Identification and Evaluation of Antifungal Compounds from Botanicals

for the Control of Sugarcane Red Rot Pathogen, Colletotrichum falcatum

Keywords:
Sugarcane Red Rot, Colletotrichum falcatum, Psoralea corylifolia, Antifungal
Activity.
ABSTRACT:

Red rot is a devastating disease in sugarcane caused by fungus,
Colletotrichum falcatum. In this study, eighteen different botanicals were screened for
identifying effective antifungal compound against C. falcatum. Among the plants
screened, 15 per cent aqueous leaf extract of Psoralea corylifolia alone inhibited 100
per cent growth of both mycelium as well as spore germination under in vitro
conditions. The extract did not exhibit any inhibitory effect to the beneficial microbes
viz., Pseudomonas fluorescens, Bacillus megaterium and Gluconacetobacter
diazotrophicus which are normally used in sugarcane. The effective plant extracts
exhibiting 100 per cent antifungal activity was subjected to TLC, HPLC and GC-MS
analysis to identify the bioactive antifungal compound. It revealed the presence of 7H-
furo [3,2-G] (1) benzopyran-7-one as main bioactive compound which is thought to be
the intermediate of antifungal compound, 8 methoxypsoralen formed during
biosynthesis.
164-172 | JRA | 2013 | Vol 2 | No 1
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and
reproduction in all medium, provided the original work is properly cited.
www.jagri.info
Journal of Research in
Agriculture
An International Scientific
Research Journal
Authors:
Rajkumar D* and
Murugesan R.




Institution:
Department of Agricultural
Microbiology, Tamil Nadu
Agricultural University,
Coimbatore, 641 003,
Tamilnadu, India.





Corresponding author:
Rajkumar D.
















Email:



Web Address:
http://www.jagri.info/
documents/AG0044.pdf.


Dates:
Received: 11 May 2013 Accepted: 28 May 2013 Published: 13 June 2013
Article Citation:
Rajkumar D and Murugesan R.
Identification and Evaluation of Antifungal Compounds from Botanicals for the Control
of Sugarcane Red Rot Pathogen, Colletotrichum falcatum.
Journal of Research in Agriculture (2013) 2(1): 164-172
Original Research
Journal of Research in Agriculture
J
o
u
r
n
a
l

o
f

R
e
s
e
a
r
c
h

i
n

A
g
r
i
c
u
l
t
u
r
e

An International Scientific Research Journal


INTRODUCTION
Sugarcane is one of the most important cash
crops grown in India which contributes to the tune of
7.5 per cent of agricultural production of the country
(Viswanathan, 2010). But, sugarcane red rot disease
caused by fungus Colletotrichum falcatum is considered
as very serious disease and it was responsible for the
elimination of many elite sugarcane varieties such as Co
312, Co 658, Co 997, Co 1148, CoC 671 etc. Upto 100
per cent yield loss has been reported in severe
epiphytotics (Siddique et al., 1983). The use of chemical
fungicide bavistin is the common practice followed by
the sugarcane growers for the control of sugarcane red
rot. Practicing sett treatment with systemic fungicides
alone is not sufficient to protect such a long duration
crop. The use of synthetic fungicide leads to several
problems such as residue in food and feed, pathogen
resistance, toxicity to non target organism and
environmental pollution (Angelo et al. 2012). In addition
to these, elimination of soil born inoculum through
chemicals is difficult and costly. Development of
resistant varieties through breeding methods is a long
term endeavour. Therefore, with the increasing public
awareness of environmental safety and persistent
demand for ecofriendly products, we are forced to
produce quality products both for export and domestic
consumption. Hence, an alternative approach is the use
of botanicals in management of this disease which are
eco friendly in addressing the problem.

