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UDC 502/504



Mateusz Jakubiak1, KatarzynaGdowska2

PhD. Eng., assistant professor, AGH University of Science and
Technology, Faculty of Mining Surveying and Environmental Engineering, 30
Mickiewicza Av., 30-059 Krakow, Poland
MSc. Eng., assistant lecturer, AGH University of Science and
Technology, Faculty of Management, 30 Mickiewicza Av., 30-059 Krakow,

Interdisciplinary studies have pointed out many possible uses of laser

biotechnology in environmental engineering and agriculture. A number of previous
research studies on the application of low intensity laser stimulation of different
biological material proved, that effects of photostimulation may increase biomass
production, yield and germination energy. Laser stimulationmay also increase
plants resistance to environmental pollutions and unfavorable environmental
factors. Methodmay be used also for more efficient sewage treatment process, soil
reclamation and increase of bioremediation abilities. Application of laser
biotechnology might be use in environmental engineering technologies, according
to ideas of sustainable development.
Laser stimulation, willow biomass, calorific value , Salix vimnialis.

Introduction Application of laser stimulation.

Poland has signed a number of international conventions on environmental
protection. European Union environmental guidelines require Poland to increase
the share of renewable energies in the overall fuel-energy balance of the country.
Authorities have issued a number of legislative documents and regulations to
promote and support the production of green energy. In Poland, among many
sources of renewable energy, the most popular is biomass. However, Polish forests
do not have enough firewood to meet the needs of individual customers and power
plants. The development of agriculture oriented on energy crops has increased the
possibilities for using wastelands and brownfields. Energy plantations of fast
growing biomass plants are becoming more widespread way of agricultural land
use. Tree species (e.g. willows, poplars), perennial dicotyledons (e.g. Jerusalem
artichokes, Virginia mallow) and perennial grasses (e.g. miscanthus) are the most
commonly grown energy crops. Willow (Salix viminalis) is the most popular
energy plant. Farmers cultivating willows on an industrial scale search for methods
to increase the biomass yield. Several studies were conducted on laser stimulation
or mycorrhization of willow cuttings.
Research studies on the application of laser photostimulation of different
biological material for improved protection of the environment were initiated in
mid 1970s. The basic idea was stimulation of physiological processes of adaptation
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to environmental changes without changing organisms genotypes. Scientific
research lead by Prof. Dobrowolski team proved, that the method is successful in
increase of energetic status as well as homeostatic abilities of organisms for better
adaptation to suboptimal environmental conditions. Stimulation of plants may also
increase their resistance to environmental pollutions and unfavorable
environmental factors. Interdisciplinary research studies on the application of low
intensity laser stimulation that effects of photostimulation by coherent light are
related to the quantum of energy absorption of laser radiation by certain
biologically active compounds or cellular organelles. The effect of radiation
absorption by cells could increase certain enzymes activity; mainly those involved
in the cells energetic processes. Laser light stimulation may result in a change in
cells metabolism. Research studies proved that the effectsofa monochromatic laser
light on the biological material have non-specific character. During the last
decades it has been found, that the effects of a monochromatic laser light on the
biological material have non-specific character. The light is able to cause effects of
biostimulation at different organisms like bacteria, algae, protozoans, fungi as well
as at plants and animals as well as at in vitro animals and plant cells cultures and in
vivo vascular plants cells cultures [4, 5, 6, 12, 14].
Results of a number of prior researches on monochromatic, coherent laser
light biostimulation prove that the method is successful in increase the usefulness
of some plants for reclamation. Stimulation of plants may increase biomass
production and plants resistance to environmental pollutions and unfavorable
environmental factors. Moreover, it could change the amount of trace elements
accumulated in plant tissues [4, 5, 24]. It was found that Salix viminalis irradiated
with argon laser have ability to boost concentration of some elements in biomass.
Average accumulation of Cu, Cr, Mn and Fe rose twice while deposition of Zn and
Pb was about 50% higher when compared to unirradiated, control material [24,
27].Industrial plants like the fast-growing varieties of energetic willow such as
Salix viminalis are appropriate for use in bioremediation of chemically
contaminated lands. Harvested willow biomass may be used as a material for the
production of energy in the combustion process.Experiments conducted on the
willows (Salix sp.) which cuttings were exposed to light form argon laser, helium-
neon laser and laser diodes, have shown that stimulation may affect the
