Fadil

Journal of Tropical Crop Science Vol. 6 No.
3, October 2019
www.j-tropical-crops.com
Humic Acid and Biofertilizer Applications Enhanced Pod

and Cocoa Bean Production during the Dry Season at
Kaliwining Plantation, Jember, East Java, Indonesia
Fadil RohmanA, Ade WachjarB*, Edi SantosaB, and Soetanto AbdoellahC

A
Graduate School, IPB University, Bogor 16680, West Java, Indonesia.
B
Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor 16680,
West Java, Indonesia.
C
Indonesian Coffee and Cocoa Research Institute, Jember 68152, East Java, Indonesia.
*Corresponding author; email: wachjarade@yahoo.co.id
Abstract challenges related to poor soil fertility, pest and

disease attacks and climate change effects (ICCRI,
Cocoa (Theobroma cacao L.) is an important crop 2015; Santosa et al., 2018). From 2012 to 2016,
in Indonesia, but many farmers still face problem cocoa bean production in Indonesia decreased to
in improving bean production. This research aimed 658,399 tons indicating a 11.09% decline grown in
to evaluate the effect of humic acid and biofertilizer an area of 1.72 million hectares indicating a 3.03%
applications on pod growth and yield of cocoa. The decrease in area under production (Ditjenbun, 2017).
research was conducted at Kaliwining Plantation
managed by Indonesian Coffee and Cocoa Research In Indonesia, cocoa cultivation generally relies on
Institute at Jember District, East Java, Indonesia from NPK fertilizers from inorganic sources; although
June 2017 to February 2018. The experiment used cocoa intercropping to optimize fertilizer input is also
mature tree of Sulawesi from one clone. Treatment common (Santosa et al., 2005). Santosa et al. (2018)
used were humic acid at level of 0, 1000, 2000, 3000 speculated that high rainfall will have a detrimental
and 4000 ppm in combination with biofertilizer at effect on cocoa plantation because of high nutrient
level of 0, 500, 1000, 1500 and 2000 ppm that were leaching and disease outbreak. Therefore, there is
applied through soil and foliar respectively. Results of a need to improve fertilizer management in relation
this experiment showed that there was an interaction to climate change in order to conserve soil nutrients
between humic acid and biofertilizer on beans number in the plantation. Many researchers have reported
per plant and photosynthesis rate. Plants treated with that the use of inorganic fertilizers in a long term
1000 ppm humic acid produced the highest number has negative impacts on soil structure, cause water
of small cherelle. Biofertilizer applied at 1500 ppm pollution and disturb soil micro-organism (Massah
increased cherelle number, healthy cherelle, number and Azedagan, 2016; Zhou et al., 2017).
of young pods, number of harvested pods, bean
weight per plant and bean yield. The combination Recently, the application of organic fertilizers such as
of 1000 ppm humic acid and 1500 ppm biofertilizer humic acid and biofertilizer is getting popular in the
increased bean production by 39.7%. The high bean plantation. Humic acid is an active substance derived
production was in line with the high photosynthetic from organic matter that has functional group of –
rate. Thus, humic acid and biofertilizer applications COOH, phenolic –OH, carboxylic group and –C=O
could be a way to increase cocoa bean production in (Darmokoesoemo et al., 2014). Humic acid could
this area. improve soil permeability, aggregation, water holding
capacity, cation exchange capacity, form complex
Keywords: Theobroma cacao, cherelle wilt, climate compounds with heavy metals and increases
change, cocoa bean, photosynthesis. nitrogen, phosphorus, potasium and sulfur availability
(Tan, 2009; Hermanto et al., 2012). Humic acid could
stimulate plant growth (Dariah and Nurida, 2011), and
Introduction according to Suwardi and Wijaya (2013), increase
in plant growth after humic acid treatment is due to
Cocoa (Theobroma cacao L.) is an important crop enhancement activities of endogenous hormones
in Indonesia, but production sustainability faces such as auxin, gibberellin and cytokinin. However,
Humic Acid and Biofertilizer Applications Enhanced Pod and Cocoa Bean .......... 153
Journal of Tropical Crop Science Vol. 6 No. 3, October 2019
organic fertilization application in cocoa plantation lamtoro (Leucaena leucocephala) in block 1 and 2,
using humic acid has rarely been studied. while block 3 used mixed of lamtoro and teak trees
(Tectona grandis). Population of L.leucocephal was
Biofertilizer application stimulates plant growth 555 plants for all blocks, while T. grandis was 84
and production through mechanisms of phosphate plants per hectare along the edge of the block 3.
