European Journal of Soil Biology 45 (2009) 334340

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European Journal of Soil Biology

journal homepage: http://www.elsevier.com/locate/ejsobi

Original article

Reduction in dose of chemical fertilizers and growth enhancement of sesame

(Sesamum indicum L.) with application of rhizospheric competent
Pseudomonas aeruginosa LES4
Sandeep Kumar a, Piyush Pandey b, D.K. Maheshwari a, *
a
b

Department of Botany and Microbiology, Gurukul Kangri University, Haridwar (U.A.) 249404, India
S.B.S.P.G. Institute of Biomedical Sciences and Research, Balawala, Dehradun (U.A.) 248161, India

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 9 September 2008
Received in revised form
4 April 2009
Accepted 7 April 2009
Available online 19 April 2009
Handling editor: Kristina Lindstrom

Pseudomonas aeruginosa LES4, an isolate of tomato rhizosphere was found to be positive for several plant
growth-promoting attributes like production of indole acetic acid, HCN and siderophore, solubilization of
inorganic phosphate along with urease, chitinase and b-1-3-glucanase activity. In addition, it showed
strong antagonistic effect against Macrophomina phaseolina and Fusarium oxysporum. P. aeruginosa LES4
caused halo cell formation and other morphological deformities in mycelia of M. phaseolina and
F. oxysporum. Root colonization was studied with Tn5 induced streptomycin resistant transconjugants of
spontaneous tetracycline-resistant LES4 (designated LES4tetrastrep) after different durations. The strain
was signicantly rhizospheric competent, as 17.4% increase in its population was recorded in sesame
rhizosphere. Seed bacterization with LES4 resulted in signicant increase in vegetative growth parameters and yield of sesame over non-bacterized seeds. However, application of LES4 with half dose of
fertilizers resulted in growth equivalent to full dose treatment, without compromising with the growth
and yield of sesame. Moreover, the oil yield increased by 33.3%, while protein yield increased by 47.5%
with treatment of half dose of fertilizer along with LES 4 bacterized seeds, as compared to full dose of
fertilizers.
2009 Elsevier Masson SAS. All rights reserved.

Keywords:
PGPR
Chemical fertilizer
Macrophomina phaseolina
Fusarium oxysporum oilseed crop
Sesame

1. Introduction
Sesame (Sesamum indicum L.) is an important oilseed crop, next
to soybean and groundnut but has low yield potential [35]. India
rank rst in the world in area (about 2.47 m ha annually, 40% of the
world) and production (0.74 m tones, 27% of the world). In intensive
cropping system, supplementing soil nutrients by use of chemical
fertilizer is considered inevitable for obtaining optimum yield of
crops, however, their utilization efciency remains low, due to loss
by volatization, denitrication, leaching and conversion into
unavailable forms. Continuous use of chemical fertilizers subverts
the soil ecology, disrupt environment, degrade soil fertility and
consequently shows harmful effects on human health [1] and
contaminates ground water [15]. Presence of chemicals in sesame
had been major impediment in the promotion of sesame export.
Export consignments of sesame are sometimes unsuitable in the

* Corresponding author. Tel.: 91 1334 246 767 (O), 265 469 (R), 91 983 730
8897 (M); fax: 91 1334 246 767 (O).
E-mail addresses: [email protected] (P. Pandey), maheshwaridk@
gmail.com (D.K. Maheshwari).
1164-5563/$ see front matter 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejsobi.2009.04.002

international market due to the presence of pesticide residue,

resulting in loss of revenues [6].
Role of plant growth-promoting rhizobacteria (PGPR) in plant
growth promotion and biological control of soil borne pathogens
has been intensively investigated [17,19]. Integration of PGPR with
traditional inorganic fertilizers in the eld may prove to be effective
means to increase the solubility of insoluble phosphorous ions and
other minerals to plants with simultaneous reduction in diseases
incidence. They take systemic and simultaneous account of environmental aspects, quality of the produce and protability of
agriculture [22]. Alternatively, substitution of chemicals with
bacterial fertilizers and biopesticides, especially blending of
chemical fertilizers with chemical adaptive PGPR is a promising
approach to obtain sustainable fertility of the soil and plant growth
[39]. There are several PGPR currently commercialized whose
growth-promoting activity in crop plants have been demonstrated
in several ways including production of iron-sequestering siderophores and antimicrobial compounds that hinder colonization of
hosts by phytopathogens [40], induction of host systemic disease
resistance, solubilization of precipitated mineral nutrients and for
production of plant growth hormones, thereby enhancing the
plants ability to take up nutrients from soil and increasing yield [8].