rhizogenesis acceleration and faster growth of shoots. Increased resistance to
stress-exposed plants, better rooting, a larger biomass increase and the extension of
the growing season were noted as consequences of laser photostimulation of
willow seedlings [27].
Research conducted on the agricultural use of laser light show that pre-
sowing laser stimulation could be a factor increasing seed quality and accelerating
the early phase of growth. However, the pre-sowing biostimulation of crop plants
seeds has a greater impact on crops of vegetables such as cucumbers, tomatoes,
lettuce, and eggplant than on grain crops. A more synchronous germination,
accelerated ripening and a higher yield is a positive effects of vegetables seeds
irradiation. Moreover, a higher biological value of fruits and an increased content
of nutrients is an important result of the photostimulation of agricultural
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production. Low intensity laser stimulation of seeds and seedlings causes the
absorption of light energy. Absorbed energy is converted into chemical energy,
which is used for growth and development of plants. The provision of energy
increases the energy potential of the seeds, which affects the physiological
processes in the germinating seeds. Possible effects include better and faster
germination, accelerated ripening and increased disease and frost resistance [8, 9,
13, 14]. Appropriately selected parameters of photostimulation can also increase
the percentage of germinating seeds. Photostimulation of garden cress (Lepidium
sativum) seeds may significantly contribute to immunization of germinating plants
to adverse conditions caused by high salinity [15].
Research on the use of stimulation monochromatic, coherent laser light on
seeds and seedlings of crops to increase their yields are widely carried out in a
number of scientific centers in the world. The impact of laser irradiation was
studied for a number of vegetables and cereals e.g. peppers, soybean, carrots, sugar
beet, tomatoes, cucumbers, onion, peas, potatoes as well as anise, cumin, white
lupine, lucerne, white and red clover , maize, spring wheat, barley, oilseed rape.
The possibility of germination increase of soybean (810%), peppers (314%),
carrot (5%) and sugar beet (12%) by He-Ne laser stimulation of seed was shown in
studies conducted in Bulgarian research centers [1]. The Egyptian studies on
soyabean seed treated by power of the green laser source with wavelength 532 nm
(Nd-Yag laser) have shown that seeds earlier emergence during germination [10].
It was noted that in the roots of sugar beets grown from laser stimulated seeds the
sugar content was increased about 1517% [13]. Furthermore, the stimulation of
germination and embryoroot length of pepper were observed as effects of laser
light irradiation [22]. The increase in the yield depends on the stimulation
parameters used for irradiation of selected vegetables. The increase of crop yields
depends on the vegetables type: tomatoes 20 - 24%, peppers 13%, cucumbers 15%,
onion 15.5% and beans 27%. An important parameter used in the production of
vegetables is the percentage of faster ripening plants. It could be increased for
peppers (19%24%) and cucumbers (17%). As to, dry weight, it increased by 3.8%
in the case of beans, 5.3% for pepper and 6.5% for onions. The presented effects
resulted from better health of plants, generally higher degree of tolerance to
physiological stress, better growth and development of plants treated with laser
light [25]. Another research conducted in Egypt confirmed the effectiveness of He-
Ne laser stimulation of crop plants seeds such as anise (Pimpinella anisum) and
cumin (Cuminum cyminum). The increase in yield and increased juice content in
the plant cells resulted in increased quantities of essential oils from plants up to
165% of anise and 140% of cumin [11]. Further positive impact of the laser
stimulation on the vegetable crops was reported in potatoes yields. The potatoes
tubers were irradiated before planting. The average potatoes yield increased by
43.1%. The increase in yield was caused by increase of the tubers number from
single plants by 17.8% and an increase in the average weight of single tubers by
21.4% [26].
Conducted studies also confirm the positive effect of laser stimulation on
green plants and cereal. White lupine (Lupinus albus) plants grown from seeds
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stimulated by laser light had greater leaf area than plants grown from control
unirradiated seeds [17]. Many experiments which aim was to determine the effect
of coherent light stimulation on the growth and yield of corn seeds were described
in the literature. It was shown that irradiation of maize seeds (Zea mays) modifies
the initial growth and development of maize seedlings. The observed average
increase in length of shoots and roots of seedlings grown from irradiated seeds was
respectively - 42.9% and 64.6%. Furthermore, irradiated maize seeds weight
enlarged rapidly during swelling, when compared with unirradiated seeds [18]. It
was found that the use of pre-sowing irradiation of seeds with laser light could also
lead to a productivity increase of some crops cereal, e.g. maize (1015%), spring
wheat (2030%), barley (2025%), rapeseed (1015%). The cereal crops quality
sown from the irradiated seeds was also better than in the control group. For