mineral dilution, nitrogen fixation and phytohormones
production (Ramakrishnan and Selvakumar, 2012; Humic acid (Humatop®, Indonesia) according to
Siagian et al., 2014; Khan et al., 2016; Sahu et al., the manufacturer’s description was from leonardite
2017; Setyadi et al., 2017). Siagian et al. (2014) rocks with solubility of 99%, pH 8.6, 38.81%
stated that biofertilizers contain microorganisms C-organic content, 10.31% K2O, 6,503 ppm Fe, 20
such as Azospirillum sp., Bacillus sp., Pseudomonas ppm Mn, 23 ppm Zn and water content of 16.56%.
sp. and Trichoderma sp; and Azospirillum sp. which Biofertilizer (Bactoplus ®, Indonesia) according to the
could fix nitrogen air to stimulate plant growth. The manufacturer’s description contained Azospirillum
existence of Bacillus sp. and Pseudomonas sp. sp. 2.35 x 106, Bacillus endophiticus 1.12 x 107,
according to Sivasakthi et al. (2014), was able to Pseudomonas flourescens 8.40 x 106 and Tricoderma
synthesize IAA, dissolve phosphate and increase N, sp. 1.50 x 105 cfu.g-1. Both materials were purchased
P, and K absorption. Pratama et al. (2013) reported from the commercial stores.
that microorganisms in biofertilizers could also
improve plant resistance to Phytophtora palmivora, Treatments
an important disease in cocoa plantation. The study
aimed to evaluate the effect of humic acid and Research was conducted using complete randomized
biofertilizer applications on cocoa pod growth and block design with two factors. The first factor was
bean production. humic acid concentration, i.e., 0, 1000, 2000, 3000
and 4000 ppm obtained by dissolving 0, 1, 2, 3 and
4 g.L-1 of powder in water, respectively. The second
Methods factor was biofertilizer, i.e., 0, 500, 1000, 1500 and
2000 ppm obtained by dissolving i.e. 0, 0.5, 1.0, 1.5
Site and Materials and 2.0 g. L-1 of powder in water, respectively. Each
combination was repeated three times, so there
Research was conducted from June 2017 to February were 75 experimental units. Each experimental unit
2018 at Kaliwining Plantation, a farm managed by consists of 16 trees, and 4 trees at the middle of block
Indonesian Coffee and Cocoa Research Institute was selected as samples.
(ICCRI), Jember District, East Java, Indonesia.
The peak of the rainy season at the study site was Humic acid was applied in the soil at 75 cm from
November to May, thus the research was conducted the stem while the biofertilizer was sprayed onto
predominantly during the dry season. The climatic the canopy. The control treatment used was water.
data during the course of the research were as follows: Treatment application was conducted three times
average monthly rainfall was 129 mm, average daily after every four weeks starting on the 19th of June
minimum temperature (T) was 20.95 °C and maximum 2017. The application of inorganic fertilization (N,
was 32.51 °C, air relative humidity was 89.98%, wind P, K, Mg) was based on the Indonesian Coffee and
velocity was 0.53 km hour-1, sunshine duration was Cacao Research Institute (ICCRI) standard, where
65.21%, and evaporation was 3.27 mm. Soil had annually a tree received urea (46% N) 220 g, TSP
pH (H2O) 4.27 and CEC 15 cmol.kg-1 with C-organic (36% P2O5) 180 g, KCl (60% K2O) 170 g and Kieserit
content, N, P, K, Ca, Mg and Na were 3.79%, 0.26%, (60% MgO) 115 g, which were applied one third each
147.36 ppm, 1.46 cmol.kg-1, 12.74 cmol kg-1, 8.69 in January, May and November. Pest and disease
cmo. kg-1 and 0.95 cmol. kg-1, respectively. were controlled using Nordox 86 WG ® and Azure