MATERIALS AND METHODS
Plants screened for antifungal study
Eighteen plants belonging to 14 families were
taken for screening antifungal activity against
Colletotrichum falcatum. All the plants were collected
from Medicinal Plant Section maintained at Botanical
Garden and agricultural fields of Tamil Nadu
Agricultural University, Coimbatore (Table 1). All the
experiments were done with three replications and the
data given are the mean of three replications. Since this
study is a preliminary qualitative screening study under
in vitro conditions quantitative data were not given and
so statistics was also not applied for the data.
Microbial cultures taken for study
Sugarcane red rot fungi, Colletotrichum falcatum
was obtained from Department of Plant Pathology,
Sugarcane Breeding Institute, ICAR, Coimbatore.
Thr ee ben ef i ci a l mi cr oor gani s ms vi z . ,
Pseudomonas fluorescens, Bacillus megaterium and
Gluconacetobacter diazotrophicus commonly applied for
sugarcane crop were obtained from Department of
Agricultural Microbiology, Tamil Nadu Agricultural
University, Coimbatore.
Preparation of plant extracts
Fresh plant parts of all botanicals were
thoroughly washed with running tap water followed by
rinsing twice with distilled water. The plant materials
were shade dried for 1 h. Weighed plant materials were
homogenized in mixer grinder with distilled water
(1:1 w/v). The slurry of plant extracts obtained was passed
through glass wool and the filtrate was then centrifuged at 5000
rpm for 10 min. Supernatant of botanicals thus obtained
were the standard aqueous plant extract solution (100
per cent). For methanol extract, weighed plant materials
were homogenized in mixer grinder with methanol
(1:3 w/v). The slurry samples collected in a beaker were
covered with aluminium foil and incubated in
refrigerator at 4C for 24 h. After incubation period,
slurry was filtered using glass wool and subsequently
with Whatman No. 1 filter paper. The clear filtrate was
allowed to evaporate in petriplates and the residues were
dissolved in distilled water to obtain standard methanol
plant extract solution of all botanicals (Biri Singh, 2004).
In vitro testing of plant extracts against test pathogens
The antifungal effects of all the plant extracts
were screened by poison food technique (Horsefall,
1956). An appropriate amount of plant extracts were
added to sterilized potato dextrose agar (PDA) medium
165 Journal of Research in Agriculture (2013) 2(1): 164-172
Rajkumar and Murugesan , 2013
to obtain the desired concentration of 25 per cent. Then
the medium containing plant extracts were kept in water
bath at 45C for 60 min to eliminate bacterial contamination
(Martinez et al., 1990) and plated after cooling it to luke
warm temperature. C. falcatum mycelial disc of 9 mm
diameter was taken from 3-5 days old culture plate and
placed to the centre of each plates and incubated at
28C. After 5 days of incubation the diameter of
mycelial colony was measured and the per cent
inhibition for all botanicals was calculated by using the
formula (Vincent, 1927).
Where, C and T were mycelial growth (cm) of test
fungi in control and treatments, respectively. All treatments
were done in triplicates and average values were taken for
interpretation.
Minimum inhibitory concentration (MIC)
Minimum inhibitory concentration is the lowest
concentration at which 100 per cent inhibition of
mycelial growth is obtained. Among the different
botanicals screened, P. corylifolia aqueous leaf extract,
which exhibited effective inhibition over C. falcatum,
was tested at different concentrations (5, 10, 15 and
20 per cent) to find out MIC by poison food technique.
Evaluation of the effect of Psoralea corylifolia extract
on the spore germination of Colletotrichum falcatum
P. corylifolia extract of different concentrations
(5, 10, 15 and 20 per cent) were prepared in tubes for
checking its effect on spore germination of C. falcatum.
A loopful of conidia from sporulated culture in oatmeal
enriched PDA medium were transferred into extract
containing tubes. From each extract concentration, 100
l was taken into the cavity slides along with the control
(without extract). These slides were kept in a moist
chamber for the period of 24 h for germination of spores
and the effect of P. corylifolia extract on spore
germination was determined by counting germinated
spores under microscope and it was compared with the
control (Jayakumar, 2007).
Evaluation of the effect of P. corylifolia extract on
growth pattern of beneficial organisms
In vitro testing of the P. corylifolia plant extract
was carried out on beneficial microflora to assess its
influence on their growth. Twenty four hour old cultures
of Pseudomonas fluorescens, and Bacillus megaterium
were streaked on the nutrient agar medium
cont ai ni ng speci fi c pl ant ext r act a n d
Journal of Research in Agriculture (2013) 2(1): 164-172 166
Rajkumar and Murugesan , 2013
Table 1. List of medicinal plants screened for its antifungal activity
S.No Family Botanical name Parts used
1. Amaranthaceae Achyranthes aspera Leaf
2. Malvaceae Abelmoschus esculentus Leaf
3. Papilionaceae Abrus precatorius Leaf
4. Annonaceae Annona squamosa Leaf
5. Nyctaginaceae Bougainvillea spectabilis Leaf
6. Asclepiadaceae Calotropis gigantea Leaf
7. Verbenaceae Clerodendrum inerme Leaf
8. Caesalpiniaceae Caesalpinia bonduc Bark
9. Sapindaceae Cardiospermum halicacabum Leaf
10. Asteraceae Eclipta alba Leaf
11. Verbenaceae Lantana camara Leaf
12. Moringaceae Moringa oleifera Leaf
13. Meliaceae Melia azadirachta Leaf
14. Fabaceae Prosopis juliflora Leaf, Stem
15. Papilionaceae Psoralea corylifolia Leaf, Stem
16. Apiaceae Trachyspermum ammi Leaf, Stem
17. Verbenaceae Vitex negundo Leaf
18. Asteraceae Wedelia urticaefolia Leaf
Inhibition (%) =
(C T)
X 100
C


