1 EDUCATIONAL NETWORK ON SOIL AND PLANT ECOLOGY AND MANAGEMENT (EduSapMan) Summer School Soil & Water 2017 Practical exercises Content: 1. Stomatal kinetics in response to CO2 and its relation to stomatal size and density 2 2. Soil compaction and oxygen in soil 12 3. Soil zoology 17 4. Land properties influenced by land use and fertilization 25 5. Fast Plant test with various substrates and composts 49 6. Allelopathy experiment with Estonian trees 60
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1
EDUCATIONAL NETWORK ON SOIL AND PLANT
ECOLOGY AND MANAGEMENT
(EduSapMan)
Summer School Soil & Water 2017
Practical exercises
Content: 1. Stomatal kinetics in response to CO2 and its relation to stomatal size and density 2
2. Soil compaction and oxygen in soil 12
3. Soil zoology 17
4. Land properties influenced by land use and fertilization 25
5. Fast Plant test with various substrates and composts 49
6. Allelopathy experiment with Estonian trees 60
2
1. Stomatal kinetics in response to CO2 and its relation to
stomatal size and density
Markus Hügel, Kateřina Kabeláčová, Thomas Link-Hessing, Claude-Eric Marquet and
Patrick Waldhelm
Supervisors: Tiina Tosens, Linda-Liisa Veromann
1 Introduction
Water use efficiency (WUE) is very often topic lately. The level of greenhouse gas CO2 is
rising. That means, inorganic C is more available for plants to assimilate, but for
photosynthesis is important water also. So on the other hand there is problem with water
sufficiency, because of greenhouse gases, the average temperature of the planet is rising and
dry areas are expanding. Plants are under a constant stress, because of drought.
2 Materials and methods
2.1 Measuring stomatal conductance and photosynthesis
Materials:
- Two plants Platanus orientalis (normal and stressed)
- Fern Microsorum diversifolium
- Two instruments GFS-3000 Portable Photosynthesis System
Procedure:
There were used two plants Platanus orientalis and one fern Microsorum diversifolium. One
P. orientalis was not watered for seven days and the second P. orientalis was well watered.
Plants were kept under artificial conditions. Plane trees were kept under the light intensity of
1000 µmol/m²s, 22 °C and 65% humidity. The M. diversifolium was kept under artificial light
at 600 µmol/m²s, 22 °C and 65% humidity. Experiments were done in the plant physiology
laboratory.
The stomatal conductance and the photosynthesis were measured with two instruments GFS-
3000 Portable Photosynthesis System. At first chambers of the machines were adjusted to
3
these settings: temperature 25 °C and humidity 65%. The light settings differed: For plane
trees it was set to 1000 µmol/m²s and 600 µmol/m²s for M. diversifolium. The CO2 levels in
chambers were changed during measurements. At first the experiment begun with 400 ppm of
CO2, then 100 ppm and at the end 800 ppm. Changes of CO2 levels were made after a
stabilization of assimilation and stomatal conductance. There was also measured the time of a
stabilization.
Figure 1: GFS-3000 Portable Photosynthesis System
Figure 2: P. orientalis leaf in measurement chamber
4
2.2 Stomatal size and density measurement
Materials:
- Transparent nail polish, transparent tape, slides
- Light microscope, PC, Image-J software
Procedure:
The “varnish”-method was used. The leaf which was previously used for measuring, was
coated with clear nail polish on the bottom of the leaf. Leaf-veins were avoided. After the nail
polish was dry, transparent tape was sticked on it and pulled off. The tape with the dry varnish
was sticked on the slides and analysed under the light microscope. Photos of the leaf bottom
imprint were taken in a magnification of 100 and 200. To calculate the stomatal size the
length and width of 10 stomata for each plant were measured with the computer program
ImageJ. The stomatal size was calculated by multiplying the length with the width. To obtain
the stomatal density the amount of stomata on three different parts of the leaf bottom were
counted. To acquire average density the formula: number of stomata / area.
Figure 3: Leaf bottom surface imprint on slides
5
Figure 4: Stomata of M. diversifolium at 200 magnification
6
Figure 5: Stomata of P. orientalis at 200 magnification
3 Results
During the first experiment gained data about the stomatal conductance and the
photosynthesis are in tables 1 and 2. Graphs in figure 6 and 7 were constructed from these
data and from CO2 levels. We calculated values of WUE according to this formula:
𝑊𝑈𝐸𝑝ℎ𝑜𝑡𝑜𝑠𝑖𝑛𝑡𝑒𝑠𝑖𝑠 =𝐴𝑠𝑖𝑚𝑖𝑙𝑎𝑡𝑖𝑜𝑛
𝑠𝑡𝑜𝑚𝑎𝑡𝑎 𝑐𝑜𝑛𝑑𝑢𝑐𝑡𝑎𝑛𝑐𝑒
Table 1: CO2 Assimilation of the three test plants
CO2
[ppm]
P. orientalis (normal) P. orientalis (drought) M. diversifolium
100 1,07 0,02 0,51
400 6,86 0,97 3,5
800 12,26 2,37 6,84
7
Figure 6: CO2 Assimilation of the three test plants
Table 2: Stomata conductance of the three test plants
CO2
[ppm]
P. orientalis (normal) P. orientalis (drought) M. diversifolium
100 103,4 9,1 52,2
400 58,6 9,9 33,7
800 42,2 9,6 33,8
8
Figure 7: Stomata conductance of the three test plants
WUE values were transferred into the table 3 and figure 8. Stabilization periods of plants was
inserted into the table 4.
