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B - 683
Research Article Microbiology





nternational !o"rnal of #harma and Bio $ciences
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A%TM+R,BA- A+T.T/ ,0 $1+,%2AR/ M1TAB,-T1$ 0R,M
1%2,#3/T+ 04%5 $,-AT12 0R,M %1R4M ,-1A%21R -.

RAMESHA. A, SUNITHA.V. H, AND C. SRINIVAS
*


Department of Studies in Microbiology & Biotechnology, Jnanabharathi Campus, Bangalore University,
Bangalore !" "!, #arnata$a, %ndia&

ABSTRACT

Endophytic fungi found inside the plant tissue are endosymbionts, protecting their host from
pests, pathogens, etc. Twenty eight endophytic fungi were isolated from different parts of
Nerium oleander L., out of which, 54% of isolates were from flower, 3% from stem and !%
from leaf parts. Thirty si" percent of the isolates showed antimicrobial acti#ity against tested
pathogens. The potential isolates such as Fusarium semitectum $%of&3', Colletotrichum
gloeosporioides $%of&!', Alternaria alternata $%of&(' and )ycelia *terilia sp.+ $%os&,' were
sub-ected to the production and e"traction of secondary metabolites. .ll the four fungal
e"tracts inhibited Staphylococcus aureus and Bacillus cereus at /0 1g2mL $)34'. E"tracts
of C. gloeosporioides and )ycelia *terilia sp.+ showed acti#ity against Pseudomonas
aeruginosa at )34 of /0 1g2mL. The growth of Escherichia coli was suppressed by all the
tested e"tracts at )34 of /0 1g2mL e"cept F. semitectum. A. alternata 5 )ycelia *terilia
sp.+ e"tracts were acti#e against Salmonella typhimurium at /0 1g2mL. The growth of
Candida albicans was inhibited by )ycelia *terilia sp.+ at /0 1g2mL. The 6ones of inhibition
were statistically significant with respecti#e positi#e controls.

Keywords: Nerium oleander L.,Endophytic fungi, Antimicrobial activity, Secondary metabolites,
Minimum inhibitory concentration.











C. SRINIVAS
2epartment of $t"dies in Microbiology 6 Biotechnology7 !nanabharathi +amp"s7
Bangalore 4ni8ersity7 Bangalore 9 )6& &)67 :arnata;a7 ndia.
srini8asb"b<gmail.com

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B - 68=
INTRODUCTION

Endophytic fungi inhabit the host plant,
primarily within the aerial tissues, without
causing ad#erse effects. They are associated
symbiotically, such that endophytes draw
nutrients and protection from the host, but
contribute to effecti#e host defence against
pathogens, herbi#ores or abiotic stress
/0,/
.
.ppro"imately, a million species of endophytic
fungi are estimated to be present within the
global flora

. These endophytic fungi are rich

sources of secondary metabolites, which find a
wide range of applications in pharmacology as
well as in agriculture. They are absolutely
needless for the producers and do not in#ol#e
in their life cycle. The crystalline compound
mycophenolic acid isolated from Penicillium
glaucoma was considered as the first microbial
secondary metabolite, disco#ered in +(, by
7osio
++
. )ost important characteristic features
of secondary metabolites are their uni8ue
chemical structures, fre8uent occurrence and
biological acti#ity
4
. 9iological acti#ity will be
used generally for interactions between
chemicals and molecular targets of li#ing
organisms
++
. :ungal endophytes could also
produce metabolites similar to, or with more
acti#ity than that of their respecti#e hosts
/4
.
Therefore, it is belie#ed that search for no#el
compounds should be directed towards
endophytic fungi for medicinal purpose and to
sa#e the mass utili6ation of plants to produce
secondary metabolites. %aturally produced
bioacti#e metabolites from endophytic fungi
include al;aloids, ben6opyranones,
ben6o8uinones, fla#onoids, isocoumarin,
lignans, phenol and phenolic compounds,
phenylpropanoids, steroids, terpenoids,
tetralones, "anthones, and other compounds
/(
. Nerium oleander L. is an e#ergreen shrub
$or small tree' that grows to appro"imately , m.
The stic;y late" of the stem is highly poisonous
to humans, pets, li#estoc; and birds due to the
presence of cardiac glycosides, mainly
oleandrin. 3ngestion causes nausea, #omiting,
cardiac arrhythmias, hypotension $low blood
pressure' and death. 3ts sap has been used as
rat poison
+,+/
. The plant is also pro#en to ha#e
medicinal #alue li;e cardiotonic, emetic,
antibacterial, anti&inflammatory, anti&
nocicepti#e acti#ity and is used to treat scabies
(,+3
. <aluable compounds li;e cardiac steroid,
arabinogalactan and cardenolides are reported
to be present in this plant
+5,+,//
. Endophytic
fungal isolates of N. oleander possessing
antio"idant, "anthine o"idase inhibitory and
antimicrobial acti#ity was reported by =u&>ang
et al
30
. The current in#estigation aimed at
isolation of endophytic fungi from #arious parts
of N. oleander L. and to e"plore their
antimicrobial potential against human
pathogens.