example, the protein content in grain of spring wheat grown from seeds stimulated
by He-Ne laser increased about 1214%. Research on the pre-sowing stimulation
with laser light of winter wheat seed showed an increase of germination energy and
in percentage of germinating seeds [16]. A research on 12 days old seedlings of
spring wheat (Triticum aestivum) was also carried out. Results showed that a
proper light stimulation of seedlings, with use a CO2 laser, increased resistance to
wheat germ stress caused by water scarcity. Further observations have shown that
laser stimulation had long-lasting, positive impact on growth of seedlings during
drought [19].
It is worth noting that equipment to automate the process of pre-sowing laser
stimulation of seeds was developed in a number of research centers. One of the
examples of application of pre-sowing laser seeds stimulation by divergent beam
method is a Polish device developed and patented by Dygdaa and Koper [7].
SOLAR-2M, yet another device for seed photostimulation, was constructed by a
team of Bulgarian Academy of Sciences in 2005 [2, 3].
The purpose of research. The presented experiment was part of a broad
research on selected energy willow varieties (Salix sp.). The aim of this study was
to ascertain whether the selected laser stimulation parameters of willow cuttings
could improve their physical fitness as well as enhance and accelerate the
production of biomass. The purpose was to verify the influence of
photostimulation method on the chemical and technical parameters as well as
calorific value of willow wood.
Material and methods of research. The parameters of irradiation were
chosen on the basis on the results of preparatory hydroponic experiments, previous
field experiments and on studies on other plant material. Photostimulation
algorithms were based on the following parameters: wavelength, energy density,
duration and mode of exposure.
Some species of energetic willow could be suitable for cultivation on
contaminated areas like wastelands and postindustrial. These species are resistant
for diseases and adverse environmental conditions. Large and rapid growth of
biomass is an important advantage of these varieties. Especially, when willows
wood is use as an energy source. The biological material used in this experiment
were Salix viminalis Duotur cuttings. This polish variety, resistant to disease and
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tolerant to pests, was created at The Chair of Plant Breeding and Seed Production
of The University of Warmia and Mazury in Olsztyn. Additionally, it gives a very
high yield of dry wood which is why the biomass collected after a vegetation
season biomass can be used as a source of renewable energy.
Particular experimental groups of willow cuttings have been exposed to light
emitted by the following devices:laser diode, produced by Changchun New
Industries Optoelectronics Tech Co., emitting light of wavelength corresponding to
blue color =473 nm and with power of 20 mW and impulse medical laser, type
Laser D68-1, produced by Marp Electronic, emitting pulsatory light of
wavelength corresponding to red color =670 nm and with power of 20 mW.
There were 30 cuttings prepared per group. Seedlings in the experimental
groups were exposed to laser light with different parameters before planting.
Irradiation was intermittent. The distance from the light source to the biological
material was 20 cm and the light beam was perpendicular to the surface of the
material. The cuttings used in experiment were divided into three groups:
- control group of unirradiated cuttings, marked as C,
- experimental group, irradiated by laser diode, total exposure time of 90
seconds (3 sequences in 30 seconds), marked as DN,
- experimental group, irradiated by impulse medical laser, total exposure
time of 90 seconds (3 sequences in 30 seconds), marked as LM.
After the second growing season willow shoots were collected. Wood
samples of each group, prior to the calorimetric analysis, have been dried at room
temperature. The analyses of carbon, hydrogen and sulfur content of the wood
have also been made.
The analyses of the suitability of biomass for energy purposes is based on
the following standards: PN/G-04511 analysis of moisture content, PN/G-04533
- the ash content was assessed by the classical method of burning at a temperature
of 815C, PN-ISO 562: 2000 the volatile matter content was performed at 900C,
PN-ISO 351:1999 - total sulfur content and ash sulfur content, PN/G-04521 - the
contents of carbon and hydrogen in the biomass are estimated by the high fuel
combustion, PN-ISO 1928: 2002 - the heat of combustion and calorific value.
The results of research. The results of experiments can make a basis to
create optimal photostimulation parameters of willow cuttings for enhancement
and acceleration the production of biomass. Significant differences in the increase
of biomass in experimental groups were found. The average weight of harvested
shoots from the control group (C) was 121.4 g (SD 63.5 g), group DN 205.1 g (SD
101.9 g) and group LM 168.0 g (SD 94.3 g). In comparison with the control
group differences in weight of shoots in experimental groups DN and LM are
statistically significant.
Chemical, technical parameters and calorific value from a control group and
experimental groups were measured and compared (Table).

