ARTICLE IN PRESS

Microbiological Research 163 (2008) 173181

www.elsevier.de/micres

Screening of free-living rhizospheric bacteria for

their multiple plant growth promoting activities
Farah Ahmad, Iqbal Ahmad, M.S. Khan

Department of Agricultural Microbiology, Aligarh Muslim University, Aligarh 202002, India

Accepted 10 April 2006

KEYWORDS Summary
PGPR;
Plant growth promoting rhizobacteria (PGPR) are known to influence plant growth by
Indoleacetic acid;
various direct or indirect mechanisms. In search of efficient PGPR strains with
Siderophores and
multiple activities, a total of 72 bacterial isolates belonging to Azotobacter,
antifungal activity
fluorescent Pseudomonas, Mesorhizobium and Bacillus were isolated from different
rhizospheric soil and plant root nodules in the vicinity of Aligarh. These test isolates
were biochemically characterized. These isolates were screened in vitro for their
plant growth promoting traits like production of indoleacetic acid (IAA), ammonia
(NH3), hydrogen cyanide (HCN), siderophore, phosphate solubilization and antifungal
activity. More than 80% of the isolates of Azotobacter, fluorescent Pseudomonas and
Mesorhizobium ciceri produced IAA, whereas only 20% of Bacillus isolates was IAA
producer. Solubilization of phosphate was commonly detected in the isolates of
Bacillus (80%) followed by Azotobacter (74.47%), Pseudomonas (55.56%) and
Mesorhizobium (16.67%). All test isolates could produce ammonia but none of the
isolates hydrolyzed chitin. Siderophore production and antifungal activity of these
isolates except Mesorhizobium were exhibited by 1012.77% isolates. HCN
production was more common trait of Pseudomonas (88.89%) and Bacillus (50%).
On the basis of multiple plant growth promoting activities, eleven bacterial isolates
(seven Azotobacter, three Pseudomonas and one Bacillus) were evaluated for their
quantitative IAA production, and broad-spectrum (active against X three test fungi)
antifungal activity. Almost at all concentration of tryptophan (50500 mg/ml), IAA
production was highest in the Pseudomonas followed by Azotobacter and Bacillus
isolates. Azotobacter isolates (AZT3, AZT13, AZT23), Pseudomonas (Ps5) and Bacillus
(B1) showed broad-spectrum antifungal activity on Muller-Hinton medium against
Aspergillus, one or more species of Fusarium and Rhizoctonia bataticola. Further
evaluation of the isolates exhibiting multiple plant growth promoting (PGP) traits on
soilplant system is needed to uncover their efficacy as effective PGPR.
& 2006 Elsevier GmbH. All rights reserved.

Corresponding author. Tel.: +91 9412371170; fax: +91 571 2703516.

E-mail address: iqbalahmad8@yahoo.co.in (I. Ahmad).

