Agricultural Science Research Journals Vol. 2(5); pp.

240-249, May 2012

Available online http://www.resjournals.com/arj
ISSN-L:2026-6073 International Research Journals

Full Length Research Paper

Effect of pantoea agglomerans on growth promotion

and yield of rice
1

KHALIMI K , *SUPRAPTA DN and NITTA Y

1

Laboratory of Biopesticide, Faculty of Agriculture Udayana University, Jl. PB. Sudirman, Denpasar Bali Indonesia
2
Laboratory of Crop Science, College of Agriculture, Ibaraki University, Ami, Ibaraki Japan.
*Corresponding authors email: biop@dps.centrin.net.id, Tel. /Fax: +62-361-255346.
ABSTRACT

Effect of Pantoea agglomerans isolated from the rhizosphere of groundnut was tested for the ability to
promote the growth and increase the yield of rice cultivar Cicih Medang Selem, a local cultivar of Bali.
Two isolates of P. agglomerans were used in a green house experiment, namely PaJ and BS2a.
Treatments with P. agglomerans were done by soaking the rice seeds with bacterial suspension before
sowing and application of compost formula at transplanting time. Some growth and yield parameters
were observed such as fresh and dry weights of shoot, fresh and dry weights of root, length of stem,
diameter of stem, leaf area, chlorophyll content, macro nutrients content in the leaf, number of tillers,
number of panicles per hill, percentage of pithy grain per panicles, dry weights of 1,000 grains and
weight of grain per hill. Results of this study showed that the treatment with P. agglomerans, as single
isolate (PaJ or BS2a) or in a mixture (PaJ and BS2a) significantly increased the plant growth and yield
of rice. All the plant growth and yield parameters of treated plants were significantly higher than those
of un-treated control plants (P<0.05). However, there was no significant difference on the growth and
yield parameters among treatments with P. agglomerans, either treatments with single strain or two
strains in a mixture. In general, treatment with P. agglomerans in a mixture resulted in better rice
performance than those of single strain. These results suggested that, mixture of application of both
the strains of P. agglomerans could be well utilized to promote the growth and to increase the yield of
rice.
Key Words: plant growth promoting rhizobacteria, Pantoea agglomerans, rice growth, rice yield.

INTRODUCTION
Rice (Oryza sativa L.) is one of the staple foods of more
than 60% of the worlds population (Umashankari and
Sekar, 2011). In Indonesia, the dependency of
Indonesian people to rice as their main food is 92-95%
(Machmur, 2010). The average consumption rate of rice
per capita per year of Indonesian people is 139 kg. This
amount is relatively higher than other countries in the
world (Dwijosumono, 2011). The average rice yield of
Indonesia was 4.56 tones/ha which was relatively lower
than other rice growing countries, such as Australia, 8.22
tones/ha; Japan, 5.85 tones/ha and China 6.06 tones/ha

(USDA, 2004). These differences demonstrated a

possibility to enhance the rice productivity in Indonesia by
improving the plant growth, including application of the
plant growth promoting rhizobacteria (PGPR) (Kloepper
et al., 1989; Glick, 1995).
The PGPR are bacteria that can be found in the
rhizospehere, a thin layer of soil immediately surrounding
the plant roots, and exert a positive effect on the plants. A
large array of bacteria including species of
Pseudomonas, Enterobacter, Azospirillum, Azotobacter,
Burkholderia, Bacillus and Serratia have been reported

KHALIMI et al.

