1

1 Article
2 Enhancing Growth and Glucosinolate Accumulation
3 in Watercress (Nasturtium officinale L.) by
4 Regulating Light Intensity and Photoperiod in a
5 Plant Factories
6 Vu Phong Lam1,2, Jaeyun Choi1 and Jongseok Park1,3*

7 1
Department of Horticultural Science, Chungnam National University, Daejeon 34134, Korea;
8 2
Department of Agronomy, Tay Bac University, Son La 360000, Vietnam
9 3
Graduate school of Bio-AI Convergence, Chungnam National University, Daejeon 34134,
10 Korea
11 * Correspondence: author’s: Jongseok Park, +82-42-821-5737, [email protected]

12 Abstract: Recent advancements in light-emitting diode technology provide a perfect

13 opportunity to evaluate the correlation between different light sources and plant
14 growth as well as their secondary metabolites. The aim of this study was to
15 determine the optimal light intensity and photoperiod for increasing plant growth
16 and glucosinolate concentration and content in watercress. Two-week-old seedlings
17 were transplanted in a semi-deep flow technique system of a plant factory for three
18 Citation: Lastname, F.; Last- weeks under four photoperiod–light intensity treatments (12 h-266 µmol .m−2.s−1, 16 h-
19 name, F.; Lastname, F. Title. 200 µmol.m−2.s−1, 20 h-160 µmol.m−2.s−1, and 24 h-133 µmol.m−2.s−1) with same daily
20 Agriculture 2021, 11, x. light integral. Shoot fresh and dry weights were the greatest under the 20 h-160
21 https://doi.org/10.3390/xxxxx µmol.m−2.s−1 treatment. Net photosynthesis and stomatal conductance gradually
22 decreased with decreasing light intensity and increasing photoperiod. However, total
23 Academic Editor: Firstname glucosinolate concentration increased with decreasing light intensity and increasing
24 Lastname photoperiod. The total glucosinolate content was the greatest under 20 h-160
25 µmol.m−2.s−1 treatment, because it depends on the shoot dry weight. These data
Received: date
26 suggest that the 20 h-160 µmol.m−2.s−1 treatment promoted the maximum shoot
Accepted: date
27 biomass and glucosinolate content in watercress. This study supplies the optimal
Published: date
28 light strategies for future industrial large-watercress cultivation.
Publisher’s Note: MDPI stays
29 neutral with regard to jurisdic-
Keywords: deep flow technique; glucosinolate, light-emitting diode; net
30 tional claims in published maps photosynthesis; shoot biomass, watercress
31 and institutional affiliations.

Copyright: © 2021 by the au-

thors. Submitted for possible
open access publication under
the terms and conditions of the
Creative Commons Attribution
(CC BY) license
(https://creativecommons.org/li
censes/by/4.0/).

2
3 Agriculture 2021, 11, x. https://doi.org/10.3390/xxxxx www.mdpi.com/journal/agriculture
 4 Agriculture 2021, 11, x FOR PEER REVIEW 2 of 14
5

32 1. Introduction
33 Watercress (Nasturtium officinale L.; Brassicaceae) is a semi-aquatic or
34 aquatic perennial herb mainly cultivated in Asia, North and South America,
35 and Europe [1]. Watercress is evaluated as an aquatic weed in some regions.
36 It is used in soups (as garnish), fresh salads, and in other dishes [2]. The
37 European Food Safety Authority has indicated that watercress belongs to
38 safe vegetable of the group “herbs, edible flowers, and leaf vegetables” [3].
39 The US Centers for Disease Control and Prevention selected the watercress
40 as one of the crops containing the highest nutrient content per calorie [4]. It
41 contains compounds such as vitamins, polyphenols, carotenoids, and
42 isothiocyanates; glucosinolates are the most crucial components present in
43 watercress [2]. Watercress is a known medicine for treating cough, ronchial
44 problems, and asthma [5]. Watercress has pharmacological actions such as
45 antioxidant, anti-inflammatory, cardioprotective actions, antipsoriatic,
46 antibacterial, and anticancer [1, 2, 6, 7]. Because of the abundance of
47 chemical components, watercress can use for food, medicine, and cosmetics
48 industries.
49 Growing plants in a plant factory using artificial light is an efficient
50 method of agricultural cultivation for combating climate change and
51 worldwide food shortage [8]. Water shortages, unusual weather, and
52 depletion of the agricultural land area result in a decrease in field crop
53 production globally [9, 10]. Nevertheless, these environmental problems do
54 not affect crop production in a closed plant factory as the conditions for
55 growth, are controlled using temperature regulation, air-condition, air
56 circulation fans, artificial light, nutrient solutions, and CO 2 enrichment [11,
57 12]. Crops in a plant factory always depend on artificial light (light intensity,
58 photoperiod, and light quality) which controls the photosynthetic process,
59 plant physiology, biochemistry, and morphology [13, 14]. Thus, upgrading
60 the light source efficiency would significantly decrease the expense of the
61 plant factory system, which in turn would promote their sustainable
62 cultivation, because the impacts of ecological and costs could be decreased.
63 The lighting can be supplied for plants at any time in a plant factory
64 because lighting schedules can be controlled, and particular photoperiods
65 can be selected to promote plant growth and quality. For example, longer
66 photoperiods increased the fresh weight of lettuce [15]. The growth of
67 lettuce was increased with longer photoperiods and lower photosynthetic
68 photon flux density (PPFD) at the same daily light integral (DLI), because the
69 longer photoperiods compensate for a lower PPFD [16]. The growth of
70 Achimenes cultivars grown under a low light intensity and longer
71 photoperiods was higher as compared that of those grown under a high light
72 intensity and shorter photoperiods at the same DLI [17]. In addition, plant
73 growth and morphology changes were reported due to changes in light
74 intensity and photoperiod [18]. In general, these reports indicated that plant
75 growth can be promoted under longer photoperiods with the same DLI.
76 There have been several studies on the influences of different light
77 intensities [19-21] and a combination of photoperiods and light intensities on
 6 Agriculture 2021, 11, x FOR PEER REVIEW 3 of 14
7