S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

Hence, the present work was aimed to blend chemical fertilizers

with effective PGPR to obtain the optimum benets. This strategy
was designed to allow reduction in the dose of fertilizers, along
with an approach to increase productivity of sesame without
associated ecological harm.
2. Materials and methods
2.1. Microorganisms
A number of bacterial strains were isolated using standard
microbiological technique from the rhizosphere of tomato
(Lycopersicon esculantum L.) grown in nutrient decient soil in
wasteland. Healthy and young tomato seedlings were gently
uprooted at Dehradun (Alt. 640 m, Lat. 30 300 4000 N, Long.
77 5201200 E, Rainfall 216 cm) in India. The roots were cut into 2 cm
long segments and vortexed in 25 ml sterilized distilled water for
few minutes. Suitable dilution was plated by serial dilution plate
technique on nutrient agar medium (NAM). The bacterial colonies
were isolated and maintained on NAM slants at 4  C. The isolates
were characterized for direct and indirect plant growth-promoting
(PGP) activities including solubilization of inorganic phosphate, IAA
production, and HCN production along with antagonism against
two fungal pathogens. Seven isolates were selected and identied
on the basis of morphological, physiological and a biochemical
characteristic according to Bergeys manual of determinative
bacteriology [14], and compared against Pseudomonas aeruginosa
MTCC-1934, Pseudomonas putida MTCC-102 and Pseudomonas
uorescens MTCC-103 as standard strains. All isolates were maintained on tryptic soy agar medium (TSM) at 4  C for further use.
Macrophomina phaseolina and Fusarium oxysporum were
procured from culture collection laboratory, Department of Botany
and Microbiology, Gurukul Kangri University, Haridwar, India. Pure
culture of fungal colonies was maintained on potato dextrose agar
(PDA) slants at 4  C for further use.
2.2. Screening for plant growth-promoting attributes
Phosphate solubilization [38], IAA production [12], HCN
production [23], siderophore production [32], and chitinase and
b-1-3-glucanase activities [7] were determined as per standard
protocols. Antagonistic activity of isolates against phytopathogens
was determined according to Skidmore and Dickinson [36].
2.3. In vitro antagonism
Antagonistic properties of bacterial strains were tested against
two fungal pathogens M. phaseolina and F. oxysporum causing
charcoal rot and wilt diseases on sesame using a dual culture
technique as described previously [19]. Agar blocks (5 days old,
5 mm dia.) containing 5 days old mycelia were placed in four
corners of a Petri plate, and inoculated with loopful culture (24 h
old) of bacterial strain, spotted 2 cm apart from the fungus. These
plates were incubated at 28  C. Plates inoculated with only fungal
agar blocks served as control. Growth inhibition was calculated by
measuring the distance between the edge of bacterial and fungal
colonies, and percent inhibition was calculated by following
formula:

Growthinhibition C  T=C100
where C radial growth of fungus in control, T radial growth of
fungus in dual culture. Fungal hypha surrounding zone of inhibition, and from control plates were observed under the microscope
by standard procedure.

