UM 3rd Report - Innotech Bio Fertilizer

FINAL REPORT

CONSULTANCY PROJECT

TITLE:
Effects of Biofertilizers on Growth, Soil Fertility and Nutrients Uptake of

Oil Palm (Elaeis guineensis) Under Glasshouse Conditions

BY:
PROFESSOR DR LING TAU CHUAN
DR ROSAZLIN BINTI ABDULLAH

WITH
INNOTECH FERTILIZER SDN. BHD

SUMMARY
The use of compound fertilizers (CF) in oil palm plantations poses an enormous threat to the
ecosystem through the degradation of soil and water quality. Thus, oil palm biofertilizers are
potentially more environmentally friendly compared to chemical fertilizer. A glasshouse study
was conducted in Rimba Ilmu, University of Malaya to test the efficiency between the
treatments of compound fertilizer with combination of Innotech Biofertilizer focusing on the
soil fertility, nutrient uptake and the growth of oil palms seedlings. Three dominant strains of
bacteria were identified in the liquid culture of Innotech biofertilizer, namely Bacillus
tequilensis strain 10b, Bacillus amyloliquefaciens strain MPA 1034 and Bacillus cereus strain
JCM 2152. These strains of PGPR have been reported to enhance growth and fix nitrogen with
several non-leguminous crops. Bacillus group of bacteria have been listed as one of the N2-
fixing bacteria associated with the non-legumes nitrogen fixation. Two types of treatments
were designed: [T1] Chemical Fertizer only, NPK Mg (12:12:17:2 + TE) and [T2] 70% NPK
Mg (12:12:17:2 + TE) + 30% Innotech biofertilizer. Plant growth parameters which include
plant height, chlorophyll content, girth size, number of fronds, including the dry weight of
shoots and roots and NPK content were observed in this study. The results indicated that there
was a significant difference between the treatments on the chlorophyll content, girth size and
number of fronds of the seedlings. From this study, the combined fertilization with Innotech
biofertilizer showed a profound effect on the proliferation and development of the roots. The
combined use of Innotech Fertilizer with compound fertilizer showed a positive response in
enhancing the accumulation of nutrients uptake (NPK), shoots and roots biomass and plant
growth parameters in seedlings planted in glasshouse conditions.

Keywords: Plant growth promoting rhizobacteria, oil palm seedlings nursery, biofertilizers,
compound fertilizer.

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1.0 INTRODUCTION

Agriculture sector is considered as one of the economy pillars in developing nations (Zahid
2015). However, the dependence on chemical fertilizers and pesticides in this sector is
detrimental to human consumption and which also cause ecological imbalance (Bhardwaj,
Ansari et al. 2014). According to FAO in 2011, about 175.5 million tons of chemical fertilizer
is used in agriculture to achieve an optimum crop yield. The use of chemical fertilizers on
crops has become a regular practice in conventional agriculture and still relevant to this day,
where farmers apply fertilizers to promote the growth of their crops. In larger plantation areas
such as the oil palm plantation, purchase of these chemical fertilizer constitutes about 40-50%
of the total cost (Ng et al., 2002). In addition, chemical fertilizers such as nitrogenous fertilizers
are expensive and small-scale farmers will not be able to purchase fertilizers for their crops,
forcing them to find cheaper alternatives.

Chemical fertilizers can cause imbalance in the agroecosystem and degradation in the soil
quality. The dependence on chemical fertilizer will also cause air and ground water pollution
resulted from eutrophication, which further effects our aquaculture and water quality
(Shumway, 1990; Tang et al., 2003). Moreover, chemical fertilizers are more resistant in the
environment causing the loss in microbial diversity and population. This practice also weakens
the roots of the crops, making them susceptible to pests and diseases (Ju, Wj et al. 2018)

Therefore, there is a need to replace this conventional agricultural practice by implying a safer
alternative to promote the growth of the plants, without affecting the environment and the
agroecosystem. The effort to reduce the dependence on the chemical fertilizers has been made
through the establishment of biological based organic fertilizers (also known as biofertilizer)
as an alternative (Bhardwaj, Ansari et al. 2014). Biofertilizers can be made up of from soil
bacteria that are beneficial to the plants and it is known as integrated nutrients system where
are nutrients required by the plants are provided by the activity of the below-ground
microorganisms.