78 plant biomass and secondary metabolites [15, 22, 23]. The optimum plant
79 growth and quality can be obtained by controlling the light-emitting diode
80 photoperiod and light quality [24, 25]. Therefore, it is very important to
81 investigate the optimal light intensity and light photoperiod for plants before
82 cultivating them in a plant factory.
83 To date, several studies have been conducted on the influence of
84 different light qualities, photoperiods, and light intensities on the growth
85 and quality of watercress [26, 27]. However, the effects of different light
86 intensities in combination with different photoperiods on the growth and
87 glucosinolate concentration and content of watercress grown in a plant
88 factory have not yet been reported. Thus, the aim of this study was to find
89 the optimal light intensity and photoperiod treatment to increase plant
90 growth and glucosinolate content in watercress.

91 2. Materials and Methods

92 2.1. Seedling conditions
93 Watercress seeds were sown in rockwool cubes (240 holes; UR
94 Rockwool, Suwon, Korea) and grown in a plant factory for 2 weeks. The air
95 temperature and relative humidity in the plant factory were controlled at 20
96 ± 2 °C and 60 ± 10%, respectively. White fluorescent lamps (TL5 14W/865
97 Philips, Amsterdam, Netherlands) were used for illumination. The
98 photoperiod and PPFD were adjusted at 16 h per day and 150 µmol .m−2.s−1,
99 respectively. The Hoagland solution for watercress seedlings (electrical
100 conductivity 0.8 dS.m−1; pH 6.0) was supplied from 1 week after sowing.

101 2.2. Treatments

102 Two weeks after sowing, the seedlings were transplanted into four
103 lighting treatments in a plant factory with the same daily light integral
104 (11.52 mol‧m−2‧d−1). Each treatment had 10 plants. Treatment 12 h-266
105 µmol: Photoperiod was set to 12 h per day with the PPFD of 266
106 µmol.m−2.s−1. Treatment 16 h-200 µmol: Photoperiod was set to 16 h per day
107 with the PPFD of 200 µmol.m−2.s−1. Treatment 20 h-160 µmol: Photoperiod
108 was set to 20 h per day with the PPFD of 160 µmol .m−2.s−1. Treatment 24 h-
109 133 µmol: Photoperiod was set to 24 h per day with the PPFD of 133
110 µmol.m−2.s−1. Plants were cultivated under a 7:3 ratio of red:blue light-
111 emitting diodes (LEDs) for 28 days under 400 µmol .mol−1 CO2 concentration
112 and 60% ± 10% relative humidity. The blue and red LEDs had a peak
113 wavelength of 450 nm and 660 nm, respectively. The experiment was
114 conducted with two replicates for each treatment. The Hoagland nutrient
115 solution for watercress plants (EC 2.0 dS.m−1; pH 6.0) was supplied for 28
116 days. The day and night air temperatures were controlled at 22 and 20 °C,
117 respectively.

118 2.3. Measurement of plant growth parameters and SPAD value

 8 Agriculture 2021, 11, x FOR PEER REVIEW 4 of 14
9

119 After 28 days of transplanting, the shoot fresh and dry weights and stem
120 length were measured. The stem length and shoot fresh weight were
121 determined using a ruler and an electronic scale (EW220-3NM, Kern & Sohn
122 GmbH., Balingen, Germany), respectively. For determination of the shoot
123 dry weight, samples were dried for one week in an oven (HB-502M; Hanback
124 Sci, Suwon, Korea) at 70 °C and then weighed. The SPAD values were
125 measured with a portable chlorophyll meter (502, Minolta Camera Co., Ltd.,
126 Tokyo, Japan). All parameters were recorded for 6 plants (n = 6) in each
127 replication.

