Journal of Soil Science and Plant Nutrition

https://doi.org/10.1007/s42729-021-00583-0

ORIGINAL PAPER

Evaluation of a Biochar‑Based Slow‑Release P Fertilizer to Improve

Spinacia oleracea P Use, Yield, and Nutritional Quality
Cindy Sepúlveda‑Cadavid1 · Jhon H. Romero1 · Maicol Torres1 · Evelyn Becerra‑Agudelo1 ·
Julián E. López1,2

Received: 11 April 2021 / Accepted: 12 August 2021

© Sociedad Chilena de la Ciencia del Suelo 2021

Abstract
The aim of this study was to assess the effects of a biochar-based slow-release P fertilizer (BBSRPF) produced from a corn
stover-derived biochar (CSB), compared to mineral fertilization and co-application of CSB itself and mineral fertilization
on bioavailability of soil P and spinach production, as well as, foliar macronutrients (N, P and K), protein, and ascorbic acid
in plants.
First, CSB was produced from corn stover. Then, P loading process was done with a ­KH2PO4 solution for BBSRPF produc-
tion. Afterwards, a greenhouse experiment was performed for 70 days using a factorial design with the following treatments:
mineral NPK fertilization (F), CSB and mineral NPK fertilization (BC) and BBSRPF and mineral NK fertilization (P-BC).
After this, soil samples were collected and the plants were harvested to determine changes in soil physicochemical proper-
ties, bioavailability of soil P, leaf content of P, N, K, ascorbic acid, and protein. Besides, fresh and dry biomass, and crop
yield were also determined.
The P-BC treatment significantly increased bioavailable soil P by ~ 72% compared to F and BC treatments. Furthermore, P,
N, K, protein, ascorbic acid content, and yield of spinach pants were significantly increased by ~ 29, 52, 33, 20, 21, and ~ 25%
when P-BC treatment was applied, respectively.
Incorporation of P into the biochar matrix is an effective strategy to increase spinach P use efficiency in tropical soils. There-
fore, BBSRPF could totally replace conventional mineral P fertilizers without compromising crop yield, and increasing the
nutritional quality of spinach plants.

Keywords Spinach · Protein · Ascorbic acid · Mineral nutrition · Soil quality

1 Introduction compounds (Carvalho et al. 2019; Salehi et al. 2019). Due

Spinach (Spinacia oleracea) is a leafy vegetable which
belongs to the family Amaranthaceae. Which is well
known since its composition includes high nutritional value
Cindy Sepúlveda-Cadavid and Jhon H. Romero contributed
equally to this work.
* Julián E. López
jelopez@udem.edu.co; julian.lopez@colmayor.edu.co
1
Semillero de Investigación en Ciencias Ambientales—
SICA, Facultad de Arquitectura E Ingeniería, Institución
Universitaria Colegio Mayor de Antioquia, Carrera 78 # 65
– 46, 050034 Medellín, Colombia
2
Facultad de Ingenierías, Programa de Ingeniería
Ambiental, Universidad de Medellín, Carrera 87 N° 30‑65,
050026 Medellín, Colombia
to this, it has been reported that in USA spinach production
and consumption increased by 418% and 31%, respectively,
from 1970 to 2012 (Roberts and Moreau 2016). Moreover,
in Colombia, the production of leafy vegetables including
spinach has been of great importance in recent years, with
10,000 ha of land planted of this kind of crops, approxi-
mately. Producing about 18.1 ton ha−1 of spinach in 2017
(Casierra-Posada et al. 2021). Which may be associated to
the growing awareness of healthier lifestyles based on plant-
based diets and alertness of chronic diseases associated to
unhealthy nutritional habits, e.g., cardiovascular problems,
overweight, cancer to name but a few (Fan et al. 2014; Lara
et al. 2021).
On that basis, spinach plants are composed by several
substances, some of which have been reported to have ben-
eficial effects on human health. For instance, ascorbic acid is

