Biol Fertil Soils (2002) 35:231–237
DOI 10.1007/s00374-002-0462-8
O R I G I N A L PA P E R
H. N. Asghar · Z. A. Zahir · M. Arshad · A. Khaliq
Relationship between in vitro production of auxins by rhizobacteria
and their growth-promoting activities in Brassica juncea L.
Received: 30 July 2001 / Published online: 13 April 2002
© Springer-Verlag 2002
Abstract Rhizobacteria were isolated from the rhizo-
sphere of different Brassica species and assayed for their
ability to produce auxins in vitro. The isolates varied
greatly in their potential for auxin production (ranging
from 0.33 to 11.40 µg ml–1). L-Tryptophan (an auxin pre-
cursor) addition to the media increased the auxin produc-
tion by several fold. Based upon in vitro auxin produc-
tion and growth promotion of B. juncea seedlings caused
by various isolates under gnotobiotic conditions, promis-
ing isolates were selected and tested in pot trial to ob-
serve their effects on growth, yield and oil content of the
same Brassica species. Results showed that seed inocu-
lation with different isolates of rhizobacteria significant-
ly increased plant height (up to 56.5%), stem diameter
(up to 11.0%), number of branches (up to 35.7%), num-
ber of pods per plant (up to 26.7%), 1,000-grain weight
(up to 33.9%), grain yield (up to 45.4%) and oil content
(up to 5.6%) over the uninoculated control. Isolate S54
gave the most promising and consistent results. Highly
significant correlations between L-TRP-derived auxin
production by plant growth-promoting rhizobacteria
(PGPR) in vitro and grain yield (r =0.77**), number
of pods (r =0.78**) and number of branches per plant
(r =0.77**) of B. juncea were found. It was hypothesized
that these PGPR may influence the growth and yield of
inoculated plants by production of auxins in the rhizo-
sphere of inoculated plants from the L-TRP present in the
root exudates, although other mechanisms of action
might have also contributed.
Keywords Rhizobacteria · Plant growth-promoting
rhizobacteria · Auxins · Brassica
H.N. Asghar · Z.A. Zahir · M. Arshad (✉) · A. Khaliq
Department of Soil Science, University of Agriculture,
Faisalabad-38040, Pakistan
e-mail: bio@fsd.paknet.com.pk
Tel.: +92-41-627793
Introduction
Beneficial rhizobacteria are often referred to as plant
growth-promoting rhizobacteria (PGPR). These encom-
pass all bacteria that inhabit plant roots and exert a posi-
tive effect by various mechanisms, ranging from a direct
influence (e.g. increased solubilization and uptake of nu-
trients or production of plant growth regulators), to an
indirect effect (e.g. suppression of pathogens by produc-
ing siderophores or antibiotics; Kloepper et al. 1989;
Glick et al. 1994). These beneficial bacteria are also re-
ferred to as yield-increasing bacteria (YIB; Chen et al.
1994).
Plant scientists became interested in the study of
PGPR due to their potential for improving crop growth
and yield. Direct growth promotion by PGPR was first
reported by Lifshitz et al. (1987). They studied the
growth promotion activity of Pseudomonas putida strain
GR 12–2 in gnotobiotic growth pouch assay and reported
that inoculation of rapeseed with GR12–2 significantly
increased root length, shoot height and phosphorus up-
take compared with an uninoculated control. In China,
results of multi-year trials on rapeseed indicated that ap-
plication of PGPR increased the yield by up to 11.5%
(Mei 1989). In a repeated field trial at four locations,
rapeseed inoculation with Bacillus cereus strain 83–10
increased the grain yield by 17.2% compared with an un-
inoculated control (Xia et al. 1990). Inoculation with
PGPR was also associated with increased oil content.
Kloepper et al. (1988) collected 4,000 bacterial strains
from the root zone of rapeseed and screened them at
4–14°C on the basis of growth promotion, metabolism of