128 2.4. Leaf gas exchange parameters

129 The net photosynthetic rate and stomatal conductance of a fully
130 expanded leaves were measured with a portable photosynthesis system (LI‐
131 6400; Li‐Cor, Lincoln, NE, USA) at 28 days after transplantation. The
132 measurement conditions in the leaf chamber, namely CO 2 concentration, leaf
133 temperature, airflow rate, and PPFD, were maintained at 400 µmol·mol −1, 25
134 °C, 500 cm3·s−1, and 500 μmol m−2.s−1, respectively.

135 2.5. Determination of the individual glucosinolate concentration in

136 watercress
137 The individual glucosinolate concentrations in the watercress were
138 analyzed according to previous literature [28], but with some modifications.
139 The shoots of watercress plants were collected at 28 days after
140 transplanting, kept in a deep freezer at −70 °C after soaking in liquid
141 nitrogen, then moved to a dry freezer (TFD5503, IL Shinbiobase Co., Ltd.,
142 Korea) at -50 °C for 3 days, and ground to powder. Glucosinolate was
143 extracted in 70% (v/v) methanol with watercress powder (0.1 g) in a water
144 bath for 5 min. Afterward, the samples were centrifuged at 12,000 × g for 10
145 min, the supernatant was analyzed as described by Lam et al. 2019 and
146 Cuong et al. 2019, to determine individual glucosinolate concentrations
147 (glucobrassicin, 4-methoxyglucobrassicin, glucohirsutin, glucosiberin, and
148 gluconasturtiin; Table 1) [28, 29]. The individual glucosinolates were
149 measured by response factors (ISO 9167-1, 1992) and the HPLC peak area
150 ratios and with reference to a desulfo-sinigrin external standard [30].
151 Glucosinolate contents (µmol/ plant DW) were presented as total
152 glucosinolate concentration in the shoot (µmol g . -1
DW) multiplied by shoot
153 dry weight (g).
 10 Agriculture 2021, 11, x FOR PEER REVIEW 5 of 14
11

154 Table 1. Relative response factor values of the desulfo-glucosinolates from

155 watercress shoot extracts and their retention times on the C18 column.

Response fac- Retention

Side chain structure Common name.
tors times (min)
0.29 17.16 indol-3-ylmethyl Glucobrassicin
4-methoxyindol-3-yl- 4- Methoxyglucobras-
0.25 16.05
methyl sicin
1.1 13.78 8-methylsulfinyloctyl Glucohirsutin
1 16.77 7-methylsulfinylheptyl Glucosiberin
0.95 15.68 2-phenylethyl Gluconasturtiin

156 2.6. Statistical analysis

157 The growth and photosynthetic parameters were measured for six and
158 four plants per replication, respectively. The individual glucosinolate
159 concentrations were measured for three plants per replication. For
160 statistical analysis, one-way ANOVA was performed using SPSS 20.0 (SPSS,
161 Inc., Chicago, IL, USA). Significant differences among treatments were
162 verified at p ≤ 0.05, using Tukey’s multiple range test.

163 3. Results and discussion

164 3.1. Plant growth parameters, chlorophyll content, and photosynthetic
165 parameters
166 The 12 h-266 µmol.m−2.s−1, 16 h-200 µmol.m−2.s−1, and 20 h-160
167 µmol.m−2.s−1 treatments resulted in a higher growth parameters relative to
168 the 24 h-133 µmol.m−2.s−1 treatment. The shoot fresh and dry weights were
169 significantly higher (1.15- and 1.42-time, respectively) under the 20 h-160
170 µmol treatment than under the 24 h-133 µmol treatment (Fig. 1C and 1D).
171 However, the stem length and SPAD value of the watercress were not
172 significantly affected by photoperiod and light intensity combinations (Fig.
173 1A and 1B). Previous reports have indicated that higher light intensities
174 enhanced growth and promoted crop production [31, 32]. This was possible
175 because of the wider expansion of leaf under the higher light intensity
176 treatment. The bigger leaf permits more light interception, which might
177 have resulted in a significant increment in the shoot fresh and dry weights
178 under higher light intensity [31]. The growth of ice plants under a
179 fluorescent lamp, red LEDs, and blue LEDs was significantly higher under a
180 higher light intensity treatment (150 µmol.m−2.s−1) than under a lower light
181 intensity treatment (120 µmol.m−2.s−1) [20]. The biomass, stem diameter, and
182 root: shoot ratio of soybean were higher under 400 and 500 µmol .m−2.s−1
183 than under 100 µmol.m−2.s−1 [33]. The leaf area was reduced by shade
184 conditions [34]. Similarly, plant dry matter production of soybean decreased
185 with decreasing light intensity [35]. Leaf fresh weight of Arabidopsis
186 thaliana was significantly higher under a higher light intensity as compared
187 to that of plants grown under a low light intensity at 6 weeks after
188 transplanting [36]. Moreover, the fresh weight of watercress under a long
 12 Agriculture 2021, 11, x FOR PEER REVIEW 6 of 14
13

189 day (16 h) was significantly higher than that under a short day (8 h) [26].
190 The fresh and dry weights of quinoa increased under a short photoperiod
191 and high light intensity treatment [37]. Therefore, the biomass of the
192 watercress increased under high light intensity and short photoperiod
193 treatments.

