Vol. 63, 2017 (4): 180–186 Res. Agr. Eng.

doi: 10.17221/47/2016-RAE

Extremely low frequency electromagnetic field generator

suitable for plant in vitro studies
Riry Prihatini1*, Mohamad Puad Abdullah2, Tuan Abdul Rashid
bin Tuan Abdullah3, Ismail Said3, Halil Hussin3, Norihan Mohamad
Saleh2,4
1
Indonesian Tropical Fruit Research Institute, Solok, Indonesia
2
Department of Cell and Molecular Biology, Faculty Biotechnology and Biomolecular
Sciences, University Putra Malaysia, Selangor DE, Malaysia
3
Department of Electrical Engineering, Faculty Engineering, University Tenaga Nasional,
Selangor DE, Malaysia
4
Agro-Biotechnology Institute Malaysia, National Institute of Biotechnology Malaysia,
Selangor DE, Malaysia
*Corresponding author: [email protected]

Abstract
Prihatini R., Abdullah M.P., Tuan Abdulllah T.A.R., Said I., Hussin H., Mohhamad Saleh N. (2017): Extremely low
frequency electromagnetic field generator suitable for plant in vitro studies. Res. Agr. Eng., 63: 180–186.

The extremely low frequency electromagnetic field (ELF-EMF) occurs naturally from the earth and artificially as a
human invention. The objectives of this study were to develop a suitable ELF-EMF generator for in vitro plants cul-
ture studies and to determine the effect of ELF-EMF exposure on in vitro tobacco (Nicotiana tabacum) growth and
chlorophyll content. An ELF-EMF generator, the coGEM 1,000 was constructed using four coils of copper wires that
were connected to a transformer, multimeter and rheostat. The coGEM 1,000 suitable for tissue culture plants is able
to produce stable and uniform 6 and 12 mT 50Hz ELF-EMF in the four coils of the ELF-EMF generator. The tobacco
in vitro plantlets were exposed to 6 and 12 mT of 50 Hz ELF-EMF for a period of 0.5, 1, 2 and 4 hours. The exposure
to 12 mT ELF-EMF for an hour increased plant growth (shoot height); whereas the exposure to 6 mT Elf-EMF for an
hour increased chlorophyll a, chlorophyll b and the total chlorophyll content.

Keywords: chlorophyll content; coGem 1,000; ELF-EMF; growth; tobacco plant

Electromagnetic field (EMF), a physical field con- particularly on seed germination have been re-
structed by a combination of an electric field and ported in several plants, viz. Arabidopsis thaliana,
magnetic field, influences the performance of a Prunus maritime, Phaseolus vulgaris, Cucumis
charged object in the surrounding area (Chrysiko- sativus (Pietruszewski et al. 2007; Pazur, Ras-
poulos 2009; Mrozynski, Stallein 2013). The sadina 2009; Yan et al. 2009). Investigations on
EMF produced from 30–300 Hz electric current is the effect of exposure of these plants to ELF-EMF
generally referred as extremely low frequency EMF were performed in the presence of a wide range of
(ELF-EMF). The effects of the ELF-EMF on plants, electromagnetic strength, ranging from 0.7 μT to

Supported by the Indonesian Tropical Fruit Research Institute, Indonesian Agency of Agriculture Research and Development,
and Ministry of Agriculture.