MATERIALS AND METHOD

(I) Sources of endophytic fungi
?ifferent plant parts of N. oleander L. were
collected $e"cised using a sterile ;nife' from
?han#antri #ana located in )ariyappanapalya,
?epartment of forestry, 7o#ernment of
@arnata;a, 9angalore, Alant samples were
brought to the laboratory in sterile polythene
bags. Berbarium of plant sample $with the
accession :CLBT 4oll. %o. !40,3' was
deposited in 3nstitute of .yur#eda and
3ntegrati#e )edicine $:CLBT', %o. !42/,
Dara;abande ;a#al, .ttur $Aost' <ia >elahan;a,
9angalore& 5,00,4.

(II) Isolation of endophytic fungi
?ifferent parts of fresh healthy Nerium oleander
L. plant were cut into small pieces $5 mm E /
mm' using sterile blade, washed with sterile
distilled water. *urface sterili6ation was done by
immersing in 4% sodium hypochlorite solution
for 0 *ec followed by !0% ethanol treatment
for 5 *ec, thoroughly washed with sterile
distilled water
/5
and blot dried between sterile
paper towels. The surface sterili6ed samples
were placed on potato de"trose agar $A?.'
plates amended with 50 mg2L tetracycline to
Int J Pharm Bio Sci 2013 Jan; 4(1): (B) 683 - 693


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B - 68)
suppress the bacterial growth and incubated at
/(
F
4 to 30
F
4 for / to 3 days. The hyphal tip of
endophytic fungi growing out from the plant
tissue was transferred to fresh A?. plates
amended with 50 mg2L tetracycline. .fter
incubation at 30
F
4 for ! to +4 days, purity of the
culture was determined by colony morphology
/,
. 4oloni6ation rate, e"pressed in percentage
was calculated as the number of fungal isolates
from each part of the plant di#ided by total
number of endophytic fungi isolated from the
plant.

(III) Identification of endophytic fungi
The endophytic fungi were identified based on
the cultural characteristics, morphology of the
fruiting bodies and spores, using standard
manuals
3,,,/!
.

(IV) Screening for antimicrobial activity
Endophytic fungi isolated from the medicinal
plant N. oleander L. were screened for
antimicrobial acti#ity against the human
pathogenic bacteria $Staphylococcus aureus
%43) %o. /0!, Bacillus cereus %43) %o.
/+0,, Pseudomonas aeruginosa %43) %o.
//00, Escherichia coli %43) %o. //5,,
Salmonella typhimurium %43) %o. /50+' and
yeast $Candida albicans %43) %o. 34!+'
procured from %43), %4L, Aune. .gar plug
methodG 4ylindrical pieces were cut out from
well grown culture of the endophytic fungal
strains on A?.. The bloc;s were placed on the
Aetri dishes deep inoculated with a fi"ed
amount of test micro&organisms grown in
%utrient agar $%.' medium for bacteria and
*abouraud de"trose agar medium $*?.' for
yeast $+0
,
cells2mL'. The plates were ;ept for
+/ hours at /H(F4 for the antimicrobial
compound diffusion and thereafter they were
incubated for the growth of test&micro&
organisms at /5F4 for >east and 3!F4 for
bacteria. The antimicrobial acti#ity was
measured in the form of diameter 6one
inhibition after /4 hrs for bacteria
$Staphylococcus aureus, Bacillus cereus,
Pseudomonas aeruginosa, Escherichia coli,
Salmonella typhimurium' and 4( hrs for yeast
$Candida albicans'
5
$:ig 3'.