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known as an important antioxidant agent present in spinach
plants, that can have a positive influence on the reduction of
risk of some diseases such as cancer, arteriosclerosis, cata-
racts and cardiovascular diseases (Bergquist et al. 2006).
Moreover, the increasing interest on vegetable proteins has
led to looking for the improvement of protein content in
vegetables, due to negative impacts of production of animal
proteins on the environment such as acidification, erosion,
eutrophication, biodiversity loss and climate change, related
to the use of land, water, fuels, fertilizers, pesticides and
animal waste production (Aiking 2011; Sabaté et al. 2015).
In addition to this, negative impacts on human health have
also been associated to high consumption of animal protein
sources, like obesity, hypercholesterolemia and higher prob-
ability of suffering from cancer (Lander et al. 2016; Marti-
Quijal et al. 2019).
Considering that environmental conditions and agronomi-
cal practices can influence the composition and the devel-
opment of spinach plants, it is necessary the researching
on agricultural products that lead to the improvement of
the crop nutritional value, as a result of the enhancement
of environmental factors associated to soil parameters and
nutrients availability (Salehi et al. 2019). In the same way,
these products should promote soil health, as safe food pro-
duction depends on it (Gallego and Olivero-Verbel, 2021).
Specifically, in acidic soils, the applied agronomical prac-
tices must take into account their limitations regarding crop
production potential (Bolan et al. 2003). For instance, for
the case of P behavior, this nutrient can be fixed by Fe and
Al oxides which restricts its availability. This reduces the
efficiency of P fertilization practices traditionally used (Zhu
et al. 2018). Furthermore, traditional practices do not con-
tribute to soil health improvement. On the contrary, these
can cause negative impacts on soil properties and environ-
mental dynamics (Bouwman et al. 2013; Lin et al. 2019).
Regarding this, several alternatives to traditional chemi-
cal fertilizers have arisen in recent years. For instance, bio-
chars are a target of an increasing interest as an alterna-
tive, since their high carbon content and their active surface
area have shown to have several positive impacts on soil
physicochemical and biological properties (e.g., pH, soil
enzymatic activities), crop yield and nutrition (e. g., plat
productivity, relative water content, content of N, P, and K,
as well as biochemical compounds), and the reduction of
toxic effects (e.g., oxidative stress) on plants due to heavy
metals in soils (Naeem et al. 2021; Turan 2019; Zubair et al.
2021). However, with respect to P bioavailability, the co-
application of biochar itself and phosphate fertilizers can be
counterproductive in some cases. For example, the applica-
tion of biochar can increase the capability of P fixation in
acidic soils, depending on application rates of biochar and
phosphate fertilization. Thus, P availability is reduced, com-
pared to neutral and alkaline soils, in which co-application
13
Journal of Soil Science and Plant Nutrition
of biochar and fertilizer improve the availability of this ele-
ment (Xu et al. 2014). In line with this, Chintala et al. (2014)
also found a similar behavior regarding P constrain in acidic
soils after biochar application. In contrast, Li et al. (2020)
and Eduah et al. (2019) have reported a decrease in the effi-
ciency of phosphate fertilization with the co-application
of biochar and traditional chemical fertilizers in neutral to
alkaline soils. These results suggest that there is still lack of
understanding on the influence of this fertilization alterna-
tive on P bioavailability management in crops. In the case of
spinach, studies such as Li et al. (2016) report a decrease in
P nutrition with the application of biochar in soil compared
to the use of chemical fertilizer. In addition, plant-derived
biochar generally has a limited concentration of bioavailable
P, making it uncompetitive for the supply of this nutrient
compared to other traditionally used sources (Li et al. 2020).
To overcome the limitations that biochars itself have
regarding the nutritional management of P in crops, the use
of P-laden biochar as slow-release fertilizers has been sug-
gested as an alternative (Volf and Rosolem 2020). In tropi-
cal soils it has been found that biochar-based slow-release
P fertilizers (BBSRPFs) have shown a superior behavior
to improve P bioavailability in soils and crop production,
compared to conventional mineral fertilizers (e.g., triple
superphosphate) (Carneiro et al. 2021; Lustosa Filho et al.
2020; Pogorzelski et al. 2020). Nevertheless, to the authors
knowledge, the effect of BBSRPF to supply 100% of the
nutritional demand for this element in important leafy crops
such as spinach and its impact on production and plant nutri-
tional quality has not been widely explored. Hence, further
research is needed in order to evaluate the effectiveness of
BBSRPFs compared to mineral fertilizers and co-application
of biochar and mineral fertilizer regarding spinach yield,
nutritional content (N, P, and K) and biochemical compound
of interest (protein, ascorbic acid). This, aiming for more
sustainable agronomic practices equated with conventional
fertilizers schemes in terms of crop production. Specially in
Colombian agricultural soils, in which insufficient fertiliza-
tion and progressive depletion of key nutrients are limiting
factors for production of vegetable crops. For instance, the
extraction rates of P exceed soil nutrient availability by 20%,
approximately (Bojacá et al. 2010).
Accordingly, this study evaluated the effects of a BBSRPF
produced from a corn stover derived biochar (CSB), on
P use, yield and nutritional quality of spinach plants in
comparison with traditional mineral NPK fertilization and
co-application of mineral NPK fertilizer and corn stover
derives biochar itself. We hypothesize that the application
of BBSRPF on acidic tropical soil improves crop produc-
tion and its nutritional quality, through the enhancement of
P bioavailability and other soil physicochemical properties.
The aims of this study were (i) to assess BBSRPF capacity
for improving soil properties and the bioavailability of P,
Journal of Soil Science and Plant Nutrition
(ii) to compare the effect of the application of BBSRPF, a
traditional mineral NPK fertilizer, and the co-application
of CSB and traditional mineral NPK fertilizer on spinach
nutritional quality (N, P, K, protein and ascorbic acid) at
leaf level, and (iii) determine the effect of these treatments
on spinach yield. In order to support our hypothesis, a green-
house experiment was conducted comparing the application
of mineral NPK fertilizer, CSB with mineral NPK fertilizer,
and BBSRPF with mineral NK fertilizer on spinach leaves
content of N, P, K, protein and ascorbic acid, and crop yield.
Additionally, soil P bioavailability, P fractions, soil pH, elec-
trical conductivity (EC), soil organic matter (SOM), and
cation exchange capacity (CEC) were also measured. This
study contributes to current debates on how biochar-derived
agro-inputs are a co-implemented solution for improving
soil health, plant nutrition and crop yield in tropical soils.
2 Materials and Methods
2.1 Soil Collection and Properties
Acidic soil was collected from Santa Bárbara municipal-
ity in northwestern Colombia (5°52′29"N, 75°33′58" W).
Soil was classified as Entisol as per USDA soil taxonomic
classification. Soil samples were collected from top layer
(0–20 cm) using methodology proposed by Muñoz et al.
(2019), thus obtaining a composite sample. Afterwards,
soil was air-dried, ground, sieved (< 2 mm) and stored. Soil
properties were: pH (5.2 ± 0.2), EC (1.3 ± 0.3 dS m−1),
SOM (3.5 ± 0.5%), CEC (13.2 ± 0.4 meq 100 g−1 soil), bio-
available P (1.2 ± 0.2 mg kg−1), sand (15%), silt (38%), clay
(47%), and texture (clay loam soil).
2.2 Production and Characterization of Raw Biochar
and Biochar‑Based Slow‑Release P Fertilizer
Corn stover was used as feedstock for CSB production.
Biomass was air-dried at room temperature (30ºC ± 2ºC)
for 48 h, then ground to pass through a 2-mm sieve and
be placed in ceramic crucibles. The crucibles were covered
with a close-fitting lid and pyrolyzed under limited oxygen
supply in a muffle furnace. The pyrolysis temperature was
raised to 500ºC for 30 min, followed by cooling at room
temperature inside a furnace. The CSB was left aerating for
24 h, then sieved (< 2 mm) and stored in airtight containers.
To produce the BBSRPF, the CSB was immersed in a satu-
rated KH2PO4 solution (400 mg P L−1) at a solid:solution
ratio of 1:40 for 48 h based on P adsorption isotherm (López
et al. 2020a) for this material to reach the highest satura-
tion (See Supplementary Information Fig. S1 and Table S1),
then the solid was separated by filtration using a 0.45 μm
membrane. Subsequently, it was washed with distilled water
(DW), dried at 60ºC and then stored.
For CSB and BBSRPF characterization, the bioavailable
P content was determined with Olsen method (Olsen-P)
(López et al., 2020b). In brief, 1.0 g of solid with 20 ml of
0.5 M NaHCO3 solution (pH 8.5) was shaken for 30 min,
then solid was removed by filtration (0.45 μm). The total P
concentration was measured by acid digestion (HCl:HNO3
3:1) at 80 °C. The P concentration was quantified using the
molybdenum blue colorimetric method with a UV–visible
8500 spectrophotometer at 800 nm. Ash content was meas-
ured as the weight of the residue after sample was heated to
700 °C in an open crucible using a muffle furnace. Super-
ficial functional groups were determined by FTIR analysis
using an infrared spectrometer (Thermo scientific Smart iTR
Nicolet iS10), which was operating in a 4000–400 cm−1
spectral range with a resolution of 4 cm−1. Specific surface
area was analyzed by BET method (López et al. 2020a) with
a surface area and pore size analyzer (Nova2200e, Quan-
tachrome Instruments). The C and N content were deter-
mined using an elemental analyzer (Elemental Analyzer,
Germany). The pH and EC were determined using 1:10 solid
to DW ratio (HQd portable multi meter – HACH ®). For
CSB, concentration of Ca, Mg, K and Na were measured
after digesting 0.25 g of sample with 10 mL of aqua regia
(HCl:HNO3) at 90 ºC for 1 h and 140 ºC for 3 h. After diges-
tion, samples were diluted with DW and filtered (0.45 μm).
Elemental concentration of elements in the filtered solution
was determined by inductively coupled plasma optical emis-
sion spectrometry (ICP-OES) (Agilent Technologies 5100
ICP-OES equipment).
2.3 Slow Phosphorus Release
To verify the slow P release from the produced BBSRPF
compared to a commercial mineral NPK fertilizer (granu-
lated fertilizer NPK 15–15-15, MONÓMEROS®, Colombia
S.A.), a mass of 100 mg P, based on P concentration of
each P source, was added in 250-mL Erlenmeyer flask and
120 mL of DW were added to it, and, afterwards, was equili-
brated for 1, 2, 3, 4, and 5 days by shaking at 100 rpm. Then,
solids were removed by filtration (0.45 μm) and P concentra-
tion was quantified as previously mentioned.
2.4 Greenhouse Pot Experiment
Certified seeds of Spinacia oleracea (Impulssemillas®,
Colombia) were used. They were germinated on peat mois-
tened with DW at 70% water holding capacity (WHC)
5 days before starting the greenhouse experiment. For
greenhouse experiment, factorial arrangements treatments
with three replicates were applied. The following treat-
ments were evaluated: i) soil with recommended mineral
13