0944-5013/$ - see front matter & 2006 Elsevier GmbH. All rights reserved.
doi:10.1016/j.micres.2006.04.001
 ARTICLE IN PRESS
174
Introduction
Plant growth promoting rhizobacteria (PGPR) are
a heterogeneous group of bacteria that can be
found in the rhizosphere, at root surfaces and in
association with roots, which can improve the
extent or quality of plant growth directly and or
indirectly. In last few decades a large array of
bacteria including species of Pseudomonas, Azos-
pirillum, Azotobacter, Klebsiella, Enterobacter,
Alcaligens, Arthobacter, Burkholderia, Bacillus
and Serratia have reported to enhance plant
growth (Kloepper et al., 1989; Okon and Laban-
dera-Gonzalez, 1994; Glick, 1995). The direct
promotion by PGPR entails either providing the
plant with a plant growth promoting substances
that is synthesized by the bacterium or facilitating
the uptake of certain plant nutrients from the
environment. The indirect promotion of plant
growth occurs when PGPR lessen or prevent the
deleterious effect of one or more phytopathogenic
micro-organisms.
The exact mechanisms by which PGPR promote
plant growth are not fully understood, but are
thought to include (i) the ability to produce or
change the concentration of plant growth regula-
tors like indoleacetic acid, gibberellic acid, cyto-
kinins and ethylene (Arshad and Frankenberger,
1993; Glick, 1995), (ii) asymbiotic N2 fixation
(Boddey and Dobereiner, 1995), (iii) antagonism
against phytopathogenic microorganisms by pro-
duction of siderophores (Scher and Baker, 1982),
antibiotics (Shanahan et al., 1992) and cyanide
(Flaishman et al., 1996), (iv) solubilization of
mineral phosphates and other nutrients (De Freitas
et al., 1997; Gaur, 1990). Most popular bacteria
studied and exploited as biocontrol agent includes
the species of fluorescent Pseudomonas and Bacil-
lus. Some PGPR may promote plant growth indir-
ectly by affecting symbiotic N2 fixation, nodulation
or nodule occupancy (Fuhrmann and Wollum,
1989). However, role of cyanide production is
contradictory as it may be associated with deleter-
ious as well as beneficial rhizobacteria (Bakker and
Schippers, 1987; Alstrom and Burns, 1989).
In addition to these traits, plant growth promot-
ing bacterial strains must be rhizospheric compe-
tent, able to survive and colonize in the
rhizospheric soil (Cattelan et al., 1999). Unfortu-
nately, the interaction between associative PGPR
and plants can be unstable. The good results
obtained in vitro cannot always be dependably
reproduced under field conditions (Chanway and
Holl, 1993; Zhender et al., 1999). The variability in
the performance of PGPR may be due to various
environmental factors that may affect their growth
F. Ahmad et al.
and exert their effect on the plant. The environ-
mental factors include climate, weather condi-
tions, soil characteristics or the composition or
activity of the indigenous microbial flora of the soil.
To achieve the maximum growth promoting inter-
action between PGPR and nursery seedlings it is
important to discover how the rhizobacteria exert-
ing their effects on plant and whether the effects
are altered by various environmental factors,
including the presence of other micro-organisms
(Bent et al., 2001).
Therefore, it is necessary to develop efficient
strains in field conditions. One possible approach is
to explore soil microbial diversity for PGPR having
combination of PGP activities and well adapted to
particular soil environment. So keeping in view the
above constrains, the present study was designed
to screen certain rhizospheric bacterial isolates
belonging to Azotobacter, Mesorhizobium ciceri,
fluorescent Pseudomonas and Bacillus for their
multiple plant growth promoting activities.
Materials and methods
Isolation and characterization
All the isolates of Azotobacter, Pseudomonas and
Bacillus were isolated from the rhizospheric soil of
different crops (mustard, barseem, wheat, sugar-
cane, brinjal, onion, cauliflower, cabbage and chick
pea) grown in vicinity of Aligarh, UP, India.
Mesorhizobium was isolated from the nodules of
chickpea on yeast extract mannitol agar containing
per liter of distilled water: 10 g mannitol, 0.5 g
K2HPO4, 0.2 g MgSO4  7H2O, 0.1 g NaCl, 1.0 g yeast
extract, 3.0 g CaCO3, 15 ml Congo red (1:400
aqueous solution), 20 g agar, pH 6.87.0. M. ciceri
isolates were confirmed by nodulation assay under
sterile pot condition as described by Vincent
(1970).
Whereas other bacteria were isolated on their
respective media, Azotobacter on Jensens medium
containing per liter of distilled water: 20 g sucrose,
1 g K2HPO4, 0.5 g MgSO4  7H2O, 0.5 g NaCl, 0.1 g
K2SO4, 0.005 g Na2MoO4, 20 g agar, pH 6.9, Pseudo-
monas on Kings B medium containing per liter of
distilled water: 10 g proteose peptone, 10 ml
glycerol, 1.5 g K2HPO4, 1.5 g MgSO4, 20 g agar, pH
7.2 and Bacillus on nutrient agar (NA) containing
per liter of distilled water: 5.0 g peptone, 1.5 g
yeast extract, 1.5 g beef extract, 5.0 g NaCl, 20 g