Gluconacetobacter diazotrophicus was streaked on the
potato dextrose agar medium containing plant extract and
suitable control (without plant extracts) was also maintained.
The plates inoculated with Pseudomonas fluorescens, and
Bacillus megaterium were incubated for two days whereas
Gluconacetobacter diazotrophicus plates were incubated
for five days. The growth of the organism was observed
visually, compared with that of the control and given the
remark as for no growth, + for very less growth, ++ for
moderate growth and +++ for growth equivalent to control.
Control indicates petri plates containing PDA medium
without plant extracts. It was taken to check whether there is
any difference in the growth pattern of beneficial microbes
inoculated in PDA medium with plant extracts and without
plant extracts.
Identification of the fungitoxic principles through
TLC, HPLC and GC-MS analysis
Thin layer chromatographic study was carried
out to identify the nature of active principle in the
effective plant extracts. The solvent systems used were
ethyl acetate: methanol: ammonia solution (17:5:1),
chloroform: acetic acid (9:1) and ethyl acetate: formic
acid: acetic acid: water (100:11:11:27) for alkaloids,
phenols and flavonoids, respectively. Dragendroffs
r ea g en t , Fol i n - Ci oc a l t e u r ea g en t a n d
Diphenylboryloxyethylamine (1 per cent) with 5 per cent
polyethylene glycol 4000 were used as spray reagents.
The dried TLC plate was exposed to UV light (254 nm)
and observed for fluorescent spots. The compounds
separated in TLC were checked for its antifungal activity
against C. falcatum by poison food technique. The active
compounds which exerted antifungal activity were
subjected to HPLC analysis.
HPLC analysis
HPLC shimadzu model L 6200 (Varian) diode
array detector in combination with Beckman ultrasphere
supelco ODS column (250 x 4.6 mm) was used for
analysis. The analysis was performed at a flow rate of
0.35ml min
-1
using Methanol: Water (40:60) as mobile
phase. The UV detector was set at 260nm wavelength for
psoralen (Murali et al. 2002). Psoralen standard solutions
(0.5, 1.0, 1.5 and 2.0 percent) were prepared and 20 l of
the solution was injected to record the retention time.
The individual peaks separated in HPLC was collected
through fraction collector and evaluated for its antifungal
activity by poison food technique. The peak which
exerted antifungal activity was again subjected to
GC-MS analysis.
GC-MS analysis
GC-MS JEOL GC mate Hitachi model L 6200
(Varian) photo array detector in combination with
Beckman Ultrasphere supelco ODS column (250 x 4.6
mm) was used for analysis. The analysis was performed
at a flow rate of 0.35ml min
-1
. The UV detector was set
at 260nm wavelength for Psoralen. The peaks obtained in
chromatogram were analysed for its mass spectrum to
identify the compounds by comparing its molecular
weight with the compounds present in MS library.

