Heliyon 9 (2023) e14964

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Heliyon
journal homepage: www.cell.com/heliyon

Research article

Investigating the effectiveness of LED lighting in the production of

rich sprouts for food purposes
Liang Wang a, Chen Sun a, Haiye Luan b, *, D.T. Semiroumi c
a
North Jiangsu Institute of Agricultural and Rural Modernization, Yancheng Teachers University, Jiangsu, 224007, China
b
School of Marine and Biological Engineering, Yancheng Teachers University, Jiangsu, 224007, China
c
School of Mechanic, Islamic Azad University, Khomeinishahr Branch, Isfahan, Iran

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

Keywords: In recent times, there has been a surge in interest in the impact of diet and physical activity on
Seed germination human health, with the goal of expanding lifespan and enhancing the quality of life. This has
Chlorophyll Light-emitting diode (LED) to interventions centered on incorporating healthy foods, including
Nutrition value
fresh sprouts, which are rich in antioxidant compounds and beneficial phytonutrients for human
Vitamin C
consumption. Various factors, such as temperature, nutritional solution, and types of light quality
Land resource utilization
and intensity, can influence the nutritional value of sprouts. This study evaluates the impact of
LED light with red-blue-ultraviolet (6:3:1; R:B:UV) and three levels of intensity (control, 120, and
150 μmol/m2s− 1) on five different sprout species, namely wheat, barley, mung bean, alfalfa, and
soybean, after seven days of germination. The research investigates the effects on various pa­
rameters, including photosynthetic pigments (chlorophylls a, b, total), carotenoid, activities of
antioxidant enzymes such as catalase, superoxide dismutase, and soluble proteins, soluble sugars,
starch, vitamin C, and element content such as potassium, iron, and phosphorus. The results
indicate that the LED treatments and increasing light intensity significantly improve the physi­
ological and antioxidant properties of edible sprouts, with the 150 μmol/m2s− 1 treatment pro­
ducing the most beneficial outcomes. Additionally, increasing light intensity reduces starch
content while enhancing the content of photosynthetic pigments, soluble carbohydrates, vitamin
C, element concentration, antioxidant enzymes, and soluble proteins. Among the five species of
edible sprouts, barley had the highest content of photosynthetic pigments, while soybean and
mung beans had the lowest content. Mung beans and alfalfa had the highest and lowest con­
centrations of potassium and iron, respectively. In terms of phosphorus concentration, soybean
and barley sprouts showed the highest and lowest concentrations, respectively.

1. Introduction

In every society, the national capital is regarded as a crucial component that contributes to the autonomy and durability of that
society’s existence. However, these invaluable resources, bestowed by the Almighty, are at risk due to the actions of human beings and
their utilization of natural resources. Hence, acquiring knowledge and comprehension of human necessities, predicaments, economic
and social quandaries can aid in comprehending the reasons for an unfavorable association with the environment and determining
effective solutions to mitigate its degradation [1–3]. In recent years, there has been an increasing interest in exploring the impact of

* Corresponding author.
E-mail addresses: Luanhy@yctu.edu.cn (H. Luan), davoodtoghraie@yahoo.com (D.T. Semiroumi).

https://doi.org/10.1016/j.heliyon.2023.e14964
Received 23 June 2022; Received in revised form 21 March 2023; Accepted 22 March 2023
Available online 31 March 2023
2405-8440/© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
L. Wang et al. Heliyon 9 (2023) e14964

physical activity on human health. Smart rooms constructed with innovative materials (fiber composite) can improve cultivation
conditions by controlling the mechanical behavior of light and environmental conditions [4–6]. This interest is driven by the desire to
enhance life expectancy and improve quality of life, leading to interventions that incorporate new healthy foods. These foods provide a
rich source of beneficial nutrients that can help delay the onset of chronic and debilitating diseases [7,8]. As consumers seek out foods
that promote health and well-being, a wide range of plants, products, and foods have been investigated to assess their nutritional and
chemical composition, with particular attention given to bioavailability and bioactivity [9–11]. One area of focus has been on diets
that are high in fruits and vegetables, including microgreens and small vegetables that fall under the category of edible sprouts.
Germination sprouts, which are fully grown seeds that have been sprouted in water over several days, are particularly rich in vitamins,
minerals, proteins, fats, and carbohydrates that are required for plant growth. The germination process leads to significant chemical
changes in these seeds, making them a valuable addition to a healthy diet [12–14].
Sprouts are considered biogenic food due to their high nutritional value, including vitamins, minerals, proteins, and other healthy
compounds. The growth period of sprouts varies from 3 to 14 days depending on the species. The use of seeds, grains, legumes, and
sprouted nuts dates back thousands of years. Bean sprouts are known for their health benefits in preventing and treating various
diseases [15–19]. Additionally, sprouts have been found to have anti-inflammatory properties, which can help with rheumatism and
have a laxative effect. Furthermore, sprouts are essential to the barley malting process, as the germination of seeds leads to significant
chemical changes [20–24]. Hence, sprouts are referred to as biogenic food, which is living food that contains numerous vitamins,
minerals, proteins, and other beneficial compounds [25–34].
In Eastern countries, edible sprouts have a long-standing culinary history, where seedlings are consumed as a vital ingredient. To
facilitate the germination process, Light-emitting diode (LED) lamps are utilized in growth chambers, providing light with varying
color characteristics, including white, red, blue, green, and their combinations [35–39]. The impact of LED on germination growth
varies based on the species. The use of LED technology is a promising and effective approach for producing sprouts, microgreens, and
hydroponic fodder with enhanced nutritional value. This research was conducted to investigate the impact of LED light intensities,
including red, blue, and ultraviolet, on edible sprouts. The study showed that LED treatments with increasing light intensity enhanced
the physiological and antioxidant properties of the edible sprouts, compared to the control treatment [35–38].

