JOURNAL OF DEGRADED AND MINING LANDS MANAGEMENT

Volume 10, Number 2 (January 2023):4255-4269, doi:10.15243/jdmlm.2023.102.4255

ISSN: 2339-076X (p); 2502-2458 (e), www.jdmlm.ub.ac.id

Research Article

The quality of acid soils treated with seaweed (Kappapychus alvarezii) sap
enriched biochar from Southeast Sulawesi, Indonesia
Laode Muhammad Harjoni Kilowasid1*, Dwita Siniona Manik1, Nevianti1, Gusti Arya
Komang1, Puja Mutmainna1, La Ode Afa1, Tresjia Corina Rakian1 , Waode Siti Anima
Hisein2, La Ode Ahmad Nur Ramadhan3, Syamsu Alam4
1 Department of Agrotechnology, Halu Oleo University, Kendari, 93232, Indonesia
2 Department of Plant Protection, Halu Oleo University, Kendari, 93232, Indonesia
3
Department of Chemistry, Halu Oleo University, Kendari, 93232, Indonesia
4
Department of Soil Science, Halu Oleo University, Kendari, 93232, Indonesia
*corresponding author: lohardjoni2@yahoo.co.id

Abstract
Article history: Biochar enriched with fertilizers has the potential to improve the quality of
Received 1 October 2022 acid soils. In the agricultural sector, it was reported that the sap of
Accepted 21 November 2022 Kappapychus alvarezii (K-sap) is often used as a biofertilizer. Therefore,
Published 1 January 2023 this study aimed to (i) examine the character of biochar enriched with the
K-sap and (ii) analyze the soil quality after incubation. This study was
carried out using two treatment factors arranged in a randomized block
Keywords: design, namely the biochar dose at 0%, 2.5%, 5.0%, and 7.5% in weight
acid soils and K-sap at concentrations of 0%, 5%, 10%, and 15%. Ten kilograms of
biochar acid soil were incubated with K-sap enriched biochar for up to 2 weeks in
functional group each experimental pot. The results showed that the absorption band
seaweed increased with the K-sap concentration for the functional group associated
soil fertility with the OH-stretching of the biochar enriched with K-sap was stronger
and wider than the unenriched. The peak elevation pattern detected by X-
ray diffraction for all biochars was similar. The dosage of 7.5% biochar
enriched with 5%-10% K-sap concentrations increased pH, EC, organic C,
total N, C:N ratio, available P, and exchangeable K, Ca, Mg, and Si than
those without treatment. Furthermore, ammonium and total spores of
arbuscular mycorrhizal fungi decreased, flagellates increased with and
without the biochar, while the nitrate, total bacteria, and fungi of all
treatments were similar. Based on these results, it was concluded that
biochar enriched with K-sap could improve the quality of acid soil.

To cite this article: Kilowasid, L.M.H., Manik, D.S., Nevianti, Komang, G.A., Mutmainna, P., Afa, L.O., Rakian, T.C. , Hisein,
W.S.A., Ramadhan, L.O.A.N. and Alam, S. 2023. The quality of acid soils treated with seaweed (Kappapychus alvarezii) sap
enriched biochar from Southeast Sulawesi, Indonesia. Journal of Degraded and Mining Lands Management 10(2):4255-4269,
doi:10.15243/jdmlm.2023.102.4255.