Table 3: WUE of the three test plants
CO2
[ppm]
P. orientalis (normal) P. orientalis (drought) M. diversifolium
100 10 2 9
400 117 97 103
800 290 246 202
9
Figure 8: WUE of the three test plants
Table 4: Stabilization period of the three test plants
Measurement P. orientalis (normal) P. orientalis (drought) M. diversifolium
1. 40 min 8 min 62 min
2. 13 min 4 min 13 min
3. 22 min 5 min 17 min
Summary 75 min 17 min 92 min
The stomatal size and density are shown in figure 9 and 10.
10
Figure 9: Stomatal size of the three test plants
Figure 10: Stomatal density of the three test plants
11
4 Discussion
We observed reactions of stomata to different levels of CO2. The plant which was stressed by
drought had lower stomata conductance than other plants. It could be related to the
physiological mechanism which tend to protect the plant from complete dryness. The plant
was stressed even more when there was low CO2 level. At the level 100 ppm well-watered
plants managed to open the stomata more than the plant which suffered by drought. At the
level 800 ppm plants had enough of CO2 in the chamber so they could close stomata. Even
though the difference between plane trees is very visible, for better significance and good
strength of the experiment, it would be good to do the experiment with more plants and
different species.
The varnish method showed us how simple it is to observe stomata structures and compare
them between different plant families as gymnosperm and angiosperm. According to the
evolution, we can suggest that angiosperm plants have smaller stomata structure and higher
density compared to ancient gymnosperm family. On the other hand the habitat of the fern is
more humid so it can manage to have bigger stomata than the plane tree. We realized that the
fern needs more time to open stomata and have slower reaction to the changes of CO2 levels.
For better resolution, there would be needed to have wide range of gymnosperm and
angiosperm species to compare.
5 Conclusion
The plane tree which suffered by drought had low stomata conductance during the whole
experiment. The measuring took for 17 minutes in total and gs values were 9,1; 9,9 and 9,6
mmolH2O. WUE values were 2; 97 and 248 µmolCO2∙mmolH2O-1. Well-watered plane tree had
higher values. Stomatal conductance values were 103,4; 58,6 and 42,2 mmolH2O. WUE values
for well-watered plane tree were 10; 117 and 290 µmolCO2∙mmolH2O-1. The measuring took for
75 minutes. The fern had slightly lower values in comparison to well-watered plane tree.
Values of stomatal conductance were 52,2; 33,7 and 33,8 mmolH2O. WUE values were 9; 103
and 202 µmolCO2∙mmolH2O-1. The measuring of the fern took for 92 minutes.
12
Summer School Soil & Water
Tartu 2017
Supervisor: Prof. Dr. Marian Kazda, Ahmed Sharif
2. Soil compaction and oxygen in soil
Linda Ahner
Milan Varsadiya
Laurie-May Gonzales
Bernhard Glocker
Contents 1 Introduction
2 Materials and Methods
3 Results
4 Discussion
5 Conclusion
6 References
13
1 Introduction Nowadays drip irrigation has gained more important in agriculture and has been
used very intensively on the fields. Although this is a more sustained way to
irrigate the plants, it limits oxygen availability for plant roots by creating a nearly
saturated condition. Furthermore, high soil compaction also affects the amount
of oxygen content in soil which changes gas exchange. In general, gas exchange is
also affecting growth and activity of roots and soil organisms, and leading to an
alteration of chemical processes (Ampoorter et al., 2010). For instance, N fertilization has
significant effect on microbial CO2 respiration and communities functioning. This was also
proved by laboratory incubation experiments (Kowalenka et al., 1978).
In Summary soil compaction involves the compression of pores, which leads to
decreased porosity, increase in dry bulk density and reduce hydraulic conductivity.
The questions of this short experiment were “Soil with organic manure has more oxygen
depletion rate than soil without manure” and “Soil managed with organic fertilizer contains
lower bulk density”.
2 Materials and Methods Six samples were collected on 26th of June 2017 from the long-term fertilization
experiment-site IOSDV in Tartu. Two samples were collected from the same field
without organic fertilizers and without manure, two from the same field but with
manure. And the last two samples were collected from the field with an alternative
organic fertilizer.
The samples were collected in metal cylinders from five centimetres under the surface and
these were put in plastic cups. The second part was in the laboratory. First, the samples were
weighted and after that all samples were flooded with double distilled water (ddH2O) until the
complete soil was saturated. All samples got an oxygen sensor through the plastic lid. For the
measurement, the “FIBOX LCD” was used. Periodically, measurements were recorded. When
the reading came to zero, the water from the plastic cups were removed manually.
