Abstract Evaluation of some endophytes have been carried for their possible antimicrobial activity from various parts of medicinal plants belonging to Jalgaon Maharashtra (India).A total of seventy eight bacterial endophytes and one hundred fourty two fungal endophytes were isolated from the aerial and underground parts of selected medicinal plants .Fifteen positive endophytic bacterial isolates and fourteen positive endophytic fungal isolates possess antifungal and antibacterial activity respectively. Bacterial isolates KB4 from roots of roots of P.glabra, NB6 from stem of E.globulus and HB3 from rhizomes of C.longa have strong antifungal activity. Endophytic fungi AFR1, AFR4, AFR7 from roots of A.vera possess strong antibacterial activity against S. typhi in dual culture assay

R E Jalgaonwala

R T Mahajan

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R E Jalgaonwala et al. / International Journal on Pharmaceutical and Biomedical Research (IJPBR) 
Vol. 1(5), 2010, 136-141 
 
Evaluation of Endophytes for their 
Antimicrobial activity from Indigenous 
Medicinal Plants belonging to North 
Maharashtra region India . 
R E Jalgaonwala1*     ,   B V Mohite2 and R T Mahajan3 
1, 2 Lecturer Department of Biotechnology 
3 Reader and Head Department of Biotechnology 
Moolji Jaitha College Jalgaon, 425001.M.S. India. 
Abstract 
Evaluation of some endophytes have been carried for their possible antimicrobial activity from various parts of 
medicinal plants belonging to Jalgaon Maharashtra (India).A total of seventy eight  bacterial endophytes and one 
hundred fourty  two fungal  endophytes were isolated from  the aerial and underground parts of selected  medicinal 
plants .Fifteen positive endophytic bacterial isolates and fourteen positive endophytic fungal isolates possess 
antifungal and antibacterial  activity respectively. Bacterial isolates KB4 from roots of roots of P.glabra, NB6 
from stem of E.globulus and HB3 from rhizomes of C.longa have strong antifungal activity. Endophytic fungi 
AFR1, AFR4, AFR7 from roots of A.vera possess strong antibacterial activity against S. typhi in dual culture 
assay. 
Keywords: Antibacterial activity, Antifungal activity, Endophytes, Medicinal plants. 
Introduction    
Endophytes are microbes that inhabits plant tissues  in their life cycle without causing any apparent harm to their 
host [1].Their presence implied a symbiotic interaction, in all the plants investigated until now[2] .Endophytes were 
mentioned for first time by Bray in 19th century [3]. Endophytes have been found in nearly all plant families [4]. 
Dryfuss and Chapera [5] estimated that there may be at least one million species of endophytic fungi alone. 
Recently endophytes are viewed as outstanding source of secondary metabolites and bioactive antimicrobial natural 
products. Endophytic fungi are of biotechnological interest  due to their  potential use as genetic vectors , 
metabolites [6,7,8] and biological control agents[9,10,11,12].Present study was carried out by selecting indigenous 
medicinal plants growing in Jalgaon viz. as Azadirachta indica A Juss , Curcuma longa L., Eucalyptus globulus 
Dehnh ,Musa paradiasica L., Pongamia glabra Vent, Aloe vera L. Morrayo konengi L. Sprengel ., Osimum 
sanctum L.,Mant .  Endophytic microorganisms from indigenous medicinal plants were evaluated for their 
antimicrobial activities.     
Materials and Methods 
Mature healthy plant materials were collected by sampling different parts  of the selected medicinal plants, fifteen 
samples were taken from each tree five each from roots , inner bark and leaves, From each sample ten sub samples 
were separated further for isolation of endophytes. Samples were  immediately brought to laboratory and were used 
within 24 hrs .Samples were washed in running tap water for 10 minutes to remove , soil particles and adhered 
debris , and finally washed with distilled water. Surface sterilization was done using methodology described by 
Petrini 1992[13] With some modifications .Samples were immersed in 70% ethanol 1-3 minutes and 4% aqueous 
solution of sodium hypochlorite 3 minutes again 1 minutes with 70% ethanol 1 minutes and 0.1% mercury chloride 
2 minutes .Finally rinse with sterile distilled water three times. Samples tissues were selected by aseptic cutting 
using sterile knife and inner tissues were excised. Tissues were inoculated on to the water agar and Tryptic soy agar 
media for two to six days at 25o C to isolate endophytic bacteria and for isolation of endophytic fungi inoculated on 
to the potato dextrose agar containing Streptomycin as to avoid bacterial growth and incubation allowed for 6-25 
days at 25o C. 
ISSN : 0976- 285X 136
R E Jalgaonwala et al. / International Journal on Pharmaceutical and Biomedical Research (IJPBR) 
Vol. 1(5), 2010, 136-141 
 
