Effects of Different Additives on Soil Respiration and

Nitrogen Transformations (Lab 15)
By: Tyler-Anne Buck
Soil Science: Fall Semester 2015
Abstract:
The purpose of this experiment was to assess the effects that different amendments have
on soil respiration and nitrogen transformations. Over 3 weeks, jars containing the same soil,
water and different soil amendments were incubated and then titrated to extract the amount of
carbon dioxide that was released. The amount of nitrate and ammonium were tested using test
strips and drop kits respectively. The soil with added alfalfa measured the highest average in all 3
of the tests. This suggested that alfalfa promoted soil respiration and the transformation of
nitrogen via ammonification and nitrification.
Introduction:
Soil respiration is very important in understanding the global carbon cycle. It can be
influenced by a variety of factors such as temperature, moisture content, relief, and organisms.
Soil respiration can indicate a general trend in the rate of reactions and activity of decomposers.
The more respiration that is occurring, the more carbon dioxide that is being released. In places
such as the tropics, the abundant plant life and conditions are ideal for decomposers and so
respiration and, therefore, carbon dioxide release is high. The soil, however, can be a great
carbon sink where there are low respiration rates. (Schlesinger et al. 2000)
There are many other nutrient cycles that occur other than just the carbon cycle. The next
most involved cycle is that of nitrogen. Plants use nitrogen for a variety of things such as for
proteins, chlorophyll and DNA but they can only uptake nitrogen in a soluble form through

ammonium or nitrate. Decomposers and bacteria make organic nitrogen available. This organic
nitrogen then can become ammonium through ammonification which can further become nitrate
through nitrification. Generally, sawdust has a carbon to nitrogen (C/N) ratio of 600:1 and alfalfa
has a CN ratio of 13:1. Soil organisms need 1 g of nitrogen for every 24 g of carbon that they
consume and so any soil that has a ratio lower than 24:1 will have a net mineralization of
nitrogen. On the other hand, in soils with a higher than a 24:1 ratio, decomposers will take
soluble nitrogen from the soil solution and decrease the availability of nitrogen available to
plants which is found in forms of ammonium and nitrate. (Brady et al., 2010) This also slows
decomposition. Based on the C/N ratios of sawdust and alfalfa, it is hypothesized that the alfalfa
jar will respire more and contain higher amounts of nitrate and ammonium than the sawdust
because more nitrogen is available for processes such as ammonification and nitrification. The
Methods:
This experiment took place over multiple weeks. 4 mason jars were incubated for 3
weeks. Before the incubation period, 3 jars received 50 grams of garden soil each. One of those
jars received 0.5g of alfalfa, the second jar received 0.5g of sawdust, and the third jar received no
additional amendment and was labeled the “control” jar. 5mL of distilled water was added and
mixed into to the soil. The fourth jar received no water or soil and was labeled the “blank jar”
and was used as a baseline for comparison. Test tubes containing 20mL of 2M sodium hydroxide
(NaOH) were placed in each of the 4 jars. These jars were then sealed and left to sit for the three
weeks, only being opened intermittently to allow the oxygen to be replenished.
At the end of the incubation period, the NaOH in each solution was titrated using 1M
BaCl2, a Phenolphthalein, and hydrochloric acid (HCl), in order to determine how much NaOH
was neutralized from the carbonic acid (H2CO3). The H2CO3 formed from the carbon dioxide

(CO2) being released from soil respiration and dissolving in the NaOH. The amount of CO2 lost
was then calculated. Next, the soil itself was mixed with water and then filtered. The filtered
solutions were each tested for nitrate (NO3-) using nitrate test strips and for ammonium (NH4+)
using an ammonium test kit. Lastly, a t-test was used to analyze the significance between the
amendments for soil respiration and for nitrate.
Results:
An addition of alfalfa produced the highest amount of CO2, nitrate concentration and
ammonium concentration. The Mason jar with the alfalfa additive supported the most amount of
soil respiration with an average of 0.081g of CO2. The sawdust amended jar had the second
highest amount of CO2 released, with an average of 0.035g. The control jar had the least amount
of CO2 released with an average of 0.002g (See Figure 1). The measured amounts of nitrate
followed the same trend. The alfalfa jar averaged 2.33 ppm of NO3-, the sawdust jar averaged 1.8
ppm of NO3-, and the control jar average 1.75 ppm of NO3- ( See Figure 2). The results of the
measured ammonium differed slightly. The alfalfa jar had the highest average NH4+ with 52.5
ppm. The second highest was that of the control jar with an average of 2ppm. The sawdust jar
had the least amount with an NH4+ average of 0 ppm (See Figure 3). The t-test for soil respiration
comparing the alfalfa to the sawdust produced a p-value of 0.135 (13.5%). The t-test for nitrate
comparing the sawdust to the alfalfa produced a p-value if 0.613 (61.3%)
Discussion:
The results support the hypothesis that the low C/N ratio in the alfalfa supported the
production of more nitrate and ammonium. It also produced a higher amount of CO2 which
implies that it had a higher rate of soil respiration. Sainju et al., 2010 found that the use of alfalfa