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60 MT at a short duration of 10 min to an extended and parallel. Overall there were five designs that
duration of 56 days (Calestino et al. 1998; Mon- were tested. For each design, the functional test was
selise et al. 2003; Odhiambo et al. 2009; Yan et carried out to determine the capacity and uniform-
al. 2009). Furthermore, the frequency of ELF-EMF ity of the ELF-EMF strength that can be generated
applied in these studies were also varied, viz. 50 Hz, from each coil on the circuit. The best design was
60 Hz, 65 Hz, 75 Hz and 100 Hz (Reed et al. 1993; the one that had capacity to produce a uniform ELF-
Monselise et al. 2003; Pazur, Rassadina 2009). EMF strength up to 12 mT. Following that, a stability
Compared to the research of the ELF-EMF effect test was conducted on the best circuit to determine
on animals and humans, to date, only limited studies the stability of the ELF-EMF produced by each coil
on the effects of ELF-EMF on in vitro plants have when the generator operated for 5 hours. The Gauss
been documented (Goldsworthy 2006), where meter logger data were employed to record the ELF-
the stability and uniformity of the ELF-EMF ap- EMF that was produced by each coil. The observa-
plied to the in vitro plant subjects were not clearly tion data of the ELF-EMF strength versus time were
described. The ELF-EMF generators commercially translated into graphs using the EMCALC computer
available at present are generally based on the Helm- software (Enertech Inc., USA).
holtz coil concept. This rather bulky device is often Preparation of plant material. The tobacco
able to negate external magnetic field surrounding it mother plants as the explants source for this ex-
while being able to create the electromagnetic field periment were germinated from seeds of Nicotiana
within the coil. Using a similar principle, this study tabacum on a MS medium (Murashige, Skoog
developed an ELF-EMF generator that can be used 1962). The stem explants (3 cm) were obtained
as a small-scale generator for in vitro plants studies. from 4 weeks old mother plants. Four-day-old cul-
This generator, named coGEM 1,000, was built using tures were placed in a vessel (a glass jar with di-
several copper wire coils, which is upgradable, and ameter of 7 cm and height of 12 cm). The in vitro
was connected to other electrical components to en- tobacco plants were then subjected to two different
sure accuracy, stability, reproducibility of the ELF- strengths of ELF-EMF (6 and 12 mT) at four differ-
EMF generated and safety of the system developed. ent durations of exposure (0.5, 1, 2 and 4 hours).
As a model plant for biological research and plant The plants of similar age which were not exposed
biotechnological applications (Ganapathi et al. 2004; to ELF-EMF, but were grown under similar condi-
Budzianowska 2009), Nicotiana tabaccum (tobacco tion, were chosen as controls. After the exposure,
plant) was used in this study to assess in vitro plant re- the in vitro cultures were incubated in the growth
sponse to the ELF-EMF exposure at specific strength chamber with a 16 h photoperiod at 25°C.
known to have a positive effect on plants (Dardeniz Assessment of growth parameters and chloro-
et al. 2006, Huang, Wang 2007; Yan et al. 2009; phyll content. The growth parameters were meas-
Shabrangi et al. 2013). The present study reports on ured to find the effect of the ELF-EMF on plant
the growth, morphological development and chloro- growth and developments. The chosen parameters
phyll content of the tobacco plant in response to the were namely number of shoots, shoot height and
ELF-EMF generated by the coGEM 1,000. number of leaves. The number of shoot and leaves
was counted manually, while shoot height was de-
termined by measuring the stem length from the
MATERIAL AND METHODS points where roots occur until the end of the stem.
The chlorophyll content of the plants was anal-
Construction of the ELF-EMF generator. Setup ysed using the modified method proposed by NI et
of the ELF-EMF generator was started by connect- al. (2009), which can be described as follows: ap-
ing 1,000 turns of copper wire (AW16) that was proximately 100 mg of fresh leaves were ground
coiled around the PVC core (coil 1) to the power into a fine powder in the presence of liquid nitrogen.
supply, a transformer, and a multimeter (Tektonix The powdered leaves were transferred into a 1.5 ml
DMM 4040; Tektonix Inc., USA) (referred as circuit microcentrifuge tube covered with aluminum foil
design 1). The design was gradually expanded by and added with 1 ml of 80% acetone. The microcen-
adding another coil into the design, one at a time. trifuge tube was then centrifuged at 4°C for 15 min
The circuit also modified the series connection, par- (3,000 g), and the supernatant was transferred to a
allel connection and combination between series new centrifuge tube and kept in the dark. The ab-

