Bioponics for lettuce production in a plant factory with

artificial lighting
W. Fang a and H. Chung
Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, Taiwan, R.O.C.

Abstract
The plant factory with artificial lighting (PFAL) has gained worldwide attention,
and the most popular culturing system is hydroponics, in which pure soluble
inorganics fertilizers are used. The manufacturing processes of these fertilizers are
not environment friendly. A new trend termed ‘bioponics’, using organic fertilizers,
was brought to our attention. This study aimed to use two organic fertilizers (BIO N
and BIO NK, Swiss Hydroponics Lab.) in a deep-flow-technique (DFT) system in a PFAL
producing ‘Frill-ice’, ‘Boston’ and ‘Frilly’ lettuce, with Yamazaki formula (inorganic
fertilizer) as the control group. The environmental settings of the PFAL were as
follows: photosynthetic photon flux density (PPFD) 200 μmol m-2 s-1, day/night
temperature 25/18°C; CO2 concentration 1200 ppm. Our results showed that the
control group had the highest fresh mass for ‘Frill-ice’ lettuce. Bioponics treatments
greatly reduced the nitrate concentration of all lettuces and increased the fresh mass
of ‘Frilly’ lettuce. BIO N was not suitable for ‘Boston’ lettuce. Further studies will be
conducted using BIO NK and ‘Frill-ice’ lettuce. Using organic fertilizer in PFAL looks
promising; however, whether or not the products of bioponics can be considered
organic is still controversial.

Keywords: bioponics, hydroponics, deep flow technique, organic

INTRODUCTION
Soilless culture is a widely used technique to grow plants without soil. It can use
organic or inorganic substitute materials such as bark or rockwool or non-solid materials.
Hydroponics is a subset of soilless culture using a circulated nutrient solution. It provides a
considerable degree of control of the elemental environment surrounding the roots.
Hydroponic cultivation systems have been developed to control soil-borne pathogens and
are especially powerful in element-deficit research. The ability to use organic fertilizers in
hydroponics has been studied as a method to grow crops in space habitats (Garland et al.,
1997). The results showed that direct use of organic fertilizer proved to be deleterious to
plant growth.
For environmental protection and food safety reasons, organic agriculture has
garnered lots of public attention. Many people consider that organic agriculture can also
provide more nutritious food with better taste. This is controversial. The National Organic
Standards Board (NOSB) is a Federal Advisory Board in the USA made up of 15 dedicated
public volunteers from across the organic community. In 2016, the NOSB announced a
definition for the term ‘bioponics’.
Bioponics is a contained and controlled growing system in which plants in growing
media derive nutrients from plant-based, animal-based and mineral natural substances that
are released by the biological activity of microorganisms. Aquaponics and organic
hydroponics fit the definition of bioponics. There are three major roles in aquaponics:
fish/prawn, plant and beneficial bacteria. The fish/prawn wastes and uneaten feed
contained ammonia, which dissolved in water becomes ammonium ions, which can be
nitrified by bacteria to become nitrate and nitrite ions. Both ammonium and nitrate ions are

a
E-mail: weifang@ntu.edu.tw

Acta Hortic. 1227. ISHS 2018. DOI 10.17660/ActaHortic.2018.1227.75 593

Proc. Int. Symp. on New Technologies for Environ. Control, Energy-Saving and
Crop Production in Greenhouse and Plant Factory - GreenSys2017
Eds.: Qichang Yang and Tao Li
nutrients for the plants. Traditional hydroponics use inorganic fertilizers in the circulated
nutrient solution; organic hydroponics replace inorganic fertilizer with organic fertilizers,
which may be a combination of extracts from agriculture waste and/or plants/animals.
Shinohara et al. (2011) compared the production of hydroponically grown tomato
using organic and inorganic fertilizers. Their results appeared to show that roots grown with
inorganic fertilizer had no root hairs, and large numbers of root hairs developed on the roots
of tomato grown with fish-based soluble fertilizer. The hairs were also covered with biofilms
Organic hydroponics can also lead to induced systemic resistance (ISR) in lettuce against
airborne Botrytis cinerea, which causes gray mold (Chinta et al., 2015). It can be stated that
organic fertilizers can enhance/induce the growth of root hairs, thus providing better
absorption of nutrients and fewer airborne bacterium-induced diseases, leading to better
growth and yields.
The production of powdered inorganic fertilizers from minerals requires lots of energy
and creates lots of pollutants. The shipping and transporting of inorganic fertilizers from
manufacturing sites to the sites of plant growth also require lots of energy; thus, they are not
environment friendly compare with locally produced organic fertilizer. Most plant factories
with artificial lighting (PFALs) uses multilayer shelves equipped with hydroponic systems
such as NFT (nutrient film technique), DFT (deep flow technique) or E&F (ebb and flood). In
such systems, inorganic fertilizers are used. Recently, organic fertilizers have been tested,
which also brought lots of attention. The purposes of this study were to test three cultivars
of lettuce using two types of commercially available organic fertilizer in PFAL. To be used in
a confined environment such as PFAL, any material with a bad smell is not allowed, thus
making organic fertilizers extracted from plant waste a better choice.