Then the samples were kept on filter paper and the plastic lids were left half open to test the
speed of re-aeration. Successively the readings were recorded. After all the measurements
have been taken, all wet samples were re-weighted. To measure dry bulk density, the soil has
to be oven-dry. Therefore, samples were kept in an incubator at 105_C for 24 hours. Finally
the dry soils were weighted again.
3 Results In the following Figure 3.1, the results are shown for the changing of the oxygen levels. The
oxygen concentration in all wet soils decreased in the first 17 hours however, the next two
14
days it was stable. On the 1st of July the samples were reaerated at 11 o’clock. The soil without
any fertilizer (beginning value 7.1 mg/l) showed a steep increasing (7.6 mg/l). Then it was on
the same level for a brief time before it drastically declined until 0 mg/l. The Inorganic nitrogen
fertilized sample (beginning value 8.0 mg/l) was stable as well until 1st of July, 16:30. After that
it increased until the end of the experiment (6.2 mg/l). The last sample with organic manure
and nitrogen (beginning value 7.7 mg/l) showed no reaeration until the end (0 mg/l).
Figure 3.1: Changing of the oxygen rate in time. NO – no fertilizer, N 120 –
inorganic nitrogen (120 kg N/ha) fertilizer and ON 120 – alternative
manure nitrogen (120 kg N/ha) fertilizer
Figure 3.2: Dry bulk density. NO – no fertilizer, N 120 – inorganic nitrogen (120
kg N/ha) fertilizer and ON 120 – alternative manure nitrogen (120 kg
N/ha) fertilizer.
Figure 3.2 depicts the dry bulk density (BD). For NO BD the value was 1.4 g/cm3 for the other
two samples the values were smaller. The value for NO 120 was 1.2 g/cm3 and for ON 120
was 1.25 g/cm3.
In Figure 3.3 the maximum water holding capacity is shown. The samples NO, NO 120 and
ON 120 had 30.3 %, 31.3% and 33.3 %, respectively.
15
Figure 3.3: Maximum water holding capacity. NO – no fertilizer, N 120 – inorganic
nitrogen (120 kg N/ha) fertilizer and ON 120 – alternative manure
nitrogen (120 kg N/ha) fertilizer.
The Table 3.1 summarizes variables that are important for soil compaction,
which goes respectably in line together. The less the particle density correlated
with higher porosity and maximum water holding capacity.
Table 3.1: Different soil compaction variables. NO – no fertilizer, N 120 – inorganic
nitrogen (120 kg N/ha) fertilizer and ON 120 – alternative manure
nitrogen (120 kg N/ha) fertilizer.
16
4 Discussion All three samples showed instant decreasing in oxygen concentration suggested
that water created pressure on the air in the soil which lead to escape it from the
soil. This can be proved also in the stable phase afterwards where there cannot
be any significant different seen.
After reaeration, the air can get inside of the soil and provides more oxygen concentration
increased again. Kowalenka et al., (1978) suggested that nitrogen fertilizer has an influential
effect on microbial communities. This can be seen in the results where the oxygen
concentration increased fast in the sample without fertilizer because the microbial activity was
presumably lower. In the sample with just nitrogen fertilizer the microbial activity was better
than in the sample without fertilizer. But only in the sample with nitrogen and organic manure
did not show any changes in the oxygen concentration due to higher microbial activity.
However, there is still a need to optimize the method as there were problems in enclosing air
bubbles in front of the sensor. Furthermore, moving the sensors during the measurement could
cause its shift into soil parts still anoxic (c.f. Fig. 3.1., sample NO).
The bulk density and the maximum water holding capacity showed a negative
relationship. The higher the bulk density, the less the porosity which lead to less
water holding capacity. The particle density correlated with the porosity.
5 Conclusion Irrespectively the soil properties, in flooded soils the water replaces the air in the
pores so there is nearly no oxygen left. There wasn’t any substantial change in
reaeration for organic manure with mineral N fertilized soil. Its most likely due
to the microbial activities in the soil Addicted to mineral nitrogen fertilizer with
manure, there is a lower bulk density and a higher water holding capacity. , which
depends on soil composition.
In the end, the first hypothesis could not be proved in the depletion rate but
there was no increase in the oxygen rate in the soil managed with manure while
reaeration. Just like the first hypothesis the second was not able to be proved: the
bulk density was not lower in soil with the organic fertilizer.
6 References Ampoorter, E., Van Nevel, L., De Vos, B., Hermyc, M., Verheyena, K. 2010.
“Assessing the effects of initial soil characteristics, machine mass and traffic
intensity on forest soil compaction”. Forest Ecology and Management 260,
1664–1676.
Kowalenko, C. G., Ivarson, K. C., Cameron, D. R. 1978. “Effect of moisture
content, temperature and nitrogen fertilization on carbon dioxide evolution
from field soils”. Soil Biology and Biochemistry 10, 417-423.