Antifungal activity by Bacterial Endophytes 
Antifungal activity was screened using dual culture method in which both endophyte and test fungi were inoculated 
in same media plate. Five days old culture disks (5mm diameter)of test fungi were inoculated  at the centre of 
potato dextrose agar plate and endophytic bacteria were spot inoculated at three corner of the plate and  incubated 
for four to eight  days at 270  C. Antifungal activity was indicative as mycelia growth of test fungus prohibited in 
the direction of active endophyte, the level of inhibition was calculated by subtracting the distance (mm)of fungal 
growth in the direction of an antagonist colony from the endophyte growth  radius. The width of inhibition zone 
between the pathogen and endophyte was evaluated as inhibition zone and ranked as  .+,<2mm;++  >,2-10mm;+++ 
,10 > mm[14]. 
Antibacterial activity fungal Endophytes 
Antibacterial activity was screened using dual culture technique with some modification. Suspension of  24 hrs 
old culture of pathogenic bacteria was spread on sterile nutrient agar  plate on to which five day old disc (5mm 
diameter) of  endophytic fungi was kept  and incubated at 270c for 24 -48 hrs. Antibacterial activity was calculated 
by measuring zone of inhibition produce by endophytic fungi against pathogenic bacteria.  
Inhibition zone diameter index :+++> 10mm, ++5-10mm+<5mm –no zone [15] 
Table 1: Antifungal activity by some positive endophytic bacteria from indigenous medicinal plants 
Host Family Tissue Isolates Positive 
Isolates 
         Antifungal activity 
An       Aa        Tk          Fo           Pf     
A. indica Meliaceae Ls 09 (01)NMB1 ++ - ++ ++ +++ 
  Rt 07 (01)NMB4 - - +++ - - 
  St 06 (01)NMB6 ++ ++ ++ ++ ++ 
C.longa Zingiberaceae Rz 06 (01)HB3 +++ +++ +++ ++++ ++ 
E.globulus Myrtaceae Ls 07 (01)NB1 - - - - - 
  St 05 (01)NB6 +++ +++ +++ +++ +++ 
  Rt 04 (00) - - - - - 
M.paradiasica Musaceae Ls 10 (01)BB1 + + + ++ + 
  St 06 (02)BB2 ++ - - + + 
  Rt 04 (02)BB5 - - ++ - - 
P.glabra Leguminoseae Ls 08 (01)KB1 ++ - + + - 
  St 06 (01)KB2 - - - - - 
  Rt 06 (01)KB4 +++ +++ +++ +++ +++ 
 
a)An: Aspergillus niger, Aa: Aspergillus avamori, Tk: Trycoderma konningi, Fo: Fusarium oxysporium,  
b)   Pc: Penicillium fumicalsuri     
c) Antifungal activity:  +, < 2mm; ++, 2-10mm; +++, >10mm. 
ISSN : 0976- 285X 137
R E Jalgaonwala et al. / International Journal on Pharmaceutical and Biomedical Research (IJPBR) 
Vol. 1(5), 2010, 136-141 
 
 d) Ls: Leaves, St: Stem, Rt: Roots, Rz: Rhizomes. 
Table2: Antibacterial activity by some positive endophytic fungi from indigenous medicinal plants. 
Host Family Tissue Isolate Positive 
Isolates 
                    Antibacterial activity 
Ec   St       Bs        Sa          Pv       Pf 
A. vera                Liliaceae    Rts 20 (03)ALF1 
       ALF4 
       ALF7 
- ++
+ 
++
+ 
++
+ 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
C. longa       Zingiberaceae    Rz 08 (02)  HF4 
         HF6 
++ 
- 
- 
+ 
3.2 
++ 
++ 
+ 
++
+ 
++
+ 
++ 
- 
M. konengi    Rutaceae   Ls 20 (01) KTP1 - - ++ - + - 
  St 17 (01) KTP6 + + + - - - 
  Rt 15 (01) KTP8 + + + + + - 
M. paradiasica Musaceae Ls 07 (01) BBF5 + + - - + - 
  St 06 (00) - - - - - - 
  Rt 05 (00) - - - - - - 
O.sanctum    Labiatae Ls 20 (03)   TF3 
          TF9 
          TF5 
+ 
+ 
++ 
- 
+ 
++ 
- 
- 
+ 
+ 
+ 
+ 
+ 
- 
+ 
- 
- 
- 
  St 15 (01)   TF1 ++ + + - + - 
  Rt 09 (01) TF17 + + + + + - 
 
