Professional Documents
Culture Documents
ABSTRACT
Introduction
Trichoderma is a genus of fungi that is characteristic sweet or coconut odour
present in all soils, where they are the (Aneja, 2003). Trichoderma harzianum is
most prevalent culturable fungi. Many a common soil, litter, and wood fungus. It
species in this genus can be characterized possesses highly cellulolytic activity and is
as opportunistic avirulent plant symbionts. main agents of decomposition. Several
Cultures are typically fast growing at 25- strains of Trichoderma have been
30°C, but will not grow at 35°C. Colonies developed as biocontrol agents against
are transparent at first on media such as fungal diseases of plants. The various
cornmeal dextrose agar (CMD) or white mechanisms include antibiosis, parasitism,
on richer media such as potato dextrose inducing host-plant resistance, and
agar (PDA). Some species produce a competition. Most biocontrol agents are
96
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
from the species T. harzianum, T. viride seedling emergence was observed and
and T. hamatum. Trichoderma species are these plants exhibited a 95% increase in
an efficient biocontrol agent that is root area. Similarly an increase in
commercially produced to prevent Phosphorous and Iron concentration was
development of several soil pathogenic observed in Trichoderma inoculated
fungi. Different mechanisms have been plants. The Trichoderma species release
suggested as being responsible for their antibiotics and other chemicals that are
biocontrol activity, which include harmful to pathogens and inhibit growth
competition for space and nutrients, (antibiosis). The potential use of the
secretion of chitinolytic enzymes, Trichoderma species as a biocontrol agent
mycoparasitism and production of was suggested more than 70 years ago by
inhibitory compound. Most plant species Weindling (1932) who was first to
exploit soil with the help of beneficial demonstrate the parasitic activity of a
microorganisms such as fungi and member of this genus against soilborne
bacteria, some of which are important in fungal or bacterial pathogens. The
nitrogen fixation and phosphate mechanisms proposed to explain the
solubilization. Occurrence and distribution biocontrol of plant pathogens by
of phosphate solubilizing microorganisms Trichoderma are presumptive.The
(PSM) have been found in almost all the suggested mechanisms for biocontrol are
soils tested, although their populations antibiosis, lysis, competition, and
vary with different soils, climate, and mycoparasitism (Cook and Baker 1983).
cropping history (Kucey, 1989). These may act alone or in combination.
Trichoderma species are also effective
Many recent findings suggest that plant against various Gram positive and Gram
development and biochemistry are negative bacterial species. They produce
strongly affected by Trichoderma strains among 40 different metabolites of
(Mach et al., 1993). Nutrient competition, Trichoderma harzianum and ciprofloxacin
mechanical barriers, or pH changes are and norfloxacin in cultures of
some of the antagonistic effects. Both Trichoderma viridae which are
fungi and bacteria are able to synthesize a antibacterial in nature.
wide range of metabolites with fungicidal
and bactericidal capabilities. These Materials and Methods
antibiotics are an alternative biological
protection to conventional fungicides Isolation of Microorganisms
(Brown 1996). Specific strains of fungi in
the genus Trichoderma colonize and To isolate Trichoderma from soil, a
penetrate plant root tissues and initiate a Trichoderma selective medium was
series of morphological and biochemical prepared (Mohammad Akrami et al.
changes in the plant, considered to be part 2011). The basal medium consisted of 0.2
of the plant defense response, which in the g MgSO4 (7H2O), 0.9 g K2HPO4, 0.15 g
end leads to induced systemic resistance KCl, 1.0 g NH4NO3, 3.0 g D+ glucose
(ISR) in the entire plant. The capability anhydrous, 0.15 g rose bengal and 20g
of Trichoderma species to promote agar. These constituents were added to 950
increased growth response was verified ml of distilled water and autoclaved at
both in greenhouse experiments and in the 121°C for 30 min. The biocidal
hydroponic system. A 30% increase in ingredients, 0.25g chloramphenicol, is
97
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
98
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
organisms were performed using SDA (Fethi Bel Haj 2008, Parshikov et al.