Bhattarai, S. P., Pendergast, L., Midmore, D. J. 2006, “root aeration improves
yield and water use efficiency of tomato in heavy clay and saline
Table 1 shows the entire results of the experiments. All following results will refer to this table.
Table 1 All measured values results
P (mg/kg) Mg (mg/kg) Ca (mg/kg) N (%) C (%) pH
Grasland 1 122 130 973 0,17 2,23 5,00
Grasland 2 108 131 789 0,16 2,15 4,73
Grasland 3 102 147 1 006 0,17 2,12 4,84
N0/1 133 149 1 332 0,08 1,25 6,39
N0/2 128 146 1 258 0,06 1,14 6,21
N0/3 135 163 1 360 0,07 1,17 6,23
N120/1 126 112 787 0,07 1,01 5,38
N120/2 124 90 738 0,07 0,99 5,11
N120/3 161 134 1 119 0,07 0,96 5,38
N0 org fert/1 136 167 1 294 0,07 1,10 6,28
N0 org fert/2 143 147 1 110 0,07 1,19 6,25
N0 org fert/3 141 163 1 336 0,08 1,17 6,18
N120 org fert/1 148 121 1 036 0,08 1,11 5,85
N120 org fert/2 154 152 1 148 0,07 1,11 6,09
N120 org fert/3 123 131 892 0,09 1,09 6,09
3.1 Soil texture fingering test
Conductivity
(µS) Moisture (%)
Bulk density
(g/cm3)
Calc. Bulk
density
(g/cm3)
SOC stock
(t/ha)
Grasland 1 58 1,19 1,51 50,67
Grasland 2 45 1,51 1,51 48,71
Grasland 3 43 1,60 1,50 47,66
N0/1 70 1,10 1,61 1,55 29,05
N0/2 67 1,00 1,54 1,55 26,57
N0/3 63 0,90 1,56 27,39
N120/1 87 1,10 1,44 1,57 23,83
N120/2 39 1,10 1,34 1,57 23,25
N120/3 98 -19,00 1,58 22,81
N0 org fert/1 32 1,00 1,56 25,72
N0 org fert/2 60 0,90 1,57 28,00
N0 org fert/3 27 1,01 1,56 27,38
N120 org fert/1 69 0,90 1,43 1,58 26,37
N120 org fert/2 75 1,00 1,54 1,56 26,03
N120 org fert/3 90 1,10 1,58 25,80
34
The soil texture type was determined to
be sandy loam with clay content of 10 –
25% according Food and agriculture
organization of the United Nations
(2006) (Fig. 2). The result was equal for
all the samples.
3.2 Moisture content by air drying
The measured values are shown in Table 1. They are generally between 0.9 and 1.6 %. The
lowest value showed moisture of -19 %.
3.3 Soil pHKCl
The pH of the grassland soil is 4.86. The field without any fertilizers used, N0, is 6.28 which is
also the highest measured pH. The N0 organic is treated with organic fertilizers and has a pH
of 6.24. N120 has a pH of 5.29 and the pH of N120 organic is 6.01 (Fig. 3).
Figure 4 Comparison of sample taken for fingering
test with the guideline picture
35
Figure 5 Soil pHKCl depending on land use and fertilisation. N0: without organic fertilizer and without mineral nitrogen, N0 organic: with orgnic fertilizer every third year (manure) and without mineral nitrogen, N120: without orgnic fertilizer and with 120 kg N/ha, N120 organic: with orgnic fertilizer every third year (manure) and with 120 kg N/ha
3.4 Soil electrical conductivity/salinity
The Figure 4 and the Table 1 show the soil electrical conductivity of the soil solution of the
different samples. The N0 organic has the lowest soil electrical conductivity with 39.67 µS and
the N120 the highest with 74.67 µS.
Figure 6 Electric conductivity of soil solution depending on land use and fertilisation. N0: without organic fertilizer and without mineral nitrogen, N0 organic: with orgnic fertilizer every third year (manure) and without mineral nitrogen, N120: without orgnic fertilizer and with 120 kg N/ha, N120 organic: with orgnic fertilizer every third year (manure) and with 120 kg N/ha
4,86
6,28 6,24
5,29
6,01
-
1,00
2,00
3,00
4,00
5,00
6,00
7,00
Grassland N0 N0 organic N120 N120 organic
pH
48,67
66,67
39,67
74,67 78,00
-
20,00
40,00
60,00
80,00
100,00
120,00
Grassland N0 N0 organic N120 N120 organic
µS
36
3.5 P, K, Ca, Mg by Mehlich-3 method
Figure 5 represent the measured amount of macro-nutrients in different fertilized crops, no
fertilized crop and grassland.
The received data was compared with reference values and it turned out that phosphorus has a
high amount in each case. In-between the different crops there have been no big differences in
the Phosphorus amount, what can be seen in figure 1. Just the grassland soil has a little bit less
phosphorus than the arable soils, with approximate 110 mg/kg. The highest amount with
approximate 145 mg/kg phosphorus can be found in the soil of N120, which has been treated
with organic fertilizer.