RESULTS AND DISCUSSION
Antifungal activity of different botanicals
Among 18 plants tested for the antifungal
activity against C. falcatum, 15 per cent aqueous leaf
extract of P. corylifolia inhibited 100 per cent growth of
both mycelium as well as spore germination under
laboratory conditions (Plate 1and 2). None of the plant
extracts other than P. corylifolia were able to exhibit
comparatively appreciable inhibition over C. falcatum.
The presence of antifungal compounds in medicinal
plants has long been recognized as an important factor to
disease resistance (Mahadevan, 1982; Kurucheve et al., 1997).
Such compounds, being biodegradable and selective in
their toxicity are considered valuable for controlling
different plant diseases (Singh and Dwivedi, 1987).
Rajendra Prasad et al., (2004) reported the antifungal
activity of extracts obtained from P. corylifolia seeds
against dermatophytic fungi such as Trichophyton
rubrum, Trichophyton mentagrophytes, Epidermophyton
167 Journal of Research in Agriculture (2013) 2(1): 164-172
Rajkumar and Murugesan , 2013
floccosum and Microsporum gypseum. Mazzuca et al.
(2003) also observed antifungal activity of its seed
extracts against some fungi causing infection in humans.
Sindhan et al., (1999) reported that 10 per cent leaf
extracts (aqueous) of Allium cepa, Zingiber officinale,
Azadirachta indica, Allium sativum, Mentha arvensis,
Eucalyptus globulens, Ocimum sanctum and Datura alba
were effective in inhibiting the mycelial growth of R.
solani to the extent of 40 60 per cent. The inhibitory
effect of the extract of Prosopis juliflora leaves was
studied against Gloeosporium piperatum (Gomathi and
Kannabiran, 2000) and against R. solani (Girija Shankar
and Thayumanavan, 2000). It is already known from the studies of
Khursheed et al. (1986) and Aqeel Ahmad et al., (1997) that
juliflorine, an alkaloid obtained from Prosopis juliflora
leaves, is the actual compound which is responsible for
antifungal activity. Available reports on P. corylifolia
suggest that the antifungal activity has been tested only
in seeds, not with other plant parts. But the present study
indicated that antifungal principles are also present in
leaves of P. corylifolia which exhibited more antifungal
activity. However, the results of the present study
Journal of Research in Agriculture (2013) 2(1): 164-172 168
Rajkumar and Murugesan , 2013
Plate 1. Effect of Psoralea corylifolia leaf extract on mycelia growth of C. falcatum
Control
15 % leaf extract
Control
Plate 2. Effect of Psoralea corylifolia leaf extract on spore germination of C. falcatum
15 % leaf extract