Introduction quality (Ibrahimi et al., 2022). Based on plant

The quality of soil plays an important role in achieving
sustainable crop productivity in agroecosystems
(Bünemann et al., 2018). Soils that are capable of
providing various ecosystem services such as fertility,
water storage, erosion control, carbon sequestration,
and food production are considered to be of good
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productivity, the quality level is reflected through
fertility, which shows the ability to provide nutrients
and the environmental conditions that ecologically
support plants' optimal growth and yields (Nguemezi
et al., 2020). Meanwhile, the level of availability of
nutrients in poor soils needed for plant growth is due
to the ecological interactions between the functional
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groups of soil biota such as bacteria, fungi,
mycorrhizal, and protozoa in a community of food
webs. Bacteria and fungi, including arbuscular
mycorrhizal, occupy positions at the basal trophic level
in food webs (Kilowasid et al., 2014). These organisms
recycle soil nutrients through the decomposition and
mineralization of the food web. Subsequently, the
decomposed nutrients available in the soil solution are
taken up by roots for plant growth (Salamon et al.,
2020). High soil acidity and low organic carbon
content significantly contribute to poor fertility and
beneficial biota in nutrient recycling in tropical
agroecosystems (Purwanto and Alam, 2020).
The emergence of acidification in tropical
agroecosystems is caused by an increase in H+ and Al3+
cations when Ca, Mg, K, and Na cations are leached
and replaced by H and Al cations in the adsorption area
(Aggegnehu et al., 2021). The addition of organic
matter also increases the pH and organic carbon of soil,
thereby improving the fertility of acid mineral soils
(Gafur and Umran, 2019). This organic matter must
have a long-term positive impact on high soil quality
for sustainable crop productivity (Krauss et al., 2020).
Moreover, the use of biochar continues to gain much
attention to improve the quality of agricultural soil in
an ecologically friendly pattern (Ayaz et al., 2021).
The application of biochar in agriculture is due to its
function to increase soil capacity to store water,
improve the structure, reduce bulk density, increase
pH, enhance electrical conductivity (EC), and cation
exchange capacity (Cybulak et al., 2021). Changes in
physical and chemical characteristics are
automatically followed by improvements in biological
characteristics and soil nutrients (Hossain et al., 2020).
To improve the performance of biochar in soil
quality, it is often combined with various inorganic
and organic liquid fertilizers, microbial inoculants, or
compost (Peng et al., 2021; Tian et al., 2021). A
previous study has discovered that biochar combined
with nutrient-rich organic matter through co-compost
increased the agronomic value by becoming richer in
NO3-, K, Ca, Si, Fe, and Al content, with higher C/N
values than the biochar matrix (Hagemann et al.,
2017). The application of biochar mixed with liquid
organic fertilizer from compost leachate containing
rich nutrients, namely N, P, K, Ca, and S, as well as
humic substances into the soil improved the
performance of soil microbial metabolic activity,
water, and nutrient retention, and plant growth
(Holatko et al., 2021).
The strategy of enriching biochar as a source of
biofertilizer and soil conditioner to achieve nutrient
and water of plant needs, efficient use of fertilizers,
and increase the population of beneficial soil biota to
improve crop productivity is still being developed
(Ndoung et al., 2021). This includes the use of liquid
extracts and sap of seaweed in agriculture as liquid
organic fertilizers that can be applied through the
leaves or irrigation (Hussain et al., 2021). It was
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discovered that the sap of Kappaphycus alvarezii (K-
sap) contains plant growth-promoting substances,
which include indole 3-acetic acid (IAA), gibberellin
(GA3), kinetin, and zeatin, as well as plant macro and
micronutrients, namely N, P, K, Na, Ca, Mg, S, Cu, Fe,
Mn, and Zn (Pramanick et al., 2020). Kappapychus
alvarezii is cultivated as the main commodity in the
coastal waters of Southeast Sulawesi (Aslan et al.,
2021). Its fresh biomass stores two products of
economic value, which include sap and carrageenan
(Mantri et al., 2017). In the current era of
intensification of agricultural ecology, the sap of K-sap
can be developed as a source of biofertilizer.
Therefore, this study aimed to (i) examine the
character of biochar enriched with K-sap from
different concentrations and (ii) analyze the soil
quality character after two weeks of treatment with the
K-sap enriched biochar
Materials and Methods
Soil collection and analysis
The soil samples of Ustults suborder (Soil Survey
Staff, 2014) used were taken at a depth of 10 cm from
upland rice fields of farmers in Wolasi, South Konawe
Regency, Southeast Sulawesi Province, at
122o29'33.14" east longitude and 4o11'29.27" south
latitude.
The samples were wind-dried, sieved at < 5 mm
per hole, and the soil that passed through was stored
until treated. Before the pot experiment, the soil
sample was analyzed at the Laboratory of the
Indonesian Soil Research Institute. The soil particle
fractions were determined using a pipette method. Soil
pH using a pH meter and electrical conductivity (EC)
using a conductivity meter were measured in 1:5 soil-
water suspensions. Organic C was estimated by the
Walkley and Black method. Total nitrogen was
assessed with the Kjeldahl method. A total of P2O5 and
K2O were estimated by 25% HCl extraction method.
Cation exchange capacity (CEC) was quantified by 1
N NH4OAc (pH 7) method, and exchangeable Al3+ and
H+ by 1 N KCl extraction method. Base saturation was
calculated as the sum of base cations divided by CEC
(Bélanger et al. 2006). The physicochemical
characteristics of the soil were 11% sand, 61% silt,
28% clay, 4.2 pH, 0.18 dS/m EC, 1.28% organic C,
