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Evaluation of the efficiency of Trichoderma, Penicillium, and Aspergillus species as biological control agents against four soil-borne fungi of melon and watermelon

Egyptian Journal of Biological Pest Control201828:25

https://doi.org/10.1186/s41938-017-0010-3

Received: 12 July 2017

Accepted: 6 December 2017

Published: 15 March 2018

Abstract

Various experiments were carried out to promote biological control under semi-arid ecological conditions. In vitro assay, Aspergillus flavus seemed to be the most effective bioagent against Fusarium oxysporum f. sp. niveum and Fusarium solani f. sp. cucurbitae with mycelial inhibition rate above 50%. Aspergillus flavus, Aspergillus niger, and Aspergillus terreus exhibited an exceptional hyperparasitism against F. oxysporum f. sp. melonis. The mycelial growth of five Macrophomina phaseolina isolates decreased in the presence of Trichoderma harzianum (44.42%). In greenhouse experiments, both A. flavus and A. fumigatus used preventively of melon inoculated with FOM generated the highest damage reduction rate of top and root dry weights (40–42 and 51–52%, respectively) and the lowest disease severity index (DSI). A. flavus was also effective in improving the plant development with the highest shoot (SDW) and root (RDW) dry weight values. Penicillium digitatum, Trichoderma harzianum, and Trichoderma viride treated preventively on watermelon and melon plants inoculated with M. phaseolina decreased the damage of SDW and RDW and DSI. The development rate revealed the growth improvement potential of T. harzianum (watermelon, 15%) and A. flavus (melon, 12%). Watermelon plants inoculated with F. solani f. sp. cucurbitae and treated curatively with Trichoderma erinaceum, T. viride, and A. flavus and other inoculated by F. oxysporum f. sp. niveum and treated by Trichoderma helicum recorded the highest values of growth parameters, similarly for T. erinaceum on melon plants inoculated by F. solani f. sp. cucurbitae. Among all treatments for plants inoculated by F. oxysporum f. sp. melonis, those three bioagents T. viride, T. erinaceum, and A. flavus revealed efficiency in plant growth. Trichoderma harzianum is the best bioagents against cucurbit soil-borne pathogens. Preventive treatment represents an effective strategy. Dipping roots with bioagent fungi suspension improve a good interaction pathogen antagonist.

Keywords

Root pathogensBiological control activity Fusarium species Macrophomina phaseolina MelonWatermelon

Introduction

Fusarium species are worldwide pathogenic fungi of many crop plants. Fusarium oxysporum Schltdl. is one of the most important phytopathogens causing Fusarium wilt disease in more than a hundred species of plants (Boughalleb & El Mahjoub 2006). The disease management of Fusarium wilt usually consists of soil fumigation, seed treatment, use of disease resistant varieties, and biological control bacteria to reduce infection and disease severity (Zhang et al. 2008). Fusarium root and stem rot are regarded as also one of the most devastating diseases in cucurbits (Pavlou & Vakalounakis 2005). Due to the persistent nature of these pathogens in soil, subsequent crops of susceptible melon and watermelon cultivars increase pathogen populations. The diseases are best managed with resistant cultivars. However, new virulent populations (physiological races) may develop in specific locations. Biological control of soil-borne pathogens by microorganisms has been considered a good environmentally alternative to the chemical treatment methods (Eziashi et al. 2007). Many antagonistic microorganisms have been proved to be active In vitro or in vivo. Trichoderma spp. Pers. (Shabir-U-Rehman, et al. 2013), Aspergillus species Micheli (Suárez-Estrella et al. 2007), and Penicillium spp. Link (De Cal et al. 2009) are the most known among the extensive lists. Trichoderma spp. are the most widely studied biological control agents (BCAs) for root and shoot pathogens (Hajieghrari et al. 2008), applied even in post-harvest (Woo et al. 2014). Gava and Menezes (2012) showed that selected isolates of Trichoderma spp. were efficient to control soil-borne pathogens of melon in field. Several microorganisms have been reported as plant pathogen antagonists, but only a small number were applied on a commercial scale (Fravel 2005).

The use of rhizophere Trichoderma harzianum Rifai, for controlling the spread of Macrophomina phaseolina in agronomical crops, has been suggested (Vasebi et al. 2013). Furthermore, different species of Trichoderma have been found to be effective in protecting the root system for some crops against other types or strains of pathogenic fungi, e.g., Fusarium solani (Mart.) Sacc. and M. phaseolina (Malik & Dawar 2003). Recently, Khalili et al. (2016) demonstrated that three T. harzianum isolates significantly inhibited the growth of M. phaseolina in vitro and field studies.

The aims of the present study were to evaluate the in vitro potential biological control of Trichoderma spp., Penicillium spp., and Aspergillus spp., against three species of Fusarium, and M. phaseolina, and to confirm their efficiency against the main cucurbit soil-borne pathogens in pot planted with melon and watermelon seedlings.

Material and Methods

Pathogen and antagonist strains

Twenty-two pathogens belonging to genus Fusarium, i.e., F. oxysporum f. sp. niveum, F. oxysporum f. sp. melonis, F. solani f. sp. cucurbitae, and M. phaseolina were used in vitro, and only eight were chosen for in vivo studies. Ten antagonist’s isolates were tested: four Trichoderma sp. (T. erinaceum Bissett, C.P. Kubicek & Szakács (watermelon); T. viride Schumach.; T. helicum Bissett, C.P. Kubicek & Szakács; and T. harzianum (melon)), two Penicillium sp. (P. digitatum (Pers.) Sacc. and P. italicum Wehmer (watermelon)), and four Aspergillus sp. (A. niger (melon), A. flavus Link, A. fumigatus Fresen., and A. terreus Thom (grafted watermelon)). The colonial and microscopic characteristics of the fungal isolates were determined. The pathogen and potential bioagents used in the present research were obtained from the Culture Collection Unit of the Laboratory of Phytopathology (ISA Chott Meriem, Sousse, Tunisia), and they were also isolated from infected cucurbit and tomato plants collected from agricultural fields in Tunisia (Table 1).
Table 1

Collection of 22 soil-borne pathogens isolates cucurbit plant host, regions, and date sampling. Four Fusarium oxysporum f. sp. niveum isolates, five Fusarium oxysporum f. sp. melonis isolates, eight Fusarium solani f. sp. cucurbitae isolates, and five Macrophomina phaseolina isolates

Pathogens

Code

Host

Regions

Sampling date

Fusarium oxysporum f. sp. niveum

FON1

Watermelon

Chebika

2009/2010

FON2

Jbeniana

2009/2010

FON3

Hajeb

2009/2010

FON4

Chott Meriem

2010

Fusarium oxysporum f. sp. melonis

FOM 1

Melon

Monastir

2011

FOM 3

 