200 EC ® at 500 L. ha-1 of each, applied in November.
Experiment was conducted on one clone of cocoa
from Sulawesi aged 5−15 years with 3−5 m in height. Observation and Measurement
Treatment was arranged in blocks that represented
replications. The blocks had different trees aged Observations and measurements were made on
5, 10 and 15 years, represented as block 1, 2 and pods in the main stem and branches located 75 cm
3, respectively. Cocoa planting space was 3 x 3 m from jorquette for 24 weeks after treatment (WAT)
using the standard population of 1,111 plants per at 2-week intervals (Figure 1). Number of harvested
hectare. Plants were derived from top grafting using pods, number of beans per plant, single bean weight,
scion plagiotropic branch, height of jorquette was bean weight per plant and productivity data were
10−60 cm from the soil surface. Shade tree used was observed from 24 to 28 WAT in a weekly interval.
154 Fadil Rohman, Ade Wachjar, Edi Santosa, and Soetanto Abdoellah
Figure 1. Ten-year-old ocoa trees in the field (A); shade area for cherelles and pods measurement (B).
Drawings are not to scale.
Pods were classified based on their length into tiny- all cherelle developed into pod as some cherelles
sized cherelle (TC) <5cm, small-sized cherelle (SC) wilted. Number of TC was affected by biofertilizer,
5−10 cm, medium-sized pod (MP) 10−15 cm and while number of SC was affected by single treatment
large-sized pod (LP) >15 cm. Photosynthetic rate, of humic acid and biofertilizer. On average, TC
transpiration rate, stomatal conductance, Ci/Ca developed into SC was 55% and SC developed
ratio and water use efficiency (WUE) were observed into medium pod then large pod was 62%. The
using LICOR 6400 XT at 24 WAT. Pod growth and percentage of survived TC and SC were not affected
production data were analyzed using ANCOVA, while by humic acid but affected by biofertilizer application.
physiological data were analyzed using ANOVA. The higher concentration of biofertilizer produced
Significant means were further analyzed using more number of healthy cherelles.
Duncan’s Multiple Range Test (DMRT) at α=5% using
SAS software 9.4. Humic acid application on cocoa tree had no significant
effect on total TC at 24 WAT (Table 1). On the other
hand, biofertilizer spraying significantly affected
Results total TC and SC, percentage of healthy TC and SC.
Interaction between humic acid and biofertilizer had
Plant Growth no effect on total and percentage of healthy cherelles.
Plant growth could be observed visually from flush Plants treated with 1000 ppm biofertilizer significantly
emergence and the intensity of shoot formation. produced higher total TC than control and 500 ppm,
Cocoa tree treated with humic acid and biofertilizer but were not significantly different with 1500 and 2000
had more flush than those trees that did not receive ppm biofertilizers (Table 2). At 4−24 WAT, the highest
any treatments. The high number of flushes was total TC were obtained after application of 2000 ppm
followed by new shoot (chupons) growth that were biofertilizer (Figure 2). Plants sprayed with 1500
also higher than those of the control. Compared to the biofertilizer significantly produced higher number
control, the number of chupons in the treated plants of healthy TC than the other treatments (Table 2).
were approximately 34% higher, but the presence of Application level of 1500 ppm biofertilizer consistently
the new shoot were not observed further. Extensive showed the highest percentage of healthy TC at
new shoot was detrimental on to pod development, 12−24 WAT (Figure 2).