Beneficial bacteria such as Plant Growth Promoting Rhizobacteria (PGPR), Nitrogen – fixers,
phosphorus and potassium solubilizers, combined with the use of some beneficial fungi are
often utilized in the production of biofertilizers (Mohammadi et al., 2012). For PGPR, the

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genera belong to the group of rhizobacteria can directly and indirectly promoting the growth
of the crops by the production of nutrients, siderophores that are used for protection by limiting
the availability of Iron to the phytopathogens. Some genera such as Azospirillum sp. and
Bacillus sp. are the symbiotic nitrogen fixing bacteria which efficiently fix the nitrogen in the
nodules of the plants, hence reduces the dependence on nitrogenous fertilizers. (Amir et al.,
2005).

Figure 1. The possible mode of action used by plant growth promoting rhizobacteria (PGPR)
towards growth promotion in plants. The flow and location of nitrogen fixation, phosphorus
solubilization, and siderophore production are shown (Vacheron et al., 2013)
According to Khan et al., 2012, the harmful effects from using chemical fertilizers can be
mitigated using biofertilizers, reducing the global dependence on the hazardous chemical
fertilizers. Such bacteria have been isolated from different part of agricultural lands and studied
intensively before their utilization in the biofertilizers. Some strains of bacteria also undergone
strains improvement via genetic engineering to enhance their specific tasks.
Biofertilizers are environmental-friendly and do not cause any detrimental effects towards the
plant and the agroecosystem, and they are readily available at a cheaper price. One of the
problems caused by the excessive use of fertilizers by the farmers are the soil infertility, but

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with biofertilizers, the fertility of soil can be restored, promoting a better growth of the crops
grown (Mohammadi et al., 2012).
In this study, oil palm (Elaeis guineensis) will be used to study the growth effects of the
biofertilizers application under a glasshouse condition. E. guineensis is one of the major
plantations in South East Asia, and a robust growth is seen over the decades indicating that the
potential of this palm to generate revenue for the country’s economic growth. A complete
understanding on the mechanisms of plant growth promoting activity by the beneficial bacteria
cultivated into the biofertilizers is important as this could further enhance the growth and
improve the yields, with less detrimental impacts on the agroecosystems for a more sustainable
plantation.
Objectives

1. To identify the potential plant growth promoting rhizobacteria in Innotech biofertilizer
2. To study the effects of biofertilizers on the growth of oil palm seeds in different type

of soils.
3. To determine the effects of biofertilizers on the soil properties and nutrient uptake in

different type of soils.

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2.0 METHOLODOGY
Isolation and identification of bacteria
Given the bacterial content in Innotech biofertilizer was unknown, a 16S rRNA sequencing
approach was done to identify the most dominant bacterial strains in order fully understand
their functions and applications in promoting the oil palm seedlings growth. The 16S ribosomal
RNA sequences (only for bacteria) obtained from NCBI were then compared with the existed
sequence databases in BLAST and phylogenetic trees was established using Neighbour-joining
bootstrap analysis with 1000 replications.
Glasshouse study
The experiment was conducted at University of Malaya glasshouse in Rimba Ilmu where four
different types of soils (peat soil, topsoil, clay soil and red gravel soil) used in this experiment
to study the effects on growth of oil palm (Plate 1). The soils were provided by Innotech Bio
Sdn Bhd, and the oil palms nursery seedlings used were Felda Yangambi (Plate 2). Four gram
of Christmas Island Phosphate Rock (CIRP) was applied and mixed into each of the polybags
and incubated for a week before the transplant. Two treatments were designed with 4 replicates
are listed in Table 1.

Plate 1 : Four different types of soil
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Plate 2: Preparation of glasshouse study

Table 1: List of treatments Descriptions
Treatments 100% of chemical fertilizers
Treatment 1 70% of chemical fertilizers + 30% of
Treatment 2 Innotech Fertilizer

• 4 replicates

Measurement of the plant growth, nutrient uptake and soil analysis.

During the treatment period, the growth parameters of the oil palms were observed and taken
every 2 weeks. The amount of chlorophyll was measured using a SPAD chlorophyll meter at
the 3rd frond from the left. The height of the seedlings was measured using the 5th frond from
the left and taken from the lowest rudimentary to the tip of the rachis. The girth size was
measured using a digital Vernier calliper at 5 cm from the planting medium.