2011

FOM 4

Kairouan sud

2011

FOM 6

 

2011

FOM 8

Sejnen

2011

Fusarium solani f. sp. cucurbitae

FSC1

Watermelon

Jbeniana

2010

FSC2

Hajeb

2010

FSC3

 

2010

FSC4

Beja

2010

FSC5

Chebika

2010

FSC6

Squash

Elkef

2010

FSC7

Watermelon

Jbeniana

2010

FSC8

Hajeb

2010

Macrophomina phaseolina

MP1

Melon

Chott Meriem

2011

MP2

Grafted watermelon

Chott Meriem

2011

MP3

Watermelon

Chott Meriem

2011

MP4

Tomato

Chott Meriem

2011

MP5

Melon

Chott Meriem

2011

In vitro experiment: antagonistic effect

Two disc plugs (0.5-cm diameter) of pathogen and antagonist (4 days old culture) were transferred respectively to a single potato dextrose agar (PDA) plate (9-cm diameter). The antagonist plug was placed on the one side of the plate (about 2 cm from the edge of the plate towards the center), while the pathogen plug was placed at the other side of the plate opposite to the antagonist plug leaving a distance of 5 cm between the two plugs. A plug of PDA medium was used as control treatment while the pathogen plug was placed at the other side. Three replications (two plates/replicate) for each individual treatment were made, and the plates were incubated at 28 ± 2 °C for 5 days. The inhibition percent of the radial growth was evaluated according to the formula of Hmouni et al. (1996): I (%) = (1 − C n /C0) × 100; where C n is the radial growth of the pathogen in the presence of the antagonist and C0 is the radial growth of control colonies.

In vivo experiment: evaluation of antagonist biological control activity

In vivo experiments were divided into two assays, the first one represented a preventive treatment of watermelon and melon against F. oxysporum f. sp. melonis and M. phaseolina root invasion, respectively. This assay was carried out by root-dipping watermelon and melon seedlings (15 days old) into flask containing a conidial suspension of the different antagonists for 30 min and 24 h before inoculation. For the curative assay, melon and watermelon seedlings were treated 24 h after inoculation with the pathogens by watering each plant with the antagonist suspension (10 ml) as mentioned in Table 2. Two cultivars of melon (cvs. Bonta and Anannas d’Amérique) and two of watermelon (cvs. Sirocco and Charleston Gray) were used in this assay. The seeds were sown in nursery seed trays with cells of volume 250 ml, with 15 plants per each treatment with 3 replicates (5 plants per replicate and treatment). The substrate used in the experiment consisted of a mixture of peat and vermiculite (1:1), which was autoclaved twice at 120 °C. The 2-l pots are then placed in a greenhouse for 30 days. Two positive controls were performed (one by inoculating the plants with only the pathogen and the other with the antagonist (10 ml)) and distilled water for the negative control. The experimental design was a randomized complete block design (RCBD), and the entire experiment was repeated twice. For each fungal species, one cucurbit plants randomly have been distributed in each treatment.
Table 2

Different treatments applied on watermelon and melon seedlings in vivo biological control assay. Two type of treatments: preventive (application of fungal antagonist before 24 h of pathogen) and curative (application of fungal antagonist 24 h after the inoculation)

Seedlings

Treatments

Preventive

Curative

Melon

Watermelon and melon

Melon

Watermelon

Pathogens

F. o. f. sp. melonis

M. phaseolina

F. s. f. sp. cucurbitae

F. o. f. sp. melonis

F. s. f. sp. cucurbitae

F. o. f. sp. niveum

Antagonists

FOM1/FOM6

MP1/MP2

FSC2/FSC5

FOM1/FOM6

FSC2/FSC5

FON1/FON2

Aspergillus flavus

+

+

+

+

+

+

A. fumigatus

+

+

A. niger

+

+

A. terreus

+

Penicillium italicum

+

+

P. digitatum

+

+

Trichoderma viride

+

+

+

+

+

+

T. harzianum

+

+

T. helicum

+

+

+

+

T. erinaceum

+

+

+

+

+ done, − not done

Inoculum preparation

For Fusarium species and bioagent fungi, the isolates were grown on PDA at 25 °C for 4 days until sporulation, and then, an Erlenmeyer flask containing 50 ml of potato dextrose broth (PDB, 20 g/l) was inoculated with four pieces individually. Spore production was induced in an orbital shaker, and the spores were recovered from culture by filtration. A hemocytometer was used to determine the concentration of the spores (106 spores/ml). In the case of M. phaseolina, the isolate were grown on PDA. Thus, 20 plates mixed with 2500 g of autoclaved potting mix and placed in 20-cm pots.

Evaluation parameters

At the end of the experiment, the plants were carefully removed from the pots, and the root systems were gently washed in tap water. Each root system was rated for the disease severity index (DSI) according to each pathogen. For F. oxysporum, we adopted the scale described by Vakalounakis and Frangkiadakis (1999) (0 = no symptoms; 1 = light vascular discoloration in the stem with or without stunting; 2 = vascular discoloration in the stem, stunting, wilting with or without yellowing of cotyledons; and 3 = dead seedlings). For F. solani f. sp. cucurbitae, the DSI was described by Boughalleb et al. (2005) (0: healthy; 1: slight yellowing of leaves with slight rot pivot and lateral roots and crown rot; 2: significant yellowing in leaves with or without wilting, stunting of plants, severe rot at the pivot and lateral roots, significant rot and browning of vessels in the stem; 3: death of the plant). For M. phaseolina, the scale used was described by Ravf and Ahmad (1998) (0: symptomless, 1: 1 to 3% of shoot tissues infected, 2: 10% of shoot tissues infected, 3: 25% of shoot tissues infected, 4: 50% of shoot tissues infected, and 5: more than 75% of shoot tissues infected). Other variables were measured to estimate the response of the cucurbits, such as the degree of inhibition exhibited by the antagonist: Damage reduction rate (R (%)) was calculated according to the two positive controls. Damage reduction of shoot and root dry weights: (R (%) = ((DWA − DWP) / DWA) × 100, which DWA is the dry weight (shoot and root) of inoculated plants with antagonist and DWP is the dry weight (shoot and root) of inoculated plant with only the pathogen.

The effect of the antagonist alone on the plants was also studied as the development rate of the dry shoot and root weights: D (%) = ((DWA − DW) / DW) × 100, where DWA is the dry weight (shoot and root) of the plants inoculated only by the antagonist and DW is the dry weight (shoot and root) of the healthy plants.

At the end of the curative treatment, agronomic parameters were determined including the shoot and root fresh (SFW and RFW) and dry weights (SDW and RDW) and the plant shoot and root height (SH and RH, respectively).