335

2.4. Seed bacterization

The certied seeds of sesame (S. indicum L. cv. ST-1) were
procured from Center for Biotechnology, Jamia Hamdard University, Hamdard Nagar, New Delhi, India. Seed bacterization was done
by the method of Weller and Cook [41]. Seeds were surface sterilized with 95% alcohol for 30 s, followed by 0.1% (w/v) HgCl2 for
12 min and then washed with sterile distilled water for 56 times.
These germinated seeds were dried under sterile air stream. 24 h
old culture of LES4 was centrifuged at 7000 rpm for 15 min at 4  C.
The pellets were retained and re-suspended in sterile distilled
water to obtain a population density of 108 cfu ml1 and the cell
suspension was mixed with 1% carboxymethyl cellulose (CMC)
solution in ratio of 1:0.5. Slurry thus obtained, was coated on the
surface of germinated seeds. The seeds coated with 1% CMC slurry
without LES4 served as control. The bacterized seeds were dipped
in known volume of sterile water and cfu were counted on TSM for
standardizing the inoculum. The population of LES4 was recorded
by dilution plate technique as 108 cfu seedling1.
2.5. Root colonization
Root colonization of P. aeruginosa LES4 was studied by quantitative analysis of population dynamics in the rhizosphere of sesame
using antibiotic resistant marker strain. Tetracycline-resistant
strain of LES4 was isolated on TSM, containing 100 mg l1 of
tetracycline (P. aeruginosa LES4tetra). LES4tetra was engineered for
streptomycin resistance (100 mg l1) (designated LES4tetrastrep)
with Tn5 delivery suicide vector pGS9 [33] in donor Escherichia coli
strain WA803 (pGS9) as described [18]. Sesame plants emerged
with bacterized seeds was sampled after 30, 60, 90 and 120 DAS,
and bacterial population on the roots were measured. Root
adhering soil particles were carefully removed and root was cut
into 1 cm long segments, which was vortexed in known volume of
sterile water to release root-associated bacteria. Suitable dilutions
of the suspension were plated on TSM containing tetracycline and
streptomycin (100 mg l1 each) to enumerate the bacterial
population.
2.6. Field trial
Field trials of sesame were carried in sandy loam soil (80.3%
sand, 6.5% silt, 7.7 clay, total organic C 0.0923%, pH 6.8 having 35%
water holding capacity). The recommended dose of chemical
fertilizer for sesame crop was 120 kg ha1 nitrogen, in three split
doses of urea, and 30 kg ha1 phosphate in the form of Diammonium phosphate (DAP) and 30 kg ha1 potassium in the form
of Murate of potash (MoP) in single doses. The combination of NPK
was N404040P30K30. Seeds bacterized with P. aeruginosa LES4 and
non-bacterized seeds were sown on randomized block design
(RBD) in 7 sets of treatments with three replicates of each treatment, (i) seeds coated with P. aeruginosa LES4, (ii) P. aeruginosa
LES4 half dose of chemical fertilizers, (iii) half dose of chemical
fertilizers, (iv) full dose of chemical fertilizers, (v) seeds coated with
CMC slurry only without any fertilizer and bacteria (control). The
crop was irrigated three times (including one pre-sowing irrigation) at different critical stages, i.e. at owering/capsule formation
and seed lling. Seed germination rate (%) was noted on 15 days
after sowing (DAS). Vegetative growth parameters including
biomass accumulation, root and shoot lengths, leaf area were
recorded at 30, 60, 90 and yield attributes were recorded after 120
DAS on harvesting the crop. The experiment was conducted for two
consecutive years and data are presented as mean. The data were
analyzed statistically by using analysis of variance (ANOVA) to nd
out signicance at 1% and 5% levels [9].

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S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