seed exudates and root colonization. They further tested
887 strains for growth promotion of rapeseed in green-
house trials and reported that 35 strains increased grain
yield up to 57% compared to an uninoculated control in
one of the two test years. Three strains increased yield
by 6–13% during the 2-year trial. In another study,
Kloepper et al. (1991) found that inoculation with PGPR
increased the emergence and dry weight of 2-day-old
rapeseed seedlings by 123% and 79%, respectively, over
232
an uninoculated control. Similarly, a significant increase
in emergence rate was also observed due to inoculation
with certain root colonizing bacteria, including Pseudo-
monas (Kloepper et al. 1986). Bertrand et al. (2001) iso-
lated 13 rhizobacteria from rhizoplane and rhizosphere
of canola (Brassica napus) to evaluate their plant
growth-promoting activity. Eight of these 13 bacteria in-
duced a significant increase in root dry weight ranging
from 11–52%. The most important direct PGPR effect
was found with isolates belonging to the genus Phyllo-
bacterium (Bertrand et al. 2001). Chen et al. (1994) iso-
lated PGPR strains from the roots and rhizosphere soil
and used these to inoculate sterile peat. For many years
(1979–1990) they grew rapeseed inoculated with five
strains and observed a significant increase in yield
(11.5% over the uninoculated control). Significant yield
increases in many other crops such as wheat, maize and
potato have also been reported in response to inoculation
with PGPR (Iswandi et al. 1987; Javed et al. 1996;
Khalid et al. 1997; Zahir et al. 1998).
Plant growth-regulating substances are naturally oc-
curring organic compounds that influence physiological
processes in plants. Several studies have demonstrated
the potential of rhizosphere microflora to synthesize
plant growth regulators (PGRs) in vitro (Frankenberger
and Arshad 1995). Microflora that are able to produce
PGRs in vitro are present in appreciable numbers in
the rhizosphere of plants. Sarwar and Kremer (1995)
screened 16 different bacterial isolates belonging to the
genera Enterobacter, Xanthomonas, Pseudomnas, Alcali-
genes and Agrobacterium originating from the rhizo-
sphere of various plants. They found that all of the rhizo-
sphere isolates were more efficient producers of auxins
than soil isolates not associated with plant roots. Similar-
ly, 13 species of Azotobacter out of 14 (93%) strains iso-
lated from the roots of berseem, cotton, maize, pea
and wheat produced auxins (Apte and Shende 1981).
Fuentes-Ramirez et al. (1993) reported that all 18 strains
of the plant growth-promoting rhizobacterium, Aceto-
bacter diazotrophicus from 13 cultivars of sugarcane had
the ability to produce indole acetic acid (IAA) ranging
from 0.14 to 2.42 µg IAA ml–1. Arshad and Frankenberg-
er (1998) and Zahir et al. (2000) reported that addition of
tryptophan (TRP) as an auxin precursor substantially in-
creased auxin production.
Keeping this in mind, indigenous rhizobacteria were
isolated from the rhizosphere of different Brassica spe-
cies at different locations, and a relationship was deter-
mined between in vitro auxin production and their ef-
fects on various growth parameters in B. juncea.
Materials and methods
Isolation
Rhizosphere soil samples were collected from canola (B. napus
L.), toria (B. campestris L. var. Toria), Gobi sarsoon (B. carinata
A.Br.), sarsoon (B. campestris L.) and raya (B. juncea L. Czern &
Coss) from different locations. One hundred rhizobacterial cul-
tures were isolated by dilution plate technique using glucose pep-
tone agar medium (GPAM; Wollum II 1982). Bacterial colonies
showing prolific growth were selected and purified by further
streaking on fresh plates. Slants were prepared on GPAM and re-
frigerated at 4°C for further studies. These isolates were main-
