Research Article

Received: 15 October 2012 Revised: 29 March 2013 Accepted article published: 12 April 2013 Published online in Wiley Online Library: 15 May 2013

(wileyonlinelibrary.com) DOI 10.1002/jsfa.6173

LED illumination affects bioactive compounds

in romaine baby leaf lettuce
Giedrė Samuolienė,∗ Aušra Brazaitytė, Ramūnas Sirtautas, Akvilė Viršilė,
Jurga Sakalauskaitė, Sandra Sakalauskienė and Pavelas Duchovskis
Abstract
BACKGROUND: The effect of light quality on phytochemicals in romaine baby leaf lettuce ‘Thumper’ was investigated in (I) a
closed environment and (II, III) a greenhouse (16 h, 21/17 ◦ C): (I) basal (638, 455, 660, 735 nm) LEDs supplemented with UV (380
nm), green (510 nm), yellow (595 nm) or orange (622 nm) LEDs (PPFD of ∼175 µmol m−2 s−1 ); (II) high-pressure sodium (HPS)
lamps (90 µmol m−2 s−1 ) supplemented with blue (455, 470nm) or green (505, 530nm) LEDs (30 µmol m−2 s−1 ); (III) at 3 days
before harvesting, HPS lamps (90 µmol m−2 s−1 ) supplemented with red (638 nm) LEDs (210 µmol m−2 s−1 ).
RESULTS: (I) Supplemental UV or orange light enhanced phenolic compounds, supplemental UV or green light enhanced
α-carotene, and supplemental green light enhanced anthocyanins. All supplemental LED colours had a negative effect on
tocopherol and ascorbic acid levels. (II) HPS lighting supplemented with different LEDs was not efficient, since the increase
in some compounds did not compensate the decrease in major tested phytochemicals. (III) Short-term irradiation with
supplemental 638 nm LEDs before harvesting in the greenhouse did not have a significant effect on phytochemical contents,
apart from enhancing tocopherols.
CONCLUSION: Wavelength control using LED technology affects the production of secondary metabolites, as the metabolism
of many nutrients is light-dependent. The narrow-bandwidth supplemental light effects were diminished by broader-spectrum
HPS light or natural daylight in the greenhouse.


c 2013 Society of Chemical Industry

Keywords: anthocyanins; ascorbate; carotenoids; DPPH; phenols; tocopherols

INTRODUCTION different technology compared with metal halide or high-pressure

Light modulates most biological processes and several metabolic sodium (HPS) lamps and enables one to select a particular
pathways, affecting the metabolism of bioactive compounds spectral composition and produce very high light levels with
and generating physiological responses.1 Photosynthetic and low radiant heat output.7 These LED-based technology properties
photomorphogenetic light receptors are responsible for the satisfy plant requirements for growth and development and thus
photophysiological responses incited by changes in light spectral are attractive for horticultural applications.8 On the other hand,
composition and also regulate light-sensitive molecular and LEDs emit a narrow-band spectrum, so plants must acclimatise
metabolic pathways. It is widely reported that red light is their photosynthetic functioning to such narrow-band lighting.9
The hypothesis was raised that lighting conditions might evoke
absorbed by chlorophylls and anthocyanins, and changes in red
photo-oxidative changes in plants, leading to increased content
and far-red light cause reversion of phytochromes. UV light is
and activity of phytochemical compounds. Seeking to evaluate the
absorbed by flavonoids, phenolics and anthocyanins.1,2 Blue light
LED light effect in different lighting environments and to establish
is absorbed by carotenoids, cryptochromes and phototropins,3
the effect on phytochemicals in romaine baby leaf lettuce (Lactuca
while green light responses can be both cryptochrome-dependent
sativa L.), experiments were performed in growth chambers under
and cryptochrome-independent.4 Moreover, light can act as a
controlled environmental conditions and in a greenhouse.
photostressor for plants, evoking antioxidant defence system
responses and thus affecting levels of protective phytochemical
compounds. Bioactive compounds can be divided into at least two EXPERIMENTAL
categories: those that directly absorb light stimulus (phenolics, Growing conditions and lighting systems
carotenoids) and those that are indirectly affected through Romaine green baby leaf lettuce (L. sativa L.) ‘Thumper’ (Nunhems
metabolic pathways (ascorbic acid, tocopherols). While it is widely Netherlands BV, Nunhem, Netherlands), which has long cylindrical
understood that light intensity can positively affect phytochemical
accumulation, the effects of light quality are more complex and
∗
often reported with mixed results.5 Correspondence to: Giedrė Samuolienė, Institute of Horticulture, Lithuanian
Plants usually receive a wide sunlight spectrum, emitted in Research Centre for Agriculture and Forestry, Kaunas Str. 30, LT-54333, Babtai,
Kaunas, Lithuania. E-mail: [email protected]
the visible region, or artificial lighting, where the spectrum
depends on the type of lamp chosen.6 Solid state lighting Institute of Horticulture, Lithuanian Research Centre for Agriculture and
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using light-emitting diodes (LEDs) represents a fundamentally Forestry, Kaunas Str. 30, LT-54333, Babtai, Kaunas, Lithuania