therefore, during tree maintenance the undesirable
shoots were removed. At 24 WAT, total SC of plants treated with 2000 ppm
humic acid was significantly higher than the other
Tiny-sized (TC) and Small-sized Cherelle (SC) treatments. Biofertilizer application at the same level
produced SC 43% higher than the control (Table
Cocoa tree produces flowers and cherelles throughout 2). The combination of humic acid and biofertilizer
the year and they do not depend on tree age, as at 2,000 ppm each showed the most favor for SC
observed in all blocks. Total cherelles per observation production at 8−24 WAT (Figure 3). Nevertheless,
reached 2.4−6.4 and 1.4−4.4, consecutively for tiny percentage of healthy SC did not corelate to the
cherelle (TC) and small cherelle (SC). However, not number of SC especially on the application of humic
acid (Table 1). Percentage of healthy SC of plants
Table 1. Summary of covariance and variance analysis of pod growth and physiological variables at 24 week
after treatment (WAT) and harvest variables at 28 WAT
Variable Humic Acid Biofertilizer Interaction
Total tiny cherelle (TC) 0.99 ns 3.39 * 1.42 ns
Healthy TC (%) 1.14 ns 9.61** 0.56 ns
Total small cherelle (SC) 5.53 ** 4.86 ** 0.81 ns
Healthy SC (%) 0.70 ns 2.91 * 1.81 ns
Medium-sized pod (MP) 0.45 ns 10.81 ** 1.00 ns
Large-sized pod (LP) 0.43 ns 1.60 ns 0.66 ns
Harvested pods number 0.53 ns 3.86 ** 1.74 ns
Beans number per plant 0.81 ns 7.65 ** 1.98 *
Single bean weight 0.33 ns 2.47 ns 0.81 ns
Beans weight per plant 0.74 ns 5.94 ** 1.82 ns
Yield per harvest 0.80 ns 5.71 ** 1.72 ns
Photosynthetic rate 1.41 ns 26.38 ** 1.87 *
Transpiration rate 0.38 ns 0.36 ns 1.37 ns
Stomatal conductance 0.74 ns 0.40 ns 0.88 ns
Ci/Ca 0.61 ns 17.83 ** 1.45 ns
WUE 1.27 ns 19.81** 1.37 ns
Note: * = P<0.05, ** = P<0.01 and ns = not significant according to Fisher test α 5%; where Ci/Ca = ratio of CO2 intercellular
to CO2 atmosphere; WUE = water use efficiency.
treated with 1500 ppm biofertilizer was significantly Medium-sized (MP) and Large-sized Pod (LP), and
higher than the control and the biofertilizer applied at Bean Production
500 ppm (Table 2). The highest percentage of healthy
SC at 12−24 WAT was produced by plants treated Number of medium-sized pod (MP) was not affected
with 1500 ppm biofertilizer (Figure 3). by humic acid application (Table 1). Spraying of 1500
ppm biofertilizer significantly increased MP number
Table 2. Number of tiny-sized cherelles (TC), small-sized cherelle (SC), medium-sized pod (MP) and large-
sized pod (LP) on humic acid and biofertilizer treatments at 24 WAT
Treatment Number of TC Number of SC Number of pod
(ppm) Total Healthy (%) Total Healthy (%) MP LP
Humic acid
0 (control) 5.3 a 41.23 a 2.7 c 58.82 a 1.8 a 1.2 a
1000 5.8 a 41.30 a 4.2 a 53.43 a 1.9 a 1.3 a
2000 5.4 a 42.49 a 3.5 b 56.13 a 2.0 a 1.3 a
3000 5.6 a 43.62 a 3.5 b 56.75 a 2.0 a 1.3 a
4000 5.5 a 46.32 a 3.3 bc 59.97 a 2.0 a 1.2 a
Biofertilizer
0 (control) 5.1 b 37.57 b 2.8 c 50.61 b 1.6 b 1.2 a
500 5.0 b 38.42 b 3.0 bc 52.93 b 1.7 b 1.2 a
1000 5.8 a 42.31 b 3.5 ab 59.16 ab 1.9 a 1.2 a
1500 5.7 a 53.92 a 3.9 a 64.42 a 2.4 a 1.4 a
2000 5.8 a 42.71 b 4.0 a 57.92 ab 2.0 a 1.3 a
Note: Values followed by different letters within a column are significantly different according to DMRT α=5%. Interactions
among treatments are shown in Table 1.
