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Global Research journal of Natural Science  
& Technology (GRJNST)  
Volume: 04 - Issue 4 (2026), 2111  
ISSN P: 2790-7643 ISSN E: 2790-7651  
www.grjnst.net  
https://doi.org/10.53762/grjnst.04.04.02  
Sustainable Zinc Biofortification of Mung Bean Using Cellulose-Based Slow-  
Release Fertilizer Hydrogel under Normal and Drought Stress Conditions  
Received: 15 April 2026. Accepted: 20 May 2026. Published: 20 June 2026  
Marya Siraj Nabi  
Department of Agricultural Chemistry & Biochemistry,  
The University of Agriculture Peshawar, Khyber Pakhtunkhwa, Pakistan  
Maryasiraj09@aup.edu.pk  
Afia Zia (Corresponding Author)  
Department of Agricultural Chemistry & Biochemistry,  
The University of Agriculture Peshawar, Khyber Pakhtunkhwa, Pakistan  
afia.zia@aup.edu.pk  
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Article ID: 2111  
https://doi.org/10.53762/grjnst.04.04.02  
Copyright © 2026 GRJNST. This article is published under an Open Access model. It is made available to the public under the terms of the Creative  
Commons Attribution 4.0 International (CC BY 4.0) license, which permits unrestricted use and distribution  
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Abstract  
Zinc (Zn) deficiency in edible legumes is a persistent nutritional problem that is  
exacerbated in systems with low micronutrient availability and recurrent  
drought, where conventional fertilizers often show poor synchronization  
between nutrient release and crop demand. This study aimed to evaluate  
cellulose-based slow-release hydrogel fertilizer formulations to improve Zn  
biofortification of mung bean (Vigna radiata L.) under normal irrigation  
(100% field capacity) and drought stress (40% field capacity), and to quantify  
treatment and irrigation effects on Zn accumulation and Zn use-efficiency  
indices. A controlled-environment pot experiment was conducted using a  
factorial arrangement of seven nutrient treatments (control, SRHG5, SRHG10,  
SRFHG5, SRFHG10, NPK-Fe, Zn, and Zn, Fe) across two irrigation regimes  
with three replicates (42 units). Cellulose-derived hydrogels were synthesized  
via cross-linking, loaded with Zn from ZnSO₄·7H₂O (5.6% elemental Zn) at  
an equivalent rate of 25 lb/A (~28 kg/ha; 0.08 g ZnSO₄ per pot),  
characterized for swelling and nutrient-release behavior, and applied near the  
root zone. At physiological maturity, seed and stover Zn concentrations were  
determined after acid digestion using AAS/ICP-OES, and data were analyzed  
by two-way ANOVA. Seed Zn ranged from 29.40-46.57 mg/kg, with  
SRFHG10 achieving the highest values under both 100% (46.57 mg/kg) and  
40% irrigation (38.77 mg/kg); only SRFHG10 met the ≥40 mg/kg  
biofortification target under optimal irrigation, and no treatment reached the  
target under drought. Stover Zn was higher (25.80-59.40 mg/kg), again  
maximized by SRFHG10 (59.40 and 52.00 mg/kg under 100% and 40%  
irrigation). Nutrient treatment significantly affected Zn in seeds (F=9.32,  
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p<0.001) and stover (F=756.26, p<0.001); irrigation had a smaller effect on  
seeds (F=7.16, p=0.01) but a strong effect on stover (F=277.71, p<0.001),  
with a significant treatment×irrigation interaction for stover (p<0.001) but not  
seeds (p=0.57). SRFHG10 also showed the highest Zn use efficiency (38.68  
and 37.04) and agronomic Zn use efficiency (11.00 and 10.82), whereas Zn, Fe  
produced negative agronomic efficiency, indicating poor agronomic response.  
Overall, cellulose-based slow-release fertilizer hydrogel, particularly SRFHG10,  
enhanced Zn accumulation and fertilizer efficiency and partially mitigated  
drought-associated declines, supporting hydrogel-mediated micronutrient  
delivery as a promising strategy for improving mung bean nutritional quality,  
although further optimization is needed for consistent target attainment under  
water limitation.  
Keywords: mung bean, zinc biofortification, hydrogel fertilizer, drought stress,  
zinc use efficiency  
Introduction  
Zinc is an essential micronutrient required for enzyme activation, protein synthesis,  
membrane stability, and reproductive development in crop plants (Alloway, 2008;  
Marschner, 2012). In grain legumes, inadequate Zn supply can reduce productivity and  
limit the nutritional quality of edible seeds, making agronomic biofortification an  
important strategy for improving human nutrition (Cakmak, 2008; White & Broadley,  
2009). Mung bean (Vigna radiata L.) is an important pulse crop because of its short  
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growth cycle, high protein content, and suitability for semi-arid environments; however,  
its productivity and nutrient accumulation are often constrained by drought stress  
(Ranawake et al., 2011; Rashid et al., 2003; Islam et al., 2021).  