2. Materials and methods

2.1. Plant materials and growth conditions

The present study employed a factorial design, utilizing completely randomized blocks with two types of treatments and three
replications. The first treatment consisted of three levels of light intensity, including a control, and two intensities of 120 and 150
μmol/m2s− 1, achieved by utilizing red-blue-ultraviolet LEDs in a ratio of 6:3:1 R:B:UV. The second treatment consisted of five types of

Fig. 1. The amount of chlorophyll a content, (a) Chlorophyll b, (b) Total chlorophyll content, (c) Carotenoid content, (d) Under the treatment of
LED (control, 120 and 150 μmol/m2s− 1) and sprout (wheat, barley, mung bean, soybean, alfalfa).

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edible sprouts (wheat, barley, mung bean, soybean, and alfalfa) grown in a growth chamber with a sensor for temperature, humidity,
and pH, and LED light control. The growth chamber walls were made of 313 sheets of steel, and the cultivation shelves were con­
structed of coated aluminum with a distance of 30 cm between them. The LED lamps were positioned 30 cm away from the sprouting
medium. The study aimed to investigate the effect of different light intensities on plant seed yield and behavior.
The utilization of LED and SMD light may render the effect indistinguishable due to differences in consumption and time intervals.
The experiment lasted for 7 days, during which the sprouts were exposed to LED light for 16 h and kept in darkness for 8 h starting from
the second day. Additionally, the sprouts were uniformly irrigated four times daily using a mechanized irrigation system in the growth
chamber. On the third and sixth day, they were fertilized with complete fertilizer 20-20-20 (Bavaria company) with a concentration of
2 g per liter, twice a day. The average relative humidity was 65–70%, and the day and night temperature of the growth chamber was
maintained at 25 and 21 ◦ C, respectively. The seeds were obtained from national Agriculture company and planted after proper
germination. After 7 days, the sprouts were evaluated for various traits such as chlorophyll, carotenoids, vitamin C, antioxidant ca­
pacity, and nutrient content, and were measured using the prescribed methods.

2.2. Determination of the chlorophyll and carotenoid contents

In accordance with Lichtenthaler’s (1987) methodology [32], the quantification of chlorophyll and carotenoid content was carried
out as follows: 1 g of fresh sprouts was homogenized with 10 ml of absolute acetone, and the resulting mixture was centrifuged at 5000
rpm for a duration of 10 min. Using spectrophotometry, the absorbance of the photosynthetic pigments was measured at 645 and 662
nm, respectively, for chlorophyll a and b, and at 470 nm for carotenoids as shown in Fig. 1(a–d). The total content of chlorophyll and
carotenoids was determined using the Lichtenthaler method.

2.3. Soluble carbohydrates and starch measurements

To extract soluble carbohydrates from 0.2 g of leaf material, 2.5 ml of 80% ethanol was used at 90 ◦ C for a total of 60 min (in two
30-min steps). The resulting extracts were filtered and evaporated with alcohol. The resulting precipitate was dissolved in 2.5 ml of
distilled water, and 200 μl of each sample was mixed with 5 ml of Antron reagent in a test tube. After incubation at 90 ◦ C for 17 min and
cooling, absorbance was measured at 625 nm. The concentration of each sample was determined using a standard glucose curve in mg/
g fresh weight. To determine the content of insoluble sugars (starch), the Haissig method (1979) was used. Specifically, the dried
precipitate obtained from the alcohol extraction was mixed with approximately 40 ml of distilled water in a Falcon tube and boiled for
10 min. The contents were filtered, and the precipitates were returned to the Falcon and boiled again with 40 ml of distilled water for
10 min. In this experiment, a filtered solution comprising of 100 ml of distilled water was utilized to quantify insoluble sugars, using
the phenol sulfuric acid method. To do so, 0.5 ml of each extract was mixed with distilled water to increase the volume of the test tubes
to 2 ml. Then, 1 ml of 5% phenol was added and the mixture was agitated. This was followed by the addition of 5 ml of concentrated

Fig. 2. The amount of starch content (a) soluble carbohydrates, (b) vitamin c content, (c) under the treatment of LED (control, 120 and 150 μmol/
m2s− 1) and sprout (wheat, barley, mung bean, soybean, alfalfa).

