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Saudi Journal of Biological Sciences logoLink to Saudi Journal of Biological Sciences
. 2021 Mar 4;28(6):3294–3302. doi: 10.1016/j.sjbs.2021.02.074

Potency of plant extracts against Penicillium species isolated from different seeds and fruits in Saudi Arabia

Abd El-Rahim MA El-Samawaty a, Deiaa A El-Wakil a, Salman Alamery b, Mohamed MH Mahmoud b,⁎
PMCID: PMC8176084  PMID: 34121867

Abstract

Antifungal activity of extracts of cinnamon (Cinnamomum zeylanicum), Cloves (Syzygium aromaticum), ginger (Zingiber officinale) and turmeric (Curcuma longa) were evaluated in vitro against 17 Penicillium spp. Seed disease and rotten fruit caused by these species cause considerable loss of quality for different agricultural products. Isolates of Penicillium spp. were screened for production of patulin an important serious mycotoxin. About 70.59% of Penicillium spp. produced this toxin in concentrations ranging from 4 to 31 ppb. The response of Penicillium spp.

to plant extracts differed according to the plant extract and concentration. Cinnamon extract showed the greatest effect on P. asperosporum, P. aurintogriseum and P. brevicompactum, and cloves extract produced the greatest effect on P. chermesinum and P. duclauxii. Turmeric extract had less effect on P. duclauxii. Cloves extract was the most effective in reducing the growth of Penicillium spp. On the other hand, ginger extract with all concentrations used had less effect against most Penicillium spp in the laboratory. Plant extracts are promising as natural sources of environmentally friendly compounds in laboratory studies.

Keywords: Medicinal Plant extracts, Penicillium spp., Seed-rotting disease, Storage fungi, HPLC technique and mycotoxins

1. Introduction

Corn seeds and fruits are subject to post-harvest diseases caused by fungi during storage. These diseases cause cuts, wounds and other physical damage during harvest, packing, transport, and storage. Penicillium italicum rot disease is a devastating post-harvest disease (Abramson et al., 2009, Agrios, 2005). This disease is found in produce during cooling, storage and marketing and the disease is exacerbated by wet conditions. Fungi on fruits exhibit dark blue round areas with mature fruiting bodies surrounded by white mycelia (Al-Rahmah et al., 2013, Al-Samarrai et al., 2013). Fruiting fungi are responsible for new infection in healthy produce. Blue mold disease losses are estimated to be 10–40% (Aqil et al., 2010).

Many fruits are exposed to post-harvest diseases in the field and during storage. Post-harvest disease injury are directly related to physical damage, such as cuts and wounds, during harvest, packing, transport, and storage. Corn seeds and fruits are infected by fungi during storage. Green-mold fungi on fruit exhibit dark blue round areas with mature fruiting bodies surrounded by the growth of white fungi from P. italicum (Ayoola et al., 2008). Blue fruits infected with fungi are responsible for the new infection in healthy fruits.

Several disease management options, including chemical control sprayed on fruits to reduce pathogenic fungal infection and increase storage periods, are available (Marzoug et al., 2011). Many fruits are exposed to many post-harvest diseases caused by field and storage fungi. The injuries associated with post-harvest diseases are directly related to mechanical damage, cuts, and wounds during harvest, packing, transport, and storage. The blue rot disease is the most devastating post-harvest disease caused by the fungus Penicillium italicum (Benkeblia, 2004, Boulenouar et al., 2012, Bowers and Locke, 2000). This disease manifests in gardens during cooling, storage, and marketing and becomes more serious in wet conditions. The green-molded fungi on the fruits exhibit dark blue round areas with mature germs surrounded by the growth of white fungi from P. italicum (Bragulat et al., 2008, Chen et al., 2018). The blue fruits infected with fungi are responsible for the new infection in healthy fruits. Humidity favors the development of the disease. Losses due to Penicillium spp. rotting disease are estimated to be approximately 15%–45% (Christian, Dwivedi and Dwivedi, 2012).

Chemicals are, however, responsible for increasing risks to human health and environment, and their use leads to resistance to pesticides. Development of alternatives to fungicides is needed to help control post-harvest diseases. Biological control and adoption of natural products, including seed powders, water and alcohol extracts for many plants are possible options. Botanical extract products are environmentally friendly, inexpensive and may reduce losses by discouraging pathogen growth. Plant extracts contain active compounds that inhibit the growth of plant pathogens.

Penicillium spp. are the main cause of deterioration and decomposition of a wide range of plant products after harvest, especially fruits, such as grapes (Fki et al., 2005, Gende et al., 2008). These fungi are widespread, attacking various fruits, including grapes and especially during storage and often producing a variety of mycotoxins (Magnoli et al., 2003); (Moslem et al., 2011) . Harmful mycotoxins and carcinogenic compounds, such as citrinine, patulin, penicillic acid and other secondary metabolites are produced by Penicillium spp. (Abramson et al., 2009); (Santos et al., 2002) ; (Bragulat et al., 2008) . Effective control of fruit diseases can also be achieved through many non-chemical control strategies (Kanan and Al-Najar, 2008; (Sanzani et al., 2010). One popular non-chemical option for controlling plant diseases (Wang et al., 2004); (Soylu et al., 2005) is use of extracts and essential oils of herbaceous plants. Availability, low toxicity, and environmental friendliness make plant extracts attracted targets for investigation (Harris et al., 2001); (Fawzi et al., 2009) and (Aqil et al., 2010).

Several plant extracts possess antifungal properties and can be used to suppress decomposing fungi (Ismaiel, 2008). garlic is among the most promising natural plant materials with antifungal properties (Gende et al., 2008); Rathod et al. (2010); (Yassin et al., 2013) and Znini et al., 2011). Antifungal activity of plant extracts is noted against Penicillium spp. and other fungi, as well as reduced production of mycotoxins (Rezzi et al., 2001; Ismaiel, 2008); Taskeen et al., 2011 and (Minz et al., 2012).