increased the amount of nitrogen stored in the soil and increased mineralization of it. This further
agrees with the low C/N ratio hypothesis in which more nitrogen is available. In comparison with
the control, both the alfalfa jar and the sawdust jar had elevated amounts of soil respiration and
nitrate but sawdust showed no ammonium concentrations. The p-values (61.3% and 13.5%
respectively) from the both soil nitrate and the soil respiration t-tests implies that the mean
nitrate and CO2 produced were significantly different.
In a study by Iannone et al., 2009, nitrate concentrations in soils with added sawdust
resulted in depleted amounts of nitrate and no depletion of ammonium. Theoretically, nitrate
levels in the sawdust jar could be lower due to the nitrate being used up through immobilization,
plant uptake, or denitrification. This dissimilarity could have resulted from low oxygen levels
inhibiting the immobilization and uptake due to the jars not being aerated often enough.
These findings on alfalfa are in concurrence with findings in other studies as to how and
why alfalfa is a strong soil additive. In a study by Qian et al., 2011, they discuss that alfalfa, in
moderation, is an effective additive as it supplies more nutrients to plants, especially over short
periods of time without strongly affecting other properties such a pH. This can aid farmers in
increasing their crop yield. In addition, the activeness of decomposers and nitrogen fixing
bacteria can have hefty influences on soil respiration, and therefore the amount of the greenhouse
gas of CO2 that is released into the atmosphere.
Conclusion:
High soil respiration, nitrate levels and ammonium levels developed from the addition of
the alfalfa, which had a low C/N ratio. This ratio implies that more nitrogen was available for
development of nitrate and ammonium. The soil with added sawdust, which had a higher C/N

ratio, did not respire as much, had similar nitrate levels to the control jar, and produced no
ammonium. This reveals that the general amounts of nutrients, such as that of carbon and
nitrogen, play an important role in the interaction between the plants and the soil. Soil respiration
due to decomposers and nitrogen fixing bacteria have a large influence on the carbon cycle as
well. Repeated studies using this method with timely set intervals for aeration of the jars may be
able to determine whether or not the high nitrate levels in the sawdust was due to the lack of
oxygen.

References:
Brady, N. C., & R. R. Weil, 2010. Elements of the Nature and Properties of Soil (3rd ed.). Upper
Saddle River, New Jersey.
Iannone, B. V., C. J. Rosen, & S. M. Galatowitsch, 2009. Soil Nitrogen Concentrations in a
Restored Sedge Meadow Wetland as Affected by the Application of High C:N
Amendments. Ecological Restoration, 193-199.
Qian, P., J. J Schoenau, & C. Fatteicher, 2011. Effects of Soil Amendment with Alfalfa Powders
and Distillers Grains on Nutrition and Growth of Canola. Journal of Plant Nutrition,
1403-1417.
Sainju, U., & A. W. Lenssen, 2010. Soil Nitrogen Dynamics under Dryland Alfalfa and DurumForage Cropping Sequences. Soil Science Society of America Journal, 669-677.
Schlesinger, W. H., & J. A. Andrews, 2000. Soil Respiration and the Global Carbon Cycle.
Biocheochemistry, 7-20.

Figures/Tables:

g CO2
0.09
0.08
0.07
0.06
0.05
0.04
0.03
0.02
0.01
0

Control

Alfalfa

Sawdust

Figure 1: This graph shows the average amount of CO2, in grams, and the standard error calculated in each jar.

ppm NO32.5
2
1.5
1
0.5
0

Control

Alfalfa

Sawdust

Figure 2: This graph shows the average amount of nitrate in each jar along with the standard error.

ppm NH4+
60
50
40
30
20
10
0

Control

Alfalfa

Sawdust

Figure 3: This graph shows the average amount of ammonium measured in each jar.