Statistical analysis

The data were analyzed by ANOVA using SPSS version 20.0 statistical software (SPSS, SAS Institute, USA) to evaluate parameter values differences. Differences between treatments were determined by Duncan’s multiple range test at 5% of significance level.

Results and discussion

In vitro experiment: antagonism effect

Data presented in Tables 3 and 4 indicated clearly that there was a significant reduction in mycelia growth after confrontation of tested pathogens with all antagonists. As shown in Table 3, the different species exhibited a significant reduction of mycelium growth of F. oxysporum f. sp. niveum which varied from 7.22 (FON2/Penicillium italicum) to 74.68% (FON4/Aspergillus flavus). This potential antagonist seemed to be the most effective bioagent with inhibition rate above 50%.
Table 3

Effect of direct dual confrontation, of two Penicillium spp. isolates, three Trichoderma spp. isolates, and Aspergillus flavus, on mycelia growth inhibition of four F. oxysporum f. sp. niveum isolates and eight F. solani f. sp. cucurbitae isolates after 5 days of incubation at 28 °C, means of six Petri plates (two plates per replicate)

Pathogens

Code

Mycelial growth inhibition percentage (%)a

 

P. digitatum

P. italicum

T. erinaceum

T. viride

T. helicum

A. flavus

P valuesb

F. oxysporum f. sp. niveum

FON1

40.56b AB

28.90c AB

46.08ab A

41.29b A

39.91b A

56.73a AB

> 0.05

FON2

32.10c BC

7.22d D

34.30b B

29.31c BC

31.00c AB

52.08a AB

> 0.05

FON3

32.98b BC

28.97b AB

40.92b AB

38.54b A

32.23b AB

64.71a A

0.0425

FON4

39.55b AB

30.44b A

42.66b A

36.95b AB

40.00b A

74.68a A

0.0335

F. solani f. sp. cucurbitae

FSC1

27.48bc C

21.89c C

37.08b AB

30.94bc B

29.23bc B

49.26a B

> 0.05

FSC2

38.02b B

24.92c BC

25.04c C

17.08c C

19.21c B

49.60a B

> 0.05

FSC3

44.65a A

26.19c B

21.83c CD

26.90c BC

17.44c C

48.94a B

> 0.05

FSC4

39.13b AB

28.57c AB

25.57c C

14.20c C

20.00c B

46.25a B

> 0.05

FSC5

28.35b C

25.21b B

26.13b C

31.00b B

24.91b B

48.89a B

> 0.05

FSC6

24.00b C

24.09b BC

19.60b CD

18.57b C

19.84b B

46.93a B

> 0.05

FSC7

35.85b B

24.27b BC

17.08bc D

10.93c D

19.28bc B

46.93a B

> 0.05

FSC8

19.49b D

17.74b CD

24.54b C

15.34b C

22.00b B

46.25a B

> 0.05

P valuesc

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

< 0.05

 

aMycelial growth inhibition percentage values; means of three replicates (two Petri plates per replicate). Duncan’s multiple range test: values followed by different letters are significantly different at P ≤ 0.05. Capital letters are for comparison of means in the same column. Small letters are for comparison of means in the same row

bDuncan’s multiple range test is for comparison of means among fungal antagonists with the same pathogen on mycelial growth inhibition

cDuncan’s multiple range test is for comparison of means among pathogens in the same fungal antagonist on mycelial growth inhibition

Table 4

Effect of direct confrontation of two Penicillium spp. isolates, two Trichoderma spp. isolates, and four Aspergillus sp. isolates on mycelial growth inhibition of F. oxysporum f. sp. melonis and M. phaseolina after 5 days of incubation at 28 °C, means of six Petri plates (two plates per replicate)

Pathogens

Code

Mycelial growth inhibition percentage (%)a

 

P. digitatum

P. italicum

T. viride

T. harzianum

A. flavus

A. niger

A. fumigatus

A. terreus

P valueb

F. oxysporum f. sp. melonis

FOM1

19.31b C

11.43c D

11.61c C

16.75b D

16.56b C

20.54b B

11.75b D

35.14a A

> 0.05

FOM3

20.38a B

19.73b B

10.24c D

19.8b B

25.95a AB

21.6a AB

26.35a A

18.81b C

> 0.05

FOM4

17.34b C

16.12b C

26.43a A

21.88a BC

22.1a BC

24.42a AB

21.26a BC

20.6a C

> 0.05

FOM6

19.91b B

23.27a AB

22.55a AB

16.6b C

25.71a AB

30.46a A

15.19b C

24.51a AB

> 0.05

FOM8

13.29c D

25.6b AB

8.71d D

23.31b C

36.59a A

27.86b AB

11.07c D

12.4c D

> 0.05

P valuec

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

 

M. phaseolina

MP1

48.01a A

36.22a A

42.43a A

42.38a A

23.77b B

29.33b B

35.97a A

21.84c B

> 0.05

MP2

38.54a B

35.39a B

38.15a B

52.42a A

33.51ab B

29.78a B

44.81a A

44.92a A

> 0.05

MP3

30.51ab A

46.63a A

37.90a B

46.21a A

42.56a A

30.63a B

24.54c C

24.64c C

> 0.05

MP4

27.09b C

43.97a A

33.27ab B

38.74b B

37.34a B

31.79a B

20.79c D

20.29cd D

> 0.05

MP5

35.14a B

31.95ab B

41.46a A

42.38a A

29.79b BC

27.29ab C

29.13bc BC

28.23b BC

> 0.05

P valuec

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

 

aMycelial growth inhibition percentage values; means of three replicates (two Petri plates per replicate). Duncan’s multiple range test: values followed by different letters are significantly different at P ≤ 0.05. Capital letters are for comparison of means in the same row. Small letters are for comparison of means in the same column

bDuncan’s multiple range test is for comparison of means among fungal antagonists with the same pathogen on mycelial growth inhibition

cDuncan’s multiple range test is for comparison of means among pathogens in the same fungal antagonist on mycelial growth inhibition

A. flavus reduced also the development of all F. solani f. sp. cucurbitae isolates and obtained data ranging from 46.25 (FSC8) to 49.6% (FSC2). However, F. solani f. sp. cucurbitae isolates showed a good resistance against the three Trichoderma species with inhibition rate below 30%. Among the different potential bioagents, Trichoderma erinaceum and Penicillium digitatum succeed to decrease the mycelial growth of F. oxysporum f. sp. niveum (41%) and F. solani f. sp. cucurbitae (32%) (Table 3).