medium, control tubes were also 2002). The concentrations at which the
maintained. The tubes were kept in bacterial species Staphylococcus aureus,
slanting position for 30 minutes to obtain E coli, Klebsiella were found to be
SDA slants. The slants were seeded with inhibited by the Trichoderma harzianum
test organisms. The inoculated slants were extract were 100 µl/ml of T. harzianum
incubated at 37° C for 24 hours for extract A niger & A clavatus among fungal
bacteria and for fungi 25 - 28 °C for 72 isolates and Proteus among bacteria were
hours. The concentration of the resistant to antimicrobial activity of T
Trichoderma harzianum extract at which harzianum extract.
there is no growth of organism was
recorded and considered as MIC of the Trichoderma produce or release a variety
extract of that organism. The results were of compounds that induce localized or
expressed in terms of growth of organisms systemic resistance responses, and this
(+) or inhibition of growth (-). The lowest explains their lack of pathogenicity to
concentration inhibiting fungal growth plants. These root-microorganism
was noted as MIC (Rath et al., 1999). associations cause substantial changes to
the plant proteome and metabolism. Plants
Results and Discussion are protected from numerous classes of
plant pathogen by responses that are
The concentrations at which the fungal similar to systemic acquired resistance and
species Aspergillus fumigatus, Aspergillus rhizobacteria-induced systemic resistance.
flavus, Aspergillus candidus, Pesticide and organic compound that are
Cladosporium, Rhizopus were found to be widely used to control plant pathogen do
inhibited was 100µl/ml, whereas for not degrade completely and leave toxic
Aspergillus niger, Fusarium graminarium, residue. Hence scientist has turned on to
Fusarium semitectum, Aspergillus terreus fungus to their use as a controlling agent.
were found to inhibit by the Trichoderma Several various experiment have already
harzianum extract at 150 µl/ml (Table 1 been reported showing fungal inhibiting
and 2). various other plant pathogen.
Trichoderma harzianum isolate showed
Based on the biochemical tests the strong antagonism against fungal species
Bacteria were E.coli, Klebsiella, (Aspergillus niger, Aspergillus fumigates,
Staphylococcus aureus, Proteus. The fungi Aspergillus flavus, Aspergillus terreus,
include A. flavus, A. clavatus, A. terreus, Aspergillus candidus, Fusarium
A. fumigatus, A. candidus, A. niger, graminarium, Fusarium semitectum,
Fusarium graminiarum, F. semitechtum, Cladosporium, Rhizopus). The same
Rhizopus, Cladosporium. In our studies phenomenon was observed by Nashwa
the strain of Trichoderma also showed a M.A. Sallam et al., (2008); Jegathambigai
various degree of inhibition to various et al., (2009). The main theme of our
plant pathogens (Table 3 and 4). When experiment is to show at what
Trichoderma harzianum tested for their concentration does the fungus inhibits or
antagonistic activity against the bacterial shows the maximum inhibition of growth
species (Staphylococcus aureus, Proteus, of other pathogen. When Trichoderma
E. coli, Klebsiella) the isolates were found harzianum was inoculated against
to be affective at various concentration different plant pathogens that are
99
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
100
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
5 10 25 50 100
1. Staphylococcus aureus 1.0 1.0 1.2 1.5 2.0
2. Proteus 0.8 1.0 1.2 1.2 1.8
3. E. coli 1.0 1.2 1.4 1.6 1.9
4. Klebsiella 0.9 1.2 1.5 1.7 1.9
been isolated from different source of fertilizers to cut down their expense on
infected plants by standard blot method. chemical fertilizers.
The inoculated organism showed a varying
degree of inhibition. References
101
Int.J.Curr.Microbiol.App.Sci (2014) 3(1): 96-103
103