African Journal of Biotechnology Vol. 7 (8), pp.

967-972, 17 April, 2008

Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2008 Academic Journals

Full Length Research Paper

Biological potantial of some Iranian Trichoderma

isolates in the control of soil borne plant pathogenic
fungi
Behzad Hajieghrari1*, Mousa Torabi-Giglou1, Mohammad Reza Mohammadi2 and Mahdi
Davari3.
1
Department of Plant Production, Moghan Junior College of Agriculture, University of Mohaghegh – Ardabili, Ardabil,
Iran.
2
Department of Plant Protection, Faculty of Agriculture, Islamic Azad University Branch, Varamin, Iran.
3
Department of Plant Protection, Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran.
Accepted 20 March, 2008

In this study the in vitro potential of six selected Iranian isolates of three species of Trichoderma
(Trichoderma hamatum T614, T. hamatum T612, Trichoderma harzianum T447, T. harzianum T969,
Trichoderma virens T523 and Trichoderma sp. T) were evaluated against five isolates of soil borne
phytopathogenic fungi (Fusarium graminearum, Rhizoctonia solani (AG4 and AG5), Macrophomina
phaseoli and Phytophtora cacturum) in dual culture techniques and through production of volatile and
non-volatile inhibitors, and the pH and temperature effects on Trichoderma mycelial growth were also
evaluated. All Trichoderma isolates had a marked statistical inhibitory effect on mycelial growth of the
pathogens in dual culture compared with controls. Maximum inhibitions occurred in F. graminearum-T.
hamatum T614 interaction. Significant pathogen colony growth inhibitions were observed when
exposed to the trapped atmosphere from culture of the Trichoderma. F. graminearum was most
susceptible to the volatile inhibitors produced by T. hamatum T612 (%inhibition = 48.65). Medium filtrate
obtained the Trichoderma isolate culture also were effected on the pathogen species significantly.
Maximum growth inhibition was observed in radial growth of F. graminearum by T. hamatum T612 non
volatile metabolites (%inhibition = 38.3). Evaluation of pH and temperature effects on Trichoderma
isolates mycelial growth showed that Trichoderma strains were found to be able to display activities
under a wider range of pH values. Also, Trichoderma strains are mesophilic.

Key words: Trichoderma spp., biological potantial, soil borne phytopathogenic fungi, Iran.

INTRODUCTION

Soil borne plant pathogens such as bacteria, fungi and new resistant strains of pathogens.
nematodes annually create a major economically losses In the recent years, there has been a world wide swing
in many important crops. Some chemical compounds to the use of eco-friendly methods for protecting the
have been successfully used to control soil borne plant crops from pest and disease (Rao et al., 1998). Biological
pathogens. Although in many cases, these pesticides control of plant disease especially soil borne plant
appear to be the most economical and efficient means of pathogens and nematodes by microorganisms has been
controlling plant pathogens. Toxicological environmental considered a more natural and environmentally accep-
and sociological concerns have led to drastic reduction in table alternative to the existing chemical treatment
the availability of efficient commercial compounds, and methods (Barker and Panlitz, 1996; Eziashi et al., 2007).
also the use of fungicides may lead to the appearance of Weindling (1932) over 75 years ago, demonstrated the
antagonistic nature of fungal species from the genus,
Trichoderma. The genus, Trichoderma is common fila-
mentous imperfect fungi (Deutromycetes, Dematiaceae),
*Corresponding author. E-mail: [email protected]. Tel: the most common saprophytic fungi in the rhizosphere
+989143186861.Fax: +984527463417. and found in almost any soil. The mycoparasite ability
968 Afr. J. Biotechnol.