0.11% total N, 12 C:N ratio, 32 mg/100 g total of P2O5,
7.0 mg/100 g total of K2O, 8.56 cmol/kg CEC, 15%
base saturation, 3.47 cmol/kg Al3+ and 0.53 cmol/kg
H+.
Site and experimental design
The experiment was carried out in the Greenhouse,
Faculty of Agriculture, Halu Oleo University, from
March to May 2021. The treatments consisted of a
biochar dose and a K-sap concentration factor. The
dose of biochar was based on the weight before it was
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enriched with K-sap to the weight of the soil, and is
expressed as % (biochar weight/soil weight).
The dosage levels of biochar were tested at 0%,
5%, and 7.5% (Kocsis et al. 2020), as well as 0%, 5%,
10%, and 15% K-sap concentrations (Singh et al.
2016). The combinations of these two factors were as
follows 0% biochar (B0), 5% biochar-enriched with
0% K-sap (B1), 5% biochar enriched with 5% K-sap
(B2), 5% biochar enriched with 10% K-sap (B3), 5%
biochar enriched with 15% (B4), 7.5% biochar
enriched with 0% K-sap (B5), 7.5% biochar enriched
with 5% K-sap (B6), 7.5% biochar enriched with 10%
K-sap (B7), and 7.5% biochar enriched with 15% K-
sap (B8). The nine treatments were arranged in a
randomized block design with three replicates.
Biochar production
Rice straw was burned imperfectly in a pyrolisator
made of drums at a 365 °C temperature until biochar
was formed. The biochar powder that passed through
the sieve of <150 µm/opening was stored in plastic
containers until enriched with K-sap.
Preparation of K-sap
Fresh seaweed, K-sap was obtained from the
cultivation area of local seaweed farmers in Bungin
Village, South Konawe Regency, Southeast Sulawesi
Province. The seaweed cleaned with seawater was cut
into 1-2 cm in size, mashed using a kitchen blender,
and the pulp was filtered with a cotton cloth (Shah et
al., 2013). The dripping sap was collected in a plastic
container and stored at 5 °C as stock until use. A total
of 25 mL of the sub-sample was filtered using
Whatman No. 42 for analysis by Inductively Coupled
Plasma-Optic Emission Spectroscopy (ICP-OES). The
K-sap stock contains 0.06 ppm Zn, 76.90 ppm Fe,
0.10% Mn, 0.67% Mg, 0.06% Ca, 0.10 ppm Cu, 0.85
% Na, 0.68% K, 0.02% P, and 0.40% S.
Enrichment of biochar with K-sap and analysis
K-sap stock was diluted using water to concentration
levels (v/v) of 0%, 5%, 10%, and 15%. Subsequently,
each concentration was mixed with biochar (500 mL
kg-1) until homogeneous. The labels for biochar
enriched with 0%, 5%, 10%, and 15% K-sap are B-
S0%, B-S5%, B-S10%, and B-S15%, respectively.
Each biochar enriched with K-sap was analyzed for
surface structure and chemical element composition
using SEM-EDS (Scanning Electron Microscopy-
Energy Dispersive Spectroscopy). The surface
functional groups were also evaluated with Perkin
Elmer Spectrum Two's FTIR-UATR (Fourier
Transform Infrared-Universal Attenuated Total
Reflectance), and crystalline phase using X-Ray
Diffraction (XRD).
Incorporation of K-sap enriched biochar to the soil
A total of 10 kg of soil that passed through a <5 mm
sieve/hole was mixed with K-sap enriched biochar
Open Access
until evenly distributed and was put into each
experimental pot, drained with tap water until
saturated, and incubated. During the incubation, soil
moisture was monitored using a moisture meter. When
the moisture content was below 35%, the soil was
moistened using tap water until it reached the desired
level.
Soil sampling and analysis
After incubation for two weeks, soil samples were
taken from each pot to analyze the chemical properties
of the soil, namely pH, EC, organic C, ammonium
(NH4+), nitrate (NO3-), available P, CEC, exchangeable
K, Ca, Mg, and Na. Soil biological characteristics
observed included the density of total bacteria, total
fungi, arbuscular mycorrhizal fungi (AMF) spores, and
flagellates. The total soil bacteria and fungi were
estimated using the MPN method. A 1 g of soil was
suspended in 9 mL of distilled water, and a series of
dilutions of the suspension were made from 10-1-10-7.
For total bacteria, 100 µL of the 10-6-10-7 series
suspension was evenly distributed on Tryptic Soy
Broth agar plates, and the plates were incubated in the
dark at room temperature for three days. Moreover, for
total fungi, 100 µL of suspension series 10-4-10-5 was
spread on Potato Dextrose Agar plates, incubated in
the dark under room temperature for seven days, and
the colony units formed for each plate were counted
(Vieira and Nahas, 2005; Pang et al., 2020). A total of
50 g of soil was suspended in 500 mL of water, filtered
in a stratified series downward from 2,000 µm, 200
µm, and 38 µm per opening. The spores retained on a
200 µm filter and 38 µm were transferred into tubes
containing 20% and 60% concentration gradient sugar
solution and centrifuged at 2,000 rpm for 3 minutes. At
the end of the spin, the solution containing the spores
was poured into a 38 µm top filter, and water was
added until all the sugar was rinsed off. The spores
were flowed using distilled water into a petri dish
equipped with a grid measuring 1 mm x 1 mm, counted
under a stereo dissecting microscope, and abundance
was estimated (Kilowasid et al., 2021).
The abundance of flagellates was estimated
through the direct counting method (Syaf et al., 2021).
Subsequently, one gram of fresh soil was suspended
with 5 mL of distilled water in a 5 cm diameter petri
dish. The suspension was left for 30 seconds until the
soil particles spread evenly and heavy sizes settled at
the bottom of the petri dish. A total of 15 µL of the
suspension was dropped into a hemocytometer, the
chamber was placed under a microscope at 400x
magnification, and visual cells swimming in 16
squares (0.0025 m2/squared) were counted.
Statistical analysis
All soil chemical and biological data were subjected to
ANOVA. When p<0.05, Duncan Multiple Range Test,
cluster analysis, principal component analysis (PCA),
and bivariate correlation analyses were applied.
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Results indicated an alternation with a higher concentration.

