Science of the Total Environment 851 (2022) 158141

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Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

A field study on using soybean waste-derived superabsorbent hydrogel to

enhance growth of vegetables
Jingling Zhu a,b, Fadhlina Suhaimi c, Jing Ying Lim a, Zhengyang Gao a,b, Sanjay Swarup a,d,e, Chiang Shiong Loh a,d,
⁎
Jun Li b, Choon Nam Ong f, Wee Kee Tan a,
a
NUS Environmental Research Institute, National University of Singapore, 5A Engineering Drive 1, T-Lab, #02-01, Singapore 117411, Singapore
b
Department of Biomedical Engineering, National University of Singapore, 7 Engineering Drive 1, Singapore 117574, Singapore
c
Singapore Food Agency, 52 Jurong Gateway Road, #14-01, Singapore 608550, Singapore
d
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore
e
Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore
f
Saw Swee Hock School of Public Health, National University of Singapore, 12 Science Drive 2, Singapore 117549, Singapore

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• A field assessment of okara-derived super-

absorbent hydrogel on leafy vegetables
• The hydrogel enhances vegetable growth
and increases water use efficiency.
• Higher P and K retained in soil with the
hydrogel suggests better fertilizer usage.
• Statistical modelling of field data provides
useful information to crop growers.
• Such waste-to-resource approach offers
plant-based soil supplement for sustain-
able farming.

A R T I C L E I N F O A B S T R A C T

Editor: Daniel CW Tsang Food security is critical and has become a global concern with many of our basic food crops growing in areas with high
drought risk. To improve soil water holding capacity, hydrogels are a promising solution. However, the current ones
Keywords: are mostly derived from petroleum products and are environmental unsustainable. In this study, the main objective
Superabsorbent polymers is to determine if bio-based hydrogel can help in the growth of leafy vegetables while minimizing water use under
Field-testing
field conditions. To achieve this, we developed an okara-derived hydrogel (Ok-PAA; OP) from by-products of bean
Sustainable crop production
Nutrient-saving
curd and soybean milk production. We incorporated OP into soil and assessed the growth performance of leafy vege-
Climate change tables. We observed that vegetables grown with 0.2% (w/v) OP in soil with a watering frequency of 7 times per week
Water use efficiency resulted in >60 % and 35 % yield increase for the common Asian leafy vegetables, choy sum (CS) and pak choi (PC),
respectively, as compared to without hydrogel supplementation. Both vegetables produced larger leaf areas (20–40 %
increment) in the presence of the hydrogel as compared to those without. In addition, with OP amendment, the irriga-
tion water use efficiency improved >60 % and 30 % for CS and PC, respectively. It is estimated that with the use of the
hydrogel, a reduction in watering frequency from 21 times to 7 times per week could be achieved, and based on a per
hectare estimation, this would result in 196,000 L of water saving per crop cycle. Statistical analysis and modelling fur-
ther confirmed vegetables grown with 0.2 % (w/v) OP and with a watering frequency of 7 times per week showed the

⁎ Corresponding author.
E-mail address: eritwk@nus.edu.sg (W.K. Tan).

http://dx.doi.org/10.1016/j.scitotenv.2022.158141
Received 15 May 2022; Received in revised form 16 August 2022; Accepted 16 August 2022
Available online 19 August 2022
0048-9697/© 2022 Elsevier B.V. All rights reserved.
J. Zhu et al.
best growth performance and water use efficiency. Such a waste-to-resource approach offers a plant-based soil supple-
ment for crop growers, contributes to waste valorization, and enhances the growth of plants especially under water-
limited conditions.
1. Introduction
Climate change has increasingly become a topic of global concern. The
speed at which climate and weather patterns are changing might result in
our current agricultural systems being less stable, thus threatening a secure
food future (O'Brien et al., 2021). This instability adds to the existing chal-
lenges to feed the expected world population of 8–10 billion by 2050 (FAO,
2009; Paliyath et al., 2019). To meet this demand, it has been estimated
that food production will need to grow by 70 % (Shinde et al., 2019). How-
ever, crop harvest is severely affected under water deficit conditions, and
many of our basic food crops are grown in areas with high drought risk
(Barnabás et al., 2008; Fahad et al., 2017; Pennisi, 2008; Vatter et al.,
2019). Since water is crucial for plant growth, agriculture is water-
intensive, and consumes about 70 % of the world's freshwater, mainly
through unsustainable rates of groundwater abstraction (de Graaf et al.,
2019; Grafton et al., 2018; Rodell et al., 2018). With climate change and
worsening drought conditions predicted over the next decades, sourcing
for alternative approaches to meet the water demand for sustainable crop
production become an urgent task.
Taking into consideration that the bulk of the irrigated water is usually
evaporated or leached out rather than being taken up by the plants
(Bourzac, 2013), developing methods that can retain the water in the soil
for longer duration would be one possible approach to sustain the growth
of the plants, especially under water-limited conditions. While the use of
sprinkler or drip irrigation systems could increase the irrigation efficiency
and keep the plants hydrated, these might be more suitable for higher
value cash crops due to their high capital and maintenance cost (Bourzac,
2013; de Graaf et al., 2019; Locascio, 2005). Besides working on the irriga-
tion efficiency, the use of high efficiency water-absorbent materials, such as
superabsorbent polymers (SAP) to improve on the soil capability to retain
water would be a more direct approach. SAP, or more commonly known
as hydrogels, are able to imbibe water that is hundreds of times higher
than their own weight and have gained much attractions (Ahmed, 2015;
Qi et al., 2021; Venkatachalam and Kaliappa, 2021). In fact, many of
these polymers have been applied in agronomy. Synthetic SAP (for exam-
ples, Stockosorb Agro, Luquasorb, STOCKSORB 660 medium, SaturAid®)
had been tested on young citrus plants, tree seedlings (examples: Eucalyptus
grandis, Pinus caribaea, Araucaria cunninghamii, Terminalia superba),
common sage (Salvia officinalis), winter wheat (Triticum aestivum), white
lupin (Lupinus albus) and Italian ryegrass (Lolium multiflorum) (Agaba
et al., 2010; Arbona et al., 2005; Paradelo et al., 2019; Savi et al., 2014).
Nevertheless, based on our knowledge, field study on green leafy vegeta-
bles using SAP has rarely been reported.
With the increased awareness on environmental sustainability and
waste valorization, rather than relying on the use of synthetic petroleum-
based materials, reusing waste materials, such as flax yarn (Wu et al.,
2012) mulberry branches (Zhang et al., 2014), hemicelluloses lye (Liu
et al., 2019), crop straw (Li and Chen, 2020) and white cabbage (Zhang
et al., 2021) had been reported for the production of natural superabsor-
bent hydrogels in recent years. Through a waste-to-resource approach, we
had previously reported on the development of a soybean residue
(okara)-based hydrogel and had shown the efficacy of this hydrogel as a
soil supplement in potted plants under indoor settings where the environ-
mental conditions are better-controlled (Li et al., 2019; Zhu et al., 2020a;
Zhu et al., 2020b). Under field conditions, however, plants are subjected
to more challenging growth conditions, with ambient temperature, light
intensity and relative humidity fluctuations depending on the site weather
conditions. The vegetables are also grown together on the ground in raised
beds similar to commercial vegetable farming rather than growing
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Science of the Total Environment 851 (2022) 158141
individually in pots when they were grown under indoor conditions.