NPK fertilization (F), ii) soil with co-application of CSB and
recommended mineral NPK fertilization (BC), and iii) soil
with co-application of the produced BBSRPF and mineral
NK fertilization (P-BC) (Table 2). Mineral NPK fertilization
was applied using commercial products, urea as N source
(MONÓMEROS®, Colombia S.A.), triple superphosphate
as P source (YARA®, Colombia S.A.), and potassium chlo-
ride as K source (MONÓMEROS®, Colombia S.A.), The
dose of BBSRPF in P-BC treatment was adjusted following
the same Olsen-P concentration used for F and BC treatment
to guarantee a similar concentration of bioavailable P. Min-
eral fertilization treatment was kept as experiment control
(Table 2), according to traditional agronomical practices.
For experimental set-up, 75 kg of soil were air-dried,
ground and sieved (< 2 mm). Then, each soil sample was
mixed with the respective treatment (Table 2). Then, the
soil samples were moistened at 70% WHC and were incu-
bated for 24 h at room temperature (30 ± 2ºC). After incu-
bation period, a soil sample was taken from each treatment
to determine initial bioavailable P concentration. Pre-incu-
bated soil samples were transferred into pots and each one
was filled with 5.0 kg of soil obtaining 15 pots per treat-
ment. In the greenhouse, the pots were randomly arranged.
Furthermore, one spinach seedling was sown in each pot.
Besides, to maintain moisture content (70% of WHC), pots
were watered daily with DW and subjected to daily obser-
vation of weight loss. The experiment was maintained for
70 days (equivalent to 1 crop production cycle). Throughout
the experiment, soil samples were taken every 10 days to
determine the kinetics of bioavailable P. After that period,
spinach plants leaves were harvested and soil samples were
taken (post-harvest soil). The 15 leaves and soil samples
obtained from all experimental units were randomly divided
into three groups with five soil and leaf samples each. Then,
the samples were mixed to obtain three composite samples
for each treatment in order to conduct post-harvest analysis.
2.5 Soil Analysis
Soil samples were used to determine bioavailable P, P frac-
tions, pH, EC, SOM, and CEC. Bioavailable P was quanti-
fied in the samples throughout the experiment and in the
post-harvest soil by Olsen-P extraction, as previously men-
tioned. In post-harvest soil samples, the inorganic P distribu-
tion was measured according to the methodology proposed
by Hedley et al. (1982). Whereby 1.0 g of the soil sample
was sequentially extracted using deionized water first and
then 0.5 M NaHCO3 solution to determine the labile P frac-
tion (Labile-P). Subsequently, the soil sample was extracted
by 1 M and 5 M NaOH solutions (NaOH1M-P and NaOH-
P5M, respectively) to quantify P bound to Fe and Al oxides
(FeAl-P). Afterwards, the soil sample was extracted by 1 M
HCl solution (HCl-P) to quantify P bound to Ca (Ca-P).
13
Journal of Soil Science and Plant Nutrition
For all the extractions, a 1:20 ratio soil:solution was used.
Finally, the residual soil was digested at 80ºC as indicated
previously to quantify occluded P (Residual-P). For each
step, the soil-solution suspensions were filtrated (0.45 μm)
and the P in the filtrate was measured as described above.
Soil pH and EC were measured in DW using 1:2 soil:DW
ratio. For determining SOM content, Walkley and Black
method was used. Finally, in order to measure CEC ammo-
nium acetate method was employed at pH 7.0 (Pansu and
Gautheyrou 2006).
2.6 Plant Analysis
By the other hand, spinach leaves were weighed to deter-
mine fresh weight biomass (FWB) in order to calculate crop
yield. Afterwards, a leaf subsample was taken (one leaf per
plant) in order to determine protein content and ascorbic
acid concentration in fresh leaf biomass using the meth-
ods proposed by Bradford (1976) and Keller and Schwager
(1977), respectively. Then, the remaining leaves were dried
at 60ºC until constant weight was reached in order to deter-
mine dry weight biomass (DWB). After that, to quantify
leaf P, N, and K concentration the dry biomass was digested
(HCl:HNO3 3:1 ratio at 80ºC). The digestate was filtrated
(0.45 μm) and the filtrate was used to quantify P, N and K in
leaf. For determining P concentration was used method as
mentioned above, while total N was determined by Kjeldhal
method. Finally, K concentration was measured by ICP-OES
as described above.
2.7 Quality Assurance and Quality Control
To assure the reliability of the results, quality assurance and
quality control, protocols were included throughout the ana-
lytical processes. All reagents were of analytical grade. For
all procedures DW (18.2 MΩ cm−1 resistivity) was used.
Synthetic multi-element solutions and samples duplicates
were included in each lot of samples for quality assurance
and evaluate reproducibility, respectively. All the material
used was washed with P-free detergent. The glassware was
soaked in acid solution (3 M HNO3) bath overnight and
washed with DW prior use.
2.8 Statistical Analysis
All data are presented as mean values and standard error of
the mean (SEM) of three replicates. Graph’s plotting was
done with Python and the statistical analysis was carried out
with Statgraphics® Centurion XVIII software. A one-way
analysis of variance (ANOVA) was used to stablish differ-
ences among the different treatments regarding soil phys-
icochemical properties, crop yield and nutritional quality.
Values of P < 0.05 were considered statistically significant,
Journal of Soil Science and Plant Nutrition

and pairwise comparisons were performed with the Tukey’s

post-test. Pearson’s correlation analysis was used to estab-
lish relationships between soil properties and crop yield and
nutritional quality. Prior to analysis, Bartlett’s test and the
Shapiro–Wilk test were applied to verify the assumptions of
homogeneity of variance and data normality, respectively.

3 Results

3.1 Characteristics of Corn Stover‑Derived Biochar

and Biochar‑Based Slow‑Release P Fertilizer

Physicochemical characteristics of CSB and BBSRPF are

shown in Table 1. The total and bioavailable P in CSB were Fig. 1  Comparison of P release behavior from biochar-based slow-
10,060 and 84 mg kg−1 (< 1% of total-P, approximately), release P fertilizer (BBSRPF) and a commercial granulated mineral
respectively. Based on bioavailable P concentration, our fertilizer. Data are expressed as mean values of two replicates ± stand-
ard error of the mean (SEM). Absence of error bars indicates that
results indicate that P bioavailability increased by more
error bars are smaller than symbols
than 1000% when CSB was equilibrated with saturated P
solution. For BBSRPF the bioavailable P concentration was
equivalent to 54% of total P. The FTIR results evidenced mineral fertilizer released 80% of the bioavailable P and the
the binding of PO34− onto the surface of CSB after P load- BBSRPF released 40% of the bioavailable P.
ing process (Fig. S2). Compared to CSB the ash content
in BBSRPF increased by 6%. The BBSRPF showed a 22% 3.2 Changes Bioavailable Soil P Concentration
reduction in surface area compared to CSB. Concentrations and Soil Properties
of Ca and Mg in CSB were elevated compared to K and Na
concentrations. Finally, pH and EC of BBSRPF was 2 units Changes in bioavailable soil P concentration across the
and 31% lower compared to CSB, respectively. Addition- experiment are shown in Fig. S3. Application of F, BC,
ally, the results of P release assay are presented in Fig. 1. and P-BC increased soil P bioavailability to a similar level
The mineral fertilizer released 60% of the total bioavail- at the beginning of the greenhouse experiment. However,
able P after 1 day of shaking, whereas the BBSRPF released after day 10 P bioavailability was significantly reduced for
20% of the bioavailable P for the same time. After 5 days, the F and BC treatments. In contrast, the soil treated with
P-BC maintained a high P bioavailability. For post-harvest
soil samples, the bioavailable P concentrations are shown
Table 1  Characteristics of corn stover derived biochar (CSB) and in Fig. 2a. The application of P-BC significantly increased
biochar-based slow-release P fertilizer (BBSRPF) by 52% the bioavailable P concentration compared to F
treatment. On the contrary, the application of BC treatment
Parameters CSB BBSRPF
significantly reduced the bioavailability of P by 22% com-
pH 10.0 ± 0.4 7.9 ± 0.3 pared to F treatment. Compared with BC treatment, in P-CB
EC (dS ­m−1) 8.40 ± 0.4 2.30 ± 0.2 treatment significantly increased by 91% the bioavailable P
Total P (mg ­kg−1) 10,060 ± 98 22 × ­103 ± 145 concentration (Table 2).
Bioavailable P (mg ­kg−1) 84.0 ± 4.2 12 × ­103 ± 251 The BC and P-BC treatments modified significantly P dis-
Ash content (%) 22.2 ± 0.2 23.5 ± 0.08 tribution in fractions compared with F treatment (Fig. 2b).
N (%) 2.2 1.1 On the one side, for F treatment P fractions were distrib-
C (%) 67.1 68.9 uted as follows: FeAl-P > Labile-P > Residual-P > Ca-P.
Surface area ­(m2 ­g−1) 380 ± 40 296 ± 55 With 60% of P accumulated in FeAl-P fraction, 25% in
Ca (mg k­ g−1) 28,492 - Labile-P fraction and 2% in Ca-P fraction. By the other
Mg (mg k­ g−1) 23,221 - side, P fraction distribution for BC treatment was FeAl-
K (mg ­kg−1) 10,742 - P > Labile-P > Ca-P > Residual-P. Where, compared to F
Na (mg ­kg−1) 90 - treatment, P accumulated in fractions FeAl-P and Labile-P
decreased to 39% and 24%, respectively; whereas the high-
EC electrical conductivity. Data are presented as mean values of three
replicates ± standard error of the mean (SEM). For N, C, Ca, Mg, K, est percentage of P in the Ca-P fraction was found for this
and Na values data are presented as values of one replicate treatment with 19%. Finally, P-BC treatment showed the