as plant growth promoting rhizobacteria (PGPR) to

enhance the plant growth (Kloepper et al., 1989; Glick,
1995).
The mechanisms by which the PGPR promote the plant
growth has not been understood clearly, however,
several researchers reported that the mechanisms of
PGPR in promoting the plant growth are through the
increase of water and nutrients uptake, nitrogen fixation,
production of phytohormones, production of ACC
deaminase, phosphate solubization, production of
antibiotics which can suppress the growth of plant
pathogens, systemic induced resistance (Wei et al.,
1996; Thakuria et al., 2004).
Several researches have been reported the use of
rhizobacteria on plant growth promotion and yield. The
level of growth promotion and yield increment varied
according to the species of rhizobacteria and plants
species. Seed treatment of rice with Pseudomonas
fluorescens strain Pf1 in powder formulation at the rate
10 g/kg seeds significantly increased the yield in
comparison with control (Vidhyasekaran et al., 1997).
Biswas et al. (2000) reported that the rice plant
inoculated with Rhizobium sp. IRBG74 isolated from
Sesbania cannabina Linn. & Merrill could increase the
yield by 11.6%, while Alam et al. (2001) proved that the
treatment of rice root with the suspension of Azotobacter
nigricans, A. armeniacus, Bacillus sphaericus, B.
megaterium, Enterobacter and Xanthobacter for 24 h
could increase the yield by 15.03%. The increase of yield
of corn by 85% was obtained by Hameeda et al. (2008)
when the corn seeds were soaked in the suspension of
Serratia marcescens EB 67. Treatment of corn seed with
7
the suspension of Azotobacter at 10 cfu/g increased the
yield of corn by 19.76% (Sharifi et al., 2011).
One of the rhizobacteria that potentially promote the
plant growth and increase the yield of plant is Pantoea
agglomerans (formerly Enterobacter agglomerans).
Dursun et al. (2010) reported that the foliar spray of the
8
suspension of P. agglomerans isolate FF (10 cfu/ml)
increased the yield of tomato and cucumber by 43.87%
and 23.32%, respectively. There is no report available on
the use of P. agglomerans on plant growth promotion and
yield of rice Bali local variety. The current study has been
conducted to evaluate the effectiveness of P.
agglomerans as a plant growth promoter in Cicih Medang
Selem, one of Balis rice cultivar.

MATERIALS AND METHODS

Pantoea agglomerans strains
Two strains of Pantoea agglomerans, namely PaJ and

241

BS2a isolated from the root of groundnut were obtained

from the collection of the Laboratory of Biopesticide,
Faculty of Agriculture Udayana University. These strains
have been tested for their ability to promote the growth of
soybean, corn and tomato under green house condition.
In order to rapidly identify the inoculants, lyvofroxacin
resistant derivatives of both strains were selected
according to the following procedure. The bacterial
strains were grown overnight in Luria medium (10 g
Tryptone, 10 g Natrium Chloride, 5 g Yeast Extract, and
water to make 1 liter) at 280 C in a rotary shaker at 100
rpm. After centrifugation at 5,000 rpm for 5 minutes, the
pellet of each strain was washed two times by
suspending in 10 ml of sterile 0.1 M citrate buffer (pH
5.0). Then, the pellet was suspended in 100 ml of citrate
buffer containing 0.5 mg/ml levofloxacin and incubated
for 2 hours in a rotary shaker at 280C. Following the
incubation, the cells were pelleted down and washed
twice with 0.1 M phosphate buffer (pH 6.7) and the pellet
was suspended in 100 ml of phosphate buffer and serially
diluted up to 10-4 in phosphate buffer. Finally, 100 l
-3
-4
each of dilutions of 10 and 10 were plated into Luria
Agar plates containing 0.5 mg/ml levofloxacin and
incubated at 28 C. Individual colonies appeared in the
plates were used for this study.

Seedling Preparation
The rice seeds (cultivar Cicih Medang Selem) were
soaked in sterile distilled water containing 1% NaOCl
(sodium hypochlorite) for 5 minutes and rinsed with
sterile distilled water (five times). The seeds were then
soaked for 30 minutes in the bacterial suspension of P.
agglomerans strain Paj and BS2a individually as well as
with mixture of both the strains at the concentration of
8
10 cfu/ml. The non-treated seeds (control) were soaked
in sterile distilled water. The rice seeds were then
germinated and grown in a seedling tray filled with sterile
fertile soil for two weeks. These rice seedlings were
transplanted into plastic bucket filled with 5 kg soil per
bucket.
Formulation Development
The P. agglomerans strains PaJ and BS2a were
formulated in compost formulation. Three compost
formulations were developed in this study, namely 1) Pj
containing strain PaJ, 2) Bs containing strain BS2a and
3) PB containing the mixture of strains PaJ and BS2a.
8
Bacterial suspension (10 ml/kg) at 1.9 x 10 cfu/ml was
mixed with 45 day-old compost containing rice straw,
fresh leaf of rain tree (Samanea saman), rice bran and
sucrose (70:20:10:2, w/w/w/w). This mixture was put in a

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Agric. Sci. Res. J

plastic box and incubated for a week in the dark at room

o
temperature (28 + 2 C).