Conventional fertilizers may exhibit low nutrient-use efficiency because of leaching,  
fixation, and poor synchronization between nutrient release and crop demand (Fageria,  
2001). In contrast, hydrogel-based delivery systems can improve water retention, reduce  
nutrient loss, and support the gradual release of nutrients under variable moisture  
conditions (Jiang et al., 2020; Peng et al., 2016; Tang et al., 2020). Cellulose-based  
hydrogels are particularly attractive because cellulose is abundant, biodegradable, and  
chemically versatile for agricultural applications (Klemm et al., 2005; Poletto et al.,  
2014; Lohmousavi et al., 2020).  
This study was designed to assess Zn biofortification in mung bean under two irrigation  
regimes using cellulose-based slow-release hydrogel fertilizer formulations. The  
objectives were to compare Zn concentrations in seed and stover among fertilizer  
treatments, determine the effects of nutrient treatments and irrigation using two-way  
analysis of variance, and evaluate Zn use efficiency indices under normal and drought-  
stress conditions.  
Materials and Methods  
Experimental design and site conditions  
The experiment was conducted in a controlled environment (greenhouse or growth  
chamber) on mung bean plants (Vigna radiata L.) under two irrigation regimes: 100%  
irrigation (normal conditions) and 40% irrigation (drought stress conditions). Seven  
nutrient treatments were evaluated in a factorial design with two irrigation levels,  
resulting in 14 treatment combinations. The study followed a completely randomized  
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block design or split-plot arrangement with three replicates per treatment, totaling 42  
experimental units.  
The seven nutrient treatments were as follows: (1) control (no Zn supplementation), (2)  
SRHG5 (slow-release hydrogel with 5.6% Zn), (3) SRHG10 (slow-release hydrogel  
with 10 g/pot), (4) SRFHG5 (slow-release fertilizer hydrogel with 5.6% Zn), (5)  
SRFHG10 (slow-release fertilizer hydrogel optimized), (6) NPK-Fe, Zn  
(conventionally formulated fertilizer with NPK, iron, and zinc), and (7) Zn, Fe (iron-  
zinc mixture without NPK).  
Hydrogel preparation and nutrient loading  
Cellulose-based slow-release hydrogel formulations were synthesized by crosslinking  
carboxymethyl cellulose (CMC) or cellulose derivatives with appropriate polymeric  
agents to create a three-dimensional network structure. The hydrogels were loaded with  
zinc using zinc sulfate (ZnSO4·7H2O) containing 5.6% elemental zinc by mass. Zinc  
was incorporated into the hydrogel matrix at a field-equivalent rate of 25 lb/A  
(approximately 28 kg/ha), corresponding to 0.08 g of ZnSO₄ per pot for the pot-scale  
experimental setup.  
Hydrogel formulations were prepared using chemical or physical cross-linking methods  
to achieve gradual nutrient release kinetics. The hydrogels were characterized for their  
swelling capacity, water-retention properties, and nutrient-release rate under simulated  
soil moisture conditions prior to use in the experiment.  
Plant cultivation and irrigation management  
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Mung bean seeds were sown in pots containing 5-10 kg of air-dried soil (sandy loam or  
loamy texture with moderate fertility). Each pot was planted with two to three seeds at a  
depth of 3-4 cm. Seedlings were thinned to one uniform plant per pot within 7-10 d  
after emergence. Hydrogel-loaded zinc or conventional nutrient treatments were applied  
directly to the soil surface near the root zone at the time of seeding or shortly thereafter  
to allow optimal root-nutrient contact.  
Irrigation was applied based on two irrigation regimes: (1) 100% irrigation-maintained  
soil water at or near field capacity throughout the growing season, and (2) 40%  
irrigation-maintained soil water at 40% of field capacity, simulating drought-stress  
conditions. Irrigation frequency and volume were adjusted based on soil moisture  
measurements using tensiometers or gravimetric sampling. Plants were maintained under  
controlled temperature (25-30 °C Day, 18-22 °C night), ambient relative humidity  
(50-70%), and a 14-h photoperiod with supplemental lighting.  