(V) Production and extraction of secondary
metabolites
Endophytic fungi e"hibiting potential
antimicrobial acti#ity were sub-ected to the
production of secondary metabolites. The fresh
mycelia grown on A?. plates at /(F4 for 3H,
days was inoculated into 500 mL flas;s
containing /00 mL of potato de"trose broth
medium $?e"trose, /0 gI e"tract of /00 g
potatoI +000 mLI autocla#ed at +/+F4 for /0
min', followed by incubation at /(F4 with +40
re#2min for +5 days. The culture broth of
endophytes was filtered using =hatmann %o. +
filter paper to remo#e mycelium. The secondary
metabolites from %of&3, %of&! and %of&( were
e"tracted from the culture filtrates using e8ual
#olume of ethylacetate. The red pigment
produced by %os&, isolate was e"tracted with
ethylacetate after acidification with acetic acid at
pB&3. The e"tracts were e#aporated under
#acuum at 50F4 and dried.

(VI) Antimicrobial activity of ethylacetate
extracts of the endophytic fungi by agar well
diffusion method
. fi"ed amount of test micro&organisms
$+0
,
cells2mL' were inoculated in Aetri dishes
containing %. for bacteria and *?. for yeast.
The crude ethylacetate e"tracts dissol#ed in
?)*J at different concentrations
$+0Kg2mL,/0Kg2mL, 40 Kg2mL, ,0 Kg2mL, (0
Kg2mL, +00 Kg2mL' were added into the 5mm
diameter well made inoculated Aetri dishes. The
cultures were ;ept for /4 hours at /H(L4 for the
antimicrobial metabolite diffusion and thereafter
they were incubated at the appropriate
temperature for the growth of test micro&
organisms. The 6one of inhibition was
measured in mm
/
$:ig 4'. $<3' *tatistical
.nalysisG The statistical analysis was analy6ed
by Two way .%%J<. with *A** ++.5, mean
#alues of the triplicates was compared
according to ?uncan )ultiple Cange Test
$?)CT' at p M 0.05.

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B - 686
RESULTS AND DISCUSSION

(1) Isolation and Identification of endophytic
fungi
Twenty eight different strains of endophytic
fungi were isolated from 300 tissue segments
$+00 segments from each stem, flower and leaf'
of N. oleander L. The coloni6ation and isolation
rates of endophytic fungi from flower $54%'
were higher than that of stem $3%' and least in
leaf $!%' parts, whereas =u&>ang et al.,
30

described more endophytic fungi in lea#es than
in stems $:ig+'. 3n the present study, Curvularia
sp., Fusarium spp. was found both in stems
and flower parts. Drechslera sp., Cladosporium
spp., hielavia terricola, three )ycelia sterilia
$non&sporulating strains' and two unidentified sp
found in stem segments. To the best of our
;nowledge, before this there are no reports of
hielavia terricola as an endophyte. =u&>ang et
al.,
30
reported Chaetomium sp., Cladosporium
sp., Colletotrichum sp., and orula sp. in the
stems of N. oleander L. 3n the present
in#estigation orula sp., Alternaria alternata,
Colletotrichum sp., Cylindrocephalum sp.,
Aspergillus sp., Penicillium sp., Cochliobolus
sp., and Chaetomium sp. were isolated from
flowers. Bipolaris sp. and )ycelia sterilia sp.5
were isolated from the lea#es of Nerium
oleander L. =u&>ang et al.,
30
reported that
)ycelia sterilia sp. $50% relati#e fre8uency'
were found predominantly in the lea#es along
with an Ascomycete sp. $TableG+'. Drechslera
sp., Cladosporium spp, Curvularia sp.,
Chaetomium sp., Colletotrichum sp., were
commonly found endophytes in medicinal plants
+0,+(
. Colletotrichum gloeosporioides was
reported from Plumeria acuti!olia belonging to
Apocynaceae
+,
.

() Screening for Antimicrobial activity
.mong /( endophytic fungi screened for
antimicrobial acti#ity, 3,% of isolates showed
acti#ity, ,4% of them did not possess any
acti#ity against any test pathogens. *i" isolates
showed potential inhibition against S. aureus
and se#en isolates inhibited the growth of B.
cereus. Jnly )ycelia *terilia sp.+ $%os&,'
suppressed the growth of C. albicans and two
strains were effecti#e against P. aeruginosa.
:i#e isolates e"hibited inhibition 6one against
gram&negati#e organism E. coli and S. typhi.
$Table.+'. .mong the tested organisms, four
endophytic fungi identified as Fusarium
semitectum $%of&3', Alternaria alternata $%of&!',
Colletotrichum gloeosporioides $%of&(' and
)ycelia sterilia sp.+ $%os&,' were found to be
more promising antimicrobial strains for
production and e"traction of secondary
metabolites. The antimicrobial property of
)ycelia sterilia sp.+ was due to production
e"tracellular red pigment. )orphological
identification of these four potential endophytic
fungi was further confirmed by .ghar;ar
research institute, Aune, 3ndia.