Information on the suitability of such okara-based hydrogel application
on a larger scale plant performance, with plants grown in raised beds
under less predictable field conditions is lacking, especially for green
leafy vegetables. It will thus be interesting to determine if OP can still en-
hance the performance of plants under more challenging growth situations.
In this study, we first modified the hydrogel synthesis for larger scale
production so that sufficient amount of okara-derived hydrogel (Ok-PAA;
OP) can be produced for field assessment. We hypothesize that OP, when
added to the soil in appropriate amount, can help in the growth of field-
grown leafy vegetables while minimizing water use. To achieve this, we
selected yield in terms of shoot fresh weights and leaf area, as well as irriga-
tion WUE as the indicators for plant growth assessment. We determined the
effect of different concentrations of OP added to soil, together with various
watering frequencies applied to the plants, on the growth performance of
two commonly consumed Asian green leafy vegetables, choy sum (CS)
and pak choi (PC), under field conditions. Since the okara-derived hydrogel
improves the water holding and retention properties of soil (Zhu et al.,
2022), we further hypothesize that OP helps in nutrient retention. Another
important aim of the current field study is to provide information to the
crop growers on the application of the okara-derived hydrogel based on
yield and water-use prediction through statistical modelling.
2. Materials and methods
2.1. Synthesis of okara-derived hydrogel (hereinafter referred to as Ok-PAA; OP)
Okara-derived hydrogel (Ok-PAA; OP) was synthesized according to Zhu
et al. (2020b, 2022). A similar free radical polymerization technique was em-
ployed to synthesize okara-poly(acrylic acid) (Ok-PAA) hydrogels through
graft polymerization of acrylic acid (AA) monomers onto okara backbones
in the presence of N,N′-methylenebisacrylamide (MBA) as crosslinker. The
initiator ammonium persulfate (APS) was heated to generate free radicals
on okara particles, and the obtained okara macroradicals initiate the graft po-
lymerization. To achieve this, 7.5 wt% okara suspension was prepared by
adding water to fresh okara. The suspension was homogenized, purged by
N2 gas for 30 min, before increasing the temperature to 70°C under N2
flow. A fixed amount of APS was added to the okara suspension while main-
taining the temperature at 70°C under N2 flow for 30 min. Subsequently,
predetermined amounts of partially neutralized AA and MBA in water were
added. The resulting product after 5 h was oven-dried and milled to powder.
2.2. Plant materials and growth conditions
Choy sum [Brassica rapa L. subsp. chinensis (L.) Hanelt var. parachinensis
(L. H. Bailey) Hanelt] (CS) and pak choi (Brassica rapa subsp. chinensis)
(PC), two commonly consumed Asian leafy vegetables belonging to the
family Brassicaceae, were selected for the field-testing of okara-derived
hydrogel (OP). As a general practice in commercial farms, both cultivars
are grown in two stages, first (Stage 1), from seeds to seedlings prior to
transplanting, and second (Stage 2), from transplanted seedlings to adult
plants before the vegetables are harvested for consumption. In this study,
the vegetables were grown in the above two stages, in sheltered greenhouse
[19.5 m (L) × 5.5 m (W) × 2.5 m (H)] at Sembawang, Singapore, for better
monitoring of irrigation.
Briefly, seeds of vegetables were first sown in germination trays (100-cell;
34 cm × 28 cm × 4 cm) containing growing substrate (Jiffy® Substrates;
Toul, France) till 14 days after sowing (D14; Stage 1). One seedling was
grown in each cell and seedlings were irrigated thrice daily through a
J. Zhu et al.
sprinkler system. At Stage 2, 14-day-old seedlings were transplanted to the
ground in regular-spaced (about 10 cm apart) pre-conditioned raised beds
(25 cm in height) till 30 days after sowing (D30) before the plants were har-
vested for end-point analyses (Section 2.3). These raised beds were prepared
a week before transplanting by mixing the topsoil with chicken manure
(1.5 kg m−2). The soil prepared for vegetable cultivation generally contained
5.1 % organic matter, had an EC value of 0.48 mS cm−1 and with a soil pH of
about 7.35. A total of 25 plants were grown in each plot (0.5 m by 0.5 m) and
duplicate plots were prepared per treatment.
To test the efficacy of OP on plant growth, various concentrations
(0–0.4 %, w/v) of OP granules were added to the soil during transplanting.
Grooves (<10 cm deep) between two lines of transplanted 14-day-old seed-
lings were prepared for OP to be embedded before irrigation was performed
through pre-programmed sprinkler system. Similar plots (without plant
growth) were prepared for soil collection (Section 2.5). Three watering
regimes [twice a week, once a day (7 times a week) and thrice a day
(21 times a week, which is the current standard practice), at 1.5 L per
min per sprinkler, 3 min per irrigation] were programmed to determine
the effect of watering frequency in the presence or absence of the hydrogel
on the growth of the vegetables. Fertilizers (1.5 g of NPK 15:15:15/bed)
were applied on D16 in pellet form, while another on D23 through
drenching. All plants were maintained under greenhouse conditions with
temperature ranging from 34 ± 10 °C/25 ± 3 °C (day/night) and average
relative humidity of 68/87 % (day/night).
2.3. End-point growth assessment of plants
Growth assessment was focused on above-ground biomass. All plants
(450 plants per cultivar from 18 plots) were harvested for end-point analy-
ses on D30. Total shoot fresh weight (FW) per plot was determined. In ad-
dition, 15 plants per plot were randomly selected and destructively
sampled for biomass and leaf area analyses. Shoot FW of the individual
plants were recorded (Mettler Toledo ML303 Precision Balance; Greifensee,
Switzerland) at harvest and samples were dried in the oven at 60 °C to ob-
tain the shoot dry weight (DW) of individual plants. Shoot weights of a total
of 30 plants were recorded per treatment. Relative growth rates (RGR) were
then determined according to Tan et al. (2020) using the following
equation:
lnW2 − lnW1
RGRðFW or DWÞ ¼
t2 −t1
where:
W1 is the initial shoot FW or DW at D14;
W2 is the final shoot FW or DW at D30;
t2-t1 representing the growth period between harvesting (D30) and
transplanting stage (D14).
To determine the “leafiness” of the vegetables, the total leaf area per plant
was recorded according to Tan et al. (2020). The leaf laminae of each plant
were cut and spread out before photographs were taken, followed by area
determination using Image J (version 1.51; National Institute of Health; Be-
thesda, MD, USA). Leaf areas from 16 plants per treatment were determined.
