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Int. J. Agri. Agri. R.
Marin
Page 1
RESEARCH PAPER OPEN ACCESS
Jatropha-based alley cropping system’s contribution to carbon
sequestration
Rico A. Marin*
College of Forestry and Environmental Science, Central Mindanao University, Musuan, Bukidnon,
Philippines
Article published on January 12, 2016
Key words: Alley cropping, Agroforestry, Carbon sequestration, Jatropha curcas, hedgerow.
Abstract
The study was conducted to evaluate the total carbon stocks sequestered in a Jatropha – based alley cropping
system treated with varying fertilizer applications. The study was laid out in Randomized Complete Block Design
with three replications. The alley was planted with corn in two seasons Treatments include control (no fertilizer),
organic fertilizer and inorganic fertilizer applied to the alley crops. Findings showed that the treatments with
fertilizer applications had higher carbon stock in the jatropha hedges. The carbon content of the corn stover was
also higher in organic and inorganic fertilizer-applied treatments. However, highest soil carbon content was
shown in treatments applied with organic fertilizer (4.28 Ton ha-1). The inorganic fertilizer treatment had the
lowest soil carbon content with a mean of 4.28 Ton ha-1. In terms of total carbon stock of the entire jatropha-
based alley cropping system, there was a significant difference among treatments with organic fertilizer
application having the highest mean of 7.79 Ton ha-1 while the inorganic treated plots had 6.53 Ton ha-1. The no
fertilizer treatment had the least carbon stocks with 6.53 Ton ha-1. This recent study revealed that the jatropha-
based alley cropping system is a potential land-use for carbon sequestration. This farming system needs to be
promoted in upland areas to function not only as soil and water conservation measures but also as a possible
remedy for global warming.
* Corresponding Author: Rico A. Marin  ricomarin@yahoo.com
International Journal of Agronomy and Agricultural Research (IJAAR)
ISSN: 2223-7054 (Print) 2225-3610 (Online)
http://www.innspub.net
Vol. 8, No. 1, p. 1-9, 2016
Int. J. Agri. Agri. R.
Marin
Page 2
Introduction
One of the approaches recognized as a suitable
farming technology for upland areas is agroforestry
(Lasco and Visco, 2003). This technology was
believed to play a major role in halting and even
reversing the decline in agricultural production
(Chivenge, 2006). Agroforestry has also an important
role in providing livelihood and restoring the
watershed and environmental functions of the
uplands (PCARRD, 2006).
The alley cropping system of agroforestry is
commonly adopted in sloping lands (Palada et al. as
cited by Matta-Machado et al., 1995). This involves
planting of hedgerows along the contours and
growing agricultural crops in the alleys formed
between the hedgerows (Lasco and Visco, 2003).
Hedgerow plants can be trees or other plants that
have multiple uses, which can provide a range of
benefits to farmers, and have the capacity to retard
soil loss.
Jatropha curcas (Physic Nut or Jatropha) plant,
having been recognized as a possible alternative
source of biodiesel, has been grown all throughout the
world. The Philippine government is becoming
interested on the massive production of this plant as a
potential solution to the current petroleum crises. The
areas identified for jatropha plantation in the
Philippines are marginal sites or other sites not used
for prime agricultural crop production. These areas
are usually located in sloping lands or in moderately
rugged terrains. The said areas are in most cases also
very suitable for agroforestry practices like alley
cropping. In fact, there are already existing alley
cropping systems in the Philippines using J. curcas as
hedgerows.
Land uses such as an agroforestry system have a
significant role in moderating climate change since
they can be sources and sinks of carbon (Lasco et al.,
2001). The potential to sequester C in agroforestry
systems in tropical and temperate regions is
promising, but little information is available to date
(Oelbermann et al., 2004).
This study that investigated the extent of carbon
sequestration of jatropha-based alley cropping
system, therefore, is of interest. This attempted to
quantify the contribution of jatropha-based alley
farming system in mitigating climate change by
determining its extent in carbon sequestration.
Materials and methods
Location and Description of the Study Site
The study was conducted at the 2-year old Jatropha
based alley cropping system of Central Mindanao
University, Musuan, Bukidnon (Fig. 1). The average
elevation of the site is 430 m asl and the slope
gradient ranges from 25 to 30 %.
Fig. 1. The study site.
Experimental Design and Lay-out
The study was conducted for two croppings. It was
laid-out in Randomized Complete Block Design
(RCBD) replicated three times (Appendix 1). The
following treatments were as follows:
Treatment 1 Control (No Fertilizer)
Treatment 2 Organic Fertilizer
Treatment 3 Inorganic Fertilizer
Int. J. Agri. Agri. R.
Marin
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Plot length per treatment was sixteen meters (16 m)
with half a meter buffer at the edge of the alley and
one (1) meter buffer per treatment. Distance between
rows of the alley was 0.67m. X-Tekh Bio-fertilizer was
used in this study as the biofertilizer treatment.
Application was done using a knapsack sprayer and
the prescription was based on the standard
recommendation or protocol of the company. The
application protocols were as follows (XTekh Flier,
nd):
10 – 15 DAP : 35 ml or 50 ml X-Tekh Micro/ 16L
water
25 – 30 DAP : 70 ml or 50 ml X-Tekh Micro/ 16L
water
35 – 45 DAP : 100 ml X-Tekh Micro/ 16L water. Add
100 ml X-Tekh Potassium/16 liter
Two 2 bags of fertilizer (complete)/hectare were
applied per hectare apart from the liquid X-tekh bio-
fertilizer.
Inorganic fertilizers applied were based on the
recommendations from the analysis of the soil
samples by the Soils and Plant Analysis Laboratory of
the CMU - College of Agriculture.
Cultural Treatments
Weeding was done in the experimental area as the
need arises. Prior to the cropping period, hedgerows
were pruned at a distance of 50 cms from the ground
and the pruning were scattered uniformly on the alley
to serve as mulch.
Data collection
Hedgerow (Jatropha curcas):
Destructive sampling was done to assess the carbon
stocks of the hedgerow plants. Samples of J. curcas
hedgerow were harvested in each treatment.