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sorbance (A) of chlorophyll content in the mixture showed that the electromagnetic field produced had a
was measured using a spectrophotometer at 663, linear correlation with the current input. Several coils
645 and 470 nm wavelength; and 80% acetone was arrangements (series, parallel, and combination of se-
used as a blank control. The chlorophyll concentra- ries and parallel) were tested and the data obtained
tion of each sample was then calculated as follows: were plotted. The parallel and series circuit (fourth
Chlorophyll a content (mg/g) = [(12.7 × A663) – circuit) generated 60 Gauss of magnetic field from 2.6
(2.69 × A645)] × V/1,000 × W A current and 120 Gauss from 5.3 A current in every
Chlorophyll b content (mg/g) = [(22.9 × A645) – coil. Furthermore, it was also revealed that all coils
(4.86 × A663)] × V/1,000 × W had similar current and electromagnetic field. Follow-
Total chlorophyll content (mg/g) = [(8.02 × A663) + ing the construction of the CoGEM 1,000 (Fig. 1), the
(20.20 × A645)] × V/1,000 × W stability of electromagnetic field was also analysed.
Carotene/xanthophyll content (mg/g) = [(1,000 × The experiment was conducted in order to determine
A470) – (3.27 × Ca) – (1.04 × Cb)]/229 if there were any variations in electromagnetic field
Ratio of chlorophyll a and b = Ca/Cb produced in each coil in the generator.
A 663 – absorbance on 663 nm; A645 – absorbance on
645 nm; A663 – absorbance on 470 nm; V – volume of
the extract (ml); W – weight of fresh leaves (g); C a – Effect of the ELF-EMF on the tissue-cultured
concentration of chlorophyll a content (mg/g); Cb – con- tobacco plants
centration of chlorophyll b content (mg/g)
Experimental design and statistical analysis. All The statistical test of the effects of ELF-EMF on
in vitro experiments were arranged in the Randomly developmental parameters of tobacco plants re-
Complete Block Design (RCBD) which included 8 vealed that ELF-EMF exposures to the tissue-cul-
treatments and one control. There were 20 samples tured tobacco had significantly affected (P < 0.05;
for each treatment; the whole experiments were F Wilk’s Lambda = 10.394) the shoot height of the
replicated three times. The data were analysed us- plants, whereas the treatment did not cause any
ing a multivariate analysis of variance (MANOVA) changes on plant’s number of shoots and leaves.
that was based on the Wilks’s Lambda to identify the The treatment of the 12 mT ELF-EMF for 0.5 hours
significance between groups of treatment (P < 0.05). (3.9 ± 0.8 cm) increased shoot height by 44.4%; yet a
Statistical analyses were done using the computer higher and longer duration of ELF-EMF exposures
software of SPSS ver. 18.0 (SPSS Inc. 2009). (more than 12 mT 0.5 hour treatment) significantly
reduced shoot height.

RESULTS
Effect of ELF-EMF on the chlorophyll content
Development of the ELF-EMF generator of tissue-cultured tobacco plants

The electromagnetic field required for this study was The MANOVA result in an effort of finding
120 Gauss (equivalent to 12 mT). The data calibrated out the effects of ELF-EMF exposure to the to-

Fig. 1. The CoGEM 1,000, an ELF-EMF generator consists of 4 coils connected in series and parallel with AC supply,
ampere meter and rheostats

182
 1.5 1.5 25

Shoota
Shoot
1.0 20

(mg/
1.0

Chlorophyll
0.5 0.5 15
0.0 0.0 10

12 mT 4 h
Control

6 mT 0.5

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h

12 mT 2 h
12 mT 2 h

12 mT 4 h
Control

6 mT 0.5

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h
5
0
Res. Agr. Eng. Vol. 63, 2017 (4): 180–186

12 mT 2 h

12 mT 4 h
6 mT 4 h

12 mT 0.5 h

12 mT 1 h
6 mT 1 h

6 mT 2 h
Control

6 mT 0.5 h
ELF-EMF Exposure ELF-EMF Exposure
doi: 10.17221/47/2016-RAE
ELF-EMF Exposure

4.5 d

a content
bcd cd 35 b b b
Chlorophyll a content

35
4.0 c b
height (cm)

a ab ab a ab
3.5
30 b bc 30

b content
45 c c bc bc c
3.0
25 25
40 b b

(mg/g)
b
Shoots(mg/g)