pertaining to the antifungal activity of P. juliflora are
fully in corroboration with the earlier reports as
discussed above.
Compatibility studies of effective concentration
(15%) of P. corylifolia leaf extract against beneficial
bacteria
In vitro testing of the effective plant extract was
carried out on beneficial microflora to assess whether
plant extract is inhibitory or non inhibitory to their
growth. The effective concentration (15 per cent) of leaf
extract of Psoralea corylifolia was taken for testing the
compatibility assessment with beneficial bacteria
Pseudomonas fluorescens, Bacillus megaterium, and
Gluconacetobacter diazotrophicus. The extract showed
no inhibitory effect and found to be compatible with all
beneficial microflora tested (Table 2). The results in the
present study were not in agreement with that of
Parimala and Sabitha Doraisamy (2000) who examined
the compatibility of six plant extracts viz., Azadirachta
indica, Abrus precatorius, Vitex negundo, Nerium
oleander and Catharanthus roseus for Trichoderma
viride strains (MNT 5 and MG 6), Pseudomonas
fluorescens and Bacillus subtilis, but found that these
plant extracts were non-compatible to the tested
antagonists. This may be due to the reason that the tested
plant extracts may be possessing different type of
compounds which are of inhibitory nature to the
beneficial antagonists. Literature on compatibility testing of
plant extracts with beneficial microflora viz., Bacillus
megaterium, Pseudomonas fluorescens and
Gluconacetobacter diazotrophicus had not been reported
earlier.
Identification of antifungal compound from
Psoralea corylifolia extract
Different chromatographic studies such as TLC,
HPLC and GC-MS were carried out to identify the
specific compounds in P. corylifolia responsible for
antifungal activity against C. falcatum. In TLC study,
three different solvent systems were used for identifying
alkaloids, phenols and flavonoids. Among which the
flavonoid compounds which were well separated in the
ethyl acetate: formic acid: acetic acid: water
(100:11:11:27) solvent system alone exhibited antifungal
activity against C. falcatum which was confirmed by
poison food technique. The flavonoid compounds
separated in TLC were subjected to HPLC analyses, in
which it revealed three major peaks upon separation
(Figure 1). Among these, the compound showing second
peak exerted antifungal activity against C. falcatum which
was again confirmed by poison food technique. The
effective peak was collected through fractional collector
in HPLC and it was subjected to GC-MS analysis. GC-
MS analysis revealed five major peaks upon separation
(Figure 2 ) and these peaks were identified as, 2-[1-
cyclohexylethyl)carbamoyl] methyl ester propanoic acid (RT
2.551 min.), N-methyl-2-methoxy-1-propanamine (RT 3.360
min.), 2-propoxy ethanol (RT 3.451 min.), 1,2-
dimethylbenzene (RT 4.570 min.) and 7H-furo[3,2-G](1)
benzopyran-7-one (RT 25.062 min.). There is no report about
the fungicidal effect of 7H-furo[3,2-G] (1) benzopyran-7
-one. However, there are reports about the antifungal
nature of 9-methoxy-7H-furo [3,2-G](1) benzopyran-
7-one / 8-methoxypsoralen known to present in
P. corylifolia. The antifungal activity of
169 Journal of Research in Agriculture (2013) 2(1): 164-172
Rajkumar and Murugesan, 2013
Table 2. Compatability of effective leaf extract with beneficial organisms
S.No Name of Organism Control 15% extract
1 Bacillus megaterium (phosphorus solubilizer) +++ +++
2 Pseudomonas fluorescens (biocontrol agent) +++ +++
3 Glucanoacetobacter diazotrophicus (nitrogen fixer) +++ +++
+ - low compatible, ++ - Moderately compatible, +++ - Highly compatible
8-methoxypsoralen was studied already against
Fusarium oxysporum (Oliva et al., 2003) and against
dermatophytic fungi (Kundsen, 1980). Hence, it can be
presumed that resulting fungitoxicity of P. corylifolia
may be due to 8-methoxypsoralen, a furanocoumarin
which might be formed via 7H-furo [3,2-G](1)
benzopyran-7-one during biosynthesis. Antifungal effect
of 8-methoxypsoralen is reported due to its
photosensitizing potency (Young and Barth, 1982) and its
binding with nuclear DNA (Bredberg et al., 1977).
Rajkumar and Murugesan , 2013
Journal of Research in Agriculture (2013) 2(1): 164-172 170
Figure 2. Mass spectrum of the compound
Minutes
0 2 4 6 8 10 12 14 16 18 20 22 24
m
A
U
0
500
1000
1500
2000
2500
3000
3500
m
A
U
0
500
1000
1500
2000
2500
3000
3500
0
.
1
9
2