The results from the dual culture tests are shown in Table 4. It appears that the growth rates of the five isolates of F. oxysporum f. sp. melonis differed according to the used antagonists. Three Aspergillus species exhibited growth inhibition and showed hyperparasitism against the colonies of F. oxysporum f. sp. melonis. Evaluation of the inhibition zones surrounding the A. flavus, A. niger, and A. terreus revealed an inhibition of 25%, with values comprised between 16.56 (FOM1/A. flavus) and 36.59% (FOM8/A. flavus) and from 20.54 (FOM1/A. niger) to 30.46% (FOM6/ A. niger) and 22% (from 18.81 (FOM3/A. terreus) to 35.14% (FOM1/A. terreus), respectively.

For M. phaseolina, the two Trichoderma species were revealed to be effective. In fact, the mycelial growth of the five M. phaseolina isolates decreased in the presence of Trichoderma harzianum with an average of 44.42% (values recorded between 38.74 and 52.42%) and Trichoderma viride (values ranged between 33.27 and 42.43%). P. italicum was the most efficient one with values between 31.95 (MP5) and 46.63% (MP3). However, values increased in the case of direct confrontation of M. phaseolina and the four Aspergillus species (Table 4).

In vitro biological control activity experiment revealed that A. flavus seemed to be the most effective bioagent with mycelial inhibition rate above 50% of F. oxysporum f. sp. niveum and it was able to reduce the mycelial growth of all F. solani f. sp. cucurbitae isolates, followed by T. erinaceum. The three Aspergillus species (A. flavus, A. niger, and A. terreus) and T. harzianum exhibited an important growth inhibition against the colonies of F. oxysporum f. sp. melonis. These results are in agreement with many reports. In fact, El-Sheshtawi et al. (2014) demonstrated that the presence of many biological control agents for Fusarium wilt which are able to exhibit high properties to inhibit conidial production over 90%, such as T. harzianum, Penicillium oxalicum Currie & Thom, and non-pathogenic F. oxysporum. Furthermore, Boughalleb et al. (2008) showed the good effect of three T. harzianum isolates against F. oxysporum f. sp. niveum and F. solani f. sp. cucurbitae, with a reduction of the colony diameter up to 50%. In the present research, among the tested potential bioagents, T. erinaceum and P. digitatum revealed to be able to decrease the mycelial growth of these two Fusarium species. In the same sense, Sreevidya and Gopalakrishnan (2016) found that the colony diameter of F. solani f. sp. cucurbitae was significantly decreased with Penicillium spp. used at higher concentration (75%). Dwivedi (2013) confirmed the fungi toxicity of four Aspergillus species (A. niger, A. flavus, A. sulphureus Desm., A. luchuensis Inui), two Trichoderma species (T. viride, T. koningii Oudem.), and two Penicillium species (P. citrinum Thom, P. italicum) against F. solani. This in vitro assay revealed that pathogenic fungi were significantly decreased even at low concentration of Aspergillus spp. Our findings for the two Trichoderma species against M. phaseolina were confirmed clearly. In fact, M. phaseolina mycelial growth decreased significantly in the presence of T. harzianum (44.42%). These results proved those of Khalili et al. (2016).

A microscopic study was conducted in order to determine the effects of some antagonists on the mycelial growth of F. oxysporum f. sp. melonis, F. oxysporum f. sp. niveum, and M. phaseolina (Fig. 1A, B). Compared to controls, treated Fusarium species mycelium showed strong lysis (Fig. 1B (a, f)), induction of mycelial cords via anastomosis between hyphal filaments (Fig. 1B (b)), mycelium winding (Fig. 1B (c, e)), and early chlamydospore formation (Fig. 1B (d, g)). The antagonistic effect is limited to not only the mycelial growth reduction but also the penetration, progression, colonization, and sporulation of the antagonist such as penetration sites of antagonist (Fig. 1B (i)) and lysis of M. phaseolina cells (Fig. 1B (j)).
Figure 1
Fig. 1

a Dual confrontation between two F. o. melonis FOM isolates and A. flavus, two F. o. niveum FON isolates and T. viride, and M. phaseolina MP3 and T. viride. b Mycelial interaction in vitro between F. o. melonis FOM (a, b, c, and d) and A. flavus, F. o. niveum FON (e, f, and g) and T. viride, and M. phaseolina MP3 (i and j) and T. viride after 5 days of incubation in PDA at 28 °C (Gr ×40) (one plate per treatment was illustrated). Three plates/treatment were taken for microscopic examination of F. o. melonis and F. o. niveum cultures, revealing lysis of fungal mycelia (a, f), mycelia cords (b), mycelium winding (c, e), and chlamydospore formation (d, g). Concerning M. phaseolina: i penetration sites of the antagonist and j lysis of the pathogen cells

In vitro antagonism by various antagonistic fungi on pathogenic organisms is a field of study in which reports are constantly thronging. High reduction of pathogen growth in vitro tests was observed by all antagonists. In this work, a microscopic observation of the interaction of F. oxysporum f. sp. melonis, F. oxysporum f. sp. niveum, and M. phaseolina and some antagonists confirmed the antibiosis such as penetration, progression, colonization, and sporulation. Similar results with other fungi have previously been reported by Benitez et al. (2004).

The possible mechanisms proposed to explain the antagonism were the competition. Therefore, one of the most interesting aspects of biological control is the study of the mechanisms employed by bioagents to reduce soil-borne disease incidence.

In vivo experiment: evaluation of antagonist biological control activity

The preventive and curative application of antagonist showed a good result with a disease incidence reduction in some agronomic traits in watermelon and melon reaching 50%.

Preventive treatment

Fusarium oxysporum f. sp. melonis

All tested antagonists for in vitro confrontation with F. oxysporum f. sp. melonis (FOM) were used for in vivo experiment. Both A. flavus and A. fumigatus have recorded the highest damage reduction rate of dry shoot and root weights of melon inoculated by FOM1 (41 and 52%) and FOM6 (42 and 53%). The in vivo effect of the biological agents were less noted for T. viride, and the values were ranged between 17.61 (FOM1) and 12.59% (FOM6), and 23.92% (FOM1 and FOM2), respectively. A. flavus and A. fumigatus significantly decreased the disease severity index (SDI) with values of 0.33 (FOM1) and 0.5 (FOM6) and with 0.5 (FOM1) and 0.67 (FOM6), respectively. The wilt was more apparent on inoculated melon plant treated with A. niger (1.67 (FOM1) and 1.83 (FOM6)) (Table 5). After 1 month of inoculation, the shoot and root dry weights of melon plants treated only with antagonists increased compared to non-treated plants (control). A. flavus induced the best results with an increase of shoot dry weights (11.45%) and of root dry weights (13%). T. harzianum (8.36 and 10.08%) and A. fumigatus (8.03 and 10.16%) were also effective in improving the plant development (Table 6).
Table 5

Damage reduction rate of shoot and root dry weight (%) and disease severity index values recorded by melon seedlings inoculated with two F. oxysporum f. sp. melonis isolates and treated preventively by four Aspergillus spp. isolates, two Penicillium spp. isolates, and two Trichoderma spp. in vivo assay

