T. harzianum I and
T. harzianum II 3 3 3 3 3.0 Trichoderma spp. in the supplemented media with plant extract. T.
Trichoderma 3 3 3 3 3.0
spp.
harzianum II have not affected.
LSD p < 0.05 0.00 0.00 Legend: A: T. harziunam I, B: T. harziunam II, C: Trichoderma
spp.
PSA: fresh potato sucrose agar, PSA-G0.05: PSA medium with licorice (G. glabra)
extract at 0.05 g/L, PSA-G0.1: PSA medium with licorice extract at 0.10 g/L,
3.5. Comparison of overgrowth of green molds in alone and dual

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PSA-G0.2: PSA medium with licorice extract at 0.20 g/L.
culture tests on PSA-G media
Table 7
Number of overgrowth days of Trichoderma spp. on PSA-G media in dual culture. In alone culture test, the longer time used up to overgrow isolates

Fungi Culture media

Fresh
PSA
PSA-
G0.05 PSA-G0.1 PSA-G0.2
TE
Mean of fungi
of Trichoderma spp. on PSA-G media was 6 days for T. harzianum I
on PSA-G0.05 and PSA-G0.1 media significantly (p < 0.05), then de-
creased to 5 days by the same fungus on PSA-G0.2 and fresh PSA me-
dia as well. While other isolates T. harzianum II and Trichoderma spp.
had only 3 days to overgrow on all PSA-G media as seen in Table 6.
T. harzianum I 5.0 5.0 5.0 5.0 5.0
T. harzianum II 3.0 3.3 3.6 4.0 3.5
In Table 7, in dual culture, overgrowth of T. harzianum I had
EC
Trichoderma 3.3 4.0 4.0 3.6 3.7 5 days compared as (approx. 6 days) in alone culture (Table 6). How-
spp. ever, T. harzianum II and Trichoderma spp. had more time for over-
Mean of media 3.7 4.1 4.2 4.2 growth when P. ostreatus was found, approx. 4 days in dual culture in
LSD p < 0.05, fungi = 0.28, culture media = 0.32, fungi ∗ media = 0.56. PSA: fresh comparison within the alone culture (3 days) on licorice extract media
potato sucrose agar, PSA-G0.05: PSA medium with licorice (G. glabra) extract at at different concentrations which appeared in Fig. 4. T. harzianum was
0.05 g/L, PSA-G0.1: PSA medium with licorice extract at 0.10 g/L, PSA-G0.2: PSA inhibited by all PSA-G media in dual culture but P. ostreatus could
medium with licorice extract at 0.20 g/L. not overgrow in presence of the fungal pathogen (Fig. 3A, B, C).
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4. Discussion

The high concentration of licorice G. glabra extract exhibited in-

hibitory effect toward oyster mushroom mycelia in this current study
because of that 1–5% sucrose favorable to growth Pleurotus ostrea-
tus but not more [24]. In opposite, using this plant extract showed
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anti-mycotic activity especially against its competitors Trichoderma

isolates that due to the phenolic compounds (flavonoids and
coumarins) of this extract [12]. While, this plant extract supports
growth of some edible mushrooms like Pleurotus ostreatus as men-
tioned by Abdulhadi [16] who used licorice extract to support produc-
tivity and medicinal value of P. ostreatus, therefore; it can be used
to encourage producing oyster mushroom vigorously when the patho-
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genic fungi are found.