The present study evaluated the efficacy of four plant extracts under laboratory conditions for antifungal activity against 17 Penicillium spp., isolated from fruits and seeds collected from Al-Riyadh markets; Saudi Arabia, and identified by the Assiut University Mycological Center, Egypt (AUMC).

2. Methods

2.1. Isolation of Penicillium spp.

Fruit and seed samples were collected from several locations (markets) in Al-Riyadh; capital of Saudi Arabia. Samples and the obtained samples were cut into small pieces, sterilized with 5% sodium hypochlorite solution for 5 min followed by washing in three changes of sterile distilled water. Samples were then dried between two filter papers for one minute. Samples were placed randomly onto potato dextrose agar (PDA) in three, 9 cm diameter, Petri dishes. Dishes were incubated at 28 °C and examined daily for seven days, after which colonies were counted. Isolates were purified either by single spore or hyphal tip methods and transferred to PDA slants. Identification of fungal isolates at the Mycological Center, Assiut University, Egypt. According to Pitt (1988), used morphological and microscopic characteristics (Table 1).

Table 1.

Isolates of Penicillium spp. analyzed in this study.

No. Penicillium Species Source Aumc No.*
1 P. asperosporum Apple 7965
2 P. aurintogriseum Peanut 5860
3 P. brevicompactum Walnut 7934
4 P. chermesinum Popcorn 5847
5 P. chrysogenum Peanut 5846
6 P. citrinum Apple 7732
7 P. duclauxii Corn 5965
8 P. expansum Grape 7576
9 P. funiculosum Corn 5966
10 P. griseofulvum Sorghum 5905
11 P. glabrum Apple 7654
12 P. implicatum Peanut 5866
13 P. olsonii Peanut 5854
14 P. oxalicum Corn 5950
15 P. puberulum Grape 7934
16 P. variabile Coffee bean 5560
17 P. verrucosum Apple 8026
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Aumc. No (Assiut University Mycological Center, Egypt).

2.2. Mycotoxins assays

Tested isolates of Penicillium spp. were grown on sterilized malt extract prepared in 100 ml flasks for 7–10 days at 27 ± 2 °C with three replicates per isolate (Yassin et al., 2010). Cultures were blended for 2 min using a high-speed homogenizer and filtered using glass filter paper. Patulin was extracted from the homogenized filtrate using acetonitrile:water (5:95 v:v) (liquid mobile phase) solution. The solvent was evaporated at 35 °C under vacuum. Dried residues containing patulin were dissolved in 1 ml of the same liquid mobile phase. This extract was passed through a 0.45 μm microfilter, and analyzed on an HPLC model PerkinElmer® Brownlee™ with a validated C18, 250 mm column. The HPLC was equipped with UV detector and compounds were detected with a UV detector at a wavelength at 280 nm. Total run time for the separation was approximately 25 min at a flow rate of 1 ml/min.

2.3. In vitro antifungal activity against 17- Penicillium spp.

Antifungal activity of four plant extracts of cinnamon (Cinnamomum zeylanicum), Cloves (Syzygium aromaticum), ginger (Zingiber officinale) and turmeric (Curcuma longa) (Table 2) were evaluated in vitro against 17 species of Penicillium. One hundred grams of plant materials were homogenized in 100 ml of distilled water (1:1W/V) for 5 min using a blender (Ismaiel, 2008). Obtained extracts were filtered through a sheath layer, and used immediately, or stored at 4 °C until use.

Table 2.

Showing the medicinal plants and their common and scientific names.

No. Common Name Scientific Name Used parts
1 Cinnamon Cinnamomum zeylanicum Powder of Cinnamon
2 Cloves Syzygium aromaticum Aromatic Flower buds
3 Ginger Zingiber officinale Turmeric Rhizomes
4 Turmeric Curcuma longa Turmeric Rhizomes
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Different volumes of crude extracts were incorporated into PDA medium just before pouring into sterilized Petri dishes to obtain extract concentrations of 5%, 10%, 15%, and 20%. Petri dishes were centrally inoculated with 2 mm fungal plugs and incubated at 28 ± 2 °C for 7–10 days. Linear growth of fungi was measured at the time when pathogenic fungi completely covered medium surface in control treatments. Percentage inhibition was calculated as:

Reductioninlineargrowth(%)=R1-R2/R1∗100

R1 = The radius of control growth

R2 = The radius of fungal inhibited growth

% of inhibition of Penicillium spp.

2.4. Statistical analysis

Analysis of variance (ANOVA) performed with the MSTAT-C statistical package (Michigan State Univ., USA) was used to calculate least significant difference (LSD) to compare means.

3. Results

3.1. Mycotoxigenicity

Isolates of Penicillium spp. were screened for patulin production; 70.59% of Penicillium spp. produced the mycotoxin in concentrations that ranged from 4 to 31 ppb (Fig. 1). P. chrysogenum displayed the highest production of patulin and P. brevicompactum the least. P. citrinum and P. oxalicum produced similar amounts.

Fig. 1.

Fig. 1

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Diagram showing the production of mycotoxin patulin by (ppb.) from 17 tested Penicillium spp. by Assiut University Mycological Center.

3.2. Antifungal activity of four plant extracts against 17 Penicillium spp.

Analysis of variance of the effects of plant extracts on the growth of Penicillium spp. showed that plant extract (P), concentrations (C) and their Interaction P × C were all highly significant sources of variation for all Penicillium spp. (Table 3). The significant interaction, P * C, indicated that the response in each species of Penicillium varied depending on plant source and concentrations.

Table 3.

ANOVA of the effects of plant extract(P). concentrations (C) and their interactions (P * C) on the linear growth of Penicillium spp.