2.4. Irrigation water use efficiency
Irrigation water use efficiency (IrrWUE) was calculated according to
Abuarab et al. (2013) and Tan et al. (2021) with minor modifications.
Briefly, the above-ground FW of plants at harvest (D30) and the total
volume of water used for irrigation were recorded. IrrWUE was then
calculated based on the following equation:
Ws
IrrWUE ¼
Vi
3
Science of the Total Environment 851 (2022) 158141
where
IrrWUE is the g increase in shoot FW per kg of water irrigated;
Ws is the shoot FW at D30 (in g);
Vi is the volume of water used to irrigate the plant (in kg) from D14 to
D30.
2.5. Nutrient analysis of soil extract
Soil from control plots (1 m by 1 m; without any plant cultivation) under
the various watering frequencies (2, 7 or 21 times per week) and different
OP concentrations (0, 0.2 or 0.4 %; w/v; Section 2.2) were sampled using
a tubular soil sampler on the same day (i.e., D30) when the vegetables
were harvested for analyses. At least three areas within each plot were sam-
pled. To determine the nutrient concentrations in the soil, nutrient extrac-
tion was performed according to Sorn-Srivichai et al. (1988) with
modifications. Briefly, 20 mL of compactly-packed soil was prepared
from each of the soil sample collected from the field and transferred to a
200-mL conical flask. To this, 50 mL of water was added and mixed thor-
oughly to obtain a suspension before they were left in an orbital shaker in-
cubator (IKA Works Inc., Staufen, Germany) at 100 rpm, room temperature
for 24 h. Each soil sample was then centrifuged (Eppendorf AG, Hamburg,
Germany) at 3000 rpm, at 4°C for 20 min and filtered through Whatman®
filter paper to obtain the filtrate. The filtrates were analysed for potassium
through quantitative metal analysis (Avio 500 Inductively Coupled Plasma-
Optical Emission Spectrometer (ICP-OES); PerkinElmer Inc., Waltham, MA,
USA), as well as nitrates and phosphates using anion chromatography
(Metrohm Ion Chromatography System; 820 IC Separation Centre;
Metrohm AG, Herisau, Switzerland). Results were expressed as ppm present
in the soil extract.
2.6. Statistical modelling and analysis
All statistical analyses were performed using RStudio Version 1.3.959
(Team, 2020). A two-way ANOVA showing significance of the main effects
[i.e., concentration of OP (G) and watering frequency (W)] and their inter-
actions (G*W) for the variables examined were performed. In situations
where significant interactions between the main effects were observed,
separate comparisons of one-way means were conducted. In addition,
post-hoc test (Tukey's HSD) with significance level set at p <0.05 were
performed following two-way ANOVA analyses, where pairwise compari-
sons were interpreted for the significant factors from the two-way
ANOVA outcomes. When significant interactions between the main effects
were observed, only post-hoc tests for the significant interacting factors
(i.e., G*W) were considered. Where appropriate, a one-way analysis of
variance (ANOVA) followed by post-hoc Tukey pairwise analysis with sig-
nificance level set at p <0.05 was applied to determine if the various treat-
ments affected growth and IrrWUE of each cultivar. In addition, partial
least squares (PLS) biplot was prepared using SIMCA 14.0 (Umetrics,
Sweden).
Statistical modelling was performed to determine the best models for
yield and water use efficiency predictions. The modelling approach was
adopted according Teoh et al. (2019) with modifications. All independent
variables were first tested for multicollinearity by fitting the variables to a lin-
ear model and the variance inflation factors (VIF) determined. For yield pre-
diction, all independent variables on shoot fresh weights (FW) were refitted
to a generalized least squares (gls) model for data visualization and normality
assumption assessment. Comparison of the various variance structure was
conducted and selected based on the lowest Akaike information criterion
(AIC) value. Where normality assumptions were violated, log transformations
of data were performed and compared against the original data.
In view of possible non-independence between plots, the explanatory
variables fitted to a gls model were compared with linear mixed effects
(lme) models, where the fixed and random effects were taken into consid-
eration. For lme models, random intercept model as well as random inter-
cept and slope model were included. AIC values and likelihood ratio were
J. Zhu et al. Science of the Total Environment 851 (2022) 158141

considered for model selection. A top-down approach was adopted (Zuur
et al., 2009), starting with a maximal lme model to select for the one with
the lower AIC value. Alternatively, the models were weighted using the
MuMIn package and the model with the lowest AICc was selected and
refitted using the restricted maximum likelihood (REML) method for
parameter estimation. For model validation, diagnostic plots were created
for the selected model to inspect for any violations in normality and homo-
scedasticity assumptions.
The model selection process for yield prediction was repeated for more
precise estimation of the water use efficiency by replacing the response
variable with IrrWUE. Model validation of the final model was performed
to inspect for violations in the assumptions of normality and homoscedas-
ticity, and the model selection process was repeated if such violations
were identified. Predict plots were then presented based on the selected
best models for yield and water use efficiency.
concentration to 0.4 % (w/v), however, did not increase the average
shoot FW (16 ± 1 g) further. When watering frequency was increased to
21 times per week, the average shoot FW obtained for CS grown without
OP, 0.2 % (w/v) and 0.4 % (w/v) OP were 26 ± 2 g, 24 ± 2 g and 23 ±
1 g, respectively.
For pak choi (PC), addition of 0.2 % (w/v) OP resulted in >20 %
increase in the average shoot FW regardless of watering frequency as
compared to without OP (Fig. S1A). Such an increment in shoot FW was
more pronounced, with plants growing with this concentration of OP
showing >35 % increase as compared to plants that were grown without
OP when the plants were water 7 times per week, and better performance
could be observed for CS over PC. Nevertheless, the application of 0.2 %
(w/v) OP led to dual benefits of significantly increased plant yield
combined with a three-fold reduction in the water application frequency
in both vegetable types under study.

3. Results 3.2. Shoot dry weight (DW)

3.1. Shoot fresh weight (FW)
When choy sum (CS) was irrigated twice a week, the average shoot FW
obtained was 17–20 g (Fig. 1A). At watering frequency of 7 times per week,
higher average shoot FW of 31 ± 2 g per plant was obtained for plants
grown with 0.2% (w/v) OP. This was >60 % increase compared to the
plants without OP under the same watering regime. Increasing OP
At a watering frequency of twice a week, the average shoot DW was
0.8 ± 0.1 g for CS grown without OP. The DW increased to 1.2 ± 0.1 g
and 1.0 ± 0.1 g when plants were grown with 0.2 % and 0.4 % (w/v)
OP, respectively (Fig. 1B). Highest average shoot DW (1.6 ± 0.1 g) was
obtained for plants grown with 0.2 % (w/v) OP and watered 7 times a
week (Fig. 1B), which was >75 % increase as compared to the plants with-
out OP and degree of increment was higher than the shoot FW obtained.