Antagonists

Damage reduction rate of shoot dry weight %a

Damage reduction rate of root dry weight %a

Disease severity indexa

FOM1

FOM6

FOM1

FOM6

FOM1

FOM6

A. flavus

40.61ab

42.15a

51.89a

53.2a

0.33b

0.5b

A. fumigatus

39.89a

42.25a

49.63a

52.55a

0.5b

0.67b

A. niger

21.73bc

18.28b

34.6b

21.27c

1.67a

1.83a

A. terreus

36.66ab

37.75a

46.86ab

42.19ab

0.83ab

0.67b

P. italicum

27.19abc

20.49b

38.04b

32.53abc

1ab

1b

P. digitatum

37.81ab

32.68a

47.17ab

32.53abc

0.83ab

0.67b

T. viride

17.61c

12.59b

23.92b

23.92c

1ab

1.33ab

T. harzianum

28.75abc

32.97a

40.87b

40.46ab

1ab

0.5b

P valueb

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

aDamage reduction rate values present the mean of three replicates (5 plants/replicate); R (%) = ((DWA − DWP) / DWA) × 100, where DWA is the dry weight (shoot and root) of inoculated plants with antagonist and DWP is the dry weight (shoot and root) of inoculated plant with only the pathogen and disease severity index scale (Table 3), which present the mean of three replicates (5 plants/replicate)

bDuncan’s multiple range test: values followed by different letters are significantly different at P ≤ 0.05

Table 6

Beneficial effect of four Aspergillus spp. isolates, two Penicillium spp. isolates, and two Trichoderma spp. isolates on melon plants in vivo assay, revealed by development rate of shoot and root dry weights (%)

Antagonists

Development rate of shoot dry weight %a

Development rate of root dry weight %a

A. flavus

11.45ab

12.87a

A. fumigatus

8.03abc

10.16ab

A. niger

1.72d

4.55b

A. terreus

3.24cd

7.5ab

P. italicum

4.13bcd

7ab

P. digitatum

5.41bcd

6.14ab

T. viride

7.03abc

9.26ab

T. harzianum

8.36ab

10.08ab

P value

> 0.05

> 0.05

aDevelopment rate presents the mean of three replicates (5 plants/replicate); D (%) = ((DWA − DW) / DW) × 100, where DWA is the dry weight (shoot and root) of the plants inoculated only by the antagonist and DW is the dry weight (shoot and root) of the healthy plants

bDuncan’s multiple range test: means followed by different letters are significantly different at P ≤ 0.05

M. phaseolina

The two Trichoderma species (T. harzianum and T. viride) and P. digitatum exhibited the highest damage reduction of shoot and root dry weights (R (%)) for inoculated watermelon plants. The damage reduction rate values were ranged between 51.11 and 40.42% (MP1) and from 52.71 to 33.56% (MP2) and varied from 54.33 to 27.78% (MP1) and between 52.71 and 33.57% (MP2), for the two parameters, respectively. However, the three Aspergillus species revealed to be less efficient. Watermelon plants treated with A. flavus, A. fumigatus, and A. niger showed symptoms on roots with highest disease severity index (2.33). However, the lowest value was exhibited on plant treated with T. viride (1.33). Both P. digitatum and T. viride recorded the highest values of damage reduction of shoot and root dry weights of melon plants. The lowest values of disease severity index was registered on plants treated with P. digitatum (0.83 for MP2) and with T. harzianum (0.55 for MP2) (Table 7). For watermelon plants, Trichoderma and Penicillium species exhibited the highest development rate (D (%)) ranging from 11.41 to 15.12% and from 7.76 to 15.17%, for the shoot and the root dry weights, respectively. However, the best behavior of melon plants was observed when they are treated with A. flavus, T. harzianum, and A. fumigatus (Table 8).
Table 7

Damage reduction rate of top and root dry weight (%) and disease severity index values recorded by watermelon and melon seedlings inoculated with two M. phaseolina isolates and treated preventively by four Aspergillus spp. isolates, two Penicillium spp. isolates, and two Trichoderma spp. isolates in vivo assay

 

Preventive treatment

Damage reduction rate of shoot dry weight %a

Damage reduction rate of root dry weight %a

Disease severity indexa

Cultivars

MP1

MP2

MP1

MP2

MP1

MP2

Watermelon

A. flavus

14.88bb

19.39c

11.73c

19.40c

2.33a

2.67a

A. fumigatus

8.94c

22.19b

35.09b

50.18a

2.33a

1.83abc

A. niger

31.97ab

27.70b

15.92c

27.70b

2.33a

1.50bc

P. italicum

13.75b

20.87b

26.85bc

43.00ab

2.17b

2.17ab

P. digitatum

43.36ab

33.56b

27.78bc

33.57abc

1.67b

1.50ab

T. viride

40.42ab

44.22b

34.30b

44.22ab

1.33b

1.33abc

T. harzianum

51.11a

52.71a

54.33a

52.71a

1.50b

1.50c

P valuec

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

Melon

A. flavus

3.69c

14.09c

18.31b

16.92d

1.33ab

2.00a

A. fumigatus

12.64c

28.40b

30.58ab

26.99cd

2.67a

1.33ab

A. niger

29.51ab

35.91b

26.29ab

37.35abc

1.50ab

1.33ab

P. italicum

7.38c

23.44c

20.25ab

27.90cd

1.50ab

1.50ab

P. digitatum

34.42a

39.51a

34.06a

33.30bcd

1.33ab

0.83b

T. viride

30.56ab

29.09b

35.41a

48.18ab

1.50ab

1.33ab

T. harzianum

27.80b

29.52b

37.23a

50.68a

1.33ab

0.50b

P valuec

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

aDamage reduction rate values present the mean of three replicates (5 plants/replicate); R (%) = ((DWA − DWP) / DWA) × 100, where DWA is the dry weight (shoot and root) of inoculated plants with antagonist and DWP is the dry weight (shoot and root) of inoculated plant with only the pathogen and disease severity index scale, which present the mean of three replicates (5 plants/replicate)

bDuncan’s multiple range test: means followed by different letters are significantly different at P ≤ 0.05

Table 8

Beneficial effect of four Aspergillus spp. isolates, two Penicillium spp. isolates, and two Trichoderma spp. isolates, on watermelon and melon plants in vivo assay, revealed by development rate of shoot and root dry weights (%)

 