Harvested samples were weighed using a scale and
biomass values were determined using the formula
(Pulhin, 2009):
Where: ODW = Total oven dry weight
TFW = total Fresh weight
SFW = Sample Fresh weight
SODW = Sample oven-dry weight
The below ground biomass (roots) was determined
using the formula (Pulhin, 2009):
Root Biomass = Exp [-1.0587 + 0.8836 ×LN(AGB)]
Where: Exp = e to the of
LN = natural log
AGB = Above Ground Biomass
Carbon estimates were determined using the formula
(Pulhin, 2009):
Carbon in tree biomass = Biomass × % Carbon in Tree
Alley crop (Maize)
Within the 16 meter length per treatment, four 1 x 1
meter square quadrat were established. All stover of
mature (harvestable) age within the quadrats were
taken for biomass determination. The formula in
determining the biomass and carbon stocks for the
alley crop were the same with the method used in the
hedgerow.
Soil Organic Carbon
Within the established quadrats, soil samples were
collected for soil carbon determination. Soil samples
were obtained within the 0-30 cm depth. The soil
samples were brought to the CMU College of
Agriculture Soil and Plant Analysis and Laboratory
(SPAL) for soil organic matter and bulk density
determination. Soil samples were collected thrice for
determination of carbon sequestration in each
treatment. One was done before the start of the first
cropping, another before the start of the second
cropping and the third collection was before the
harvest of the second cropping.
After the soil organic matter was obtained, the soil
organic carbon (SOC) was computed using the
formula below (De La Cruz, undated):
Int. J. Agri. Agri. R.
Marin
Page 4
In order to express the amount of SOC from percent
to tons C/ha, the following formula were used (De La
Cruz, undated):
 Weight of soil/ha = 10,000m2 × 0.30 m × bulk
density ,
 Ton C in soil/ha = % SOC × wt of soil in kg/ha × 1
ton/1000kg .
Total Carbon Sequestration:
The total carbon sequestration per treatment of the
Jatropha based alleycropping system was determined
using the formula:
Total carbon = hedgerow + alley crop + soil
Data Analysis
The test of significant difference among treatments
was determined using the Analysis of Variance
(ANOVA). Least significant difference and Tukey’s, on
the other hand, were used in comparing treatment
means. The Statistical Package for the Social Sciences
(SPSS) version 16 was used in the data analyses.
Results and discussion
Table 1 presents the biomass of the Jatropha curcas
hedgerow during the first and second cropping of the
said agroforestry system. Statistical analyses show no
significant difference among fertilizer treatments in
all the parameters measured both for the upper and
lower hedges.
Table 1. Hedge Biomass of the Jatropha based Alley cropping system.
Fertilizer Treatments First Cropping
(Ton Ha -1)
Second Cropping
(Ton Ha -1)
Biomass gain
(Ton Ha-1)
AGB BGB TGB AGB BGB TGB AGB BGB TGB
Upper Hedge
Control (F1) 2.37 0.74 3.11 2.49 0.78 3.27 0.13 0.03 0.16
Organic Fertilizer (F2) 2.72 0.84 3.56 2.88 0.88 3.76 0.16 0.05 0.20
Inorganic Fertilizer (F3) 2.58 0.80 3.38 2.74 0.84 3.58 0.15 0.05 0.20
Level of Significance ns ns ns ns ns ns ns ns ns
CV (%) 24.01 21.46 23.39 24.24 23.62 23.62 30.03 0.0 29.16
Lower Hedge
Control (F1) 2.76 0.85 3.61 2.90 0.89 3.79 0.13 0.037 0.173
Organic Fertilizer (F2) 2.81 0.86 3.67 2.95 0.90 3.85 0.14 0.040 0.177
Inorganic Fertilizer (F3) 3.04 0.93 3.97 3.19 0.97 4.16 0.15 0.043 0.197
Level of Significance ns ns ns ns ns ns ns ns ns
CV (%) 9.67 8.03 9.31 9.68 8.44 9.41 0.0 10.19% 0.0
Legend:
AGB: Above Ground Biomass
BGB: Below Ground Biomass
TGB: Total Ground Biomass
In the upper hedge, the biomass of the organic
fertilizer treatments had the highest values both for
the first and the second cropping. Total above ground
biomass of organic fertilizer treatment were 3.56 Ton
Ha-1 and 3.76 Ton Ha-1 for the first and second
cropping, respectively. The no fertilizer treatment had
the lowest with 3.11 Ton Ha-1 for the first cropping
and 3.27 Ton Ha-1 in the second cropping. In terms of
total biomass gain, both organic and inorganic
fertilizer treatments had 0.20 Ton Ha-1 while the no
fertilizer treatment had only 0.16 Ton Ha-1. The data
show that hedges where alley plots were applied with
fertilizers (organic and inorganic) are bigger in sizes
compared with hedges where alley plots have no
application with fertilizers. Hedgerows in this study
were not applied with fertilizer. The nutrients
received basically by the hedge plants came from
what were available in the soil and from those that
were leached out from the alley plots. It was further
believed that the nutrients leached out from the alley
Int. J. Agri. Agri. R.
Marin
Page 5
plots were partly trapped by the hedges and were
used up by J. curcas for its growth and development.
Hedgerow system can improve the capture and
cycling of nutrients (Wojtkowski, 2002). Nutrient loss
can be associated with soil erosion occurring in the
area. This is supported by Nelson et al. (1998a) who
pointed out that erosion differentially moves the
finer, most fertile soil fractions of the top soil.
The lower hedge shows that the biomass was found
highest in the inorganic fertilizer treatment. Data
show that the total biomass of the inorganic fertilizer
treatment was 3.97 Ton Ha-1 in the first cropping and
4.16 Ton Ha-1 in the second cropping. The treatment
with the least biomass was the no fertilizer with only
3.61 Ton Ha-1 and 3.79 Ton Ha-1 for the first and
second cropping, respectively. For the biomass gain at
the lower hedge, the inorganic fertilizer treatment had
the highest with 0.197 Ton Ha-1 and is closely followed
by organic fertilizer treatment with 0.177 Ton Ha-1.
The least among the treatments was the no fertilizer
treatment with only 0.173 Ton Ha-1. J. curcas in F3
and F2 plots were bigger than F1 which had the
shortest hedgerow plants among the treatments. The
relatively bigger sizes of F2 and F3 hedges can be
attributed to the fertilizer applied in the alley crops. It
is again presumed that some of the fertilizer applied
to the alley crops also benefited the hedgerow plants.