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c 2013 Society of Chemical Industry
Lettuce phytochemicals depend on light quality www.soci.org

leaves with serrated leaf margins, was grown to harvest time (22 The PPFD generated by each lighting unit was measured at the
days) in (I) a phytotron and (II, III) a greenhouse. Lettuces were top of the plant using a photometer/radiometer (RF-100, Sonopan,
sown and grown in peat substrate (pH 5–6; N 70, P 40, K 160, Poland). The PPFD was periodically adjusted to maintain the same
Ca 250, Mg 50 mg L−1 ) in 28-cell plastic plug trays (cell volume light level during the growth period at the top of the plant.
120 mL) at five seeds per cell. After germination, seedlings were
thinned out to leave three plants per cell. Trays were equally Sampling
distributed to act as three experimental replications for each At 22 days after seeding, lettuces from the central part of
lighting treatment. Day/night temperatures were 20 ± 2/16 ± 2 the illuminated area were harvested, excluding marginal plants
◦
C and relative air humidity was 50–60% in all treatments. Plants because of possible side effects. Conjugated biological samples
were watered when needed and fertilised with 2 g L−1 ammonium of the green matter of five randomly selected plants from
nitrate solution once a week. each treatment were used for analysis. Three replications were
(I) Lighting experiments were performed in growth chambers performed for each phytochemical measurement. All data are
under controlled environmental conditions, with natural expressed on a fresh weight (FW) basis.
illumination eliminated. The experiments started from sowing
time and lasted for 22 days until harvesting. Originally designed7 Determination of total phenolic compounds
LED-based lighting units consisting of commercially available The total content of phenolic compounds was determined
wavelengths were used: basal red (LXHL-LR3C, λ = 638 ± 23 nm in lettuce leaf 800 g L−1 methanol (POCH, Gliwice, Poland)
and L670-66-60, λ = 670 ± 23 nm; Philips Lumileds Lighting extracts (1:10 w/v) using a calorimetric Folin–Ciocalteu method.11
Company, San Jose, CA, USA), blue (Luxeon LXHL-LRC, λ = 455 ± 23 The absorbance at 765 nm was measured with a Genesys 6
nm; Philips Lumileds Lighting Company) and far-red (L735-05-AU, spectrophotometer (Thermo Spectronic, Rochester, NY, USA)
λ = 735 ± 25 nm; Epitex, Kyoto, Japan) and supplemental UV against water as a blank. Total phenolics were quantified by a
(NCCU001E, λ = 380 ± 15 nm; Nichia, Tokyo, Japan), green (LXHL- calibration method using gallic acid as a standard.
MM1D, λ = 510 ± 37 nm; Philips Lumileds Lighting Company),
yellow (LXHL-MLAC, λ = 595 ± 17 nm; Philips Lumileds Lighting 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free
Company) and orange (LXHL-MHAC, λ = 622 ± 20nm; Philips radical-scavenging activity
Lumileds Lighting Company) LEDs. A combination of blue 455 The antioxidant activity of lettuce leaf 800 g L−1 methanol
nm (8 µmol m−2 s−1 ), red 638 nm (150 µmol m−2 s−1 ), red 670 extracts (1:10 w/v) was evaluated spectrophotometrically as the
nm (12 µmol m−2 s−1 ) and far-red 735 nm (4 µmol m−2 s−1 ) light DPPH (Sigma-Aldrich, Munich, Germany) free radical (DPPH• )-
was used as the basal illumination spectrum. Other treatments scavenging ability (µmol g−1 ).11 A Genesys 6 spectrophotometer
were supplemented with UV 380 nm (4 µmol m−2 s−1 ; basal + UV), (Thermo Spectronic) was used for the analysis. The absorbance at