MATERIALS AND METHODS

Culture conditions
Three cultivars of lettuce (Lactuca sativa L.), ‘Frill-ice’, ‘Boston’, and ‘Frilly’, were grown
in the PFAL of National Taiwan University. All seeds were locally available. Table 1 shows the
treatment code of three stages: the pre-treatment stage, seedling stage and mature stage.
The purpose for the pre-treatment stage is to culture the water, to allow the growth of
nitrifying bacteria in order to reduce the concentration of ammonium and increase the
concentration of nitrate. It required 21 days before sowing (DBS).

Table 1. Treatments and codes in stages of lettuce production with organic fertilizer. CK,
Control; DBS, days before sowing; DAS, days after sowing. Treatment code: Nx,
nutrient solution [N1, Yamazaki nutrient solution recipe; N2, BIO N (Swiss
Hydroponics Lab); N3, BIO NK (Swiss Hydroponics Lab)]; Ex, electrical conductivity
(EC) of nutrient solution [x, EC (mS cm-1)]; px, pH; Lx, LED tube (cool white, color
temperature 6500K) [x, light intensity (μmol m-2 s-1)]; dx, cropping density [x,
cropping density (plants m-2)]; Hx, duration of light period [x, light period (h day-1)];
AdT/nT, average day/night temperature (°C); W, water temperature (°C); DOx,
dissolved oxygen (ppm).
Pre-treatment stage Seedling stage Mature stage
Treatment
DBS 21 DAS 0-14 DAS 15-35/42
CK - N1_E1.2_p6.0_L200_d918_H24_A25 N1_E1.2_p6.0_L250_H16_d27_A25/18
BIO N N2_E0.4_W23_DO8 N2_E0.4_p6.0_L200_d918_H24_A25 N2_E0.4_p6.0_L250_H16_d27_A25/18
BIO NK N3_E0.4_W23_DO8 N3_E0.4_p6.0_L200_d918_H24_A25 N3_E0.4_p6.0_L250_H16_d27_A25/18

Yamazaki formula inorganic fertilizer was used as a control, and two organic fertilizers,
BIO N and BIO NK (trade names of Swiss Hydroponics Lab), were tested in this study. Lettuce
was grown hydroponically from sowing to harvest for 42 days (‘Frill-ice’) or 35 days (‘Boston’
and ‘Frilly’); fresh weight and minerals within the plants were then analyzed.

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Methods of measurement

1. Plant fresh weight.

Roots and sponge were removed, and plants were then weighed using an electronic
scale (HR-250AZ, A & D Company, Japan).

2. Nitrate and ammonium contents in nutrient solution.

A reflectometer (Rqflex 10, Merck) with nitrate (116971, Merck) and ammonium test
papers (116892, Merck) was used.

3. Mineral contents within plants.

One gram fresh leaf was taken and frozen at -20°C for 48 h, and sap was then extracted
using a centrifuge (z216MK, HERMLE), and then diluted to the appropriate concentration
(He et al., 1998). Samples were analyzed for anion and cation contents using an ion analyzer
(IA-300, DKK-TOA Corporation, Japan; cationic column PCI-205l; anion column PCI-322).

Statistical analysis
The results were analyzed using Duncan’s multivariate analysis using statistical
software SAS 9.1. Differences were considered significant when p<0.05.

RESULTS AND DISCUSSION

Saijai et al. (2016) used composted bark waste as the organic fertilizer in the water,
pretreated for 21 days and found that two types of beneficial bacteria, ammonia-oxidizing
bacteria (Nitrosomonas) and nitrite-oxidizing bacteria (Nitrobacter), were involved in the
conversion from ammonium to nitrite, and finally to nitrate. Shinohara et al. (2011) found
that, in some aquaponics systems, lettuce seedlings failed to grow, perhaps because there
was too much ammonium in the water. This reflects the lack of a pre-treatment stage. The
same conclusion was revealed in an early study by Atkin and Nichols (2004); they concluded
that an organic hydroponics system with only ammonium-nitrogen without nitrate-nitrogen
will significantly suppress lettuce growth.
As mentioned above, using an organic fertilizer in the nutrient solution requires pre-
treatment to allow ammonium- and nitrite-oxidizing bacteria to grow. As shown in Figure 1,
the ammonium concentration increased at the beginning and started to decrease at day 14,
while the nitrate concentration started to increase. The nitrate concentration of BIO NK on
day 21 was much higher than the nitrate concentration of BIO N on day 25.

Figure 1. Changes in nitrate and ammonium concentrations in organic fertilizers BIO N (A)
and BIO NK (B) during the pre-treatment stage.