13

Fig. 2  Changes in bioavailable P concentration (a) and P fraction dis-
tribution (b) in post-harvest soil samples treated with NPK mineral
fertilizer (F), corn stover derived biochar and NPK mineral fertilizer
(BC), and biochar-based slow-release P fertilizer with mineral NK
mineral fertilizer (P-BC) after 70 days. The results represent mean
values of three replicates. Vertical bars represent the standard error
of the mean (SEM). Means followed by the same letter are not sig-
nificantly different (Tukey at p < 0.05). Labile-P: labile P. FeAl-
P: P bound to Fe and Al oxides. Ca-P: P bound to Ca. Residual-P:
occluded P
Table 2  Description of nutrient Nutrient management treatment
management treatments for
spinach cultivation Control. Soil with NPK mineral fertilizer at recommended dose (225 – 90—200 kg ­ha−1)
Soil with CSB application at dose of 3 g k­ g−1 + NPK mineral fertilizer at recommended
dose (225 – 90—200 kg ­ha−1)
Soil with BBSRPF application at dose of 3 g k­ g−1 + NK mineral fertilizer at recom-
mended dose (225—200 kg ­ha−1)
CSB corn stover-derived biochar. BBSRPF biochar-based slow-release P fertilizer. N from Urea. P from
triple superphosphate. K from potassium chloride
13
Journal of Soil Science and Plant Nutrition
greatest accumulation of P in Labile-P fraction with 36%
and decreased P content in FeAl-P fraction to 32% regard-
ing F treatment. Furthermore, P accumulated in Ca-P frac-
tion was 13% when P-BC treatment was applied. Changes
in physicochemical properties of post-harvest soil samples
are shown in Fig. 3. In general, our results showed that soils
amended with CSB and BBSRPF significantly increased by
2.7 and 1.4 units, 100 and 74%, 98 and 86%, and 206 and
230% the values of pH, EC, CEC, and SOM for BC and
P-BC treatments, respectively, compared to the soil under F
treatment. On the contrary, no significant differences were
observed for the EC, CEC and SOM values between BC and
P-BC treatments.
3.3 Plant Growth and Crop Yield
Plant growth and crop yield results after 70 days are shown
in Fig. 4a and b. The P-BC treatment significantly increased
DWB of spinach plants by 21% and 59% compared to BC
and F treatments, respectively. Additionally, based on
mineral NPK fertilization used in this study, the expected
production was 25 t ha−1. However, only P-BC treatment
achieved to reach this value, even exceeding it. Hence, this
treatment increased crop yield (t ha−1) in 34 and 17%,
compared to F and BC treatments, demonstrating a better
efficiency of P-BC treatment in nutrient uptake (Fig. 3) and
fresh biomass production in spinach plants, compared to F
and BC treatment.
3.4 Nutritional Quality
Our results show that BC treatment significantly increased
protein and ascorbic acid content in spinach leaves by 20 and
21%, respectively. However, P-BC treatment had the best per-
formance, enhancing leaf protein by 48 and ascorbic acid by
57%. In both cases, in comparison to F treatment (Fig. 4c and
d). Additionally, changes in P, N, and K concentration and
uptake by spinach plants after 70 d are shown in Fig. 5. By the
one hand, is illustrated that the application of P-BC treatment
significantly improved by 35 and 23% leaf P concentration
compared to F and BC treatments, respectively. By contrast,
no significant differences were observed between F and BC
treatments for leaf P concentration. Moreover, a significantly
Treatment code
F
BC
P-BC
Journal of Soil Science and Plant Nutrition

Fig. 3  Changes in soil pH

(a), EC (b), CEC (c), and
SOM (d) in post-harvest soil
samples treated with NPK
mineral fertilizer (F), corn
stover derived biochar and
NPK mineral fertilizer (BC),
and biochar-based slow-release
P fertilizer with NK mineral
fertilizer (P-BC) after 70 days.
EC: electrical conductivity.
CEC: cation exchange capacity.
SOM: soil organic matter. The
results represent mean values
of three replicates. Vertical bars
represent the standard error of
the mean (SEM). Means fol-
lowed by the same letter are not
significantly different (Tukey at
p < 0.05)

Fig. 4  Changes in spinach leaf

DWB (a), yield (b), protein
(c) and AA (d) content in soil
samples treated with NPK min-
eral fertilizer (F), corn stover
derived biochar and NPK min-
eral fertilizer (BC), and biochar-
based slow-release P fertilizer
with NK mineral fertilizer
(P-BC) after 70 days. DWB: dry
weight biomass. FWB: fresh
weight biomass. AA: ascorbic
acid. The results represent mean
values of three replicates. Verti-
cal bars represent the standard
error of the mean (SEM).
Means followed by the same
letter are not significantly differ-
ent (Tukey at p < 0.05). Dotted
line indicates the expected yield
for the recommended fertilizer
application rate

increase of 283 and 138% in P uptake in leaf was observed in and K uptake compared to F treatment (Fig. 5d and f). How-
P-BC treatment compared to F and BC treatments. In addition ever, P-BC treatment significantly improved the absorption of
to this, we found a strong correlation between soil bioavailable N and K compared to BC treatment (Fig. 6).
P concentration, P uptake and the concentration of P in spinach
leaves (Fig. 5a and b) (Table S1). On the other hand, the results
show that for BC and P-BC treatments N and K concentration
in leaves was significantly favored by 47 and 52% and 38 and
33%, respectively, compared to F treatment (Fig. 5c and e).
Likewise, P-BC and BC treatments significantly increased N

13
 Journal of Soil Science and Plant Nutrition

Fig. 5  Changes in leaf P, N, and K concentration (a, c, e) and uptake fertilizer (P-BC) after 70 days. The results represent mean values of
(b, d, f) by spinach plants sown in soils treated with NPK mineral three replicates. Vertical bars represent the standard error of the mean
fertilizer (F), corn stover derived biochar and NPK mineral fertilizer (SEM). Means followed by the same letter are not significantly differ-
(BC), and biochar-based slow-release P fertilizer with NK mineral ent (Tukey at p < 0.05)

Fig. 6  General scheme for the

use of biochar-based slow-
release P fertilizers (BBSRPFs)
in spinach production. EC:
electrical conductivity. SOM:
soil organic matter. CEC: cation
exchange capacity

4 Discussion agriculture. Additionally, this work contributes to current

This research supports the importance of developing
new alternatives such as BBSRPF in novel and sustain-
able approaches, regarding the relevance that P handles in
discussions on how biochar-derived agro-inputs can be an
aid to improve agronomic crop management practices of
agricultural soils in tropical areas. In general, P concen-
tration of plant-derived biochars is very low (< 1%) (Li
et al. 2020). For this reason, it is necessary complimenting