green 520 nm (12 µmol m−2 s−1 ; basal + G), yellow 595 nm (10 515 nm was measured after 16 min.
µmol m−2 s−1 ; basal + Y) or orange 622 nm (28 µmol m−2 s−1 ;
basal + O) LED wavelengths. In all treatments a photoperiod of 16 Determination of ascorbic acid
h was used and a total photosynthetic photon flux density (PPFD) Ascorbic acid (Penta, Prague, Czech republic) content was
of about 175 µmol m−2 s−1 was maintained by adjusting the flux determined using a spectrophotometric method12 based on
of red 638 nm LEDs. methyl viologen (Aldrich, Taufkirchen, Germany) reduction. A
(II, III) Plants were grown in a VD-Block-type greenhouse (Danish Genesys 6 spectrophotometer (Thermo Spectronic) was used for
Greenhouse Supply A/S, Tommerup, Denmark) in Lithuania the analysis. The coloured radical ion was measured at 600 nm and
(latitude 55◦ N) in November. Natural illumination (daily average ascorbic acid contents were quantified using a calibration method.
of 20–80 µmol m−2 s−1 ) was supplemented with standard HPS
lamps (SON-T Agro, 400 W; Philips, Somerset, NJ, USA) (PPFD of Determination of total anthocyanins
∼90 µmol m−2 s−1 ) and LED (II and III) lighting with a photoperiod The total amount of anthocyanins was determined using a
of 16 h. spectrophotometric pH-differential method.13 The absorption
(II) Four sets of solid state lamps for supplemental lighting in the values at 420, 520 and 700 nm were measured. Anthocyanins
greenhouse were designed using high-power blue 455 ± 23 nm were expressed as mg cyanidin 3-glucoside equivalent g−1 FW,
(LXHL-LR3C) or 470 ± 25 nm (LXHL-LB3C) or green 505 ± 30 nm using a molar extinction coefficient of 25 740 L mol−1 cm−1 and a
(LXHL-LE3C) or 530 ± 38 nm (LXHL-LM3C) AlInGaN LEDs (Luxeon molecular weight of 485 g mol−1 .
III series, 3 W; Philips Lumileds Lighting Company) that generated
a PPFD of 30 µmol m−2 s−1 . Plants were illuminated with LEDs and Determination of tocopherols
HPS lamps from sowing time until harvesting (22 days). Reference The contents of α- and γ -tocopherol (Supelco, Bellefonte, PA,
plants were grown under HPS lighting (PPFD increased to ∼115 USA) in lettuce leaf/hexane (Merck, Darmstadt, Germany) extracts
µmol m−2 s−1 ). (1:10 w/v) were determined using a high-performance liquid chro-
(III) Lettuces were grown under daylight with supplemental matography (HPLC) method. Samples were centrifuged (349 × g,
HPS lighting (90 µmol m−2 s−1 , 16 h) from sowing time. At 3 days 5 min) and filtered through a 0.45 µm polytetrafluoroethylene
before harvest (vegetative development stage), supplemental (PTFE) syringe filter (VWR International, Radnor, PA, USA). An HPLC
red LED illumination (16 h photoperiod) was added. The solid 10A system equipped with an RF-10A fluorescence detector (Shi-
state illuminator10 contained red AlGaInP LEDs (Luxeon III Star madzu, Kyoto, Japan) and a Pinacle II silica column (5 µm particle
model LXHL-LD3C, Philips Lumileds Lighting Company) with a size, 150 mm × 4.6 mm; Restec, Bellefonte, PA, USA) were used for
peak wavelength of 638 ± 23 nm. The PPFD generated by the the analysis. The mobile phase was 5 g L−1 isopropanol (Merck) in
LEDs was 210 µmol m−2 s−1 . Reference plants were grown under hexane at a flow rate of 1 mL min−1 . The peak was detected using
HPS lighting (PPFD increased to ∼300 µmol m−2 s−1 ) during the an excitation wavelength of 295 nm and an emission wavelength
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3 days until harvesting. of 330 nm.

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 www.soci.org G Samuolienė et al.