tained by transferring them to new slants after every 20–30 days.
Auxin production
Sterilized broth of GPAM (25 ml) was put into glass tubes and in-
oculated with 0.1 ml rhizobacterial cultures (108–109 cells ml–1)
incubated at 28±2°C for 24 h with occasional shaking. The con-
tents of the tubes were filtered through Whatman filter paper No.2
before measuring auxin production as indole acetic acid (IAA)
equivalents. In measuring the IAA equivalents, 3 ml of the filtrate
were pipetted into test tubes and 2 ml Salkowski reagent
(2 ml 0.5 M FeCl3 +98 ml 35% HClO4) were added to it. The
tubes containing the mixture were left for 30 min for color devel-
opment. Intensity of the color was measured spectrophotometrical-
ly (detection limits 0.2–45 µg/ml) at 535 nm. Similarly, color was
also developed in standard solutions of IAA and a standard curve
was established by measuring the intensity of this color (Sarwar et
al. 1992). The same procedure, except for amendment of broth
with 5 ml 0.5% filter sterilized L-TRP solution, was repeated for
L-TRP-dependent auxin production by the rhizobacteria. Auxin
production by the two most efficient isolates (S89 and S54)
was also confirmed by HPLC-UV (Shimadzu SPD-10A) analysis
(column, CLC-ODS C18-Shim-pack, 4.6 mm internal diameter,
250 mm long; flow rate, 1.0 ml min–1, mobile phase, methanol and
water, 70:30; sample injected, 20 µl; detection, 280 nm).
Jar experiment
Inoculum of each culture was prepared (50 ml portions of the
GPAM broth) by incubation at 28±2°C for 4 days. Two sterilized
filter paper sheets were soaked in inoculum (108–109 CFU ml–1);
these filter papers were saturated with inoculum and then six sur-
face-sterilized seeds (with 0.1% acidified HgCl2 solution) of raya
(B. juncea L.) were placed on one sheet of the moist filter paper and
were covered with the other moist sheet so that they were sand-
wiched between the sheets. These sheets were then rolled and
placed in a glass jar. In the case of the uninoculated control, steril-
ized (autoclaved) inoculum was used. Sterilized Hoagland solution
(Hoagland and Arnon 1950) was applied to the jars to provide nutri-
ents. The jars were arranged according to a completely randomized
design with six replicates in a growth room maintained at 20°C. The
jars were kept in the dark during the pre-germination period. After
germination, 10 h of light were supplied daily. After 10 days, root
length, shoot length and fresh shoot weight were recorded.
Total root length
For the measurement of root length, fresh roots were cut into 1-cm
pieces and randomly spread in a tray, marked with gridlines, of
known area (A) in cm2 and total length of straight lines (H) in cm.
The total number of intersections (N) of roots with lines were re-
corded with a telecounter. Total root length (R) in cm was calcu-
lated as described by Newman (1966).
Data from ten promising isolates (labeled as PGPR) were subject-
ed to statistical analysis and these isolates were selected for pot
trial in a wirehouse.
Pot trial
The inocula for the pot trial were prepared by growing the selected
PGPR in nutrient broth at 28±2°C. The inoculum suspension
 233
Table 1 Auxin production by
rhizobacteria isolated from dif- Species Isolate IAA equivalents µg ml–1
ferent Brassica species (aver-
age of three replicates ± stan- Without L-TRP With L-TRP
dard error; IAA indole acetic Mean ± SE Mean ± SE
acid, L-TRP L-Tryptophan)
B. napus S1-S9 0.36±0.114 to 9.20±0.566 11.47±0.628 to 15.27±0.144
B. campestris var. Toria S10-SS27 1.66±0.675 to 10.92±1.436 2.42±0.75 to 19.13±0.412
B. carinata S28-S45 2.37±0.098 to 11.07±0.891 5.53±0.233 to 21.43±0.931
B. campestris S46-S73 0.33±0.18 to 5.95±0.024 4.93±0.943 to 24.43±0.223
B. juncea S74-S100 0.35±0.265 to 11.40±0.262 5.17±0.476 to 24.60±0.330