Surface morphology of K-sap enriched biochar
The surface structures of biochar-enriched with K-sap
at concentrations of 0% (B-S0), 5% (B-S5), 10% (B-
S10), and 15% (B-S15) were illustrated using SEM
image as shown in Figure 1. The surface morphology
of each biochar showed the presence of pores arranged
in a relatively similar pattern. The difference was the
inner side and surface of the pores of biochar enriched
with BS-5, BS-10, and BS-15 K-sap showed the
presence of dried sap droplets. The density of the dry
heap was more visible with the increasing
concentration of K-sap. EDS analysis of the
composition of C, O, P, K, Na, Mg, Ca, Si, Fe, Al, Cl,
and S in biochar enriched with K-sap from different
concentrations is illustrated in Table 1. Based on the
results, there was an increase in the C content, which
The O content in the enriched biochar increased at 5%
K-sap concentration (BS-5). Meanwhile, it was
discovered that a further increase in the SAP
concentration reduced the surface O and Al content
below the unenriched biochar.
The P content in B-S15 was higher, while the
concentrations of 5% and 10% were lower than that of
the unenriched. The content of K and S in B-S10
increased more in B-S15, while B-S5 was lower
compared to B-S0. The highest content of Na and Mg
was detected in B-S10, while at 5% K-sap
concentration, it was lower than the unenriched
biochar. In B-S15, the content of Mg was higher than
that of unenriched biochar. The content of Fe and Cl
also increased with a higher concentration of K-sap,
while Si in enriched biochar was lower compared to
the unenriched.

(A) (B) (C) (D)

Figure 1. SEM image of surface structure morphology of biochar enriched with K-sap from different
concentrations (A) B-S0; (B) B-S5; (C) B-S10; and (D) B-S15.

Table 1. Elemental composition (%, w w-1) of biochar enriched with K-sap from different concentrations.
Elements Biochar enriched with K-sap
B-S0 B-S5 B-S10 B-S15
C 12.90 23.46 17.32 23.58
O 43.17 43.66 36.77 38.99
P 1.37 0.98 1.02 1.41
K 2.22 2.01 5.00 3.90
Na 0.42 0.41 0.44 0.35
Mg 0.87 0.80 1.21 0.89
Ca 4.82 5.50 3.92 12.88
Si 32.07 20.82 30.92 14.52
Fe 0.65 0.78 1.28 1.62
Al 0.69 0.74 0.70 0.61
Cl 0.30 0.46 0.65 0.67
S 0.52 0.38 0.76 0.60
Notes: B-S0 is biochar enriched with K-sap 0% concentration; B-S5 is biochar enriched with K-sap 5% concentration; B-S10
is biochar-enriched with K-sap 10% concentration; B-S15 is biochar enriched with K-sap 15% concentration.