For magnesium, there is a medium content to the reference values and a high content in the zero
fertilized crops, plus in the crops which has been treated with organic fertilizer. The highest
amount is almost 160 mg/kg Magnesium in the NO crops with organic fertilizer, and the lowest
amount can be found in the N 120 crop with approximate 110 mg/kg.
Potassium reached a higher amount with over 180 mg/kg in grassland, than in the fertilized
crops. The lowest amount is shown in the N 120 crop with less than 60 mg/kg of potassium.
Figure 7 Measured P, K, Mg by Mehlich-3 method depending on land use and fertilization. N0:
without organic fertilizer and without mineral nitrogen, N0 organic: with orgnic fertilizer every
third year (manure) and without mineral nitrogen, N120: without orgnic fertilizer and with 120
kg N/ha, N120 organic: with orgnic fertilizer every third year (manure) and with 120 kg N/ha
37
Figure 6 shows the measured Calcium amount by the Mehlich-3 method in each arable soil.
There is the highest amount of Calcium in the no fertilized crop (NO) with approximate 1350
mg/kg Calcium. The lowest amount can be found in the crop which has been treated with
mineral fertilizer (N 120). It indicates an amount of approximate 900 mg/kg Calcium.
There is a positive correlation between the pH and the phosphorus, what can be seen on figure
7.
0
200
400
600
800
1000
1200
1400
Grassland N 0 N 120 N 0 organic N 120 organic
mg/
kg
Ca, mg/kg
0
20
40
60
80
100
120
140
160
0
1
2
3
4
5
6
7
Grassland N 0 N 120 N 0 organic N 120 organic
mg/
kg
pH pH
P
Figure 8 Measured P, K, Mg by Mehlich-3 method depending on land use and fertilization
Figure 9 Phosphorus correlated with pH depending on land use and fertilization
38
3.6 Loss on ignition (LOI)
The results of loss on ignition show the highest loss in permanent grassland and very similar
values in the other fields while the lowest values are measured in the field fertilized with
nitrogen (Fig. 8).
Figure 10 Average loss on ignition of different experimental field managements.
0
0,5
1
1,5
2
2,5
3
Grassland N 0 N 120 N 0 organic N 120 organic
LOI
%
Average Loss-on-ignition
39
3.7 C:N by dry combustion
The results show strong positive correlation between C and N percentage in soil samples. The
result which is far from the others is the grassland with significantly higher percentage of both
carbon and nitrogen (Fig. 9).
Figure 11 C:N Correlation in soil samples from different management of field and grassland measured by Dumas dry combustion.
y = 0,086x - 0,0215R² = 0,9685
0
0,02
0,04
0,06
0,08
0,1
0,12
0,14
0,16
0,18
0 0,5 1 1,5 2 2,5
N in
%
C in %
40
3.8 Dry bulk density
Table 2 is showing the calculated dry bulk density with the equation _ and the average.
Table 2 Measured dry bulk density and calculated average
The average was used to create the graph in Figure 10. In Figure 10 the dry bulk density
depending on land use and fertilization is shown. The difference between the maximum at N0
and minimum at N 120 is 0.18 g cm-3.
Figure 12 Measured dry bulk density depending on land use and fertilization
-
0,20
0,40
0,60
0,80
1,00
1,20
1,40
1,60
1,80
N 0 N 120 N 120 organic
g/cm
³
dry bulk density
bulk density
Sample Bulk density Bulk density average
N0 1,61
1,57 N0 1,54
N 120 1,44
1,39 N120 1,34
N 120 organic 1,43
1,48 N120 organic 1,54
41
3.9 Calculated soil carbon stock
Because of the little amount of values, the bulk density was calculated with a formula which is
shown in the material and method part. The grassland too, because then the different values can
be compared.
The calculated average of the soil organic carbon stock depending on land use and fertilization
is shown in Figure 11. The soil organic carbon stock is indicated in t h-1. There is more carbon
stock in the grassland and less in the agricultural fields. Between the agricultural fields is no
mentionable difference of the soil organic carbon stock.
Figure 13 Calculated soil carbon stock depending on land use and fertilization
0
10
20
30
40
50
60
Grassland N0 N120 N0 organic N120 organic
SOC
in t
/ha
Soil organic carbon
42
3.10 Correlation matrix
The correlation matrix shows strong correlation between all measurements connected to soil
carbon. There is quite strong correlation between carbon and nitrogen and also between nitrogen
and phosphorus. There is quite strong correlation between phosphorus available and pH too
(Tab. 3).