2.7. Estimation of oil and soluble protein content in sesame seeds

Oil content is dened as the whole of the substances extractable
by n-hexane under specied conditions. The rapid gravimetric
determination of oil content was done by cold percolation as
described by Kartha and Sethi [16]. The oil content was expressed in
percentage and yield in kg ha1. Soluble protein content of seeds
collected from eld trials was estimated according to the method
given by Bradford [4], after precipitation with trichloroacetic acid
using bovine serum albumin (BSA) as standard.
3. Results
3.1. Isolation of microorganisms
Among the seven uorescent pseudomonads, LES4 was selected
for further studies on the basis of plant growth-promoting
attributes. The isolate LES4, identied as P. aeruginosa was Gramnegative, non-spore forming rod, aerobic, motile and formed small,
round colonies with smooth margins on NAM plates after 24 h of
incubation at 28  1  C. The strain was positive for catalase, oxidase,
urease and indole production and was negative for starch hydrolysis and mannose utilization (Table 3).
3.2. Plant growth-promoting attributes
LES4 and LES7 were found to have excellent phosphate solubilization activity. LES4 formed relatively large zone of clearing on
Pikovskayas medium. Most of the pseudomonads produced IAA
except LES2 and LES5. All pseudomonads produced siderophore
while LES3 and LES4 produced large halos around their colonies on
CAS agar medium. HCN production was evidenced by change in
colour of lter paper from yellow to reddish brown after 3 days of
incubation in all the pseudomonads except LES2 and LES6. LES4 and
LES7 were positive for both chitinase and b-1-3-glucanase activities. LES4 was more effective, showed excellent zone of inhibition
than that of LES7 in dual culture assay against M. phaseolina and F.
oxysporum (Table 4).
3.3. In vitro antifungal activities
P. aeruginosa LES4 strongly inhibited the growth of M. phaseolina
and F. oxysporum and growth inhibition was maximum after 5 days
of incubation. Increase in the incubation time corresponded with
the increase the zone of inhibition up to 5 days, thereafter the
growth of mycelia toward the interaction zone ceased, and the
mycelia gradually lost vigour. P. aeruginosa LES4 caused 85%
inhibition of M. phaseolina and 67% of F. oxysporum in their growth
respectively in comparison to control (Table 1). P. aeruginosa LES4
caused halo cell formation, mycelial deformities and hyphal tip

Table 1
Antagonistic effect of P. aeruginosa strain LES4 on dual culture plate at 28  1  C
against M. phaseolina and F. oxysporum.
Fungal
pathogen

Incubation
(h)

Growth in dual
culture (mm)

Growth in
control (mm)

Growth
inhibition (%)

M. phaseolina

48
72
96
120

21.0
22.3
22.7
22.9






0.08
0.04
0.06
0.07

41.2
51.4
60.7
81.9






0.05
0.04
0.08
0.08

49.0
56.6
62.0
72.0

F. oxysporum

48
72
96
120

20.0
21.1
21.6
21.8






0.08
0.09
0.05
0.09

40.0
42.5
51.7
71.8






0.04
0.08
0.11
0.12

50.0
50.3
58.2
69.6

degradation of M. phaseolina and F. oxysporum (Figs. 1 and 2). Also,

sclerotial and conidial development was ceased to be arrested.
3.4. Root colonization
The antibiotic resistant marker strain LES4tetrastrep showed
positive root colonization of sesame. 17.44% increase in the population of LES4tetrastrep in sesame rhizosphere was observed
between 30 and 120 DAS (Table 2). The nal population of P. aeruginosa LES4tetrastrep was highest in treatment receiving half dose
of fertilizers, as compared to treatment of full dose of fertilizer.
Population of standard strain P. aeruginosa MTCC-1934 also
increased tenfold when applied with half dose of fertilizer between
30 and 120 DAS.
3.5. Field trial experiment
Seeds bacterized with P. aeruginosa LES4 showed enhancement
in seed germination and seedling emergence, in all the treatments
as compared to control. 98% seed germination was recorded in
bacterized seeds with low dose of chemical fertilizers, which was
higher as compared to full dose of fertilizers (90.1%). All vegetative
plant growth parameters were also increased progressively after
30, 60 and 90 DAS as compared to control. Most pronounced
positive interactive effect in all the vegetative parameters and yield
components was observed in treatment of bacterized seeds with
half dose of chemical fertilizers (i.e. N202020P1515K15) where
30.6% increase in number of capsules per plant and 22.9% increase
in shoot fresh weight were obtained, as compared to half dose of
fertilizer treatment (Table 5). Data of growth enhancement of
sesame, obtained with application of full dose of fertilizer were
almost similar to treatment receiving bacterized seeds with half
dose of fertilizers (Table 6).
3.6. Oil and soluble protein content in sesame seeds
The percent oil content of seeds was almost similar in all
treatments; however, a small increase of about 10% oil content was
recorded with treatment of half dose of fertilizer plus LES4, as
compared to control (Table 6). However, the oil yield increase by
33.3% with treatment of half dose of fertilizer with LES bacterized
seeds, as well as full dose of fertilizers, compared to treatment of
reduced dose of fertilizer only. The protein yield increased by 47.5%
in treatments where LES4 was applied in combination with half
dose of chemical fertilizers as compared to full recommended dose
of chemical fertilizers.
4. Discussion
P. aeruginosa LES4 produced IAA, siderophore, HCN and solubilized inorganic phosphate and showed strong antagonism against
two dreaded fungal pathogens, M. phaseolina and F. oxysporum. It is
well known that the benecial effects of plant growth-promoting
rhizobacteria are attributed to the production of diverse metabolites including siderophores, hydrocyanic acid (HCN), IAA and other
associated activities such as good phosphate solubilization and
competition in soil and root colonization [8]. In fact, similar to LES4,
there have been several reports of pseudomonads producing IAA
[2,3,1012], HCN [3,1012], siderophore [41,42], and solubilization
of inorganic phosphates [20,26,5]. Presence of wide array of attributes made LES4 a suitable candidate as PGPR. LES4 also released
chitinase and b-1,3-glucanase in the medium, and inhibited growth
of M. phaseolina and F. oxysporum. It is well known that chitin is
main component of cell wall of M. phaseolina. Similarly, cell wall of
the F. oxysporum composed of mostly of 47% chitin with 14%