Watermelon

Melon

Antagonists

Development rate of shoot dry weight %a

Development rate of root dry weight %a

Development rate of shoot dry weight %a

Development rate of root dry weight %a

A. flavus

5.98cb

4.99b

11.45a

12.87a

A. fumigatus

3.02c

3.44b

8.03ab

10.16ab

A. niger

3.57c

4.86b

1.72c

4.55b

P. italicum

11.41b

7.76ab

4.12bc

7.01ab

P. digitatum

13.93ab

14.64a

5.41bc

6.13ab

T. viride

11.91ab

10.32ab

7.03ab

9.26ab

T. harzianum

15.12a

15.17a

8.36ab

10.08ab

P valueb

> 0.05

> 0.05

> 0.05

> 0.05

aDevelopment rate presents the mean of three replicates (5 plants/replicate); D (%) = ((DWA − DW) / DW)× 100, where DWA is the dry weight (shoot and root) of the plants inoculated only by the antagonist and DW is the dry weight (shoot and root) of the healthy plants

bDuncan’s multiple range test: means followed by different letters are significantly different at P ≤ 0.05

Curative treatment

Watermelon

The efficiency of the three Trichoderma species and A. flavus, applied through plantation, on growth parameters was studied under greenhouse conditions. The results for F. solani f. sp. cucurbitae (FSC5) revealed that T. harzianum increased significantly the root height (16.6 cm), root fresh (0.47 g), and dry weight (0.23 g). In the same sense, the treatment with A. flavus produced the highest values of shoot height (49.6 cm) and dry weight (2.22 g). Watermelon plants inoculated with FSC2 and treated by T. viride exhibited a beneficial effect on shoot height (44.9 cm), fresh weight (6.14 g), and root dry weight (0.16). Plants inoculated by F. oxysporum f. sp. niveum and treated with the three Trichoderma species showed an improvement of the different growth parameters. The treatment with T. helicum generated the highest shoot height (TH) (61.3 cm) and increased also the shoot fresh weight (SFW) of watermelon plants inoculated with FON2 (12.6 g). T. viride and T. erinaceum improved the root fresh weight and the shoot and root dry weights with values of 0.75, 2.91, and 0.23 g, respectively (Table 9).
Table 9

Comparison of different growth parameter values: shoot and root heights (cm), shoot and root fresh weights (g), and shoot of root dry weights (g) recorded by watermelon seedlings inoculated by two F. solani f. sp. cucurbitae isolates (FSC 5 and FSC 2) and two F. oxysporum f. sp. niveum isolates (FON 1 and FON 2) and treated curatively by three Trichoderma spp. isolates and A. flavus

Pathogens

Treatments

Growth parameters

SH (cm)a

RH (cm)a

SFW (g)a

RFW (g)a

SDW (g)a

RDW (g)a

FSC5

T. erinaceum

46.80efghb

16.40abcde

6.46def

0.21ef

1.55defghi

0.15defg

T. viride

48.70defg

15.20cdefg

6.20efg

0.35cde

1.44fghij

0.17bcdef

T. helicum

44.00fghij

16.60abcd

5.21fgh

0.47bcd

1.63defgh

0.23ab

A. flavus

49.60def

16.40abcde

5.04fgh

0.27def

2.22b

0.18abcd

FSC5

40.80hij

13.50fgh

4.20hi

0.14f

1.90bcd

0.10g

FSC2

T. erinaceum

38.60ij

14.30defgh

5.61fgh

0.33cdef

1.47fghij

0.16cdef

T. viride

44.90efghi

16.90abc

6.14efg

0.31cdef

1.38ghij

0.16cdef

T. helicum

41.10hij

17.90ab

5.88fgh

0.28def

1.71cdefg

0.14defg

A. flavus

45.40efghi

18.60a

5.22fgh

0.29def

1.28hijk

0.15defg

FSC2

27.50k

12.20h

2.91i

0.18ef

0.97k

0.10g

FON1

T. erinaceum

51.70cde

15.80bcdef

8.21cd

0.49bc

2.91a

0.23a

T. viride

60.70ab

16.60abcd

10.77 b

0.75a

2.05bc

0.18bcde

T. helicum

61.30a

15.80bcdef

8.73c

0.33cdef

1.51efghi

0.13defg

A. flavus

57.00abc

14.00efgh

10.62 b

0.42cd

2.01bc

0.13defg

FON1

42.00ghij

12.10h

5.98fgh

0.18ef

1.12jk

0.14defg

FON2

T. erinaceum

54.20bcd

16.60abcd

11.73ab

0.71a

1.78cdef

0.18bcde

T. viride

50.30cdef

16.40abcde

11.21ab

0.65ab

1.46fghij

0.21abc

T. helicum

54.00bcd

16.00bcde

12.60a

0.51bc

1.86bcde

0.16cdef

A. flavus

46.10efgh

13.10gh

7.91cde

0.28def

1.79cdef

0.13defg

FON2

27.80k

12.40h

4.42ghi

0.20ef

0.99k

0.12fg

Control

 

37.00j

12.60h

5.08fgh

0.21ef

1.18ijk

0.13defg

P valueb

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

aShoot and root height (cm), shoot and root fresh weight (g), shoot of root dry weight (g), mean of three replicates (5 plants/replicate)

bDuncan’s multiple range test: means followed by different letters are significantly different at P ≤ 0.05

Melon

The best growth parameters on melon plants inoculated by FSC5 were recorded in the presence of T. erinaceum with values of 16.7 cm (RH), 7.31 (SFW), 0.46 g (RFW), 1.36 g (SDW), and 0.18 g (RDW), followed by T. viride treatments for plants inoculated by FSC2. In all the treatments for plants inoculated by F. oxysporum f. sp. melonis, significant reduction of disease incidence was noticed compared to control, especially for T. viride, T. erinaceum, and A. flavus (Table 10).
Table 10

Comparison of different growth parameters values: shoot and root heights (cm), shoot and root fresh weights (g), and shoot of root dry weights (g) recorded by melon seedlings inoculated by two F. solani f. sp. cucurbitae isolates (FSC 5 and FSC 2) and two F. oxysporum f. sp. melonis isolates (FOM) and treated curatively by three Trichoderma spp. isolates and A. flavus