The hedge carbon of the jatropha-based alley
cropping system showed no significant difference in
all parameters among the fertilizer treatments (Table
2). Same as with biomass, the organic fertilizer
treatments had the greatest total carbon in the upper
hedge with 1.60 Ton Ha-1 and 1.69 Ton Ha-1 for the
first and second cropping, respectively. The no
fertilizer treatment had the lowest with only 1.4 Ton
Ha-1 in the first cropping and 1.47 Ton Ha-1 during the
second cropping. On the other hand, the carbon gain
between the first and second cropping shows that
both organic and inorganic fertilizer treatments had
0.09 Ton Ha-1 as against no fertilizer treatment with
only 0.07 Ton Ha-1. The higher sizes of the F2 and F3
hedges which resulted to higher biomass is basically
the reason why carbon stocks were high in these
treatments. Tandug et al. (2010) stressed that
diameter and total height of trees are important
variables to biomass gain. Biomass is directly
proportional to the amount of carbon stored in plants.
Table 2. Hedge Carbon of the Jatropha-Based Alley cropping system.
Fertilizer Treatments First Cropping
(Ton Ha -1)
Second Cropping
(Ton Ha -1)
Carbon Gain
(Ton Ha -1)
AGC BGC TGC AGC BGC TGC AGC BGC TGC
Upper Hedge
Control 1.06 0.33 1.40 1.12 0.35 1.47 0.06 0.017 0.07
Organic Fertilizer 1.22 0.38 1.60 1.30 0.40 1.69 0.07 0.017 0.09
Inorganic Fertilizer 1.16 0.36 1.52 1.23 0.38 1.61 0.07 0.020 0.09
Level of Significance ns ns ns ns ns ns ns ns ns
CV (%) 24.0 21.58 23.54 24.22 22.21 23.60 0.0 29.62 37.46
Lower Hedge
Control (F1) 1.24 0.38 1.62 1.31 0.40 1.71 0.063 0.02 0.077
Organic Fertilizer (F2) 1.27 0.39 1.65 1.33 0.41 1.73 0.063 0.02 0.080
Inorganic Fertilizer (F3) 1.37 0.42 1.79 1.43 0.43 1.87 0.070 0.02 0.087
Level of Significance ns ns ns ns ns ns ns ns ns
CV (%) 9.48 7.98 9.55 9.90 7.65 9.28 13.46 0.0 11.98
Legend:
AGC: Above Ground Carbon
BGC: Below Ground Carbon
TGC: Total Ground Carbon
Int. J. Agri. Agri. R.
Marin
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In the lower hedge, inorganic fertilizer treatment had
the highest total carbon for both first and second
cropping with 1.79 Ton Ha-1 and 1.87 Ton Ha-1,
respectively. The least among the treatment in total
carbon was the no fertilizer treatments with 1.61 Ton
Ha -1 in the first cropping and 1.71 Ton Ha -1 during
the second cropping. In terms of carbon gain at the
lower hedge, inorganic fertilizer treatment was
highest with 0.087 Ton Ha -1 and was followed by
organic fertilizer with 0.080 Ton Ha -1 while the no
fertilizer treatment was lowest with 0.077 Ton Ha-1.
Significant difference was shown in the carbon
content of the stover in the first cropping and there is
high significant in the second cropping (Table 3).
Data showed that organic fertilizer had the highest
amount of carbon stored with 0.11 Ton Ha-1 and 0.08
Ton Ha-1 in the first and second cropping,
respectively. However, its difference from inorganic
fertilizer is not significant. The no fertilizer treatment
had the lowest carbon stock for both first and second
cropping with 0.07 Ton Ha-1 and 0.03 Ton Ha-1,
respectively. No fertilizer treatment is significantly
different to both organic and inorganic fertilizer
treatment in the first cropping and highly significant
in the second cropping. This recent findings is parallel
to the investigation made by Allmaras et al. (2004)
that nitrogen fertilization increased stover C by 20%.
Table 3. Carbon stock in the corn stover of the
jatropha-based alley cropping system.
Treatments First
Cropping
Second
Cropping
No Fertilizer 0.07b 0.03b
Organic Fertilizer 0.11a 0.08a
Inorganic Fertilizer 0.09a 0.09a
Level of Significance * **
CV (%) 10.14 11.52
Means followed by the same letter in a column are not
significantly different at 5%.
Carbon content in plants can be associated with its
biomass production. The greater the biomass, the
higher will be its carbon content. In this study, those
treatments applied with fertilizers have greater
biomass content and carbon storage. Nutrients
provided by F2 and F3 treatments attributed to the
result of this study. F2 received commercial inorganic
fertilizer supplemented with X-Tekh biofertilizer
while F3 was applied with ammonium phosphate and
urea. Degree of application of X-Tekh for F2 was
based on the recommendation of the company, while,
that for F3 was according to the suggestion made
after soil analysis of the study site. This is consistent
with the findings of Tisdale et al. (1990) where an
adequate supply of nitrogen was found to be
associated with vigorous vegetative growth and a dark
green color of the plant. On the other hand, a good
supply of phosphorus has been associated historically
with increased root growth. The study of Ohlrogee
and associates (no date) as cited by Tisdale et al.
(1990) revealed that when soluble phosphate and
ammonium nitrogen are supplied together in a band,
plant roots will proliferate extensively in the area of
the treated soil. Good root system leads to better
uptake of nutrients desired by the crop for growth and
development.
In the soil carbon of the jatropha based alley cropping
system, no significant difference was shown among
treatments before the study was conducted. However,
after the first cropping, significant difference was
shown. Furthermore, high significant was shown
among treatments after the second cropping (Table
4). Among the treatments, organic fertilizer had the
highest carbon content both after the first and second
cropping with 4.91 Ton Ha-1 and 4.28 Ton Ha-1,
respectively. The no fertilizer treatment was the
second highest with 3.12 after the first cropping and
2.98 after the second cropping. However, its
difference from inorganic fertilizer, which had the
lowest, is not significant. The information reported by
Triberti et al (2008) has shown parallel findings with
this recent study where from 1972 to 2000 in the
Southeast Po valley (Italy), Soil Organic Carbon
(SOC) stock increased at mean rates of 0.16, 0.18, and
0.26 t ha−1 year−1 with the incorporation of residues,
slurry and manure, corresponding to sequestration
efficiencies of 3.7, 3.8 and 8.1% of added C with these
various materials. The authors added that manure,
Int. J. Agri. Agri. R.
Marin
Page 7
thus, confirmed its efficacy in increasing both SOC
content and soil fertility on the long-term.
Table 4. Soil Carbon of the jatropha-based alley
cropping system.