Determination of carotenoids induced significantly higher α-tocopherol (41.2% higher than

The contents of α- and β-carotene (ChromaDex, Irvine, CA, USA) HPS lighting) and γ -tocopherol (13.2% higher than HPS lighting)
in lettuce leaf 800 g L−1 ice-cold acetone (Merck, Darmstadt, contents in baby leaf lettuce, whereas supplemental blue 470
Germany) extracts (1:10 w/v) were determined using an HPLC nm light resulted in higher ascorbic acid concentration (53.8%
method. Samples were centrifuged (349 × g, 5 min) and filtered higher than HPS lighting). Supplemental cyan green 505 nm
through a 0.45 µm PTFE syringe filter (VWR International). An HPLC light resulted in significantly higher α-carotene content (33.3%
10A system equipped with an SPD-M 10A diode array detector higher than HPS lighting). Supplemental green 530 nm light
(Shimadzu) and a Zorbax Extended-C18 column (5 µm particle size, significantly stimulated total anthocyanin accumulation (38.9%
150 mm × 4.6 mm; Agilent Technologies, Waldbronn, Germany) higher than HPS lighting) and DPPH• -scavenging ability (9.6%
were used for the analysis. The mobile phase comprised 800 g L−1 higher than HPS lighting). A marked negative effect of all
methanol (Merck) and ethyl acetate (Sigma-Aldrich) at a flow rate tested supplemental LED light wavelengths on total phenolic
of 1 mL min−1 . The peak was detected at 440 nm. compound and β-carotene contents was observed. According to
the correlation analysis results (see Table 4), there were significant
Statistical analysis strong positive correlations between DPPH• -scavenging ability
and tocopherols in baby leaf lettuce plants grown under
Data were subjected to one-way analysis of variance (ANOVA)
supplemental blue (HPS + 470 nm) and green (HPS + 530 nm)
at the confidence level P ≤ 0.05. Values are presented as
LED light. In lettuce grown under supplemental blue 455 nm light,
mean ± standard deviation. MS Excel Version 7.0 was used for
significant strong positive correlations were determined between
data processing.
DPPH• -scavenging ability and carotenes and between phenolic
compounds and anthocyanins.
RESULTS
Experiment (I) Experiment (III)
The effect of LED light quality on phytochemicals in lettuce Short-term high-PPFD red LED lighting (HPS + 638 nm) resulted
grown in the controlled environment of growth chambers was in 13.7-fold higher α-tocopherol and 5.2% higher γ -tocopherol
variable (Table 1). Although no significant UV, green, yellow or contents as compared with HPS lighting alone (Table 3). On the
orange light effect on DPPH• -scavenging ability was determined other hand, a significant decrease in β-carotene content (42.5%
in comparison with basal LED illumination, different supplemental lower than HPS lighting) was observed. There was no significant
light wavelengths both positively and negatively affected levels effect on total phenol, total anthocyanin, ascorbic acid and α-
of phytochemical compounds. Supplemental UV (basal + UV) and carotene accumulation or DPPH• -scavenging ability. However,
orange (basal + O) light resulted in significantly higher contents of according to the correlation analysis results (Table 4), there was a
phenolic compounds in baby leaf lettuce (61.1 and 83.3% higher significant strong positive correlation between DPPH• -scavenging
than basal illumination respectively). The anthocyanin content was ability and total phenolic compounds.
22.6% higher in lettuce grown under supplemental green light
(basal + G) as compared with basal illumination, while α-carotene
contents were also found to be markedly higher after exposure of DISCUSSION
lettuce to UV (6.1 times higher than basal illumination) and green LED lighting allows the creation of efficient lighting facilities
(1.6 times higher than basal illumination) light. Supplemental LED with particular band waves and/or PPFD levels that match plant
lighting had a significant negative effect on ascorbic acid and α- physiological requirements and do not waste energy on other
and γ -tocopherol concentrations in lettuce leaves as compared unprofitable spectral parts such as green or yellow. It was shown
with basal illumination. Changes in β-carotene content were that red light alone in the lighting spectrum is sufficient for
statistically insignificant. lettuce growth and photosynthetic functioning; however, the
After different lighting exposures, strong positive correlations addition of blue14 – 16 and far-red17 light also has important effects
between antioxidant compounds were determined in most on growth parameters. Previously we have reported that the
cases (see Table 4). The exception was supplemental orange biometric characteristics, photosynthetic pigment content and
light (basal + O), which resulted in strong negative correlations photosynthesis intensity of lettuce can be optimised by adjusting
between anthocyanins and ascorbic acid, between anthocyanins
the light spectrum and flux density18 and that a combination of
and carotenes, between total phenols and anthocyanins and
basal blue, red and far-red LEDs matching the absorption spectrum
between DPPH• -scavenging ability and anthocyanins. Significant
of photosynthetic and photomorphogenetic light receptors3,19 is
strong positive correlations of DPPH• -scavenging ability with
suitable for lettuce cultivation. In this study we demonstrate
phenols and ascorbic acid were determined in lettuce
that basal blue, red and far-red LEDs are suitable for romaine
cultivated under basal + G illumination. Significant strong positive
baby leaf lettuce cultivation, since they lead to the accumulation
correlations were also determined between anthocyanins and
of appropriate levels of nutritionally important phytochemicals
carotenes in baby leaf lettuce cultivated under basal, basal + G
(Table 1). Other atypical light spectral components such as
and basal + Y illumination.
green and yellow were also proved to have significant effects
on lettuce growth.20,21 Our results demonstrate the marked
Experiment (II) effect of supplemental UV, green, yellow and orange lighting
Experiments in the greenhouse, where standard HPS lamps were on phytochemical contents. The addition of supplemental UV-
supplemented with blue (455, 470 nm) or green (505, 530 nm) A or orange light to the spectrum resulted in a significantly
LED components, revealed a very slight positive LED effect on higher concentration of total phenols. This is in agreement
the investigated phytochemical contents in comparison with with the literature, as phenolics are accumulated by a broad
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HPS lighting alone (Table 2). Supplemental blue 455 nm light range of plants following UV exposure.22 An increase in phenolic