Table 3 Correlation matrix of all collected data
P, mg/kg Mg, mg/kg K, mg/kg Ca, mg/kg N%
P, mg/kg 1,00
Mg, mg/kg 0,22 1,00
K, mg/kg - 0,56 0,25 1,00
Ca, mg/kg 0,47 0,86 - 0,05 1,00
N% - 0,69 - 0,08 0,90 - 0,38 1,00
C% - 0,68 0,03 0,92 - 0,26 0,97
pH 0,56 0,59 - 0,54 0,75 - 0,73
µS 0,37 - 0,23 - 0,42 - 0,13 - 0,29
% water content in soil sample - 0,52 0,05 0,17 - 0,07 0,19
LOSS ON IGNITION - 0,75 0,11 0,92 - 0,16 0,90
bulk density 0,27 0,94 0,82 0,91 0,04
C% pH µS
% water content in soil sample
LOSS ON IGNITION
bulk density
P, mg/kg
Mg, mg/kg
K, mg/kg
Ca, mg/kg
N%
C% 1,00
pH - 0,65 1,00
µS - 0,35 0,11 1,00
% water content in soil sample 0,25 0,13 - 0,48
1,00
LOSS ON IGNITION 0,95 - 0,60 - 0,47
0,31 1,00
bulk density 0,88 0,93 0,51
0,00 0,70
1,00
43
4. Discussion
4.1 Soil texture by fingering test
The result of the soil texture was similar on the field as on the grassland because soil texture
doesn’t change so quickly, thus no changes can be observed after 30 years of experimental
agricultural use of this particular site. This result was expected and was described in the
guideline of the experimental site (Astover Alar, Estonian University of Life Science, 2017).
4.2 Moisture content by air drying
The moisture of the soil according the numbers is quite low comparing to climate conditions in
Estonia, where the precipitation is higher than evaporation. All the samples showed quite the
same humidity and didn’t show any significant difference between various management types.
The lowest humidity value shows unrealistic result and was probably caused by typing error.
4.3 Soil pHKCl
There are differences between the pH of the grassland and the pH of the fields. The moderate
acid pH of the grassland is as expected because of the natural process of acidification and no
presence of carbonates in the soil. The pH of the field-samples is higher because of the liming
made in year 2000 on the field. If there are no fertilizers added as in the Plot N0 there is no
anthropogenic influence on the pH. The higher amount of mineral fertilizers in N120 causes a
lower pH than the N0, because the bacteria oxidase the ammonium and they release hydrogen-
ion during this process. Organic fertilizers contain calcium and magnesium which neutralizes
the pH because they improve soil buffering-capacity. This also could be a reason for the higher
pH than in the grassland.
4.4 Soil electrical conductivity/salinity
The general trend shown in Figure 3 is that the more fertilizer is used, the higher is the soil
electrical conductivity of our samples. That is totally what was expected because when there
are some mineral fertilizers added, ions responsible for electrical conductivity are added the
44
same time. The N0 organic sample doesn’t fit in the trend because the soil electrical
conductivity is lower than the N0 without any fertilizer used. The high standard deviation of
N0 organic shows that the measured result is not very representative. Also the outer standard
deviations are high, because of the fluctuations of the values shown by the instrument while
measuring.
4.5 P, K, Ca, Mg by Mehlich-3 method
The phosphorus concentration is quite high in all arable soils, because it has been fertilized with
phosphorus 20 years ago, so an amount can still be found.
As the phosphorus assimilation benefits by the mutualism with mycorrhiza, a lower amount of
phosphorus in soil could be expected for grasslands. This might due to a higher density of intact
root systems in grassland, so more phosphorus is taken out by the plants. Moreover, the
grasslands haven´t been fertilized, why the phosphorus source in the soil decreases over the
years. Phosphorus is an important primary macronutrient for plants to build the DNA and to
guarantee the membrane development and function. For that reason, fertilization is essential to
assure a good yield.
The higher amount of potassium in grassland can be explained by the absent harvest. Thus,
there has been no potassium taken out of the grassland over the years. In contrast to that, on the
fertilized crops the potatoes have a high demand of potassium, which is taken out more and
more by every harvest.
In acidic conditions phosphorus is more available in a free mineral form, thus it can be used by
microorganism. When the pH is to low ore to high, phosphorus is fixed for example with iron
as a complex, for which reason it is not available any more.
4.6 Loss on ignition (LOI)
The highest loss on ignition measured in grassland soil sample shows the highest amount of
soil organic matter. The management of mowing only enables soil organic carbon to accumulate
in the upper soil layers. Although the other results differences are slightly insignificant, the
lowest carbon content should be observed in the nitrogen only fertilized fields because nitrogen
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addition increase decomposition by saturating the decomposers’ need for nitrogen. The organic
only fertilized field shows relatively high loss on ignition because organic manure contents high
amount of organic carbon which can be quite stable in the soil.
4.7 C:N by dry combustion
The strong correlation between C and N amount shows that nitrogen in soil is mainly bound in
organic particles.
4.8 Dry bulk density
The values are between 1.39 g cm-3 and 1.57 g cm-3 which is typically, because the density of
mineral soils commonly ranges from 1.1 to 1.5 g cm-3 in surface horizons [1]. The little
deviations can come from samples, which were taken with compaction or crumbling.