of Trichoderma species against some economically im- area soil was evaluated. F. graminearum, R. solani AG4 and AG5,
portant aerial and soil borne plant pathogens (Papavizas, M. phaseoli and P. cactorum were isolated from infested wheat,
sugar beet, potato, soyabean and apple rootstock, respectively.
1985; Elad et al., 1993; Elad, 2000; Freeman et al., 2004, The isolates were maintained on potato dextrose agar (PDA)
Dubey et al., 2007) and nematodes (Windham et al., medium and stored at 4 C for further use.
1989; Sharon et al., 2001) allows for the development of
biocontrol strategies. Several Trichoderma species
reduces the incidence of soil borne plant pathogenic fungi Dual culture technique
under natural conditions (Sivan and Chet, 1986; Calvet et
al., 1990); however the efficacy of this depends largely on The Trichoderma isolates were evaluated against last mentioned
the physical, chemical and biological condition of soil. soil borne fungi by dual culture technique as described by Morton
and Strouble (1955). A 5 mm diameter mycelial disc from the
There have been numerous recent attempts to use margin of the Trichoderma 7 days-old culture of isolates and the
Trichoderma spp. on soil borne pathogens such as scle- soil borne pathogens were placed on the opposite of the plate at
rotinia, fusarium, pythium, and rhizoctonia species in equal distance from the periphery. The experimental design used
word (Elad et al., 1980; Jager et al., 1991; Ashrafizadeh was a completely randomized with four Petri dishes for each
et al., 2005; Dubey et al., 2007). Among these, several isolates. In control plates (without Trichoderma), a sterile agar disc
species of Trichoderma are well documented mycopara- was placed at opposite side of the soil borne inoculated isolates
plates. Inoculated plates were incubated at 25 ± 1oC until the end of
site and have been used successfully against certain the incubation period (7 days after inoculation). Two, 4, 6 days after
pathogenic fungi (Papavizas and Lumsden, 1980). T. the incubation period, radial growth of pathogen isolates was
harizanum, T. viridae, T. virens, T. hamatum, T. roseum measured and percent inhibition of average radial growth was
and T. koningii are the species that most often used calculated in relation to growth of the controls as follows:
biological control of pathogens. Trichoderma produced in
late years, have been developed into several commercial L = [(C – T)/C] x 100
biological control products to prevent development of
Where L is inhibition of radial mycelial growth; C is radial growth
several soil pathogenic fungi. The first biocontrol agent to measurement of the pathogen in control; T is radial growth of the
be commercialized registered and used in green house pathogen in the presence of Trichoderma isolates (Edington et al.,
crops and vineyards was isolate T39 of T. harzianum 1971).
®
(Trichodex by Machteshim, Israel) (Elad, 2000; Freeman
® ®
et al., 2004). ECOFIT (T. viridae) and TRI 002 (T.
harzianum) are marketed in India and Europe, respec- Slide culture method
tively, for control of various plant soil borne pathogens on
For each pathogen-Trichoderma interaction, a clean slide was
field and green house crops and vegetables (Koch, placed in 9 cm diameter plates and sterilized. Then a small amount
®
1999). SOILGARD (T. virens GL21) registered as of autoclaved melted potato dextrose agar was spread over the
biopesticide in the USA (Lumsden et al., 1996). Also slide to make a thin PDA film on the slide. The 5 mm discs of one
®
SUPERESIVIT (T. harzianum) has been reported to week old growing colonies cut from the margin of each pathogen
control Pythium ultimum (Duscova, 1995). In this regard, and Trichoderma isolates were placed on the opposite sides of the
the first requirement of biological control is the identi- slide 3 cm apart on the PDA surface. Then a few ml of double
distilled water was added to the plate to prevent drying and then
fication and deployment of highly effective strains control 25 ± 1oC for a week. At the end of incubation period,
of several soil borne plant pathogenic fungi in field crops meeting area of Trichoderma–Pathogen hyphae was observed
and green house system. under a light microscope for the presence of coiling structures for
In this study, the in vitro biological potential of some wall disintegration.
Iranian Trichoderma isolates (belonging to three species;
T. harzianum, T. hamatum, T. virens) were evaluated
against some common soil borne plant pathogens Effects of volatile metabolites
belonging to different group of fungi (F. graminearum,
The effect of the Trichoderma isolates of released volatile
Rhizoctonia solani AG4 and AG5, Macrophomina metabolites was evaluated on the mycelial growth of the pathogen
phaseoli and Phytophtora cactorum). Also, powerful and isolates by following the methods of Dennis and Webster (1971b).
highly effective isolates were selected for further field The 5 mm diameter mycelial disc of 7 days-old culture obtained
biocontrol studies. from the margin of each the Trichoderma isolates was centrally
placed on the PDA plates and incubated in 25 ± 1oC for 48 h. In
control plates, a 5 mm diameter of sterile PDA medium was placed
MATERIALS AND METHODS in the dish as done above. At the end of incubation period, the top
of each plate replaced with bottom of the PDA plate inoculated
Isolates centrally with 5 mm diameter mycelial plug of the pathogen isolates
and held together with adhesive tape. The experimental design
The Trichoderma isolates that were selected for this study were used was completely randomized with four replicates. Radial
obtained from collection of Trichoderma spp., in the Plant Pest and growths of the pathogens were recorded each day and
Disease Institute, Tehran, Iran. These include T. harzianum T447, percent inhibitions of average mycelial growth in relation to
T. harzianum T969, T. hamatum T612, T. hamatum T614 and T. growth of their controls were calculated by the last mentioned
virens T523. Also, one isolate of Trichoderma isolated from Moghan formula.
 Hajieghrari et al. 969