Pathogens

Treatments

Growth parameters

TH (cm)a

RH (cm)a

TFW (g)a

RFW (g)a

TDW (g)a

RDW (g)a

FSC5

T. erinaceum

37.00abb

16.70a

7.31a

0.46a

1.36b

0.18a

T. viride

39.50a

14.90abc

6.99a

0.38ab

1.34b

0.12cdef

T. helicum

38.00a

11.20ef

5.14b

0.28bcde

1.00c

0.13cde

A. flavus

37.80a

14.10bcd

4.85b

0.27bcde

1.23b

0.17ab

FSC5

15.43g

11.00ef

3.78cde

0.21de

0.22g

0.08f0

FSC2

T. erinaceum

32.90c

16.70a

5.26b

0.34bc

1.39b

0.18a

T. viride

31.60c

15.50ab

7.36a

0.34bc

1.65a

0.15abc

T. helicum

30.90cd

14.80abc

5.23b

0.27bcde

1.36b

0.15abcd

A. flavus

33.70bc

16.60a

4.56bc

0.29bcd

1.28b

0.13bcd

FSC2

24.70ef

11.00ef

2.40f

0.17e

0.60de

0.09ef

FOM

T. erinaceum

22.50f

15.10ab

3.37def

0.23cde

0.51ef

0.09ef

T. viride

25.80ef

12.90cde

4.66bc

0.24cde

0.35fg

0.12cdef

T. helicum

25.00ef

13.60bcd

5.12b

0.26cde

0.53ef

0.13bcd

A. flavus

23.00f

15.30ab

4.35bcd

0.26bcde

0.29g

0.11def

FOM

15.30g

10.73f

3.28def

0.21de

0.23g

0.08f

Control

 

27.80de

12.50def

12.50def

0.22de

0.75d

0.17ab

P valueb

 

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

aShoot and root height (cm), shoot and root fresh weight (g), shoot of root dry weight (g), mean of three replicates (5 plants/replicate)

bDuncan’s multiple range test: means followed by different letters are significantly different at P ≤ 0.05

The present investigation revealed that the highest damage reduction rate of top and root dry weights was recorded on melon inoculated by F. oxysporum f. sp. melonis and treated preventively by A. flavus and A. fumigatus, also, decreased the disease severity index. A. flavus and T. harzianum were effective in improving the plant development. Among all curative treatments of inoculated melon plants by F. oxysporum f. sp. melonis, T. viride, T. erinaceum, and A. flavus were the most effective in reducing the disease incidence of this fungus. This stimulation results in greater axial growth and root mass compared to the control, which is consistent with the work of Mouria et al. (2007) who showed that all strains of T. harzianum stimulated the growth of tomato, including vegetative and root biomass. Several reports have previously demonstrated the successful use of biological control agents against Fusarium diseases of various crops. Bernal-Vicente et al. (2009) reported the specific biological control effect of T. harzianum against F. oxysporum f. sp. melonis under greenhouse nurseries. Watermelon and melon plants treated with P. digitatum and T. viride and inoculated with M. phaseolina recorded the highest damage reduction of shoot and root dry weights and the lowest disease severity index values. The development rate revealed the growth improvement induced by T. harzianum (watermelon, 15%) and A. flavus (melon, 12%). In fact, the application of Trichoderma to the soil as biological control agent in the greenhouse or under field conditions not only resulted in reduced disease severity of M. phaseolina but also enhanced plant growth (Srivastava et al. 2008). The efficacy of the three Trichoderma species and A. flavus, applied curatively of watermelon and melon, on growth parameters was studied under pot culture conditions. Watermelon plants inoculated with F. solani f. sp. cucurbitae and treated with T. erinaceum, T. viride, and A. flavus showed an improvement of growth parameters. T. helicum and A. flavus were effective on plants inoculated by F. oxysporum f. sp. niveum. The best growth parameters on melon plants inoculated by F. solani f. sp. cucurbitae were obtained in the case of T. erinaceum. Our results supported those of Harman et al. (2004) showing the use of Trichoderma spp. as plant growth enhancers, due to its production of growth hormones and enhanced transfer of minerals to the rhizosphere. The pathogen incidence and disease severity of plant inoculated only with pathogens were higher than the other treatments. Gava and Menezes (2012) revealed that Trichoderma spp. isolates have been shown to be efficient colonizers of the melon root system; however, the field efficacy did not exceed 50%. For M. phaseolina, watermelon and melon plants treated preventively with P. digitatum, T. harzianum, and T. viride recorded the highest values of damage reduction of shoot and root dry weights and the lowest disease severity index. Vasebi et al. (2013) determined the direct interaction between antagonist isolates and M. phaseolina involving increased fresh and dry weights of root and foliar parts, which supports my argument. Similar studies have previously shown that antagonists increase seed germination and promote plant growth (Sreedevi et al. 2011). Many studies demonstrated the promising results for Trichoderma species in the biological control of plant diseases applying the mechanisms of competition, antibiosis, and mycoparasitism mediated by hydrolytic enzymes (Munir et al. 2014). Trichoderma-based Trichoderma viride species have been investigated for over 80 years. Numerous researches have been focused on searching and selecting antagonist microorganisms on diverse soil pathogens. Also, synergism between different forms of action modes occurs as the natural condition for the biological control of fungal pathogens. It is widely known that environmental parameters such as abiotic (soil type, soil temperature, soil pH, water potential, and such like) and biotic (plant species and variety, microbial activity of the soil) factors as well as other factors such as method and timing of applications may have influence on the biological control efficacy.

Conclusions

Dipping watermelon and melon root in antagonists’ spore suspensions prior to inoculation of the culture substrate allowed not only the protection of the plants but also the improvement of the agronomic parameters, including better axial growth and greater root biomass. Aspergillus spp. were effective, applied preventively, in reducing F. oxysporum f. sp. melonis disease incidence. Furthermore, Trichoderma spp., applied preventively and curatively, showed a significant biological control activities on watermelon and melon plants inoculated with M. phaseolina, F. solani f. sp. cucurbitae, and F. oxysporum f. sp. niveum and could be recommended for biological control use. However, although Aspergillus spp. and Penicillium spp. were effective against the tested phytopathogens, fungi are not recommended for biological control assay due to their carcinogenic properties.

Declarations

Acknowledgements

This research was supported by UR13AGR03, University of Sousse, Tunisia. The experiments comply with the current laws of the country in which they were performed.

Authors’ contributions

All authors read and approved the final manuscript.

Competing interests

The authors declare that they have no competing interests.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors’ Affiliations

(1)
Département des Sciences Biologiques et de la Protection des Plantes, Université de Sousse, Institut Supérieur Agronomique de Chott Meriem Institut Superieur Agronomique de Chott Mariem, Sousse, Tunisia