Penicillium spp. and source of variation D.F M.S F. value P. F
1- P. asperosporum
Replication 3 10.63 1.44 0.239
plant extract(P) 3 2455.43 333.55 0.000
Concentration(C) 4 7328.21 995.49 0.000
Interaction (P * C) 12 425.79 57.84 0.000
Error 57 7.36
2- P. aurintogriseum
Replication 3 30.54 1.5 0.224
plant extract(P) 3 4771.14 234.42 0.000
Concentration(C) 4 6171.16 303.20 0.000
Interaction (P * C) 12 767.76 37.72 0.000
Error 57 20.35
3- P. brevicompactum
Replication 3 0.350 0.042 0.989
plant extract(P) 3 1619.68 192.15 0.000
Concentration(C) 4 7231.08 857.88 0.000
Interaction (P * C) 12 393.09 46.63 0.000
Error 57 8.42
4- P. chermesinum
Replication 3 2.81 0.315 0.814
plant extract(P) 3 1616.61 181.28 0.000
Concentration(C) 4 8579.18 962.03 0.000
Interaction (P * C) 12 343.15 38.48 0.000
Error 57 8.91
5- P. chrysogenum
Replication 3 16.43 1.74 0.16
plant extract(P) 3 3218.10 342.09 0.000
Concentration(C) 4 6089.76 647.36 0.000
Interaction (P * C) 12 461.66 49.07 0.000
Error 57 9.40
6- P. citrinum
Replication 3 21.54 2.30 0.086
plant extract(P) 3 912.57 97.66 0.000
Concentration(C) 4 6003.53 642.49 0.000
Interaction (P * C) 12 292.88 31.34 0.000
Error 57 9.34
7- P. duclauxii
Replication 3 5.43 0.55 0.64
plant extract(P) 3 3664.24 374.63 0.000
Concentration(C) 4 4871.03 498.01 0.000
Interaction (P * C) 12 305.46 31.23 0.000
Error 57 9.78
8- P. expansum
Replication 3 19.61 1.96 0.12
plant extract(P) 3 2902.54 291.32 0.000
Concentration(C) 4 4378.81 439.49 0.000
Interaction (P * C) 12 433.33 43.49 0.000
Error 57 9.96
9- P. funiculosum
Replication 3 6.41 0.91 0.43
plant extract(P) 3 7338.81 1048.36 0.000
Concentration(C) 4 7980.56 1140.04 0.000
Interaction (P * C) 12 580.18 82.88 0.000
Error 57 7.00
10- P. griseofulvum
Replication 3 3.23 0.36 0.78
plant extract(P) 3 3610.83 405.71 0.000
Concentration(C) 4 8481.35 952.96 0.000
Interaction (P * C) 12 282.57 31.75 0.000
Error 57 8.90
11- P. glabrum
Replication 3 41.15 5.12 0.003
plant extract(P) 3 4237.91 527.36 0.000
Concentration(C) 4 4497.48 559.66 0.000
Interaction(P * C) 12 434.86 54.11 0.000
Error 57 8.03
12- P. implicatum
Replication 3 35.68 4.63 0.006
plant extract(P) 3 1526.15 198.17 0.000
Concentration(C) 4 4128.46 536.10 0.000
Interaction (P * C) 12 128.36 16.66 0.000
Error 57 7.70
13- P. olsonii
Replication 3 44.18 4.11 0.010
plant extract(P) 3 1476.41 137.52 0.000
Concentration(C) 4 3854.73 359.04 0.000
Interaction (P * C) 12 407.07 37.91 0.000
Error 57 10.73
14- P. oxalicum
Replication 3 11.87 1.84 0.150
plant extract(P) 3 2070.91 321.10 0.000
Concentration(C) 4 4302.48 667.11 0.000
Interaction (P * C) 12 328.42 50.92 0.000
Error 57 6.44
15- P. puberulum
Replication 3 10.74 1.31 0.27
plant extract(P) 3 1273.07 156.05 0.000
Concentration(C) 4 3113.32 381.62 0.000
Interaction (P * C) 12 140.26 17.19 0.000
Error 57 8.15
16- P. variabile
Replication 3 14.58 2.42 0.075
plant extract(P) 3 4454.68 739.74 0.000
Concentration(C) 4 6957.79 1155.40 0.000
Interaction(P * C) 12 524.94 87.17 0.000
Error 57 6.02
17- P. verrucosum
Replication 3 4.55 0.74 0.50
plant extract(P) 3 4470.95 770.27 0.000
Concentration(C) 4 6709.95 1156.01 0.000
Interaction (P * C) 12 453.06 78.06 0.000
Error 57 5.80
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Effects of tested plant extracts, concentrations, and their interactions on the linear growth of P. asperosporum, P. aurintogriseum, and P. brevicompactum were recorded (Table 4). P. asperosporums shows similar responses to effects of cloves and turmeric at concentrations 10 and 15%. P. aurintogriseum shows significantly different effects of all concentrations of plant extracts except for 5%. No significant difference in effects of turmeric at concentrations 15 and 20% were observed in P. brevicompactum (see Table 5.).

Table 4.

Effects of plant extract(P) and concentrations (C) and their interactions (P * C) on the linear growth (mm) of P. asperosporum, P. aurintogriseum and P. brevicompactum.

P. asperosporum Plant Extracts
 Concentration
 Mean
Control
 5%
 10%
 15%
 20%
Cloves
 90
 85.5
 71
 63.75
 13
 64.65
Cinnamon
 90
 61.5
 53
 32.75
 20.75
 51.6
Turmeric
 90
 80.25
 73
 63.5
 46.25
 70.6
Ginger
 90
 90
 83.75
 65.5
 59.5
 77.75
Mean 90 79.31 70.19 56.37 34.87
LSD for interaction = 3.8 LSD for plant extract = 1.7 LSD for Concentration = 1.9
P. aurintogriseum Plant Extracts Control 5% 10% 15% 20% Mean
Cloves 85 70 51 33.5 11.5 50.20
Cinnamon 85 73.50 53.25 20.25 11 48.60
Turmeric 85 71.5 68 61.5 44.75 66.15
Ginger 85 84.25 81.5 79.75 77 81.50
Mean 85 74.81 63.44 48.75 36.06
LSD for interaction = 6.32 LSD for plant extract = 2.82 LSD for Concentration = 3.16
P. brevicompactum Extracts of Control 5% 10% 15% 20% Mean
Cloves 90 76 64.75 54 12.25 59.4
Cinnamon 90 72.25 54.75 44.25 22 56.65
Turmeric 90 65.75 57.75 45 42 60.1
Ginger 90 84.75 75.25 70.75 61 76.35
Mean 90 74.68 63.12 53.5 34.31
LSD for interaction = 4.06 LSD for plant extract = 1.82 LSD for Concentration = 2.03
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Table 5.