a)Inhibition zone diameter index :+++> 10mm, ++5-10mm+<5mm –no zone 
b) Ec: Escherichia coli, St: Salmonella typhi, Bs: Bacillus subtilis, Sa: Staphylococcus aureus ,  
Pv: Protease vulgaris     
 c) Ls: Leaves, St: Stem, Rt: Roots, Rz: Rhizomes.       
Results and Discussion 
 Plants have long provided mankind with a source of medicinal agents,with natural products once serving as source 
of all drugs[16].Recently biological controls or the uses of microorganisms or their secretions to prevent diseases 
offer an attractive alternative or supplement to disease management without the negative impact of chemical 
ISSN : 0976- 285X 138
R E Jalgaonwala et al. / International Journal on Pharmaceutical and Biomedical Research (IJPBR) 
Vol. 1(5), 2010, 136-141 
 
control[17] .A total of seventy eight bacterial endophytes and one hundred and fourty two fungal endophytes were 
obtained from roots ,stem and leaf tissues of seven different medicinal plants used by local population of Jalgaon. 
KTP1from roots of M. konengi, TF5 from leaves of O.sanctum were strong candidates 
Table 1 summarizes antifungal activity by bacterial endophytes .About fourteen positive Endophytic bacteria were 
screened for their antifungal activity against pathogenic fungi.  Isolate KB4 from roots of P.glabra, NB6 from stem 
of E.globulus and HB3 from rhizomes of C. longa are potent as per antifungal activity was concerned this is 
followed by isolates NMB1,NMB6, BB1 from A.indica and M.paradiasica, rest other endophytic bacteria are less 
effective. Highest antifungal activity was recorded in endophytic bacteria KB4.  
Table 2 shows antibacterial activity by fungal endophytes. fourteen positive endophytic fungi were obtained 
,endophytic fungi  HFR4 ,HFR6  from rhizomes of C. longa were  powerful isolates for antibacterial activity 
against pathogenic microbes .Fungal endophytes AFR1,AFR4,AFR from roots of A.vera  showed strong inhibition  
against S.typhi . Fungal endophyte KTP1 from roots of M. konengi, TF5 from leaves of O.sanctum found to have 
good antifungal activity. Other fungal endophytes were less active. Thus the activity spectra of endophytes were 
greatly different depending upon isolates, suggesting that several substances participated in antimicrobial activity. 
Antifungal activities of plant endophytic fungi have been reported by some researchers[18,19,20,21,22].Gram 
negative endophytic bacteria with potent antibacterial activity from roots of Solanum sp was reported by Long , 
2003[15]. Population density of endophytes seems to be highest in aerial tissues than the underground tissues. The 
diversity of endophytes obtained from healthy plant tissues suggests that an even broader flora of endophytes might 
be found across diverse plant species. These results suggest the presence of an active principle in the strains 
showing positive biological activities so needs to be enhancing production of the secondary metabolite of the 
interest. It is clear that endophytes isolated may be beneficial to their host .Antibacterial resistance especially 
among Gram negative bacteria is an important issue that has created a number of problems in treatment of 
infectious diseases and necessitates the search for alternative drug or natural antibacterial [22].This aspect of 
antimicrobial activity will be further investigated to enhance production of secondary metabolites of interest. 
Therefore, any information and research on endophyte plant symbiosis is of value. Recently biological controls or 
the uses of microorganisms or their secretions to prevent diseases offer an attractive alternative or supplement to 
disease management without the negative impact of chemical control [17].Plants have long provided mankind with 
a source of medicinal agents, with natural products once serving as source of all drugs [16]. Endophytic 
microorganisms are excellent sources of bioactive natural products that can be use to satisfy demand of 
Pharmaceutical and Medical Industries.   
ACKNOWLEDGMENT 
We are thankful to Principal M J College Jalgaon for providing all necessary facilities to complete the project. 
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a.                                                                                                   b.   
       Figure1.Endophytic fungi isolated from branch of Morrayo konengi (a) and roots of A.vera (b.) 
     
a.                                                                                 b. 
          Figure1. a. Antifungal activity by endophytic bacteria KB4 against Trycoderma konningi and 
                          b. antibacterial activity by endophytic fungi ALF1against S.typhi 
  
ISSN : 0976- 285X 140
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Vol. 1(5), 2010, 136-141 
 
 
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Evaluation of Endophytes for their Antimicrobial activity from Indigenous Medicinal Plants belonging to North Maharashtra region India

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Evaluation of Endophytes for their Antimicrobial activity from Indigenous Medicinal Plants belonging to North Maharashtra region India

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