S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

337

Fig. 1. Post-interaction morphological changes in F. oxysporum due to P. aeruginosa LES4, (A) hyphal tip degradation, (B) halo cell formation, (C) cytoplasm coagulation, (D) hyphal
perforation, (E) digestion of fungal cell wall as compared to (F) control (bar 10 mm).

laminarin. Therefore, their role in lysis of the cell wall and hence
biocontrol cannot be ruled out. This fact was supported by the
morphological changes in hypha, observed after interaction of
M. phaseolina and F. oxysporum with LES4.
An integrated approach wherein microbial inoculants applied
along with reduced level of fertilizers so as to obtain better
growth and yield is essentially required [24,37]. The main
objective of this work was to access the efciency of PGPR to
reduce the use of chemical fertilizers, without compromising with
the growth and yield of sesame. The results of this study suggest
that P. aeruginosa LES4 in combination with half dose of fertilizers
signicantly inuenced the yield viz. number of capsule per plant,
number of seeds per capsule as well as seed weight and oil yield
of S. indicum L. Earlier also, a strong positive correlation between

yield component and seed yield have been reported by several

investigators in other crops [30]. Higher concentrations of
chemical fertilizers have been reported as lethal for bacterial
growth, but found to stimulate its growth at low concentrations
[19]. This may be one of the reasons for growth enhancement of
host plant by PGPR in presence of lower concentration of fertilizer
in soil.
The amount of protein concentration increased in all the
treatments of biofertilizers in combination with reduced dose of
fertilizers in comparison to control. Application of Azospirillum sp.
reduced the nitrogen requirement by 50% in S. indicum [29]. This
was in accordance to earlier reports with crops like soybean where
signicant increase in dry matter, seed protein, oil and yield of
soybean on inoculation with rhizobia and phosphate solubilizing

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S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

Fig. 2. Post-interaction morphological changes in M. phaseolina due to P. aeruginosa LES4, (A) sclerotial cease in M. phaseolina, (B) halo cell formation, (C) mycelial deformities in
M. phaseolina as compare to (D) control (bar 10 mm).

bacteria in combination with chemical fertilizers was reported

[21,43]. Contrary to this, Poul and Savithri [28] reported that the
growth and yield of sesame with application of half dose of urea in
combination with biofertilizers was less as compared to dose of
inorganic fertilizer. However, in present study, P. aeruginosa LES4
resulted equivalent yield as well as growth when applied with half
dose of fertilizers, as compared to application of full dose of
fertilizers, which may be because the difference in PGPR used.
Strong root colonization ability of LES4 lies in it being the
successful efcient colonizer in the rhizosphere of sesame. The use of
genetic markers such as intrinsic levels of resistance to various antibiotics is one of the simple and rapid methods for strain identication
in rhizosphere [25,27], and hence utilized in this study. Combined
resistance against two broad range antibiotics in introduced marker
strain LES4tetrastrep allowed no background population of other
strains, which was further assured by checking the phenotypic and
physiological characteristics. The LES4tetrastrep showed efcient
root colonization after different time intervals of observation (i.e. 30,
60, and 90 days) and enhanced plant growth and growth yield. The
population density increased up to 60 days of inoculation, which