Effects of plant extract (P), concentration (C), and their interactions (P * C) on the linear growth (mm) of P. chermesinum, P. chrysogenum, and P. citrinum

P. chermesinum Plant Extracts
 Concentration
Control
 5%
 10%
 15%
 20%
 Mean
Cloves
 90
 50.5
 42.5
 40
 9
 46.40
Cinnamon
 90
 70
 50
 22.75
 17.25
 50
Turmeric
 90
 64.75
 55.5
 48.5
 45
 60.75
Ginger
 90
 70.25
 65
 58.5
 44.25
 65.6
Mean 90 63.87 53.25 42.43 28.87
LSD for interaction = 4.18 LSD for plant extract = 1.87 LSD for Concentration = 2.09
P. chrysogenum Extracts of Control 5% 10% 15% 20% Mean
Cloves 74.25 50.5 44.75 27.5 10.25 41.45
Cinnamon 74.25 64.5 29.5 20.75 9 39.6
Turmeric 74.25 71 54 50.25 47 59.3
Ginger 74.25 70.75 61.5 53.25 44.5 60.85
Mean 74.25 64.18 44.93 37.93 27.68
LSD for interaction = 4.29 LSD for plant extract = 1.92 LSD for Concentration = 2.15
P. citrinum Extracts of Control 5% 10% 15% 20% Mean
Cloves 82.75 80.5 71.25 65 16 63.1
Cinnamon 82.75 69.5 56 42.25 21.5 54.4
Turmeric 82.75 66.25 62.25 54.75 41 61.4
Ginger 82.75 79 77.75 63.75 51 70
Mean 82.75 73.81 66.81 56.43 32.37
LSD for interaction = 4.28 LSD for plant extract = 1.91 LSD for Concentration = 2.14
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Concentrations of 20% for both turmeric and ginger showed a significant and similar effect in reducing linear growth of P. chermesinum and P. chrysogenum. Further, similar inhibitory effects were found at 5% and 15% concentrations of cloves and ginger extract against P. citrinum.

No significant differences were found between the activity of cloves and ginger at concentrations 5 and 10% against P. duclauxii (Table 6). All investigated concentrations for all extracts were effective in reducing the linear growth of P. funiculosum except 5% for ginger.

Table 6.

Effects of plant extract(P). concentrations (C) and their interactions (P * C) on the linear growth (mm) of P. duclauxii, P. expansum, and P. funiculosum

P. duclauxii
Concentration
Plant Extracts
 Control
 5%
 10%
 15%
 20%
 Mean
Cloves
 81.75
 36.5
 34.75
 19.5
 13.25
 37.15
Cinnamon
 81.75
 64.25
 52.5
 47.25
 26.75
 54.5
Turmeric
 81.75
 75
 62.5
 56.75
 54.25
 66.05
Ginger
 81.75
 68.75
 67.5
 64.25
 45.5
 65.55
Mean 81.75 61.12 54.31 46.93 34.93
LSD for interaction = 4.38 LSD for plant extract = 1.96 LSD for Concentration = 2.19
P. expansum Extracts of Control 5% 10% 15% 20% Mean
Cloves 67 52.5 25.75 17 9.75 34.25
Cinnamon 67 65.75 41 34 18.5 45.25
Turmeric 67 65.75 65 44.5 36.75 55.8
Ginger 67 63.5 62.5 59.5 51.75 60.85
Mean 67 61.87 48.56 38.75 29.18
LSD for interaction = 4.42 LSD for plant extract = 1.98 LSD for Concentration = 2.21
P. funiculosum Extracts of Control 5% 10% 15% 20% Mean
Cloves 90 63.5 39 35.5 9 47.4
Cinnamon 90 41.75 30.5 21 9 38.95
Turmeric 90 78.25 68.75 56 50 68.6
Ginger 90 86.5 81.5 73.5 60 78.3
Mean 90 67.5 54.93 46 32
LSD for interaction = 3.7 LSD for plant extract = 1.66 LSD for Concentration = 1.85
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P. griseofulvum shows the same responses to effect cinnamon and turmeric at concentrations 5 and 15%, and P. implicatum shows the same response to similar concentrations of turmeric and ginger extracts (Table 7). P. glabrum shows significant responses to all extracts and all concentrations except 5 %ginger.

Table 7.

Effect of plant extract(P). concentrations (C) and their interactions (P * C) on the linear growth (mm) of P. griseofulvum, P. glabrum, and P. implicatum

P. griseofulvum
Concentration
Plant Extracts
 Control
 5%
 10%
 15%
 20%
 Mean
Cloves
 90
 41.25
 34.75
 18.25
 9
 38.65
Cinnamon
 90
 77.5
 62.5
 40.5
 32
 60.5
Turmeric
 90
 73.5
 54.25
 44.5
 40.75
 60.5
Ginger
 90
 86
 68.25
 61.25
 46.75
 70.45
Mean 90 69.56 54.93 41.12 32.12
LSD for interaction = 4.18 LSD for plant extract = 1.87 LSD for Concentration = 2.09
p. glabrum Extracts of Control 5% 10% 15% 20% Mean
Cloves 72.25 53 33.25 27.25 9 38.95
Cinnamon 72.25 53.25 31.75 14.75 9.75 36.35
Turmeric 72.25 66.75 65.5 60 56.75 64.25
Ginger 72.25 69.75 64.75 53.5 45.5 61.15
Mean 72.25 60.68 48.81 38.87 30.25
LSD for interaction = 3.97 LSD for plant extract = 1.77 LSD for Concentration = 1.98
p. implicatum Extracts of Control 5% 10% 15% 20% Mean
Cloves 67 43.5 31 22.25 9.6 34.65
Cinnamon 67 58.75 44.25 37.25 22.25 45.9
Turmeric 67 64 52.75 42 31.75 51.5
Ginger 67 63.5 54 44.25 43.5 54.45
Mean 67 57.43 45.5 36.43 26.75
LSD for interaction = 3.89 LSD for plant extract = 1.74 LSD for Concentration = 1.94
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Cloves and cinnamon extracts showed a significant effect in reducing the linear growth of P. olsonii, P. oxalicum and P. uberulum at all concentrations (Table 8). Equal effects of turmeric extracts were similar at concentrations of 10% with effects of ginger extracts at concentrations 15% against P. olsonii and P. oxalicum.