(!) Antimicrobial activity of crude extract by
agar well diffusion method"
The crude e"tract of the potential endophytic
fungal isolates, e"hibited a broad spectrum of
antimicrobial acti#ity against the pathogens,
when compared with that of standard positi#e
control tetracycline for bacteria and flucona6ole
for yeast. The 6one of inhibition ranged from
+.,! to /(.,! mm at concentrations of +0H+00
1g2mL. .ccording to Cios and Cecio,
/+
e"tracts
of natural origin showing antimicrobial acti#ity
abo#e +00 1g2mL concentration should be
a#oidedI hence we used the crude e"tract at
concentrations +0H+00 1g2mL. .ll the four
fungal e"tracts showed )34 at /0 1g2mL
against S. aureus and B. cereus, howe#er,
there was no inhibition at concentration below
/0 1g2mL. =hereas, =u&>ang et al.,
30
reported
)34 of endophytic fungal crude e"tract isolated
from N. oleander L. at range of +./5H+0 mg2mL
for S. aureus, 5H/5 mg2mL for B. cereus, 5H
+/.5 mg2mL for E.coli. This shows that
endophytes of N. oleander L., from different
ecological and geographical conditions e"hibit
8ualitati#ely and 8uantitati#ely #aried
antimicrobial acti#ity
+!
. 3n this study, A.
alternata and )ycelia sterilia sp.+ e"hibited )34
at /0 1g2mL against P. aeruginosa. A. alternata,
C. gloeosporioides and )ycelia sterilia sp.+
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B - 68(
inhibited E. coli at the same concentration,
whereas, only )ycelia *terilia sp.+ inhibited C.
albicans at )34 /0 1g2mL. C. gloeosporioides,
)ycelia sterilia sp.+ inhibited S. typhi at )34 /0
1g2mL. Bowe#er, none of the other crude
e"tracts tested showed inhibition e#en at +00
1g2mL. :ernandes
!
reported antimicrobial
acti#ity of e"tract of A. alternata isolated from
Co!!ea arabica L against S. aureus and E. coli
at the )34 of range 50H+00 1g2mL and 400H(00
1g2mL respecti#elyI howe#er, it did not show
acti#ity in the tested concentrations for C.
albicans. Ceports of =u&>ang et al.,
30
showed
)34 in the range of +./5H+0 mg2mL for C.
albicans. Lu et al.,
+4
found that ergosterol
deri#ati#es of Colletotrichum sp. isolated from
Artemisia annua inhibited S. aureus and B.
subtilis and Pseudomonas sp. at the range of
/5H!5 1g2mL and for C. albicans 50H+00
1g2mL. Endophytic Fusarium sp. from plant
Selaginella pallescens collected in the
7uanacaste conser#ation area of 4osta Cica
also shows potent acti#ity against Candida
albicans in agar diffusion assay
/3
. .ccording to
=u&>ang et al.,
30
most of the tested fungi
isolated from N. oleander possessed better
antibacterial and antifungal acti#ities than the
host plant. 3n the present study, antimicrobial
acti#ity of crude e"tracts of F. semitectum and
A. alternata, at +00 1g2mL concentration, were
not comparable statistically to the 6one
produced by the tetracycline $/0 1g2mL',
whereas, C. gloeosporioides is #ery near to the
standard. The acti#ity of )ycelia sterilia sp.+ at
+00 1g2mL concentration was statistically
significant to the positi#e control against S.
aureus. The crude e"tract of Fusarium
semitectum at +00 1g2mL was in par with the
positi#e control against B. cereus. Fusarium
semitectum and C. gloeosporioides were
inacti#e against P. aeruginosa. Bowe#er, the
6one of inhibition produced by A. alternata and
)ycelia sterilia sp.+ $/01g2mL' was statistically
significant with that of positi#e control against
the same pathogen. Fusarium semitectum did
not inhibit E. coli e#en at+00 1g2mL. Bowe#er,
the antibacterial acti#ity of A. alternata and C.
gloeosporioides against E. coli was higher than
that of positi#e control. 9oth the F. semitectum
and A. alternata showed no inhibition against S.
typhi, but the crude e"tract at ,0 1g2mL
concentration of )ycelia sterilia sp.+ and C.
gloeosporioides e"hibited higher 6one than the
positi#e control. None of inhibition of )ycelia
sterilia sp.+ against C. albicans at (01g2mL was
higher than that of flucona6ole at /0 1g2mL.
$Table /'.

