Trends in Biosciences 9(10), Print : ISSN 0974-8431, 623-626, 2016
Biotoxicity of Some Indigenously Available Plant Leaf Extracts Against
Root-Rot Fungus Rhizoctonia solani Kuhn
MUCKSOOD AHMAD GANAIE* AND TABREIZ AHMAD KHAN
Section of Plant Pathology and Nematology, Department of Botany,
Aligarh Muslim University, Aligarh, India.
ABSTRACT
Control of root rot fungus generally involves the use
of synthetic fungicides. However, apart from it’s high
cost, increased concentration for the environment has
necessitated a reduction in the amount of fungicides.
In the present study leaf extracts of some indigenously
available plants leaves were assessed to determine
their efficacy against mycelial growth and scleriotia
formation of root-rot fungus Rhizoctonia solani. The
leaf extracts varied in their antifungal properties
against the R. solani. Some inhibited the growth where
as others had less or no effect. The maximum
inhibitory effect on mycelial growth was shown by
leaf extract of Ipomea palmata followed by Bauhinia
variegate, Quadqualis indica, Hemelia patens ,
Bougainvillia spectabilis , and Polyalthia longifolia.
However, the leaf extract of Nerium indicum and
Callistemone lanceolata were found to show less than
20% growth inhibition of test pathogen as compared
to control. No inhibitory effect was shown by the leaf
extracts of Clerodendron splendensens, and Thevetia
peruviana. Similarly, the greatest inhibition in the
development of scleriotia was found by the leaf extract
of Bauhinia variegata followed by Ipomea palmata,
Quadqualis indica, Hemelia patens, Bougainvillia
spectabilis, Polyalthia longifolia , Callistemone
lanceolata, Nerium indicum. Moreover the least
production of sclerotia was recorded by Thevetia
peruviana and Clerodendron splendensens leaf
extracts.
Keywords Leaf extracts, R. solani, Mycelial
growth, Sclerotia formation
Fungi are extremely diverse and common plant
pathogens. All the plants are attacked by some kind
of fungi and each of the parasitic fungi can attack
one or many kinds of plants. More than 70% of all
major crop diseases are caused by fungi and led to
significant yield losses in most of the agricultural
and horticultural crops (Agrios, 2005). Most of the
important fungal pathogens involved in the
production of diseases in plants including
ornamentals are soil borne root infecting fungi viz.,
Macrophomina phaseolina (Tassi) Goid.,
Rhizoctonia solani Kuhn., Fusarium spp., Pythium
spp., Phytophthora spp., Sclerotium spp. and
Verticillium spp. Among these fungi, Rhizoctonia
diseases occur worldwide on almost all vegetables,
multiple field crops, turf grasses, shrubs and trees.
Synthetic pesticides are nowadays widely
used for the control of plant diseases throughout
the world because of their higher effectiveness in
controlling disease causing organisms. However,
indiscriminate use of synthetic chemicals for
controlling plant diseases is likely to give rise to
phytotoxicity, environmental pollution and
resistance to plants against the pathogens.
Therefore, there is an urgent need to search for
effective, safe and biodegradable alternative
pesticides. In many countries, efforts are being
made to minimize the use of harmful insecticides
through the use of indigenous plant products,
implementation of IPM approaches, use of bio-
degradable products (Khattach and Hameed, 1986).
Botanicals are materials/products made or
derived from plants. The botanicals used for
protection against pest can be termed as ‘Botanical
pesticides’. These botanicals rapidly degrade in the
environment and are of less threat to other
organisms. These are also easily available and cheap
in tropical and sub tropical countries. Botanical
pesticides are natural plant products that belong to
the so called secondary metabolites, which include
alkaloids, terpenoids, phenolics, and minor
secondary chemicals. Plants are rich source of
bioactive organic chemicals. It is estimated that
the plants may contain as many as 4000,000
secondary metabolites (Mamun,2011).