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After 1-month planting

The oil palms were harvested after 4 months of planting. They were carefully removed from
the soil and the roots were cleansed of soil particles. The oil palms were cut at the soil level
and separated from the roots. The dry mass of both shoots and root were determined by drying
in an oven at 71-75 ºC until constant weight was achieved. The shoots and roots were ground
using a grinding machine (<2 mm) separately for macronutrients (NPK) analysis. The soil used
was thoroughly mixed and air dried, sieved using 2 mm mesh sieve and measured for
macronutrients (NPK) content. Nitrogen content was analysed using TruMac CNS, while
phosphorus and potassium were analysed using Auto-Analyser. The results were expressed as
mean ± standard deviation.

Statistical analysis
Tukey's HSD (honest significant difference) test will be used to determine significant
differences (p< 0.05) between different types of soil orders and t-Test will be used to determine
significant differences (p< 0.05) between two treatments. The data are statistically analyzed
using the IBM SPSS Statistics for Windows, Version 20.0

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3.0 RESULTS AND DISCUSSION
3.1 Isolation and identification of bacteria
The identification of the three most dominant bacterial strains (named UM-A, UM-B, and UM-
E) using 16S rRNA sequencing followed by BLAST to establish the phylogenetic trees
depicted that Bacillus spp were the dominant species in Innotech biofertilizer. The identified
Bacillus strains were all gram positive; UM-A as Bacillus tequilensis strain 10b, UM-B as
Bacillus amyloliquefaciens strain MPA 1034 and UM-E as Bacillus cereus strain JCM
2152 (fig. 2-4). These Bacillus strains have been reported in few published papers as potential
PGPRs.

Figure 2: UM-A phylogenetic tree (Neighbour-joining bootstrap analysis with 1000
replications)

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Figure 3: UM-B phylogenetic tree (Neighbour-joining bootstrap analysis with 1000
replications)

Figure 4: UM-E phylogenetic tree (Neighbour-joining bootstrap analysis with 1000
replications)
Three dominant strains of bacteria were identified in the liquid culture of Innotech biofertilizer,
namely Bacillus tequilensis strain 10b, Bacillus amyloliquefaciens strain MPA 1034 and
Bacillus cereus strain JCM 2152. These strains of PGPR have been reported to enhance
growth and fix nitrogen with several non-leguminous crops (Amir et al., 2004). Bacillus group
of bacteria have been listed as one of the N2-fixing bacteria associated with the non-legumes
nitrogen fixation (Wani, 1990).
Bacillus strains contained in Innotech biofertilizer have been reported to enhance plant growth.
B. tequilensis was reported by (Dastager, Deepa et al. 2011) to produce indole-3-acetic acid
(IAA) which promoted the proliferation and elongation of the black pepper roots. The strain
used by Dastager et al. is a phosphate solubilizer and it improved the macronutrient (NPK)
uptake of the black pepper in both acidic and alkaline conditions.
On the other hand, B. amyloliquefaciens is efficient in producing secondary metabolites such
as lipopeptides, surfactins, bacillomycin D, and fengycins, which are secondary metabolites
mainly with inhibiting pathogens activity (Chen, Vater et al. 2006, Almaghrabi, Massoud et al.
2013) B. amyloliquefaciens SAHA 12.07 which has 99% of similarity with B.

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amyloliquefaciens strain MPA 1034 was also reported to inhibit Ganoderma boninense through
the production of chitinases which degrade the chitin walls of most pathogenic fungi (Azizah,
Mubarik et al. 2015).
B. amyloliquefaciens was also able to produce mixture of organic acids such as lactic,
isovaleric, isobutyric and acetic acid that were identified as phosphate solubilizers (Illmer,
Schinner et al. 1992).
A more recent study by (Yanti, Warnita et al. 2018) reported that Bacillus cereus strain JCM
2152 has the potential to promote the growth of the tomato plant and provide resistance towards
Ralstonia solanacearum which usually causes bacterial wilt in tomato and other tropical crops.
Thus, the Bacillus content in Innotech biofertilizer would be a good bioformulation to enhance
and protect the crops from various diseases.