Treatments
Before
first
cropping
Before
Second
Cropping
After
second
cropping
No Fertilizer 3.75 3.12b 2.98b
Organic Fertilizer 3.52 4.91a 4.28a
Inorganic Fertilizer 2.38 3.05b 2.96b
Level of Significance ns * **
CV (%) 24.22 36.36 7.31
Means followed by the same letter in a column are not
significantly different at 5%.
Findings can be attributed to the organic fertilizer
used in the study. Xtekh biofertilzer is a bio-fertilizer
microorganism and potassium bio-fertilizer. It is a
revolutionary high technology liquid fertilizer
(organic based) consisting of organic and microbial
elements. The composition of this fertilizer may have
enhanced the increase of carbon content in the soil.
Accordingly, some of the benefits that can be derived
from X-tekh bio-fertilizer include: (a) improved root
development and growth leading to increased
nutrient uptake; (b) increase in crop yields; (c)
increase in quality – color, taste and size of fruit; (d)
increased resistance from pest and diseases; (e) aids
uniform crop establishment; (f) restores the normal
soil microflora; (g) improves soil structure for
optimum water retention; (h) reduces fertilizer
requirement; (i) reduces leaching problems, (j) is
environment friendly; and, (k) easy to apply and
handle (X-Tekh Flier, nd).
Table 5. presents the carbon stocks of the alley
farming system in the first and second cropping.
Findings show that the above ground carbon stocks
did not show significant difference both for the two
cropping period. Result, however, revealed that the
below ground carbon stock show significant
difference in the first cropping while highly
significant difference was shown in the second
cropping. Organic fertilizer treatments had the
highest carbon stocks both in the first and second
cropping. The inorganic and no fertilizer treatments,
on the other hand, are not significantly different.
Table 5. Total carbon sequestered in the alley cropping system.
Treatments
First Cropping Second Cropping
AGC BGC TC AGC BGC TC
No Fertilizer 2.38 3.84b 6.22b 2.58 3.73b 6.31b
Organic Fertilizer 2.60 5.68a 8.27a 2.70 5.08a 7.79a
Inorganic Fertilizer 2.62 3.83b 6.45b 2.76 3.77b 6.53b
Level of Significance ns * * ns ** *
CV (%) 10.14 12.08 10.86 10.48 6.17 6.87
Means followed by the same letter in a column are not significantly different at 5%.
Legend:
AGB: Above Ground Carbon
BGB: Below Ground Carbon
TC: Total Carbon
On total carbon stocks, significant difference were
shown in the two cropping system. Organic fertilizer
treatment had the highest carbon stock at 8.27 Ton
Ha-1 and 7.79 Ton Ha-1 for the first and second
cropping, respectively. No fertilizer treatment had the
lowest with 6.22 in the first cropping while 6.31 in the
second cropping. Its difference with the inorganic
fertilizer, however, is not significant.
The high below ground carbon in the organic fertilizer
treatment was attributed to its high carbon stock in
the soil. The use of organic rich fertilizer such as X-
Int. J. Agri. Agri. R.
Marin
Page 8
Tekh may have increased the carbon content of the
soil. In fact, the use of organic fertilizer has been
promoted in the recent times because it is believed to
be environmentally friendly and is providing healthy
products to the consumer. It has also several
advantages such as: (1) promotion of soil aggregation,
improvement of soil color, friability, and prevention
of soil erosion; (2) increase water holding capacity in
sandy soils; (3) supplies essential nutrients and
materials or increases organic matter level; (4)
increases soil cation exchange capacity; and, (5)
improves biological activities and enhances the rapid
multiplication of fungi, bacteria, actinomycetes and
other soil organisms.
Yan and Gong (2010) reported that the use of organic
fertilizer increased SOC and soil fertility and
consequently resulted in a larger yield trend when
compared to a balanced chemical fertilizer. Their
model simulation and pot experimental results also
indicated that soils with higher SOC had a higher
root/shoot ratio such that the long-term use of
organic fertilizer not only directly increases SOC, but
indirectly contributes to carbon sequestration by
favoring root development.
Conclusion
In summary, it can be concluded that jatropha based
alley cropping system has the potential in
sequestering carbon which is one of the causes of
global warming. Findings show that a total of 6.45 to
6.53 ton ha-1 has been stock in the said farming
system during the assessment. The alley crop (corn)
had sequestered a range of 0.03 to 0.11 Ton ha-1 for
the two croppings. Those crops which were fertilized
had more carbon stocks than those those without
fertilizer applications. The hedges aligned with those
alley crops applied with fertilizers had higher carbon
stocks than those hedges planted along with alley
crops with no fertilizer application. It was also shown
that the soil carbon content is enhanced by the
application of organic fertilizers in the alley crop.
Significant increase of carbon had been observed in
soils where application of organic substances is
employed.
Acknowledgement
The authors wish to express their thanks to the
University Administration headed by Dr. Maria Luisa
R. Soilven, President, for allocating funds in the
conduct of this study. Our thanks are also due to the
University Research Office for facilitating all required
documents and flow of communications during the
implementation of this research endeavor. Special
thanks is extended to the Forest Resources
Development Division of the University Income
Generation Program for the labor services.
Appendix 1. Experimental lay-out of the main plot.