became almost stable thereafter. It has been suggested that bacteria

that attain CFU of about z103 per gram or higher on root mass can be
considered as good colonizers [31], and we obtained population of
LES4tetrastrep of z107. The continued presence of P. aeruginosa
LES4tetrastrep for 90 days in soil showed that it had reached
homeostasis after undergoing exchange with indigenous microora
and is not affected by the active and passive processes restricting soil
community. These results were similar to root colonization of
P. aeruginosa NBRI2650 [25], however they studied the colonization
up to 60 days only. Earlier, P. aeruginosa has been reported as potential
biocontrol agent and simultaneously enhanced plant growth and root
colonization of various vegetables and cereals without showing any
deleterious effects of plants [13,27,34].
On the basis of present ndings, it may be concluded that
P. aeruginosa LES4 contains large number of plant growth-promoting
attributes, and its application contributed in enhancement of
sesame growth leading to better yield. In addition, its ability to
reduce the chemical requirement for obtaining optimum yield of
sesame appears to be of great ecological and economic importance,
with possibilities of an efcient bioinoculant.

Table 2
Root colonization of S. indicum L. by P. aeruginosa LES4 and standard strain MTCC-1934.
Bacterial population (log10 cfu)
Treatments
P.
P.
P.
P.

aeruginosa
aeruginosa
aeruginosa
aeruginosa

30 DAS
LES4
MTCC-1934
LES4 half dose fertilizers
MTCC-1934 half dose of fertilizers

Data are mean  SD of two years.

6.25 
5.27 
6.75 
5.88 

60 DAS
0.12
0.09
0.12
0.12

6.35
5.34
6.86
5.89






90DAS
0.14
0.11
0.11
0.14

7.10 
6.16 
7.33 
6.87 

120 DAS
0.13
0.14
0.17
0.19

7.34
6.72
7.53
6.90






0.12
0.18
0.19
0.16

S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

339

Table 3
Morphological, physiological and biochemical characters of uorescent Pseudomonas strains with standard strains.
Characteristics

LES1

LES2

LES3

LES4

LES5

LES6

LES7

MTCC-102

MTCC-103

MTCC-1934

Gram reaction

Growth at
4 C
41  2  C
Generation time (h)
Fluorescent diffusible pigment
Endospore
Capsule
Non-uorescent non-diffusible pigment
PHB accumulation
Motility
Urease
Oxidase
Catalase
VP test
MR test
H2S production
Gelatin hydrolysis
Starch hydrolysis
Arginine hydrolysis
Citrate utilization

1.2
(bg)









1.3
(y)









1.4
(bg)









1.2
(bg)









1.4
(y)










1.3
(bg)









1.2
(bg)










1.3
(y)










1.3










1.2
(bg)









Utilization of carbon as
Glucose
Maltose
Mannose
Trehalose
Mannitol
Ribose
Meso-Inositol





























Abbreviations: , positive; , negative; y, yellow; bg, blue green; VP, Voges Proskauer, MR, methyl red; H2S, hydrogen sulde; PHB, poly-b-hydroxylbutyrate; P. putida MTCC102; P. uorescens MTCC-103 and P. aeruginosa MTCC-1934.
Table 4
Plant growth-promoting and antifungal properties of isolated uorescent Pseudomonas species.
Strains