3.2 Plant Growth
Over the 4 months period (December 2017 – end of March 2018) for 131 days, the growth
attributes of the oil palm seedlings were monitored, and the results showed a positive response
towards the combined treatment between NPK fertilizer with Innotech biofertilizer compared
to the treatment with only NPK fertilizer.

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a) Height

The observation on height of the highest fronds depicted that seedlings under T2 especially in
peat and topsoil have the longest fronds, compared to seedlings under T1 using the same type
of soils. Seedlings in peat, topsoil and clay seemed to have better growth performance under
the inoculation of Innotech biofertilizers except for seedlings in gravel soil. Initially at 67 DAT,
seedlings in topsoil were higher than the rest of the seedlings planted in other soils, but peat
soils generally shown better growth at the end of the treatment period. In terms of the height
between both treatments, seedlings in topsoil under T2 shown a very significant increase with
the inoculation of biofertilizer up to 14% increment from T1. As compared to seedlings in peat
soil under T2, an increment of 3% from T1 was observed even though seedlings under this
condition have the highest fronds height among other soils.

140.00
120.00

Height, cm 100.00 Peat
80.00 Topsoil
60.00 Clay soil
40.00 Gravel soil
20.00

0.00 T2
T1

T1 T2

125.00 130.00
115.00
105.00 120.00

Height, cm 95.00 Peat Height, cm 110.00 Peat
85.00 Topsoil 100.00 Topsoil
75.00 Claysoil Claysoil
65.00 Gravel soil 90.00 Gravel soil
80.00
67 131
67 131 Days after transplant (DAT)
Days after transplant (DAT)

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b) 5th frond height

A different trend was observed in the height of the fifth frond where seedlings in topsoil and
clay soil under treatment with Innotech biofertilizer have the highest height of their fifth frond.
A 13% increase in fifth frond height was seen in seedlings planted using topsoil under T2 in
comparison with T1 seedlings. Seedlings in peat soil treated with chemical fertilizers, however
shown a significant fifth frond height, compared to the other type of soil within the same
treatment and T2. But, in general all seedlings in T2 shown a noticeable trend of increment
starting 41 DAT except for seedlings in gravel soil which have constant (very slight) change in
their fifth frond height from 41 DAT. With the inoculation of biofertilizer, seedlings in peat
and topsoil particularly performed better in their highest frond and fifth frond height. This
proves the efficiency of beneficial microbes in promoting the growth of the oil palm seedlings.

5th frond height 80.00 Peat soil
70.00 Top soil
60.00 Clay soil
50.00 Gravel soil
40.00
30.00 Peat
20.00 Topsoil
10.00 Clay soil
Gravel soil
0.00

T1 T2

T1 T2

70.00 65.00
63.00
5th Frond height, cm 65.00 5th frond height, cm 61.00
59.00
60.00 Peat 57.00
Topsoil 55.00
55.00 Claysoil 53.00
Gravel soil
50.00 30 41 67 131
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45.00
30 41 67 131

c) Girth size
Earlier results of the highest frond height and fifth frond height were supported by the
observation made on the girth size of the seedlings. The line graph demonstrates that all
seedlings in T2 increased in their girth size 41 DAT as compared to seedlings in T1. The graph
also suggested that the girth size of seedlings in topsoil under the inoculation with Innotech
biofertilizer was the biggest, followed by peat soil and topsoil. Seedlings in gravel soil showed
poor girth enhancement in both treatments. Although seedlings in topsoil generally have the
biggest girth among the seedlings in other soils, a significant increment of 18% was observed
in seedlings in peat soil treated with Innotech biofertilizer. Seedlings treated with biofertilizers
have shown an enhanced girth size as early as 41 DAT, compared to seedlings treated with
chemical fertilizer. Seedlings in T1 have a rather slowed enhancement in its girth size (starting
from 67 DAT).

Girth size, mm 45.00 Peat
40.00 Top soil
35.00 Clay soil
30.00 Gravel soil
25.00
20.00 T2
15.00
10.00

5.00
0.00

T1

T1 T2

40.00 45.00
40.00
Girth size, mm 35.00 Girth size, mm 35.00
30.00
30.00 Peat 25.00 Peat
25.00 Topsoil 20.00 Topsoil
20.00 Claysoil 15.00 Claysoil
15.00 Gravel soil Gravel soil

30 41 67 131 30 41 67 131
Days after transplant (DAT) Days after transplant (DAT).