Table 8.

Effects of plant extract(P). concentrations (C) and their interactions (P * C) on the linear growth (mm) of P. olsonii, P.oxalicum, and P. uberulum.

P. olsonii
Concentration
Plant Extracts
 Control
 5%
 10%
 15%
 20%
 Mean
Cloves
 62.75
 52.25
 45.75
 36
 9.5
 41.25
Cinnamon
 62.75
 58.0
 50.75
 11
 9
 38.3
Turmeric
 62.75
 60.0
 53.5
 45.25
 32.75
 50.85
Ginger
 62.75
 60.0
 58.0
 53.5
 50.25
 56.9
Mean 62.75 57.56 52.0 36.43 25.37
LSD for interaction = 4.59 LSD for plant extract = 2.05 LSD for Concentration = 2.29
P. oxalicum Extracts of Control 5% 10% 15% 20% Mean
Cloves 58.75 46.25 11.75 9 9 26.95
Cinnamon 58.75 52.75 33 11 9 32.9
Turmeric 58.75 51.75 45 40.25 29.5 45.05
Ginger 58.75 55 51.75 45 32.75 48.65
Mean 58.75 51.43 35.37 26.31 20.06
LSD for interaction = 3.56 LSD for plant extract = 1.59 LSD for Concentration = 1.78
P. uberulum Extracts of Control 5% 10% 15% 20% Mean
Cloves 63.5 46.5 27.75 22 9.75 33.9
Cinnamon 63.5 47.75 37.75 32.75 30.5 42.45
Turmeric 63.5 43.75 35.75 34 27.5 40.90
Ginger 63.5 57.75 51.75 49.25 43.75 53.2
Mean 63.5 48.93 38.25 34.5 27.87
LSD for interaction = 4.0 LSD for plant extract = 1.79 LSD for Concentration = 2.0
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Cinnamon at concentrations 10% and turmeric at concentrations 15% showed similar impacts against P. variabile (Table 9). Turmeric at concentrations of 20% and ginger at concentrations of 15% also show similar impacts on P. verrucosum.

Table 9.

Effects of plant extract(P). concentrations (C) and their interactions (P * C) on the linear growth (mm) of P. variabile and P. verrucosum.

P. variabile
Concentration
Plant Extracts
 Control
 5%
 10%
 15%
 20%
 Mean
Cloves
 90
 60.75
 57.75
 44.5
 9.5
 52.5
Cinnamon
 90
 71
 61.75
 52.75
 9
 56.9
Turmeric
 90
 75.25
 68
 61.75
 41.25
 67.25
Ginger
 90
 90
 88.75
 88
 73.5
 86.05
Mean 90 74.25 69.06 61.75 33.31
LSD for interaction = 3.44 LSD for plant extract = 1.54 LSD for Concentration = 1.72
P. verrucosum Extracts of Control 5% 10% 15% 20% Mean
Cloves 90 56.25 45.5 31.5 9.75 46.6
Cinnamon 90 77.5 60.75 40.75 19.25 57.65
Turmeric 90 90 86.5 76.25 63.25 81.20
Ginger 90 75.25 66.5 63.25 53.25 69.65
Mean 90 74.75 64.81 52.93 36.37
LSD for interaction = 3.37 LSD for plant extract = 1.51 LSD for Concentration = 1.69
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ANOVA (Table 10) for linear growth (mm) of Penicillium spp. demonstrated highly significant impacts of plant extracts (p = 0.000). LSD was calculated to compare Penicillium ssp. mean growth for each plant extract.

Table 10.

ANOVA of the effects of plant extract (P), Penicillium spp. (P.S) and their interactions (P.S * P) on the linear growth (mm) of Penicillium spp.

Source of variation D.F M.S F.value P F
Replication 3 23964.11 257.21 0.000
plant extract(P) 3 9760.17 104.76 0.000
Penicillium spp. (P.S) 4 1445.22 15.51 0.000
Interaction (P.S * P) 12 196.33 2.10 0.000
Error 57 93.16
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Responses of P. brevicompactum, P. chermesinum, and P. griseofulvum to turmeric extract are almost equal but responses of other species it responded to the other extracts were significantly different. P. implicatum and P. olsonii showed significant response to all extracts except ginger extract. On the other hand, P. funiculosum and P. variabile show significant response to all extracts except turmeric extract. (Table 11)

Table 11.

Effects of plant extract(P). Penicillium spp. (P.S) and their interactions (P.S * P) on the linear growth (mm) of Penicillium spp.