FTIR of biochar enriched with K-sap
FTIR spectroscopy detected that the adsorption band
range of B-S0 was from 454.76 to 3,383.78 cm-1,
which was distributed into 10 peaks. For B-S5 it was
450.98 to 3,354.79 cm-1, B-S10 was 453.73 to 3,369.02
cm-1, and B-S15 was 454.73 to 3,368.61 cm-1, each of
which was distributed into nine different peaks as
shown in Figure 2. The peak range from 3,384 to 3,354
cm-1 showed that biochar is associated with functional
groups, which represent stretching of the hydroxyl
group (-OH). The peak intensity from 1,579 to 1,587

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cm-1 represents the presence of aromatic structures in
all these biochar treatments. Furthermore, the peak
intensity of biochar (B-S0) which indicates the
functional group character for the band of carbonyl (C-
O) vibration, increased with the presence of K-sap.
The peak between 1,410 and 1,412 cm-1 indicates the
presence of functional groups of –C-C- and –C-H
vibration bands in biochar. The results showed that the
addition of K-sap decreased the peak intensity of the –
C-C- and –C-H vibration bands in the test biochar.
Peak spectral FTIR observed between 1,030-1,064
cm-1 shows the presence of a band of functional groups
Figure 2. Absorption band characteristic and peak wave number of biochar enriched with K-sap from different
concentrations.
The absorption characteristics ranged from weak to
moderate. The shift from B-S0 treatment at the peak of
794.69 and 778.60 cm-1 showed the double band and
weak, B-S5 treatment at the peak of 788.12 cm-1, B-
S10 treatment at 793.45 cm-1, and B-S15 at 792.04
cm-1 was a single and medium band. Absorption
characteristics for the weak band shifted from B-S0 at
the crest of wave 693.70 cm-1 was weak, while B-S5
treatment at peak 666.40 cm-1 was rather weak. For B-
S10, the wave peak shifted to a weak 712.13 cm-1, and
712.22 cm-1 for B-S15. Furthermore, B-S0 at the peak
of 527.80 cm-1 was moderate, B-S5 at 543.12 cm-1 was
weak, B-S10 at 543.00 cm-1 was rather weak, and B-
S15 at 543.00 cm-1 was very weak. Characteristics for
absorption sharp band of B-S0 at peak 454.76 cm-1, B-
S5 at peak 450.98 cm-1, B-S10 at peak 453.73 cm-1,
and B-S15 at peak 454.63 cm-1, respectively, were
strong.
XRD of biochar enriched with K-sap
The wave diffraction peaks reflecting the possible
crystalline material in each biochar-enriched K-sap
identified using XRD are illustrated in Figure 3. It was
discovered that the peak elevation pattern from the
Open Access
associated with C-O and P-O symmetric stretching
vibrations from phosphate residues. The intensity of
the absorbance band spectra increased with the
presence of K-sap in the biochar. The weak band with
a peak intensity of 913.46 cm-1 was only observed at
B-S0, while the spectra at B-S5, BS-10, and B-S15
were not detected. From 876 to 873 cm-1, the weak
band intensity decreased with the addition of K-sap
into biochar which was observed in all treatments.
Weak double bands at the intensity of 794.69 and
778.60 cm-1 were observed in B-S0, and enrichment of
biochar with K-sap eliminated one peak band.
four biochar enriched with different concentrations,
0%, 5%, 10%, and 15% of the K-sap was relatively
similar to each other. Peak elevation between 2-Theta
is 20.81° to 50.1°. The high peak that occurred at the
elevation of 2-Theta was from 26.64° to 29.38°, while
the other elevation peaks were small.
Effect on soil chemical characteristics
Biochar enriched with K-sap at different
concentrations significantly affected pH, EC, organic
C, total N, C:N ratio, available P, exchangeable K, Ca,
Mg and ammonium (NH4+) of the soil after 2 weeks of
treatment, except for nitrate (NO3-) and mineral
nitrogen (NH4++NO3-), and exchangeable Na. From
Table 4, it was also discovered that pH, EC, C:N ratio,
exchangeable K, and Si in B7 were the highest, while
the lowest content occurred in B0. Soil pH from high
to low according to treatment sequentially were B7,
B8, B6, B5, B1, B3, B4, B2, and B0. The difference in
soil pH at B7 compared to B0, B1, B2, B3, B4, and B5
was significant, except for B6 and B8. The difference
in pH in treatment B8 compared to B2 and B0 was
significant, while that of B6, B5, B4, B3, and B1 was
insignificant.
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Figure 3. X-ray diffraction pattern of K-sap enriched biochar of different concentrations.
The variation in pH at B5 compared to B2 and B0 was
significant, except for B4, B3, and B1. There was also
a significant difference in soil pH in treatment B2
compared to B0. Soil EC values from high to low were
based on sequential treatments, as stated in Table 3,
namely B7>B5>B6>B8>B2>B1> B4>B3>B0. The
difference in soil EC at B7 compared to B0, B1, B2,
B3, B4, B6, and B8 was significant, but B5 was
insignificant. In B5, there was a significant difference
in EC compared to B0. Furthermore, B6 and B8
compared to B0 were significantly different, while the
other treatments were not, except for B7. The order of
organic C from high to low was
B6>B7>B5>B8>B3>B1>B2>B0, and the total N was:
B6>B8=B7=B5>B4=B3=B2=B1>B0 (Table 3). The
variation between organic C and total soil N in B6
compared to other treatments was significant, except
for B5, B7, and B8. Organic C and total soil N in both
B3, B4, B1, and B2 compared to B0 were not
significantly different. The order of levels of the C:N
ratio from high to low was as follows
B7=B5>B8=B6>B4=B3=B2=B1>B0. There was also
a significant difference in C:N ratio in B7 compared to
B0, B1, B2, B3, and B4. The level of soil NH4+ content
according to the treatment in the sequence was
B0>B5>B7>B8=B6>B3>B2=B4>B1. The difference
between NH4+ in B0 compared to other treatments was
significant. The order for available P was as follows:
B6>B8>B2>B7>B4>B5>B3>B1>B0. The variation
in available soil P in B6 compared to B0, B1, B3, B4,
and B5 was significant, while the other treatments
were insignificant. Available P at B0 was more
significant than other treatments.
The order of exchangeable Ca levels was
B6>B8>B1>B3>B5>B4>B7>B2>B0. The difference
in exchangeable Ca in B6 compared to B0, B2, B3, B4,
B5, and B7 was significant, except for other
treatments. Exchangeable Mg sequentially as follows:
B6>B7>B5>B8>B1>B3>B4>B2>B0. The
exchangeable Mg in B6 compared to B0 and B2 was
Open Access
also different. The exchangeable K, according to the
treatment sequentially, was as follows:
B7>B5>B6>B8>B2>B4>B1>B3>B0. K
exchangeable in B7 compared to other treatments was
significantly different, except for treatment B5. The
content of exchangeable K in B0 compared to B5, B6,
and B8 was significantly different, except for B1, B2,
B3, and B4. Furthermore, the sequence of
exchangeable Na content between treatments
sequentially was as follows:
B2>B7>B6>B5>B4>B1>B8>B3>B0. The content in
B2 compared to B0, B3, and B8 was different, while
there was no variation between B1, B3, B4, B5, B6,
B7, and B8. Si content according to the treatment
sequentially was as follows:
B7>B6>B8>B5>B1>B3>B4>B2>B0. The difference
in Si content in B7 compared to B0, B1, B2, B3, and
B4 was significant. Si content in B0 compared to B2,
B3, and B4 was not significantly different, while
compared to B1, B5, B6, and B8 were significantly
different. The variation between treatments B1, B3,
B4, B5, B6, and B8 was also not significant.
Effect on soil biological characteristics
The effect of biochar enriched with K-sap at different
concentrations on AMF spore and flagellate
abundance was significant. The order of abundance of
AMF spores according to treatment was
B0>B2>B3>B1>B7>B5>B6>B4>B8, as shown in
Figure 4. A significant difference was discovered in
total AMF spores in B0 compared to other treatments.
Total AMF spores in B2 compared to B8 also varied.
However, there was no difference in total AMF spores
between treatments B3, B1, B7, B5, B6, B4, and B8.
The order of flagellate abundance levels between
treatments was as follows:
B3>B2>B6>B1>B4>B8>B7>B5>B0. In Figure 4B,
the variation in the abundance of flagellates in B3
compared to B0 was significant, while others were
insignificant.
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Table 2. Soil chemical characteristics after two weeks incubated with K-sap enriched biochar.
Parameter Treatments
B0 B1 B2 B3 B4 B5 B6 B7 B8
pH(H2O. 1:5) 4.77 a 6.27 bc 5.90 b 6.17 bc 6.10 bc 6.50 c 6.67 cd 7.20 d 6.67 cd
EC (dS/m) 0.13 a 0.40 ab 0.40 ab 0.34 ab 0.36 ab 0.72 bc 0.56 b 1.07 c 0.53 b
Organic C (%) 1.31 a 2.19 b 2.14 b 2.22 b 2.19 b 2.94 c 3.01 c 2.97 c 2.79 c
Total N (%) 0.12 a 0.18 b 0.18 b 0.18 b 0.18 b 0.22 c 0.23 c 0.22 c 0.22 c
C:N ratio 11.00 a 12.00 ab 12.00 ab 12.00 ab 12.00 ab 13.33 c 13.00 bc 13.33 c 13.00 bc
Available P (ppm) (x102) 0.16 a 0.92 b 1.10 bcd 0.93 b 0.98 bc 0.95 b 1.25 d 1.05 bcd 1.21 cd
Exchangeable Ca (cmol/kg) 0.82 a 6.56 bcd 4.33 b 6.11 bc 5.31 bc 5.38 bc 9.34 d 4.91 b 8.27 bcd
Exchangeable Mg (cmol/kg) 0.26 a 1.22 bc 0.84 b 1.06 bc 0.89 bc 1.35 bc 1.49 c 1.40 bc 1.27 bc
Exchangeable K (cmol/kg) 0.11 a 1.18 ab 1.62 abc 1.14 ab 1.38 abc 4.04 cd 2.01 bc 7.53 d 1.94 bc
Exchangeable Na (cmol/kg) 0.25 a 0.32 a 0.56 a 0.26 a 0.33 a 0.37 a 0.42 a 0.50 a 0.27 a
NH4+ (mg/kg) 59.00 b 5.67 a 6.33 a 6.67 a 6.33 a 9.33 a 7.33 a 8.00 a 7.33 a
NO3- (mg kg-1) (x102) 2.84 a 4.42 a 6.54 a 5.72 a 6.63 a 12.22 a 9.03 a 9.61 a 9.63 a
Mineral N (mg/kg) (x102) 3.43 a 4.48 a 6.60 a 5.79 a 6.69 a 12.31 a 9.10 a 9.69 a 9.71 a
Si (mg/kg) (x102) 1.17 a 3.29 b 2.58 ab 3.03 ab 2.81 ab 4.21 bc 4.59 bc 5.34 c 4.53 bc
Notes: Numbers with different letter superscripts in the same row indicate significantly different indications according to Duncan Multiple Range test (p < 0.05).

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The difference in the number of flagellates between between the treatment of K-sap enriched biochar on
treatments B2, B6, B1, B4, B8, B7, and B5 was not total bacteria is shown in Figure 4C, and total fungi are
significant, except for B0. The insignificant difference shown in Figure 4D.

Figure 4. A) Total AMF spores; B) Total flagellates; C) Total bacteria; D) Total fungi
Notes: Different letters above the bar indicate significant differences according to Duncan Multiple Range test
(p<0.05).