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Jatropha-based alley cropping system’s contribution to carbon sequestration

  • 1. Int. J. Agri. Agri. R. Marin Page 1 RESEARCH PAPER OPEN ACCESS Jatropha-based alley cropping system’s contribution to carbon sequestration Rico A. Marin* College of Forestry and Environmental Science, Central Mindanao University, Musuan, Bukidnon, Philippines Article published on January 12, 2016 Key words: Alley cropping, Agroforestry, Carbon sequestration, Jatropha curcas, hedgerow. Abstract The study was conducted to evaluate the total carbon stocks sequestered in a Jatropha – based alley cropping system treated with varying fertilizer applications. The study was laid out in Randomized Complete Block Design with three replications. The alley was planted with corn in two seasons Treatments include control (no fertilizer), organic fertilizer and inorganic fertilizer applied to the alley crops. Findings showed that the treatments with fertilizer applications had higher carbon stock in the jatropha hedges. The carbon content of the corn stover was also higher in organic and inorganic fertilizer-applied treatments. However, highest soil carbon content was shown in treatments applied with organic fertilizer (4.28 Ton ha-1). The inorganic fertilizer treatment had the lowest soil carbon content with a mean of 4.28 Ton ha-1. In terms of total carbon stock of the entire jatropha- based alley cropping system, there was a significant difference among treatments with organic fertilizer application having the highest mean of 7.79 Ton ha-1 while the inorganic treated plots had 6.53 Ton ha-1. The no fertilizer treatment had the least carbon stocks with 6.53 Ton ha-1. This recent study revealed that the jatropha- based alley cropping system is a potential land-use for carbon sequestration. This farming system needs to be promoted in upland areas to function not only as soil and water conservation measures but also as a possible remedy for global warming. * Corresponding Author: Rico A. Marin  ricomarin@yahoo.com International Journal of Agronomy and Agricultural Research (IJAAR) ISSN: 2223-7054 (Print) 2225-3610 (Online) http://www.innspub.net Vol. 8, No. 1, p. 1-9, 2016
  • 2. Int. J. Agri. Agri. R. Marin Page 2 Introduction One of the approaches recognized as a suitable farming technology for upland areas is agroforestry (Lasco and Visco, 2003). This technology was believed to play a major role in halting and even reversing the decline in agricultural production (Chivenge, 2006). Agroforestry has also an important role in providing livelihood and restoring the watershed and environmental functions of the uplands (PCARRD, 2006). The alley cropping system of agroforestry is commonly adopted in sloping lands (Palada et al. as cited by Matta-Machado et al., 1995). This involves planting of hedgerows along the contours and growing agricultural crops in the alleys formed between the hedgerows (Lasco and Visco, 2003). Hedgerow plants can be trees or other plants that have multiple uses, which can provide a range of benefits to farmers, and have the capacity to retard soil loss. Jatropha curcas (Physic Nut or Jatropha) plant, having been recognized as a possible alternative source of biodiesel, has been grown all throughout the world. The Philippine government is becoming interested on the massive production of this plant as a potential solution to the current petroleum crises. The areas identified for jatropha plantation in the Philippines are marginal sites or other sites not used for prime agricultural crop production. These areas are usually located in sloping lands or in moderately rugged terrains. The said areas are in most cases also very suitable for agroforestry practices like alley cropping. In fact, there are already existing alley cropping systems in the Philippines using J. curcas as hedgerows. Land uses such as an agroforestry system have a significant role in moderating climate change since they can be sources and sinks of carbon (Lasco et al., 2001). The potential to sequester C in agroforestry systems in tropical and temperate regions is promising, but little information is available to date (Oelbermann et al., 2004). This study that investigated the extent of carbon sequestration of jatropha-based alley cropping system, therefore, is of interest. This attempted to quantify the contribution of jatropha-based alley farming system in mitigating climate change by determining its extent in carbon sequestration. Materials and methods Location and Description of the Study Site The study was conducted at the 2-year old Jatropha based alley cropping system of Central Mindanao University, Musuan, Bukidnon (Fig. 1). The average elevation of the site is 430 m asl and the slope gradient ranges from 25 to 30 %. Fig. 1. The study site. Experimental Design and Lay-out The study was conducted for two croppings. It was laid-out in Randomized Complete Block Design (RCBD) replicated three times (Appendix 1). The following treatments were as follows: Treatment 1 Control (No Fertilizer) Treatment 2 Organic Fertilizer Treatment 3 Inorganic Fertilizer
  • 3. Int. J. Agri. Agri. R. Marin Page 3 Plot length per treatment was sixteen meters (16 m) with half a meter buffer at the edge of the alley and one (1) meter buffer per treatment. Distance between rows of the alley was 0.67m. X-Tekh Bio-fertilizer was used in this study as the biofertilizer treatment. Application was done using a knapsack sprayer and the prescription was based on the standard recommendation or protocol of the company. The application protocols were as follows (XTekh Flier, nd): 10 – 15 DAP : 35 ml or 50 ml X-Tekh Micro/ 16L water 25 – 30 DAP : 70 ml or 50 ml X-Tekh Micro/ 16L water 35 – 45 DAP : 100 ml X-Tekh Micro/ 16L water. Add 100 ml X-Tekh Potassium/16 liter Two 2 bags of fertilizer (complete)/hectare were applied per hectare apart from the liquid X-tekh bio- fertilizer. Inorganic fertilizers applied were based on the recommendations from the analysis of the soil samples by the Soils and Plant Analysis Laboratory of the CMU - College of Agriculture. Cultural Treatments Weeding was done in the experimental area as the need arises. Prior to the cropping period, hedgerows were pruned at a distance of 50 cms from the ground and the pruning were scattered uniformly on the alley to serve as mulch. Data collection Hedgerow (Jatropha curcas): Destructive sampling was done to assess the carbon stocks of the hedgerow plants. Samples of J. curcas hedgerow were harvested in each treatment. Harvested samples were weighed using a scale and biomass values were determined using the formula (Pulhin, 2009): Where: ODW = Total oven dry weight TFW = total Fresh weight SFW = Sample Fresh weight SODW = Sample oven-dry weight The below ground biomass (roots) was determined using the formula (Pulhin, 2009): Root