Table 2 Effect of rhizobacteria on shoot weight, root length and shoot length of a Brassica juncea seedlings in laboratory experiment
(average of six replicates ± standard error)

Isolate Shoot weight (mg) ± SE Root length (cm) ± SE Shoot length (cm) ± SE

S1-S100 6.67±3.043 to 45.0±3.909 1.03±0.423 to 18.8±0.664 0.95±0.397 to 8.4±0.285

(108–109 CFU ml–1) was injected into sterile peat (100 ml kg–1,
seed to peat ratio 1:1) and incubated for 24 h at 28±2°C before us-
ing it for seed coating. For inoculation treatments, seed dressing
was done with inoculated peat mixed with 10% sugar solution,
while for the uninoculated controls, the seeds were coated with the
same but sterilized (autoclaved) inoculum suspension. Six inoculat-
ed seeds of B. juncea were sown in pots having 12 kg pot–1 sandy
clay loam soil (pH 7.1; ECe, 3.5 dS m–1; organic matter, 1.1%; to-
tal N, 0.06%; available P, 20.6 mg kg–1 and extractable K, 300 mg
kg–1 soil). There were six replications and pots were arranged ran-
domly according to a completely randomized design in a wirehouse
under ambient light and temperature conditions. One week after
germination, the plants were thinned to one plant per pot. The fer-
tilizer NPK was applied at 50/75/50 kg ha–1, respectively. Pots
were irrigated when needed. Different yield and yield-contributing
parameters were recorded at maturity. The seed oil contents were
determined by using Soxhlets apparatus as described by the Asso-
Fig. 1 HPLC chromatogram of L-Tryptophan-dependent auxins
ciation of Official Analytical Chemists (1984). Data were analyzed
produced by isolate S89
statistically as described by Steel and Torrie (1980).

Results
Auxin production

The results (Table 1) revealed that auxins (expressed as

IAA equivalent in µg ml–1) were detected in liquid cul-
tures of bacterial isolates from all the Brassica species and
these increased when supplemented with L-TRP. Without
added L-TRP, the auxin production was highest (11.40 µg
ml–1) in the case of isolate S87 isolated from raya whereas
the isolate (S73) isolated from the rhizosphere of sarsoon
produced the lowest amount (0.33 µg ml–1). When L-TRP
was added to the medium, the highest auxin production
(24.6 µg ml–1 IAA equivalents) was by the isolate S89
from raya species while the minimum (2.42 µg ml–1) was
by a rhizobacterial strain S27 isolated from toria. The in- Fig. 2 HPLC chromatogram of L-Tryptophan-dependent auxins
crease in auxin production in response to L-TRP applica- produced by isolate S54
tion varied (ranging from 2.42 to 24.6 µg ml–1) with rhizo-
bacterial isolates. Auxin production by the most efficient Jar experiment
isolates, S89 and S54, was also confirmed by HPLC-UV
analysis. Study revealed that both isolates had the ability Data (Table 2) revealed that out of the 100 rhizobacterial
to produce IAA and indole-3-acetamide (IAM) from the isolates 68% increased the shoot weight, 30% decreased
added L-TRP (Figs. 1 and 2). it, while 2% of isolates had no effect on shoot weight in
234
Table 3 Effect of selected
rhizobacteria on shoot weight, Isolate Shoot weight (mg) Root length (cm) Shoot length (cm)
root length and shoot length of
Brassica juncea seedlings in Control 21.7 b 7.4 f 5.97 e
laboratory experiment (average S26 36.7 a 16.4 abc 7.6 abc
of six replicates). Means shar- S33 33.3 a 14.9 bcd 6.4 de
ing the same letter(s) do not S38 30.0 a 14.3 cd 6.6 de
differ significantly according to S54 41.7 a 13.5 d 8.4 a
Duncans Multiple Range Test S55 31.7 a 16.7 ab 7.1 bcd
(P =0.05) S74 45.0 a 17.8 a 7.9 ab
S82 40.0 a 18.8 a 8.3 a
S84 30.0 a 11.9 e 5.9 e
S87 40.0 a 18.0 a 6.8 cde
S89 35.0 a 18.7 a 7.8 abc