4
8
7


1
9
5
0
.
5
3
3


4
3
9


1
9
4
1
.
1
4
1


1
4
5
1


1
9
5
1
.
3
3
3


2
5
5


2
0
6
1
.
4
5
1


2
9
0


1
9
5
3
.
0
2
9


1
7
9
3
6
2
8


2
0
1
5
.
1
3
1


3
6
5
4
0
1
4
5


2
4
9
6
.
7
5
2


9
6
1
6
5
7
3
8


2
5
9
1
2
.
1
2
8


3
6
2
7
9
5
8
5


2
3
9
1
6
.
0
9
6


1
2
5
7
3
0


1
9
1
1
7
.
7
3
9


6
2
8
6
7


1
9
4
1
8
.
7
7
3


2
1
6
9


1
9
1
1
9
.
4
0
3


3
5
4
5


1
9
1
1
9
.
5
9
5


1
9
3
4
9


2
0
3
2
4
.
1
3
9


2
4
2
8


1
9
4
2
4
.
3
0
9


1
6
4
7


1
9
3
2
4
.
7
7
9


3
8
8


1
9
5
1: 262 nm, 8 nm
Psoralen extract-r2
run2
Retention Time
Area
Lambda Max
Figure 1. HPLC of the compound
CONCLUSION
Various chemical fungicides have been
recommended for the control of different diseases.
However, their use cause several adverse effects and
have posed various problems like residues in feed and
food, pathogen resistance, toxicity to non-target
organisms, environmental pollution in different
agricultural ecosystems and hence it has become
necessary to use eco-friendly formulations which can fit
into integrated disease management. This study screened
different botanicals for the control of C. falcatum,
sugarcane red rot pathogen. It was found that the 15 %
leaf extract of P. corylifolia possess strong fungitoxic
properties under in vitro conditions worth exploiting for
the biomanagement of the sugarcane red rot disease.
Further studies are needed to test the fungitoxic effect
under field conditions, their thermostability, stability to
storage and also their phytotoxicity towards the host plant.
This study also confirmed the compatibility of fungitoxic
principles of plants with beneficial microbes. Therefore,
the present study augurs well for the establishment of the
fact that they should possess both the properties ie.,
toxicity to pathogenic fungi and safety to beneficial
microbes for their application in sustainable agriculture.

REFERENCES
Angelo Stefani Jr, Joanna DArc Felcio, Mara M. de
Andra. 2012. Comparative Assessment of the Effect of
Synthetic and Natural Fungicides on Soil Respiration,
Sensors. 12 (3): 3243-3252.

Ahmad A., Ahmad VU, Khalid SM, Ansari FA, Khan
KA. 1997. Study on the antifungal efficacy of Juliflorine
and a benzene-insoluble alkaloidal fraction of Prosopis
juliflora, Philipp. J. Sci., 126(2), 175 - 182.

Biri Singh. 2004. Screening of antimicrobials from
medicinal plants and assessing their influence on certain
phytopathogenic fungi and beneficial microbes. M.Sc.,
Thesis, submitted to TNAU.Coimbatore.

Bredberg A, Lambert B, Swanbeck G, Thyresson -
Hok M. 1977. The binding of 8-methoxypsoralen to
nuclear DNA of UV irradiated human fibroblasts in
vitro. Acta Derm. Venerol., 57(5): 389-3891.

Girija Shankar, Thayumanavan B. 2000. Antifungal
compounds from Prosopis juliflora, Pongamia pinnata
and Lawsonia inermis. M.Sc (Biotechnology)., Thesis,
submitted to TNAU, Coimbatore.

Gomathi V, Kannabiran B. 2000. Inhibitory effects of leaf
extracts of some plants on the anthracnose fungi infecting
Capsicum annum. Indian Phytopath., 53(3):305-308.

Horsefall JG. 1956. Principles of Fungicidal Action.'
Chronica Botanica Co., Waltham, Mass.

Jayakumar T. 2007. Studies on the influence of
screened botanicals on the control of red rot disease
(Colletotrichum falcatum) of sugarcane and their
compatibility with beneficial microbes. M.Tech
(Microbial Technology). Department of Agricultural
Microbiology. Thesis, submitted to TNAU. Coimbatore.