Fig. 1. Effect of Ok-PAA (OP) and watering frequency on the growth and WUE of CS. Plants were grown under field conditions (n = 25 per plot) in the presence of 0–0.4 %
(w/v) OP and were watered under various watering frequencies (2, 7 and 21 times/week) before the plants were harvested (at D30) for end-point analyses. (A) Shoot fresh
weight (FW; n = 30), (B) shoot dry weight (DW; n = 16), (C) total leaf area per plant (n = 16), and (D) irrigation WUE (IrrWUE) were determined. Where possible, two-way
ANOVA were performed to determine significance in response to treatments (p <0.05; refer to Table 2).

4
J. Zhu et al.
For PC, >34 % and 27 % increase in average shoot DW were observed
for plants grown with OP at a watering frequency of twice and 7 times
per week, respectively, as compared to those without OP (Fig. S1B). At a
watering frequency of 21 times per week, the increase in shoot DW was
20 % and 49 %, respectively, for PC grown with 0.2 % (w/v) and 0.4 %
(w/v) OP compared to plants grown without. Hence, application of OP
generally increased both the shoot FW and shoot DW of CS and PC.
3.3. Leaf area
When CS was irrigated twice a week, the total leaf area (per plant basis)
for plants without OP was 331 ± 21 cm2, compared to 365 ± 27 cm2 and
338 ± 27 cm2 for plants with 0.2 % (w/v) and 0.4 % (w/v) OP, respectively
(Fig. 1C). At a watering frequency of 7 times per week, supplementation of
OP at 0.2 % (w/v) increased the total leaf area to 434 ± 28 cm2 without
significantly changing the total number of leaves per plant, a 41 % incre-
ment in leaf area as compared to plants grown without OP.
In terms of “leafiness” for PC, irrigating twice per week produced
260–330 cm2 of leaf area (per plant basis) and plants grown with 0.2 %
(w/v) OP produced smaller leaves (12–21 % decrease) compared to those
without OP (Fig. S1C). On the contrary, at a watering frequency of
7 times per week, a total leaf area of 284 ± 20 cm2 per plant was obtained
with 0.2 % (w/v) OP, which was >21 % increase compared to plants grown
without OP. Under the well-watered condition of 21 times of irrigation per
week, plants with larger leaf area per plant without significantly changing
the total number of leaves per plant were obtained in the presence or
absence of OP, and the total leaf area per plant were in the range of
314–351 cm2.
3.4. Irrigation water use efficiency (IrrWUE)
To assess if watering frequency and addition of OP could improve irriga-
tion water use efficiency (IrrWUE), the crop yield per kilogram of water
irrigated on CS was determined. Higher IrrWUE was observed with lower
watering frequency regardless of OP supplementation, with 102–114 g
FW, 41–82 g FW and 20–22 g FW CS obtained per kilogram of water
irrigated when the plants were watered 2, 7 or 21 times per week, respec-
tively (Fig. 1D). However, the degree of increase in the IrrWUE with respect
to watering frequency was different in the presence of OP.
With two irrigations a week, the IrrWUE for plants grew without OP was
107 ± 7 g FW/kg water irrigated, compared to 6.4 % increase in the
IrrWUE (114 ± 8 g FW/kg water irrigated) for plants that were grown
with 0.2 % (w/v) OP (Fig. 1D). At a watering frequency of 7 times per
week, the IrrWUE for plants without OP was 50 ± 2 g FW/kg water
irrigated, while 82 ± 6 g FW/kg water irrigated was obtained when plants
were grown with 0.2 % (w/v) OP, which was a 63 % increase with respect
to the plants grown without. IrrWUE did not vary much under well-watered
condition of 21 times irrigation per week for plants that were grown with or
without OP.
In addition, decreasing the watering frequency from 21 to 7 times per
week doubled the IrrWUE for plants grown without OP, compared to a
four-fold increase when plants were grown with 0.2 % (w/v) OP under
the same watering frequency. Further increase in the OP concentration
Table 1
Effect of OP concentration and watering frequency on the relative growth rates (RGR) of CS. Results are expressed on a fresh weight [RGR(FW)] and dry weight [RGR(DW)]
basis.
OP (%) RGR(FW) (day−1)a
Watering frequency (No. of times per week)
2 7 21
0 0.264 ± 0.006 0.270 ± 0.003 0.286 ± 0.005
0.2 0.269 ± 0.005 0.298 ± 0.005 0.280 ± 0.005
0.4 0.259 ± 0.006 0.253 ± 0.005 0.281 ± 0.003
a
A two-factor ANOVA is performed (refer to Table 2); n = 30 and 16 for RGR(FW) and RGR(DW), respectively.
5
Science of the Total Environment 851 (2022) 158141
did not result in a concomitant increase in the IrrWUE. Comparing plants
that were irrigated 21 times per week with those watered twice a week, a
4.8-fold increase in IrrWUE was observed for those grown in the absence
of OP, compared to 5.6- and 5.1-fold increase in the IrrWUE for those that
were grown with 0.2 % (w/v) and 0.4 % (w/v) OP, respectively (Fig. 1D).
A more pronounced effect of 35 % increment (i.e., 62 g FW/kg water
irrigated) in IrrWUE was observed when PC were irrigated at 7 times per
week with 0.2 % OP, compared to those grown without (i.e., 46 g FW/kg
water irrigated; Fig. S1D). In both vegetable types, the steepest trendline
obtained for plants grown with 0.2 % (w/v) OP in response to decreasing
watering frequency (Figs. 1D and S1D) further indicated that watering
frequency could be reduced and that better irrigation WUE could be
achieved with the addition of 0.2 % (w/v) OP.
3.5. Relative growth rates (RGR)
Calculated either on a fresh or dry weight basis, CS grown with 0.2 %
(w/v) OP showing the highest RGR, especially when the watering fre-
quency was reduced to 2 or 7 times per week (Table 1). When CS grown
with 0.2 % (w/v) OP were watered 7 times per week, an RGR(FW) of
0.30 ± 0.01 day−1 was obtained, which was a 10 % increase as compared
to the plants grown without OP. Under well-watered conditions of 21 times
of irrigation per week, the RGR(FW) did not differ much with or without OP.
For PC, irrigating twice or 7 times per week resulted in 4 % and >8 %
increase in the RGR(FW), respectively, for plants that were grown with
0.2 % (w/v) OP, as compared to those without (Table S3). Further increase
in the watering frequency to 21 times per week resulted in a 5 % increase in
RGR(FW) for PC grown with 0.4 % (w/v) OP compared to those without.