Table 4 Effect of selected rhizobacteria on the plant height, stem replicates). Means sharing the same letter(s) do not differ signifi-
diameter, no. of branches per plant, no. of pods per plant and cantly according to Duncans Multiple Range Test (P =0.05)
1,000-grain weight of Brassica juncea in pot trial (average of six

Isolate Plant height Stem diameter No. of branches No. of pods 1,000-grain Grain yield Oil contents
(cm) (mm) per plant per plant weight (g) (g/plant) (%)

Control 107.0 f 8.25 d 7.0 c 330.8 e 2.8 b 9.5d 41.9 c

S26 133.3 de 10.0 abc 9.5 a 409.8 abc 3.6 a 13.1ab 43.0 bc
S33 146.2 bcd 9.8 abc 8.0 bc 345.0 de 3.3 a 10.1 cd 42.1 c
S38 145.3 bcd 10.8 ab 8.5 ab 365.8 cde 3.3 a 11.3 bcd 42.0 c
S54 167.5 a 10.5 abc 8.5 ab 419.0 a 3.8 a 13.8 a 44.2 a
S55 126.0 f 9.3 cd 9.0 ab 403.0 abc 3.8 a 12.4 ab 42.4 c
S74 144.5 bcd 9.8 abc 8.0 bc 378.8 abcd 3.5 a 11.6 bc 42.4 c
S82 129.8 de 9.5 bcd 8.0 bc 370.8 bcde 3.6 a 11.1 bcd 42.9 bc
S84 158.8 ab 11.0 a 8.3 b 414.3 ab 3.6 a 13.1 ab 43.6 ab
S87 135.8cde 10.0 abc 9.0 ab 370.5 bcde 3.5 a 11.2 bcd 42.3 c
S89 150.0 bc 9.5 bcd 9.5 a 408.8 abc 3.5 a 13.1 ab 42.2 c

comparison with the uninoculated control. In the case of
root length, 73% of rhizobacterial isolates had positive
and 27% had negative effects. Shoot length was promot-
ed by 55% of rhizobacteria while 45% depressed shoot
length compared with the uninoculated control. Statistical
analysis (Table 3) of the results for 10 promising isolates
(out of 100 rhizobacteria) showed that shoot weight, root
length and shoot length of B. juncea seedlings were
significantly increased by inoculating with different iso-
lates. Isolate S74 produced the maximum shoot weight
(45 mg), 107% higher than the uninoculated control.
The increase in plant root length was highest (154%
over the uninoculated control) when inoculated with S82
isolate. S84 was the least effective isolate but still it pro-
duced 60% longer roots as compared to the uninoculated
control. The maximum shoot length of 8.4 cm, that was
43% longer than the uninoculated control, was observed
when seeds of B. juncea were inoculated with bacterial
isolate S54. The effect of S33, S38, S84 or S87 on shoot
length was not statistically different from the uninoculat-
ed control.
Pot trial
Statistical analysis of data (Table 4) revealed that some
of the PGPR isolates had a significant effect on different
parameters of B. juncea in the pot trial. None of the pa-
rameters had significant correlation with in vitro auxin
production by the rhizobacterial isolates without L-TRP.
However, when incubated in the presence of L-TRP, their
auxin production was significantly correlated with some
of the parameters observed.
Maximum plant height (167.5 cm) was obtained upon
inoculation with isolate S54, which was 57% higher than
the uninoculated control. Minimum increase in plant
height was observed with S55, which was 17.8% higher
than the uninoculated control, but this increase was sta-
tistically non-significant.
Inoculation with various PGPR increased (15.2% to
33.3%) stem diameter compared with the uninoculated
control (Table 4). Isolate S84 was the most effective in
increasing stem diameter significantly (33.3% over the
uninoculated control). It was also statistically better than
isolates S82, S55 and S89. The smallest stem diameter
(9.3 mm) was recorded in plants where the seed was in-
oculated with S55, which was not statistically different
from the uninoculated control.
The majority of the PGPR significantly increased the
number of branches per plant (Table 4) except for S82,
S74 and S33. Plants inoculated with S26 and S89 had the
same number of branches and these were 36% higher
than the uninoculated control. The number of branches
per plant correlated significantly (r =0.77 at P =0.01)
with in vitro auxin production by PGPR isolates in the
presence of L-TRP.

