INDIAN JOURNAL OF APPLIED MICROBIOLOGY

28 R AMANATHAN
Copyright  2012 Indian Association of Applied Microbiologists, Chennai, India
Volume 15 Number 1 January-June 2012, pp. 28-34.

Response of Sesame (Sesamum indicum L.) to Single

and Dual Inoculation with Azospirillum brasilense
and Glomus fasciculatum at Different NPK Levels

N.Ramanathan*
Department of Microbiology, Faculty of Agriculture, Annamalai University, Annamalai nagar,
Tamilnadu- 608 002, India.

Abstract: Response of sesame (Sesamum indicum L), an important oil seed crop, to the inoculation of
Azospirillum brasilense and arbuscular mycorrhizal (AM) fungus Glomus fasciculatum was studied
in a field experiment. The single and dual inoculation effects of A. brasilense and G. fasciculatum on
growth and yield of sesame cultivar TMV 6 was studied at different NPK (Nitrogen : Phosphorus:
Potash) levels viz,. recommended dose (35: 23: 23 NPK Kg ha -1), 75 % of recommended dose, 50 % of
recommended dose. The dual inoculation of both the fungi caused significant increase in plant height,
shoot and root dry matter production and seed yield over un-inoculated control and as well as single
inoculation of A. brasilense or G .fasciculatum at each NPK level. The sesame seed yields at
75 % (589.5 Kg ha -1) and 50 % (512.0 Kg ha -1) NPK levels with dual inoculation were greater than the
yields obtained at un-inoculated 100% (580.1 Kg ha -1) and 75 % (497.7 Kg ha -1) NPK respectively.

Key words: Sesamum indicum, Azospirillum brasilense, Glomus fasciculatum Arbuscular mycorrhiza,
NPK

Introduction
Sesame, (Sesamum indicum L.) is the most ancient oil seed crop of tropics and warm temperate
regions. India is the largest producer of this oil seed crop of dry farming system. With the present

INDIAN JOURNAL OF APPLIED MICROBIOLOGY Vol. 15 No. 1 Jan.-Jun. 2012

*Author for Correspondence.E-mail: nrmicro@gmail.com
 Response of Sesame to Azospirillum brasilense and Glomus fasciculatum 29

growth rate in oil seed production in India, the demand-supply gap is unlikely to be narrowed down.
Hence, the production and productivity of oil seeds in India should be accelerated further to meet the
future demands by adopting new technologies. Sesame is mostly grown in sub-marginal lands and
characterized by low yield. Biological nitrogen fixation (BNF) has become important in the farming
systems because this process diminishes the need for expensive chemical fertilizers which are
contraindicated with numerous health and environmental problems. The extensive exploitation of
BNF would provide economic benefits to small farmers, avoiding all malign influences of chemical
fertilizers. Microbial inoculants offer an opportunity to introduce eco-friendly, renewable nutrient
sources in crop production.
Azospirillum, a diazotroph (atmospheric nitrogen fixing bacterium) is widely distributed in the
rhizosphere of several tropical crops. Of the various rhizosphere – associated bacteria, Azospirillum
species are probably the most studied and appear to have significant potential for commercial
application [1]. The association between Azospirillum and roots of plants augment the growth and
yield of crops of agronomic importance has been reported by many workers [2-6]. The inoculation of
Azospirillum benefit the crop plants not only through nitrogen fixation but also by the production of
indole acetic acid [7,8]. It is assumed that the bacteria affect plant growth mainly by the production of
plant growth promoting substances, which leads to an improvement in root development and an
increase in the rate of water and mineral uptake [9]. The following plant responses to Azospirillum
inoculation in agriculture crops have been well documented: increase the germination percentage and
height of the seedlings, increase the total number and length of adventitious roots and root hairs,
increase the hydraulic conductivity of the root system, increase the height of the shoot and dry matter
production and yield of the crop plants [10]. The percentage of success due to Azospirillum inoculation
in promoting the yield of agriculturally important crops is of 60-70 % with statistically significant
increase in yield in the order of 5 – 30% [11].
The arbuscular mycorrhiza (AM) is a ubiquitous symbiosis in the world of ecosystems, probably
occurring in two-thirds of vascular plant species. The mycorrhizal hyphae take up nutrients such as
phosphorus, translocate them to host plant, thereby directly increase host nutrient acquisition.
The beneficial influences of AM fungi on plant growth and yield, tolerance to drought and salinity,
resistance to pathogens have been extensively documented [12]. The external hyphae are able to
absorb phosphorus directly from available phosphorus pools in the soil and translocate it to the host
root [13].
Among the microorganisms colonizing the rhizosphere, AM fungi occupy a unique ecological
position as they exist partly inside and partly out side the root and mobilize mineral nutrients especially
phosphorus. Evidently, both Azospirillum and AM fungi are present in the same cortical region of the
root, making the direct interaction possible among three symbionts: Azospirillum – AM fungus –
Host plant. The synergistic interaction of Azospirillum and AM fungi on growth and yield of various
crop plants have been reported [14-17]. This tripartite system contributes significantly to the economy
of nitrogen and phosphorus in natural environments particularly in infertile soil.