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d) Chlorophyll
The difference between chlorophyll content of seedlings in T1 and T2 is very significant. T1
seedlings, however shown a declining trend especially in topsoil, clay and gravel soil with
seedlings in peat depicted a gradual increase in its chlorophyll content. Through the treatment
with Innotech biofertilizer, the chlorophyll content of seedlings in peat, topsoil and clay soil
significantly increased since 30 DAT, except for seedlings planted in gravel soil that fluctuated
at 41 DAT. A very subtle increase in chlorophyll was seen in seedlings in topsoil and clay from
30 DAT, but seedlings under peat shown a very high chlorophyll content as of 41 DAT
compared to the other types of soil under the treatment with Innotech biofertilizer. Increments
of 5% and 8% respectively were seen in seedlings planted in peat and topsoil under T2 when
compared to the chlorophyll production in T1 using the same soils. Chlorophyll is essential to
the photosynthesis of the seedlings, and this enhanced production of chlorophyll is due to the
efficiency of beneficial microbes in fixing nitrogen (to the form which can be taken up by the
seedlings), and this nitrogen is responsible for the cellular synthesis of chlorophyll (Hayat et
al., 2010).

64.00

62.00

60.00

Chlorophyll 58.00 Peat
Topsoil
56.00 Clay soil
Gravel soil
54.00

52.00

50.00

T1 T2

T1 T2

Chlorophyll 60.00 Peat Chlorophyll 64.00 30 41 67 131 Peat
59.00 Topsoil 62.00 Topsoil
58.00 Claysoil 60.00 Claysoil
57.00 Gravel soil 58.00 Gravel soil
56.00 56.00
55.00 54.00
54.00 52.00
53.00 50.00
52.00
51.00
50.00

30 41 67 131

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e) Number of fronds
The inoculation of Innotech biofertilizer have no significant effect on the number of fronds,
but seedlings in gravel soil particularly have slightly more fronds compared to the other types
of soil in both treatments, followed by seedlings planted in peat soil. A 5% increase in fronds
production was seen in seedlings treated with the biofertilizer under gravel soil. Although, the
growth of seedlings in terms of the height and size of girth planted using gravel soil was poor,
but the seedlings expressed its growth by producing more fronds.

Number of fronds 16.00 Peat soil
14.00 Topsoil
12.00 Clay soil
10.00 Gravel soil

8.00 T2
6.00
4.00
2.00
0.00

T1

T1 T2

No. of frond 14.00 Peat No. of frond 15.00 Peat
13.00 Topsoil 14.00 Topsoil
12.00 Claysoil 13.00 Claysoil
11.00 Gravel soil 12.00 Gravel soil
10.00 11.00
10.00
9.00
8.00 9.00
7.00 8.00
6.00 7.00
6.00
30 41 67 131
30 41 67 131

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f) Shoot and root dry mass
The highest shoot dry mass was obtained from oil palms seedlings treated under T2 while the
least dry mass was from seedlings planted under T1 condition. This showed that the combined
treatment with Innotech biofertilizer have profound effect on the dry mass. As seen in Figure
5, larger root networks can be observed in seedlings under peat and topsoil as compared to clay
and gravel soil. However, plant growth analysis shown that seedlings in topsoil have good
growth enhancement in T2, but the shoot dry mass results signified the lowest reading. The
roots development of seedlings in peat and topsoil was much better than seedlings in clay and
gravel soil due to its low bulk density with increasing pore space and aeration and this helped
in enhancing the nutrient uptake by the seedlings, to promote growth.

Furthermore, poor root development was seen in clay and gravel soil. This can be caused by
the soil bulk density, with less pore space and soil aeration (Rosenani et al., 2016) which gave
less space for the roots proliferation. Seedlings treated under Innotech biofertilizer seemed to
have a positive impact on the root development and proliferation. This indicates the
effectiveness of PGPR to colonize the roots and promote nutrients solubilization and uptake by
the seedlings that result in enhanced growth.