13
Journal of Soil Science and Plant Nutrition
biochar composition using P loading processes to meet
plant nutritional requirements. In this line, the results con-
firm that the P loading process to improve its bioavailable
concentration in the CSB was successful. As shown in
Fig. S1, the L-type isotherm evidences a high affinity of
the adsorbent for the adsorbate, even at low equilibrium
concentrations (López et al. 2020a). This indicates that
the CSB used is a good material for P sorption. Although
biochar is a material with a high CEC and a lower anion
exchange capacity (AEC), different mechanisms explain
the sorption of PO43− ions on CSB. For instance, in this
study the FTIR analysis demonstrated the interaction
between P and CSB functional groups (Fig. S2). Spectra
for BBSRPF showed a band at 1000–1100 cm−1 related to
the presence of PO34− (Coates 2006; Bashir et al. 2018)
and another band at 540 cm−1 related to the presence of
phosphate functional groups bonded to oxygen (O–P–O)
(Novais et al. 2018; Nardis et al. 2020). Another possible
mechanism related to P loading in CSB could have been
that PO43− ions former bridge bonds with the residual
charge of ligand bonded divalent cations (e.g., Ca2+ and
Mg2+) resulting in PO43− sorption onto the CSB. This
phenomenon may be relevant to the P loading process on
CSB for this study, as CSB has a high ratio of Ca/P to
Ca/Mg (Table 1) (Jung et al. 2016; Ahmad et al. 2018).
Additionally, increase in ash content of BBSRPF (Table 1)
suggest the formation of new mineral phases (e.g., brushite
and hydroxyapatite), possibly related to P co-precipita-
tion processes with the Ca present in the CSB (Table 1)
(Ahmad et al. 2018). Furthermore, the decrease in surface
area in the BBSRPF evidenced that the pores of the CSB
were filled or blocked P from the KH2PO4 solution (Li
et al. 2020).
For P-BC treatment, the application of BBSRPF
improved the bioavailability of P in soil. This is evidenced
by the increase in Olsen-P concentration and Labil-P frac-
tion percentage (Fig. 2a). Possibly this finding is related to
an indirect effect of BBSRPF on soil pH and the direct con-
tribution of bioavailable P (Table 1). First, the reduction of
soil acidity by increasing the pH (6.3 ± 0.2), since P has its
greatest bioavailability in soil at pH between 6 – 7 (Zhang
et al. 2016) (Fig. 3). An increase in soil pH could reduce the
formation of insoluble precipitates with Al3+, e.g., variscite
(AlPO4 2H2O), and Fe3+, e.g., sternite (FePO4 2H2O) and
vivianite ((Fe3PO4)2 8H2O), which is common in acidic
tropical soils (Zhu et al. 2018). This hypothesis is supported
by a significant reduction of FeAl-P fraction found for this
same treatment (Fig. 2b). Second, as shown in our results
(Fig. 1), P release from BBSRPF was slower compared to
mineral fertilizer, preventing the formation of insoluble P
precipitates due to the dosage of this element from BBSRPF.
This is explained by the diffusivity of P from the matrix
to the external environment (Volf and Rosolem 2020) and
desorption of P from the BBSRPF when P was adsorbed
through weak chemical bounds (Yao et al. 2013). Moreo-
ver, P can be slowly solubilized from new mineral phases
formed during the P loading process (Ahmad et al. 2018).
Because of this, the BBSRPF was able to maintain a higher
bioavailable soil P content for the 70 days of the experiment
compared to the F and BC treatments (Fig. S3).
By the other hand, the results showed that the application
of F and BC treatments significantly reduced the concen-
tration of bioavailable P compared to the P-BC treatment.
Increases in FeAl-P fraction (Fig. 2b) in soil under F treat-
ment support the theory that Fe and Al oxides retain a large
part of P added to acidic soils (pH, 4.9 ± 0.2) (Chintala et al.
2014; Zhu et al. 2018; Eduah et al. 2019) limiting the effi-
ciency of this practice to supply bioavailable P for plants
in agricultural tropical soils. This is why the bioavailable P
kinetics shows a drastic reduction in time for this treatment
(Fig. S3).
For the BC treatment, this behavior can be explained by
the high affinity of CSB for P sorption (Fig. S1), as previ-
ously discussed. Possibly the P added as triple superphos-
phate was adsorbed by the CSB incorporated into the soil,
through one of the mechanisms mentioned above, which
would explain the rapid reduction in P bioavailability
(Fig. S3). This resulted in a reduction in the Labil-P frac-
tion (Fig. 2b) (Xu et al. 2014; Li et al. 2020). This, could
later lead to the release of P adsorbed by the CSB to soil
solution, depending on the concentration gradients of this
element, implying a residual effect that was not evaluated in
this study. However, this could generate a pool of P bioavail-
able in the long term. Besides, P can not only be retained in
CSB in the BC treatment, but also in SOM (Xu et al. 2014);
therefore, it is possible that a higher increase in SOM con-
tent in this treatment (Fig. 3d) may be related to a lower P
bioavailability. Additionally, in BC treatment, soil pH was
slightly alkaline (7.8 ± 0.2) (Fig. 3a) which could promote
formation of Ca-P insoluble precipitates (Zhu et al. 2018)
decreasing P bioavailability, which can be evidenced in Ca-P
fraction increase for this treatment (Fig. 2b).
These findings are in agreement with other studies where
it has been determined that the application of BBSRPF can
be more efficient for improving P bioavailability in soils
than traditional fertilizers and co-application of CSB itself
and mineral P fertilizers (Carneiro et al. 2021; Lustosa
Filho et al. 2020; Pogorzelski et al. 2020). The results of
this study imply that the effect of CSB on P bioavailability
would depend on the affinity of the material for this ele-
ment. Thus, it is necessary to previously saturate the CSB
with P so that it does not adsorb the P present in the soil
solution and instead releases the P retained in the matrix of
the material.
Besides the positive effects of P-BC treatment and nega-
tive effects of F and BC treatments on P soil bioavailability
13