Figure 2. Total and percentage of tiny-sized cherelle (TC) on different level of humic acid (A-B) and biofertilizer
applications (C-D) at different weeks after treatment (WAT); ns = not significant according to Fisher
test α = 5%.
Figure 3. Total and percentage of small-sized cherelle (SC) of different level of humic acid (A-B) and biofertilizer
applications (C-D) at 2 to 24 weeks after treatment (WAT); ns = not significant according to Fisher
test α = 5%.
in 24 WAT as compared to the control and 500 ppm the higher level of biofertilizer stimulated higher
(Table 2). Plants treated with 1500 ppm biofertilizer photosynthesis rate. However, photosynthesis rate
produced the highest MP at 12−24 WAT (Figure 4). was relatively constant, irrespective of humic acid
Figure 4. Number of medium-sized pods (MP) at different level of humic acid (A) and biofertilizer application
(B) at 2 to 24 weeks after treatment (WAT); ns = not significant according to Fisher test α = 5%).
Application of 1500 ppm biofertilizer showed the levels, i.e. 41.2−44.7 µmol CO2 m-2.s-1.
highest number of harvested pods, cocoa bean weight
per plant and yield per harvest at 27 and 28 WAT. Humic acid application had no effect on Ci/Ca ratio of
Increasing number of harvested pods, bean weight cocoa tree at 24 WAT (Table 1). On the other hand,
per plant and yield per plant at 28 WAT were 32.8%, application of 1,500 ppm biofertilizer significantly
39.7% and 39.4%, respectively, as compared to the decreased Ci/Ca ratio compared to the control, 500
control (Figure 5). Figure 5C showed that single bean and 1000 ppm, but it was not significantly different with
weight was relatively constant among harvesting 2000 ppm (Table 5). Table 5 shows that transpiration
times and among treatments, i.e., 1.04−1.10 g. rate and stomatal conductance were not affected by
fertilizer treatment. Water use efficiency (WUE) was
Interaction of humic acid and biofertilizer significantly 50-54%, irrespective of humic acid treatment at 24
affected number of beans per plant at 28 WAT (Table WAT, while increasing level of biofertilizer higher
1). Cocoa tree that received a combination of humic than 500 ppm significantly increased WUE. Spraying
acid and biofertilizer at 1000 ppm each produced the biofertilizer at the rate of 1500 ppm significantly
higherst number of cocoa beans per plant (Table 3). increased WUE of cacao plants by 17−41% as
compared to 0−1000 ppm (Table 5).
Photosynthesis
Discussion
The interaction of humic acid and biofertilizer
significantly affected photosynthesis rate in 24 WAT The application of biofertilizer increased cocoa
(Table 1). The combination of 3000 ppm humic production (Table 2), and at 1000−2000 ppm
acid and 1500 ppm biofertilizer showed the highest significantly resulted in a higher bean weight per
photosynthesis rate (Table 4). Table 4 shows that plant and yield relative to the control. The increase in
Figure 5. Number of harvested pod (A), beans weight per plant (B), individual bean weight (C) and yield
per harvest (D) at different level of biofertilizer at 24 to 28 weeks after treatment (WAT); where * =
significant different according to DMRT α 5%.
Table 3. Number of beans per plant as affected by different level of humic acid and biofertilizer treatments at
28 weeks after treatment (WAT)
Biofertilizer Humic acid level (ppm)
level (ppm) Average
0 1000 2000 3000 4000
0 17.3 ef 23.1 cdef 24.8 cdef 22.1 cdef 25.3 cdef 22.5
500 22.8 cdef 16.5 f 22.4 cdef 26.0 cdef 25.7 cdef 22.7
1000 28.5 abcd 37.9 a 26.4 bcdef 20.7 def 24.4 cdef 27.6
1500 29.9 abcd 29.2 abcd 27.8 abcde 37.4 ab 32.3 abc 31.3
2000 28.7 abcd 28.7 abcd 23.7 cdef 31.5 abcd 30.6 abcd 28.6
Average 25.4 27.1 25.0 27.5 27.7 26.5
Note: Values followed by different letters within a column and a row are significantly different according to DMRT α=5%.