80
% Radial grow th inhibition 70
60 Rhizoctonia solani AG4
50 Macrophomina phaseoli
40 Phytophtora cactorum
30 Fusarium graminearum

20 Rhizoctonia solani AG5

10
0
14

12
23
T

69

47
.

T6

T6
T5
T9

T4
sp

ns
um

um
a

m
m

nu

nu
at

at
er

vi r
ia

ia
m

m
od

rz

rz
T.
ha

ha
i ch

ha

ha
T.

T.
Tr

T.

T.
Trichoderma isolates

Figure 1. Pathogen growth inhibition by Trichoderma isolates after 6 days of inoculation in dual culture.

Effects of non volatile metabolites RESULTS

To determine the effects of the Trichoderma isolate of non volatile In studying Trichoderma isolates and the pathogen spe-
metabolites on mycelia growth of the pathogens, three discs of cies in dual culture, all of the Trichoderma isolates had a
mycelial agar plugs (5 mm diameter) were removed with a No. 3 marked significant inhibitory effect on the growth of the
cork borer from the edge of the young culture. Trichoderma isolates
were inoculated in 100 ml sterilized potato dextrose broth (PDB) in pathogens compared with their control. Maximum patho-
250 ml conical flasks and incubated at 25 ± 1oC on a rotary shaker gens growth inhibitions occurred in interacting with T.
set at 100 rpm for 14 days. The control conical flasks were ino- harzianum T447. By 48 h after interaction between
culated with three 5 mm diameter of sterile PDA medium. The mycelia of Trichoderma isolates and the pathogens
culture was filtered through Millipore filter for removing mycelial mycelia, a clear zone of interaction was formed in all
mats and then sterilized through 0.2 µm pore biological membrane
Trichoderma-pathogen combination. In dual cultures of T.
filter (FP30/0.2 CA-S, Schleeicher and Schuell MicroScience
GmbH). virens T523 and T. harzianum T969 with all of the
The filtrate was added to molten PDA medium (at 40 ± 3oC) to pathogens, an inhibition zone without physical contact
obtain a final concentration of 10% (v/v). The medium was placed in between the colonies around the pathogen colony was
Petri dishes at 20 ml per plate and inoculated with 5 mm mycelial observed. And also, this inhibition zone without hyphae
plugs of the pathogens in the centre of the plates and incubated at contact was observed in R. solani AG4 and AG5-T.
25 ± 1oC for 7 days or until the colony reached the plate edge
(Dennis and Webster 1971c). There were 4 replicates for each
harzianum T447 and P. cactorum-Trichoderma sp. T inte-
treatment. Radial growths of the pathogens were recorded each raction. No apparent inhibition zone in other pathogen-
day. Percent inhibition of average growth mycelial in relation to Trichoderma interaction was observed macroscopically.
growth of the controls was calculated. Microscopically observation of the interaction zone
showed vacuolization and mycelial tip denaturizing of the
F. graminearum-Trichoderma virens T523 hyphae
pH and temperature followed by disintegration. No hyphae coiling was
observed in Pathogen-Trichoderma interactions. F.
To evaluate the influence of pH and temperature on the graminearum was most inhibited by T. hamatum T612
Trichoderma mycelial growth, a 5 mm diameter mycelial block cut
from the margin of 7-day old of each Trichoderma isolates colonies and R. solani AG4 was least inhibited by T. hamatum
by No. 3 cork borer was placed in PDA plates that adjusted pH to 5, T614 (Figure 1).
7, 8 with 0.1 N HCl and NaOH before autoclaving, and incubated at The significantly colony growth inhibition of the
20 ± 1oC, 25 ± 1oC and 30 ± 1oC The experiment was designed by pathogens was observed when exposed to the trapped
completely randomized design (Factorial). The colony diameters of atmosphere from cultures of the Trichoderma isolates in
the Trichoderma were measured in four replicates each day after
comparism with their control. F. graminearum was found
inoculation. The means were analyzed by analysis of variance
(ANOVA) and Least Significant Difference (LSD) test at 5% to be most susceptible to the volatile inhibitors produced
significant level with SAS software (SAS (1985) Institute Inc., Cary, by Trichoderma hamatum T612. The minimum inhibition
NC, USA). percentage was recorded in M. phaseoli-Trichoderma
970 Afr. J. Biotechnol.