The difference between the maximum and minimum from the fields with 0.18 g cm-3 is very
low. The reason for this is that the used samples are from the same field with the same soil. It’s
no surprise because the soil changes slowly in the landscape. Maybe there would be a difference
when grassland samples are compared. In fact of this, in the grassland should be more carbon
than in the other samples.
4.9 Calculated soil carbon stock
There is less soil organic carbon stock in the agricultural fields because plants are taking carbon
from the atmospheric CO2. The SOC in the grassland is much higher because there are no plants
which use a high amount of carbon from the atmospheric CO2 In fact of this the amount of
carbon in the soil is higher. Between the different agricultural fields isn’t a significant
difference. It doesn’t matter which fertilization, the organic carbon left in the soil is always
quite the same.
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4.10 Correlation matrix
The correlation between organic carbon related measurements (loss on ignition, carbon by dry
combustion, bulk density, calculated bulk density, total carbon stock) is obvious. The
correlation between C:N:P shows that the main source of active N and P in soil is the soil
organic matter which contains all these elements.
The correlation between pH and Phosphorus shows that soil phosphorus is available in very pH
neutral conditions only because too high (to low) pH cause binding of P into unsolvable
complexes. This is why in grassland (where pH is low) the P available is low too.
4.11 Comparison
The results of this study are comparable with the results of other analyses on the effect of
mineral and organic fertilization on soil. Körschens et al. [2] compared the results of 20
European long-term experiments concerning the impact of fertilization on crop yield, carbon
balance, soil organic carbon content and dynamics. In Figure 12, their work is compared to the
work at hand. The figure demonstrates the effect of fertilization and clay content on the soil
organic carbon content for 18 European long-term experiments and the results for Tartu. From
the left to the right side, the clay content of the sites is increasing. The lighter part of the columns
shows the soil organic carbon content without fertilization and the darker part shows the content
with organic (10 t/ha FYM) and mineral (NPK) fertilization. The results concerning the clay
content and the carbon content for Tartu are added in red. The red frame contains the range of
the clay content for Tartu according to the fingering test.
47
Figure 14: Comparison of our measurements with the result of Körschens et al. [1]
For Tartu, we measured a clay content of 10-25 %. In comparison, Speyer has a clay content of
9 % and Wien a clay content of 25 % [2]. For the treatment without fertilization, Tartu got a
result of 1.19 % of carbon, Speyer 0.58 % and Wien 2.06 % of SOC. So the result of Tartu lies
in between the results of Speyer and Wien, what is consistent with the expectations. The result
for the treatment with mineral and organic fertilizer for Tartu was 1.10 % whereas the result for
Speyer was 0.81 % and for Wien 2.24 % SOC. Again, the value of Tartu lies in between the
values of Speyer and Wien. However, according to [2], for Speyer and Wien, the carbon content
for the treatment with fertilization was higher than for the treatment without fertilization while
for Tartu, it was the other way round. To summarize, the values for Tartu are in accordance
with the results by Körschens et al. even if the fact that for Tartu the carbon content decreased
while using organic and mineral fertilizer is surprising.
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5. Sources
[1] Thien and Graveel, Laboratory Manual for Soil Science
[2] Martin Körschens et al. (2012) Effect of mineral and organic fertilization on crop yield,
nitrogen uptake, carbon and nitrogen balances, as well as soil organic carbon content and
dynamics: results from 20 European long-term field experiments of the twenty-first century.
Archives of Agronomy and Soil Science, 59:8, 1017-1040
49
5. Fast Plant test with various
substrates and composts.
PROTOCOL By Laura Böe, Aneta Flekalova, Donatus Mmodum, Arthur
Naimowitsch.
OUTLINE
1. Introduction
2. Material and Methods
3. Results
4. Discussion
5. Future perspectives
1. Introduction
Worldwide peat is decreasing especially in Europe. This made us work on the alternative for this and
Compost could be the alternative for this peat growing media therefore Compost is good for
Agriculture because of its resource usage efficiency especially in Nutrients and Organic matter. This is
an important project because it centers on the possible replacement of peat growing media if we
eventually lose our peat especially in Europe. In this our study we compare and test different quality
of composts by using fast growing plant tests to find out the difference on compost quality. We try to
50
find out if different quality of compost from different source can cause growth in Plants. And also if
the compost quality determines the growth in plant. Below is the distribution of peat in Europe.
PEAT DISTRIBUTION IN EUROPE
This paper derives the distribution of peat land in Europe as the extent of peat and peat-topped soils
indicated by soil databases. The data sources were the 1:1,000,000 European Soil Database (v1.0)
and a data set of organic carbon content (%) for the top soils of Europe at 1km x 1km resolution that
was recently published in map form.
The strong influences of vegetation and land use on soil organic carbon (OC) content were taken
into account in computing the 1km (OC) data set, as was the influence of temperature.