Penicillium spp. Plant Extracts
 Mean
Control Cloves Cinnamon Turmeric Ginger
1 P. asperosporum 90.00 64.65 51.6 70.60 77.75 66.15
2 P. aurintogriseum 85.00 50.20 48.6 66.15 81.50 61.61
3 P. brevicompactum 90.00 59.00 56.65 60.10 76.35 63.02
4 P. chermesinum 90.00 46.40 50.00 60.75 65.60 55.68
5 P. chrysogenum 74.25 41.45 35.60 59.30 60.85 49.30
6 P. citrinum 82.75 63.10 54.4 61.40 70.85 62.43
7 P. duclauxii 81.75 37.15 54.5 66.05 65.55 55.81
8 P. expansum 67.00 34.25 43.70 55.80 60.85 48.65
9 P. funiculosum 90.00 47.40 36.45 68.6 78.30 57.68
10 P. griseofulvum 90.00 38.65 60.50 60.40 70.45 57.5
11 P. glabrum 72.25 38.95 36.35 64.25 61.15 50.17
12 P. implicatum 67.00 34.65 45.9 51.50 54.45 46.62
13 P. olsonii 62.75 41.25 38.3 50.85 56.90 46.82
14 P. oxalicum 58.75 26.95 32.9 45.05 48.65 38.38
15 P. puberulum 63.5 33.75 42.45 40.90 53.20 42.57
16 P. variabile 90.00 52.50 56.90 67.25 86.05 65.67
17 P. verrucosum 90.00 46.60 57.65 81.20 69.65 63.77
Mean 79.11 44.52 47.20 60.59 66.94
LSD for interaction = 2.98 LSD for plant extract = 1.45 LSD for Concentration = 1.66
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A phenogram based on average linkage cluster analysis of the response of Penicillium spp. to different plant extracts shows three distinct groups of isolated Penicillium spp. (Fig. 2) Each is divided into two subgroups; strongly and positively associated Penicillium spp. were grouped in the same cluster. The grouping pattern of the Penicillium spp. in the cluster analysis did depend on the source of the Penicillium isolate.

Fig. 2.

Fig. 2

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Phenogram based on average linkage cluster analysis of the response of Penicillium spp. to plant extracts.

4. Discussion

Different strategies are employed for controlling a serious plant pathogenic fungi worldwide; one important approach is employing plant extracts. Such extracts are considered safe and effective alternatives (Al-Rahmah et al., 2013) and (Al-Samarrai et al., 2013); (Aqil et al., 2010); (El-Samawaty et al., 2013) and (Abramson et al., 2009). Four plant extracts showed significant variation for inhibition of mycelial growth for all the investigated Penicillium spp. in vitro. Production of mycotoxins also fluctuated among Penicillium spp.

Isolates of Penicillium spp. were screened for production of the mycotoxin, patulin; 70.59% of species produced patulin in varying amounts depending on species. These results are consistent with (Yassin et al., 2010) and (Moslem et al., 2011) who investigated fungal ochratoxin production on different plant materials and suggested this toxin as an important factors for reducing self-life in Saudi Arabia.

Antifungal activity of four plant extracts against 17 Penicillium spp. showed that plant extract, concentrations and their interaction were all highly significant sources of variation in the inhibition of examined species. The significant interaction of extract and concentrations indicated that both factors contributed to variation in Penicillium spp. test. Earlier workers investigated effects of different plant extracts on controlling pathogenic fungi and observed that concentrations of extracts is a critical factor for reduction in mycelia growth (Wang et al., 2004; Soylu et al., 2005; Ismaiel, 2008 and Taskeen-Un-Nisa and Mir, 2010).

The activity of cinnamon (C. zeylanicum) extract against penicillium spp. could be attributed to the presence of Cinnamaldehyde, eugenol and cinamic acid in addition to flavonoids, alkaloiks, tannins and saponins suggested by some investigators as antifungal agents. Mahmoud (2012). Clove (S. aromaticum) extract also found to be very active against the tested penicillium spp. This activity could be attributed to the presence of phenolic compounds such as eugenol are highly active against microorganisms. Laila Muñoz Castellanos et al. (2020). Phenolic compounds such as gingerol, cedrene, zingiberene in ginger (Z. officinale) extract were determined as the most effective antifungal components; which play the vital role in growth inhibition of phytopathogenic fungi (Mostafa et al., 2011); (Al-Rahmah et al., 2013). Chen et al., 2018. They found the following compounds curdione, isocurcumenol, curcumenol, curzerene, β-elemene, curcumin, germacrone, curcumol in the extract of Turmeric (Curcuma longa). Which were effective against Penicillium pallidum and other fungi.

Analysis of variance for linear growth (mm) of Penicillium spp. showed highly significant impacts from exposure to plant extracts. These findings are consistent with results of Bowers et al., 2000; Obagwu and Korsten, 2003; Dwivedi, et al., 2012; (El-Samawaty et al., 2013) and (Al-Rahmah et al., 2013). Further, a phenogram based on average linkage cluster shows three distinct groups of isolated Penicillium spp. with strongly and positively associated Penicillium spp. grouped into the same cluster. The grouping pattern is similar to observed by earlier workers (Omar et al., 2007) and (Peng et al., 2012)who indicated that geographical origin didn’t correlate with the source of isolated fungi and variations in results of grouping may due to genetic variation among isolates.

5. Conclusion

The present study shows the natural and ecological diversity of plants with anti-microbial activity. Comprehensive explorations are needed to identify more plants with these properties. Active compounds can then be identified, formulated and made available to farmers for use as pesticides to reduce the harmful effects of using fungicides.

Author contributions

A M E and D A E carried out isolation and mycotoxin analysis. A M E, SA, and MHM designed the study, performed the statistical analysis, and participated in the manuscript drafting.

Funding

This work was supported by Researchers Supporting Project number RSP-2020/241, King Saud University.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

Acknowledgments

Authors of the present study hope to introduce their deepest thanks to all staff members in Assiut University Mycological Center (AUMC), Egypt for identification the Penicillium spp. And putting the isolates in serial code numbers in the present study and for their continuous encouragements. The authors would like to extend their gratitude to King Saud University, Riyadh, Saudi Arabia, for the funding of this research through Researchers Supporting Project number RSP-2020/241.

Ethics approval and consent to participate: Not applicable.

Consent for publication: Not applicable.

Availability of data and materials: Not applicable.

Footnotes

Peer review under responsibility of King Saud University.

