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Determination of the Effect of Red and Blue

Ratios of LED Light on Plant Photosynthesis

Conference Paper July 2014

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 Invited Paper

Impact of LED Irradiance on Plant Photosynthesis and Action

Spectrum of Plantlet
Most Tahera Naznin*and Mark G. Lefsrud
Bioresource Engineering Department, Macdonald Stewart Building, McGill University, 21111
Lakeshore, Ste-Anne-de-Bellevue, H9X 3V9, Quebec, Canada
*most.naznin@mail.mcgill.ca; Phone: 1-514-398-7680; Fax: 1-514-398-8387

Keywords: light-emitting diode (LED), plantlets, photosynthesis, action spectrum

Abstract
Light emitting diodes (LEDs) can be selected to target the wavelengths absorbed by plantlets, enabling the users to
customize the wavelengths of light required for maximum production. The primary purpose of this experiment was to
test the effect of different ratios of red to blue LEDs on tomato plantlets photosynthetic action spectrum. Four light
treatments including: red LED (100%) and three ratios of red (661 nm) to blue (449 nm) light (5:1, 10:1 and 19:1) at 60
umol m-2 s-1 for this study. The tomato plantlets cultured without blue light showed a three and half-fold decrease in
photosynthesis rate. The highest photosynthetic action spectrum was observed at 10:1 but was not significantly
difference from the 5:1 and the lowest action spectrum was observed at 100% red LED light. The tomato plantlets grown
without the blue light showed a single-fold increase in plantlet height but were not significantly different from the 10:1
red to blue LED light. This research will allow for improved selection of LED lighting for plant tissue culture.

1. Introduction
Plants require light for photosynthesis, proper growth and physiological development [1]. Plants are
additionally sensitive to the spectral composition of their source of light, and the specific impact on plant responses of
photosynthesis, photomorphogenesis and phototropism [2, 3]. Plant growth and development are impacted by light
intensity, light quality, duration, and photoperiod [4]. Fluorescent lamps are the main light source commonly used for in
vitro cultivation of plants. Fluorescent lamps have fixed emission spectra composed of many bands in the wavelength
range from 320 to 800 nm without the possibility of varying illumination parameters (spectrum and pulse
characteristics). With the development of high-power LEDs, a wide range of emission wavelengths have become
commercially available for applications in plant cultivation [5]. When comparing LEDs to conventional fluorescent
lamps and high intensity discharge (HID) lamps, LED illuminators have improved features including smaller mass and
volume, and longer life spans [6, 7]. LED-based illuminators provide an alternative to fluorescent and HID lamps as a
light source with wavelength spectrum that can be selected for plant cultivation [8].
Growth, morphology, and differentiation of in vitro plantlets are also impacted by light quality [9]. Light is
needed by plants to mediate many hormonal and morphological changes, and specific wavelengths have been shown to
benefit plants in different ways. For example, both blue light (~420-450 nm) and red light (660 nm) have been shown to
increase chlorophyll production and play an active role in photosynthesis [10]. Both wavelengths have been shown to
improve morphological and physiological growth of plants [11, 12].
Red light has been shown to significantly enhance stem elongation of Pelargonium plantlets in vitro while blue
light inhibited shoot length [13]. Jao and Fang studied in vitro growth of potato plantlets using LEDs which stimulated
plant growth [16, 17]. Aksenova et al. (14) found that in vitro potato plantlets produced longer stems and higher root to
shoot ratio with red light than with blue light. Nhut et al. [15] showed that the best growth of strawberry plantlets
cultured in vitro was observed under LED-based illumination with 70% red and 30% blue spectral components,
respectively, while the optimal total PFD was found to be 60 mol m -2 s-1.
Tomato is one of the most important Solanaceae crop grown throughout the world [18]. Tomato is one of the
most consumed crops in the world. It is grown in tropical, sub-tropical and temperate regions [19]. Several in vitro
investigations have been conducted on tomato based on its relationship with tobacco, and in account of its consequently

Optics and Photonics for Information Processing VIII, edited by Abdul A. S. Awwal,
Khan M. Iftekharuddin, Mohammad A. Matin, Proc. of SPIE Vol. 9216, 921602
2014 SPIE CCC code: 0277-786X/14/$18 doi: 10.1117/12.2061236

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 expected good workability [20]. Shoot apex, nodal segments and root segments have been successfully used for callus
induction and regeneration [21].
This study was conducted to explore the regeneration potential of tomato hypocotyls segments under LED light
and to examine whether the light source results in a significant impact on the growth of tomato plant tissue culture.