agar, pH 7.2. Bacterial cultures were maintained on
the respective slants. Fluorescence of Pseudomo-
nas colonies was observed on Kings B medium
 ARTICLE IN PRESS
Screening of free-living rhizospheric bacteria
under UV exposure. All the microbiological media
and media ingredients were purchased from Hi-
Media Lab. Pvt. Ltd., Mumbai, India.
The bacterial isolates were characterized by
their cultural conditions, morphological and bio-
chemical characteristics (hydrolysis of starch, lipid
and chitin, utilization of glucose, sucrose, lactose,
mannitol, citrate and catalase reactions) using
standard methods (Cappuccino and Sherman,
1992). Biotin prototrophy was determined by
growing the isolates on Bacto biotin assay medium
(Difco Laboratories) for 4872 h at 2872 1C.
In vitro screening of bacterial isolates for
their plant growth promoting (PGP) activities
Assay for indoleacetic acid (IAA) production
IAA production was detected by the modified
method as described by Brick et al. (1991).
Quantitative analysis of IAA was performed using
the method of Loper and Scroth (1986) at different
concentrations of tryptophan (0, 50, 150, 300, 400
and 500 mg/ml). Bacterial cultures were grown for
72 h (Azotobacter) and 48 h (Pseudomonas and
Bacillus) on their respective media at 2872 1C.
Fully grown cultures were centrifuged at 3000 rpm
for 30 min. The supernatant (2 ml) was mixed with
two drops of orthophosphoric acid and 4 ml of the
Salkowski reagent (50 ml, 35% of perchloric acid,
1 ml 0.5 M FeCl3 solution). Development of pink
colour indicates IAA production. Optical density
was taken at 530 nm with the help of spectro-
photometer Spectronic 20 D+. Concentration of IAA
produced by cultures was measured with the help
of standard graph of IAA (Hi-media) obtained in the
range of 10100 mg/ml.
NH3 production
Bacterial isolates were tested for the production
of ammonia in peptone water. Freshly grown
cultures were inoculated in 10 ml peptone water
in each tube and incubated for 4872 h at 2872 1C.
Nesslers reagent (0.5 ml) was added in each tube.
Development of brown to yellow colour was a
positive test for ammonia production (Cappuccino
and Sherman, 1992).
HCN production
All the isolates were screened for the production
of hydrogen cyanide by adapting the method of
Lorck (1948). Briefly, nutrient broth was amended
with 4.4 g glycine/l and bacteria were streaked on
modified agar plate. A Whatman filter paper no. 1
soaked in 2% sodium carbonate in 0.5% picric acid
solution was placed in the top of the plate. Plates
175
were sealed with parafilm and incubated at
2872 1C for 4 days. Development of orange to red
colour indicated HCN production.
Siderophore production
Bacterial isolates were assayed for siderophores
production on the Chrome azurol S agar medium
(Sigma, Ltd.) described by Schwyn and Neilands
(1987). Chrome azurol S agar plates were prepared
and divided into equal sectors and spot inoculated
with test organism (10 ml of 106 CFU/ml) and
incubated at 2872 1C for 4872 h. Development of
yelloworange halo around the growth was con-
sidered as positive for siderophore production.
Phosphate solubilization by test bacteria
All isolates were first screened on Pikovskayas
agar plates for phosphate solubilization as de-
scribed by Gaur (1990). Quantitative analysis of
solubilization of tricalcium phosphate in liquid
medium was made as described by King (1932).
Briefly, the test isolates were inoculated in 25 ml
Pikovskayas broth and incubated for 4 days at
2872 1C. The bacterial cultures were centrifuged
at 15,000 rpm for 30 min. Supernatant (1 ml) was
mixed with 10 ml of chloromolibidic acid and the
volume was made up to 45 ml with distilled water.
Cholorostannous acid (0.25 ml) was added and the
volume was made up to 50 ml with distilled water.
The absorbance of the developing blue colour was
read at 600 nm. The amount of soluble phosphorus
was detected from the standard curve of KH2PO4.
Antifungal assay
The agar well diffusion method as adopted
earlier (Mehmood et al., 1999) was used. The
bacterial isolates tested for their antifungal activ-
ity were fully grown in the respective broth media.
Test fungi were grown on Sabaroud dextrose agar
(SDA), (per liter of distilled water: 40 g dextrose,
10 g peptone, 20 g agar) slants. The spores were
scraped and suspended in 10 ml of sterile normal
saline solution (NSS). Diluted spore suspension
(0.1 ml, 105 CFU/ml) of the fungi was spread on
Muller Hinton agar (per liter of distilled water:
300 g beef infusion, 17.5 g casein acid hydrolysate,
1.5 g starch, 20 g agar, pH 7.2), NA and SDA plates.
Wells of 8 mm diameter were punched into the agar
medium and filled with 200 ml (2  107 CFU/ml) of
bacterial culture. Nutrient broth was taken as
negative control and 100 mg/ml antifungal antibio-
tic, nystatin was used as positive control. The
plates were incubated for 56 days at 2872 1C. The
antifungal activity was evaluated by measuring the
growth inhibition zone against test fungi.
 ARTICLE IN PRESS
176 F. Ahmad et al.