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sulfuric acid to each tube, which was carried out under hydrated conditions due to the substantial heat generated during the reaction.
The absorbance of the samples was measured at 485 nm, and subsequently, the concentrations of insoluble and soluble sugars were
determined and expressed as milligrams per gram of dry weight as shown in Fig. 2(a–c).

2.4. Assessing the activity of antioxidant enzymes and soluble protein

In order to quantify protein levels, sprout samples were collected and subjected to extraction at a temperature of 4 ◦ C. Specifically,
1 g of fresh leaf tissue was homogenized in a porcelain mortar with 3 mL of 50 mM phosphate buffer (pH = 7.2) supplemented with 1
mM ethylene diamine tetra acetic acid (EDTA), 1 mM nylon methane sulfonyl fluoride (PMSF), and 1% polyvinyl pyrrolidone (PVP).
The resulting extract was then subjected to centrifugation in a refrigerated centrifuge at 14,000 rpm for 15 min at 4 ◦ C, with the
resulting transparent supernatant used for measuring both antioxidant enzymes and protein as shown in Fig. 3(a–c). The protein levels
were determined using Bradford’s (1976) [35] method, where 0.1 mL of protein extract were mixed with 5 mL of Biorea reagent in test
tubes and immediately analyzed. The specific activity of the catalase enzyme was measured by the modified Aebi (1974) method [36],
and by H2O2 decomposition spectrophotometry at 240 nm. Measurement of specific activity of superoxide dismutase enzyme was
measured using modified and modified Assady method (2011) [37].

2.5. Vitamin C measurement

The concentration of ascorbic acid, also known as vitamin C, was determined by employing the 2, 6-dichlorophenol method. In this
method, vitamin C reacts with the dye reagent dichlorophenol endophenol, which functions as an oxidation and reduction agent,
resulting in a colorless solution (Table 1). After completion of the reaction, the color detector in the acidic solution turned from non-
reduction to purple. Subsequently, 1 ml of the resulting solution was titrated with the 2, 6-dichlorophenol and endophenol reagent,
with the reagent being added dropwise until a pink color persisted for 15 s. The titration was then stopped, and the amount of ascorbic
acid was calculated in grams per liter and reported in milligrams, using a formula. The procedure was repeated three times, and the
concentration of vitamin C was determined using the following equation [38].
– NI × VI
NC × VC–

NC × VC=ND × VD

Mg C = 10-3 Li × (X mol/1Li) × (119.01/1 mol C) × (103 mg/1g) = 119.01 × N

The percentage of vitamin C in each sample was calculated using the following equation. The fresh weight of each sample was
considered 0.5 g.

Fig. 3. The activity of the antioxidant enzyme catalase, (a); Superoxide dismutase (SOD), (b); and protein content, (c); under the treatment of LED
(control, 120 and 150 μmol/m2s− 1) and sprouts (wheat, barley, mung bean, soybean, alfalfa).

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Table 1
Specifications of the sprouts used in the experiment.
Number sprouts name Scientific name

1 Wheat –
2 Barley Hordeum vulgare cv
3 Mung bean Vigna radiate cv. NM92
4 Soybean Glycine max cv. williams 82
5 Alfalfa Medicago sativa cv. ranger

C = (Mg C/sample weight) × 100

2.6. Content of an element

In terms of nutritional content and antioxidant activity, different types of sprouts were analyzed using various methods. Soybean
sprouts had the highest vitamin C content, while wheat sprouts had the lowest. Meanwhile, alfalfa and mung bean sprouts had the
highest and lowest levels of soluble carbohydrates, respectively. Wheat and soybean sprouts had the highest and lowest levels of starch,
respectively. The antioxidant enzyme catalase was found to be most active in alfalfa sprouts, while soybean sprouts had the least
activity. On the other hand, the highest and lowest levels of superoxide dismutase antioxidant activity were found in soybean and
barley sprouts, respectively. The levels of different elements were determined using Rana Munns’s method (2010) and measured via
the dry ash method. This involved burning organic matter in a porcelain plant at 550 ◦ C for 2 h and dissolving the ashes in hydrochloric
acid. The resulting solution was filtered and diluted before measuring the levels of soluble potassium, iron, and phosphate using
various instruments such as a flame photometer and atomic absorption spectrometers. The phosphate level was determined using the
Motsara and Roy Method (2008) and measured using a spectrophotometer at 880 nm.