194 .
195 Figure 1. Stem length (A), SPAD value (B), shoot fresh weight (C), and shoot dry weight (D)
196 under different lighting treatments of 12 h-266 µmol .m−2.s−1, 16 h-200 µmol.m−2.s−1, 20 h-160
197 µmol.m−2.s−1, and 24 h-133 µmol.m−2.s−1. Different letters above bar show significant
198 differences at p ≤ 0.05, using Tukey’s multiple range test (n = 6).

199 The net photosynthesis and stomatal conductance were reduced with
200 increasing photoperiod and decreasing light intensity (Fig. 2). Reductions of
201 light intensity may influence the carbon balance in the plant. Rates of
202 physiological process increases, while the photosynthetic yield decreases
203 [33]. Normally, it is expected that the shading conditions or lower light
204 intensities restrict leaf growth, and result in, smaller leaf areas with thinner
 14 Agriculture 2021, 11, x FOR PEER REVIEW 7 of 14
15

205 leaves, reduced chlorophyll content, and thinner palisade tissues, leading to
206 lower light-harvesting [38, 39]. Furthermore, there is a reduction in stomatal
207 conductance and density, which leads to poor CO 2 transportation under low
208 light conditions. The electron transition from PSII to PSI is obstructed,
209 whereas the activity and number of enzymes that participate in the Calvin
210 cycle undergo a change. All of this results in a reduced carbon dioxide
211 assimilation rate and a reduced net photosynthetic rate under low light
212 conditions [33]. Previous reports have indicated that the main biochemical
213 restraint related to shadow-associated down-adjustment of net
214 photosynthetic rate is a decrease in the activity or amount of Rubisco [33,
215 40]. Photosynthetic capacity was reduced under low light conditions because
216 carbon would be restricted [41]. For example, low light intensity reduced the
217 photosynthesis rate in pakchoi [42] and soybean [33]. Likewise, the net
218 photosynthetic under 16 h-200 µmol, 20 h-160 µmol, and 24 h-133 µmol
219 treatments was 1.38-, 1.52-, and 3.32-time lower than that of 12-266 µmol
220 treatment in the study, respectively. the stomatal conductance under 16 h-
221 200 µmol, 20 h-160 µmol, and 24 h-133 µmol treatments was 1.41-, 1.92-,
222 and 2.08-time lower than that of 12-266 µmol treatment in this study,
223 respectively. (Fig. 2). Moreover, the shoot fresh and dry weights of the
224 watercress were significantly lowest under lower light intensity and longer
225 photoperiod treatments (24 h-133 µmol) compared with 12-266 µmol,16 h-
226 200 µmol, and 20 h-160 µmol treatments. There was no significant
227 difference in shoot fresh and dry weights among 12-266 µmol,16 h-200 µmol,
228 and 20 h-160 µmol treatments (Fig. 1).

229
230 Fig. 2. The net photosynthesis (A) and stomatal conductance (B) under different lighting
231 treatments of 12 h-266 µmol.m−2.s−1, 16 h-200 µmol.m−2.s−1, 20 h-160 µmol.m−2.s−1, and 24 h-
232 133 µmol.m−2.s−1. Different letters above bars show significant differences at p ≤ 0.05, using
233 Tukey’s multiple range test (n = 4).