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30 R AMANATHAN

Materials and Methods

A field experiment was conducted during rabi season (Feb – May) at Annamalai University Experimental
farm to study the response of sesame cultivar (cv) TMV 6 to single and dual inoculations of Azospirillum
brasilense and Glomus fasciculatum (AM fungus) at different levels of NPK (100 %, 75 %, 50 % of
recommended dose). The microplots of size 6 m x 4 m were laid in split plot design. The main plot
treatments of NPK fertilizers viz., recommended dose (35: 23: 23 NPK Kg ha -1) , 75 % of recommended
dose (26.25:17.25: 17.25 NPK Kg ha -1), 50 % of recommended dose (17.50: 11.50: 11.50 NPK Kg ha -1)
were applied basally. Nitrogen was applied as urea, phosphorus as single super phosphate and
potassium as muriate of potash.
The sub-plot treatments were,
i. control (uninoculated)
ii. A. brasilense inoculation
iii. G. fasciculatum inoculation
iv. Dual inoculation A. brasilense and G. fasciculatum
The surface sterilized seeds of sesame cv TMV 6 were inoculated with A. brasilense (strain AS5,
isolated from sesame roots in semi solid nitrogen free bromothymol blue malate medium). A. brasilense
AS 5 culture was grown in nutrient broth for 72 h at 30 °C. The culture broth containing 1010 cells ml1
was used at the rate of 100 ml per kg of sesame seed. The shade dried seeds were mixed with sand and
sown. The AM culture of G. fasciculatum was multiplied in sorghum (Sorghum vulgare Pers) raised in
garden soil. The top soil along with root bits constituted the mycorrhizal inoculum for field application.
The mycorrhizal inoculum containing infective propagules such as chlamydospores (700 per 100 g),
root bits with inter and intracellular hyphae, vesicles and arbuscules of G. fasciculatum was placed 5
cm below soil surface as a thin uniform layer before sowing the seeds. The soil based AM inoculum
was used @ 5 kg per microplot. The seeds of sesame (inoculated and uninoculted as per the treatment)
were sown in rows 30 cm apart from each other. A total of 180 plants per microplot were maintained
under irrigated condition. Crop production technologies were followed as per the schedule [18].
The growth parameters such as plant height, shoot dry matter production and root dry weight
were recorded on 30th, 60th and 90th days after sowing (DAS). The yield components such as, number
of capsules per plant, 1000 seed weight and seed yield were recorded at 90 DAS (at harvest).

Results and Discussion

The effects of single and dual inoculation of A. brasilense and G. fasciculatum at different NPK levels on
growth parameters - plant height, shoot dry matter production and root dry weight of sesame cv TMV 6
at 90 DAS are presented in the Table 1. The effect of single microbial inoculations on these growth
parameters was found to be superior over uninoculated control. There was an increase in plant height
and dry weight of shoot and root due to single inoculation of either A. brasilense or G. fasciculatum or

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 Response of Sesame to Azospirillum brasilense and Glomus fasciculatum 31

Table 1. Effect of A.brasilense and G. fasciculatum inoculation on plant height,

shoot dry matter and root dry weight at different NPK levels

Plant height in cm Shoot dry matter g plant -1 Root dry weight g plant -1
Treatment
NPK level NPK level NPK level
Inoculations 100% 75% 50% Mean 100% 75% 50% Mean 100% 75% 50% Mean
Control
(uninoculated) 107.6 99.4 88.7 98.6 14.31 12.97 10.68 12.65 3.63 3.27 2.87 3.26
Azospirillum
brasilense 118.1 112.6 102.3 111.0 16.17 14.94 12.72 14.61 4.50 4.03 3.30 3.94
Glomus
fasciculatum 112.6 106.0 95.1 104.2 15.32 13.78 11.83 13.64 4.03 3.63 3.07 3.58
Azospirillum
brasilense +
Glomus
fasciculatum 124.8 118.9 110.5 118.1 16.94 15.76 13.97 15.56 4.83 4.53 3.83 4.40
Mean 115.8 109.0 99.1 15.69 14.36 12.30 4.25 3.87 3.27
MT x ST SED 1.20 0.31 0.28
CD (p=05) 2.42 0.62 0.56