at 70 d, effects of these treatments on other physicochemical
properties of the soil were also found. For instance, results
showed increases in pH, EC, CEC, and SOM for BC and
P-BC treatments compared to F treatment. Similar results
were found in other studies, where the application of differ-
ent types of biochars improved physicochemical properties
of soil, increasing its quality (Chew et al. 2020). Further-
more, no differences between BC and P-BC treatments were
found regarding EC, CEC, and SOM, suggesting that the
CSB P loading process did not decrease its capability to
improve soil health.
Changes in pH were significantly different between treat-
ments (Fig. 3a). The increase in pH of soil treated with CSB
and BBSRPF is due to the liming effect of these materials
(Shi et al. 2017). However, the application of CSB increased
the soil pH value 1.1 units compared to the soil treated with
BBSRPF (Fig. 3a). This is related to a lower pH and EC
value of the BBSRPF compared to CSB (Table. 1), due to
the remotion of some basic compounds by the agitation and
washing (López et al. 2020a) in the process of preparation
of BBSRPF. The increase in CEC (Fig. 3c) is explained by
the increment in soil pH. Which enhances the amount of
negative soil surface charges, in addition to the contribu-
tion of surface functional groups (Fig. S2) (Shi et al. 2017).
Furthermore, the application of CSB and BBSRPF increased
SOM (Fig. 3d) due to the organic carbon inputs (Shi et al.,
2020) (Table 1). Besides, increases in soil EC in BC and
P-BC treatments (Fig. 3b) are explained as a result of releas-
ing of cations and anions from the materials into soil solu-
tion (Khan et al. 2017). These results demonstrate that the
BBSRPF promotes the “biochar effect” (Chew et al. 2020),
which improves both soil health and bioavailability of min-
eral nutrient, such as P.
As a result of the positive incidence that BBSRPF has
on soil health improvement, crop production and nutri-
tional value are also enhanced. Which is in accordance
with our results, since application of BC and P-BC treat-
ments improved soil physicochemical properties (Fig. 3).
Particularly, P-BC treatment showed the best results for all
the variables determined in this study, which are consistent
with better crop productivity. For instance, DWB of spinach
plants was significantly increased when both BBSRPF and
CSB were applied (Fig. 4a). As similarly found by Younis
et al. (2016) and Li et al. (2016) regarding the increase in
spinach DWB with application of biochar produced from
cotton stalks and maize straw, respectively. However, P-BC
treatment showed the best performance in DWB increase
compared to BC and F treatments, reaffirming the superior-
ity of BBSRPF, as has been also found in previous stud-
ies (Ahmad et al. 2018). Additionally, crop yield was also
higher with P-BC treatment while F treatment had the poor-
est performance for improving spinach yield (Fig. 4b), which
corroborates the low effectiveness of conventional fertilizers
13
Journal of Soil Science and Plant Nutrition
when added to soil. Since these products are usually either
limited by precipitation processes with soil components or
leached easily (Li et al. 2020), reducing nutrients bioavail-
ability for plants and, consequently, causing their lower
growth and development. Furthermore, these results are in
line with other studies in which the application of organic-
based amendments improve soil nutrient bioavailability for
crops, compared to mineral NPK fertilizers (Klammsteiner
et al. 2020).
Moreover, the application of BBSRPF also promoted a
better nutritional quality of spinach regarding ascorbic acid,
protein (Fig. 4c and d) and macronutrients (N, P and K)
content in leaf (Fig. 5a,c and e), which were significantly
enhanced with P-BC treatment. By the one hand, ascorbic
acid is one of the most important nutritional quality factors
in many crops, since performs many biological functions in
the human body. Mainly, regarding its antioxidant activity
which can help reducing the risk of cardiovascular diseases
and some cancer types (Citak and Sonmez 2010; Mozafar
1993; Salehi et al. 2019). This substance is provided mostly
by plants (Wang et al. 2013), which emphasizes the need of
enhances nutritive quality of crops (Gallie 2013). On the
other hand, due to the emerging and growing awareness of
negative effects of animal proteins sources on human health,
there is an increasing interest on vegetable proteins, which,
in turn, have been demonstrated to benefit cardiovascular
health and lipid metabolism while providing a wide profile
of amino acids of importance for human nutrition. Addition-
ally, lower economic and environmental costs of vegetable
proteins are also some of the advantages of this alterna-
tive source (Marti-Quijal et al. 2019). As mentioned, the
increase of ascorbic acid and protein in plants grown in soils
treated with P-CB was higher than that of plants planted
in soils under BC and F treatments. Previously, some stud-
ies have showed that biochemical attributes such as protein
and ascorbic acid content in spinach leaf can be strongly
improved with biochar application. For instance, Khan et al.
(2017) found that miscanthus biochar improved protein and
ascorbic acid of spinach up to 77 and 251%, respectively.
Similarly, Turan (2020) also observed that the improvement
of protein content at leaf level was promoted when biochar-
based amendments were applied to spinach crops. Improved
plant nutritional value under P-BC treatment correlated with
changes in soil physicochemical properties (Table S3).
Likewise, macronutrients (N, P, and K) content in spin-
ach leaves was also shown to be mainly enhanced with the
application of BBSRPF (Fig. 5). The superior performance
of this material to increase P concentration and uptake is
related to the increase in soil P bioavailability (Fig. 2a)
(Table S2). Since, compared to the dissolution of mineral P
fertilizer, P adsorbed on biochar matrix would be released
according to the adsorption–desorption equilibrium between
plant roots and soil. In addition, BBSRPF had a low bulk
Journal of Soil Science and Plant Nutrition
density compared to mineral fertilizer (Fig. S4). Therefore,
it may be better distributed in the soil, generating a higher
possibility of contact between roots and BBSRPF particles,
which act as a hot-spot of bioavailable soil P source (Li
et al. 2020). Which is supported by the correlation found
among these parameters (Table S2). Additionally, N and K
content were increased in P-BC and BC treatments. Despite
a possible increase in soil K bioavailability due to the use of
KH2PO4 solution for the CSB loading process, and for K
content in CSB, leaf K concentration was not different for
BC and P-BC treatments. Possibly this is explained by the
fact that both treatments supplied the bioavailability of this
nutrient in the soil, necessary for the spinach plant, since
nutrient uptake by plants depends not only on the concen-
tration of the element in soil solution, but also on the nutri-
tional requirement of the plant. Therefore, no deficiencies or
increases in this parameter were observed. However, in the
long term, the BBSRPF produced could represent a reserve
not only of P but also of bioavailable K. This should be
evaluated in trials with several crop cycles, to determine
the possible advantages of BBSRPF in reducing the appli-
cation of mineral K fertilizers. Even though no significant
differences were found regarding N and K content at leaf
level between these treatments, the uptake efficiency of
these nutrients was indeed greatly increased by BBSRPF
(Fig. 5). Since BBSRPF provides nutrients and maintains
a greater bioavailability of them in rhizosphere of spinach
plants, improving their uptake (Table S3). This shows that
the ability of BBSRPFs to slowly dose nutrients and main-
tain their bioavailability over time is an important factor on
soil fertility management (Khan et al. 2017).
5 Conclusion
This study confirms that corn stover-derived biochar (CSB)
loaded with P can act as a biochar-based slow-release P fer-
tilizer (BBSRPF), which can replace traditional P fertilizers,
in order to meet the requirements of this nutrient in spinach
crops in acidic soils. Our results support the hypothesis that
BBSRPFs can improve the nutritional quality of spinach
and increase yield, because fertility conditions of soils are
improved by its application. In this context, it can be stated
that BBSRPFs application to agronomic crops could work
as a P fertilization strategy compared to commercial mineral
P fertilizers. From an environmental point of view, this type
of fertilizer represents a viable alternative for the sustain-
able management of crops fertilization schemes. Since slow
liberation mechanisms of nutrients from BBSRPFs improve
nutrient use by plants and may prevent nutrient leaching
and groundwater contamination. However, field-scale trials
should be developed to test the effectiveness and applicabil-
ity of this form of P fertilization at a greater scale.
Supplementary Information The online version contains supplemen-
tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 42729-0​ 21-0​ 0583-0.
Acknowledgements The authors thank the laboratory of the Institución
Universitaria Colegio Mayor de Antioquia for its technical support and
Semillero de Investigación en Ciencias Ambientales—SICA. Further-
more, Julián E. López thanks Universidad de Medellín for the scholar-
ship [No. 321316/427405].
Author Contribution Maicol Torres and Evelyn Becerra-Agudelo: soil
samples collection, methodology validation, assays development, data
collection, data analysis, and writing the first draft of the paper.
Cindy Sepúlveda-Cadavid and Jhon H. Romero: conceptualization,
experimental design, data analysis, review, and editing of the paper.
Julián E. López: conceptualization, funding acquisition, experimen-
tal design, data analysis, supervision, review, and editing of the paper.
Funding This study was funded by Institución Universitaria Colegio
Mayor de Antioquia and Universidad de Medellín.
Data Availability The datasets generated during the current study are
available from the corresponding author on reasonable request.
Code Availability Not applicable.
Declarations
Conflict of Interest The authors declare no competing interests.
References
Ahmad M, Usman ARA, Al-Faraj AS, Ahmad M, Sallam A, Al-Wabel
MI (2018) Phosphorus-loaded biochar changes soil heavy metals
availability and uptake potential of maize (Zea mays L.) plants.
Chemosphere 194:327–339. https://​doi.​org/​10.​1016/j.​chemo​
sphere.​2017.​11.​156
Aiking H (2011) Future protein supply. Trends Food Sci Technol
22:112–120. https://​doi.​org/​10.​1016/J.​TIFS.​2010.​04.​005
Bashir S, Hussain Q, Shaaban M, Hu H (2018) Efficiency and surface
characterization of different plant derived biochar for cadmium
(Cd) mobility, bioaccessibility and bioavailability to Chinese cab-
bage in highly contaminated soil. Chemosphere 211:632–639.
https://​doi.​org/​10.​1016/j.​chemo​sphere.​2018.​07.​168
Bergquist SÅM, Gertsson UE, Olsson ME (2006) Influence of growth
stage and postharvest storage on ascorbic acid and carotenoid con-
tent and visual quality of baby spinach (Spinacia oleracea L.).
J Sci Food Agric 86:346–355. https://​doi.​org/​10.​1002/​jsfa.​2373
Bojacá CR, Wyckhuys KAG, Gil R, Jiménez J, Schrevens E (2010)
Sustainability aspects of vegetable production in the peri-urban
environment of Bogotá, Colombia. Int J Sustain Dev World Ecol
17:487–498. https://​doi.​org/​10.​1080/​13504​509.​2010.​516890
Bolan NS, Adriano DC, Curtin D (2003) Soil acidification and liming
interactions with nutrientand heavy metal transformationand bio-
availability. Adv Agron 78:215–272
Bouwman L, Goldewijk KK, Van Der Hoek KW, Beusen AHW, Van
Vuuren DP, Willems J, Rufino MC, Stehfest E (2013) Exploring
global changes in nitrogen and phosphorus cycles in agriculture
induced by livestock production over the 1900–2050 period. Proc
Natl Acad Sci U S A 110:20882–20887. https://​doi.​org/​10.​1073/​
pnas.​10128​78108
Bradford M (1976) A Rapid and Sensitive Method for the Quantita-
tion of Microgram Quantities of Protein Utilizing the Principle of
13