In terms of RGR(DW), about 5–10 % increase was observed for CS
grown with 0.2 % (w/v) or 0.4 % (w/v) OP as compared to those without
when the plants were irrigated twice a week (Table 1). Increasing the
watering frequency to 7 times per week led to a 15 % increase in the RGR
(DW) (i.e., 0.26 ± 0.01 day−1) for CS grown with 0.2 % (w/v) OP as
compared to those without OP.
Similarly for PC, the RGR(DW) was generally higher when plants were
grown with 0.2 % (w/v) OP regardless of watering frequency. For example,
under a watering frequency of twice a week, a 9 % increase in the RGR(DW)
was obtained for plants grown with 0.2 % (w/v) OP compared to those
grown without OP (Table S3). Addition of 0.2 % (w/v) OP in the growing
medium also resulted in >6 % increase in the RGR(DW) for plants that
were irrigated 7 or 21 times per week (Table S3).
3.6. Two-way ANOVA
Statistical inference showed that OP concentration (G) and watering fre-
quency (W) are the key factors that could singly affect shoot FW (p <0.001)
significantly, and showed significant interaction (p <0.001) between OP
concentration and watering frequency (G*W; Table 2) for both CS and
PC. This implies that the effect of OP concentration on shoot FW is also
dependent on the watering frequency. One-way means further showed
that the positive effect of OP at 0.2% (w/v) on the shoot FW as compared
to those grown without OP (Fig. S2A) was significant, while decreasing
the watering frequency from 21 to 7 times a week did not significantly
RGR(DW) (day−1)a
2 7 21
0.218 ± 0.004 0.224 ± 0.003 0.243 ± 0.007
0.238 ± 0.007 0.257 ± 0.005 0.243 ± 0.009
0.229 ± 0.007 0.224 ± 0.006 0.239 ± 0.004
 J. Zhu et al.
Table 2
Two-factor ANOVA showing the significance of the two main effects (OP concentration, watering frequency) and their interactions on the growth of CS or PC. Growth
parameters such as shoot fresh weight (Shoot FW), shoot dry weight (Shoot DW), relative growth rates on a FW [(RGR(FW)] and DW [(RGR(DW)] basis, leaf area per plant
(Leaf Area), and irrigation water use efficiency (Irrigation WUE) were taken into consideration.
Plant Cultivar Factors/Interactions Shoot FW Shoot DW
(g) (g)
CS OP concentration (G) ⁎⁎⁎ ⁎⁎⁎
Watering frequency (W) ⁎⁎⁎ ⁎ ⁎⁎⁎
G*W ⁎⁎⁎ NS
PC OP concentration (G) ⁎⁎⁎ ⁎⁎⁎
Watering frequency (W) ⁎⁎⁎ ⁎⁎
G*W ⁎⁎⁎ NS
NS: not significant (p-value ≥0.05).
⁎⁎⁎ p-value <0.001.
⁎⁎ p-value <0.01.
⁎ p-value <0.05.
affect shoot FW (Fig. S2B). In addition, post-hoc Tukey's HSD tests focusing
on the significant interacting factors (G*W) showed that most of the
pairwise comparisons for CS grown at 0.2% (w/v) OP with a watering
frequency of 7 times per week were significantly different (Table S1).
Interestingly, post-hoc Tukey's HSD tests focusing on the significant
interacting factors (G*W) on the growth of PC showed that plants grown
at 0.4% (w/v) OP with a watering frequency of 21 times per week produced
significantly higher shoot FW compared to 0.2 % (w/v) OP at the same
watering frequency, and without hydrogel supplementation but at a
lower watering frequency of 2 or 7 times per week (Table S4).
Similarly, shoot DW for both vegetables was significantly affected by OP
concentration (p <0.001) and watering frequency (p <0.05 for CS and
p <0.01 for PC). However, no significant interaction between these two
main effects (G*W, Table 2) was observed, indicating that OP concentration
and watering frequency can singly affect shoot DW. Post-hoc Tukey's HSD
tests focusing on these significant main effects further showed that OP at
0.2% (w/v) can significantly affect shoot DW as compared to those grown
without or at 0.4% (w/v) OP for both CS (Table S2) and PC (Table S5).
Two-way ANOVA analyses showed significant effect of OP concentra-
tion (p <0.05) for CS and watering frequency (p <0.01 for CS and
p <0.001 for PC) for both vegetables on the leaf area. However, no signifi-
cant interaction between these two factors (G*W) was observed for both
(Table 2). Post-hoc Tukey's HSD tests showed that leaf area obtained from
CS grown with 0.2 % and 0.4 % (w/v) OP was significantly different
(Table S2). Similar post-hoc tests on PC indicated that watering frequency
of 7 and 21 times can significantly affect the leaf area production in CS
(Table S2) and PC (Table S5). These indicate that the concentration of OP
or water frequency alone can significantly affect the leaf area of CS while
watering frequency but not OP concentration significantly affected the
leaf area of PC.
In terms of water use efficiency, both OP concentration (p <0.001 for CS
and p <0.05 for PC), and watering frequency (p <0.001) significantly
affected IrrWUE (Table 2) for both CS and PC. Significant interaction
between these two factors was observed for CS (G*W; p <0.01) but not
PC, implying that the effect of OP supplementation on IrrWUE was also
dependent on the frequency of watering for CS (Table 2; Fig. S2). One-
way means and post-hoc Tukey's HSD tests on the significant interacting
factors further showed significant improvement in the IrrWUE when CS
were grown with 0.2 % (w/v) OP (Fig. S2A and Table S1) regardless of
watering frequency. Significant decrease in IrrWUE with increasing
watering frequency from twice a week to 7 times a week, to 21 times per
week of irrigation regime were observed for CS (Fig. S2B). As for PC,
these findings imply that although not as pronounced as CS, the watering
frequency can be reduced from 21 times per week, and that better IrrWUE
can be achieved with the addition of 0.2% OP during the cultivation of PC
(Fig. S2B and Table S5).
Growth rates were significantly affected by OP concentration or
watering frequency (p <0.001–0.05) for CS and significant interaction
between OP concentration and watering frequency (G*W) on RGR was
6
Science of the Total Environment 851 (2022) 158141
RGR(FW) (day−1) RGR(DW) Leaf Area Irrigation WUE
(day−1) (cm2) (g FW kg−1 water irrigated)
⁎⁎⁎ ⁎⁎⁎ ⁎ ⁎⁎⁎
⁎ ⁎⁎ ⁎⁎⁎
⁎⁎⁎ ⁎ NS ⁎⁎
NS ⁎⁎⁎ NS ⁎
NS ⁎ ⁎⁎⁎ ⁎⁎⁎
⁎ NS NS NS
also observed (Table 2). One-way means and post-hoc Tukey's HSD tests
on the significant interacting factors indicated significant positive effect
of OP at 0.2% (w/v) on RGR(DW) as compared to without OP supplemen-
tation (Fig. S2A and Table S1), while decreasing the watering frequency
from 21 to 7 times a week did not significantly affect the RGR(FW) and
RGR(DW) (Fig. S2B).
