Discussion

Many different properties of forest ecosystems were investigated to determine the effect

that long-term nitrogen addition would have on them. The first of these properties was the Net

Primary Productivity of the trees in these two types of forests (hardwood and pine), and the

results of this experiment were interpreted by analyzing a scatter plot and its connecting lines,

and thus determining the trend that exists between nitrogen levels over time and the productivity

of these trees. By definition, the net primary productivity of a plant or plants is “the rate of

storage of organic matter in plant tissues in excess of the respiratory utilization by plants." (Sala

and Austin 2000). Keeping this in mind, it can be concluded from the graphs that with an

increased level of nitrogen in the surroundings of these trees, there is a notable increase in the

primary productivity of hardwood trees, but a decrease in that of pine trees, over the given time

frame. In the case of hardwood trees, the primary consumers that would ingest these plants

would see a greater benefit from them, due to this increased storage of nutrients. The forest

ecosystem, therefore, would flourish. This positive correlation could be caused by the

importance of nitrogen in the light independent reactions, also known as the Calvin Cycle, of

photosynthesis (Evans 1989). Additionally, the chlorophyll present in the thylakoid membranes

is heavily dependent on nitrogen in order to maintain their functionality, and with increasing

nitrogen levels, the light dependent reactions are also affected. However, with the pine trees,

the above does not apply. Initially, one could assume that the pine stand would exhibit similar

results as the hardwood stands in relation to primary productivity, since the nitrogen

concentration should improve the process of photosynthesis, but the data does not support this

claim. According to the scatter plot, an increased level of nitrogen addition causes the primary

productivity of pine trees to decrease. An assumption can be made that this difference in trend

could be caused by differing light requirements for hardwood and pine trees, differing oxygen

levels or types of soil required for optimum primary productivity. It is also important to note that
different species of microorganisms could inhabit the roots of pine and hardwood trees, and

these microorganisms could differ in their output of nitrogen i.e nitrogen fixation depending on

the types of nutrients they would require.

The next point of analysis is the relationship between the photosynthetic capacity and

Nitrogen concentration over time. Photosynthesis refers to the process by which plants convert

energy from sunlight into chemical energy and releases oxygen into the atmosphere.

This can be simplified by the equation: 6 H2O + 6 CO2 + energy → C6H12O6 + 6 O2 (Farquhar et al.

2001). Capacity is a measure of the maximum rate of the reaction and evaluates the “efficiency”

of a plant depending on different factors. Here, it is about looking for a correlation between the

amount of nitrogen in a plant and how much photosynthesis can occur, which can be defined by

the amount of carbon dioxide consumed (in nmol/gs). This quantity is measured for two types of

tree samples, one with a control (normal) level of N2 and the other with a higher concentration,

annually for 4 years. The results were then organized into a bar graph which compares the two

samples every year and includes the errors for each measurement. As was discussed above,

nitrogen has an impact on the photosynthetic abilities of plants due to its integral structural role

in plant proteins (Evans 1989). Although the aspect of the graph suggests that higher levels of

nitrogen reduces photosynthetic capacity, the results remain in possible disagreement so no

conclusion can be made. Therefore, the experiment shows no significant relationship between

photosynthetic capacity and nitrogen concentration. Taking into consideration that human

activity is responsible for a rise in nitrogen levels, it seems that mankind is not a threat to

oxygen levels and plant productivity. However, the uncertainty in the results as well as other

factors which reduce air quality (greenhouse gasses) support the conclusion that human

activities have a negative effect on the environment, which can later on endanger life on this

planet. Given that the statistical testing for this particular dataset leaves room for speculation,

the experiment would either need to be repeated, or the parameters would need to be altered.
 Another property that was investigated to check the dependence on the amount of

nitrogen applied was the Soil Respiration levels over time. In order to analyze the dependence

more closely, a bar graph with the amount of carbon respired per unit area for each group of

hardwood and pine trees, along with their corresponding t-scores, were utilized. Soil respiration

is known to be a direct result of activities of soil organisms, in particular bacteria that are

characterized as the most abundant soil microorganisms (Gasparatos et al. 2011). Thus, the

correlation between nitrogen and soil respiration levels can be explained in terms of bacterial

richness and diversity associated with particular nitrogen additions (Yang et al., 2018). It can be

seen from the t-scores for control groups of both hardwood and pine trees that for the time

period from 1988 to 1989 there was a significant increase in the soil respiration rate, and even

though during the experiment no additional nitrogen was applied to that stands, such an effect

could be attributed to the long-lasting nitrogen natural cycle disruption caused by anthropogenic

factors. Moreover, for the same group of both types of trees, it cannot be claimed that there was

a notable difference in the soil respiration during the 1989-2001 time span since the t-scores

were either slightly above or below 1, indicating either “tension” or complete agreement. From

this fact, it can already be observed that there exists some kind of distinction between short-

and long-term effects of alternating nitrogen application. This distinction can be more clearly

observed in the low and high N groups, for which nitrogen was artificially added. The low

nitrogen groups for both trees were characterized with a considerable decline in the soil

respiration rate for the longer 1989-2001 time period, even though there was a discrepancy for

the first time span: a small increase for hardwood and no difference for pine trees. Even more

substantial long-term effect can be detected in the high nitrogen groups with relatively higher t-

scores that represent noticeable reduction in soil respiration rates. These results align with

those of the previous studies, which concluded that a significant decrease in the number of
bacterial microorganisms was a consequence of the long-term continuous input of nitrogen

fertilizers (Zhang et al., 2016). Nevertheless, there is still the opposite short-term effect that

requires explanation. Short-term nitrogen addition affected bacterial species through altering

NO3– components of the soil, which resulted in a moderate increase in the relative abundance of

various types of bacteria (Ning et al.). This can be assumed to be the primary cause of the

short-term soil respiration rate growth. However, in the long run, the reducing effect that N

fertilization has on the soil pH takes precedence, which results in the opposite effect of decline

in richness of bacterial microorganisms. Finally, considering these facts, it can be understood

why the artificial addition of nitrogen had a more substantial impact in the long run in low and

high N groups in comparison with that of the control group. Since the anthropogenic factors,

which were assumed to be the primary cause of the phenomenon, had been present for a more

considerable time comparing with the time span of the experiment, it can be deduced that

ecosystems, including bacterial microorganisms, that were a part of the control group had

already adapted to process increased levels of nitrogen.

Lastly, the effect of nitrogen concentration on microbial biomass helps determine the

impact of human activities on the environment. This parameter indicates the mass of living

components and organisms in the soil, which indirectly refers to the amount of bacteria and

fungi in the environment. This can be determined by the amount of carbon dioxide respired by

organisms in a gram of soil (the more they respire, the more biomass is produced). The

experiment consists of comparing samples with different levels of nitrogen in hardwood and pine

trees in a single year, to see if the amount of carbon differs in the short term. Results show no

connection between the fungal biomass and nitrogen levels in pine samples, and only a slight

correlation for fungal biomass in hardwood samples. However, the statistical results are not

strong enough to draw conclusions on such impact of human activities. In previous discussions,

it was shown that nitrogen addition has a positive effect on bacterial diversity on the short term
 but a negative effect on the long-term. Therefore, the experiment is not useful in determining a

clear relationship, but other research has proven that nitrogen addition has no long-term effects

on microbial biomass (Liu et al. 2015).

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