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c 2013 Society of Chemical Industry J Sci Food Agric 2013; 93: 3286–3291
Lettuce phytochemicals depend on light quality www.soci.org

Table 1. Effect of LED light quality on phytochemicals in baby leaf lettuce grown in growth chambers

Phytochemical Basal Basal + UV Basal + G Basal + Y Basal + O

Phenols (mg g−1 ) 1.44 ± 0.05 2.32 ± 0.08∗ 1.52 ± 0.03 1.59 ± 0.16 2.64 ± 0.04∗
DPPH• (µmol g−1 ) 10.02 ± 0.19 9.44 ± 0.66 8.97 ± 0.93 9.27 ± 0.30 9.64 ± 0.19
Anthocyanins (mg g−1 ) 84.98 ± 5.97 75.19 ± 4.63 104.15 ± 7.55∗ 77.95 ± 3.23 51.92 ± 5.22∗
Ascorbic acid (mg g−1 ) 0.35 ± 0.03 0.30 ± 0.00∗ 0.33 ± 0.04 0.29 ± 0.01∗ 0.32 ± 0.01
α-Tocopherol (µg g−1 ) 0.86 ± 0.01 0.31 ± 0.00∗ 0.59 ± 0.01∗ 0.37 ± 0.01∗ 0.40 ± 0.01∗
γ -Tocopherol (µg g−1 ) 8.54 ± 0.16 7.30 ± 0.08∗ 8.05 ± 0.18∗ 8.12 ± 0.13∗ 7.33 ± 0.10∗
β-Carotene (µg g−1 ) 2.17 ± 0.31 1.86 ± 0.49 1.86 ± 0.43 2.78 ± 0.38 1.72 ± 0.51
α-Carotene (µg g−1 ) 0.22 ± 0.03 1.35 ± 0.06∗ 0.36 ± 0.02∗ 0.24 ± 0.12 0.24 ± 0.04

Basal, 455, 638, 670 and 735 nm; UV, 380 nm; G (green), 520 nm; Y (yellow), 595 nm; O (orange), 622 nm.
∗ Significantly (P ≤ 0.05) different from basal value.

Table 2. Effect of supplemental blue (455, 470 nm) and green (505, 530 nm) LED light on phytochemicals in baby leaf lettuce grown in a greenhouse
under HPS lighting