60

50
%Radial growth inhibition

40 Rhizoctonia solani AG4

Macrophomina phaseoli
30 Phytophtora phaseoli
Fusarium graminearum
20 Rhizoctonia solani AG5

10

23
14

12
.T

69

47
T5
T6

T6
sp

T9

T4
ns
a

um

um
um

um
rm

ire
at

at
an

an
de

v
am

am
zi

zi
ho

T.
ar

ar
h

h
ic

h
T.

T.
Tr

T.

T.

Trichoderma isolates
Figure 2. Pathogen growth inhibition by Trichoderma volatile compounds.

45

40
%Radial growth inhibition

35

30 Rhizoctonia solani AG4

Macrophomina phaseoli
25
Phytophtora cactorum
20
Fusarium graminearum
15 Rhizoctonia solani AG5

10

0
14

12
23
T

69

47
.

T6

T6
T5
T9

T4
sp

um

um
ns
a

m
m

nu

nu
e
at

at
r

vir
de

zia

zia
m

m
ho

T.
ha

ha
r

r
ha

ha
ic

T.

T.
Tr

T.

T.

Trichoderma isolates

Figure 3. Pathogen growth inhibition by Trichoderma non-volatile compounds.

hamatum T614 interaction (Figure 2). Trichoderma isolates mycelial growth was statistically
Maximum growth inhibitions of F. graminearum radial differed in respect of the tested temperatures and pH.
o
growth were observed by T. hamatum T612 non-volatile 25 C supported the highest mycelial growth of T.
metabolites. Among M. phaseoli and R. solani, AG4 hamatum T612, T. harzianum T447, T. harzianum T969
showed minimum growth inhibition by non-volatile and T. hamatum T614. The mycelial growth of T. virens
o
inhibitors of T. hamatum T614 and Trichoderma sp. T, T523 and Trichoderma sp. T was highest at 30 C.
respectively. In addition, F. graminearum was more sus- Mycelial growth of T. hamatum T612, T. harzianum T447
ceptible to the inhibitory medium filtrate of all the tested and T. virens T523 was highest at pH 5 and mycelial
Trichoderma isolates (Figure 3). growth of T. harzianum T969 and Trichoderma sp. T was
 Hajieghrari et al. 971

Table 1. Effects of pH and temperature on the mycelial growth of Trichoderma isolates.