2.7. Statistical analysis

In this study, the data was analyzed using a factorial experiment design with a completely randomized approach that involved three
replications. The statistical analysis was conducted using SAS software version 9.2, and the mean differences were assessed through the
application of Duncan’s multiple range tests with a confidence level of 95%.

3. Results and discussion

3.1. Photosynthetic pigments and vitamin C content

In order to enhance the aggregation of specific compounds, optical spectra can be obtained by selecting particular wavelengths, as
well as in combination with each other. The use of LED lights is known to promote the production of beneficial chemical compounds,
including phenolic compounds, vitamins, glycosylates, chlorophyll, and carotenoids. LED lights are also known to provide the
necessary photons for plant growth across various plant parts. Red-blue lights have been evaluated extensively in the scientific
community to determine their impact on plant growth, and studies have shown that plants grown under these lights can produce
products as healthy as those grown under full-spectrum light. However, the high cost of these lights has limited their commercial use,
particularly for large-fruited plants. Nonetheless, they can be justified for some crops, such as sprouts, small leafy vegetables, and
hydroponic forage that can grow under high-density conditions. Additionally, research has indicated that blue-red lights can influence
the chemical composition and quantity of secondary metabolites. Various alternatives to chemical fertilizers such as mycorrhizal fungi
and biochar are commonly used to improve crop growth in soil [16].
Ultrasound treatment was found to be useful in improving the quality of steam buns by controlling the inherent starch structure in
the dough [17]. Germination was observed to increase the level of bioactive compounds [18]. The pathogenic fungus, Aspergillus
flavus, responsible for contaminating agricultural products with aflatoxin, was prevented by paraformaldehyde by inducing additional
reactive oxygen species (ROS) [19]. The study introduced the antimicrobial properties of SEO and its main active components, while
cinnamaldehyde (CA) was found to exhibit antifungal activity against Fusarium solani [20]. Henna is the largest international buyer of
soybeans, accounting for approximately 66% of the global trade [21]. During the growth season, the biomass and morphology of fine
roots were significantly influenced [22]. Soil microorganisms play a crucial role in the underground carbon (C) cycle of terrestrial
ecosystems, contributing to a significant part of the genetic diversity [23]. Soil respiration was higher when organic nitrogen was
added than when inorganic nitrogen was added [24,25]. The proposed framework in Ref. [26] emphasizes the importance of plant and
soil properties in guiding microbial life history traits during rangeland restoration. It has been reported by certain researchers that the
incorporation of green light in the light composition has positive impacts, as evident from the experiments conducted with
red-blue-green lights [39]. Additionally, the use of other light spectra, such as far red, has been found to play a crucial role in plant
biological processes and photosynthesis. Moreover, ultraviolet light has been shown to enhance the expression of specific molecules in
plants [39–42]. The statistical analysis of variance revealed that the influence of LED lamp treatment and sprouts type on photo­
synthetic pigments, including chlorophyll-a, b, and total, was statistically significant at a significance level of 1% (Table 2). The results
illustrated that LED treatment with 150 μmol/m2s− 1 shows the greatest effect on increasing photosynthetic pigments and control

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treatment (dark condition) shows the lowest content of photosynthetic pigments (Tables 3 & 4). The results of the effect of edible
sprouts on photosynthetic pigments were significant at the level of 1% and barley sprouts demonstrated the highest number of
photosynthetic pigments and alfalfa sprouts had the lowest amount of chlorophyll a, soybean sprouts had the lowest amount of
chlorophyll b, and carotenoids and mung bean sprouts had the lowest concentration of total chlorophyll (Tables 5 and 6). According to
the analysis of the variance table, the interaction of LED * sprouts on chlorophyll a, b and total was significant at the level of 1%
(Table 2).
The maximum content of chlorophyll a, b, and total were measured from barley sprouts under light with an intensity of 150 μmol/
m2s− 1, and the minimum amount of chlorophyll a and total was obtained in control soybean sprouts and chlorophyll b in control mung
bean sprouts (Tables 7 & 8). Also, according to the analysis of variance table, the effect of light on carotenoid pigment was significant at
5% level and it was the most content in light intensity of 150 μmol/m2s− 1and the lowest was in the control treatment (Table 2). The
effect of sprout type on carotenoids was also significant at the 5% level so that the highest number of carotenoids was obtained in
barley, and the lowest amount was obtained in soybeans (Tables 6 and 7). Vitamin C: The results of the analysis of variance
demonstrated that the effect of LED lamp treatment and sprouts type on vitamin C content was significant at the level of 1% (Table 2).
Moreover, the interaction of LED light × sprout was significant on the content of vitamin C at the level of 1% and results show that
soybean sprouts under the light of 150 μmol/m2s− 1 had the most content of vitamin C and the lowest amount in control wheat sprouts
was obtained (Table 8).