Six out of 10 isolates had a significant effect on the
number of pods per plant. The maximum number of pods
per plant (419) was produced in response to inoculation
with S54 and the minimum with S33. The increase in the
number of pods per plant as a result of inoculation
ranged from 4 to 26.7% compared with the uninoculated
control. A significant correlation (r =0.78 at P =0.01)
was observed between the number of pods per plant and
in vitro auxin production by PGPR when incubated in
the presence of L-TRP.
Inoculation with all the PGPR increased 1,000-grain
weight significantly (Table 4). The increase varied from
17.1% to 33.9% compared with the uninoculated control.
The maximum 1,000-grain weight (3.8 g) produced by
inoculation with isolate S54 and S55 was 33.9% higher
than the uninoculated control. All treatments had approx-
imately the same effect on 1,000-grain weight, but dif-
ference among the treatments was statistically non-sig-
nificant.
Table 4 reveals that most of the isolates (S54, S26,
S84, S89, S55 and S74) increased grain yield significant-
ly over the uninoculated control. The other isolates (S38,
S87, S82, and S33) were non-significant on grain yield.
Maximum grain yield (13.8 g/plant) was obtained when
inoculated with isolate S54 being 45% higher than the
uninoculated control. The grain yield had a significant
correlation (r =0.77 at P =0.01) with in vitro auxin pro-
duction by PGPR when incubated in the presence of
L-TRP.
Isolates S54 and S84 significantly increased the oil
content of raya compared with the uninoculated control.
Isolate S54 produced the highest oil contents (up to
44.2%), which were 6% greater than the uninoculated
control. Isolate S38 was the least effective in increasing
oil content of raya.
In pot trial, the plant height ranged from 107.0 to
167.5 cm, stem diameter from 8.2 to 11.0 mm, number
of branches from 7.0 to 9.5, number of pods per plant
from 330.8 to 419.0, 1,000-grain weight from 2.8 to
3.8 g, grain yield from 9.5 to 13.8 g/plant and oil con-
tents from 41.9% to 44.2% in response to inoculation
with selective isolates.
Discussion
In this study all rhizobacterial isolates produced auxins
(expressed as IAA equivalents) in the presence and ab-
sence of L-TRP although much variation was observed in
the potential to produce auxins. Auxin production by all
isolates increased when culture medium was supple-
mented with an auxin precursor, L-TRP. The L-TRP-
derived auxins were increased up to 24.6 µg ml–1which
was 184-fold more than that without L-TRP. This in-
crease in auxin production due to L-TRP also varied with
rhizobacterial isolates. This is in agreement with the
findings of many workers (Fuentes-Ramirez et al. 1993;
Leinhos and Vacek 1994; Sarwar and Kremer 1995;
Zahir et al. 2000). Leinhos and Vacek (1994) reported
235
auxin production by Pseudomonas and Acinotobacter
isolated from wheat and rye rhizosphere ranging from
0.01 to 3.98 mg IAA-equivalents per liter culture medi-
um. Similarly, Sarwar and Kremer (1995) screened 16
different bacterial isolates belonging to the genera
Enterobacter, Xanthomonas, Pseudomonas, Alcaligenes
and Agrobacterium collected from the rhizosphere of
various plants for auxin production. Four isolates of
Pseudomonas spp. from bulk soil were also capable of
producing auxins in vitro. Likewise, Fuentes-Ramirez et
al. (1993) reported that all 18 strains of Acetobacter
diazotrophicus isolated from the rhizosphere of 13 culti-
vars of sugarcane had the ability to produce IAA in vitro
ranging from 0.14 to 2.42 µg IAA ml–1.
In agreement to our results, various other workers
have also reported stimulation of auxin production in
response to L-TRP application (Sarwar et al. 1992;
Martens and Frankenberger 1993; Zahir et al. 2000).
Mordukhora et al. (1991) screened 216 strains of the ge-
nus Pseudomonas for their ability to produce IAA and
observed that Pseudomonas was stimulated to synthesize
IAA upon the addition of tryptophan to the medium.
Similar to our findings, auxin production by ten Azoto-
bacter cultures isolated from maize rhizosphere ranged