Ratio of root-to-shoot showed that seedlings in peat and topsoil under treatment with the
biofertilizer is lower than seedlings under the treatment with chemical fertilizer. Roots weight
of seedlings in peat soil with biofertilizer were generally lower but they have higher shoot
weight. This can be explained through their optimization towards the available resources,
where in nutrient sufficient condition plants allocate relatively less to their roots as less effort
is required to acquire this resource ( Agren et al., 2003). However, the allocation of root and
shoot was not affected by the light availability as this experiment was conducted in a
randomized complete block design (RCBD). Nonetheless, it is important to have a better
understanding on how plants uptake and regulate the available nutrients in order to obtain a
more quantitative root:shoot allocation (Agren et al., 2003)

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Table 2 : Effects of different treatments and types of soil on the shoot and root dry mass, and root:shoot

Peat Soil T1 Shoot (g) Root (g) Root:shoot
Top soil T2
Clay soil T1 57.97 16.22 0.28
Gravel soil T2 62.07 16.10 0.26
T1 49.62 14.25 0.29
T2 63.48 16.52 0.26
T1 53.61 11.70 0.22
T2 66.34 15.20 0.23
65.97 15.55 0.24
58.70 14.30 0.25

Peat soil Top soil Clay soil Gravel soil

T1

T2

Figure 5
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3.3 Soil Macronutrients content
Table 3: The effects of different treatments and types of soil on soil macronutrient NPK

SOIL TREATMENT N (%) Macronutrients K (mg/L)
PEAT 0.61 ± 0.05 P (mg/L) 45.52 ± 0.23
TOPSOIL T1 0.61 ± 0.03 5.83 ± 0.19 46.15 ± 0.74
CLAY T2 0.34 ± 0.01 8.95 ± 0.95 48.79 ± 4.09
GRAVEL T1 0.35 ± 0.03 5.24 ± 2.11 51.54 ± 6.67
T2 0.10 ± 0.00 5.43 ± 1.81 40.19 ± 4.60
T1 0.11 ± 0.00 2.71 ± 0.11 49.94 ± 1.37
T2 0.12 ± 0.01 4.00 ± 0.52 47.11 ± 1.78
T1 0.14 ± 0.02 2.08 ± 0.98 55.03 ± 2.79
T2 3.01 ± 0.62

Values are mean of n = 3 for each type of treatment. Data was expressed as mean ± standard deviation. Means
sharing the same letter among the treatments do not differ significantly at (P < 0.05).

Accumulatively, soils treated under T2 have higher macronutrient (NPK) content compared to
T1 as shown in Table 3. A significant difference was observed between T1 and T2 on the N
content in the clay soil. However, there was a variability of N content in all soils, with clay and
gravel soils have the lowest N %. This could be due to the uptake of N by the seedlings, in
which N was responsible for cellular synthesis of chlorophyll and other components for the
plant growth (Hayat et al., 2010). Phosphorus content in the soil with treatment T2 was higher
than T1 especially in peat soil. This suggests that PGPR in commercial Innotech Fertilizer
exerted better phosphate solubilization abilities. The low P content in T1 seedlings could be
due to the poor phosphate solubilization potential such as Bacillus strains.

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3.4 Root and Shoot Macronutrients Content

Table 4: The effects of different treatments and types of soil on soil macronutrient NPK

SOIL TREATMENT SHOOT ROOT

N (mg/L) P (mg/L) K (mg/L) N (mg/L) P (mg/L) K (mg/L)

PEAT T1 47.07 ± 3.20 2.78 ± 0.27 33.66 ± 2.14 38.60 ± 3.32 1.41 ± 0.09 33.81 ± 1.64

T2 42.90 ± 1.37 2.83 ± 0.17 32.72 ± 1.22 37.87 ± 3.91 1.55 ± 0.12 32.94 ± 2.02

TOPSOIL T1 35.57 ± 2.28 3.10 ± 0.17 31.57 ± 1.37 30.47 ± 2.41 1.41 ± 0.21 32.99 ± 0.26

T2 36.57 ± 2.79 2.81 ± 0.50 32.13 ± 0.98 32.13 ± 2.17 1.31 ± 0.26 33.58 ± 0.99

CLAY T1 43.33 ± 5.85 3.69 ± 1.08 33.62 ± 2.35 40.93 ± 4.68 1.64 ± 0.27 33.23 ± 1.45

T2 40.23 ± 1.42 3.22 ± 0.27 34.85 ± 4.36 36.27 ± 2.59 1.54 ± 0.13 32.67 ± 1.70

GRAVEL T1 33.70 ± 3.50 3.56 ± 0.34 33.37 ± 0.86 30.77 ± 1.42 1.33 ± 0.35 32.99 ± 0.41

T2 36.70 ± 3.70a 3.90 ± 0.51 32.16 ± 0.48 32.50 ± 3.72 1.59 ± 0.26 34.02 ± 2.84

Values are mean of n = 3 for each type of treatment. Data was expressed as mean ± standard deviation. Means
sharing the same letter among the treatments do not differ significantly at ( P < 0.05).