(DMRT) at 5% level of significance using SAS software

version 6.12 (SAS Institute, Gary, NC, USA).

Green House Experiment

Measurement of Chlorophyll and Nutrients Content

Treatment with P. agglomerans in compost formula was

done in a green house. This experiment was designed in
randomized block design (RBD) with four treatments
namely Pj, Bs, PB and C (control). Each treatment was
replicated six times, thus there were 24 experimental
units in this experiment. Each experiment unit consisted
of 10 rice plants. The application of P. agglomerans
compost formula was done once at transplanting time by
mixing 50 g compost with soil in each bucket. Rice
seedlings without treatment of P. agglomerans (C) were
planted in buckets filled with 50 g sterile compost.

Measurements of total chlorophyll content and nutrients

content were done at 44 DAS. Total chlorophyll content
(SPAD unit) was determined with a chlorophyll-meter
SPAD-502 (Konica Minolta, Japan). For nitrogen and
phosphorus analysis, dried leaf samples were digested
with sulphuric acid and hydrogen peroxide. Nitrogen
content was determined by Kjeldhals method and
phosphorus
content
was
determined
by
spectrophotometer after mixing the sample with Barton
reagents (Nadeem et al., 2006). Concentrations of
Potassium were determined using a Gallenkamp Flame
Photometer at 767 nm (Oyewale, 2005).

Colonization
Rhizosphere

of

Pantoea

agglomerans

in

the

Colonization of P. agglomerans in the root of rice plants

was evaluated 30 days after sowing (DAS). Sample of 1
g of root was ground in a mortal with 9 ml of 0.1 M
phosphate buffer (pH 6.7). Aliquot of 1 ml from each
-6
suspension was serially diluted up to 10 in distilled
-3
-6
water and 1 ml each of dilutions of 10 and 10 were
plated into Luria Agar plates containing 0.5 mg/ml
o
levofloxacin. The culture plates were incubated at 30 C
for 24 h. The number of colonies appeared was
calculated as each colony forming unit (cfu).

Plant growth and yield analysis

The samples for analysis of growth parameters were
taken at 30, 44, 58 days after sowing (DAS) to determine
the final dry weight (leaves, stem, roots, and total), plant
height, and leaf area. The net assimilation rate (NAR) in
grams per square centimeter of leaf surface and per day
was calculated as follows: NAR = (W2-W1) (logL2-logLl) /
(L2-Ll) (to-tl), where W1, and W2 are total dry weight and
L1, and L2 are total leaf area at times t1 and t2,
respectively. The relative growth rate (RGR) in grams per
day was calculated according to Alvim (1960) as follow:
RGR = (logW2-logW1) / (t2 t1), where W2 and W1 are
dry mass at time t2 and t1.
Measurement of yield components such as number of
panicles per hill, number of grains per panicle,
percentage of pithy grains per panicle, dry weight of
1,000 grains, and weight of grains per hill was done at 88
DAS. All data were subjected to the Analysis of variance
(ANOVA) followed by the Duncans multiple range test

RESULTS AND DISCUSSION

Rhizosphere
inoculants

colonization

of

P.

agglomerans

The P. agglomerans strains PaJ and BS2a were

originally isolated from rhizosphere of peanut plants.
However, they successfully colonized the roots of rice cv.
Cicih Medang Selem. This is confirmed through reisolation of P. agglomerans from the root of treated rice
plants. The number of colonies of P. agglomerans in the
roots of rice treated with PB (the mixture of isolates PaJ
6
and BS2a) was 2.56+0.67 x 10 cfu/g of roots, which was
slightly higher than those of rice plants treated with either
6
6
Pj or Bs as single isolate (2.42 X 10 and 2.47 X 10 ,
respectively), however no significant difference (P>0.05)
was detected in the three inoculation treatments.
Meanwhile, no lyvofroxacin resistant P. agglomerans was
found in the root of rice plants of control (Table 1). These
colonization rates were similar to that of PGPR reported
previously by Reding et al. (1991) in which the
colonization rates of Xhantobacter sp. strain JW-KR1 in
rice roots varied from 105 to 107 cfu/g of root.

Plant growth
In general treatment with P. agglomerans could promote
and improve the growth of rice plants. The values of net
assimilation rate (NAR) and relative growth rate (RGR) of
rice treated with P. agglomerans as single isolate (Pj and
Bs) or in the mixture (PB) were obviously higher than
those of rice without P. agglomerans treatment. Rice

KHALIMI et al.