The similarity of soil quality among treatments
Out of the 14 chemical and 4 biological parameters of
soil quality tested using ANOVA, there were 12
chemical and 2 biological characteristics that showed
significant effects. Cluster analysis using the paired
group algorithm method with the Bray-Curtis index on
the affected parameters and obtained four soil quality
clusters (Figure 5A), namely cluster I formed by B0,
cluster II by B1, B2, B3, and B4, cluster III by B6, and
cluster IV by B5, B7, and B8. Moreover, cluster I soil
quality had a similarity index of Bray-Curtis <0.60
with soil quality from others. Cluster III (B6) had a
similarity index of Bray-Curtis <0.80 with soil quality
from other clusters. Cluster II soil quality (B1, B2, B3,
and B4) had a high Bray-Curtis similarity index
ranging from 0.89 to 0.97. The Bray-Curtis index of
similarity in soil quality between treatments in cluster
IV (B7 and B8) was 0.91. The Bray-Curtis similarity
index between soil quality from B7 to B1 and B8 was
0.80 and 0.90, respectively. The similarity of soil
quality between B8 and B1, B3, B4, and B5 were 0.85,
0.82, and 0.94, respectively.
Principal component analysis (PCA) and bivariate
correlation analysis
The relationship between the eigenvalues and the
principal component (PC) of soil quality parameters
that showed a significant effect analyzed by PCA is
illustrated in Figure 5A. The result showed that there
was a correspondence between decreasing eigenvalues
and increasing PCs (Figure 5B). The increase in PCs
up to 4 significantly reduced the eigenvalues to PC8.
Out of the 8 PCs built, only 3 had eigenvalue >1 and
variance >5%, namely PC1, PC2, and PC3, one PC,
namely PC4, with eigenvalue <1 and variance >5%,
and 4 other PCs had eigenvalue <1 and variance <5%,
as shown in Table 3. It was also discovered that PC1 -
PC4 showed 97.957% of the total variance of the total
selected soil quality attribute data tested. PC1
explained 65.145% of the soil quality variance, which
indicates that the eigenvalue was 846.883.
Furthermore, PC2 explained 20.155% of the soil
quality variance and suggested that the eigenvalue was
262,013. PC3 described 11.503% of the variance, and
the indicated eigenvalue was 149,534. PC4 explained

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14.443% of the variance, with an eigenvalue of 0.188.
Loading factors of pH, EC, total N, C:N ratio, available
P, exchangeable Mg, and Si were positively correlated,
while ammonium and total spore AMF were
negatively correlated with PC1 as presented in Table
3. The organic C, Ca and exchangeable K were
positively correlated with PC2. The number of
flagellates negatively correlated with PC3.
Subsequently, according to Table 4, the
interrelationship between parameters (loading factor)
of soil quality correlated with PC1. From the
Spearman rho correlation analysis (Table 4) showed
that soil pH significantly positively correlated with
EC, total N, C:N ratio, exchangeable Mg, and Si.
However, it negatively correlated with total spore
AMF, while available P and ammonium were not
significant. There were significantly positive
correlations between available P with total N, C:N
ratio, exchangeable Mg, and Si, with total AMF spores
being negatively significant, as indicated in Table 4.
Among parameters (factor loading) of soil quality in
PC2, Spearman rho correlation analysis showed that
organic C positively correlated with exchangeable Ca
and K.
The relationship between the number of
flagellates (factor loading PC3) with available P
(factor loading PC1), exchangeable Ca (factor loading
PC2) and exchangeable Mg (factor loading PC1) was
positively significant and negatively correlated with
ammonium ions (factor loading PC1) and total AMF
spores. Factor loading of PC4, exchangeable Ca with
EC as well as exchangeable Mg and K (factor loading
PC2) were positively correlated as presented in Table
4.

(A) (B)
Figure 5. (A) Dendrogram of soil quality among different treatments; (B) Scree plot of soil quality from different
treatments.

Table 3. Summary of Principal Component Analysis using the correlation matrix

PC1 PC2 PC3 PC4 PC5 PC6 PC7 PC8
Eigenvalue 846.883 262.013 149.534 0.188 0.110 0.065 0.036 0.017
% variance 65.145 20.155 11.503 1.443 0.849 0.499 0.277 0.128
Loading Factors:
pH 0.327 -0.138 0.116 0.161 -0.244 -0.389 0.207 -0.253
EC 0.255 -0.175 0.437 0.629 0.213 -0.090 -0.366 0.072
Organic C 0.160 0.547 -0.001 0.007 -0.013 -0.051 -0.075 -0.003
Total N 0.335 -0.019 0.120 -0.301 0.243 0.262 0.025 -0.064
C:N ratio 0.309 -0.085 0.304 -0.241 0.292 0.476 -0.146 -0.173
Available P 0.317 -0.044 -0.253 -0.148 0.551 -0.297 0.304 0.459
Exchangeable Ca 0.160 0.546 -0.010 -0.007 -0.029 -0.062 -0.049 0.004
Exchangeable Mg. 0.332 -0.028 0.068 -0.068 -0.616 0.378 0.088 0.456
Exchangeable K. 0.176 0.529 0.046 0.100 0.014 -0.075 -0.133 -0.036
NH4+ -0.279 0.180 0.401 -0.103 0.110 0.081 0.433 -0.365
Si 0.319 -0.070 0.273 -0.017 -0.156 -0.219 0.503 -0.098
AMF -0.307 0.160 0.244 0.392 0.155 0.285 0.400 0.445
Flag. 0.232 -0.001 -0.568 0.478 0.072 0.408 0.272 -0.369