3.2. Carbohydrates and starches

The results of the analysis of variance table indicated that the effect of LED lamp treatment and sprouts type on carbohydrate and
starch content was significant at the level of 1% (Tables 2 and 3). Also, the interaction of LED light × sprouts on the amount of
carbohydrate content at the level of 1% showed that the highest amount was measured in barley and wheat sprouts under 150 μmol/
m2s− 1 and the minimum amount was measured in mung bean and soybean sprouts (Table 8). The analysis of variance of starch
indicated that the interaction of LED light × sprout on the concentration of starch at the level of 1% was significant and its highest
value was in the wheat sprouts control treatment and its minimum value was in alfalfa sprout under LED light treatment with an
intensity of 180 μmol/m2s− 1 was observed (Table 9).

3.3. Antioxidant enzymes and proteins

The results of the analysis of variance showed that the effect of LED lamp treatment and sprout types on antioxidant enzymes
including catalase and superoxide dismutase and total protein was significant at a 1% level (Table 3). Additionally, the interaction of
LED light × the sprout was significant on the mentioned traits at the level of 1%, so that the most activity of catalase was measured in
barley sprouts treated with 150 μmol/m2s− 1 LED treatment, and the minimum activity was found in barley sprouts under control
treatment. The most and lowest levels of superoxide dismutase antioxidant enzyme were measured in soybean sprouts with 150 μmol/
m2s− 1 LED treatment and in wheat, barley, and mung bean sprouts, respectively as can be seen in Table 9. The comparison table of
mean protein showed that its maximum and minimum values were obtained in mung bean sprouts treated with 150 μmol/m2s− 1 LED
treatment and barley sprouts with control treatment, respectively (Table 9).

3.4. Content of mineral elements

According to the results of the analysis of variance table, LED lamp treatment and sprouts type on the content of potassium, iron,
and phosphorus minerals were significant at the level of 1% (Table 3). Also, the interaction of LED light in the sprouts was significant
on mineral elements at the level of 1% and the highest and the amount of potassium, iron, and phosphorus content in mung bean
sprouts under LED light treatment with the intensity of 150 μmol/m2s− 1 and the lowest content were measured in mung bean, barley
and mung bean under control treatment, respectively as shown in Table 9 and Fig. 4(a–c). Almost all plants need light throughout their
life cycle, from germination to flowering and eventually seed production [41]. Plants can respond to their non-living environment and
adapt their morphology and physiology to promote optimal growth and development [42]. Light is one of the most important envi­
ronmental factors and plays an important role in plant growth and physiology [43]. Because plants are unable to move, they depend on
environmental adaptation mechanisms and diverse responses to changes in impact, spectral quality, direction, and duration [42–44].

Table 2
Analysis of variance for photosynthetic pigments, vitamin c, and soluble carbohydrates subjected to three LED treatment (control, 120 and 150 μmol/
m2s− 1) and five sprout type treatment (wheat, barley, mung bean, soybean, alfalfa).
Source of df Chlorophyll a Chlorophyll b Chlorophyll Total Carotenoid Vitamin C Soluble Carbohydrates

Rep 2 0.018 ns 0.039 ns 0.030 ns 0.012 ns 0.041 ns 25.48 ns

LED 2 1.452** 0.025** 1.81** 0.042* 28.19** 21408**
sprout 4 0.067** 0.064** 0.077** 0.013 * 32.82** 442**
LED × sprout 8 0.003** 0.023** 0.040** 0.011ns 1.07** 72.87**
Error 28 0.002 0.013 0.037 0.01 0.025 17.53
CV 2.25 5.1 2.71 15.71 3.26 4.07

* ** andns significant at the level of probability of 5%, 1% and non-significant, respectively.

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Table 3
Analysis of variance for starch, antioxidant enzymes, protein and mineral content (potassium, iron, and phosphorus) subjected to three LED treatment
(control, 120 and 150 μmol/m2s− 1) and five sprout type treatment (wheat, barley, mung bean, soybean, alfalfa).
Source of df Starch Catalase Superoxide dismutase Protein potassium Iron phosphorus
ns ns ns ns ns ns
Rep 2 0.69 0.039 1.4 0.580 0.50 0.13 0.099 ns
LED 2 438.6** 196889** 34142** 356.7** 7649.3** 41.60** 310.67**
sprout 4 414.51** 580.7** 423** 20.03** 187.08** 28.5** 7.35**
LED × sprout 8 14.5** 458.9 ** 42.20** 0.667** 149.19** 5.13** 5.09**
Error 28 0.579 3.146 1.36 0.036 1.71 0.843 0.056
CV 4.24 0.94 2.71 1.52 1.64 11.07 1.91

*, ** and ns significant at the level of probability of 5%, 1% and non-significant, respectively.