The pest control properties of plants can be
utilized either directly by using plant tissues and/or
dry powder, or crude derivative, such as organic
extract, or if possible through industrial process
after isolation and identification of the active
compound/ compounds of plants (Grainge and
Ahmad, 1988). The presence of antifungal
624 Trends in Biosciences 9 (10), 2016

Table 1. Effect of leaf extract of some plants on the mycelial growth and sclerotial formation of
Rhizoctonia solani.

Name of plant Diameter of mycelial mat after Per cent No. of Per cent
decrease sclerotia decrease
24 48 72 96 120 144 168
or per or
hr hr hr hr hr hr hr
increase petridish increase
after 168
hrs
Clariodendron 0.93 1.33 3.0 4.25 5.6 8.86 10.0 0 125.45 9
splendensens
Nerium indicum 0.40 1.13 2.16 3.3 4.30 5.76 8.66 13 98.7 28
Thevetia 0.80 1.66 2.66 3.93 4.86 8.73 10.0 0 130.2 6
peruviana
Bauhinia variegata 0.70 1.26 2.55 3.80 4.50 5.20 5.65 43 62.45 55
Polyalthia 0.30 1.0 2.5 2.76 3.73 5.10 7.53 24 89.5 35
longifolia
Hemelia patens 0.93 1.60 3.30 3.76 4.20 6.46 6.25 37 75.6 45
Bougainvillia 1.1 1.86 3.4 3.90 5.10 6.10 6.45 35 76.8 44
spectabilis
Ipomea palmate 0.73 1.63 2.43 3.46 5.23 5.7 5.60 44 64.3 53
Quadqualis indica 0.43 0.93 2.03 2.50 3.53 4.63 6.20 38 68.5 50
Callistemone 0.93 1.76 2.60 4.56 5.63 6.76 8.25 17 95.4 31
lanceolata
Control 1.1 1.5 3.2 4.75 6.4 9.5 10.0 139
LSD (0.05%) 5.6 12.3 5.2

compounds in higher plants has long been
recognized as an important factor in disease
resistance (Mahadevan, 1982). There are evidences
from earlier works that several plant species possess
antifungal and antibacterial properties (Bandara et
al., 1989; Srivastava and Lal, 1997; Maji et al.,
2005; Harlapur et al., 2007; Nduagu et al., 2008;
and Ashwani et al., 2011).
MATERIAL AND METHODS
Fresh green leaves of ten locally available plants
viz., Bauhinia variegate, Bougainvillia spectabilis,
Callistemone lanceolata, Clerodendron
splendensens, Hemelia patens, Ipomea palmata,
Nerium indicum, Polyalthia longifolia, Quadqualis
indica, Thevetia peruviana were collected from
different places of Aligarh. Pure cultures of R.
solani were isolated from the roots of
Pseuderanthemum atropurpureum. Potato Dextrose
Agar (PDA) was used as medium for sub-culturing
the R. solani.
Extraction and evaluation of the plant
materials
Fresh leaves of ten healthy plants were
collected and washed thoroughly with tap water
and air dried. One hundred grams of plant leaves
were ground using pestle and mortar by adding
equal amount (100 ml) of sterilized distilled water
(1:1, w/v). The pulverized mass was squeezed
through the cheese cloth and the extracts were
centrifuged at 10000 rpm for 5 – 10 minutes. The
supernatant was filtered using Whatman’s filter
paper no. 1 under aseptic conditions. It was stored
at 40C in air tight bottles for future use.
The effect of phytoextracts of different plant
species were tested in vitro by food poison
technique to know their inhibitory effect on the
growth of R. solani. The 5 ml of standard extract
was mixed with 50 ml of PDA and then autoclaved.