Fig. 4. Effect treatments of licorice extract on growth completion of Trichoderma spp. From other side, secondary metabolism of P. ostreatus mycelia
(days). Legend: PSA: fresh potato sucrose agar, PSA-G0.05: PSA medium enriched was important against pathogenic fungi [25] because of its ability
with licorice (G. glabra) extract at 0.05 g/L, PSA-G0.1: PSA medium enriched with to inhibit growth of pathogenic fungi using their secondary metabo-
licorice extract at 0.10 g/L, PSA-G0.2: PSA medium enriched with licorice extract at lism by-products like triterpenoides, polysaccharides, proteins and en-
0.20 g/L.
zymes [1]. Also, the ability of oyster mushroom to produce metabolic
materials such as enzymes which give it inhibition force toward de
sporulation in the plate. From Fig. 3A and B, sporulation level de-
creased when licorice root extract increased in dual cultures. Whereas,
in alone culture, same observation was seen but in higher density
in each state, that demonstrated the action of P. ostreatus
6 Acta Ecologica Sinica xxx (2017) xxx-xxx
cay cellular walls of pathogenic fungi [26]. The difference of growth
rate of Trichoderma isolates was useful to know the contact time be-
tween the green molds and oyster mushroom in vitro according to their
inheritance characteristics.
Finally, P. ostreatus showed disability to overgrow T. harzianum.
This could be due to hyphae of this pathogen that causes cell wall lysis
at the point of interaction with oyster mushroom in Petri dish [27].
Also, Trichoderma species secrete hydrolytic enzymes including
chitinases, β-glucanases and cellulases which are kind of mycotoxins
and lyse the fungal cell walls and are thought to play a role in the my-
coparasitic activity of this fungus [28]. But that may be inhibit its bio-
chemical pathways when adding licorice extract in some concentra-
tions which did not effect on mycelia of oyster mushroom. In recent
studies, phenolic compounds showed the remarkable fungistatic effect
on the Trichoderma spp. and inhibited the host mushrooms as well,
but licorice extract may be changing role of phenolic compounds to-
ward them [9].
Inclusion, in dual culture, oyster mushroom mycelium has been
given inhibition act against pathogenic fungi when be found G. glabra
extracts as supporting natural materials in culture media especially
at concentration 0.2 g/L in this work at least. Trichoderma initially
produces a dense pure white mycelium which resembles mushroom
mycelium; therefore, they are very difficult to distinguish. Mycelial
mat on the casing layer gradually turns to a green color because of the
heavy sporulation of the causal agent producing a characteristic symp-
tom of the disease [28], but when using this extract will lead to reduc-
ing the density of sporulation/growth of the pathogen as shown in Fig.
3 and Table 6.
Uncited references
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[29–31]
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Acknowledgement
This work was supported by the Department of Biology, College
of Science, University of Anbar, Iraq. We would like to express my
sincere gratitude to the department and the university.
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References
[1] Y. Patel, R. Naraian, V.K. Singh, Medicinal properties of Pleurotus species (oys-
ter mushroom): a review, World J. Fungal Plant Biol. 3 (1) (2012) 1–12.
[2] M.N. Owaid, A.M. Abed, B.M. Nassar, Recycling cardboard wastes to produce
blue oyster mushroom Pleurotus ostreatus in Iraq, Emirates J. Food. Agric. 27
(7) (2015) 537–541.
[3] M.N. Owaid, I.A. Abed, S.S. Al-Saeedi, Using of date palm fiber mixed with
CO
other lignocelluloses toward Pleurotus ostreatus (Higher Basidiomycetes) culti-
vation, Emirates J. Food Agric. 27 (7) (2015) 556–561.
[4] L. Kredics, S. Kocsube, L. Nagy, M. Komon-Zelazowska, L. Manczinger, E.
Sajben, A. Nagy, C. Vagvolgyi, C.P. Kubicek, I.S. Druzhinina, L. Hatvani, Mol-
ecular identification of Trichoderma species associated with Pleurotus ostreatus
and natural substrates of the oyster mushroom, FEMS Microbiol. Lett.
300 (2009) 58–67.
[5] O. Romero-Arenas, M.H. Lara, M.A.D. Huato, F.D. Herna'ndez, D.A.A. Victo-
ria, The characteristics of Trichoderma harzianum as a limiting agent in edible
UN
mushrooms, Rev. Colomb. Biotecnol. 11 (2) (2009) 143–151.
[6] P. Angelini, R. Pagiotti, P. Granetti, Effect of antimicrobial activity of Melaleuca
alternifolia essential oil on antagonistic potential of Pleurotus species against