Biomass = Exp [-1.0587 + 0.8836 ×LN(AGB)] Where: Exp = e to the of LN = natural log AGB = Above Ground Biomass Carbon estimates were determined using the formula (Pulhin, 2009): Carbon in tree biomass = Biomass × % Carbon in Tree Alley crop (Maize) Within the 16 meter length per treatment, four 1 x 1 meter square quadrat were established. All stover of mature (harvestable) age within the quadrats were taken for biomass determination. The formula in determining the biomass and carbon stocks for the alley crop were the same with the method used in the hedgerow. Soil Organic Carbon Within the established quadrats, soil samples were collected for soil carbon determination. Soil samples were obtained within the 0-30 cm depth. The soil samples were brought to the CMU College of Agriculture Soil and Plant Analysis and Laboratory (SPAL) for soil organic matter and bulk density determination. Soil samples were collected thrice for determination of carbon sequestration in each treatment. One was done before the start of the first cropping, another before the start of the second cropping and the third collection was before the harvest of the second cropping. After the soil organic matter was obtained, the soil organic carbon (SOC) was computed using the formula below (De La Cruz, undated):
  • 4. Int. J. Agri. Agri. R. Marin Page 4 In order to express the amount of SOC from percent to tons C/ha, the following formula were used (De La Cruz, undated):  Weight of soil/ha = 10,000m2 × 0.30 m × bulk density ,  Ton C in soil/ha = % SOC × wt of soil in kg/ha × 1 ton/1000kg . Total Carbon Sequestration: The total carbon sequestration per treatment of the Jatropha based alleycropping system was determined using the formula: Total carbon = hedgerow + alley crop + soil Data Analysis The test of significant difference among treatments was determined using the Analysis of Variance (ANOVA). Least significant difference and Tukey’s, on the other hand, were used in comparing treatment means. The Statistical Package for the Social Sciences (SPSS) version 16 was used in the data analyses. Results and discussion Table 1 presents the biomass of the Jatropha curcas hedgerow during the first and second cropping of the said agroforestry system. Statistical analyses show no significant difference among fertilizer treatments in all the parameters measured both for the upper and lower hedges. Table 1. Hedge Biomass of the Jatropha based Alley cropping system. Fertilizer Treatments First Cropping (Ton Ha -1) Second Cropping (Ton Ha -1) Biomass gain (Ton Ha-1) AGB BGB TGB AGB BGB TGB AGB BGB TGB Upper Hedge Control (F1) 2.37 0.74 3.11 2.49 0.78 3.27 0.13 0.03 0.16 Organic Fertilizer (F2) 2.72 0.84 3.56 2.88 0.88 3.76 0.16 0.05 0.20 Inorganic Fertilizer (F3) 2.58 0.80 3.38 2.74 0.84 3.58 0.15 0.05 0.20 Level of Significance ns ns ns ns ns ns ns ns ns CV (%) 24.01 21.46 23.39 24.24 23.62 23.62 30.03 0.0 29.16 Lower Hedge Control (F1) 2.76 0.85 3.61 2.90 0.89 3.79 0.13 0.037 0.173 Organic Fertilizer (F2) 2.81 0.86 3.67 2.95 0.90 3.85 0.14 0.040 0.177 Inorganic Fertilizer (F3) 3.04 0.93 3.97 3.19 0.97 4.16 0.15 0.043 0.197 Level of Significance ns ns ns ns ns ns ns ns ns CV (%) 9.67 8.03 9.31 9.68 8.44 9.41 0.0 10.19% 0.0 Legend: AGB: Above Ground Biomass BGB: Below Ground Biomass TGB: Total Ground Biomass In the upper hedge, the biomass of the organic fertilizer treatments had the highest values both for the first and the second cropping. Total above ground biomass of organic fertilizer treatment were 3.56 Ton Ha-1 and 3.76 Ton Ha-1 for the first and second cropping, respectively. The no fertilizer treatment had the lowest with 3.11 Ton Ha-1 for the first cropping and 3.27 Ton Ha-1 in the second cropping. In terms of total biomass gain, both organic and inorganic fertilizer treatments had 0.20 Ton Ha-1 while the no fertilizer treatment had only 0.16 Ton Ha-1. The data show that hedges where alley plots were applied with fertilizers (organic and inorganic) are bigger in sizes compared with hedges where alley plots have no application with fertilizers. Hedgerows in this study were not applied with fertilizer. The nutrients received basically by the hedge plants came from what were available in the soil and from those that were leached out from the alley plots. It was further believed that the nutrients leached out from the alley
  • 5. Int. J. Agri. Agri. R. Marin Page 5 plots were partly trapped by the hedges and were used up by J. curcas for its growth and development. Hedgerow system can improve the capture and cycling of nutrients (Wojtkowski, 2002). Nutrient loss can be associated with soil erosion occurring in the area. This is supported by Nelson et al. (1998a) who pointed out that erosion differentially moves the finer, most fertile soil fractions of the top soil. The lower hedge shows that the biomass was found highest in the inorganic fertilizer treatment. Data show that the total biomass of the inorganic fertilizer treatment was 3.97 Ton Ha-1 in the first cropping and 4.16 Ton Ha-1 in the second cropping. The treatment with the least biomass was the no fertilizer with only 3.61 Ton Ha-1 and 3.79 Ton Ha-1 for the first and second cropping, respectively. For the biomass gain at the lower hedge, the inorganic fertilizer treatment had the highest with 0.197 Ton Ha-1 and is closely followed by organic fertilizer treatment with 0.177 Ton Ha-1. The least among the treatments was the no fertilizer treatment with only 0.173 Ton Ha-1. J. curcas in F3 and F2 plots were bigger than F1 which had the shortest hedgerow plants among the treatments. The relatively bigger sizes of F2 and F3 hedges can be attributed to the fertilizer applied in the alley crops. It is again presumed that some of the fertilizer applied to the alley crops also benefited the hedgerow plants. The hedge carbon of the jatropha-based alley cropping system showed no significant difference in all parameters among the fertilizer treatments (Table 2). Same as with biomass, the organic fertilizer treatments had the greatest total carbon in the upper hedge with 1.60 Ton Ha-1 and 1.69 Ton Ha-1 for the first and second cropping, respectively. The no fertilizer treatment had the lowest with only 1.4 Ton Ha-1 in the first cropping and 1.47 Ton Ha-1 during the second cropping. On the other hand, the carbon gain between the first and second cropping shows that both organic and inorganic fertilizer treatments had 0.09 Ton Ha-1 as against no fertilizer treatment with only 0.07 Ton Ha-1. The higher sizes of the F2 and F3 hedges which resulted to higher biomass is basically the reason why carbon stocks were high in these treatments. Tandug et al. (2010) stressed that diameter and total height of trees are important variables to biomass gain. Biomass is directly proportional to the amount of carbon stored in plants. Table 2. Hedge Carbon of the Jatropha-Based Alley cropping system. Fertilizer Treatments First Cropping (Ton Ha -1) Second Cropping (Ton Ha -1) Carbon Gain (Ton Ha -1) AGC BGC TGC AGC BGC TGC AGC BGC TGC Upper Hedge Control 1.06 0.33 1.40 1.12 0.35 1.47 0.06 0.017 0.07 Organic Fertilizer 1.22 0.38 1.60 1.30 0.40 1.69 0.07 0.017 0.09 Inorganic Fertilizer 1.16 0.36 1.52 1.23 0.38 1.61 0.07 0.020 0.09 Level of Significance ns ns ns ns ns ns ns ns ns CV (%) 24.0 21.58 23.54 24.22 22.21 23.60 0.0 29.62 37.46 Lower Hedge Control (F1) 1.24 0.38 1.62 1.31 0.40 1.71 0.063 0.02 0.077 Organic Fertilizer (F2) 1.27 0.39 1.65 1.33 0.41 1.73 0.063 0.02 0.080 Inorganic Fertilizer (F3) 1.37 0.42 1.79 1.43 0.43 1.87 0.070 0.02 0.087 Level of Significance ns ns ns ns ns ns ns ns ns CV (%) 9.48 7.98 9.55 9.90 7.65 9.28 13.46 0.0 11.98 Legend: AGC: Above Ground Carbon BGC: Below Ground Carbon TGC: Total Ground Carbon