Table 4
Mean comparison for photosynthetic pigments, vitamin C and soluble Carbohydrates under three LED treatment (control, 120 and 150 μmol/m2s− 1).
Chlorophyll a Chlorophyll b Chlorophyll Total Carotenoid Vitamin C Soluble Carbohydrates
1 1 1 1 1 1
Treatment mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grfw−
LED control 0.28c 0.032c 0.31c 0.026c 3.36c 61.13c
120 0.75b 0.067b 0.83b 0.046b 5.10b 112.07b
μmol/m2s1
150 0.86a 0.11a 0.97a 0.128a 6.06a 134.93a
μmol/m2s1

Table 5
Mean comparison for starch, antioxidant enzymes, protein and mineral content (potassium, iron, phosphorus) under three LED treatment (control,
120 and 150 μmol/m2s− 1).
Starch Catalase Superoxide dismutase Protein potassium Iron phosphorus
− 1 − 1 − 1 − 1 − 1 − 1 1
Treatment mg/grfw mg/grfw mg/grfw mg/grfw mg/grdw mg/kgdw mg/kgdw−
LED control 23.82a 56.86c 116.17c 6.95c 55.06c 6.54c 7.32c
120 16.80b 236.07b 186.07b 14.43b 91.4b 8.49b 14.04b
μmol/m2s1
150 13.183c 270.13a 207.37a 16.12a 96.46a 9.85a 16.00a
μmol/m2s1

Table 6
Mean comparison treatment for photosynthetic pigments, vitamin c, and soluble carbohydrates under five sprout treatments (wheat, barley, mung
bean, soybean, alfalfa).
Chlorophyll a Chlorophyll b Chlorophyll Total Carotenoid Vitamin C Soluble Carbohydrates
1 1 1 1 1 1
treatment mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grfw−
wheat 0.70b 0.065c 0.767b 0.048c 2.64d 109.22a
barley 0.75a 0.085a 0.835a 0.66a 2.95c 105.44a
sprout mung bean 0.53d 0.064c 0.595d 0.054b 6.57a 92.88c
soybean 0.60c 0.073b 0.676c 0.044c 6.47a 98.00b
alfalfa 0.62c 0.071b 0.673c 0.0484c 5.56b 108.00a

Table 7
Mean comparison for starch, antioxidant enzymes, protein and mineral content (potassium, iron, phosphorus) under five sprout treatments (wheat,
barley, mung bean, soybean, alfalfa).
Starch Catalase Superoxide dismutase Protein potassium Iron phosphorus
1 1 1 1 1 1 1
sprout treatment mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grdw− mg/kgdw− mg/kgdw−
wheat 25.75a 185.33d 165.38d 12.15c 84.27b 9.68b 11.96c
barley 20.42c 185.22c 163.33e 11.29d 79.88c 7.85c 11.28d
mung bean 21.77b 190.00b 167.39c 14.64a 86.44a 10.57a 12.75b
soybean 9.83e 176.33e 180.00a 13.34b 79.55c 6.88d 13.70a
alfalfa 11.89d 198.56a 173.58e 11.07b 74.72d 6.46d 12.57b

Light is one of the main factors affecting the growth and development of the plant. Light-receiving systems respond to the intensity
and quality of light, as well as to the duration and interruption of the process of metabolic pathways. In addition, light conditions may
cause changes in plants due to light, which leads to the function of the antioxidant defense system. As far as seed production is

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Table 8
Interaction effect of LED (control, 120 and 150 μmol/m2s− 1) and sprout (wheat, barley, mung bean, soybean, alfalfa) treatment for photosynthetic
pigments, vitamin C and soluble Carbohydrates.
treatment Chlorophyll a Chlorophyll b Chlorophyll Total Carotenoid Vitamin C Soluble Carbohydrates
− 1 − 1 − 1 − 1 − 1 1
LED Sprouts mg/grfw mg/grfw mg/grfw mg/grfw mg/grfw mg/grfw−
Control wheat 0.336g 0.038g 0.375j 0.051b 1.666j 68.00h
barley 0.350g 0.041g 0.391j 0.065b 2.033i 65.00h
mung bean 0.256h 0.027hi 0.284k 0.055b 4.866e 52.66i
soybean 0.207i 0.032h 0.239l 0.052b 4.3f 50.61i
alfalfa 0.270h 0.024i 0.294k 0.049b 3.933g 69.33h
120 μmol/m2s1 wheat 0.843c 0.058f 0.902e 0.061b 2. 3h 119.33de
barley 0.925b 0.085d 1.010b 0.055b 2.533h 113.67ef
mung bean 0.740d 0.066e 0.806g 0.053b 7.2c 104.67g
soybean 0.643f 0.057f 0.7i 0.050b 7.1c 113.33ef
alfalfa 0.683e 0.069e 0.753h 0.053b 6.4d 109.33fg
150 μmol/m2s1 wheat 0.926b 0.098c 1.025b 0.088b 3.966g 140.33ab
barley 0.975a 0.128a 1.104a 0.157a 4.3f 137.67b
mung bean 0.830c 0.126a 0.941d 0.056b 7.666b 121.33d
soybean 0.743d 0.103c 0.846f 0.074b 8.020a 130.00c
alfalfa 0.853c 0.119b 0.973c 0.077b 6.366d 145.33a