234 3.2. Total glucosinolate concentration and content

 16 Agriculture 2021, 11, x FOR PEER REVIEW 8 of 14
17

235 This analyses indicate that the watercress contained five different
236 desulfoglucosinolates (glucohirsutin, 4-methoxyglucobrassicin, glucoiberin,
237 glucohirsutin, and gluconasturtiin). Among the five desulfoglucosinolates
238 identified, gluconasturtiin presented the highest concentration (Table 2).
239 Gluconasturtiin accumulation in the shoot increased under 24 h-133 µmol
240 treatment and had the highest concentration (82.51% of the total
241 glucosinolate concentration). However, there was no significant difference in
242 gluconasturtiin concentration among 24 h-133 µmol, 16 h-200 µmol, and 20
243 h-160 µmol treatments and also between 20 h-160 µmol and 12 h-266 µmol
244 treatments. The highest glucosiberin, 4-methoxyglucobrassicin, and
245 glucohirsutin concentrations (13.61%, 3.79%, and 2.39% of the total
246 glucosinolates, respectively) were observed under 20 h-160 µmol treatment.
247 There was no significant difference in glucosiberin concentration among 24
248 h-133 µmol, 16 h-200 µmol, and 12 h-266 µmol treatments. There was no
249 significant difference in glucohirsutin concentration among four treatments
250 (24 h-133 µmol, 16 h-200 µmol, 12 h-266 µmol, and 20 h-160 µmol). There
251 was no significant difference in 4-methoxyglucobrassicin concentration
252 among 24 h-133 µmol, 16 h-200 µmol, and 20 h-160 µmol treatments and
253 also among 24 h-133 µmol, 16 h-200 µmol, and 12 h-266 µmol treatments.
254 The highest glucobrassicin concentration (8.80% of the total glucosinolate)
255 was recorded under 24 h-133 µmol treatment. There was no significant
256 difference in glucobrassicin concentration among 24 h-133 µmol, 16 h-200
257 µmol, and 12 h-266 µmol treatments (Table 2). Overall, the total
258 glucosinolate concentration was the greatest at 24 h-133 µmol treatment
259 and was 1.28-fold higher than that of the 12 h-266 µmol treatment. There
260 were no significant differences in total glucosinolate concentration in shoot
261 between the 12 h and 16 h treatments and also 20 and 24 h treatments,
262 respectively (Fig 3A). However, the total glucosinolate content in the shoot
263 was the highest under 20 h-160 µmol treatment, because glucosinolate
264 contents (µmol/ plant DW) were presented as total glucosinolate concentra-
265 tion in the shoot (µmol.g-1 DW) multiplied by shoot dry weight (g). There
266 were no significant differences in total glucosinolate content in shoot among
267 24 h-133 µmol, 16 h-200 µmol, and 12 h-266 µmol treatments (Fig 3B).
268 Glucosinolates are bioactive compounds typically in cruciferous group
269 plants. It has been indicated that long days could enhance glucosinolate
270 accumulation in Arabidopsis ([43] and watercress [26]. Low light intensity
271 increased the concentrations of 4-methoxyglucobrassicin, glucobrassicin,
272 and neoglucobrassicin in pakchoi [44]. The antioxidant activity and the total
273 phenolic content of Orthosiphon stamineus under an open environment were
274 higher than shade-grown conditions [45]. Total phenolic content in the
275 leaves of Ipomoea batatas was higher under 16 h than 8 h at light intensity
276 of 150 µmol·m-2·s-1 [46]. Antioxidant capacity and total phenolic content in
277 lettuce continuously increased with raising photoperiods under 150
278 µmol.m−2.s−1 condition. Specifically, the phenolic content in lettuce was
279 highest at the 24 h under 150 µmol.m−2.s−1, and was 5.3-fold higher than
280 under a 12 h period treatment [47]. Likewise, in the results of this
281 experiment, the total glucosinolate concentration increased with increasing
 18 Agriculture 2021, 11, x FOR PEER REVIEW 9 of 14
19

282 photoperiod and decreasing light intensity. This indicates that the long
283 photoperiods had a more photomorphogenic effect than a photosynthetic
284 one. However, the total glucosinolate concentration was not significantly
285 different between 20 h- and 24 h-photoperiod. The results indicate that the
286 total glucosinolate concentration could increase with increasing
287 photoperiods under low light intensity. However, it might reach a saturation
288 point under low light intensity and long photoperiod. Expanding the
289 photoperiod in a weak light intensity condition has a slight compensatory
290 effect, because it can decrease the negative influences of the weak light
291 stress.

292 Table 2. The individual glucosinolate concentration of watercress under different

293 lighting treatments of 12 h-266 µmol.m−2.s−1, 16 h-200 µmol.m−2.s−1, 20 h-160
294 µmol.m−2.s−1, and 24 h-133 µmol.m−2.s−1.

The individual glucosinolate concentration in watercress

shoot Lighting
(mg.g-1 DW)z treatments
Nastur Metho Brassi Hirsu Siber
15.69b 0.58b 1.83ab 0.53 1.04b 12 h-266 µmol
18.92a 0.68ab 1.75ab 0.45 0.63b 16 h-200 µmol
18.81ab 0.94a 1.40b 0.59 3.36a 20 h-160 µmol
20.71a 0.79ab 2.21a 0.53 0.46b 24 h-133 µmol
* *** ** NS *** Significancey

295 z
Siber: Glucosiberin, Hirsu: Glucohirsutin, Brassi: Glucobrassicin, Metho: 4-
296 Methoxyglucobrassicin, Nastur: Gluconasturtiin.