For PC, OP concentration or watering frequency alone did not signifi-
cantly affect the RGR(FW), but significant (p <0.05) interaction between
OP concentration and watering frequency (Table 2) was observed, indicat-
ing that combination of OP concentration and watering frequency signifi-
cantly affected RGR(FW).
3.7. Nutrient analysis of soil extract
To determine if OP can help in retaining the nutrients in the soil,
analyses of nitrates (N), phosphates (P) and potassium (K) were performed
on soil samples collected from control plots without any plant growth,
prepared together at the beginning of the experiment. Analyses of the soil
extract at the end of the experiment generally showed higher concentra-
tions of nutrients being present in the soil when OP was incorporated
(Fig. 2). Except for N (Fig. 2A), increasing concentrations of P (Fig. 2B)
and K (Fig. 2C) were detected in the presence of OP in a dose-dependent
manner regardless of watering frequency. Less than 30 ppm P and
50 ppm K could be detected in the soil extract without OP, compared to
>50 ppm P and 70 ppm K with the addition of 0.2 % (w/v) OP. Doubling
the OP concentration resulted in significantly higher concentrations of P
(>80 ppm) and K (>200 ppm) detected in the soil extract when the plots
were irrigated 7 or 21 times per week, as compared to without OP under
the same watering frequency (Fig. 2B, C). These results suggested that
more P and K could be retained in the soil following OP amendment.
3.8. PLS biplot and model prediction
PLS biplot integrating the partial least squares (PLS) plot and loading
plot was further used to assess the growth and water use performance of
two vegetable types in response to OP at a watering frequency of 7 times
per week (Fig. 3). Principal Components 1 (PC1) and 2 (PC2) accounted
for 88.7 % and 9.4 % of the variation, respectively, amounting to an eigen-
value of >98 %. These results indicated that incorporation of OP at 0.2 %
(w/v) improved yield and water use efficiency compared to without OP,
as reflected by shoot FW, RGR, leaf area and IrrWUE.
To determine the best models for yield and water use efficiency predic-
tions, all independent variables were first fitted to a linear model and VIF
values indicated no problem of multicollinearity among the variables. Gls
models fitted with different variances per stratum (varIdent) generally
improved the model, with the variance structure taking OP concentration
and watering frequency into consideration giving the lowest AIC values
for both yield and water use efficiency models (Table S6). A relatively
straight line could be observed from the Q-Q plots for yield and water use
J. Zhu et al.
Fig. 2. Effect of OP on nutrient retention in the soil. Soil samples (n = 3) with or
without OP supplementation under various watering frequency (i.e., 2, 7 or 21
times of water irrigation per week) collected from the control plots (without plant
cultivation) were analysed for their (A) nitrates, (B) phosphates and (C) potassium
(K) content at the end of the experiment. Nutrient contents were compared within
each watering frequency. Asterisk (*) indicates significance (p <0.05; one-way
ANOVA) with respect to control (i.e., without OP supplementation).
efficiency models following log transformation of data, indicating that the
residuals in these models were relatively normal (Fig. S3). Based on the
lowest AIC values, a gls model fitted with a varIdent variance structure
was selected as the best model for yield prediction while a lme model
with one candidate random effect (i.e., plot) was selected for water use
prediction (Table S7). Graphical validation of these selected models further
showed that there were no issues on heteroscedasticity and non-normality
for the selected models (Fig. S4).
Without OP, highest yields for both CS and PC were predicted to be
when the watering frequency was set at 21 times per week (i.e., thrice
per day) (Fig. 4A). Whereas, with the addition of 0.2 % (w/v) OP, the
highest predicted yields for both vegetable types could be achieved when
7
Science of the Total Environment 851 (2022) 158141
Fig. 3. PLS biplot of variables distribution obtained from assessing the growth and
water use performance of two vegetable types in response to OP supplementation at
a watering frequency of 7 times per week. Leaf area (Area), relative growth rate
calculated on a fresh weight basis (FWRGR), irrigation water use efficiency
(IrrWUE) and shoot fresh weight (ShootFW) of CS (closed circles) and PC (opened
circles) grown in soil supplemented with various concentrations of OP (%; w/v)
at 0: pink; 0.2 green; 0.4: blue, were analysed.
the watering frequency was reduced to 7 times per week (Fig. 4A). In
brief, the highest yields for both vegetables were predicted to be in the
presence of 0.2 % (w/v) OP and irrigated 7 times a week (i.e., once daily)
among all the combinations of gel concentrations and water frequencies
evaluated.
In terms of water use efficiency, better water usage was observed for
both CS and PC when the plants were irrigated twice a week, regardless
of OP supplementation (Fig. 4B). However, best water use efficiency was
predicted to be when both vegetable types were grown with 0.2 % (w/v)
OP. The same concentration of OP was also predicted to be the best for
both vegetable types if moderate irrigation of 7 times per week was applied
to the plants during cultivation (Fig. 4B).
4. Discussion
In this study, we demonstrated the suitability of applying okara-based
hydrogel (OP) on the growth of two different green leafy vegetables
under field conditions. We have previously shown growth enhancement
of CS seedlings under water-limited conditions when the potting mix was
supplemented with a prototype of okara-derived superabsorbent hydrogel
(Gel-1) developed earlier (Zhu et al., 2020a; Zhu et al., 2020b). We further
demonstrated an increase in yield (by 88 % and 113 %) with improved
irrigation WUE when the vegetables were grown in substrate supplemented
with Gel-1 and maintained individually in pots under indoor conditions
(Tan et al., 2021), where the growth conditions of the plants were more
constant and could be better managed.
Under field conditions, the vegetables were grown on the ground in
raised beds, with growth conditions being less predictable and fluctuate
with the environmental conditions. As shown in Fig. 1, Tables 1 and 2,
either supplementation with OP (0.2–0.4 %; w/v) or changing watering
frequency significantly affected all the growth parameters assessed (shoot
FW, shoot DW, RGR, leaf area, irrigation WUE) under various watering
regimes for CS under field conditions. The findings are similar for PC,
except for the RGR(FW) when considering the main factors alone, or leaf
area, as far as OP concentration is concerned (Fig. S1, Tables 2 and S3).
Significant interactions between OP concentration and watering frequency
(i.e., G*W) observed for shoot FW and not shoot DW seem to imply these
J. Zhu et al.
Fig. 4. Fitted values of the best supported models by AIC. Note the predicted highest
yield (A) and best water use (B) obtained for CS (circled in blue) and PC (circled in
green) which is a combination of OP at 0.2 % (w/v) and watering frequency at 7
times per week. This probably explains why the interaction term is significant
(Table 2).
interacting factors affecting the water content of the plants but not so much
on the shoot biomass per se. Nevertheless, significant interactions between
the main effects were generally observed for the growth and water use
efficiency of CS but less so for PC (Tables 2, S1, S2, S4 and S5). Together
with PLS biplot analysis and the prediction based on selected models for
yield and water use, vegetables grown in soil with 0.2 % (w/v) OP and
with a watering frequency of 7 times per week showed the best growth
performance and water use efficiency (Figs. 1, 3, 4 and S1). Overall, yield
increase could still be observed under more challenging and less
predictable field conditions, with shoot fresh weight and leaf area showing
>60 % and 40 % increase, respectively for CS, and >35 % and 20 %
increase, respectively for PC.