The areas of peat and peat-topped soils estimated from the European Soil Database are generally in
close agreement with those obtained using the Map of OC in Top soils of Europe. The results reveal a
strong northern bias in the distribution of organic soils across Europe. Almost one-third of the peat
land resource of Europe is in Finland, and more than a quarter is in Sweden. The remainder is in Poland,
the UK, Norway, Germany, Ireland, Estonia, Latvia, The Netherlands and France. Small areas of peat
and peat-topped soils also occur in Lithuania, Hungary, Denmark and the Czech Republic. For most
European countries, the distribution of peat and peat-topped soils is probably more accurately
portrayed by the Map of OC in Top soils of Europe than by the European Soil Map and Database. Such
baseline data are important for the conservation of peat and for making much more precise estimates
of carbon stocks in topsoil than have been possible hitherto.
The results are also relevant to the planning of effective soil protection measures at European level
Allelopathy is a chemical interaction between plants, and between plants and microorganisms. Plants produce allelochemicals (secondary metabolites), which are released into the environment. These secondary metabolites have influence on growth, germination, reproduction, distribution of vegetation.
In our experiment, we have been observing allelopathy of 3 common Estonian plant species: Acer platanoides, Picea abies and Quercus robur. From scientific articles, we know that allelopathy of boreal shrubs has been evidenced, but we have low knowledge about allelopathy of Estonian trees (Gallet 1994).
We raised 2 main hypotheses:
- H1: Macerate of leaves or needles of Estonian common trees (Acer platanoides, Picea abies, Quercus robur) have allelopathic effect on germination of Lactuca sativa, Lepidium sativum.
- H2: allelopathic effect of young needles of Picea abies is higher compared to old needles of Picea abies.
Our experiment was mainly done by germination tests.
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Material and method
1. First experimentation
The first experimentation is carried out to observe the potential allelopathic effect of the macerates
of needles of Picea abies and of leaves of Acer platanoides.
i. Preparation of macerates
We put 20g (fresh weight) of needles of Picea abies or leaves of Acer platanoides in a glass with
100ml of distilled water. After 17 hours we filtered the macerate, this was the mother solution at
10% (mass/volume). Another solution of 5% was realized by mixing 50ml of the solution at 10% with
50ml of distilled water.
A control was also realized with only distilled water (0%).
6 treatments were tested: 2 species (P. abies and A. platanoides) at 3 concentrations (5%, 10% and
control at 0%).
ii. Germination test
For the germination test we used Petri dishes with a substrate of filter paper. In this filter paper, we
put 25 seeds of the target species: Lepidium sativum.
There were 24 Petri dishes: 6 treatments and 4 replicates per treatment.
We watered each Petri dish with 2ml of one solution (distilled water as 0% control, 5% or 10% of
macerate of each species).
48 hours after watered the Petri dishes, we counted the number of germinated seeds, and the
germination stage, according to the Figure 1.”.
Figure 1: the 4 germination stages considered for the experiment. 1: no germination, 2: germinated
without leaves, 3: germinated with yellow leaves, 4: germinated with green leaves.
Data analysis was done with Chi² tests.
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2. Second experiment
The first experiment demonstrated no significant inhibition effect of the tested treatments. We
performed a second experiment, with the same protocol but we used 2 target species (Lepidium
sativum and Lactuca sativa), and we selected two tree species Picea abies and Quercus robur and
two phenological stages for Picea abies (young and old needles).
i. Preparation of macerate
We put 20g (fresh weight) of leaves or needles of Quercus robur and young or old needles of Picea
abies in a glass with 100ml of distilled water. After 17 hours, we filtered the macerates.
A control for the macerate is also realized with only distilled water (0%).
8 treatments were tested: 3 macerates (young or old needles of Picea abies or leaves of Quercus
robur) at 10% and 1 control (0%), with 2 target species (Lepidium sativum and Lactuca sativa).
ii. Germination test
For the germination test we used Petri dishes with a substrate of filter paper. In this filter paper, we
put 25 seeds of the target species: Lepidium sativum and Lactuca sativa.
There were 24 Petri dishes: 8 treatments and 3 replicates per treatment.
We watered each Petri dish with 2ml of one solution (distilled water as control or macerate).
48 hours after watered the Petri dishes, we counted the number of germinated seed, and the
germination stage, according to the Figure 1.
Data analysis was done by using Chi² tests.
Table 1: Summary table of both experiments:
First experimentation Second experimentation
Number of 0%
control (distilled
water)
8 3
Donor plants 2 (Picea abies and Acer
platanoides)
3 (young and old needles of Picea
abies and leaves of Quercus robur)
Concentrations 5% and 10% 10%
Target species 1 (Lepidium sativum) 2 (Lepidium sativum and Lactuca
sativa)
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Replicates 4 3
Results
1. First experiment
Figure 2: Germination rates of Lepidium sativum with 0% control with distilled water, with 5 and 10%
macerates for the two donor plant species (C=conifer=Picea abies, D=deciduous=Acer platanoides).
Results showed that most of the seeds germinated (Figure 2). All treatments lead to all the
germination stages, except D5 (Acer 5%), which had no stage 0 (all seeds have germinated). D10
(Acer 10%) treatment showed the highest proportion of non-germinated seeds. Both Picea