Results Pseudomonas (11.11%) and Bacillus (10%). All

isolates were negative for chitin hydrolysis whereas
Isolation and biochemical characterization positive for ammonia production.

On the basis of cultural, morphological and Quantitative assay of IAA production by

biochemical characteristics a total of 66 soil
isolates were grouped into Azotobacter, Bacillus,
fluorescent Pseudomonas as described in Bergeys
Manual of Determinative Bacteriology (Holt et al.,
1994). Six nodule isolates of chickpea (Cicer
aretinum) were characterized as M. ciceri (Nour
et al., 1994). General features of the test isolates
are illustrated in Table 1. All the isolates of M.
ciceri were prototrophic for biotin whereas 91.49%
isolates of Azotobacter, 83.33% isolates of fluor-
escent Pseudomonas and 80% isolates of Bacillus
were prototrophic for biotin.
Plant growth promoting traits of test isolates
Screening results of PGP traits are depicted in
Figs. 1 and 2. IAA production was shown in all the
isolates of fluorescent Pseudomonas followed by
Azotobacter (83.3%), M. ciceri (83.3%) and Bacillus
(20%). Phosphate solubilization was detected in 80%
of isolates of Bacillus followed by Azotobacter
(74.47%), fluorescent Pseudomonas (55.56%) and M.
ciceri (16.67%). Production of siderophore and
antifungal activity was simultaneously exhibited
by isolates of Azotobacter (16.22%), fluorescent
selected isolates
A total of 11 selected isolates of Azotobacter
(seven), fluorescent Pseudomonas (three) and
Bacillus (one) were tested for the quantitative
estimation of IAA in the presence of different
concentrations of tryptophan. With no addition of
tryptophan, production of IAA was not observed.
With the addition of tryptophan from 50 to 500 mg/
ml the production of IAA was increased. The
production of IAA was highest in isolates of
fluorescent Pseudomonas, followed by Azotobacter
and Bacillus, respectively. Amongst the Azotobac-
ter, isolates AZT26 produced highest amount of IAA
followed by AZT34AZT134AZT234AZT14AZT20 as
depicted in Table 2.
Antifungal activity of the test isolates
Antifungal activity of AZT1, AZT3, AZT9, AZT13,
AZT20, AZT23, Ps5 and B1 was checked against
Aspergillus sp., Fusarium solani, F. ciceri, F.
oxysporum, Rhizoctonia bataticola using three
different media, MH, NA and SDA (Table 3). The
antifungal activity of strains tested varied with

Table 1. Biochemical characterization of the test isolates

Biochemical characters Azotobacter species Mesorhizobium Fluorescent Bacillus species

ciceri Pseudomonas

Number of isolates 47 6 9 10
Pigmentation Transparent, milky, Fluorescent
some becomes blackish green
brown on aging
Colony morphology Watery, mucilaginous Pin head, Button shaped Serrated
shrink, serrated mucilaginous margins
margins white
Polysaccharide production + +  
Gram reaction, cell shape  rods  rods  rods + rods
Growth on N2 free medium +   
Catalase, citrate test 100 100 100 100
Hydrolysis
Starch 68.09 55.56 80
Lipid 48.94 50 77.78 80
Biotin prototrophy 91.49 100 83.33 80
Carbohydrate utilization
Glucose 63.83 83.33 55.56 80
Lactose 70.21 16.67 11.11 20
Sucrose 78.72 83.33 33.33 80
Mannitol 36.17 16.67 70
 ARTICLE IN PRESS
Screening of free-living rhizospheric bacteria 177

IAA production

Phosphate solubilization

NH3 production

100
90
80
Percent isolates

70
60
50
40
30
20
10
0
Azotobacter spp. Mesorhizobium Fluorescent Bacillus spp.
ciceri Pseudomonas

Figure 1. Direct PGP activities of test isolates.

Siderophore production
Antifungal activity
90
80 HCN production

70
Percent isolates

60
50
40
30
20
10
0
Azotobacter Mesorhizobium Fluorescent Bacillus spp.
spp. ciceri Pseudomonas

Figure 2. Indirect PGP activities of test isolates.