243

Table 1. Population density of Pantoea agglomerans in the roots of rice plants

30 days after sowing
No.
1
2
3
4

Treatments

Population density of P. agglomerans

6
( 10 cfu/g of roots)
0 a*
2.41 b
2.47 b
2.56 b

C (control)
Pj
Bs
PB

* Values in the same columns followed by the same letters are not significantly different
(P>0.05) according to the Duncans Multiple Range Test (DMRT).
** Values in parenthesis
( ) indicating
percentage
compared
to controlbut
(%).the
No significant
differencethewas
detected ofinincreases
the threewhen
inoculation
treatments,
colonization was significantly greater in the treatments than that in control plants.

0.4

RGR
PB

0.2

Bs

0
30 DAS 44 DAS 58 DAS

Pj

gr/day

gr/cm2/day

NAR
0.4

PB

0.2

Bs

0
30 DAS 44 DAS 58 DAS

Pj

Fig. 1: Graphs of NAR (left) and RGR (right) of rice plants at different ages with and without treatment of P. agglomerans. DAS: days
FigurePB:
1: treatment
Graphs ofwith
NAR
andof RGR
of rice
plants
at different
treatment
ofPaJ.
P.
after sowing.
the (left)
mixture
strains(right)
PaJ and
BS2a,
Bs: treatment
withages
strainwith
PaJ,and
Pj: without
treatment
with strain
agglomerans. DAS: days after sowing. PB: treatment with the mixture of strains PaJ and BS2a, Bs: treatment with
strain PaJ, Pj: treatment with strain PaJ.

Table 2. Content of macro nutrients and chlorophyll in the leaf of rice treated with Pantoea agglomerans
Treatment
C
Pj
Bs
PB

Nitrogen (%)
4.4 a*
7.0 b
(59.09)**
7.2 b
(63.67)
7.6 b
(72.72)

Phosphorus (%)
2.8 a
4.4 b
(57.14)
5.0 b
(78.57)
4.6 b
(64.28)

Potassium (%)
2.7 a
3.6 b
(33.33)
4.4 b
(62.96)
3.8 b
(40.74)

Content of chlorophyl (SPAD unit)

31.02 a
38.25 b
(23.30)
39.12 b
(26.11)
39.32 b
(26.76)

*Values in the same columns followed by the same letters are not significantly different (P>0.05) according to
the Duncans Multiple Range Test (DMRT).
** Values in parenthesis ( ) indicating the percentage of increases when compared to control (%).

treated with PB (mixture of two isolates, PaJ and BS2a)

showed the highest NAR and RGR at all observations
(30, 44 and 58 DAS), while the lowest was shown by rice
of control (Figure 1). The highest NAR and RGR occurred
at 44 DAS at all treatments. Rice treated with PB showed
the NAR at 0.028 g/cm2/day, which was slightly higher
than those of rice treated with Pj or Bs, but obviously
higher than that of control. Similar trend was also
observed on RGR, in which rice treated with PB showed

the RGR at 0.034 g/day which was higher than other

treatments.
The increase in NAR and RGR values was in line with
the increase of the content of macro nutrients and
chlorophyll in the leaf. Treatment with P. agglomerans
significantly (P<0.05) increased the content of macro
nutrients such as nitrogen, phosphorus and potassium,
and increased the content of chlorophyll (Table 2).
However, there is no significant difference was detected

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Agric. Sci. Res. J

Table 3. Fresh and dry weights of shoots and roots, number of maximum tillers, length of Stem, and diameter of stem of
rice cv. Cicih Medang Selem treated with Pantoea agglomerans.
Treatments

Fresh
weight of
shoot (g)

Dry
weight of
shoot (g)

Fresh
weight of
root (g)

Dry
weight of
root (g)

Lenght of
Stem (cm)

Diameter of
stem (cm)

Leaf area
2
(cm )

120.40 a *
175.54 b
(45.79)**
229.06 b
(90.25)
252.97 b
(110.12)

51.07 a
96.40 b
(88.76)
93.50 b
(83.08)
118.62 b
(132.26)

83.97 a

11.04 a

63.94 a

0.59 a

60.05 a

201.28 b
(139.70)
236.24 b
(181.34)
255.67 b
(204.48)