Phytochemical HPS HPS + 455 nm HPS + 470 nm HPS + 505 nm HPS + 530 nm

Phenols (mg g−1 ) 1.00 ± 0.04 0.61 ± 0.03∗ 0.58 ± 0.03∗ 0.87 ± 0.03∗ 0.71 ± 0.03∗
DPPH• (µmol g−1 ) 9.95 ± 0.09 9.71 ± 0.09∗ 8.32 ± 0.05∗ 9.97 ± 0.09 10.91 ± 0.08∗
Anthocyanins (mg g−1 ) 57.64 ± 7.66 53.17 ± 3.17 62.94 ± 2.94 54.17 ± 4.31 80.09 ± 3.05∗
Ascorbic acid (mg g−1 ) 0.26 ± 0.00 0.21 ± 0.01∗ 0.40 ± 0.02∗ 0.28 ± 0.02 0.26 ± 0.00
α-Tocopherol (µg g−1 ) 1.70 ± 0.07 2.4 ± 0.08∗ 1.61 ± 0.01 1.57 ± 0.02∗ 1.59 ± 0.01∗
γ -Tocopherol (µg g−1 ) 8.06 ± 0.24 9.12 ± 0.18∗ 6.54 ± 0.14∗ 7.89 ± 0.07 7.17 ± 0.13∗
β-Carotene (µg g−1 ) 0.28 ± 0.04 0.20 ± 0.01∗ 0.09 ± 0.01∗ 0.23 ± 0.03 0.22 ± 0.02∗
α-Carotene (µg g−1 ) 2.16 ± 0.06 1.24 ± 0.02∗ 0.67 ± 0.02∗ 2.88 ± 0.27∗ 1.19 ± 0.04∗
∗
Significantly (P ≤ 0.05) different from HPS value at same PPFD level.

combining the spectra of these conventional light sources with

Table 3. Effect of short-term high-PPFD red 638 nm LED lighting
applied 3 days before harvest on phytochemicals in baby leaf lettuce LED wavelengths, it is possible not only to optimise spectral quality
grown in a greenhouse under HPS lighting for various plants but also to create economically efficient lighting
systems. In order to enrich the spectrum of HPS lamps, which
Phytochemical HPS HPS + 638 nm emit more yellow and red light and less blue light,6 blue, cyan
Phenols (mg g−1 ) 0.63 ± 0.05 0.64 ± 0.03 and green LEDs were added to the lighting spectrum. Blue light,
DPPH• (µmol g−1 ) 10.13 ± 0.20 10.07 ± 0.31 applied in combination with red LEDs or fluorescent lamps in
Anthocyanins (mg g−1 ) 74.25 ± 4.23 78.13 ± 5.26 closed growth chambers, was reported to stimulate antioxidant
Ascorbic acid (mg g−1 ) 0.38 ± 0.03 0.33 ± 0.04 status in green vegetables by enhancing polyphenol,16 vitamin
α-Tocopherol (µg g−1 ) 0.79 ± 0.02 11.60 ± 0.08∗ C,26 carotenoid5,27 and anthocyanin5,15 contents. However, in the
γ -Tocopherol (µg g−1 ) 206.11 ± 3.21 216.93 ± 2.16∗ greenhouse environment, supplemental blue and green LEDs did
β-Carotene (µg g−1 ) 7.09 ± 0.92 4.08 ± 0.85∗ not have a versatile positive effect on phytochemical contents
α-Carotene (µg g−1 ) 0.72 ± 0.01 0.70 ± 0.02 (Table 1). It was reported that successful lighting strategies in
phytotrons did not necessarily produce the same results under
∗
Significantly (P ≤ 0.05) different from HPS value at same PPFD level. greenhouse conditions,28 especially when the variable effect of
daylight is also involved in the complex exposure. According to our
results, supplemental blue light had a negative effect on phenolic
concentration in lettuce was observed after UV-A LED exposure23 compound and carotene contents and DPPH• -scavenging ability;
as well as using UV-A fluorescent lamps or light filters.24,25 however, blue 455 nm light enhanced tocopherols, while blue 470
Moreover, according to our data, significantly higher carotene nm light had a positive effect on ascorbic acid. Cyan green 505 nm
but lower tocopherol contents were observed under basal + UV light had the least effect, while green 530 nm light resulted in higher
LED treatment (Table 1). Thus we hypothesise that compensatory antioxidant activity, possibly owing to the marked increase in leaf
interaction between antioxidants occurs in lettuce plants exposed anthocyanin concentration. Our previous results29 with different
to different light spectra. In general, other supplemental LED red, green and light green curly leaf lettuce varieties showed that
light colours (green, yellow, orange) had a negative effect on green 535 nm LEDs had a greater positive effect on ascorbic acid
tocopherol and ascorbic acid contents, though LED light spectra and tocopherol contents and DPPH• -scavenging capacity, while
are potent to enhance the contents of selected phytochemicals in cyan green 505 nm LEDs had a greater effect on total phenol
lettuce plants. and anthocyanin contents. Blue 455 and 470 nm light had the
LED technology to date is still too expensive to displace least expressed effect on phytochemical contents. Moreover, the
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fluorescent or HPS illumination in greenhouses. However, by metabolism of antioxidants in lettuce was dependent on multiple

J Sci Food Agric 2013; 93: 3286–3291

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 www.soci.org G Samuolienė et al.