8
Science of the Total Environment 851 (2022) 158141
Water-retention supplement/additives potentially increase water
availability in the substrate, thereby resulting in better plant growth and
survivability without the need to provide large volumes of extra growing
media, with hydrogels being the most widely-deployed additives for
agricultural, horticultural or forestry applications (Agaba et al., 2010;
Arbona et al., 2005; Martin del Campo et al., 2019; Paradelo et al., 2019;
Satriani et al., 2018; Shi et al., 2010). Arbona et al. (2005) reported
substrate amendment with 0.4 % hydrogel (Stockosorb Agro) increased
the survivability of citrus seedlings and alleviated the drought stress effects
on citrus plants through reduction in the degree of leaf abscission, improve-
ment in photosynthesis rate, stomatal conductance and root growth.
Similar findings were reported by Shi et al. (2010) on Populus popularis.
Agaba et al. (2010) tested on several tree seedlings, including Eucalyptus
grandis, Pinus caribaea, Araucaria cunninghamii, and Terminalia superba,
and showed that soil amendment with Luquasorb could significantly
increase the plant available water and prolong the survivability of the
trees compared to the controls without hydrogel amendment in response
to drought stress. A recent study by Farsaraei et al. (2020) working on
sweet basil (Ocimum basilicum) reported 4 % and 19 % shoot weight
increase with the application of Ackoasorb and Stockosorb, respectively,
in contrast to plants that were grown without hydrogel supplementation.
All in all, the hydrogels evaluated in these studies are petroleum-based
and none are on green leafy vegetables.
In addition to synthetic petroleum-based SAP, there has been an
increasing trend in the upcycling of waste materials for the production of
natural superabsorbent hydrogels. For examples, a cellulose-g-poly (acrylic
acid-co-acrylamide) superabsorbent composite based on flax yarn waste
(Wu et al., 2012) and mulberry branch-g-poly(acrylic acid-co-acrylamide)
hydrogel (Zhang et al., 2014) had been reported. Zhang et al. (2021) had
utilized the biowaste white cabbage to synthesize their superabsorbent
polymer gel. Nanocellulose extracted from agricultural wastes (such as
rice straw, wheat straw, corn straw) for the synthesis of cellulose-based
superabsorbent hydrogels to be used as soil supplements had also been
reviewed by Li and Chen (2020). While Songsrirote et al. (2017) reported
the synthesis of poly(vinyl alcohol) (PVA)/Glutaraldehyde (GA)/okara
hydrogel through microwave-assisted irradiation, however information
on its use as a potential soil supplement is lacking. To the best of our knowl-
edge, this is the first comprehensive report on the efficacy of an okara-based
hydrogel on the growth of vegetable crops under field conditions besides
hydrogel characterization. With improved water absorbency and its ability
to enhance water holding and retention properties of soil (Zhu et al., 2022)
being similar to petroleum-based hydrogels, this biomass-derived hydrogel
(OP) has been shown herein to be able to enhance vegetable yield increase
while decreasing water usage, and has the added advantage of using renew-
able resources and upcycling waste.
Water is one of the most critical environmental components and the
demand for plant cultivation is challenging, with 70 % of our freshwater
resources used for agriculture (Bourzac, 2013). This demand for plants
with larger leaf area, such as the green leafy vegetables are particularly
high, as they need to cope with the substantial water loss through the
stomata as a result of high transpiration rates, especially in tropical regions
where ambient temperatures tend to be higher. Frequent irrigation is
required to compensate for the poor water retention ability of the soil in
order for the plants to stay hydrated, often with excessive watering leading
to unnecessary water wastage and nutrient leaching (Tan et al., 2021).
Higher impact could be felt in urban and peri-urban farming where water
is required to be transported from long distances (Bandla et al., 2019). As
demonstrated here, in addition to yield increase, watering frequency
could be reduced and better irrigation WUE could be achieved with the
addition of 0.2% (w/v) OP in the soil (Figs. 1D and S1D). El-Asmar et al.
(2017) showed incorporation of 0.2–0.4 % of hydrogel (STOCKOSORB®
660) in soil resulted in 10–13 % increase in water use efficiency in corn.
Our results showed 62 % and 35 % improvement in the irrigation WUE
for CS and PC, respectively, when the vegetables were with 0.2 % (w/v)
OP under a watering frequency of 7 times per week (Figs. 1D and S1D).
For a one-hectare plot, the estimated yield increase would be from 19 t to
J. Zhu et al.
31 t for CS, and from 17 t to 23 t for PC, if the crops were grown with 0.2 %
(w/v) OP. Furthermore, yield increase could be 5 t and 2 t more per hectare
for CS and PC, respectively, in the presence of 0.2 % (w/v) OP with a reduc-
tion in the watering frequency from 21 times to 7 times per week, saving up
to 196,000 L of water per crop cycle per hectare following the incorporation
of OP. The study demonstrates that under field condition substantial
amount of environmental water resources can be protected.
In order to maximize yield, production of vegetables usually involves
intensive application of fertilizers, and together with excessive irrigation,
resulted in nutrient leaching to groundwater, leading to potential environ-
mental pollution and degradation, eutrophication and nutrient losses from
soils (Bijay-Singh and Craswell, 2021; Luijendijk et al., 2020; McDermid
et al., 2021; Wang et al., 2021). In spite of intensive fertilizer application,
world phosphorus (P) deposits, for example, are depleting and P shortage
is predicted worldwide. Assuming absolute shortage of mineral P fertilizer,
>50 % of total P losses in agricultural soils due to erosion by water has been
estimated (Alewell et al., 2020). In the current study, subsequent addition
of OP to the soil did not change the pH substantially (pH = 7.2–7.4; unpub-
lished data). Analyses of the soil collected from control plots (i.e., without
plant growth; Fig. 2) at the end of the experiment generally showed higher
concentrations of nutrients being present in the soil, especially for phos-
phates (P) and potassium (K), in the presence of OP in a dose-dependent
manner regardless of watering frequency (Fig. 2). Application of OP at
higher concentration in the soil, however, did not result in a corresponding
vegetable yield increase (Figs. 1 and S1). Comparing within each watering
frequency, incorporation of 0.4 % (w/v) OP in the soil resulted in a signif-
icant 4- to 6-fold increase in P (Fig. 2B) and 5- to 6-fold increase in K
(Fig. 2C) in comparison to soil without any OP supplementation, indicating
that significantly more nutrients could be retained in the soil following OP
amendment. However, it is not known if such a significant and substantial
retention of P and K in the soil is beneficial for plant growth or the nutrients
retained have already reached the supra-optimal range. Nevertheless, the
ability to retain more nutrients in the soil following OP amendment
would mean more nutrients available in the soil for the plants to tap on,
hence less fertilizer required (either reducing application amount or
frequency) for optimal plant growth, thereby reducing fertilizer usage and
unnecessary wastage and leaching that could lead to further phosphorus
depletion from the soil, groundwater contamination and environmental
pollution (Wu and Liu, 2008).