Table 2. Production of indoleacetic acid by selected bacterial isolates grown on their respective medium

Isolate designation IAA production (mg/ml7SD) at different tryptophan concentrations (mg/ml)

0 50 150 300 400 500

AZT1 ND 1.4770.25 3.5370.32 6.5770.31 9.3770.15 11.8370.21

AZT3 ND 1.7770.21 5.0070.20 8.9070.30 11.2770.12 13.4770.45
AZT13 ND 1.6070.30 3.7770.25 7.0770.15 9.7370.25 12.8070.20
AZT9 ND 1.2770.31 3.5370.25 6.6370.40 8.5770.25 10.1770.25
AZT20 ND 1.2770.25 3.4070.20 6.4070.20 8.3370.25 10.4770.15
AZT23 ND 1.5070.20 3.6070.30 6.4770.31 8.9370.25 11.9070.20
AZT26 ND 2.1370.15 4.6770.15 7.6770.15 10.4370.47 15.0070.26
Ps5 ND 3.0070.20 7.4770.25 13.3770.35 16.8070.20 18.7070.30
Ps7 ND 2.6070.20 5.9070.20 11.3070.36 15.1770.25 18.0770.25
Ps9 ND 3.6070.20 6.1070.20 11.4770.15 14.9370.15 22.0270.20
B1 ND ND ND 3.4070.20 5.0370.15 7.0370.15

NDnot detectable.
 For Azotobacter Jensens, Pseudomonas Kings B, Bacillus nutrient media was used.
 178
Table 3. Antifungal activity of the test isolates on different media

Test isolates Media used Zone size (mm 8 SD)

Aspergillus sp. Fusarium solani Fusarium ciceri Fusarium oxysporum Rhizoctonia bataticola

AZT1 MH 22.6770.58 ND ND 15.6770.58 ND

NA 18.3370.58 ND ND 14.3370.58 ND
SDA 13.0071.41 ND ND 16.0071.00 ND
AZT3 MH 30.5070.50 23.5370.50 17.6770.58 30.8370.76 15.0071.00
NA 25.6770.58 18.6771.15 15.3370.58 22.3370.58 ND
SDA 19.0071.00 25.8370.76 21.3371.15 24.1770.29 ND
AZT9 MH 25.0071.00 ND ND 13.3371.15 ND

ARTICLE IN PRESS
NA 21.0071.00 ND ND ND ND
SDA 16.6770.58 ND ND ND ND
AZT13 MH 31.5070.50 21.0071.00 16.6770.58 17.6770.58 16.5070.50
NA 26.3370.58 16.00 14.5070.71 16.00 13.8370.76
SDA 22.0072.00 17.00 21.0071.00 17.5070.50 ND
AZT20 MH 19.0071.00 ND ND 15.6770.58 ND
NA 16.00 ND ND ND ND
SDA ND ND ND ND ND
AZT23 MH 17.5070.50 16.3370.58 ND 18.6770.58 ND
NA 14.00 ND ND 15.3370.58 ND
SDA ND ND ND 19.0071.00 ND
Ps5 MH 15.5070.50 15.3370.58 15.3370.58 19.0071.00 ND
NA 13.8370.7 12.0071.00 12.0071.00 14.8370.76 ND
SDA ND ND ND 12.0071.00 ND
B1 MH 15.5070.50 16.5070.50 16.5070.50 20.1770.76 ND
NA 12.8370.29 13.3370.58 13.3370.58 15.5070.50 ND
SDA 11.5070.71 14.2570.35 14.2570.35 12.6770.58 ND