  • 6. Int. J. Agri. Agri. R. Marin Page 6 In the lower hedge, inorganic fertilizer treatment had the highest total carbon for both first and second cropping with 1.79 Ton Ha-1 and 1.87 Ton Ha-1, respectively. The least among the treatment in total carbon was the no fertilizer treatments with 1.61 Ton Ha -1 in the first cropping and 1.71 Ton Ha -1 during the second cropping. In terms of carbon gain at the lower hedge, inorganic fertilizer treatment was highest with 0.087 Ton Ha -1 and was followed by organic fertilizer with 0.080 Ton Ha -1 while the no fertilizer treatment was lowest with 0.077 Ton Ha-1. Significant difference was shown in the carbon content of the stover in the first cropping and there is high significant in the second cropping (Table 3). Data showed that organic fertilizer had the highest amount of carbon stored with 0.11 Ton Ha-1 and 0.08 Ton Ha-1 in the first and second cropping, respectively. However, its difference from inorganic fertilizer is not significant. The no fertilizer treatment had the lowest carbon stock for both first and second cropping with 0.07 Ton Ha-1 and 0.03 Ton Ha-1, respectively. No fertilizer treatment is significantly different to both organic and inorganic fertilizer treatment in the first cropping and highly significant in the second cropping. This recent findings is parallel to the investigation made by Allmaras et al. (2004) that nitrogen fertilization increased stover C by 20%. Table 3. Carbon stock in the corn stover of the jatropha-based alley cropping system. Treatments First Cropping Second Cropping No Fertilizer 0.07b 0.03b Organic Fertilizer 0.11a 0.08a Inorganic Fertilizer 0.09a 0.09a Level of Significance * ** CV (%) 10.14 11.52 Means followed by the same letter in a column are not significantly different at 5%. Carbon content in plants can be associated with its biomass production. The greater the biomass, the higher will be its carbon content. In this study, those treatments applied with fertilizers have greater biomass content and carbon storage. Nutrients provided by F2 and F3 treatments attributed to the result of this study. F2 received commercial inorganic fertilizer supplemented with X-Tekh biofertilizer while F3 was applied with ammonium phosphate and urea. Degree of application of X-Tekh for F2 was based on the recommendation of the company, while, that for F3 was according to the suggestion made after soil analysis of the study site. This is consistent with the findings of Tisdale et al. (1990) where an adequate supply of nitrogen was found to be associated with vigorous vegetative growth and a dark green color of the plant. On the other hand, a good supply of phosphorus has been associated historically with increased root growth. The study of Ohlrogee and associates (no date) as cited by Tisdale et al. (1990) revealed that when soluble phosphate and ammonium nitrogen are supplied together in a band, plant roots will proliferate extensively in the area of the treated soil. Good root system leads to better uptake of nutrients desired by the crop for growth and development. In the soil carbon of the jatropha based alley cropping system, no significant difference was shown among treatments before the study was conducted. However, after the first cropping, significant difference was shown. Furthermore, high significant was shown among treatments after the second cropping (Table 4). Among the treatments, organic fertilizer had the highest carbon content both after the first and second cropping with 4.91 Ton Ha-1 and 4.28 Ton Ha-1, respectively. The no fertilizer treatment was the second highest with 3.12 after the first cropping and 2.98 after the second cropping. However, its difference from inorganic fertilizer, which had the lowest, is not significant. The information reported by Triberti et al (2008) has shown parallel findings with this recent study where from 1972 to 2000 in the Southeast Po valley (Italy), Soil Organic Carbon (SOC) stock increased at mean rates of 0.16, 0.18, and 0.26 t ha−1 year−1 with the incorporation of residues, slurry and manure, corresponding to sequestration efficiencies of 3.7, 3.8 and 8.1% of added C with these various materials. The authors added that manure,
  • 7. Int. J. Agri. Agri. R. Marin Page 7 thus, confirmed its efficacy in increasing both SOC content and soil fertility on the long-term. Table 4. Soil Carbon of the jatropha-based alley cropping system. Treatments Before first cropping Before Second Cropping After second cropping No Fertilizer 3.75 3.12b 2.98b Organic Fertilizer 3.52 4.91a 4.28a Inorganic Fertilizer 2.38 3.05b 2.96b Level of Significance ns * ** CV (%) 24.22 36.36 7.31 Means followed by the same letter in a column are not significantly different at 5%. Findings can be attributed to the organic fertilizer used in the study. Xtekh biofertilzer is a bio-fertilizer microorganism and potassium bio-fertilizer. It is a revolutionary high technology liquid fertilizer (organic based) consisting of organic and microbial elements. The composition of this fertilizer may have enhanced the increase of carbon content in the soil. Accordingly, some of the benefits that can be derived from X-tekh bio-fertilizer include: (a) improved root development and growth leading to increased nutrient uptake; (b) increase in crop yields; (c) increase in quality – color, taste and size of fruit; (d) increased resistance from pest and diseases; (e) aids uniform crop establishment; (f) restores the normal soil microflora; (g) improves soil structure for optimum water retention; (h) reduces fertilizer requirement; (i) reduces leaching problems, (j) is environment friendly; and, (k) easy to apply and handle (X-Tekh Flier, nd). Table 5. presents the carbon stocks of the alley farming system in the first and second cropping. Findings show that the above ground carbon stocks did not show significant difference both for the two cropping period. Result, however, revealed that the below ground carbon stock show significant difference in