T. hamatum T. harzianum T. virens T. harzianum Trichoderma T. hamatum

Treatment T612 T447 T523 T969 sp. T T614
pH 8 33.26* 23.89 41.15 31.07 32.19 33.78
pH 7 29.3 24.3 33.96 39.7 39.45 30.85
pH 5 33.78 34.8 45.52 34.7 32.22 27.68
30°C 29.15 17.2 43.67 35.41 36.89 31.37
25°C 36.04 38.89 37.56 36.44 35.3 30.96
20°C 31.14 26.89 39.41 33.63 31.67 29.96

*Values are means of four replicates.

highest at pH 7. While T. hamatum T614 has the highest However, T. hamatum T612 is also a potential bioagent
mycelial growth at pH 8 (Table 1). for F. graminearum and a good candidate for further
study on F. graminearum biocontrol in field condition.
pH and temperature are two keys parameter to
DISSCUSION manipulate for growth, sporulation and saprophytic ability
as well as production of volatile and non-volatile
Plant pathogenic fungi and nematodes is a widespread metabolites, involved in nutrition, competition, mycopara-
problem and the use of chemicals is hardly successful. sitism, and extra cellular enzymes that disintegrate cell
However, the high cost associated with the use of wall of fungi. Therefore, it is important to collect
fungicides to control disease caused by soil borne fungi is information about the effects of pH and temperature on
a limiting factor in the profitability of crop production. the mycelial growth. It has been demonstrated that
According to this study, biological control could be the Trichoderma strains are active under a wider range of pH
best alternative and may be helpful, especially against (Kredics et al., 2003). The optimum temperature for
soil borne pathogens and nematodes. This is an integral growth differs among the Trichoderma isolates; although
part of the integrated pest management philosophy, most Trichoderma strains are mesophilic (Kredics et al.,
which entails the judicious use of biocontrol agents and 2003). The results obtained from present study also
reduced amounts of biocides/fungicides or other physical support these hypotheses.
aspects (such as soil solarization). The presence of an inhibition zone in dual culture
Trichoderma spp. that are common saprophytic fungi without the hyphae contact in treatments T. virens T523
found in almost any soil and rhizosphere micro flora, and T. harzianum T969 suggests the secretion of
have been investigated as potential biocontrol agents diffusible non-volatile inhibitory substance by the Tricho-
because of their ability to reduce the incidence of disease derma isolates. The results of non-volatile substance
caused by plant pathogenic fungi, particularly many against the pathogens that were more effective support
common soil borne pathogens (Papavizas, 1985; Sivan this hypothesis. However, it seems that 1:10 dilutions of
and Chet, 1986; Calvet et al., 1990; Elad et al., 1993; cultural filtrate used in this study were not in sufficiently
Elad et al., 1993; Spiegel and Chet, 1998; Elad, 2000; high concentrations to effect significant inhibitory
Freeman et al., 2004; Ashrafizadeh et al., 2005; Dubey et response. It is important to mention that Trichoderma
al., 2007), although some have been occasionally spp. are known to produce a number of antibiotics such
recorded as plant pathogens (Menzies, 1993). as Trichodernin, Trichodermol, Harzianum A and Harzi-
In this work, the results of dual culture revealed the anolide (Dennis and Webster, 1971c; Kucuk and
rapid colonization of the medium by Trichoderma Kivanc, 2004) as well as some cell walls degrading
isolates. All Trichoderma isolates evaluated were effec- enzymes such as chitinases, glucanases that break down
tive in controlling colony growth of the soil borne plant polysaccharides, chitins and -glucanase, thereby
pathogens. Evaluation of produced volatile and non- destroying cell wall integrity (Elad, 2000). These may also
volatile components also showed the acceptable perfor- play a major role in mycoparasitism because of changes
mance on inhibiting mycelial growth of pathogens. The in cell wall integrity prior than penetration.
results reported here suggest that from the six isolates of Selection of biocontrol agents as well as understanding
Trichoderma used in this study, T. harzianum T447, T. the mechanisms involved in the antagonistic effect of
hamatum T612 were more capable of influencing the Trichoderma spp. on plant pathogens are important in
growth of all tested pathogens in dual culture and through designing effective and safe biocontrol strategies.
production of volatile and non-volatile inhibitors under Different isolates of Trichoderma have various strategies
controlled condition, and may be used as a broad for fungal antagonism and indirect effects on plant health
spectrum biological control agents under field condition. also vary. Therefore, one of the most interesting aspects
972 Afr. J. Biotechnol.