Figure 2 shows three types of lettuce harvested under three types of nutrient solution.
Organic fertilizers seemed to suppress the growth of ‘Frill-ice’ lettuce. The leaf color seemed
to be lighter for ‘Boston’ and ‘Frilly’ lettuce grown using organic fertilizer. ‘Frilly’ lettuce

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grew better with BIO NK, with highest fresh weight, and ‘Frill-ice’ grew best with Yamazaki
formula. Lettuce grown using inorganic fertilizer led to higher nitrate content in the leaf, and
low nitrate content can be expected when grown using organic fertilizers, as shown in Table
2. Similar results were also found by Shinohara et al. (2011). This might be because of
interactions between the plant and microorganisms. In most cases, the plants grew slowly
with organic fertilizer compared with plants grow with inorganic fertilizers; however, the
human health benefit of greens with lower cellular nitrate-nitrogen may outweigh these
minor differences in yield. In this study, with the addition of a pre-treatment stage, as shown
in Table 2 and Figure 2, the fresh weight can be higher, and the merit of the low nitrate
concentration can be retained. Hanafy Ahmed et al. (2002) found that not only can
biofertilizers reduce the nitrate content but they can also increase total sugars, total free
amino acids and total soluble phenols in lettuce. It is suggested that plants may use the
accumulated nitrate as an osmoticum, enabling the plant to use more carbohydrates for
structural growth, thus increasing the dry matter.

Figure 2. Effects of different nutrient solutions on the growth of three cultivars of lettuce
(‘Frill ice’, 42 days after sowing; ‘Boston’ and ‘Frilly’, 35 days after sowing). Bar, 10
cm.

Table 2. Effects of different nutrient solutions on fresh weight and nitrate content of three
cultivars of lettuce. Means followed by different letters in each column are
significantly different at 5% level by Duncan’s multiple range test (n=10).
Fresh weight (g plant-1) Nitrate content (ppm)
Treatment
Frill ice Boston Frilly Frill ice Boston Frilly
CK 82.3a 88.8a 75.0c 6113.5a 3251.4a 5449.0a
BIO N 23.1b 64.1b 108.3b 888.4c 1333.6b 1159.0c
BIO NK 25.0b 92.1a 176.1a 1527.4b 3908.7a 2016.4b

As shown in Table 3, different nutrient solutions had no impact on ammonium

contents in leaves of lettuce. Organic fertilizers led to higher concentrations of Na, Ca and Mg
in leaves of lettuce. BIO NK led to higher potassium in the leaf compared with BIO N. Lettuce
grown in BIO NK had similar concentrations of potassium ions in the leaf compared to
lettuce grown using Yamazaki formula. As shown in Table 2, the fresh weight of lettuce
grown using BIO NK was much higher than that grown using BIO N. The reasons might be

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that BIO NK provides not only more nitrate, but also higher potassium and lower sodium
concentrations. Organic fertilizers can be further improved by reducing the concentration of
sodium.

Table 3. Effects of three nutrient solutions on various ions in leaf of three cultivars of lettuce.
Means followed by different letters in each column are significantly different at 5%
level by Duncan’s multiple range test (n=5).
Treatment Cultivar NH4 (ppm) Na (mg 100 g-1) K (mg 100 g-1) Ca (ppm) Mg (ppm)
CK Frill ice 7.4a 2.9d 384.5b 253.6c 98.8c
Boston 7.6a 3.0d 412.5ab 234.8c 73.5c
Frilly 15.0a 3.4d 418.0ab 328.0bc 87.2c
BIO N Frill ice 9.1a 18.6b 170.8d 840.0a 291.3a
Boston 10.8a 26.5a 262.0c 946.7a 296.3a
Frilly 16.8a 19.3b 138.8d 567.5b 216.7ab
BIO NK Frill ice 10.0a 8.8c 298.8d 192.7c 71.7c
Boston 10.5a 19.7b 484.0a 550.3b 185.6b
Frilly 10.3a 16.6b 403.5ab 546.7b 147.7bc

CONCLUSIONS
Using organic liquid fertilizer in PFAL is quite promising for semi-head and leafy green
lettuce production. Enhanced growth of root hairs and reduced nitrate concentration were
shown by previous researchers and in this study. This study provides further proof that, with
proper pre-treatment, it is possible to maintain the fresh weight of the produce. The
environment in a PFAL is relatively clean. The plant-microorganism relationship can be
properly managed to ensure efficient use of organic fertilizers. Most organic fertilizers can
be locally produced and extracted from agricultural wastes, thus making organic
hydroponics more environment friendly compared with traditional hydroponics using
chemical fertilizers. In the two types of organic fertilizer tested, BIO NK is preferred
compared with BIO N because of the rapid conversion of ammonium to nitrate, the rapid
growth of lettuce as well as the proper concentration of minerals in lettuce. Organic
fertilizers can be further improved by reducing the concentration of sodium. The need for a
pre-treatment stage seems to be the constraint in using organic liquid fertilizers if
production time is of great concern. More study will be conducted on the addition of
beneficial bacteria in organic hydroponics to reduce the duration of the pre-treatment.

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