References

  1. Benitez T, Rincon AM, Limon MC, Codon AC (2004) Biocontrol mechanism of Trichoderma strains. Int Microbiol 7:249–260PubMedGoogle Scholar
  2. Bernal-Vicente A, Ros M, Pascual JA (2009) Increased effectiveness of the Trichoderma harzianum isolate T-78 against Fusarium wilt on melon plants under nursery conditions. J Sci Food Agric 89:827–833View ArticleGoogle Scholar
  3. Boughalleb N, Armengol J, El Mahjoub M (2005) Detection of races 1 and 2 of Fusarium solani f. sp. cucurbitae and their distribution in watermelon fields in Tunisia. J Phytopathol 153:162–168View ArticleGoogle Scholar
  4. Boughalleb N, El Mahjoub M (2006) Fusarium solani f. sp. cucurbitae and F. oxysporum f. sp. niveum Inoculum densities in Tunisian soils and their effect on watermelon seedlings. Phytoparasitica 34(2):149–158View ArticleGoogle Scholar
  5. Boughalleb N, M’hamdi M, Romdhani MS (2008) Etude in vitro de l’activité antagoniste du Trichoderma harzianum vis-à-vis de Fusarium solani f. sp. cucurbitae et de Fusarium oxysporum f. sp. niveum. Revue de l’INAT 23(2):21–36Google Scholar
  6. De Cal A, Sztejnberg A, Sabuquillo P, Melgarejo P (2009) Management Fusarium wilt on melon and watermelon by Penicillium oxalicum. Biol Control 51:480–486View ArticleGoogle Scholar
  7. Dwivedi SKE (2013) In vitro efficacy of some fungal antagonists against Fusarium solani and Fusarium oxysporum f. sp. lycopersici causing brinjal and tomato wilt. Int J Biol Pharma Res 4(1):46–52Google Scholar
  8. El-Sheshtawi M, Bahkali AH, Al-Taisan WA, Elgorban AM (2014) Pathogenicity of Fusarium oxysporum f. sp. melonis to melon genotypes (Cucumis melo L.) and its biocontrol. J P A M 8(Spl. Edn. 1):317–324Google Scholar
  9. Eziashi EI, Omamor IB, Odigie EE (2007) Antagonism of Trichoderma viridae and effects of extracted water soluble compounds from Trichoderma species and benlate solution on Ceratocystis paradoxa. Afr J Biotechnol 6(4):388–392Google Scholar
  10. Fravel DR (2005) Commercialization and implementation of biocontrol (1). Annu Rev Phytopathol 43:337–359View ArticlePubMedGoogle Scholar
  11. Gava CAT, Menezes MEL (2012) Efficiency of Trichoderma spp. isolates on the control of soil-borne pathogens yellow melon in field conditions. Rev. Ciênc Agronômica 43:633–640View ArticleGoogle Scholar
  12. Hajieghrari B, Torabi-Giglou M, Mohammadi MR, Davari M (2008) Biological potential of some Iranian Trichoderma isolates in the control of soil borne plant pathogenic fungi. Afr J Biotechnol 7(8):967–972Google Scholar
  13. Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species—opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2:43–56View ArticlePubMedGoogle Scholar
  14. Hmouni A, Hajlaoui MR, Mlaiki A (1996) Résistance de Botrytis cinerea aux benzimidazoles et aux dicarboximides dans les cultures abritées de tomate en Tunisie. OEPP/EPPO Bull 26:697–705View ArticleGoogle Scholar
  15. Khalili E, Javed MA, Huyop F, Rayatpanah S, Jamshidi S, Abdul Wahab R (2016) Evaluation of Trichoderma isolates as potential biological control agent against soybean charcoal rot disease caused by Macrophomina phaseolina. Biotechnol & Biotechnological Equipment 30(3):479–488View ArticleGoogle Scholar
  16. Malik G, Dawar S (2003) Biological control of root infecting fungi with Trichoderma harzianum. Pak J Agric Sci 35:971–975Google Scholar
  17. Mouria B, Ouazzani-Touhami A, Douira A (2007) Effet de diverses souches de Trichoderma sur la croissance d’une culture de tomate en serre et leur aptitude à coloniser les racines et le substrat. Phytoprotection 88(3):103–110View ArticleGoogle Scholar
  18. Munir S, Jamal Q, Bano K, Sherwani SK, Abbas MN, Azam S, Kan A, Ali S, Anees M (2014) Trichoderma and biocontrol genes: review. Sci Agric 5:40–45Google Scholar
  19. Pavlou GC, Vakalounakis DJ (2005) Biological control of root and stem rot of greenhouse cucumber caused by Fusarium oxysponum f. sp. radicis-cucumerinum by lettuce soil amendments. Crop Prot 24:135–140View ArticleGoogle Scholar
  20. Ravf, B.A. and Ahmad, I. 1998. Studies on correlation of seed infection to field incidence of Alternaria alternate and Macrophomina phaseolina in sunflower. 13th Iranian Plant Protection Congress-Karaj. Iran, pp 113Google Scholar
  21. Shabir-U-Rehman WA, Ganie DSA, Bhat JA, Mir GH, Lawrence R, Narayan S, Singh PK (2013) Comparative efficacy of Trichoderma viride and Trichoderma harzianum against Fusarium oxysporum f. sp. ciceris causing wilt of chickpea. Afr J Microbiol Res 7(50):5731–5736View ArticleGoogle Scholar
  22. Sreedevi B, Charitha-Devi M, Saigopal DVR (2011) Isolation and screening of effective Trichoderma spp. against the root rot pathogen Macrophomina phaseolina. J Agric Technol 7(3):623–635Google Scholar
  23. Sreevidya M, Gopalakrishnan S (2016) Penicillium citrinum VFI-51 as bio agent to control charcoal rot of sorghum (Sorghum bicolor (L.) Moench). Afr J Microbiol Res 10(19):669–674View ArticleGoogle Scholar
  24. Srivastava JA, Singh RP, Srivastava AK, Saxena AK, Arora DK (2008) Growth promotion and charcoal rot management in chickpea by Trichoderma harzianum. J Plant Prot Res 48(1):81–92View ArticleGoogle Scholar
  25. Suárez-Estrella F, Vargas-García MC, López MJ, Capel C, Moreno J (2007) Antagonistic activity of bacteria and fungi from horticultural compost against Fusarium oxysporum f. sp. melonis. Crop Prot 26:46–53View ArticleGoogle Scholar
  26. Vakalounakis DJ, Fragkiadakis GA (1999) Genetic diversity of Fusarium oxysporum isolates from cucumber: differentiation by pathogenicity, vegetative compatibility, and RAPD fingerprinting. Phytopathology 89(2):161–168View ArticlePubMedGoogle Scholar
  27. Vasebi Y, Safaie N, Alizadeh A (2013) Biological control of soybean charcoal root rot disease using bacterial and fungal antagonists in vitro and greenhouse condition. J Crop Prot 2(2):139–150Google Scholar
  28. Woo SL, Ruocco M, Vinale F, Nigro M, Marra R, Lombardi N, Pascale A, Lanzuise S, Manganiello G, Lorito M (2014) Trichoderma-based products and their widespread use in agriculture. Open Mycol J 8:71–126View ArticleGoogle Scholar
  29. Zhang T, Shi ZQ, Hu LB, Cheng LG, Wang F (2008) Antifungal compounds from Bacillus subtilis B-FS06 inhibiting the growth of Aspergillus flavus. World J Microbiol Biotechnol 24:783–788View ArticleGoogle Scholar

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