Protein-Dye Binding. Anal Biochem 72:248–254. https://​doi.​org/​
10.​1006/​abio.​1976.​9999
Carvalho FAL, Pateiro M, Domínguez R, Barba-Orellana S, Mattar J,
Rimac Brnčić S, Barba FJ, Lorenzo JM (2019) Replacement of
meat by spinach on physicochemical and nutritional properties of
chicken burgers. J Food Process Preserv 43:e13935. https://​doi.​
org/​10.​1111/​jfpp.​13935
Casierra-Posada F, Briceño-Pinzón ID, Carreño-Patiño JA (2021)
Tolerance of Spinach ( Spinacia oleracea ) Plants to Partial
Defoliation. Gesunde Pflanz 2021:1–8. https://​doi.​org/​10.​1007/​
S10343-​021-​00561-7
Chew J, Zhu L, Nielsen S et al (2020) Biochar-based fertilizer: Super-
charging root membrane potential and biomass yield of rice. Sci
Total Environ 713:136431. https://​doi.​org/​10.​1016/j.​scito​tenv.​
2019.​136431
Chintala R, Schumacher TE, McDonald LM, Clay DE, Malo DD,
Papiernik SK, Clay SA, Julson JL (2014) Phosphorus Sorp-
tion and Availability from Biochars and Soil/Biochar Mixtures.
CLEAN - Soil, Air, Water 42:626–634. https://​doi.​org/​10.​1002/​
clen.​20130​0089
Citak S, Sonmez S (2010) Effects of conventional and organic fertili-
zation on spinach (Spinacea oleracea L.) growth, yield, vitamin
C and nitrate concentration during two successive seasons. Sci
Hortic (amsterdam) 126:415–420. https://​doi.​org/​10.​1016/j.​scien​
ta.​2010.​08.​010
Coates J (2006) Interpretation of Infrared Spectra, A Practical
Approach. Encyclopedia of Analytical Chemistry. John Wiley &
Sons Ltd, Chichester
da Carneiro JS, S, Ribeiro ICA, Nardis BO, Barbosa CF, Lustosa Filho
JF, Melo LCA, (2021) Long-term effect of biochar-based fertiliz-
ers application in tropical soil: Agronomic efficiency and phos-
phorus availability. Sci Total Environ 760:143955. https://d​ oi.o​ rg/​
10.​1016/J.​SCITO​TENV.​2020.​143955
Eduah JO, Nartey EK, Abekoe MK, Breuning-Madsen H, Andersen
MN (2019) Phosphorus retention and availability in three contrast-
ing soils amended with rice husk and corn cob biochar at varying
pyrolysis temperatures. Geoderma 341:10–17. https://​doi.​org/​10.​
1016/j.​geode​rma.​2019.​01.​016
Fan D, Kandasamy S, Hodges DM, Critchley AT, Prithiviraj B (2014)
Pre-harvest treatment of spinach with Ascophyllum nodosum
extract improves post-harvest storage and quality. Sci Hortic
(amsterdam) 170:70–74. https://​doi.​org/​10.​1016/j.​scien​ta.​2014.​
02.​038
Gallego JL, Olivero-Verbel J (2021) Cytogenetic toxicity from pes-
ticide and trace element mixtures in soils used for conventional
and organic crops of Allium cepa L. Environ Pollut 276:116558.
https://​doi.​org/​10.​1016/j.​envpol.​2021.​116558
Gallie DR (2013) L-Ascorbic Acid: A Multifunctional Molecule
Supporting Plant Growth and Development. Scientifica (Cairo)
2013:735964. https://​doi.​org/​10.​1155/​2013/​795964
Hedley MJ, Stewart JWB, Chauhan BS (1982) Changes in Inorganic
and Organic Soil Phosphorus Fractions Induced by Cultivation
Practices and by Laboratory Incubations. Soil Sci Soc Am J
46:970–976. https://d​ oi.o​ rg/1​ 0.2​ 136/s​ ssaj1​ 982.0​ 36159​ 95004​ 6000​
50017x
Jung KW, Jeong TU, Kang HJ, Ahn KH (2016) Characteristics of bio-
char derived from marine macroalgae and fabrication of granular
biochar by entrapment in calcium-alginate beads for phosphate
removal from aqueous solution. Bioresour Technol 211:108–116.
https://​doi.​org/​10.​1016/j.​biort​ech.​2016.​03.​066
Keller T, Schwager H (1977) Air pollution and ascorbic acid. For
Pathol 7:338–350. https://​doi.​org/​10.​1111/j.​1439-​0329.​1977.​
tb006​03.x
Khan WD, Ramzani PMA, Anjum S, Abbas F, Iqbal M, Yasar A,
Ihsan MZ, Anwar MN, Baqar M, Tauqeer HM, Virk ZA, Khan
SA (2017) Potential of miscanthus biochar to improve sandy soil
13
Journal of Soil Science and Plant Nutrition
health, in situ nickel immobilization in soil and nutritional qual-
ity of spinach. Chemosphere 185:1144–1156. https://​doi.​org/​10.​
1016/j.​chemo​sphere.​2017.​07.​097
Klammsteiner T, Turan V, Juárez MF-D, Oberegger S, Insam H (2020)
Suitability of Black Soldier Fly Frass as Soil Amendment and
Implication for Organic Waste Hygienization. Agron 1578:10–
1578. https://​doi.​org/​10.​3390/​AGRON​OMY10​101578
Lander EM, Wertheim BC, Koch SM, Chen Z, Hsu CH, Thomson CA
(2016) Vegetable protein intake is associated with lower gallblad-
der disease risk: Findings from the Women’s Health Initiative
prospective cohort. Prev Med (baltim) 88:20–26. https://​doi.​org/​
10.​1016/j.​ypmed.​2016.​03.​016
Lara OA, Amorós A, Tapia ML, Escalona VH (2021) Effect of a pho-
toselective filter on the yield and postharvest quality of “Viroflay”
baby spinach (Spinacia oleracea L.) leaves cultivated in a hydro-
ponic system. Sci Hortic (amsterdam) 277:109804. https://d​ oi.o​ rg/​
10.​1016/J.​SCIEN​TA.​2020.​109804
Li ZF, Wang Q, Zhang WJ, Du ZL, He XH, Zhang QZ (2016) Contri-
butions of Nutrients in Biochar to Increase Spinach Production:
A Pot Experiment. Commun Soil Sci Plant Anal 47:2003–2007.
https://​doi.​org/​10.​1080/​00103​624.​2016.​12250​78
Li H, Li Y, Xu Y, Lu X (2020) Biochar phosphorus fertilizer effects on
soil phosphorus availability. Chemosphere 244:125471. https://​
doi.​org/​10.​1016/j.​chemo​sphere.​2019.​125471
Lin W, Lin M, Zhou H, Wu H, Li Z, Lin W (2019) The effects of
chemical and organic fertilizer usage on rhizosphere soil in tea
orchards. PLoS ONE 14:0217018. https://​doi.​org/​10.​1371/​journ​
al.​pone.​02170​18
López JE, Builes S, Heredia Salgado MA, Tarelho LAC, Arroyave
C, Aristizábal A, Chavez E (2020a) Adsorption of Cadmium
Using Biochars Produced from Agro-Residues. J Phys Chem C
124:14592–14602. https://​doi.​org/​10.​1021/​acs.​jpcc.​0c022​16
López JE, Gallego JL, Vargas-Ruiz A, Peña-Mosquera AL, Zapata-
Zapata AD, López-Sánchez IJ, Botero-Botero LR (2020b) Asper-
gillus tubingensis and Talaromyces islandicus Solubilize Rock
Phosphate Under Saline and Fungicide Stress and Improve Zea
mays Growth and Phosphorus Nutrition. J Soil Sci Plant Nutr