Khursheed AK, Arshad HF, Viqaruddin A, Sabiha Q,
Sheikh AR, Tahir SH. 1986. In vitro studies of
antidermatophytic activity of juliflorine and its screening
as carcinogen in Salmonella / microsome test system.
Arzneimittelforschung., 36(1): 17-19.

Kundsen EA. 1980. In vitro studies on the antifungal
effect of PUVA. Acta. Derm. Venereol., 60(5): 452-456.

Kurucheve V, Gerard Ezhilan J, Jayaraj J. 1997.
Screening of higher plants for fungitoxicity against
Rhizoctonia solani in vitro. Indian Phyopathol., 50 (2):
235-241.

Mahadevan A. 1982. Biochemical aspects of plant
disease resistance. Part I. Performed inhibitory
substances- Prohibitins. Today and tomorrows, Printers
and Publishers, New Delhi.

Rajkumar and Murugesan , 2013
171 Journal of Research in Agriculture (2013) 2(1): 164-172
Martinez MC, Sanchez-Montero JM, Sinisterra JV,
Ballesteros A. 1990. New insolubilized derivatives of
ribonuclease and endonuclease for elimination of nucleic
acids in single cell protein concentrates. Biotechnol.
Appl. Biochem., 12(6): 643-52.

Mazzuca M., Kraus W, Balzaretti V. 2003. Evaluation
of the biological activities of crude extracts from
patagonian prosopis seeds and some of their active
principles. J. Herb Pharmcother., 3 (2), 31-37.

Murali B, Amit A, Anand MS, Venkataraman BV.
2002. An HPLC method for simultaneous estimation of
psoralen, bakuchicin and bakuchiol in Psoralea
corylifolia. J. Natural Remedies, 2 (1): 76-80.

Oliva A, Meepagala KM, Wedge DE, Harries D, Hale
AL, Aliotta G, Duke SO. 2003. Natural fungicides from
Ruta graveolens L. leaves, including a new quinolone
alkaloid. J. Agric. Food Chem., 51(4):890-896.

Parimala K, Sabitha Doraisami. 2000. Effect of
botanicals against groundnut late leaf spot and rust
diseases. M.Sc. (Ag.) Thesis, Dept. of. Plant Pathology.
Tamil Nadu Agricultural University, Coimbatore.

Rajendra Prasad N, Anandi C, Balasubramanian S,
Pugalendi KV. 2004. Antidermatophytic activity of
extracts from Psoralea corylifolia (Fabaceae) correlated
with the presence of a flavonoid compound.
J. of Ethnopharmacol., 91(1): 2124.

Siddique, Mirza M, Khanzada MI, Maazulla Khan.
1983. Incidence and effects ofsugarcane red rot caused
by Colletotrichum falcatum in northern pakisthan.
Pakistan J. Agric. Res., 4(3): 208.

Sindhan GS, Hooda I, Parashar RD. 1999. Evaluation
of plant extracts for the control of powdery mildew of
pea. J. Mycol. Pl. Pathol., 29(2):257-258.

Singh PK, Dwivedi RS. 1987. Effect of oils on
Sclerotium rolfsii causing root rot of barley. Indian
Phytopathol., 40 (2): 531-533.

Vincent JM. 1927. Distortion of fungal hyphae in the
presence of certain inhibitors. Nature, 159 (4051): 850.

Viswanathan R. 2010. Plant Disease: Red rot of
sugarcane. Anmol Publishers, New Delhi, 305.

Young AR, Barth J. 1982. Comparative studies on the
photosensitizing potency of 5-methoxypsoralen and
8-methoxypsoralen as measured by cytolysis in
Paramecium caudatum and Tetrahymena pyriformis and
growth inhibition and survival in Candida albicans.
Photochem. Photobiol., 35(1):83-88.
Rajkumar and Murugesan , 2013
Journal of Research in Agriculture (2013) 2(1): 164-172 172
Submit your articles online at www.jagri.info

Advantages
Easy online submission
Complete Peer review
Affordable Charges
Quick processing
Extensive indexing
You retain your copyright


submit@jagri.info

www.jagri.info/Submit.php.