Table 9
Interaction effect of LED (control, 120 and 150 μmol/m2s− 1) and sprout (wheat, barley, mung bean, soybean, alfalfa) treatment for starch, antioxidant
enzymes, protein and mineral content (potassium, iron, phosphorus).
Treatment Starch Catalase Superoxide dismutase Protein potassium Iron phosphorus
1 1 1 1 1 1 1
LED Sprouts mg/grfw− mg/grfw− mg/grfw− mg/grfw− mg/grdw− mg/kgdw− mg/kgdw−
control wheat 34.366a 59.651k 111.672l 6.30h 55.00h 8.138def 7.33ij
barley 25.800c 45.250l 113.72l 5.98h 51.66i 5.238h 7.13j
mung bean 29.666b 245.33f 110.87l 9.23f 50.00i 7.4efg 6.36k
soybean 13.039g 66.33j 127.00j 7.96g 62.5g 6.566h 7.53i
alfalfa 16.237f 59.651k 119.00k 5.34i 56.16h 5.378h 8.26h
120 μmol/m2s1 wheat 23.462d 231.00h 179.00h 14.36e 95.83c 10.42b 13.2f
barley 18.530e 235.39g 175.00i 13.40e 91.69d 8.883bce 11.26g
mung bean 19.671e 245.35f 186.008 16.3c 101.33b 10.00bc 14.4e
soybean 9.433h 215.67i 200.00e 14.56e 87.16e 6.6fgh 16.23b
alfalfa 12.937g 253.00e 190.46f 13.53e 81.00f 6.6fgh 15.03d
150 μmol/m2s1 wheat 19.430e 275.00c 204.50d 15.8d 102.00b 10.50b 15.26cd
barley 16.930f 284.00a 201.31e 14.50e 96.36c 9.5bcd 15.46c
mung bean 16.00f 265.00d 207.00c 18.4a 108.00a 14.34a 17.5a
soybean 7.035h 247.00f 213.00a 17.5b 89.00e 7.5efg 17.35a
alfalfa 6.516h 279.67b 211.00b 14.40e 87.00e 7.434efg 14.43e

concerned, the use of LEDs during seed germination seems to be the most important strategy to improve the nutritional quality of
seedlings. The light intensity of LEDs has a significant effect on the content of chlorophyll a, b, total, and carotenoids, and with
increasing light intensity, their content also increases. In previous research, it has been demonstrated that exposure to blue light leads
to the accumulation of chlorophyll and other photosynthetic pigments. This is due to the fact that photoreceptors, including cryp­
tochrome and phototropin, receive blue light and closely related wavelengths, which then influence chlorophyll production in plants,
ultimately increasing its content. In a study involving 14-day-old sprouts of two rice cultivars with either purple or green leaves,
exposure to blue light resulted in increased levels of chlorophyll a and b, carotenoids, and total proteins within the leaves. Conversely,
the highest levels of anthocyanin in seedling leaves occurred when exposed to LED irradiation consisting of red and blue wavelengths
[45].
The highest accumulation of total carotenoids has been reported in dark buckwheat sprouts under the white light ratio [40–44].
The effect of natural light and LED on hydroponic forage of wheat showed that LED light with the intensity of 185 μmol/m2s− 1 with the
combination (60% red and 40% blue light) compared to natural light increased photosynthetic pigments that play an effective role in
the photosynthesis of plants [41–44]. Some studies have evaluated the different effects of LED treatment on carbon and nitrogen
metabolism and pigment concentrations. Wheat and barley grass (height 7–9 cm) were compared under control (high-pressure sodium
lamps and LED, 595 nm), supplemental LED light increased 1.6 and 1.3 times the amount of fructose and glucose in wheat seedlings
and also Increased glucose content and pigment cycle concentrations of xanthophylls (e.g, neoxanthin, violaxanthin, and zeaxanthin)
in the barley [42–45]. When the seed matures, the plant provides the nutrients the fetus needs in the form of starches, carbohydrates,
and complex proteins for the fetus to grow further. With the onset of the germination process and the onset of activity of the
alpha-amylase, beta-amylase germination enzymes, these substances are converted into simpler molecules and delivered to the fetus
for nutrition. Therefore, during germination, the amount of starch decreases, and the amount of carbohydrates increases. As the in­
tensity of light increases, the activity of enzymes involved in germination increases. Therefore, in sprouts located in the dark, the

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L. Wang et al. Heliyon 9 (2023) e14964

Fig. 4. The amount of Potassium content, (a); Iron (Fe), (b); Phosphorus content, (c); under the treatment of LED (control, 120 and 150 μmol/
m2s− 1) and sprout (wheat, barley, mung bean, soybean, alfalfa).