2. Material and Methods

2.1. Plant materials and culture conditions
Surface sterilization of tomato (Solanum lycopersicum), seeds was performed by soaking the seeds in a solution of 8%
clorox (sodium hypochlorite) for 10 minutes, followed by three times rinsing with sterilized distilled water. Seeds were
inoculated in test tubes containing MS [22] medium and were transferred to a dark room for germination. Hypocotyl
segments (1-2 cm) of 18-21 day old in vitro plants were excised under aseptic conditions. The excised explants were
cultured in MS medium supplemented with 30 g sucrose and 2 mg/l of BAP. The cultures were incubated under different
light treatments in a growth chamber (E15, Conviron, Winnipeg, Canada) (16/8 light/dark regime, 231 OC temperature,
605 umol m-2 s-1 PAR) for six weeks. Data was collected and evaluated.

2.2. Light treatments

Cultures of in vitro plantlets were illuminated using red and blue ratios. Four light treatments included: red LED
(100%) and three LED ratios of red (661 nm) to blue (449 nm) light (5:1, 10:1 and 19:1) were used in this study. The
photoperiod was at 16/8 h (day/night) and photosynthetic photon flux density (PPFD) was 60 5 molm2s1 PPFD in
the growth chamber (LI-193; LI-COR Inc, Lincoln, NE, USA).

2.3. Experimental design and statistical analysis

Plant photosynthesis action spectrum and shoots length were measured for each treatment after six weeks.
Whole plant photosynthetic measurements were made using the LI-6400 photosynthesis system (LI-COR Inc., Lincoln,
NE, USA) equipped with a 6400 -17 Whole Plant Arabidopsis chamber (LI-COR Inc.). The LI-6400 controlled relative
humidity (75% +/-3.5%), CO2 concentration (400 ppm +/- 10 ppm) and temperature (21C +/- 1C). Photosynthesis rate
was normalized with leaf area. Photosynthetic action spectrum was measured according to McCree et al. (1972) [23].
2.4. Statistical analysis: The statistical analysis was made using analysis of variance (ANOVA). Least significant
differences were evaluated by the Tukey- Kramer method (P < 0.05).

1.1.1.1.1. 3. Results and discussion

The tomato plantlets grown without the blue component showed a three and half-fold decrease in the measured
photosynthesis rate (Figure 1). The highest photosynthesis rate of the tomato plantlets was observed at the 10:1 red to
blue ratio of LED light. The photosynthesis rate of the tomato plantlets were 3.5, 2.9 and 2.2 times greater, respectively,
10:1, 5:1 and 19:1 red to blue ratio than 100% red LED light.

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 (mm CO2 m-2 s-1) 3

b
Photosynthesis

2 a

a,c

1 d

0
5:1
5:01 10:1
10:01 19:1
19:01 100
100
Red to blue ratio
Figure 1. Photosynthesis of tomato plantlets under different ratios of red to blue led light. Values are the means of 5 determinations (n
= 5). LSD (P < 0.05) was determined according to Tukey-Kramers multiple range tests.

The photosynthetic action spectrum of tomato plantlets showed a similar tendency to the photosynthetic rate.
The plantlets grown without the blue light showed a three-fold decrease in action spectrum (Figure 2). The highest action
spectrum was observed at 10:1 red to blue ratio of LED light. Action spectrum was 3.4, 2.8 and 2.1 times, respectively,
greater at 10:1, 5:1 and 19:1 red to blue ratio than 100% red LED light.

0.04
a,b
Action Spectrum
(mm CO2 m-2 s-1)

c,d
0.02
d

0
5:1
5:01 10:1
10:01 19:1
19:01 100
Red to blue ratio
Figure 2. Action spectrum of tomato plantlets under different ratios of red to blue led light. Values are the means of 5 determinations
(n = 5). LSD (P < 0.05) was determined according to Tukey-Kramers multiple range tests.
The tomato plantlets grown without the blue light showed a single-fold increase in plantlet height (Figure 3).
The highest plantlet height was observed at 100% red LED light. The height of the tomato plantlet was less by 1.1, 1.3
and 1.4 times, respectively, for 10:1, 5:1 and 19:1 red to blue ratio compared to the 100% red LED light.

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 16
Plant height (cm)

d
12 b, d
a
c
8

0
5:1
5:01 10:1
10:01 19:1
19:01 100
100
Red to blue ratio
Figure 3. Plant height of tomato plantlet s under different ratios of red to blue led light. Values are the means of 5 determinations (n =
5). LSD (P < 0.05) was determined according to Tukey-Kramers multiple range tests.

Our experiment shows that the blue light in the illumination spectrum inhibits plantlet extension but resulted
increases in both the photosynthesis rate and photosynthetic action spectrum (Figure 1, 2 and 3). This is similar to
reported observations by Nhut et al. [15] in strawberry, Kim et al. in Chrysanthemum [24], and Jao et al. in Zantedeschia
[25] grown in vitro. The complex influence of the simultaneous illumination in blue and red/far-red regions might be
interpreted by synergistic interactions between the photoreceptors of blue and red light (cryptochromes and
phytochromes, respectively) [5].

In conclusion, among the four treatments of light, 10:1 ratio of red to blue LED was found superior in plantlet
photosynthesis and photosynthetic action spectrum. This research will allow for improved selection of LEDs for plant
tissue culture.

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