Table 4. Correlation (r) between phytochemicals in lettuce leaves exposed to different light qualitites

Growth chambers Greenhouse

Correlation Basal Basal Basal Basal HPS HPS HPS HPS HPS
between Basal + UV +G +Y +O HPS + 455 nm + 470 nm + 505 nm + 530 nm + 638 nm

Phenols DPPH• 0.98 1.00 1.00∗ 0.66 0.94 0.98 0.98 0.83 0.98 0.77 1.00∗
Anthocyanins 1.00∗ 0.96 1.00 0.96 −0.96 0.99 1.00∗ 0.98 0.99 0.94 0.97
Ascorbic acid 0.90 0.99 1.00∗ 0.94 0.99 0.99 0.94 0.98 0.80 0.64 0.79
Tocopherols 0.75 0.91 0.77 0.68 0.90 0.93 0.94 0.81 0.81 0.71 0.84
Carotenes 1.00∗ 0.98 0.99 0.95 1.00 1.00 0.99 0.87 1.00∗ 0.97 0.89
DPPH• Anthocyanins 0.98 0.92 0.99 0.84 −0.82 0.95 0.97 0.91 1.00 0.94 0.98
Ascorbic acid 0.97 0.96 1.00∗ 0.87 0.89 0.95 0.86 0.91 0.90 0.99 0.81
Tocopherols 0.86 0.94 0.73 1.00∗ 0.99 0.85 0.99 1.00∗ 0.91 1.00∗ 0.87
Carotenes 0.97 1.00 0.98 0.86 0.98 1.00 1.00∗ 1.00 0.96 0.89 0.91
Anthocyanins Ascorbic acid 0.90 0.99 0.99 1.00∗ −0.99 1.00 0.96 1.00 0.87 0.87 0.92
Tocopherols 0.74 0.75 0.82 0.85 −0.75 0.97 0.92 0.90 0.87 0.91 0.95
Carotenes 1.00∗ 0.89 1.00∗ 1.00∗ −0.92 0.97 0.98 0.95 0.98 0.99 0.98
Ascorbic acid Tocopherols 0.96 0.82 0.72 0.88 0.84 0.97 0.78 0.90 1.00∗ 1.00 0.99
Carotenes 0.87 0.94 0.98 1.00∗ 0.97 0.97 0.89 0.95 0.76 0.80 0.98
Tocopherols Carotenes 0.71 0.97 0.85 0.88 0.95 0.90 0.97 0.99 0.77 0.86 1.00

Basal, 455, 638, 670 and 735 nm; UV, 380 nm; G (green), 520 nm; Y (yellow), 595 nm; O (orange), 622 nm; HPS, high-pressure sodium lighting.
∗ P ≤ 0.05.