Table 4. Photosynthetic rate (µmol CO2 m-2.s-1) of cocoa tree with different level of humic acid and biofertiliz-
er treatments at 24 WAT
Biofertilizer Humic acid level (ppm)
level (ppm) Average
0 1000 2000 3000 4000
0 36.12 fgh 33.72 h 33.27 h 39.09 defgh 39.09 defgh 36.3
500 37.28 efgh 33.24 h 34.39 gh 41.13 cdefgh 40.21 defgh 37.3
1000 41.28 cdefgh 50.26 abc 43.26 cdefg 36.73 efgh 44.86 bcdef 43.3
1500 48.16 abcd 53.72 ab 47.42 abcd 56.03 a 48.55 abcd 50.8
2000 46.06 bcde 48.55 abcd 47.56 abcd 50.31 abc 43.43 cdefg 47.2
Average 41.8 43.9 41.2 44.7 43.2 42.9
Note: Values followed by different letters within a column and a row are significantly different according to DMRT α=5%.
Table 5. Transpiration rate, stomatal conductance, Ci/Ca ratio and water use efficiency (WUE) as affected by
different levels of humic acid and biofertilizer treatments at 24 WAT
Fertilizer level Transpiration rate(m- Stomatal conductance (mol
Ci/Ca WUE
(ppm) mol H2O m-2.s-1) H2O m-2.s-1)
Humic acid
0 8.42 a 0.30 a 0.33 a 0.50 a
1000 8.22 a 0.29 a 0.30 a 0.54 a
2000 8.31 a 0.30 a 0.34 a 0.50 a
3000 8.28 a 0.30 a 0.30 a 0.54 a
4000 8.33 a 0.29 a 0.31 a 0.52 a
Biofertilizer
0 8.33 a 0.30 a 0.42 a 0.44 c
500 8.41 a 0.30 a 0.41 a 0.45 c
1000 8.29 a 0.29 a 0.31 b 0.53 b
1500 8.30 a 0.30 a 0.19 c 0.62 a
2000 8.23 a 0.30 a 0.25 bc 0.58 a
Note: Values followed by different letters within a column are significantly different according to DMRT α=5%. Interaction
among treatments are shown in Table 1.
production is likely due to an increase in the number and yield of cacao beans. The application of 1000
of harvested pods and bean weight per plant. It is ppm humic acid and 1500 ppm biofertilizer increased
also noted that spraying with 1500 ppm biofertilizer cocoa bean production by 39.7%. Based on this
increases the number of total cherelles and the results application of humic acid and biofertilizer is
percentage of healthy cherelle (Table 2). The high recommended to increase cocoa production in the
percentage of healthy cherelle means high chance to dry season.
obtain harvested pods. Khattab et al. (2012) stated
that application of humic acid on pomegranate plants
increased fruit number per tree and production by Acknowledgement
10.8% and 24.6%, respectively. In apple, Hidayatullah
et al. (2018) revealed that the application of humic The authors thank the Director and members of the
acid increases fruit number by 470%. staff of the Indonesian Coffee and Cocoa Research
Institute (ICCRI), Jember District, East Java,
Increasing bean weight per plant on the tree that Indonesia for supporting the research.
received biofertilizer spraying is likely related to an
increase in the photosynthetic rate (Tabel 4); higher
photosynthesis means higher photosynthate that References
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Journal of Tropical Crop Science Vol. 6 No.

Humic Acid and Biofertilizer Applications Enhanced Pod

Fadil RohmanA, Ade WachjarB*, Edi SantosaB, and Soetanto AbdoellahC

*Corresponding author; email: wachjarade@yahoo.co.id

Abstract challenges related to poor soil fertility, pest and

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