52.72 bc
(377.54)
41.82 c
(278.80)
67.49 b
(511.32)

73.13 b
(14.37)
72.85 b
(13.93)
73.83 b
(15.47)

0.73 b
(23.73)
0.75 b
(27.12)
0.76 b
(28.81)

84.76 b
(41.15)
86.97 b
(44.83)
87.62 b
(45.91)

Pj
Bs
PB

*Values in the same columns followed by the same letters are not significantly different (P>0.05) according to the
Duncans Multiple Range Test (DMRT).
** Values in parenthesis ( ) indicating the percentage of increases when compared to control (%).

in the three inoculation treatments. Content of nitrogen

in the leaf of rice treated with PB was 7.6%, which was
significantly (P<0.05) higher than that of control. The
contents of phosphorus and potassium in the leaf of rice
treated with P. agglomerans were also significantly
(P<0.05) higher than that of control. Treatments with P.
agglomerans increased the content of nitrogen,
phosphorus and potassium by 59.09% to 72.72%,
57.14% to 78.57%, and 33.33% to 62.96% respectively
when compared to control. The differences in nutrients
content in the leaf of rice imply the differences in the rate
of nutrient uptakes, which probably resulted from the
effect of P. agglomerans.
Results of present study are similar to those of Biswas
et al. (2000) in which rice plants inoculated with
Bradyrhizobium
sp.
IRBG271
isolated
from
Aeschyneomene fluminensis could increase the uptakes
of nitrogen, phosphorus, and potassium by 27,87%,
19.82%, and 10.97% respectively. Similarly, Shaharoona
et al. (2007) reported that treatment of wheat seed with
the suspension of Burkholderia caryophylli ACC7 at
9
density 10 cfu/ml could increase the uptakes of nitrogen,
phosphorus, and potassium by 39.39%, 32.82%, and
28.38%.
In the present study, the chlorophyll contents of rice
treated with Pj, Bs and PB were 38.25, 39.12 and 39.32
SPAD units, which increased by 23.30%, 26.11%, and
26.76%, respectively when compared to control. The
similar result was obtained by Han and Lee (2005), in
which
the
lettuce
inoculated
with
Rhizobium
leguminosarum cv.viciae 128C56G contained 13.91%
more chlorophyll than un-treated control. Nadeem et al.
(2006) showed that the treatment of corn seed through
soaking with the suspension of rhizobacteria strain S20 at

population density of 10 cfu/ml could increase the

chlorophyll content by 102.22% when compared to untreated control.
The increase of nutrients uptakes by the plants treated
with P. agglomerans compost formulation resulted in the
increase of the growth of leaf, stem and root. The
increase in chlorophyll content may lead to the increase
of photosynthesis rate, and in turn will increase the
accumulation of carbohydrate which is represented by
the increase of NAR and RGR. The increase of
chlorophyll content in the leaf of plants treated with P.
agglomerans may be caused by the ACC-deaminase
activity by P. agglomerans (Teng et al., 2010) which slow
down the degradation of chlorophyll (Silva et al., 2004).
Treatment with P. agglomerans significantly (P<0.05)
increased the fresh and dry weights of shoot, fresh and
dry weights of root, length of stem, diameter of stem, and
leaf area when compared to control (Table 3), however
there is no significant differences (P>0.05) of those
parameters were detected in the three inoculation
treatments. The increase of fresh weights of shoots on
treatments Pj, Bs and PB were 45.79%, 90.25%, and
110.1% respectively when compared to control. The
similar trend was found on the data of dry weight of
shoot, in which treatment PB resulted in the highest
weights among other treatments. This result is similar to
the finding of Sarma et al. (2009), that the treatment with
0.5 g Pseudomonas fluorescens strains R62 dan R81 in
form of powder formulation increased the dry weight of
shoot of Vigna mungo by 110% and 102%, respectively.
Kannan and Ponmurugan (2010) reported that treatment
of rice cultivar CO 43 with Azospirillum brasilense could
increase the dry weight of shoot by 48.73%. Idris et al.
(2009) reported a spectacular result that the treatment

KHALIMI et al.