the first cropping while highly significant difference was shown in the second cropping. Organic fertilizer treatments had the highest carbon stocks both in the first and second cropping. The inorganic and no fertilizer treatments, on the other hand, are not significantly different. Table 5. Total carbon sequestered in the alley cropping system. Treatments First Cropping Second Cropping AGC BGC TC AGC BGC TC No Fertilizer 2.38 3.84b 6.22b 2.58 3.73b 6.31b Organic Fertilizer 2.60 5.68a 8.27a 2.70 5.08a 7.79a Inorganic Fertilizer 2.62 3.83b 6.45b 2.76 3.77b 6.53b Level of Significance ns * * ns ** * CV (%) 10.14 12.08 10.86 10.48 6.17 6.87 Means followed by the same letter in a column are not significantly different at 5%. Legend: AGB: Above Ground Carbon BGB: Below Ground Carbon TC: Total Carbon On total carbon stocks, significant difference were shown in the two cropping system. Organic fertilizer treatment had the highest carbon stock at 8.27 Ton Ha-1 and 7.79 Ton Ha-1 for the first and second cropping, respectively. No fertilizer treatment had the lowest with 6.22 in the first cropping while 6.31 in the second cropping. Its difference with the inorganic fertilizer, however, is not significant. The high below ground carbon in the organic fertilizer treatment was attributed to its high carbon stock in the soil. The use of organic rich fertilizer such as X-
  • 8. Int. J. Agri. Agri. R. Marin Page 8 Tekh may have increased the carbon content of the soil. In fact, the use of organic fertilizer has been promoted in the recent times because it is believed to be environmentally friendly and is providing healthy products to the consumer. It has also several advantages such as: (1) promotion of soil aggregation, improvement of soil color, friability, and prevention of soil erosion; (2) increase water holding capacity in sandy soils; (3) supplies essential nutrients and materials or increases organic matter level; (4) increases soil cation exchange capacity; and, (5) improves biological activities and enhances the rapid multiplication of fungi, bacteria, actinomycetes and other soil organisms. Yan and Gong (2010) reported that the use of organic fertilizer increased SOC and soil fertility and consequently resulted in a larger yield trend when compared to a balanced chemical fertilizer. Their model simulation and pot experimental results also indicated that soils with higher SOC had a higher root/shoot ratio such that the long-term use of organic fertilizer not only directly increases SOC, but indirectly contributes to carbon sequestration by favoring root development. Conclusion In summary, it can be concluded that jatropha based alley cropping system has the potential in sequestering carbon which is one of the causes of global warming. Findings show that a total of 6.45 to 6.53 ton ha-1 has been stock in the said farming system during the assessment. The alley crop (corn) had sequestered a range of 0.03 to 0.11 Ton ha-1 for the two croppings. Those crops which were fertilized had more carbon stocks than those those without fertilizer applications. The hedges aligned with those alley crops applied with fertilizers had higher carbon stocks than those hedges planted along with alley crops with no fertilizer application. It was also shown that the soil carbon content is enhanced by the application of organic fertilizers in the alley crop. Significant increase of carbon had been observed in soils where application of organic substances is employed. Acknowledgement The authors wish to express their thanks to the University Administration headed by Dr. Maria Luisa R. Soilven, President, for allocating funds in the conduct of this study. Our thanks are also due to the University Research Office for facilitating all required documents and flow of communications during the implementation of this research endeavor. Special thanks is extended to the Forest Resources Development Division of the University Income Generation Program for the labor services. Appendix 1. Experimental lay-out of the main plot. References Allmaras RR, Linden DR, Clapp CE. 2004. Corn-Residue Transformations into Root and Soil Carbon as Related to Nitrogen, Tillage, and Stover Management. Soil Science Society of America Journal Vol 68, 1366-1375. Chivenge OA. 2006. Capacity building in Agroforestry in Africa and Southeast Asia. World Agroforestry into the future. World Agroforestry Center 135-140. Lasco RD, Sales RF, Extrella R, Saplaco SR, Castillo ASA, Cruz RVO, Pulhin FB. 2001. Carbon Stock Assessment of Two Agroforestry Systems in a Tropical Forest Reserve in the Philippines. The Philippine Agriculturist 84 (4), 401- 407.
  • 9. Int. J. Agri. Agri. R. Marin Page 9 Lasco RD, Visco RG. 2003. Introduction to agroforestry. A lecture Syllabus. Philippine Agroforestry Education and research network and UPLB Institute of Agroforestry 1-79. Matta-Machado RP, JORDAN CF. 1995. Nutrient dynamics during the first 3 years of an alley cropping agroecosystem in Southern USA. Agroforestry Systems: an International Journal Vol. 30(3), 351- 362. Nelson RA, Cramb RA, Menz K, Mamicpi M. 1998a. Gliricidia, napier, and natural vegetation hedgerows. In: K MENG, D MAGCALE- MACANDONG, and IW RUSASTRA (eds.). Improving small holder farming system in imperata areas of Southeast Asia: alternatives to shifting cultivation. Canberra, Australia:Aciar 95-112. Oelbermann M, Voroney RP, Gordon AM. 2004. Carbon sequestration in tropical and temperate agroforestry systems: a review with examples from Costa Rica and southern Canada. Agriculture, Ecosystems and Environment Vol. 104(3), 359-377. PCARRD. 1996. Alley cropping system in sloping lands. Technology Vol. 17(2). Los Baños, Laguna: PCARRD 1-2. PCARRD. 2006. The Philippine Recommends for Agroforestry. PCARRD Philippine Recommends Series no. 59-A. PCARRD; DOST; WAC (ICRAF) 1- 90. PCARRD-DOST. 2009. Compedium of information and Technologies on Jatropha . 1st Ed. Los Baňos Laguna: PCARRD_DOST 1- 86. Tandug LM, Umali MV Jr, Umali PA, Lantican NLM. 2010. Allometry of Aboveground Biomass and Carbon Content of Forest Tree Species. Canopy International 36, 1-6. Tisdale TS, Werner NL and Beaton JD. 1990. Soil fertility and fertilizers. Macmillan Publishing Company. New York. 4th edition 1 - 754. Triberti L, Nastri A,Giordani G, Comellini F, Baldoni G. 2008. European Journal of Agronomy. ELSEVIER. Volume 29, Issue 1,13–20. WAC. 2008. Introduction to agroforestry. World Agroforestry Center. http://www.worldagroforestry centre.org/Agrorestryuse.asp. Wojtkowski PA. 2002. Agroecological perspective in Agronomy, Forestry and Agroforestry. Science Publisher Inc 1-356. X-TEKH Bio-Fertilizer. The Nutrient Booster flier. Yang X, Gong W. 2010. The role of chemical and organic fertilizers on yield, yield variability and carbon sequestration-results of a 19-year experiment. Plant and Soil Vol. 331, Issue 1, 471-480.