297 y
Different letters show a significant difference within each treatment by
298 Tukey’s multiple range test at NSNot significant (p > 0.05), *p ≤ 0.05; **p <
299 0.01; and ***p < 0.001 (n = 3).
 20 Agriculture 2021, 11, x FOR PEER REVIEW 10 of 14
21

300
301 Figure 3. The glucosinolate concentration (A) and content (B) in watercress shoot
302 under different lighting treatments of 12 h-266 µmol .m−2.s−1, 16 h-200 µmol.m−2.s−1,
303 20 h-160 µmol.m−2.s−1, and 24 h-133 µmol.m−2.s−1. Different letters above bars show
304 significant differences at p ≤ 0.05, using Tukey’s multiple range test (n = 3).

305 4. Conclusion
306 The results indicated that a photoperiod of 20 h at 160 µmol .m−2.s−1
307 enhanced total glucosinolate content and plant biomass of watercress grown
308 in a plant factory. Further studies can investigate the influence of light
309 quality from LEDs on the productivity and bioactive compounds of
310 watercress grown in a plant factory. Moreover, the results also suggested
311 that longer photoperiod induction was a potential method for watercress
312 glucosinolate production. There is great promise to apply these results to
313 improve the quality of watercress plants and enhance the efficiency in the
314 watercress cultivation in a plant factory.

315 Conflicts of Interest: Authors declare no conflict of interest

316 Acknowledgments: This work was supported by Institute of Information &
317 communications Technology Planning & Evaluation (IITP) grant funded by the Korea
318 government (MSIT) (No.2020-0-01441, Artificial Intelligence Convergence Research
319 Center (Chungnam National University)).

320 References
321
322 1. Zeb, A. Phenolic profile and antioxidant potential of wild watercress (Nasturtium officinale L.).
323 Springerplus. 2015, 4.
324 2. Klimek-Szczykutowicz, M.; Szopa, A.; Ekiert, H. Chemical composition, traditional and
325 professional use in medicine, application in environmental protection, position in food and
 22 Agriculture 2021, 11, x FOR PEER REVIEW 11 of 14
23

326 cosmetics industries, and biotechnological studies of Nasturtium officinale (watercress) - a

327 review. Fitoterapia. 2018, 129, 283-292.
328 3. EFSA European Food Safety Authority (EFSA) [(accessed on 25 January 2020)]; Available
329 online: http://www.efsa.europa.eu/.
330 4. Noia, J.D. Defining powerhouse fruits and vegetables: a nutrient density approach. Prev
331 Chronic Dis. 2014, 11, 130390.
332 5. Suroowan, S.; Mahomoodally, M.F. A comparative ethnopharmacological analysis of traditional
333 medicine used against respiratory tract diseases in Mauritius. J Ethnopharmacol. 2016, 177,
334 61-80.
335 6. Zafar, R.; Zahoor, M.; Shah, A.; Majid, F. Determination of antioxidants and antibacterial
336 activities, total phenolic, polyphenol and pigment contents in Nasturtium officinale. Pharmacol.
337 2017, 1, 11-18.
338 7. Klimek-Szczykutowicz, M.; Dziurka, M.; Blazevic, I.; Dulovic, A.; Granica, S.; Korona-Glowniak,
339 I.; Ekiert, H.; Szopa, A. Phytochemical and biological activity studies on Nasturtium officinale
340 (watercress) microshoot cultures grown in rita(r) temporary immersion systems. Molecules.
341 2020, 25.
342 8. Al-Kodmany, K. The vertical farm: A review of developments and implications for the vertical
343 city. Buildings. 2018, 8.
344 9. Ali, S.; Liu, Y.; Ishaq, M.; Shah, T.; Abdullah; Ilyas, A.; Din, I.U. Climate change and its impact
345 on the yield of major food crops: Evidence from pakistan Foods. 2017, 6.
346 10. Zhao, C., et al. Temperature increase reduces global yields of major crops in four independent
347 estimates. P Natl Acad Sci USA. 2017, 114, 9326-9331.
348 11. Kozai, T. Resource use efficiency of closed plant production system with artificial light:
349 Concept, estimation and application to plant factory. Proc. Jpn. Acad. Ser. 2013, 89, 447–461.
350 12. Kozai, T. Sustainable plant factory: Closed plant production systems with artificial light for high
351 resource use efficiencies and quality produce. Acta Hortic. 2013, 1004, 27–40.
352 13. Zakurin, A.O.; Shchennikova, A.V.; Kamionskaya, A.M. Artificial-light culture in protected
353 ground plant growing: Photosynthesis, photomorphogenesis, and prospects of LED application.
354 Russ J Plant Physl. 2020, 67, 413-424.
355 14. Liu, Y.; Ren, X.X.; Jeong, B.R. Supplementary Light source affects growth, metabolism, and
356 physiology of Adenophora triphylla (thunb.) A.DC. seedlings. Biomed Res Int. 2019, 2019.
357 15. Mao, H.P.; Hang, T.; Zhang, X.D.; Lu, N. Both multi-segment light intensity and extended
 24 Agriculture 2021, 11, x FOR PEER REVIEW 12 of 14
25