activity of these enzymes is less, and starch is decomposed to a lesser extent for cellular metabolism required for germination. On the
other hand, because the sprouts need to synthesize new proteins for growth, the complex proteins stored in the seed are broken down
and used to synthesize new proteins. Hence, the amount of soluble protein increases.
The obtained results of the study showed that light treatment and its intensity are effective on secondary compounds and in creating
the antioxidant capacity of different sprouts. During germination, increased activity of antioxidant enzymes such as catalase, super­
oxide dismutase, and other enzymes eliminates and inactivates a variety of reactive oxygen species. These enzymes reduce the per­
oxidation of lipids in the germination process, thereby increasing the germination rate and better germination growth. Plants are
equipped with antioxidant defenses such as superoxide dismutase and catalase enzymes to protect against ROS. In general, the activity
of antioxidant enzymes increasing during germination conditions. The effect of single wavelength under different light conditions on
radical scavenging activity, total phenol content, and interactions of other antioxidants during seed germination is still unclear. This is
probably due to seed sensitivity, and the severity of the antioxidant compounds in the tissues is related to seed genetics. On the other
hand, supplemental LED light (595 nm) with green light significantly affected the phenolic and vitamin C content in wheat and barley
sprouts, and red light significantly increased alpha-tocopherol in 3-day wheat sprouts. Also, the effect of radiation (UV–B more than
300 nm) on buckwheat sprouts was associated with increased anthocyanin, rutin, DPPH (2, 2-diphenyl-1-picryl-hydroxyl-hydrate),
and decreased radical oxygen species. Exposure of wheat sprouts to periods of natural and light compared to growing in complete
darkness has been studied and the highest amount of vitamin C, routine and free amino acids have been obtained. In a study on
hydroponic barley, the growth and dry matter were found to increase significantly when exposed to LED light with intensities of 160
and 220 μmol/m2s− 1 for 20 h with a relative humidity of 70%, compared to other light treatments.
This increase in growth characteristics was attributed to the role of LED light in completing the cycle of secondary metabolites, such
as phenolic compounds and vitamin C, resulting in an increase in the amount of vitamin C as the intensity of LED light was enhanced.
Additionally, growth under LED light was found to increase the antioxidant capacity, as well as the accumulation of mineral elements
such as calcium, potassium, iron, and zinc as shown in Fig. 4(a–c). Compared to conventional light sources, LED light is an economical,
cool, and controllable light source that can selectively and quantitatively provide different light spectra. In the study, the impact of
different light regimes on the nutritional properties of various sprouts was investigated. For instance, red + blue LEDs were found to
have no significant effect on the amino acid content of barley seedlings when compared to control plants grown under natural sunlight.
In addition, exposure of mung bean sprouts to low relative humidity and LED light with a wavelength of 600 nm led to improved
antioxidant properties and nutritional value.
LED light with blue and red wavelengths resulted in higher protein and chlorophyll content in rice sprouts, while a combination of
red, blue, and green light led to higher levels of antioxidants and vitamin C in wheat germ, radish, and lentil sprouts. Furthermore, LED
treatment was found to enhance nutrient uptake in the germination tissues of broccoli sprouts. Macronutrient concentration was
primarily affected by light treatment, with phosphorus concentration being higher in the 20 blue/80 red treatment, and magnesium
levels showing variance between lighting methods. These findings suggest that LED light can be used as an effective alternative to
natural sunlight for enhancing the nutritional properties of sprouts.

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L. Wang et al. Heliyon 9 (2023) e14964

4. Conclusion

In the course of their growth cycle, from germination to seed production, almost all plants require light and can respond to non-
living environmental factors to promote optimal development. Light is a key environmental factor and plays an important role in plant
growth, as plants cannot move and must rely on environmental adaptation mechanisms and diverse responses to changes in light
impact, quality, intensity, and duration. This study investigates the effects of LED treatments and increasing light intensity on the
physiological and antioxidant properties of edible sprouts, with treatment at 150 μmol/m2s− 1 significantly improving these traits
compared to treatment at 120 μmol/m2s− 1. Increasing light intensity of the LED treatments enhances the content of photosynthetic
pigments, soluble carbohydrates, vitamin C, element content, antioxidant enzymes, and soluble protein, while reducing the amount of
starch. Different types of sprouts exhibit unique physiological responses to light intensity, and increasing light intensity improves the
nutritional properties and element content of all sprouts. Plant tissues contain various pigments and photoactive compounds that
require energy in specific wavelengths to operate, and a comprehensive understanding of the mechanisms of these optical receptors,
the wavelengths received, and their effects on plant development and physiology is crucial for developing complementary optimal
methods for LED lighting, with emphasis on spectral quality and enhancing secondary metabolite production.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to
influence the work reported in this paper.

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