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R Renuka1, B. Parameswari2 and M.L. Chhabra3


Assistant Professor, Pandit Jawaharlal Nehru College of Agriculture &

Research Institute, Puducherry.


Scientist, Sugarcane Breeding Institute Regional Centre, Karnal.

Senior Scientist, Sugarcane Breeding Institute Regional Centre, Karnal.

Groundnut (Arachis hypogea L.), which occupies a prime position among the
commercially cultivated oil seed crops in India. It is an important food source with a high










per cent edible oil. A large number of diseases attack groundnut in India and the majority are
caused by fungi and several of them are yield reducers in certain regions. Among the fungal
diseases, root rot caused by Macrophominha pahseolina (Tassi) Goid., a warm dry weather
pathogen, is responsible for economic losses to an extent of 80 per cent. The fungus is seed as
well as soilborne pathogen and propagules of the pathogen are distributed randomly in soil
and often cannot be controlled by fungicides. Moreover, the fungicides are effective only on
the active metabolic stage of the propagules and not on resting structures. However, excessive
use of chemical fungicides in agriculture has led to deteriorating human health,
environmental pollution, and development of pathogen resistance to fungicide. Because of
the worsening problems in fungal disease control, a serious search is needed to identify
alternative methods for plant protection, which are less dependent on chemicals and are more
environmentally friendly.
Biological control has become a critical component of plant disease management and
it is a practical and safe approach in various crops. Among the bio agents, utilization of the
plants own defense mechanism induced by endophytes is the subject of current interest in the
management of pests and diseases. Biological control with endophytes offers an effective
strategy for disease management. Recently, researchers have reported that several fungal and
bacterial endophytes promote plant growth, help to withstand against pathogen attack by
competition, antibiosis and eliciting Induced Systemic Resistance (ISR). Exploiting an
additional microbial habitat for bio control purposes might enhance overall bio-control
efficacy and increase consistency in performance, since the endophytic agent could avoid
unfavorable conditions prevailing in the environment by entering and localizing in the
intercellular spaces of the epidermal cells of plant tissues. In biocontrol methods, the use of
*Corresponding authors e-mail: pathorenu@yahoo.co.in

endophytic bacteria against many plant pathogens has been well documented by several
authors. Endophytic bacterial isolate EPC5 (Plant growth promoting endophytic bacteria),
from coconut have been shown to effectively inhibit the Ganoderma lucidum growth in vitro.
Endophytic bacillus amended with chitin promotes higher growth and suppresses bacterial
blight incidence in cotton under greenhouse conditions.
Groundnut or peanut (Arachis hypogaea L.) an important annual oil seed crop and
protein source of the smallholder-farming sector in India, is susceptible to root rot caused by
M. pheasolina, affecting the quality and quantity of crop, causing severe damage with yield
reduction. Biocontrol of soil borne phytopathogens by antagonistic bacteria offers a non
polluting complement and an alternative to existing disease management strategies. The
major factor responsible for the inconsistency of the introduced biocontrol agent is its
variable colonization ability, which is influenced by various biotic and abiotic factors. As
endophytes are sheltered by the host plant from environmental stresses and microbial
competition and colonize the same niche as plant pathogens, they may be better suited for
biological control by either outcompeting or directly antagonizing pathogens. Though the
efficacy of several biocontrol agents like Pseudomonas fluorescens, Bacillus subtilis and T.
viride has been reported against M. phaseolina, role of endophytic bacteria against the root
rot of groundnut have not been studied in depth. Hence, the present study was undertaken to
investigate the effect of groundnut associated endophytic bacteria against root rot pathogen
M. phaseolina.
Isolation of endophytic bacteria
During the research work totally fourty five isolates of endophytic bacteria were
isolated from healthy leaf, stem and root of groundnut plants collected from six districts of
Tamil Nadu. Totally nineteen bacterial isolates were obtained from leaves, fourteen from stem
and twelve from roots of healthy groundnut plants. More number of isolates (13 isolates)
were obtained from Pudukottai district followed by Kallakurichi and Salem (each 8 isoltes).
The isolates were tentatively named as in the table 2 and maintained on Nutrient agar slants
for further screening. Similarly fifty five isolates of endophytic bacteria were isolated from
healthy coconut roots from different parts of Tamil Nadu by Rajendran et al., (2007). Hirano
and Upper, (2000) reported that endophytic population is dominated by bacteria which can be
found in numbers ranging 105 to 107 cells per gram of plant material. Melnick
et al., (2011) isolated sixty-nine endospore-forming bacterial endophytes consisting of 15
different species of five genera from leaves, pods, branches, and flower cushions of
Theobroma cacao as potential biological control agents.

*Corresponding authors e-mail: pathorenu@yahoo.co.in

In vitro screening of the endophytic bacterial strains against the pathogen

Those isolates were tested for their efficacy against M. phaseolina by dual plate
technique. Among the fourty five isolates, thirty five were found to inhibit the growth of
M. phaseolina under in vitro condition and others showed a minimum zone of inhibition, as
the fungus stopped growing near the bacterial streak and in six of the bacterial isolates the
pathogen overgrew it. Six of these isolates showed the maximum antagonism against
M. phaseolina. The isolate EPG18 (leaf isolate from Tindivanam) caused highest inhibition of
M. phaseolina followed by EPG10 (leaf isolate from Pudukottai) and EPG7 (leaf isolate from
Dharmapuri). The per cent inhibition was significantly higher in plates streaked with EPG18
(62.57 per cent) followed by EPG10 (60. 21 per cent) which is on par with EPG7 (60.09 per
cent) against control plates These three isolates effectively inhibited the growth of pathogen
even up to one month whereas in control, M. phaseolina overgrew the plate within 6 days
after placing the disc. The antagonistic activity of endophytes against the soil the borne
pathogen G. lucidum is also reported by Rajendran et al., (2007). The endophytic bacteria
from the coconut palms EPC5, EPC8, EPC15, EPC29 and EPC52 inhibited the growth of G.

under in vitro and were found to increase the vigour index of rice seedlings

significantly when compared to untreated control. The result was in conformity with Hui Li
et al., (2012) who reported that the culture filtrate and the n-butanol extract of an endophytic
bacterial Bacillus subtilis isolated from healthy stems of Prunus mume showed strong growth
inhibition activity in vitro against the replant disease phytopathogens Fusarium
graminearum, Alternaria alternata, Rhizoctonia solani, Cryphonectria parasitica and
Glomerella glycines.
Endophyte treatment played a dual role by promoting plant growth and reducing
disease severity and it was comparable with chemical treatment. Many are capable of
synthesizing bioactive compounds that can be used by the plant for defense against
pathogenic microorganisms (Schulz et al., 2002; Strobel, 2001; Corrado & Rodrigues, 2004;
Owen & Hundley, 2004; Gimnez et al., 2007). The endophytic Bacillus spp. CY22 isolated
from balloon flower produced iturin A with antifungal activity against Rhizoctonia solani,
Pythium ultimum and Fusarium oxysporum (Cho et al. 2003). Hallman et al. (1997) reported
that most of the endophytic bacterial strains are capable of promoting plant growth.
Pseudomonas fluorescens strain G8-4, which was later designated 89B-61 and found to
colonize plants internally, elicited systemic protection against cucumber anthracnose
following application to cucumber seeds. Of all the tested bacterial endophytes, leaf isolate

*Corresponding authors e-mail: pathorenu@yahoo.co.in

EPG18, showed the most effective inhibition against M. phaseolina followed by the isolates
EPG10 and EPG7.

*Corresponding authors e-mail: pathorenu@yahoo.co.in