factors such as variety, light quality and season. Our present results Niki and Noguchi36 state that individual antioxidant compounds
showed that the obtained increase in some compounds did not act in combination with other antioxidants, as interactions among
compensate the decrease in other tested phytochemical contents them can affect total antioxidant capacity, producing synergistic
in romaine baby leaf lettuce (Table 2). This effect might be related or antagonistic effects. In our study the DPPH• -scavenging activity
to the dosage of blue3 and green30 light, as a positive response of was highly correlated with the contents of tested phytochemicals
phytochemicals was achieved when these spectral components (Table 4). In agreement with other authors,37 the linear correlation
accounted for up to 50% of the total photon flux density. between phenolics and DPPH• -scavenging ability suggests that
Another idea to incorporate LED lighting for improving the presence of phenolic compounds largely accounts for the
the nutritional quality of green vegetables without requiring antioxidant capacity. Hashempour et al.38 found that antioxidant
investment to install LEDs in large greenhouse areas was short- capacity was highly correlated with total anthocyanin and total
term preharvest treatment. A high flux of red 640 nm LED light phenolic contents, whereas the linear correlation between total
applied for 3–7 days before harvesting in controlled environment antioxidant capacity and ascorbic acid was moderate. However,
chambers27 or greenhouses, possibly acting as a photostressor, according to the present results, different light spectral quality
alters both the concentrations of individual phytochemicals
evoked an antioxidant system response and enhanced the
and the correlative interactions between them, indicating that
contents of lutein and glucosinolate sinigrin in red leaf cabbage27
different light colours initiate different light perception and
and phenolic compounds, ascorbic acid and α-tocopehrol in
signalling mechanisms.
lettuce31 and other green vegetables.32 Our previous results33
obtained from experiments with different red, green and light
green leaf curly lettuce varieties irradiated with supplemental 638
nm LED light in a greenhouse resulted in increases in total phenolic
CONCLUSIONS
Lighting wavelength control using LED technology can affect
compound content and free radical-scavenging activity in red
the production of secondary metabolites, as the synthesis or
and light green leaf lettuce. However, green leaf lettuce showed
absorption of many nutrients is light-dependent. Our results
statistically unreliable changes in vitamin C, total anthocyanins
showed the following. (I) Blue, red and far-red LEDs (basal)
and free radical-scavenging activity. Similar data were obtained
are suitable for romaine baby leaf lettuce cultivation, leading
in the present study with romaine green leaf lettuce, where
to the accumulation of appropriate levels of phenols, ascorbic
short-term high-PPFD red LEDs had no significant effect on total acid, anthocyanins, carotenes and tocopherols. Supplemental UV
phenol, total anthocyanin and ascorbic acid accumulation or light (basal + UV) enhanced phenolic compounds and α-carotene,
DPPH• -scavenging activity (Table 3). It is known that the red light supplemental green light (basal + G) enhanced α-carotene
signalling pathway is related to phytochrome transformation1 and anthocyanins, and supplemental orange light (basal + O)
and there is evidence suggesting that phenolic compound34 enhanced phenolic compounds. However, all supplemental LED
and anthocyanin35 synthesis is regulated by phytochromes. Also, colours had a negative effect on tocopherol and ascorbic acid
lighting conditions might evoke photo-oxidative changes in levels. (II) HPS lighting supplemented with blue, cyan or green
plants that lead to changed contents and activity of secondary LEDs in the greenhouse was not efficient, as the increase in
metabolites even if they do not participate in reactions directly some compounds did not compensate the decrease in major
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receptive to the light stimulus. tested phytochemicals. Supplemental green 530 nm light had the

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most pronounced effect on lettuce leaf extract DPPH• -scavenging 18 Brazaitytė A, Ulinskaitė R, Duchovskis P, Samuolienė G, Šikšnianienė
ability, mainly owing to the increased anthocyanin content. (III) JB, Jankauskienė J, et al., Optimization of lighting spectrum for
Short-term irradiation with supplemental red 638 nm LEDs before photosynthetic system and productivity of lettuce by using light-
emitting diodes. Acta Hort 711:183–188 (2006).
harvesting in the greenhouse did not have a significant effect on 19 Franklin KA and Whitelam GC, Light signals, phytochromes and cross-
phytochemical contents, apart from enhancing tocopherols. talk with other environmental cues. J Exp Bot 55:271–276 (2004).
The results demonstrated that the levels of selected 20 Kim HH, Goins GD, Wheeler RM and Sager JC, Green-light
phytochemicals in lettuce plants may be purposefully altered supplementation for enhanced lettuce growth under red- and
by irradiating plants with optimised lighting spectra or creating blue-light-emitting diodes. HortScience 39:1617–1622 (2004).
21 Johkan M, Shoji K, Goto F, Hahida S and Yoshihara T, Effect of
mild photostress. However, the mechanisms of the changes in green light wavelength and intensity on photomorphogenesis and
phytochemical contents under different light quality are not well photosynthesis in Lactuca sativa. J Environ Exp Bot 75:128–133
known yet. Moreover, the narrow-bandwidth supplemental light (2012).
effects were diminished by broader-spectrum HPS light or natural 22 Jansen MAK, Hectors K, O’Brien NM, Guisez Y and Potters G, Plant
stress and human health: do human consumers benefit from UV-B
daylight in the greenhouse. acclimated crops? Plant Sci 175:449–458 (2008).
23 Lee NY, Lee MJ, Kim YK, Park JC, Park HK, Choi JS, et al., Effect of light
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ACKNOWLEDGEMENT Soc Appl Biol Chem 53:658–690 (2010).
This work was supported by the framework of the programme 24 Tsormpatsidis E, Henbest RGC, Battey NH and Hadley P, The influence
of ultraviolet radiation on growth, photosynthesis and phenolic
‘Horticulture: agro-biological basis and technologies’. levels of green and red lettuce: potential for exploiting effects of
ultraviolet light in a production system. Ann Appl Biol 156:357–366
(2010).
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