Figure 2. Root appearances of rice treated with and without P.

agglomerans. C: control, Pj treatment with strain PaJ, Bs:
Treatment with strain BS2a and PB: treatment with the mixture
of strains PaJ and BS2a.

through soil dressing with 30 ml suspension of

8
Stenotrophomonas maltophilia strain KBS9-B (10 cfu/ml)
could increase the dry weight of Sorghum bicolor L.
Moench by 260%.
Treatment with P. agglomerans could also increase the
fresh and dry weights of root. The fresh weight of shoot of
rice treated with Pj, Bs and PB were 175.54 g, 229.06 g
and 252.97 g/hill respectively, which were significantly
(P<0.05) higher than that of control (Table 3). The
increase of fresh weights of shoots on treatments Pj, Bs
and PB were 139.70%, 181.33%, and 204.48%,
respectively. Hasanabadi et al. (2010) reported that the
seed of barley which were soaked in the suspension of
Azospirillum lipoferum and Pseudomonas flourescens
could increase the water content of root by 32.53%. On
other study, Amellal et al. (1998) reported that Pantoea
agglomerans strain NAS206 produced exopolysaccharide
(EPS) which capable of increasing the soil water content
through the increase of soil aggregation. The dry weights
of roots on rice treated with Pj, Bs and PB were also
significantly (P<0.05) higher than that of control. The
increase of dry weight of root on treatments Pj, Bs and
PB were 377%, 278% and 511% respectively. This result
is similar to the report by Khalimi and Suprapta (2011)
that soybean treated with Pseudomonas aeruginosa in
compost formula could increase the dry weight of root by
138.91%. While Kannan and Ponmurugan (2010)
reported that treatment with Azospirillum brasilense on
rice cultivar CO43 increased the dry weight of root by
41.95%.
The increase of fresh and dry weight of roots of rice
treated with P. agglomerans may be resulted from the
increase of the number of lateral root when compared to
control (Figure 2). Pantoea agglomerans was reported to

245

produce indole-3-acetic acid (IAA) which can suppress

the growth of primary root, while stimulate the lateral root
and increase the number of root hairs (Bucio et al.,
2007).
The improvement of root system may be
significantly contributed to the water and nutrients
uptakes from the soil that in turn promoted the growth of
plant.
Treatment with P. agglomerans significantly increased
the length and diameter of stem, the leaf area when
compared to control. Treatments Pj, Bs and PB increased
the length of stems by 15.47%, 12.82% and 13.93%
respectively. Study done by Kandasamy et al. (2009)
showed that treatment with Pseudomonas flourescens
strain KH-1 on rice seed increased the length of stem by
48.64%. Treatment by soaking the seed of varnish tree
(Aleurites moluccana) with the suspension of Pantoea
9
agglomerans strain 5/8 at the population density 10
cfu/ml increased the length of branch by 13.15% (Erturk
et al., 2011).
At present study, the diameter of stem of rice on
treatments Pj, Bs and PB increased by 23.72%, 27.11%
and 28.81% respectively. Similar result was shown by
Naserirad et al. (2011) in which the treatment of corn
seed with the suspension of Azotobacter increased the
stem diameter by 26.34%. In addition, the leaf areas of
rice plants were also obviously increased on the
treatment Pj, Bs and PB by 41.14%, 44.82% and 49.91
respectively. Alam et al. (2001) showed that the roots of
rice plants that were soaked for 24 h in the suspension of
Azotobacter
nigricans,
A.
armeniacus,
Bacillus
sphaericus,
B.
megaterium,
Enterobacter,
and
Xanthobacter for 24 h increased the diameter of stem by
11.85%.
One of the mechanisms of P. agglomerans in
promoting the plant growth is through production of
growth hormones and increases the nutrient uptake.
According to Teng et al. (2010), the plant growth can be
promoted by P. agglomerans through the increase of
nitrogen fixation, production of phytohormones,
phosphate solubization and production of enzyme ACC
deaminase. Feng et al. (2006) reported that P.
agglomerans strain YS19 could fix the nitrogen by 1,697
ng/ml of bacterial suspension per hour. Pantoea
agglomerans could produce phytohormones such as
indole acetic acid (IAA), indole -3-aldehyde, indole-3ethanol, abscisic acid (ABA), gibberellic acid, cytokinin
(isopentyladenosine, zeatin riboside, and dihydrozeatin
riboside) which are necessary for plant growth (Cimmino
et al., 2006; Feng et al., 2006). Meunchang et al. (2006)
reported that P. agglomerans could solubilize phosphate.
The success of P. agglomerans to colonize the plant
root is the first step to promote the plant growth. The
rhizosphere is a dynamic environment which is rich in
energy sources from organic compounds secreted by