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Table 4. Spearman rho correlation between parameters of soil quality after two weeks incubated with biochar enriched with K-sap.
pH EC Organic C Total N C:N ratio Available Exchangeable Exchangeable Exchangeable NH4+ Si AMF
P Ca Mg K
EC 0.55
Organic C 0.71 0.73
Total N 0.70 0.70 0.97
C:N ratio 0.60 0.74 0.93 0.93
Available P 0.31 0.31 0.55 0.64 0.53
Exchangeable Ca 0.48 0.33 0.54 0.56 0.43 0.67
Exchangeable Mg 0.56 0.76 0.63 0.59 0.55 0.43 0.67
Exchangeable K 0.66 0.84 0.78 0.79 0.76 0.55 0.35 0.66
NH4+ -0.10 0.18 0.14 0.12 0.21 -0.28 -0.26 -0.12 0.06
Si 0.83 0.54 0.69 0.70 0.56 0.50 0.71 0.69 0.64 -0.13
AMF -0.45 -0.32 -0.46 -0.49 -0.45 -0.62 -0.56 -0.49 -0.49 0.26 -0.57
Flag. 0.07 0.04 0.13 0.12 0.00 0.52 0.52 0.44 0.14 -0.67 0.26 -0.35
Notes: The numbers in bold indicate a real correlation between the respective parameters. Numbers in bold without a sign indicate a positive correlation, and a minus sign (-) in front of a number
in bold indicates a negative correlation.