358 photoperiod lighting strategies, with the same daily light integral, promoted lactuca sativa l.
359 growth and photosynthesis. Agronomy. 2019, 9.
360 16. Kitaya, Y.; Niu, G.; Kozai, T.; Ohashi, M. Photosynthetic photon flux, photoperiod, and CO 2
361 concentration affect growth and morphology of lettuce plug transplants. HortScience 1998,
362 33, 988–991.
363 17. Vlahos, J.; Heuvelink, E.; Martakis, G. A growth analysis study of three Achimenes cultivars
364 grown under three light regimes. Sci. Hortic. 1991, 46, 275–282.
365 18. Zha, L.Y.; Liu, W.K. Effects of light quality, light intensity, and photoperiod on growth and yield
366 of cherry radish grown under red plus blue LEDs. Hortic Environ Biote. 2018, 59, 511-518.
367 19. Lu, N.; Bernardo, E.L.; Tippayadarapanich, C.; Takagaki, M.; Kagawa, N.; Yamori, W. Growth
368 and accumulation of secondary metabolites in perilla as affected by photosynthetic photon flux
369 density and electrical conductivity of the nutrient solution. Front Plant Sci. 2017, 8.
370 20. Kim, Y.J.; Kim, H.M.; Kim, H.M.; Jeong, B.R.; Lee, H.J.; Kim, H.J.; Hwang, S.J. Ice plant growth
371 and phytochemical concentrations are affected by light quality and intensity of monochromatic
372 light-emitting diodes. Hortic Environ Biote. 2018, 59, 529-536.
373 21. Fan, X.X.; Xu, Z.G.; Liu, X.Y.; Tang, C.M.; Wang, L.W.; Han, X.L. Effects of light intensity on the
374 growth and leaf development of young tomato plants grown under a combination of red and
375 blue light. Sci Hortic. 2013, 153, 50-55.
376 22. Kang, J.H.; KrishnaKumar, S.; Atulba, S.L.S.; Jeong, B.R.; Hwang, S.J. Light intensity and
377 photoperiod influence the growth and development of hydroponically grown leaf lettuce in a
378 closed-type plant factory system. Hortic Environ Biote. 2013, 54, 501-509.
379 23. Yin, Y.H.; Yu, C.J.; Yu, L.; Zhao, J.S.; Sun, C.J.; Ma, Y.B.; Zhou, G.K. The influence of light
380 intensity and photoperiod on duckweed biomass and starch accumulation for bioethanol
381 production. Bioresource Technol. 2015, 187, 84-90.
382 24. Rehman, M.; Ullah, S.; Bao, Y.; Wang, B.; Peng, D.; Liu, L. Light-emitting diodes: Whether an
383 efficient source of light for indoor plants? Environ Sci Pollut Res Int. 2017, 24, 24743–24752.
384 25. Taulavuori, E.; Taulavuori, K.; Holopainen, J.K.; Julkunen-Tiitto, R.; Acar, C.; Dincer, I. Targeted
385 use of LEDs in improvement of production efficiency through phytochemical enrichment. J Sci
386 Food Agr. 2017, 97, 5059-5064.
387 26. Engelen-Eigles, G.; Holden, G.; Cohen, J.D.; Gardner, G. The effect of temperature, photoperiod,
388 and light quality on gluconasturtiin concentration in watercress (Nasturtium officinale R. Br.). J
389 Agr Food Chem. 2006, 54, 328-334.
 26 Agriculture 2021, 11, x FOR PEER REVIEW 13 of 14
27