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Agric. Sci. Res. J

Table 4. Yield components of rice cv. Cicih Medang Selem treated with P. agglomerans
Treatment

C
Pj
Bs
PB

Number of
tillers/hill

Number of
panicles/ hill

Number of
grains/ panicle

10.40 a *
20.10 b
(93.27)**
20.23 b
(94.52)
22.90 c
(120.19)

8.77 a
16.85 b
(92.13)
17.55 b
(100.11)
18.50 b
(110.95)

163.62 a
183.18 bc
(11.95)
180.52 bc
(10.32)
204.44 c
(24.95)

Dry weight of
1,000 grains
(g)
26.73 a
27.03 a
(1.12)
27.11 a
(1.42)
27.21 a
(1.79)

Percentage of
pithy grain/
panicle
86.46 a
93.84 b
(8.54)
94.43 b
(9.22)
94.92 b
(9.78)

Weight of
Grains/hill (g)*
38.71 a
82.85 b
(114.03)
90.71 b
(134.33)
98.39 b
(154.17)

*Values in the same columns followed by the same letters are not significantly different (P>0.05) according to the Duncans
Multiple Range Test (DMRT). Water content was 13% measured with Grain Moisture Tester Model Riceter m5, Serial number
AE36317. Kett Electric Laboratory, Tokyo).
** Values in parenthesis ( ) indicating the percentage of increases when compared to control (%).

plant root. This habitat is favorable to the P. agglomerans

and other rhizobacteria. The root exudates of rice
containing amino acids such as tryptopan, glucuronic
acid, and carbohydrates such as arabinose, mannose,
galactose, glucose, and xylose (Naher et al., 2008).
Amino acids and organic compounds secreted by rice
plants are converted into IAA by P. agglomerans. The
IAA released by P. agglomerans is utilized by the plant
for root elongation and stimulate the formation of lateral
root through ACC deaminase cycle. In the ACC
deaminase cycle, IAA reacts with S-adenosylmethionine
(SAM) to form 1-aminocyclopropane-1-carboxylate (ACC)
and the formation of SAM is facilitated by ACC syntase
enzyme (Husen et al., 2008). Furthermore, the ACC in
the plant is used by plant and P. agglomerans. The ACC
is a precursor in the production of ethylene. Process of
ethylene production resulted from ACC oxidation which is
facilitated by ACC oxidase (Van Loon and Bakker, 2003).
Ethylene is used by the plant for root elongation. On the
other hand, the ACC is also used by P. agglomerans to
support its metabolism. This bacterium converted ACC
into ammonia and -ketobutyrate and this process is
catalyzed by ACC deaminase produced by P.
agglomerans. Utilization of ACC by P. agglomerans is
beneficial to the plant, because if the concentration of
ACC is too high, resulted in the high concentration of
ethylene. It has been known that the high concentration
of ethylene may be toxic to the plant. Thus, the P.
agglomerans through the production of ACC deaminase
will limit the production of ethylene to avoid the toxicity of
the plant tissue (Kausar and Shahzad, 2006; Viveros et
al., 2010).
Pantoea agglomerans promotes the plant growth
indirectly through the production of antibiotic pyrrolnitrin,
chitinolytic enzyme and siderophore that can suppress

the growth of plant pathogens (Chernin et al. 1995, Teng

et al, 2010). Ortmann et.al. (2006) reported that
exopolysaccharides (EPS) of P. agglomerans consisting
of galactose, glucose, succinate, and pyruvate reduce the
concentration of H2O2 in rice and corn, thus can be used
as induced systemic resistance agent.
Treatment with P. agglomerans significantly (P<0.05)
increased the number of maximum tillers, number of
panicles per plant, number of grain per panicle, number
of pithy grain, and yield per plant (Table 4). Number of
maximum tiller on control (C) was only 10.40 per hill,
while on rice with treatments of Pj, Bs and PB, the
number of maximum tillers per hill were 20.10; 20.23; and
22.90 respectively. These data indicated that the number
of maximum tillers per hill increased by 93.27% to
120.19% resulted from the treatments of P. agglomerans.
Treatment with formula PB showed the highest number of
maximum tillers per hill, however, the differences with Pj
and Bs were not significant (P>0.05). Similar study was
done by Hussain et al. (2009) and showed that treatment
of rice root with Mesorhizobium ciceri strain CRI-32
increased the number of maximum tiller by 25.34%.
Other study by Meunchang et al. (2006) showed that
treatment with P. agglomerans strain APC157 on rice
plant increased the number of tiller per hill by 20% when
compared to control.
Treatment of rice with P. agglomerans significantly
(P<0.05) increased the number panicles per hill. Number
of panicles per hill on control was only 8.77, while on rice
treated with Pj, Bs and PB, number of panicles per hill
were 16.85, 17.55, and 18.50 respectively (Table 4).
There were the increases of the number of panicles by
92.13% to 110.95% resulted from the treatment of P.
agglomerans. Hussain et al. (2009) reported that
treatment of rice plant with Rhizobium leguminosarum