Table 1
Identification and Screening the antimicrobial activity of endophytic
fungi isolated from #erium oleander

Endophytic
fungi iameter !one of inhibition in mm
4ode
Endophytic fungus
identified S. aureus B. cereus P. aeruginosa E.coli S. typhi C. albicans
*l.
%o
+ %os &+ Curvularia sp.


0
a



0
a



0
a



0
a
/O0
b



0
a

/ %os&/ Drechslera sp.


0
a



0
a



0
a



0
a



0
a



0
a

3 %os&3 Fusarium.sp.


0
a



0
a



0
a
4O0
c



0
a



0
a

4 %os&4 Cladosporium sp.+


0
a



0
a



0
a



0
a



0
a



0
a

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B - 688
5 %os&5 Cladosporium sp./


0
a



0
a



0
a



0
a



0
a



0
a

, %os&, )ycelia sterilia sp.+ 4O+
e
5.33O0.5(
e
(O+
b
(.33O0.5(
e
!O0
d
5.,!O0.5!
b

! %os&! Pn identified


0
a



0
a



0
a



0
a



0
a



0
a

( %os&( Pn identified


0
a



0
a



0
a



0
a



0
a



0
a

%os& hielavia terricola


0
a



0
a



0
a



0
a



0
a



0
a

+0 %os&+0 )ycelia sterilia sp./


0
a



0
a



0
a



0
a



0
a



0
a

++ %os&++ )ycelia sterilia sp.3


0
a



0
a



0
a



0
a



0
a



0
a

+/ %of&+ Aspergillus sp.

0
a


0
a


0
a


0
a


0
a


0
a

+3 %of&/ Cochliobolus sp.


0
a
4O0
d



0
a
5.33O0.5(
d
/O0
b



0
a

+4 %of&3
Fusarium
semitectum 3.,!O0.5(
de
+3.33O+.53
f



0
a



0
a



0
a



0
a

+5 %of&4
Curvularia
brachyspora /O0
c
/O0
b




0
a




0
a




0
a




0
a

+, %of&5
Alternaria
brassicola


0
a



0
a



0
a



0
a



0
a



0
a

+! %of&, Chaetomium sp. +O0
b



0
a



0
a



0
a
/.,!O0.5(
c
0
a

+( %of&! Alternaria alternata 3.33O0.5(
d
3.,!O0.5(
d
O0
c
3O0
b



0
a



0
a

+ %of&(


Colletotrichum
gloeosporioides +0.33O0.5(
f
4O0
d
0
a
4O0
c
,.,!O+.53
d
0
a

/0 %of& Cochliobolus sp.

0
a


0
a


0
a


0
a


0
a


0
a

/+ %of&+0
Cylindrocephalum
sp. 0
a
3O0
c



0
a



0
a



0
a



0
a

// %of&++ Colletotrichum sp.


0
a



0
a



0
a



0
a



0
a



0
a

/3 %of&+/ )ycelia sterilia sp.4


0
a



0
a



0
a



0
a



0
a



0
a

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B - 68'
/4 %of&+3 orula sp.


0
a



0
a



0
a



0
a



0
a



0
a

/5 %of&+4 Pn identified


0
a



0
a



0
a



0
a



0
a



0
a

/, %of&+5 Penicillium *p.


0
a



0
a



0
a



0
a



0
a



0
a

/! %ol&+

)ycelia sterilia sp.5


0
a



0
a



0
a



0
a



0
a



0
a

/( %ol&/ Bipolaris *p.


0
a



0
a



0
a



0
a



0
a



0
a

#os$ #erium oleander stem host isolate% #of$ #erium oleander flower host isolate% #ol$ #erium oleander leaf host isolate&
In each column% mean values followed by the same letter are not significantly different according to '()* at p + ,&,-(SPSS
ver 11&-)&

Table "
Antimicrobial activity of ethylacetate by agar well diffusion method

Endophytic
fungi
#oncentration
of e$tract
%g&mL
Antimicrobial activity 'iameter of inhibition !one in mm(