Sample collection and preparation  
Mung bean plants were harvested at physiological maturity (80-90 days after seeding)  
when pods turned brown and seeds reached maximum dry matter. Harvested plants were  
separated into seeds and stover (stems, leaves, and remaining above-ground biomass).  
Samples were washed with deionized water to remove soil particles, air-dried at room  
temperature for 48-72 hours, and then oven-dried at 70°C to constant weight to  
determine dry biomass.  
Zinc concentration analysis  
Dried seed and stover samples were finely ground to pass through a 200-mesh sieve  
using a Wiley mill or an equivalent grinder. Sub-samples (0.5-1.0 g) of the ground plant  
material were digested using concentrated nitric acid (HNO₃) and/or a mixture of  
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HNO₃ and perchloric acid (HClO₄) in a 2:1 or 3:1 ratio, following standard protocols  
(Chapman & Pratt, 1961; Horwitz, 2010).  
Digestion was conducted in a fume hood at a controlled temperature (120-180 °C)  
until a clear, colorless solution was obtained, after which it was diluted to a fixed volume  
(typically 25 or 50 mL) with deionized water. Zinc concentration in the digested plant  
extracts was determined using atomic absorption spectrophotometry (AAS) or  
inductively coupled plasma optical emission spectrometry (ICP-OES) with appropriate  
standard reference materials. Instrument calibration was verified using certified reference  
standards, and quality control blanks and duplicate samples were included with every  
digestion batch to ensure analytical accuracy.  
Efficiency indices  
Zinc use efficiency (ZUE) was calculated as the ratio of grain yield (kg/ha) to the  
applied Zn (kg/ha). Agronomic zinc use efficiency (AZUE) was computed as the  
difference in yield between the treated and control plots divided by the amount of zinc  
applied. Internal zinc utilization efficiency (IZUE) was calculated as the ratio of grain  
yield to the total zinc content in the seed and stover (mg/kg). The zinc harvest index  
(ZHI) was calculated as the proportion of seed zinc relative to the sum of seed and  
stover zinc, expressed as a percentage, indicating the efficiency of zinc translocation and  
partitioning to the economically important seed fraction.  
Statistical analysis  
Data were analyzed using two-way analysis of variance (ANOVA) to test the main  
effects of nutrient treatment and irrigation regime, as well as their interaction (nutrient  
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× irrigation), on the Zn concentrations in seeds and stover. When the F-test indicated  
significant effects (p ≤ 0.05), mean values were separated using the least significant  
difference (LSD) test or Duncan's multiple range test at a significance level of p ≤ 0.05.  
The coefficient of variation (CV) was calculated as a percentage of the grand mean to  
assess the experimental precision and homogeneity of variance.  
All statistical analyses were performed using SAS statistical software (SAS Institute Inc.,  
Cary, NC, USA) or R statistical programming language (R Foundation for Statistical  
Computing, Vienna, Austria). Normality of residuals was verified using the Shapiro-  
Wilk test, and homogeneity of variance was assessed using Levene's test before  
proceeding with ANOVA. Data were log-transformed when necessary to meet the  
assumptions of parametric statistical tests.  
Results  
Zinc concentration in seeds  
Seed Zn concentration differed among nutrient treatments and irrigation regimes. The  
highest seed Zn concentration was recorded for SRFHG10, reaching 46.57 and 38.77  
mg/kg under 100% and 40% irrigation, respectively, whereas the control treatment  
showed the lowest values of 30.80 and 29.40 mg/kg for the above-mentioned irrigation  
levels. The mean seed Zn concentration across treatments was 36.31 and 33.49 mg/kg  
under normal and drought stress irrigation, respectively (Figure 1).  
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Figure 1. Seed Zn concentration (mg/kg) of mung beans under 100% and 40%  
irrigation across fertilizer treatments. Bars represent treatment means. SRFHG10  
produced the highest seed Zn concentration under both irrigation regimes.  
Two-way ANOVA showed that nutrient treatment significantly affected seed Zn  
concentration (F = 9.32, p < 0.001), whereas irrigation had a smaller but significant  
effect (F = 7.16, p = 0.01). The treatment × irrigation interaction was not significant  
(F = 0.82, p = 0.57), indicating that the relative performance of nutrient treatments for  
seed Zn concentration was broadly consistent across irrigation regimes.  
Zinc concentration in stover  
Stover Zn concentration was higher than seed Zn concentration across all treatments.  
The highest stover Zn concentration was obtained with SRFHG10, with values of  
59.40 mg/kg under 100% irrigation and 52.00 mg/kg under 40% irrigation, while the  
control treatment showed the lowest values of 28.90 and 25.80 mg/kg, respectively.  