2.5 20
35 a
20 a a a

Chlorophyll
Chlorophyll
2.0 30

(mg/g)
15 15
1.5 25

Chlorophyll
10
1.0 10
20
0.55 5
15
0.00 10
0
5

12 mT 2 h

12 mT 4 h
h 4h
12 mT 0.5 h

12 mT 1 h
1h 1h

h 2h
Control

0.5 h0.5 h
mT22hh

mT44hh
Control

0.50.5

mT11hh

mT22hh

mT44hh

0.5hh

mT11hh
Control

mT0.5

mT
6 mT

mT
6 mT

12 mT 1 h

12 mT 2 h

12 mT 4 h
Control

12 mT 0.5 h
66mT

66mT

66mT

12mT

12mT

12mT

6 mT

62

64
6 mT

12mT

6 mT

6 mT

6 mT
12

12

12

6 mT
12

ELF-EMF
ELF-EMFExposure
Exposure ELF-EMF Exposure
ELF-EMF Exposure

b content
45
Chlorophyll b content

45 c c bc bc c 40 c c bc bc c
bcd cd
a content

4035 b b 40 d bd b
b b ab

chl orophyll
35 aab c

chlorophyll
3530 a 35 a

(mg/g)
30
30

content(mg/g)
3025
(mg/g)

25
25
(mg/g)

25

Chlorophyll
Chlorophyll

2020 20
20
Chlorophyll

1515 15
15
1010 10
10
Total 55
5
0 5 00

mT44hh
Control

0.5hh

mT11hh

mT22hh

mT44hh

0.5hh

mT11hh

mT22hh
0
12 mT 2 h

12 mT 4 h

Control
Control

6 mT 0.5 h

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h

mT0.5

mT0.5
12 mT 2 h

12 mT 4 h
6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h
Control

6 mT 0.5 h

6 mT 1 h

66mT

66mT

66mT

12mT

12mT

12mT
66mT

12mT

12
12

12
12
ELF-EMF Exposure ELF-EMF
ELF-EMFExposure
Exposure
ELF-EMF Exposure

40 40 cd cd d d
cd cd d d
b ab
Total chl orophyll

ab 35 ab c a
Total chl orophyll

35 ab c a
content (mg/g)

b
(mg/g)
b content

1.4
30 45 30
c c c abcbc bcbc c 25
2540 b
a:b

1.2 b ab b bb
a:b

20
1.0 35 a ab a a Fig. 2. Effect
20 of different strengths and duration exposure
Ratio chl

1530 15
content
(mg/g)

0.8 to ELF-EMF (extremely low frequency electromagnetic

1025 10
Chlorophyll

0.6
520 5
field) on the pigment content of the tissue-cultured to-
0.4015 0
0.210 bacco plantlets 4 weeks after culture; (a) shoot height, (b)

12 mT 4 h
12 mT 1 h

12 mT 2 h
Control

6 mT 0.5 h

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h
h 4h
Control

mTh0.5 h

h 1h

h 2h

h 4h

mTh0.5 h

h 1h

h 2h

5
0.0 0
chlorophyll a content, (c) chlorophyll b content, (d) total
6 mT6 1mT

6 mT6 2mT

6 mT6 4mT

12 4mT
12 1mT

12 2mT
Control

12 mT 4 h
Control

6 mT 0.5 h

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h

12 mT 2 h
6 mT6 0.5

12 0.5

chlorophyll content, and (e) ratio of chlorophyll a/b

12 mT

12 mT

12 mT
12 mT

ELF-EMF Exposure
mean ± standard error followed
ELF-EMFby the same letter are not
Exposure
significantly different from each other according to the
ELF-EMF
ELF-EMFExposure
Exposure
Duncan’s multiple range test at P = 0.05

40 cd cd d d
b ab
Total chl orophyll

35 ab c a
content (mg/g)

bacco plant30 biochemical properties indicated that DISCUSSION

25
the ELF-EMF20 exposures had a significant effect
(P < 0.05, 15
F Wilk’s Lambda = 6.203) on the con- Influence of the circuit design on capacity
10
tent of chlorophyll
5 a, chlorophyll b, total chloro- and stability of the generated ELF-EMF
0
phyll and ratio of chlorophyll a/b. The exposure to
12 mT 2 h

12 mT 4 h
Control

6 mT 0.5 h

6 mT 1 h

6 mT 2 h

6 mT 4 h

12 mT 0.5 h

12 mT 1 h

the 6 mT ELF-EMF for an hour on tobacco plant- In order to carry out this study, an ELF-EMF
lets produced the highest content of chlorophyll a generator that can produce the specific ELF-EMF
(32.3 ± 0.14 mg/g), chlorophyll b (37.6 ± 0.98 mg/g) strength needed to be developed. Several factors
ELF-EMF Exposure
and total chlorophyll (20.4 ± 0.27 mg/g) or in- were necessary to be considered in the develop-
creased by 17.9%, 41.7% and 17.8%, respectively. ment of the ELF-EMF generator, namely the num-
However, the ratio chlorophyll a/b was decreased ber of turns on the coils, number and size of the
since the increment of chlorophyll b content was coils, the amount of electric current needed and
higher that the increment of chlorophyll a content safety of the equipment to the operator and the en-
(Fig. 2). vironment.