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Table. Technical, calorimetric and chemical analysis of harvested
biomass from control (C) and experimental groups: DN, LM
Group C DN LM
Technical analysis
Freemoisture (Wex) 49,2 48,4
Moisture (W) 8,4 7,9
Ash (A) 2,0 1,4
Volatilematter (V) 74,26 74,90
[%] 74,40
Calorimetric analysis
Calorific value Qi 16881 16643
-1 16839
[kJ*kg ]
Chemical analysis
Carbon (C)
[%] 47 46,3 48,1
Hydrogen (H)
[%] 5,39 5,38 5,42
Sulfur (S)
[%] 0,20 0,25 0,22

The results of experiment confirmed that stimulation by low power lasers
with parameters of irradiation properly chosen for the Salix viminalisDuotur can
significantly speed up growth of plants. Results clearly indicate that the best
growth may be achieved using stimulation by laser diode emitting wave length:
=473 nm, with irradiation total time of 90 seconds (3 sequences in 30 seconds). A
better effect of biomass growth then in control group was achieved in experimental
group irradiated by impulse medical laser.
The parameters important for biomass use for energy purposes are very
similar in all groups. Comparing with the data from other studies, calorific value is
on the standard level for one-year shoots of willows. However, the ash content in
all groups is very low. The one-year-old willow shoots wood has calorific value -
17 008 kJ*kg-1 and ash 6.89% [20]. The calorific value of dry biomass of others
one year old shoots energetic plants is lower. For example, Virginia fanpetals (Sida
hermaphrodita Rusby) stems has 15100kJ*kg-1[23]. The literature data published
by researchers working on the energetic willows show that using a suitable system
of plants cultivation it is possible to increase calorific value. In addition, willow
crops harvest on a three-year rotation may result in calorific value of almost 20000
kJ*kg-1 [21].
Observations allowed to conclude that properly selected parameters of
coherent light stimulation may effect infaster biomas growth without changing the
chemical or technical parameters and the calorific value of wood. The results
reaffirm the need to continue studies on the effects of laser stimulation of energetic

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willows. It is recommended to check the calorific value of three-years shoots of
irradiated willows.

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