of biology is the study of the mechanisms employed by Elad Y (2000). Biological control of foliar pathogens by means of
Trichoderma harzianum and potential modes of action. Crop Prot. 19:
biocontrol agents to affect disease control. Possible
709-714.
mechanisms of antagonism employed by Trichoderma Elad Y, Zimmand G, Zags Y, Zuriel S, Chet I (1993). Use of
spp. includes nutrient and niche competitions, antibiosis Trichoderma harzianum in combination or alternation with fungicides
by producing volatile components and non-volatile anti- to control Cucumber grey mold (botrytis cinerea) under commercial
greenhouse condition. Plant Pathol. 42: 324-356.
biotics (Harman and Hadar, 1983; Dennis and Webster,
Eziashi EI, Omamor IB, Odigie EE (2007). Antagonism of Trichoderma
1971b,c) that are inhibitory against a range of soil borne viridae and effects of extracted water soluble compounds from
fungi, as well as parasitism (Dennis and Webster, Trichoderma species and benlate solution on Ceratocystis paradoxa.
1971a). Also synergism between different forms of action Afr. J. Biotechnol. 6(4):388-392.
Freeman S, Minz D, Kolesnik I, Barbul O, Zreibil A, Maymon M, Nitzani
modes occurs as the natural condition for the biocontrol Y, Kirshner B, Rav-David D, Bilu A, Dag A, Shafir S, Elad Y (2004).
of fungal pathogens. Trichoderma biocontrol of Colletotrichum acutatum and Botrytis
It is widely known that environmental parameters such cinerea, and survival in strawberry. Eur. J. Plant Pathol. 110: 361-
as abiotic (soil type, soil temperature, soil pH, water 370.
Harman GE, Hadar Y (1983). Biological control of Pythium species.
potential and such like) and biotic (plant species and
Seed Sci. Technol. 11: 893-906.
variety, microbial activity of the soil) factors as well as Jager G, Velvis H, Lamers JG, Mulder A, Roosjen J (1991). Control of
other factors such as method and timing of applications Rhizoctonia solani in potato by biological, chemical and integrated
may have influence on the biological control efficacy of measures. Potato-Res. 34: 269-284.
Koch E (1999). Evaluation of commercial products for microbial control
Trichoderma isolates. Therefore, it is important that
of soil borne plant disease, Crop Prod. 18: 119-125.
Trichoderma biocontrol potential in field condition should Kredics L, Antal Z, Manczinger L, Szekeres A, Kevei F, Nagy E (2003).
be further evaluated. Influence of environmental parameters on Trichoderma strains with
biocontrol potential. Food Technol. Biotechnol. 41(1): 37-42.
Kucuk C, Kivanc M (2004). In vitro antifungal activity of strains of
Trichoderma harzianum. Turk. J. Biol. 28: 111-115.
ACKNOWLEGMENT Lumsden RD, Walter JF, Barker SR (1996). Development of
Gliocladium virens for damping off disease control. Can. J. Plant
We would like to express our special thanks to the Pathol. 8: 43-48.
Menzies JG (1993). A strain of Trichoderma viride pathogenic to
University of Mohaghegh Ardabili for its financial support germinating seedlings of cucumber, pepper and tomato. Plant Pathol.
of the research. 42: 784-791.
Morton DT, Stroube NH (1955). Antagonistic and stimulatory effects of