KHALIMI et al.

strain LSI-30 increased the number of panicles per hill by

27.33%. Hien (2007) showed that treatment of rice seed
with Pseudomonas fluorescens, Bacillus subtilis, and
Bacillus amyloliquefaciens in peat formula on seedbed
increased the number of panicles per hill by 22.03%.
Treatments with Pj, Bs and PB increased the number of
grain per panicle by 11.95%, 10.32% and 24.95%
respectively (Table 4). Khorshidi et al. (2011) reported
that treatment of rice seedlings with suspension of
Azospirillum lipoferum for 12 h increased the number of
grain per panicle by 5.28%. Hussain et al. (2009) used
Rhizobium phaseoli to soak the rice seedling, and
showed that this treatment increased number of grain per
panicle by 29.21%. However, treatment with P.
agglomerans did not significantly increase the weight of
1,000 grains. This result similar to that of Alam et al.
(2001) in which treatment of rice root with the
suspensions of
A. nigricans, A. armeniacus, B.
sphaericus,
B.
megaterium,
Enterobacter,
and
Xanthobacter for 24 h did not significantly affect the
weight of 1,000 grains. Khorshidi et al. (2011) also
reported that treatment of rice seedling with the
suspension of Azospirillum lipoferum for 12 h did not
significantly affect the weight of 1,000 grains. This
situation might be explained by the fact that the weight of
1,000 grains is a stable character of the rice cultivars.
Percentage of pithy grain per panicle produced by rice
treated with P. agglomerans significantly (P<0.05) higher
than that of untreated rice plants (control). Rice plants
treated with PB produced the highest percentage of pithy
grain among other treatments (Pj and Bs), however, the
differences among them were not significant (P>0.05).
Treatment with P. agglomerans significantly (P<0.05)
increased the yield per hill, in which the weight of grains
per hill on rice treated with Pj, Bs and PB were 82.85 g,
90.71 g and 98.39 g respectively, which were 114.03%,
134.33% and 154.17% higher than that of control (Table
4). There were no significant differences on the yield per
hill among rice treated with Pj, Bs and PB. Khorshidi et
al. (2011) reported that treatment of rice seedling with the
suspension of Pseudomonas fluorescens for 12 h
increased the yield by 33.3%. Naserirad et al. (2011)
showed that treatment of corn seed with the suspension
of Azotobacter and Azospirillum increased the yield by
23.66%.
Although the treatments with P. agglomerans as single
isolate (Paj or BS2a) and as a mixture between the two
isolates (PaJ and BS2a) resulted in non-significant
differences, however in general treatment with P.
agglomerans as a mixture between PaJ and BS2a
resulted in better rice growth and higher yield than that of
single isolate of PaJ or BS2a. This result suggested a
possible synergistic relation between the two strains. The
detail mechanisms of this synergy need to be studied in
future.

247

CONCLUSION

Application of P. agglomerans strains PaJ and BS2a as a

mixture or as single strain effectively promoted the rice
growth and increased the yield. Rice plants treated with
P. agglomerans as a mixture of two strains, PaJ and
BS2a exhibited better growth and produced higher yield
than those of rice plants treated with P. agglomerans as
single strain, PaJ or BS2a. This result suggested that a
synergy may happen between P. agglomerans strain PaJ
and BS2a.

ACKNOWLEDGEMENT
We would like to express our gratitude and appreciation
to the Udayana University for providing the research
grant Number 0791/023-04.2.01/20/2011 in the fiscal
year 2011 to support this study.

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