The autoclaved media along with extract was
transferred into sterilised petriplates aseptically. The
PDA plates containing the plant extracts were
inoculated aseptically with the pathogen by
transferring five mm diameter agar disc from the
fresh cultures. Three replications were maintained
for each treatment. The basal medium (PDA)
without any phytoextract served as the control. All
the inoculated Petri dishes were incubated at
25±1°C. The radial growth of the test fungus was
measured in all the treatments after three days and
compared with the control. The per cent inhibition
GANAIE and KHAN, Biotoxicity of Some Indigenously Available Plant Leaf Extracts Against Root-Rot Fungus
of fungal growth was estimated by using following
formula (Vincent, 1927):
I = C - T × 100
C Nerium indicum, Clariodendron splendensens and
Where, I = per cent inhibition
C = Colony diameter in control
T = Colony diameter in treatment
The data was recorded in triplicates and
subjected to statistical analysis and conclusions
were drawn on the basis of analysis of variance.
The calculated value of F was compared with the
tabulated values at 5% level of significance for an
appropriate degree of freedom.
RESULTS AND DISCUSSION
In the present study, the biotoxicity of the ten
locally available plant leaf extracts was evaluated
against root-rot fungus, Rhizoctonia solani. The
study involves the inhibitory effect of tested leaf
extracts on colony formation and sclerotia
production of R. solani. The data presented in Table-
1 reveals that minimum mycelial growth of R.
solani was observed in Polyalthia longifolia after
24 hours followed by Nerium indicum, Quadqualis
indica , Ipomea palmata , whereas, the maximum
growth of fungal colony formation after 24 hrs
was observed in Bouganvillia spectabilis and
Thevetia peruviana followed by Hemelia patens,
and Clariodendron splendensens. Time interval
played an important role in the development of
colony formation of R. solani. It was observed
that in general, growth of colony formation
increased with the increase in the time duration.
The maximum inhibitory effect of 44 per cent on
mycelial growth of R. solani after 164 hours of
inoculation was observed in the Petri plates
inoculated with leaf extracts of Ipomea palmate
followed by per cent reduction of 43, 38, 37, 35,
24, 17 and 13 due to inoculation of leaf extracts of
Bauhinia variegate, Quadqualis indica, Hemelia
patens, Bougainvillia spectabilis, Polyalthia
longifolia, Callistemone lanceolata and Nerium
indicum respectively.
It was also observed that a significant
reduction occurred in the sclerotia formation of
the test fungi R. solani inoculated with different
leaf extracts. The maximum number of sclerotia
occurred in the Petri plates which served as control
i.e without any inoculation. The minimum number
of 62.45 sclerotia production was observed in the
625
Petri plates inoculated with Bauhinia variegate
followed by Ipomea palmata, Quadqualis indica,
Hemelia patens, Bouganvillia spectabilis,
Polyalthia longifolia, Callistemone lanceolata,
Thevetia peruviana. In the corresponding
treatments the per cent reduction in the sclerotia
formation with respect to control was observed as
55, 53, 50, 45, 44, 35, 31, 28, 9 and 6 respectively.
The results of the present investigation are in
agreement with the earlier findings some
researchers who also observed the inhibitory effect
of different plant leaf extracts on the different fungi
( Bhat and Shukla 2001., Okigbo and Ogbonnaya
2006) . Different growth behavior of colony
formation and sclerotia production in different leaf
extracts may be due to sensitivity of the fungus
towards substances present in different leaf
extracts. The antifungal effect of the extracts
against mycelia growth varied with plant species
used. It was observed that in some cases the fungal
pathogen showed fast growth in the beginning but
the growth gradually declined at the end. This might
be due to the availability of favourable substances
in the beginning and their depletion with time and/
or auto toxicity developed by fungal spores.
It can be concluded from the present study
that out of the ten tested plant leaf extracts, eight
showed inhibitory effect against the root-rot
fungus, R. solani. Thus, these extracts can be
potentially useful for the control of root-rot caused
by test fungi R. solani and their inhibitory effect
will be evaluated under field conditions during the
future course of research.
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Received on 13-05-2016 Accepted on 18-05-2016