Contributor Information

Abd El-Rahim M.A. El-Samawaty, Email: aelsamawaty@gmail.com.

Deiaa A. El-Wakil, Email: de107@yahoo.com.

Salman Alamery, Email: salamery@KSU.EDU.SA.

Mohamed M.H. Mahmoud, Email: mmahmoud2@ksu.edu.sa.

References

  1. Abramson D., Lombaert G., Clear R.M., Sholberg P., Trelka R., Rosin E. Production of patulin and citrinin by Penicillium expansum from British Columbia (Canada) apples. Mycotoxin Res. 2009;25(2):85–88. doi: 10.1007/s12550-009-0012-4. [DOI] [PubMed] [Google Scholar]
  2. Agrios G.N. 5th Edition. Elsevier Academic Press; Burlington, Ma. USA: 2005. Plant pathology; pp. 79–103. [Google Scholar]
  3. Al-Rahmah A.N., Mostafa A.A., Abdel-Megeed A., Yakout S.M., Hussein S.A. Fungicidal activities of certain methanolic plant extracts against tomato phytopathogenic fungi. African J. Microbiol. Res. 2013;7(6):517–524. [Google Scholar]
  4. Al-Samarrai G.F., Singh H., Syarhabil M. Extracts some plants on controlling green mold of orange and on postharvest quality parameters. World Appl. Sci. J., Deira. 2013;22(4):564–570. [Google Scholar]
  5. Aqil, F., Zahin, M., Ahmad, I., Owais, M., Khan, M.S., Bansal, S.S., Farooq, S., 2010. Antifungal activity of medicinal plant extracts and phytocompounds: A review. InCombating Fungal Infections 2010, pp. 449–484. Springer, Berlin, Heidelberg.
  6. Ayoola G.A., Lawore F.M., Adelowotan T., Aibinu I.E., Adenipekun E., Coker H.A., Odugbemi T.O. Chemical analysis and antimicrobial activity of the essential oil of Syzigiu aromaticum (clove) African J. Microbiol. Res. 2008;2(7):162–166. [Google Scholar]
  7. Marzoug H.N., Romdhane M., Lebrihi A., Mathieu F., Couderc F., Abderraba M., Khouja M.L., Bouajila J. Eucalyptus oleosa essential oils: chemical composition and antimicrobial and antioxidant activities of the oils from different plant parts (stems, leaves, flowers and fruits) Molecules. 2011;16(2):1695–1709. doi: 10.3390/molecules16021695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Benkeblia N. Antimicrobial activity of essential oil extracts of various onions (Allium cepa) and garlic (Allium sativum) LWT-food Sci. Technol. 2004;37(2):263–268. [Google Scholar]
  9. Boulenouar N., Marouf A., Cheriti A., Belboukhari N. Medicinal Plants Extracts as Source of Antifungal Agents against Fusarium oxysporum f. sp. albedinis. J. Agr. Sci. Tech. 2012;14:659–669. [Google Scholar]
  10. Bowers J.H., Locke J.C. Effect of botanical extracts on the population density of Fusarium oxysporum in soil and control of Fusarium wilt in the greenhouse. Plant Dis. 2000;84(3):300–305. doi: 10.1094/PDIS.2000.84.3.300. [DOI] [PubMed] [Google Scholar]
  11. Bragulat M.R., Abarca M.L., Cabanes F.J. Low occurrence of patulin-and citrinin-producing species isolated from grapes. Lett. Appl. Microbiol. 2008;47(4):286–289. doi: 10.1111/j.1472-765x.2008.02422.x. [DOI] [PubMed] [Google Scholar]
  12. Chen C., Long L., Zhang F., Chen Q., Chen C., Yu X. Antifungal activity, main active components and mechanism of Curcuma longa extract against Fusarium graminearum. PLoS ONE. 2018;13(3) doi: 10.1371/journal.pone.0194284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Christian, G. HPLC tips and tricks. Great Britain at the Iden Press.
  14. Dwivedi S.K., Dwivedi N. Antifungal activity of some plant extracts against guava wilt pathogen. Int. J. Environ. Sci. 2012;3(1):412–420. [Google Scholar]
  15. El-Samawaty A.E., Yassin M.A., Moslem M.A., Omar M.R. Effectiveness of Some Plant Extracts against Fusarium spp Causing Cotton Seedlings Damping-off. Life Sci. J. 2013;10(4):510–515. [Google Scholar]
  16. Fawzi E.M., Khalil A.A., Afifi A.F. Antifungal effect of some plant extracts on Alternaria alternata and Fusarium oxysporum. Afr. J. Biotechnol. 2009;8(11) [Google Scholar]
  17. Fki I., Allouche N., Sayadi S. The use of polyphenolic extract, purified hydroxytyrosol and 3, 4-dihydroxyphenyl acetic acid from olive mill wastewater for the stabilization of refined oils: a potential alternative to synthetic antioxidants. Food Chem. 2005;93(2):197–204. [Google Scholar]
  18. Gende L.B., Floris I., Fritz R., Eguaras M.J. Antimicrobial activity of cinnamon (Cinnamomum zeylanicum) essential oil and its main components against Paenibacillus larvae from Argentine. Bull. Insectol. 2008;61(1):1. [Google Scholar]
  19. Harris J.C., Cottrell S.L., Plummer S., Lloyd D. Antimicrobial properties of Allium sativum (garlic) Appl. Microbiol. Biotechnol. 2001;57(3):282–286. doi: 10.1007/s002530100722. [DOI] [PubMed] [Google Scholar]
  20. Ismaiel A.A. Third Environmental Conference Faculty of Science. Zagazig University; 2008. Inhibitory effect of garlic extract of penicillic acid production and growth of Penicillium hirsutum; pp. 45–58. [Google Scholar]
  21. Kanan G.J., Al-Najar R.A. In vitro antifungal activities of various plant crude extracts and fractions against citrus post-harvest disease agent Penicillium digitatum. Jordan Journal of Biological Sciences. 2008;1(3):89–99. [Google Scholar]