microorganism upon sclerotium rolfsii. Phytopathology, 45: 419-420.
REFERENCES Papavizas GC (1985). Trichoderma and Gliocladium biology, ecology
and the potential for biocontrol. Ann. Rev. Phytopathol. 23: 23-77.
Ashrafizadeh A, Etebarian HR, Zamanizadeh HR (2005). Evaluation of Papavizas GC, Lumsden RD (1980). Biological control soil borne fungal
Trichoderma isolates for biocontrolof Fusarium wilt of melon. Iranian propagules. Ann. Rev. Phytopathol. 18: 389-413.
J. Phytopathol. 41:39-57. Rao MS, Reddy PP, Nagesh M (1998). Evaluation of plant based
Barker R, Paulitz TC (1996). Theoretical basis for microbial interactions formulations on Trichoderma harzianum for the management of
leading to biological control of soil borne plant pathogens In: Hall R Meloidogyn incognita on egg plant. Nematol. Mediterr. 26: 59-62.
(Ed). Principals and practice of managing soilborne plant pathogens. Sharon E, Bar-Elad M, Chet I, Herrera-Estrella A, Kleifeld O, Spiegel Y
Am. Phythopathol. Soc. St. Paul, Mn. pp. 50-79. (2001). Biological control of root knot nematode Meloidogyne
Calvet C, Pera J, Bera JM (1990). Interaction of Trichoderma spp. with javanica by Trichoderma harzianum. Phytopathology, 91(7): 687-693.
Glomus mossaeae and two wilt pathogenic fungi. Agric. Ecosyst. Sivan A, Chet I (1986). Biological control of Fusarium spp. in cotton,
Environ. 9:59-65. Wheat and muskmelon by Trichoderma harzianum. J. Phytopathol.
Dennis C, Webster J (1971a). Antagonistic properties of species groups 116: 39-47.
of Trichoderma III, hyphae interaction. Trans. Br. Mycol. Soc. 57: Spiegel Y, Chet I (1998). Evaluation of Trichoderma spp. as biocontrol
363-369. agent against soil borne fungi and plant parasitic nematodes In Israel.
Dennis C, Webster J (1971b). Antagonistic properties of species groups Integr. Pest Manage. Rev. 3: 169-175.
of Trichoderma II, production of volatile antibiotics. Trans. Br. Mycol. Statistical Analysis Software (SAS) (1985). Users Guide: Statistics
Soc. 57: 41-47. version 5 Edition. SAS Institute Inc. Cary. NC. p. 956.
Dennis C, Webster J (1971c). Antagonistic properties of species groups Weindling R (1932). Trichoderma lignorum as a parasite of other soil
of Trichoderma I, production of non-volatile antibiotics. Trans. Br. fungi. Phytopathology, 22: 837.
Mycol. Soc. 57: 25-39. Windham GL, Windham MT, Williams WP (1989). Effects of
Dubey SC, Suresh M, Singh B (2007). Evaluationof Trichoderma Trichoderma spp on Maize growth and Meloidogyne arenaria
species against Fusarium oxysporum fsp. Ciceris for integrated reproduction. Plant Dis. 73: 493-494.
management of chickpea wilt. Biol. Contr. 40: 118-127.
Duscova E (1995). New biological fungicides for plant protection
registered in the Czech Republic In: Manaka M (Ed). Environmental
biotic factors in integrated plant disease control, Poznan. 5-9
September 1994. Polish Phytopathol. Soc., Poznan. pp. 211-216.
Edington LV, Khew KL, Barron GI (1971). Fungitoxic spectrum of
benzimidazole compounds. Phytopathology, 61:42-44.
Elad T, Chet J, Katan J (1980). Trichoderma harzianum a biocontrol
effective against Sclerotium rolfsii and Rhizoctonia solani.
Phytopathology, 70: 119-121.