Table 2. Effect of A. brasilense and G. fasciculatum inoculation on number of capsules,

seed weight and seed yield at different NPK levels

Capsule number plant -1 1000 seed weight Seed yield kg ha -1

Treatment
NPK level NPK level NPK level
Inoculations 100% 75% 50% Mean 100% 75% 50% Mean 100% 75% 50% Mean
Control
(uninoculated) 81.2 74.4 65.7 73.8 2.93 2.61 2.57 2.70 580.1 497.7 424.1 500.6
Azospirillum
brasilense 89.8 80.6 74.5 81.6 3.19 2.82 2.70 2.90 626.1 551.2 463.5 546.9
Glomus
fasciculatum 85.2 77.8 69.3 77.4 3.13 2.67 2.62 2.81 605.2 529.7 445.2 526.7
Azospirillum
brasilense +
Glomus
fasciculatum 96.7 85.4 78.7 86.9 3.22 3.00 2.90 3.04 648.5 589.5 512.0 583.3
Mean 88.2 79.5 72.2 3.12 2.77 2.70 615.0 542.0 461.2
MT x ST SED 0.48 0.025 2.31
CD (p=05) 0.97 0.050 4.64

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32 R AMANATHAN

their combination (dual inoculation) compared to the uninoculated plants at each of the three NPK levels.
Individually, the performance of A. brasilense was better than G. fasciculatum with respect to the ability
to stimulate plant growth at different NPK levels. Okon et al. [11] had reported the proportionate increase
of beneficial effects of Azospirillum to crops with intermediate levels of fertilizers (NPK).
In the present study, there was 16, 20 and 25 per cent increase in plant height, 18, 22 and 31 per
cent increase in shoot dry matter production and 33, 39 and 34 per cent increase in root dry weight of
dual inoculated plants respectively at 100 %, 75 % and 50 % NPK levels compared to their respective
uninoculated control at 90 DAS. Earlier studies had recorded that the dual inoculation of maize with A.
brasilense and G. fasciculatum produced significant growth and dry matter production at different
levels of fertilizers. Similar findings of higher dry matter yield due to dual inoculation of Azospirillum
and AM fungi had been reported in pearl millet [19], sorghum [20], coffee [21] and wheat [16].
Interestingly in the present study, compared to uninoculated plants at 100 % NPK level, the
plants with dual inoculation at 75 % NPK level recorded significant increase in plant height (growth)
and shoot and root dry weight by 10.5, 10.0 and 25.0 per cent respectively. The beneficial effects of
dual inoculation in augmenting the growth and dry matter production of sesame at 75 % NPK level
compared to uninoculated 100 % NPK has been clearly established in the results of the present study.
The data presented in the Table 2 show that the response of sesame cv TMV 6 to single microbial
inoculations in yield components such as number of capsules per plant, 1000 seed weight, seed yield
was superior over uninoculated control at different NPK levels. In the present study, at 100 % and 75
% NPK levels the increase in capsules per plant, 1000 seed weight and seed yield was up to 19, 10 and
12 and 15, 15 and 19 per cent respectively due to dual inoculation over uninoculated control. Prabakaran
[20] reported that the dual inoculation of sorghum with A. brasilense and G. fasciculatum resulted in
40 per cent increase in grain yield over uninoculated control. Panwar [16] investigated the response of
A. brasilense and G. fasciculatum inoculation to wheat and reported higher grain yield in combined
inoculation with A. brasilense and G. fasciculatum compared with single inoculations.
The sesame seed yield at 75 % (589.5 kg ha1) and 50 % NPK (512.0 kg ha1) levels with dual
inoculation was greater than the uninoculated counterpart at 100% (580.1 kg ha1) and 75 % NPK (497.7
kg ha1) respectively. The results of the present study conducted in the field revealed that the synergistic
effects of combined inoculation of A. brasilense and G. fasciculatum resulted in increased plant
growth and seed yield. It is further established that by dual inoculation to this crop sesame, NPK
fertilizer application can be reduced by 25% without reduction in yield. This can be attributed to the
fact that microbial inoculations of A. brasilense and G. fasciculatum might have augmented the
availability of nitrogen (biological nitrogen fixation) and phosphorus (phosphate mobilization) to the
crop plants thus ultimately increasing the yield. Therefore, the present study suggests that the dual
inoculations of A. brasilense and G. fasciculatum could be considered as potential supplements for
chemical fertilizers in integrated nutrient management.

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 Response of Sesame to Azospirillum brasilense and Glomus fasciculatum 33

Acknowledgement
The author is thankful to the authorities of Annamalai University, Annamalai nagar, for providing the
facilities to carry out this study and granting permission to publish the work.

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