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An ELF-EMF generator which consisted of four magnetic field inside each coil when the current of
coils (which refer to 4 chambers) was built in order 0.5 to 5.3 Ampere (at 0.5 A steps) was applied.
to investigate the effects of ELF-EMF on tissue-cul-
tured plants. The currently available ELF-EMF gen-
erators were usually not appropriate for this study The coGEM 1000
because their size was too big, thus it required more
expensive materials to be constructed, whereas the The functional and stability tests for the parallel and
ELF-EMF generated by such generator would be series circuit (fifth circuit) showed that the ELF-EMF
less uniform and constant. The common ELF-EMF generated from the coGEM 1,000 was up to 12 mT
generator also required high electrical voltage and and it can be operated for 5 hours steadily, continu-
current input, making it unsafe to be operated in ously and safely. This means that the circuit is an excel-
the plant tissue culture laboratory. lent ELF-EMF generator and it is capable of produc-
Initially, only a single chamber or coil was con- ing high and uniform electromagnetic field in every
structed. After that, the second and the third coils coil. This generator was then named coGEM 1,000
were added in series. When there was only one coil, and was used for the following experiment to find
the result showed that the coil could produce 12 mT out the effects of ELF-EMF on the rate growth and
ELF-EMF. However, in the presence of the two coils chlorophyll content of tobacco plants.
in series, the ELF-EMF produced by both coils was This coGEM 1,000 is a suitable ELF-EMF gen-
not uniform. The second coil had different resist- erator that could be applied in the tissue culture
ance compared to the first coil, thus causing the study since it has many advantages. First of all, the
variation in the electromagnetic field produced by coGem 1,000 is a user friendly as it is easy to oper-
the second coil. ate; the ELF-EMF strength can be varied by setting
When three coils were connected in series, the up the current input. It was also easy to calibrate it
three coils generated different and smaller electro- by measuring the ELF-EMF produced by each coil
magnetic fields compare to single and two coils ar- with the common Gauss meter and the intact soft-
rangements. Connecting third coil resulted in the ware. The coGEM 1,000 built is a basic equipment
formation of a weak electromagnetic field. Even with which can be upgradable by adding another coil
the highest current possible to be generated from into design and by modifying the circuit to con-
the circuit (1.8 A), coil 1 only emitted 94.08 Gauss, struct larger generators for mass application.
coil 2 emitted 89.8 Gauss, while coil 3 emitted
76.54 Gauss. It was clear that the circuit had some
flaws, and therefore it could not generate the uni- Effect of the ELF-EMF on the tissue-cultured
form magnetic field which had different resistance tobacco plants
among the three coils.
Thus, a parallel connection of the coils was pro- Most studies on the impacts of ELF-EMF on various
posed to overcome the electromagnetic field incon- crop plants have been conducted using native plants
sistency produced by the generator. Coils 1 and 2 that are grown under natural conditions. However, it
were connected in parallel with the AC power sup- is difficult to measure the true impacts of ELF-EMF
ply and multimeter. The magnetic field produced on these plants since other abiotic factors contrib-
inside both coils from 0.4 to 6.0 A (in 0.4 A steps). ute to the measurement. In order to eliminate these
The data were recorded, and the graph was plotted. factors so that the true impacts of ELF-EMF on the
The main consideration on the construction of the plants can be precisely measured, in vitro culture also
ELF-EMF generator was to ensure that the flow of known as “tissue culture system” was selected for this
the current in each coil was uniform so that it could research. In this study, selected plants which had been
generate the same electromagnetic field in all four propagated through the tissue culture technique were
coils. To achieve that, the following circuit was de- used to study the impacts of ELF-EMF on plants.
signed and included the additional apparatus known High shoots were also induced in cucumber and
as rheostat (variable resistor). The four coils were chilli seedlings when exposed to lower ELF-EMF
connected in parallel connection of two series coils strength (0.1 mT and 62 μT, respectively). Expo-
with the AC supply, a multimeter and two rheostats. sures of etiolated Cucumis sativus seedlings to
The circuit was calibrated by measuring the electro- 50 Hz 1 Gauss ELF-EMF produced higher shoots