  22. Laila Muñoz Castellanos, Nubia Amaya Olivas , Juan Ayala-Soto, Carmen Miriam De La O Contreras, Miriam Zermeño Ortega, Fabiola Sandoval Salas, and Leon Hernández-Ochoa, 2020. In Vitro and In Vivo Antifungal Activity of Clove (Eugenia caryophyllata) and Pepper (Piper nigrum L.) Essential Oils and Functional Extracts Against Fusarium oxysporum and Aspergillus niger in Tomato (Solanum lycopersicum L.) International Journal of Microbiology, Article ID 1702037, 8 pages. [DOI] [PMC free article] [PubMed]
  23. Magnoli C., Violante M., Combina M., Palacio G., Dalcero A. Mycoflora and ochratoxin-producing strains of Aspergillus section Nigri in wine grapes in Argentina. Lett. Appl. Microbiol. 2003;37(2):179–184. doi: 10.1046/j.1472-765x.2003.01376.x. [DOI] [PubMed] [Google Scholar]
  24. Minz S., Samuel C.O., Tripathi S.C. The effect of plant extracts on the growth of wilt causing fungi Fusarium oxysporum. J. Pharm. Biol. Sci. 2012;4(1):13–16. [Google Scholar]
  25. Moslem M.A., Yassin M.A., El-Samawaty A.M., Sayed S.R. New toxigenic Penicillium species associated with apple blue mold in Saudi Arabia. Fresenius Env. Bull. 2011;20:3194–3198. [Google Scholar]
  26. Mostafa A.A., Al-Rahmah A.N., Adel-Megeed A. Evaluation of some plant extracts for their antifungal and antiaflatoxigenic activities. J. Med. Plant. Res. 2011;517:4231–4238. [Google Scholar]
  27. Obagwu J., Korsten L. Integrated control of citrus green and blue molds using Bacillus subtilis in combination with sodium bicarbonate or hot water. Postharvest Biol. Technol. 2003;28(1):187–194. [Google Scholar]
  28. Omar M.R., El-Samawaty A.M., El-Wakil D.A. Suppression of Pythium ultimum involved in cotton seedling damping-off by Trichoderma spp. Egypt. J. Phytopath. 2007;35(2):111–124. [Google Scholar]
  29. Peng L., Yang S., Cheng Y.J., Chen F., Pan S., Fan G. Antifungal activity and action mode of pinocembrin from propolis against Penicillium italicum. Food Sci. Biotechnol. 2012;21(6):1533–1539. [Google Scholar]
  30. Pitt JI, Hocking AD. A laboratory guide to common Penicillium species. North Ryde, NSW Australia: CSIRO Food Research Laboratory; 1988.
  31. Rathod L.R., Jadhav M.D., Awate M.K., Surywanshi A.M., Deshmukh P.S. Utilization of medicinal plants to control seed borne pathogens of selected legumes seeds. Inter. J. Advan. Biotechnol. and Res. 2010;1(2):57–59. [Google Scholar]
  32. Rezzi S., Cavaleiro C., Bighelli A., Salgueiro L., da Cunha A.P., Casanova J. Intraspecific chemical variability of the leaf essential oil of Juniperus phoenicea subsp. turbinata from Corsica. Biochem. Syst. Ecol. 2001 Feb 1;29(2):179–188. doi: 10.1016/s0305-1978(00)00044-2. [DOI] [PubMed] [Google Scholar]
  33. Mahmoud Sameer N. Antifungal Activity of Cinnamomum zeylanicum and Eucalyptus microtheca Crude Extracts Against Food Spoilage Fungi. Euphrates. J. Agric. Sci. 2012;4(3):26–39. [Google Scholar]
  34. Santos I.M., Abrunhosa L., Venâncio A., Lima N. The effect of culture preservation techniques on patulin and citrinin production by Penicillium expansum Link. Lett. Appl. Microbiol. 2002;35(4):272–275. doi: 10.1046/j.1472-765x.2002.01186.x. [DOI] [PubMed] [Google Scholar]
  35. Sanzani S.M., Schena L., De Girolamo A., Ippolito A., González-Candelas L. Characterization of genes associated with induced resistance against Penicillium expansum in apple fruit treated with quercetin. Postharvest Biol. Technol. 2010;56(1):1. [Google Scholar]
  36. Soylu E.M., Tok F.M., Soylu S., Kaya A.D., Evrendilek G.A. Antifungal activities of the essential oils on post-harvest disease agent Penicillium digitatum. Pak. J. Biol. Sci. 2005;8(1):25–29. [Google Scholar]
  37. Taskeen-Un-Nisa W.A., Mir R.A. Antimycotic activity of plant extracts on the spore germination of some pathogenic fungi. Mycopathology. 2010;8:65–69. [Google Scholar]
  38. Wang W., Ben-Daniel B.H., Cohen Y. Control of plant diseases by extracts of Inula viscosa. Phytopathology. 2004;94(10):1042–1047. doi: 10.1094/PHYTO.2004.94.10.1042. [DOI] [PubMed] [Google Scholar]
  39. Yassin M.A., El-Samawaty A.R., Bahkali A., Moslem M., Abd-Elsalam K.A., Hyde K.D. Mycotoxin-producing fungi occurring in sorghum grains from Saudi Arabia. Fungal Diversity. 2010;44(1):45–52. [Google Scholar]
  40. Yassin M.A., Moslem M.A., El-Samawaty A.E., El-Shikh M.S. Effectiveness of Allium sativum in controlling sorghum grain molding fungi. J. Pure Appl. Microbiol. 2013;7(1):101–107. [Google Scholar]
  41. Znini M., Cristofari G., Majidi L., Mazouz H.A., Tomi P., Paolini J.U., Costa J. Antifungal activity of essential oil from Asteriscus graveolens against postharvest phytopathogenic fungi in apples. Nat. Prod. Commun. 2011 Nov;6(11):1763–1768. [PubMed] [Google Scholar]

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