Trichoderma harzianum in dual culture, World J. Microbiol. Biotechnol.
24 (2008) 197–202.
View publication stats
[7] M.N. Owaid, S.S.S. Al Saeedi, I.A. Abed, P. Shahbazi, V. Sabaratnam, Antifun-
gal activities of some Pleurotus species (Higher Basidiomycetes), WJST 14 (3)
(2017) 215–224.
[8] R.G.U. Jayalal, N.K.B. Adikaram, Influence of Trichoderma harzianum metabo-
lites on the development of green mold disease in the oyster mushroom, Cey. J.
Sci. (Bio. Sci.) 36 (1) (2007) 53–60.
F
[9] L. Hatvani, P. Sabolic, S. Kocsube, L. Kredics, D. Czifra, C. Vagvolgyi, J.
Kaliterna, D. Ivic, E. Dermic, I. Kosalec, The first report on mushroom green
mould disease in Croatia, Arh. Hig. Rada Toksikol. 63 (2012) 481–487.
[10] M.D. Shawky, Traditional Foods in the Near East, Publication Division, FAO,
OO
Rome, Italy, 1991.
[11] A. Sehrawat, V. Kumar, Herbs and bioactive compounds in prevention and treat-
ment of hepatocellular carcinoma, in: R.R. Watson, V.R. Preedy (Eds.), Bioac-
tive Foods and Extracts, Cancer Treatment and Prevention, CRC Press, US,
2011, pp. 556–577.
[12] M. Tawata, Y. Yoda, K. Aida, H. Shinda, H. Sasaki, M. Chin, T. Onay,
Auti-platelet action of Gv-7, a3-arylcounmarin derivative purified from Gly-
cyrrhiza radix, Planta Med. 56 (1990) 259–263.
[13] C.A. Newall, L.A. Anderson, J.D. Phillipson, Herbal Medicien, The Pharmaceu-
PR
tical Press, London, 1996183–186.
[14] M.M. Cowan, Plant products as antimicrobial agents, Clin. Microbiol. Rev. 12
(4) (1999) 564–582.
[15] T.N. Musa, A.J.M. Al Hadiethy, G.A. Nassir, Study of some constituents of lo-
cal licorice roots powder (Glycyrrhiza glabra), Iraqi J. Agric. Sci. 34 (2) (2003)
19–26.
[16] A.M. Abdulhadi, Use of licorice root powder to improve yield, storage life and
medicinal properties of oyster mushroom, Iraqi J. Agric. Sci. 41 (6) (2010)
71–85.
D
[17] M.N. Owaid, Biotechnology for Local Compost Preparation Used to Produce
Mushroom Agaricus bisporus, M.Sc. Thesis, Dep. Biology, College of Science,
University of Anbar, Iraq, 2009, (129 pp.).
[18] F.H. Al-Sahaf, H.G.K. Al-Marsoumi, Effect of foliar spray of GA3, liquorice
root extract and nutrients on seed production in onion, Iraqi J. Agric. Sci. 34 (2)
(2003) 37–46.
[19] A. Friis-Moller, M. Chen, K. Fuursted, S.B. Christensen, A. Kharazmi, In vitro
antimycobacterial and antilegionella activity of licochalcone A from Chinese
licorice roots, Planta Med. 68 (5) (2002) 416–419.
[20] F.H. Al-Sahaf, M.K.M. Al-Jeboury, R.M.H. Al-Dulaimy, Effect of liquorice root
extract on pomegranate fruit cracking, Iraqi J. Agric. Sci. 33 (4) (2002) 85–90.
[21] O. Romero-Arenas, M.A.D. Huato, I.H. Trevino, J.F.C.P. Lezama, A.A. Garcia,
A.D.V. Arellano, Effect of pH on growth of the mycelium of Trichoderma viride
and Pleurotus ostreatus in solid cultivation mediums, Afric. J. Agric. Res. 7 (34)
(2012) 4724–4730.
[22] L. Kredics, L.G. Jimenez, S. Naeimi, D. Czifra, P. Urban, L. Manczinger, C.
Vagvolgyi, L. Hatvani, A challenge to mushroom growers: the green mould dis-
ease of cultivated Champignons, in: A. Mendez-Vilas (Ed.), Current Research,
Technology and Education Topics in Applied Microbiology and Microbial
Biotechnology, FORMATEX, 2010, pp. 295–305.
[23] L. Hatvani, Mushroom Pathogenic Trichoderma Species: Occurrence, Biodiver-
sity, Diagnosis and Extracellular Enzyme Production, Ph.D. dissertation Faculty
of Science and Informatics, University of Szeged, 2008108.
[24] H.T. Hoa, C.-L. Wang, The effects of temperature and nutritional conditions on
mycelium growth of two oyster mushrooms (Pleurotus ostreatus and Pleurotus
cystidiosus), Mycobiology 43 (1) (2015) 14–23.
[25] M.N. Owaid, S.S.S. Al-Saeedi, I.A.A. Al-Assaffii, Antifungal activity of culti-
vated oyster mushrooms on various agro-wastes, Summa Phytopathol. 43 (1)
(2017) 9–13.
[26] G.W. Dekan, Introduction to Modern Mycology, 2nd edition, University of
Salah ad Din. Dar Alhekma for Printing and Publishing, Erbil, Iraq,
1983130–314, (Arabic Edition).
[27] M. Muskhazli, Q.Z. Faridah, R. Salfarina, T. Nor Farizan, I. Nalisha, The evi-
dence of non n-glycan linked mannose in exochitinase 42 kDa, from Tricho-
derma harzianum BIO10671 glycosylation, Malays. J. Microbiol. 2 (2) (2006)
37–41.
[28] Y.R. Danesh, E.M. Goltapeh, H. Rohani, Identification of Trichoderma species
causing green mold in button mushroom farms, distribution and their relative
abundance, Sci. Cultiv. Edible Fungi: Mush. Sci. 15 (2) (2000) 653–659.