20:2490–2501. https://​doi.​org/​10.​1007/​s42729-​020-​00315-w
Lustosa Filho JF, da Carneiro JS, Barbosa CF, de Lima KP, Leite do
A, Melo LCA (2020) Aging of biochar-based fertilizers in soil:
Effects on phosphorus pools and availability to Urochloa bri-
zantha grass. Sci Total Environ 709:136028. https://​doi.​org/​10.​
1016/J.​SCITO​TENV.​2019.​136028
Marti-Quijal FJ, Zamuz S, Tomašević I, Gómez B, Rocchetti G, Lucini
L, Remize F, Barba FJ, Lorenzo JM (2019) Influence of differ-
ent sources of vegetable, whey and microalgae proteins on the
physicochemical properties and amino acid profile of fresh pork
sausages. LWT 110:316–323. https://​doi.​org/​10.​1016/j.​lwt.​2019.​
04.​097
Mozafar A (1993) Nitrogen fertilizers and the amount of vitamins in
plants: A review. J Plant Nutr 16:2479–2506
Muñoz Romero JH, Sepúlveda Cadavid CA, Cortés N, López Correa
JE, Correa Estrada JD (2019) Inactivation of Fusarium oxyspo-
rum Conidia in Soil with Gaseous Ozone-Preliminary Studies.
Ozone Sci Eng 42:36–42. https://d​ oi.o​ rg/1​ 0.1​ 080/0​ 19195​ 12.2​ 019.​
16088​10
Naeem I, Masood N, Turan V, Iqbal M (2021) Prospective usage of
magnesium potassium phosphate cement combined with Bougain-
villea alba derived biochar to reduce Pb bioavailability in soil
and its uptake by Spinacia oleracea L. Ecotoxicol Environ Saf
208:111723. https://​doi.​org/​10.​1016/J.​ECOENV.​2020.​111723
Nardis BO, da Silva S, Carneiro J, de Souza IMG, de Barros RG,
Azevedo Melo LC (2020) Phosphorus recovery using magnesium-
enriched biochar and its potential use as fertilizer. Arch Agron
Soil Sci 03650340(2020):1771699. https://d​ oi.o​ rg/1​ 0.1​ 080/0​ 3650​
340.​2020.​17716​99
Journal of Soil Science and Plant Nutrition
Novais SV, Zenero MDO, Tronto J, Conz RF, Cerri CEP (2018) Poultry
manure and sugarcane straw biochars modified with M ­ gCl2 for
phosphorus adsorption. J Environ Manage 214:36–44. https://d​ oi.​
org/​10.​1016/j.​jenvm​an.​2018.​02.​088
Pansu M, Gautheyrou J (2006) Handbook of Soil Analysis. Springer
Berlin Heidelberg, Berlin
Pogorzelski D, Filho JFL, Matias PC, Santos WO, Vergütz L, Melo
LCA (2020) Biochar as composite of phosphate fertilizer: Char-
acterization and agronomic effectiveness. Sci Total Environ
743:140604. https://d​ oi.o​ rg/1​ 0.1​ 016/J.S
​ CITOT​ ENV.2​ 020.1​ 40604
Roberts JL, Moreau R (2016) Functional properties of spinach (Spi-
nacia oleracea L.) phytochemicals and bioactives. Food Funct
7:3337–3353
Sabaté J, Sranacharoenpong K, Harwatt H, Wien M, Soret S (2015)
The environmental cost of protein food choices. Public Health
Nutr 18:2067–2073. https://d​ oi.o​ rg/1​ 0.1​ 017/S
​ 13689​ 80014​ 00237​ 7
Salehi B, Tumer TB, Ozleyen A et al (2019) Plants of the genus Spina-
cia: From bioactive molecules to food and phytopharmacological
applications. Trends Food Sci Technol 88:260–273
Shi R, Hong Z, Li J, Jiang J, Baquy MA-A, Xu R, Qian W (2017)
Mechanisms for Increasing the pH Buffering Capacity of an
Acidic Ultisol by Crop Residue-Derived Biochars. J Agric Food
Chem 65:8111–8119. https://​doi.​org/​10.​1021/​acs.​jafc.​7b022​66
Shi RY, Ni N, Nkoh JN, Dong Y, Zhao WR, Pan XY, Li JY, Xu RK,
Qian W (2020) Biochar retards Al toxicity to maize (Zea mays L.)
during soil acidification: The effects and mechanisms. Sci Total
Environ 719:137448. https://​doi.​org/​10.​1016/j.​scito​tenv.​2020.​
137448
Turan V (2019) Confident performance of chitosan and pistachio
shell biochar on reducing Ni bioavailability in soil and plant
plus improved the soil enzymatic activities, antioxidant defense
system and nutritional quality of lettuce. Ecotoxicol Environ Saf
183:109594. https://​doi.​org/​10.​1016/J.​ECOENV.​2019.​109594
Turan V (2020) Potential of pistachio shell biochar and dicalcium phos-
phate combination to reduce Pb speciation in spinach, improved
soil enzymatic activities, plant nutritional quality, and antioxi-
dant defense system. Chemosphere 245:125611. https://​doi.​org/​
10.​1016/j.​chemo​sphere.​2019.​125611
Volf MR, Rosolem CA (2020) Soil P Diffusion and Availability Modi-
fied by Controlled-Release P Fertilizers. J Soil Sci Plant Nutr
21:162–172. https://​doi.​org/​10.​1007/​s42729-​020-​00350-7
Wang J, Zhang Z, Huang R (2013) Regulation of ascorbic acid synthe-
sis in plants. Plant Signal Behav 8:e24536
Xu G, Sun J, Shao H, Chang SX (2014) Biochar had effects on phos-
phorus sorption and desorption in three soils with differing acid-
ity. Ecol Eng 62:54–60. https://​doi.​org/​10.​1016/j.​ecole​ng.​2013.​
10.​027
Yao Y, Gao B, Chen J, Yang L (2013) Engineered Biochar Reclaiming
Phosphate from Aqueous Solutions: Mechanisms and Potential
Application as a Slow-Release Fertilizer. Environ Sci Technol
47:8700–8708. https://​doi.​org/​10.​1021/​ES401​2977
Younis U, Malik SA, Rizwan M, Qayyum MF, Ok YS, Shah MHR,
Rehman RA, Ahmad N (2016) Biochar enhances the cadmium
tolerance in spinach (Spinacia oleracea) through modification of
Cd uptake and physiological and biochemical attributes. Envi-
ron Sci Pollut Res 23:21385–21394. https://​doi.​org/​10.​1007/​
s11356-​016-​7344-3
Zhang H, Chen C, Gray EM, Boyd SE, Yang H, Zhang D (2016) Roles
of biochar in improving phosphorus availability in soils: A phos-
phate adsorbent and a source of available phosphorus. Geoderma
276:1–6. https://​doi.​org/​10.​1016/j.​geode​rma.​2016.​04.​020
Zhu J, Li M, Whelan M (2018) Phosphorus activators contribute to
legacy phosphorus availability in agricultural soils: A review. Sci
Total Environ 612:522–537. https://​doi.​org/​10.​1016/j.​scito​tenv.​
2017.​08.​095
Zubair M, Adnan Ramzani PM, Rasool B, Khan MA, ur-Rahman M,
Akhtar I, Turan V, Tauqeer HM, Farhad M, Khan SA, Iqbal J,
Iqbal M, (2021) Efficacy of chitosan-coated textile waste biochar
applied to Cd-polluted soil for reducing Cd mobility in soil and its
distribution in moringa (Moringa oleifera L.). J Environ Manage
284:112047. https://​doi.​org/​10.​1016/J.​JENVM​AN.​2021.​112047
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