In general, the nitrogen content in the shoot of the seedlings planted using peat soil under T1
was the highest. However, the combined treatments with biofertilizers have no effect on the
nitrogen uptake in all seedlings. But, the treatments depicted a profound effect on the nitrogen
uptake in shoot and roots of all seedlings under topsoil and gravel soil. However, there were
no significant differences observed between all treatments.

The phosphorus content in shoot of seedlings under T2 was the highest in gravel soil. This
result was consistent with the uptake of phosphorus by the roots. Shoots of seedlings in peat
soil, however, have the lowest phosphorus level compared to the other soils, but seedlings in
T2 had an increment by 1.7% from T1.

The uptake of potassium by the shoots of the seedlings was almost the same across all the soils
and treatments. T2 seedlings under clay soil, have the highest uptake of potassium. Although
the nutrients uptake by seedlings was higher in T1 but there were no significant differences
observed between all treatments.

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4.0 CONCLUSION
In this study, we were able to demonstrate the effectiveness of biofertilizers in promoting the
growth of seedlings in the nursery stage. Three dominant strains of bacteria were identified in
the liquid culture of Innotech biofertilizer, namely Bacillus tequilensis strain 10b, Bacillus
amyloliquefaciens strain MPA 1034 and Bacillus cereus strain JCM 2152. Therefore, Innotech
biofertilizer has the potential to increase various plant growth parameters of the oil palm
seedlings. The combined treatment of biofertilizers from Innotech with addition of chemical
fertilizer seems to have a profound effect on the enhancement of the oil palms growth and
appears to be the best alternative to the sole application of NPK fertilizers. From this study,
peat and topsoil have a great potential to be used in oil palm nurseries to enhance the growth
of the seedlings before being transplanted into the real planting sites. Seedlings in topsoil under
T2 (with Innotech biofertilizer) depicted an impressive total of 14%, 13% and 8% increment
than T1 (without Innotech biofertilizer) in height, 5th frond height and the chlorophyll
respectively. While peat soil shown increment of 18% in girth size of its seedlings, 3% and 5%
in height and chlorophyll respectively. The application of Innotech biofertilizer improved the
nutrients uptake and increased the girth size of the seedlings as compared to the sole treatment
with compound fertilizers. These Bacillus strains were ideal to be incorporated into
biofertilizer formulation as they will be able to promote the plant growth, provided that a further
research is needed to observe the bacteria efficiency in protecting the crops from diseases and
harsh environmental conditions. Further work includes transplanting the seedlings into real
plantation field to monitor the effectiveness of biofertilizer in larger scale is recommended.

5.0 REFERENCES
Almaghrabi, O. A., Massoud, S. I., and Abdelmoneim, T. S. (2013). Influence of inoculation

with plant growth promoting rhizobacteria (PGPR) on tomato plant growth and nematode
reproduction under greenhouse conditions. Saudi Journal of Biological Sciences. 20. 57-
61.

Amir, H. G., Shamsuddin, Z. H., Halimi, M. S., Marziah, M., and Ramlan, M. F. (2004).
Enhancement in nutrient accumulation and growth of oil palm seedlings caused by
PGPR under field nursery conditions. Communications in Soil Science and Plant
Analysis, 36 (15-16), 2059-2066.

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Agren, G. I., and Franklin, O. (2003). Root:Shoot Ratios, Optimization & Nitrogen
Productivity. Annals of Botany. 92(6). 795-800.

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Acknowledgement
The author greatly acknowledges the financial support from Innotech Bio. Sdn. Bhd.

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6.0 APPENDICES

Figure 6: The difference between seedlings under T1 (left) and T2 (right) in Topsoil. The
seedling in T2 have an enhanced growth compared to the seedling in T1.

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