While intensification of agricultural production usually involves
extensive application of fertilizers to enhance plant yield, it is now
apparent that not >50 % of the fertilizer nitrogen (N) applied is used
by the crops directly (Basu et al., 2022; Bijay-Singh and Craswell,
2021). The remaining of the fertilizers either leached off from soils,
reaching surface and ground water bodies, or add on to the soil organic
N pool where N is mineralized and remained in the soil and contributes
to further leaching for decades. Nitrogen in the form of nitrates is highly
soluble and is a common environmental pollutant to both surface and
ground water bodies, which is further exacerbated with extensive irriga-
tion (Bijay-Singh and Craswell, 2021; McDermid et al., 2021). Such
excessive loss of nutrients not only renders groundwater unsafe for
drinking, it provides nutrient source for coastal and riverine regions
resulted in algal blooms and eutrophication, which eventually leads to
seasonal widespread hypoxia and anoxia that can upset the pristine
marine eco-environments (e.g., coral reefs), impacting on wildlife and
loss of biodiversity (Bijay-Singh and Craswell, 2021; McDermid et al.,
2021). A recent report further shows that eutrophication is an important
driver on the emission of methane, one of the potent greenhouse gases,
from lakes and impoundments that could contribute to the acceleration
of climate change (Beaulieu et al., 2019). Intensive fertilization could
also affect soil quality through accumulation of heavy metals in the
soil eco-system. In addition, prolonged usage of chemical fertilizers
reduces soil organic matter (SOM) content and vital soil minerals and
nutrients, increases soil acidity and soil compaction, which could result
in inadequate aeration, poor drainage, and erosion (Pahalvi et al.,
2021). Together with copious water use for irrigation, this could result
9
Science of the Total Environment 851 (2022) 158141
in enhanced soil salinization, leading to the eventual deterioration
of agricultural land. Through the use of soil supplement such as
hydrogels or OP as reported herein, water holding and retention proper-
ties of the soil could be enhanced, irrigation induced soil water seepage
could be reduced, with increased efficiency in water utilization
(Venkatachalam and Kaliappa, 2021). OP, which is an okara-derived hy-
drogel, not only minimizes watering frequency, but helps in locking-in
nutrients in the soil and prevent excessive leaching (Figs. 1D, S1D and
2), thereby reduces the frequency of using fertilizers. In addition,
using OP has the added advantage of reusing food waste, mitigating
resource losses incurred through waste management, and environmen-
tal impact, as a result of methane and carbon dioxide emissions
from decomposing food, could be reduced (Hall et al., 2009; Tonini
et al., 2018).
As shown in Fig. 4, combining data for the two types of green leafy
vegetables under study, statistical modelling further showed that incorpo-
ration of OP at 0.2 % (w/v) with a two-third reduction in the watering
frequency could have a yield that is comparable to, if not better than
those grown under the well-watered conditions (21 times of irrigation per
week) without any gel supplementation. With the use of advanced statisti-
cal methods and modelling approaches, we have managed to provide some
insightful options for the growers on the choice of OP application and
irrigation regime, as far as a balance between yield and water usage is
concerned. Future work involving more diverse vegetable types and irriga-
tion regimes and/or hydrogel applications would allow more comprehen-
sive data acquisition for more robust modelling and prediction on the use
of the okara-derived hydrogel on the growth performance of crops.
5. Conclusions
As demonstrated here, the use of hydrogel (OP) at 0.2 % (w/v) enhances
the growth of vegetables by 35–60 %, increases the water use efficiency by
30–60 % and minimizes the watering frequency under field conditions,
thus contributing toward water saving. This field study also suggested
that OP amendment significantly retained more soil nutrients and thus
less fertilizers will be needed. Such a waste (okara)-to-resource (soil supple-
ment) approach helps in waste valorization, closing the waste loop of circu-
lar economy for environmentally sustainable agriculture. In short, we have
demonstrated that this plant-based soil supplement can be applied under
field conditions, allowing the plants to stay hydrated for longer periods of
time, which will be especially useful under adverse water-limited and
adverse climatic conditions.
CRediT authorship contribution statement
Wee Kee Tan: Conceptualization, Methodology, Validation, Formal
analysis, Investigation, Data Curation, Writing - Original Draft, Visualiza-
tion, Supervision, Project administration. Jingling Zhu: Conceptualization,
Methodology, Investigation, Resources, Writing - Review & Editing.
Fadhlina Suhaimi: Conceptualization, Methodology, Resources, Writing -
Review & Editing. Jing Ying Lim: Formal analysis, Investigation, Data
Curation, Writing - Review & Editing, Visualization. Zhengyang Gao:
Resources, Writing - Review & Editing. Sanjay Swarup: Conceptualization,
Methodology, Writing - Review & Editing, Supervision, Project administra-
tion, Funding acquisition. Chiang Shiong Loh: Conceptualization,
Methodology, Writing - Review & Editing, Supervision, Project administra-
tion, Funding acquisition. Jun Li: Conceptualization, Methodology,
Resources, Writing - Review & Editing, Supervision, Project administration,
Funding acquisition. Choon Nam Ong: Conceptualization, Methodology,
Writing - Review & Editing, Visualization, Supervision, Project administra-
tion, Funding acquisition.
Data availability
Data will be made available on request.
J. Zhu et al.
Declaration of competing interest
The authors declare that they have no known competing financial inter-
ests or personal relationships that could have appeared to influence the
work reported in this paper.
Acknowledgements
This work was financially supported by the National Research Foundation,
Prime Minister's Office, Singapore, under its Competitive Research
Programme (NRF-CRP 16-2015-04). We would like to thank Prof L. Roman
Carrasco (Department of Biological Sciences, NUS), Drs Zhu Tianming and
Tang Wai Hoh (Data Analytics Consulting Centre, NUS) for their advice on
statistical modelling. Special thanks are given to staff (Sharifah Mohamed,
Ong Pang Hwa, Lee Sook Hiang, Tong Kwong Fatt and Ong Kim Choy) from
the Singapore Food Agency, to Hui Wen, Craig, Asyraf for helping with the
field work, to Zou Li for data presentation for the PCA biplot, and to Kok
Fah Technology Farm Pte. Ltd. (Singapore) for facilitating in the project. We
thank Fortune Food Manufacturing Pte. Ltd. (Singapore) for providing okara
for this research.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2022.158141.
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