Mean stover Zn concentration was 41.24 mg/kg under normal irrigation and 36.10  
mg/kg under drought stress (Figure 2).  
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Figure 2. Stover zinc concentration (mg/kg) of mung bean under 100% and 40%  
irrigation across fertilizer treatments. The bars represent the treatment means.  
SRFHG10 produced the highest stover Zn concentration, and the reduction under  
drought stress remained consistent across treatments.  
Nutrient treatment had a highly significant effect on stover Zn concentration (F =  
756.26, p < 0.001), irrigation also had a highly significant effect (F = 277.71, p <  
0.001), and the treatment × irrigation interaction was also significant (F = 5.73, p <  
0.001). The coefficient of variation was lower for the stover Zn concentration (2.59%)  
than for the seed Zn concentration (9.80%), indicating greater precision in the stover  
measurements.  
Comparative Zn biofortification response  
A comparative analysis of Zn biofortification showed greater Zn accumulation in stover  
than in seeds across all fertilizer treatments. The SRFHG10 treatment produced the  
highest mean Zn concentration in both stover (55.70 mg/kg) and seeds (42.67 mg/kg),  
suggesting superior Zn uptake and translocation efficiency. In contrast, the control  
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treatment yielded the lowest Zn values, confirming that external Zn supplementation  
was necessary to improve Zn enrichment in mung bean tissue.  
Higher Zn retention in stover than in seeds suggests restricted remobilization of Zn to  
reproductive tissues, which is consistent with the known physiological limits to Zn  
transport in edible crops (White & Broadley, 2011). The stronger irrigation effect  
observed in stover than in seeds also suggests that water availability influenced Zn  
uptake and storage in vegetative tissue more strongly than final Zn partitioning to seed  
(Marschner, 2012; White & Broadley, 2009).  
Zinc efficiency indices  
Zinc use efficiency (ZUE) was highest in SRFHG10 under both irrigation regimes, with  
values of 38.68 under 100% irrigation and 37.04 under 40% irrigation. Agronomic  
zinc use efficiency (AZUE) also reached its highest values in SRFHG10, at 11.00 and  
10.82 under 100% and 40% irrigation, respectively. In contrast, the Zn and Fe  
treatments showed negative AZUE values under both irrigation regimes, indicating weak  
agronomic performance relative to the control.  
Internal zinc utilization efficiency (IZUE) ranged from 10.22 to 12.23 under 100%  
irrigation and from 11.42 to 12.87 under 40% irrigation. The zinc harvest index (ZHI)  
ranged from 40.7% to 50.8% under 100% irrigation and from 42.5% to 53.9% under  
40% irrigation. Based on the target seed Zn concentration of 40 mg/kg, only  
SRFHG10 achieved the desired biofortification threshold under 100% irrigation.  
Discussion  
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The present findings indicate that cellulose-based SRFHG improved Zn accumulation  
in both seed and stover relative to non-hydrogel or less effective treatments. The  
advantage of SRFHG10 under both irrigation regimes is in line with the general role of  
controlled-release and water-retentive hydrogel systems in improving nutrient availability  
in the rhizosphere (Jiang et al., 2020; Peng et al., 2016; Tang et al., 2020). The  
biological importance of enhanced seed Zn concentration is substantial because Zn is  
essential for crop metabolism and is also a key micronutrient in human diets (Alloway,  
2008; Cakmak, 2008).  
Although Zn concentration improved in both plant fractions, the stover consistently  
accumulated more Zn than the seed. This pattern supports previous explanations that  
physiological barriers limit complete Zn remobilization to seeds, even when the external  
Zn supply is increased (Waters & Sankaran, 2011; White & Broadley, 2011). The  
strong irrigation response, especially in the stover, further supports the view that drought  
restricts nutrient mobility, uptake, and transport in mung bean and related crops  
(Ranawake et al., 2011; Rashid et al., 2003; Islam et al., 2021).  
The superior ZUE and AZUE values of SRFHG10 and SRFHG5 suggest that  
hydrogel-mediated Zn delivery improved nutrient use efficiency under both normal and  
moisture-stressed conditions. The negative AZUE values in the Zn and Fe treatments  
may indicate nutrient antagonism or ineffective Zn availability, which agrees with the  
broader concept of nutrient interaction effects in crop plants (Fageria, 2001; White &  
Broadley, 2009). Overall, these findings support the use of hydrogel-based  
micronutrient delivery as a practical strategy for improving Zn biofortification in mung  
beans, although further validation across seasons, soils, and genotypes is still needed.  