from 1.5 to 3.1 mg l–1. Selected Azotobacter cultures
were monitored for substrate- (L-TRP-) dependent auxin
production and it was revealed that supplementation with
L-TRP (10–3 M) resulted in a 171% increase in auxin
production by the Azotobacter culture Z4 (Zahir et al.
2000).
Similar to our findings regarding variation in the abil-
ity to biosynthesize auxin among rhizobacteria, many
other workers also have reported that auxin production
by microbial isolates varied greatly within species and/or
strains of the same species (Fuentes-Ramirez et al. 1993;
Mansour et al. 1994; Zahir et al. 2000).
In our laboratory trial, results showed significant in-
crease in shoot length (43%), root length (154%) and
shoot weight (107%) in response to inoculation with
rhizobacterial isolates. Kloepper et al. (1991) also report-
ed similar results. They found that inoculation with
PGPR increased the emergence and dry weight of rape-
seed seedlings while Noel et al. (1996) found that inocu-
lation with PGPR significantly promoted the early seed-
ling and root growth of canola. The results of Glick et al.
(1997) also support the findings of our study. They ex-
amined the early development of canola seedling with
P. putida GR12-2 and found that this bacterium promot-
ed root and shoot growth. Similarly Bertrand et al.
(2001) reported a significant increase in root dry weight
ranging from 11% to 52% in canola upon inoculation
with PGPR.
In the pot trial, seed inoculation with PGPR signifi-
cantly increased plant height, stem diameter, number of
branches, 1,000-grain weight, number of pods per plant,
grain yield and oil content from 5.6% to 56.5% over the
uninoculated control. In agreement to our results, field
trials conducted by Mei (1989) and Chen et al. (1994) re-
vealed that rapeseed inoculation with Bacillus cereus in-
236
creased the grain yield by 11.5%. Likewise, replicated
field trials were conducted over 2 years at two locations
and up to 11.4% and 14.7% increases in yields was ob-
served with B. cereus strains A-47 over control yields
each year. In addition to yield promotion, PGPR inocula-
tion also enhanced rate of seedlings tiller formation and
plant dry weight (Xia et al. 1990). De Freitas et al.
(1997) reported that PGPR significantly increased plant
height, number and weight of pods and seed yield of ca-
nola in potted soil.
The observed growth promotion and increase in crop
yields in response to PGPR inoculation could be ex-
plained on the basis of various mechanisms. Most PGPR
strains may have multiple mechanisms of action, which
accounts for the observed beneficial effects on plant
growth. Many researchers are of the view that a very im-
portant mechanism of direct growth promotion may be
the production of plant growth regulators by PGPR
(Lifshitz et al. 1987; Frankenberger and Arshad 1995).
Several studies have frequently demonstrated the ability
of various PGPR to produce plant growth regulators
(PGRs) in vitro. In this study, we have made an attempt
to find a correlation, if any, between auxin production by
PGPR in vitro and growth promotion caused by them. It
was found that L-TRP-derived auxin production by PGPR
in vitro was significantly correlated with grain yield
(r =0.77), number of branches (r =0.77) and number of
pods per plant (r =0.78). In vitro auxin production by
PGPR in the absence of L-TRP was not significantly corre-
lated with the growth parameters. It is highly likely that
plant roots might have released L-TRP in the root exudates
resulting in more auxin production in the rhizosphere. This
increased auxin production in vivo (rhizosphere) might
have improved the growth and yield of inoculated plants.
This study reveals that among the PGPR isolates, the
S54 isolate is relatively more consistent in improving the
different growth parameters of B. juncea, which may
imply that it could be used as PGPR for enhancing the
yield and oil contents of B. juncea in the field. More work
is in progress to test this isolate in agronomic studies.
References
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