F. Ahmad et al.
NDnot detected.
 ARTICLE IN PRESS
Screening of free-living rhizospheric bacteria
inhibition zones in diameter from 11.50 to
35.00 mm. Strains AZT3 and AZT13 induced larger
inhibition zones compared to the other strains.
AZT1 showed activity against Aspergillus sp. and F.
oxysporum on all three media. AZT3 and AZT13
showed good antifungal activity against all the
fungi but no activity was detected on SDA and or NA
against R. bataticola. AZT9 and AZT20 isolate
showed activity against Aspergillus sp. on all three
media whereas antifungal activity against F. oxy-
sporum was observed on MH medium. AZT23 was
effective against Aspergillus sp., F. oxysporum and
F. solani but results varied with different media.
PS5 and B1 could also exhibit broad-spectrum
activities against test fungi. MH medium was most
appropriate medium for screening of antifungal
activity and Aspergillus sp. was most susceptible
organism (Table 3).
Discussion
Plant rhizosphere is known to be preferred
ecological niche for various types of soil micro-
organisms due to rich nutrient availability. It has
been assumed that inoculation with diazotrophic
bacteria like Rhizobium, Azotobacter and Azospir-
illum enhanced the plant growth as a result of their
ability to fix nitrogen. However, despite of exten-
sive research efforts only rhizobia have been shown
to increase yields from dinitrogen fixation. Growth
promotion may be attributed to other mechanisms
such as production of plant growth promoting
hormones in the rhizosphere and other PGP
activities (Arshad and Frankenberger, 1993; Glick,
1995). In the present investigation 47 isolates of
Azotobacter and 25 isolates belonging to M. ciceri,
fluorescent Pseudomonas and Bacillus species were
screened in vitro for PGP activities. IAA production
was detected in all the test isolates of fluorescent
Pseudomonas, 83.3%, of both Azotobacter and M.
ciceri isolates. Our findings of IAA production in
Azotobacter isolates are in agreement with other
workers (Gonzalez-Lopez et al., 1986; Jagnow,
1987; Nieto and Frankenberger, 1989).
Phosphate solubilization was most frequently
encountered by Bacillus isolates (80%), followed
by Azotobacter, Pseudomonas and least by Mesor-
hizobium isolates. However, production of ammo-
nia was a common trait in all selected group of
bacteria. Siderophore production was detected
among some isolates of Azotobacter (12.77%),
followed by Pseudomonas and Bacillus isolates.
However, 50% and 80% isolates of Bacillus and
Pseudomonas were detected positive for HCN
179
production. While antifungal activity was shown
by 12.77% of Azotobacter isolates, followed by
Pseudomonas (11.11%) and Bacillus isolates (10%).
Some of the above-tested isolates could exhibit
more than two or three PGP traits, which may
promote plant growth directly or indirectly or
synergistically. Similar to our findings of multiple
PGP activities among PGPR have been reported by
some other workers while such findings on indigen-
ous isolates of India are less commonly explored
(Gupta et al., 1998). On the basis of preliminary
screening, quantitative analysis of IAA production
was made on seven Azotobacter isolates, three
fluorescent Pseudomonas and one Bacillus isolate.
There was an increase in the level of IAA with the
increasing concentration of tryptophan (50500 mg/
ml). Similar trend of IAA production with the
increasing concentration of tryptophan was also
reported by Barazani and Friedman (2000). Isolates
AZT26, AZT3, AZT13 could produce relatively high
concentration of IAA compared to other Azotobac-
ter isolates. Production of high levels of IAA by
fluorescent Pseudomonas is a general characteristic
of our test isolates. Similar high level of IAA
production was recorded by other workers (Xie et
al., 1996). The production of IAA was found
dependant upon bacterial isolates and concentra-
tion of tryptophan. Such findings may have direct
practical application, although intrinsic ability of
bacteria to produce IAA in the rhizosphere depends
on the availability of precursors and uptake of
microbial IAA by plant (Arshad and Frankenberger,
1993).
Another important trait of PGPR, that may
indirectly influence the plant growth, is the
production of siderophores. They bind to the
available form of iron Fe3+ in the rhizosphere, thus
making it unavailable to the phytopathogens and
protecting the plant health. In the present inves-
tigation six isolates of Azotobacter and the fluor-
escent Pseudomonas strain Ps5 showed multiple
PGP activities including siderophore production and
antifungal activities against one or more test fungi.
On the basis of data obtained it could be stated
that: (i) sensitivity of test fungi was in order of
Aspergillus sp.4F. oxysporum4F. solani4Rhizoc-
tonia bataticola; (ii) MullerHinton medium was
best out of the three tested media to detect the in
vitro antifungal activity which is probably due to
the non-interfering composition of this medium
with the assay system; and (iii) isolate AZT3 and
AZT13 demonstrated broad spectrum antifungal
activity against the five tested fungi. The anti-
fungal activity of the test isolates indicated a close
relationship between production of HCN and side-
rophores. The antifungal activity of the test
 ARTICLE IN PRESS
180
isolates might be due to the production of side-
rophore and HCN or synergistic interaction of these
two or with other metabolites. However, role of
chitinase was not expected as these isolates were
negative for chitin hydrolysis. Several studies have
demonstrated that production of siderophores,
other secondary metabolites and lytic enzymes by
Pseudomonas strains was most effective in control-
ling the plant root pathogens including F. oxyspor-
um and R. solani (OSullivan and OGara, 1992;
Nagrajkumar et al., 2004). Further studies on the
performance of these isolates and their mutants on
the growth of plant will uncover the mechanism
and potential of these PGPR exhibiting multiple
PGP traits.
Acknowledgements
We thank the Chairman, Department of Agricul-
tural Microbiology for providing necessary facilities
for this work, and Mr. Farrukh Aqil for technical
help.
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