Conclusion  
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Cellulose-based slow-release hydrogel fertilizer substantially improved Zn  
biofortification of mung bean under both normal and drought-stress irrigation regimes.  
Among the evaluated treatments, SRFHG10 produced the highest Zn concentration in  
seeds and stover and showed the strongest Zn use efficiency. However, drought stress  
reduced seed Zn concentration and prevented most treatments from reaching the  
biofortification target of 40 mg/kg. These results demonstrate that hydrogel-mediated  
Zn delivery is a promising approach for improving micronutrient enrichment in mung  
bean; however, further optimization is required to achieve stable performance under  
water-limited conditions.  
Declarations  
Conflict of Interest: The authors declare no conflict of interest  
Funding: The authors gratefully acknowledge the Higher Education Commission  
(HEC) of Pakistan for financial support through the International Research Support  
Initiative Program (IRSIP), which facilitated the research work.  
Data availability: The data supporting the findings of this study are available from the  
corresponding author upon reasonable requests.  
Ethics approval: Not applicable for a plant-based pot experiment, unless required by the  
host institution.  
Tables  
Table 1. Nutrient sources and hydrogel loading were used for mung bean cultivation.  
Nutrient source  
Product (%)  
Field rate (lb/A)  
Amount added to  
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hydrogel (g/pot)  
ZnSO4  
5.6% Zn  
25  
0.08  
Table 2. Zinc concentration in mung bean seeds under normal and drought irrigation  
conditions.  
Treatment  
Control  
100% irrigation  
30.80  
40% irrigation  
29.40  
Mean  
30.10 g  
34.85 d  
35.77 c  
37.15 b  
42.67 a  
32.75 e  
31.00 f  
-
SRHG5  
SRHG10  
SRFHG5  
SRFHG10  
NPK-Fe,Zn  
Zn,Fe  
36.90  
32.80  
36.83  
34.70  
37.47  
36.83  
46.57  
38.77  
33.10  
32.40  
32.50  
29.50  
Mean  
36.31  
33.49  
Means followed by different letters differ significantly at p ≤ 0.05.  
Table 3. Zinc concentration in mung bean stover under normal and drought irrigation  
conditions.  
Treatment  
Control  
100% irrigation  
28.90  
40% irrigation  
25.80  
Mean  
27.35 g  
31.90 d  
SRHG5  
35.80  
28.00  
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SRHG10  
SRFHG5  
SRFHG10  
NPK-Fe,Zn  
Zn,Fe  
42.00  
54.60  
59.40  
34.20  
33.80  
41.24  
39.40  
49.80  
52.00  
29.10  
28.60  
36.10  
40.70 c  
52.20 b  
55.70 a  
31.65 e  
31.20 f  
-
Mean  
Means followed by different letters differ significantly at p ≤ 0.05.  
Table 4. Two-way ANOVA summary for zinc concentration in seeds and stover  
Variable  
Source of df  
variation  
SS  
MS  
F-value  
p-value  
Seed Zn  
Seed Zn  
Seed Zn  
Nutrient  
Irrigation  
N × I  
6
1
6
6
1
6
654.058  
83.726  
57.352  
109.010  
83.726  
9.559  
9.32  
0.00  
0.01  
0.57  
0.00  
0.00  
0.00  
7.16  
0.82  
Stover Zn Nutrient  
Stover Zn Irrigation  
Stover Zn N × I  
4537.556 756.259  
756.26  
277.71  
5.73  
277.714  
34.376  
277.714  
5.729  
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Note: The error mean square was 11.695 for seed Zn and 1.000 for stover Zn; the CV  
was 9.80% and 2.59%, respectively.  
Table 5. Zinc efficiency indices under normal and drought irrigation conditions.  
Treatment ZUE  
100%  
ZUE  
40%  
AZUE AZUE IZUE  
IZUE  
40%  
ZHI  
ZHI  
40%  
100%  
40%  
1.18  
7.86  
9.64  
10.82  
0.39  
100%  
11.07  
12.23  
11.66  
10.22  
11.87  
100%  
SRHG5  
28.75  
34.43  
38.36  
27.39  
34.07  
35.86  
37.04  
26.61  
1.07  
12.61  
12.87  
11.59  
11.42  
12.11  
50.8  
46.7  
40.7  
43.9  
49.2  
53.9  
46.8  
42.5  
42.7  
52.7  
SRHG10  
SRFHG5  
6.75  
10.68  
11.00  
0.86  
SRFHG10 38.68  
NPK-  
Fe,Zn  
28.54  
Zn,Fe  
26.55  
25.18  
-1.14  
-1.04  
11.21  
12.13  
49.0  
50.8  
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