390 27. Choi, J.Y.; Kim, S.J.; Bok, K.J.; Lee, K.Y.; Park, J.S. Effect of different nutrient solution and light
391 quality on growth and glucosinolate contents of watercress in hydroponics. J Bio-Environ
392 Control. 2018, 27, 371-380.
393 28. Lam, V.P.; Choi, J.; Kim, S.; Park, J.; Hernandez, R. Optimizing plant spacing and harvest time
394 for yield and glucosinolate accumulation in watercress (Nasturtium officinale L.) grown in a
395 hydroponic system. Hortic Sci Technol. 2019, 37, 733-743.
396 29. Cuong, D.M.; Park, C.H.; Bong, S.J.; Kim, N.S.; Kim, J.K.; Park, S.U. Enhancement of
397 glucosinolate production in watercress (Nasturtium officinale) hairy roots by overexpressing
398 cabbage transcription factors. J Agr Food Chem. 2019, 67, 4860-4867.
399 30. ISO norm. Rapeseed-Determination of glucosinolate content. Part 1: Method using high-
400 performance liquid chromatography. ISO 9167-1. 1992, 1-9.
401 31. Li, Q.; Kubota, C. Effects of supplemental light quality on growth and phytochemicals of baby
402 leaf lettuce. Environ Exp Bot. 2009, 67, 59-64.
403 32. Solovchenko, A.E.; Khozin-Goldberg, I.; Didi-Cohen, S.; Cohen, Z.; Merzlyak, M.N. Effects of
404 light intensity and nitrogen starvation on growth, total fatty acids and arachidonic acid in the
405 green microalga Parietochloris incisa. J Appl Phycol. 2008, 20, 245-251.
406 33. Feng, L.Y., et al. The influence of light intensity and leaf movement on photosynthesis
407 characteristics and carbon balance of soybean. Front Plant Sci. 2019, 9.
408 34. Gommers, C.M.M.; Visser, E.J.W.; Onge, K.R.S.; Voesenek, L.A.C.J.; Pierik, R. Shade tolerance:
409 when growing tall is not an option. Trends Plant Sci. 2013, 18, 65-71.
410 35. Yang, F.; Huang, S.; Gao, R.C.; Liu, W.G.; Yong, T.W.; Wang, X.C.; Wu, X.L.; Yang, W.Y. Growth of
411 soybean seedlings in relay strip intercropping systems in relation to light quantity and red:far-
412 red ratio. Field Crop Res. 2014, 155, 245-253.
413 36. Schumann, T.; Paul, S.; Melzer, M.; Dormann, P.; Jahns, P. Plant growth under natural light
414 conditions provides highly flexible short-term acclimation properties toward high light stress.
415 Front Plant Sci. 2017, 8.
416 37. Austin, J.; Jeon, Y.A.; Cha, M.-K.; Park, S.; Cho, Y.-Y. Effects of photoperiod, light intensity and
417 electrical conductivity on the growth and yield of quinoa (Chenopodium quinoa Willd.) in a
418 closedtype plant factory system. Hortic Sci Technol. 2016, 34, 405-413.
419 38. Gong, W.Z., et al. Transcriptome analysis of shade-induced inhibition on leaf size in relay
420 intercropped soybean. Plos One. 2014, 9.
421 39. Shafiq, I., et al. Crop photosynthetic response to light quality and light intensity. J Integr Agr.
 28 Agriculture 2021, 11, x FOR PEER REVIEW 14 of 14
29

422 2021, 20, 4-23.

423 40. Cen, Y.P.; Sage, R.F. The regulation of rubisco activity in response to variation in temperature
424 and atmospheric CO2 partial pressure in sweet potato. Plant Physiol. 2005, 139, 979-990.
425 41. Su, B.Y.; Song, Y.X.; Song, C.; Cui, L.; Yong, T.W.; Yang, W.Y. Growth and photosynthetic
426 responses of soybean seedlings to maize shading in relay intercropping system in Southwest
427 China. Photosynthetica. 2014, 52, 332-340.
428 42. Zhu, H.F., et al. Effects of low light on photosynthetic properties, antioxidant enzyme activity,
429 and anthocyanin accumulation in purple pak-choi (Brassica campestris ssp Chinensis Makino).
430 Plos One. 2017, 12.
431 43. Huseby, S.; Koprivova, A.; Lee, B.R.; Saha, S.; Mithen, R.; Wold, A.B.; Bengtsson, G.B.; Kopriva,
432 S. Diurnal and light regulation of sulphur assimilation and glucosinolate biosynthesis in
433 Arabidopsis. J Exp Bot. 2013, 64, 1039-1048.
434 44. Yang, J.; Zhu, Z.J.; Gerendas, J. Interactive effects of phosphorus supply and light intensity on
435 glucosinolates in pakchoi (Brassica campestris L. ssp chinensis var. communis). Plant Soil.
436 2009, 323, 323-333.
437 45. Farhan, M.; Razak, S.A.; Pin, K.Y.; Chuah, A.L. Antioxidant activity and phenolic content of
438 different parts of Orthosiphon stamineus grown under different light intensities. J Trop for Sci.
439 2012, 24, 173-177.
440 46. Carvalho, I.S.; Cavaco, T.; Carvalho, L.M.; Duque, P. Effect of photoperiod on flavonoid pathway
441 activity in sweet potato (Ipomoea batatas (L.) Lam.) leaves. Food Chem. 2010, 118, 384-390.
442 47. Cho, J.Y.; Yoo, K.S.; Kim, J.; Choi, B.J.; Oh, W. Growth and bioactive compounds of lettuce as
443 affected by light intensity and photoperiod in a plant factory using external electrode
444 fluorescent lamps. Hortic Sci Technol. 2020, 38, 645-659.

445
446