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(500%) compared to the unexposed seedlings (Pia- Similar to the ELF-EMF, approximately a two-
centini et al. 2001). fold increase in the concentration of chlorophyll a
and total chlorophyll content was observed when
PGR cytokinins (0.4 µM 4-PU-30 [N-phenyl-N’-(-
Effect of ELF-EMF on the chlorophyll content chloro-4-pyridyl) urea], urea and TDZ) were added
of tissue-cultured tobacco plants in the MS growth medium of 40-day-old of Dian-
thus caryophyllus plantlets (Genkov et al. 1997).
Due to its function to absorb solar radiation, the Thus, the application of ELF-EMF on tissue culture
chlorophyll content could be directly estimated using technique can potentially replace the function of
a photosynthetic potential and primary production. plant hormone as a plant growth inducer.
Furthermore, chlorophyll content can also be used to In an in vitro system, plants are often characterized
resolve indirect determination of the nutrient level, by a low photosynthetic rate since the availability of
since most of the nitrogen is integrated in chlorophyll sucrose on the media allows plants to use the carbon
(Gitelson et al. 2003). Chlorophyll content is also source from the medium instead of producing car-
associated with the physiological state, such as stress bohydrate through photosynthesis (Kadlecek et al.
and senescence (Damaraju et al. 2011). 2003). However, the increase of chlorophyll content
On the one hand, the chlorophyll content affects the of the in vitro plantlets might be an advantage since
carbon dioxide exchange rate of photosynthesis, and it can increase the survival rate of the plants in the
thus it is also affected by other physiological process- acclimatization stage (Afreen 2005).
es, such as mitochondrial respiration and photorespi-
ration (Tanaka, Tanaka 2011). On the other hand,
the chlorophyll content can be stimulated by endoge- Conclusion
nous cytokines, irradiance, temperature and drought
stress (Olah, Masarovikova 1998; Afreen 2005). The device used to generate the ELF-EMF was
Chlorophyll contents, including chlorophyll a, successfully constructed. The complete set of the
chlorophyll b, total chlorophyll a and b and carotene generator is called coGEM 1000 and consists of four
were used in several studies as biochemical param- chambers, each made up of copper wire coils con-
eters. For instance, tobacco leaves from a plantlet nected to the transformer, multimeter and rheostat.
with high irradiance treatment showed a decrease in The four chambers or coils generator can achieve a
total chlorophyll content and chlorophyll a/b ratio, maximum of 120 Gauss or 12 mT ELF-EMF at 5.3 A
but the contents of carotene and xanthophylls pig- current when the coils were connected in a parallel
ment were increased (Kadlecek et al. 2003). In in connection of two series coils. The ELF-EMF pro-
vitro plant culture, chlorophyll content might not be duced by the equipment was tested for its stability
affected by media concentration, but increasing the and uniformity. The result showed that it can pro-
amount of air exchange in the culture vessel would duce stable 6 and 12 mT ELF-EMF for 5 hours. The
increase the chlorophyll contents (Afreen 2005). result obtained showed that the ELF-EMF exposures
The induced chlorophyll concentration in the had a significantly increasing shoots height of tobac-
tobacco plants exposed to the ELF-EMF might be co cultures. It was also observed that the exposure
triggered by the increased activity of the chloro- increased chlorophyll a, chlorophyll b, and the total
phyll synthesis-related enzymes. The chlorophyll chlorophyll content of the plant. Thus, these results
biosynthesis started with glutamate that was oxi- implied that the ELF-EMF exposure had a potential-
dized into several intermediate compounds before ly positive effect to increase growth and chlorophyll
converted into chlorophyll a or b. The pathway content of plant in vitro culture.
involves many specific enzymes, coenzymes and
cofactors, such as photochlorophyllide oxireduc- References
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Received for publication April 19, 2016
Accepted after corrections September 7, 2016

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