S& aureus .& cereus P&aeruginosa /& coli S& typhi 0& albicans

%of3 /0
40
,0
(0
+00
3.,!O0.5(
a
,.33O0.5(
c
(.33O0.5(
d
+/.,!O+.+5
f
+5.0O/.,4
g
+0.33O/.(
d
+/.,!O+.+5
e
+,.33O0.5(
f
+.,!O/.5+
hi
/+.00O+.00
i
0
a
0
a
0
a
0
a
0
a
0
a
0
a
0
a
0
a
0
a

0
a
0
a
0
a
0
a
0
a

0
a
0
a
0
a
0
a
0
a

%of! /0
40
,0
(0
+00
4.00O+.00
a
4.33O+.+5
ab
(.33O0.5(
d
++.,!O0.5(
ef
+5.33O0.5(
g
4.,!O0.5(
ab
(.00O0.00
c
+0.33O+.+5
d
+4.33O0.5(
e
+(.33O0.5(
h
5.,!O0.5(
b
(.33O0.5(
c
++.,!O0.5(
de
+/.33O+.+5
e
/+.00O+.!3
h

+.,!O0.5(
b
4.33O0.5(
cd
5.,!O0.5(
d
(.33O0.5(
e
+3.0O0.00
gh
0
a
0
a
0
a
0
a
0
a

0
a
0
a
0
a
0
a
0
a

%of( /0
40
,0
(0
+00
,.,!O+.+5
cd
+0.,!O0.5(
e
+5.33O0.5(
g
+,.33O+.5/
gh
//.0O0.00
-
3.30O0.5(
a
4.,!O0.5(
ab
5.,!O0.5(
b
(.,!O0.5(
cd
+3.0O+.00
e
0
a
0
a
0
a
0
a
0
a

/.00O0.00
b
3.00O0.00
bc
5.3O0.5(
d
(.33O0.5(
e
+/.,!O/.0(
gh
5.00O0.00
c
+4.0O+.00
e
+!.33O+.+5
g
/0.33O0.5(
h
/4.0O+.00
i
0
a
0
a
0
a
0
a
0
a

%os, /0
40
,0
(0
+00
,.00O+.00
bc
+0.,!O+.+5
e
+4.,!O+.+5
g
+(.0O/.0
h
/0.0O0.00
i
4.00O0.00
ab
(.,!O+.+5
cd
+3.0O+.00
e
+,.,!O+.+5
fg
+.33O+.+5
hi
5.,!O0.5(
b
+0.,!O+.+5
d
+/.33O0.5(
e
+3.,!O0.5(
f
+5.33O0.5(
g
+0.,!O+.+5
f
+,.0O0.00
i
/0.,!O+.+5
-
/4.,!O+.+5
;
/(.,!O+.+5
l
3.,!O0.5(
b
+/.,!O+.+5
d
/+.33O+.+5
h
/4.,!O+.+5
i
/!.33O+.+5
-
3.0O0.00
b
5.33O0.5(
c
!.,!O0.5(
d
+/.00O+.00
f
+4.,!O0.5(
g
Tetracycline /0 //.33O0.(
-
+0.33O0.5(
d
5.33O0.5(
b
++.33O+.+5
fg
+5.33O0.5(
f
&
:lucono6ole /0 & & & & & (.,!O0.5(
e
In each column% mean values followed by the same letter are not significantly different according to '()* at p + ,&,-&
1$2 #ot determined&
Int J Pharm Bio Sci 2013 Jan; 4(1): (B) 683 - 693


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B - 6'&


)igure 1.
/ndophytic fungi isolates of different parts of #erium oleander 3&



)igure ".
(icroscopic pictures of endophytic fungi with conidia from #erium oleander 3& a&
0olletotrichum sp&% b& 0haetomium sp&% c& 0urvularia sp&% d& *hielavia terricola% e& Alternaria
sp&% f& 0haetomium sp&% g& .ipolaris sp&% h& 0ylindrocephalum sp&
Int J Pharm Bio Sci 2013 Jan; 4(1): (B) 683 - 693


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B - 6'>


)igure *.
Antimicrobial activity of endophytic fungi by agar plugs method&




)igure +
Antimicrobial activity of ethyl acetate extract by agar well diffusion method

CONCLUSION

The present findings indicate that antimicrobial properties of endophytic fungi #ary with the
geographical distribution of the host plant. F. semitectum $%of&3', A. alternata $%of&!', C.
gloeosporioides $%of&(' and )ycelia sterilia sp.+ from Nerium oleander are potential sources of new
antibiotic and further research is re8uired to determine the chemical nature and efficacy of the
compounds.



Int J Pharm Bio Sci 2013 Jan; 4(1): (B) 683 - 693


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B - 6'*
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