IAAA

Phosphate solubilizationB

SiderophoreC

HCND

ChitinaseE

b-1,3-glucanaseF

Antagonism against
M. phaseolina

Antagonism against
F. oxysporum

LES1
LES2
LES3
LES4
LES5
LES6
LES7
MTCC-1934
MTCC-103
MTCC-102

































Abbreviations: A, IAA negative; , IAA positive; B, phosphate solubilization negative; , phosphate solubilization positive; C, absence of halo formation; , small
halos < 0.5 cm wide surrounding colonies; , medium halos > 0.5 cm wide surrounding colonies; , large halos > 1.0 cm wide surrounding colonies; D, HCN negative;
, HCN positive; E, chitinase negative; , chitinase positive small halos < 0.5 cm wide surrounding colonies; , medium halos > 0.5 cm wide surrounding colonies; F, b1,3-glucanase negative, , b-1,3-glucanase positive; G, no inhibition; , 025% inhibition; , 2650% inhibition; , 6085% inhibition against M. phaseolina; P. aeruginosa
MTCC-1934; P. uorescens MTCC-103; P. putida MTCC-102.

Table 5
Interactive effect of Biofertilizers and Integrated Nutrient Management Protocol on the seed germination and growth and yield component of S. indicum L. cv. ST-1 after 120 DAS.
Treatment

T-1
T-2
T-3
T-4
T-5

P. aeruginosa LES4
P. aeruginosa LES4 half
dose fertilizers
Half dose of fertilizers
Full dose of chemical
fertilizers
Control
SEM
CD at 1%
CD at 5%

Germination
(%)

Root
Length
(cm)

Fresh
weight (g)

Dry
weight (g)

Shoot
Length
(cm)

Fresh
weight (g)

Dry
weight (g)

Leaf
area (cm2)

Capsules/
plant

92.0
98.0

16.1
18.7

38.0
45.6

21.1
27.5

165
206

204
247

107.5
128.8

768
875

98.2
128

89.3
90.1

14.0
17.8

38.6
42.8

20.2
26.0

163
200

201
242

97.5
114.5

759
853

98.7
123

76.2
0.732
3.161
2.255

10.2
0.437
1.886
1.346

27.6
0.756
3.264
2.329

14.2
0.247
1.068
0.762

130
0.563
2.432
1.735

146
1.168
5.044
3.599

74.1
0.611
2.640
1.883

569
0.939
4.054
2.893

48.2
1.185
5.119
3.653

Values are mean of 10 randomly selected plants from each set; ns, not signicant; *signicant at 5% LSD; **signicant at 1% level of LSD as compared to control; full dose of
chemical fertilizers N404040, P30, K30; half doses of chemical fertilizers N202020, P15, K15.

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S. Kumar et al. / European Journal of Soil Biology 45 (2009) 334340

Table 6
Interactive effect of Biofertilizers and Integrated Nutrient Management Protocol on the yield and yield component of S. indicum L. after 120 DAS.
Treatment

T-1
T-2
T-3
T-4
T-5

P. aeruginosa LES4
P. aeruginosa LES4 half dose fertilizers
Half dose of fertilizers
Full dose of chemical fertilizers
Control
SEM
CD at 1%
CD at 5%

Average seeds yield (kg/ha)

1000 seeds weight (g)

2004

2005

Mean

2004

2005

Mean

649
987
719
986
462
0.338
1.459
1.041

661
1011
763
998
476
1.514
6.539
4.666

655
999
741
992
469
1.432
6.183
4.412

2.73
3.01
2.59
2.92
2.40
0.845
0.365
0.260

2.84
3.04
2.65
3.03
2.48
0.115
0.500
0.357

2.78
3.02
2.62
2.96
2.44
0.158
0.685
0.489

Oil content
(%)

Oil yield
(kg/ha)

Protein
content (%)

Protein yield
(kg/ha)

49.6
50.4
47.2
49.6
45.8
1.095
4.729
3.374

370
496
372
495
215
1.309
5.654
4.034

19.0
20.2
18.6
20.0
18.3
1.303
5.626
4.015

136
208
141
203
97
1.105
4.773
3.406

Data are mean of two years. Values of each year are mean of 10 randomly selected plants from each set; ns, not signicant; *signicant at 5% LSD; **signicant at 1% level of LSD
as compared to control; full dose of chemical fertilizers N404040, P30, K30; half doses of chemical fertilizers N202020, P15, K15.

Acknowledgments
The authors wish to thank the TMOP & M, CSIR, New Delhi for
nancial assistance.

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