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Discussion
The characteristics of biochar enriched with K-sap
The SEM image in Figure 1 shows the inner side of
several pore spots of K-sap enriched biochar with
visible droplets of dried K-sap covering the surface of
the spots, while the unenriched was not visible. The K-
sap hatchlings contain polysaccharides with several
functional groups (Bhuyar et al., 2021; Perera et al.,
2021). The interaction between the elements of this
group and the chemical elements of the biochar can
cause a change in the functional group of the biochar
surface. FTIR showed that the functional groups
associated with OH-stretching, C-O and P-O
symmetric stretching vibration, C-O bending
vibration, C-H deformation, and absorption band of Si-
O of biochar changed after enrichment, as presented in
Figure 2. This indicated that the enrichment of biochar
with K-sap changes the band adsorption characteristics
of biochar (Liu et al., 2019). The presence of hydroxyl
groups (-OH) in cellulose, hemicellulose, or
polysaccharides alters the ability of biochar and dry K-
sap to absorb water (Baskararaj et al., 2019; Adeniyi et
al., 2020). The absorption band strength of the
functional groups associated with the OH-stretching of
the enriched biochar was wider than that of the
unenriched, in which the strength increased with the
concentration of the K-sap (Figure 2). This indicated
that the ability of surface biochar to absorb water is
reduced to the concentration of sap.
The decrease in the stretching vibration of the
hydroxyl bond (-OH) is related to the gel-like droplets
of the sap drying on the enveloped area and forming a
film that has physical and mechanical properties
similar to plastic (Sudhakar et al., 2021). This dry gel-
like layer functions as a barrier to prevent water from
being absorbed by the surface of the biochar, thereby
inhibiting the absorption of biochar into water vapor in
free air (Hegde et al., 2021). The presence of K-sap can
change the absorption band of biochar at a wave
number of 1,030.44 cm-1 with a strong character and
reduced sharpness. The C-O and P-O symmetric
stretching vibration functional groups of unenriched
biochar interact with the C-O of the polysaccharide
groups in the K-sap. This leads to the disappearance of
the P-O symmetric stretching vibration bands
generated from the phosphate residues from the
pyrolysis of the polysaccharides.
The bending C-O vibration band at wave number
913.46 cm-1 of biochar also disappeared in the
presence of sap due to the strong interaction between
the C-O groups of the sap and biochar. The double
band C-H deformation of biochar disappeared, and
wave absorption increased when enriched with sap.
The absorption band of the Si-O group of biochar in
the wave number ranging from 543 to 712.22 cm-1 was
weakened in the presence of the K-sap, as shown in
Figure 2. The alternating wave numbers from 1,030.44
to 1,063.45 cm-1, 788.12 to 794.69 cm-1, and 666.40 to
Open Access
712.22 cm-1 indicated that the K-sap concentration
affected the energy absorption behavior of biochar,
with the highest value occurring at K-sap 10%
concentration, except for peak 913.46 cm-1. Moreover,
an increase or decrease in the absorption spectrum
strength, the formation of a new spectrum, and
functional groups can also occur for products with
feedstock derived from a mixture of seaweed and
husks (Wang et al., 2016).
In this study, the enrichment of biochar with K-
sap did not change the crystalline structure of the
parent materials. The peak that occurred at an elevation
of 2-Theta 26.64° to 29.38° in Figure 3 indicated the
presence of SiO2 and CaCO3 components (Zhang et al.,
2018). This was confirmed by the SEM-EDS results
(Table 1), which detected more dominant Si and Ca
elements than other metal-forming crystal elements
such as Al. Fe. K. Mg. and Na. Compositional
variations in nutrient content and functional groups of
each enriched biochar provided an indirect description
of the function in changing soil fertility controllers,
including pH, and exchange capacity, as well as
retention of cation when applied to the soil (Chen et
al., 2019).
Effect of biochar enriched with K-sap on soil quality
After two weeks of soil incubation, four soil quality
groups were created. These include the group without
biochar, the group with a dose of 5% biochar, 7.5%
biochar enriched with 5% K-sap and the 7.5% enriched
with K-sap of 10% and 15% (Figure 5A). PCA results
showed that pH, EC, organic C, exchangeable of Ca,
K, and Mg, as well as ammonium, Si, AMF, and
flagellates, correlated with biochar treatment. The soil
that was treated with B7 had increased pH and EC of
1.51 and 8.23 times higher than without biochar. The
pH value that was treated with B7 was significantly
higher than that of the soil with only biochar. This
indicated that the presence of K-sap contributed to an
increase in the OH-ion content in the soil solution. This
is closely related to the FTIR results, which detected
that biochar enriched with 10% K-sap concentration
had a strong intensity, with a wave number 3.369.02
cm-1 higher than that of unenriched or enriched biochar
with 5% and 15% K-sap at the peak area for OH
(Figure 2).
The presence of CaCO3 crystals in the biochar as
detected using XRD in Figure 3 played a role in
increasing the pH of the acid soil. The EC value of soil
treated with B7 was also higher than that of unenriched
biochar, as shown in Table 2, which had a relationship
with the high pH value. The Spearman rho correlation
showed a significant positive relationship between the
pH and EC of soil. The highest soil EC value
(1.07±0.46 dS/m) achieved from all treatments was
still below the threshold for stress conditions which
were very suitable for the growth and yield of all crops
(Shahid et al., 2018). The exchangeable K and Si in the
B7 treatment were the highest compared to others,
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except for the B5 treatment for K, and B5, B6, and B8
for Si. Furthermore, the exchangeable K that occurred
in treatment B7 was consistent with the K content in
biochar enriched with K-sap at a concentration of 10%
compared to other treatments, which were the highest
(Table 1). The Si content in biochar enriched with K-
sap at 10% was higher than the concentration of 5%
and 15% as shown in Table 1, but there was no
significant effect of Si in soil between concentrations
of K-sap in the same dose of biochar. The peak
intensity detected in the range of 527.40 to 543.12
cm-1 and 666.40 to 712.22 cm-1 explained the Si-O
functional groups in biochar, both without and
enriched with K-sap (Figure 2). This indicated that the
changes in Si content in the soil are more promoted by
the amount of biochar added.
The results showed that biochar enriched with K-
sap significantly increased the organic C, total N, and
C:N ratio compared to the soil sample without biochar
(Table 2). Furthermore, the content of organic C and
total N in the soil treated with a dose of 7.5% biochar
without or with K-sap for all concentrations was higher
than that of biochar at a dose of 5%. The C:N ratio of
soil at a dose of 7.5% biochar without K-sap (B5), and
enriched with K-sap in 10% concentration (B7) was
1.21 higher than B0. The high soil C:N ratio that
occurred at a dose of 7.5% biochar enriched with K-
sap indicated that the treatment was able to increase
the organic C content of the recalcitrant. This
contributed to organic C stabilization and increased
soil C:N ratio. According to Holatko et al. (2021), the
application of biochar enriched with liquid organic
fertilizer can increase the total organic C, leading to C
stabilization and increasing the soil C:N ratio.
Meanwhile, there was no significant difference in the
content of organic C, total N, and C:N ratio of the soil
between treatments in the dosage of biochar without
and with enriched K-sap. This indicated that the
volume of K-sap from each concentration mixed with
biochar did not affect the increase in the content of
organic C, total N, and C:N soil ratio (El-Naggar et al.,
2019; Winarso et al., 2020).
Biochar increases the available P, exchangeable
K, Ca, and Mg of acid soils (Wu et al., 2020). In Table
2, the available P at the dose of 7.5% biochar without
or enriched was higher than other treatments, except
for 5% biochar enriched with 5% K-sap. The content
of available P and exchangeable Ca that occurred at the
dose of 7.5% enriched with 5% K-sap was the highest,
compared to the dose of 7.5% biochar without
enrichment, while the 7.5% enriched with 10% and
15% was not significant. These results showed that the
exchangeable Mg could be increased by the
application of biochar with and without K-sap.
Contents of organic C, total N, and C:N ratio were
often used as measures to describe the capacity of the
soil to provide energy for soil biota in the nutrient
recycling process in acid soils (Li et al., 2016).
Furthermore, it was discovered that there are positive
Open Access
correlations between organic C, total N, and C:N ratio
to available P, exchangeable Ca, Mg, K, and Si in the
soils, as shown in Table 4.
Table 4 shows that total C and N, as well as C:N
ratios, negatively correlated with total AMF spore due
to an increase in available P (Sivakumar, 2013; Lin et
al., 2020). Yu et al. (2021) discovered a significant
negative relationship between the total AMF spores to
the K content in the soil. Furthermore, the negative
correlation with K, Ca, Mg, and Si was in line with the
relationship between the total AMF spore with soil pH.
These results indicated that AMF spores are more
suitable for acid soils (Han et al., 2019). The release of
Si increased with soil pH up to 6.5, and concentrations
of Ca, Mg, Na, and K (Ngoc Nguyen et al., 2014). In
this study, the Si content of the soil with the addition
of 7.5% biochar enriched with 5% K-sap increased
more than the samples without treatment and those
with the 5% biochar as well as the enriched, as shown
in Table 2.
The Spearman rho correlation in Table 4
indicated a positive relationship between Si and pH in
soil (r = 0.83), Ca (r = 0.71), Mg (r = 0.69), and K (r =
0.64). It was discovered that increasing Si in the soil
can reduce P sorption, thereby improving the
proportion of available P increases in acid soil (Greger
et al., 2018). The Spearman rho correlation also
implies that there was a positive relationship (r = 0.50)
between the Si and available P in the soil. This
indicated that the availability of P in the soil can be
increased through the addition of biochar which can
improve the Si content in acid soils. Table 4 revealed
that there is a negative correlation (r = - 0.57) between
Si and the total AMF spore. This indicated that the
reduction in AMF spore abundance is associated with
an increase in pH and available P based on the higher
values of Si content in the acid soils.
Conclusion
The functional groups associated with the OH-
stretching of biochar enriched with K-sap were
stronger and wider than those without treatment. The
strength and width of the band increased with the
concentration of the K-sap. Biochar enriched with K-
sap improved the quality of acid soil within two weeks
of incubation. The doses of 7.5% biochar enriched
with K-sap 5% and 10% concentrations significantly
increased pH, exchangeable K, and Si, organic C, total
N, C:N ratio, available P, and exchangeable Ca and Mg
in acid soil. Furthermore, the abundance of flagellates
increased, while the total AMF spores decreased with
the addition of biochar with or without K-sap.
Acknowledgements
The authors thank the Directorate of Resources, Directorate
General of Higher Education, Ministry of Education,
Culture, Research and Technology for funding this study,
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contract number: 270/E4.1/AK.04.PT/2021. The authors S.M. and Ok, Y.S. 2019. Biochar application to low
also thank the National Research and Innovation Agency fertility soils: A review of current status, and future
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