TY  - THES
T1  - Effect of reduced nitrogen deposition on microbial activity, abundance and diversity in forest soils
A1  - Enowashu,Esther Eneckeh
Y1  - 2013/02/25
N2  - The deposition of nitrogen has increased many-fold due to anthropogenic activities. Since forest ecosystems are often limited by N availability, elevated N inputs from the atmosphere can have a fertilization effect but in the long-term, excess N can influence above- and below-ground production. One of the consequences of N deposition and increased N inputs is a shift in microbial community structure and function as ecosystems move towards N saturation. Soil microorganisms through the action of enzymes play an important role in N dynamics. Thus, the availability and turnover of N depends strongly on microbial abundance, diversity and activity which are in turn influenced by soil properties. Studies on the effects of high nitrogen inputs and the response of forest ecosystems to nitrogen saturation are many and well understood. However, the reversibility of N-induced shifts in forest ecosystem processes is largely unknown. This thesis was therefore designed to study the response of soil microorganisms to reduced N deposition. A biphasic approach was employed to look into (i) the general microbial functional status of the Solling forest site as well as (ii) the microbial community structure which may be a key regulator of two important processes of N transformation: denitrification and proteolysis.
The goal of the present thesis was addressed in three studies. Denitrification is considered sensitive to environmental changes and the response of nitrate-reducers and denitrifiers to reduced N deposition was determined in the first study. The goal of the second study was to investigate the overall microbial activity of the Solling forest profiles especially focussing on enzymes involved in the N cycle. This revealed a pronounced activity of peptidases whereby a set of novel pepN primers encoding alanine aminopeptidase enzyme was designed in the third study to determine the group of bacteria involved in proteolysis in forest as well as agricultural and grassland soils.
The Solling experimental station was established more than two decades ago and it gave the opportunity to study the N cycle in a natural forest ecosystem at different sampling dates and depths. A combination of classical biological methods and modern molecular techniques were used in the studies. Soil physico-chemical parameters (OC, Nt, NO3-, NH4+, pH, % Water content) were analysed to gain more information on mineralization and immobilization of N in the soil profiles. The analysis of microbial biomass, ergosterol content and the activity of several enzymes of the N, C and P cycles as well as enzyme activity of nitrate reducers was determined in order to interpret microbial functions. The abundance of nitrate reducers and denitrifiers were determined by quantitative PCR of 16S rRNA, nitrate reductase (narG and napA) and denitrification (nirK, nirS and nosZ) genes. The diversity of peptide degrading bacteria was analysed by PCR, cloning and sequencing and the construction of pepN gene libraries.  
The results of the first study indicated that time and space were the main drivers influencing the abundance and activity of the nitrate reducers and denitrifier communities in the forest soil profiles. Reduced N deposition had a of minimal effect. Interestingly, the ratios of nosZ to16S rRNA gene and nosZ to nirK increased with soil depth thereby tempting to conclude that the size of denitrifiers capable of reducing N2O into N2 might be bigger in the mineral horizons. In the second study, a stronger response of N cycling enzymes to reduced N deposition could be seen. However, these responses especially that of specific peptidases differed in magnitude which could be indicative of a modification of the reaction rates of the different N cycling enzymes. Correlation of nutrients (N, C, P) with microbial biomass and enzyme activities in the soil profiles revealed that substrate availability was the main factor influencing microbial activity. In the third study, analyses of gene libraries from extracted DNA from forest, agricultural and glacier soil samples revealed a high diversity of pepN sequences related to mainly α-Proteobacteria. A majority of the sequences showed similarity to published data revealing that the amplified region of pepN might be conserved. Linking diversity and enzymatic data, lowest diversity was observed in the agricultural soil where activity levels of alanine aminopeptidase were lowest indicating the importance of diversity studies for ecosystem functioning.
In conclusion, this thesis offers valuable contributions to understanding the impact of N deposition. The approach used was suitable to assess the response of the different microbial communities to reduced N deposition. The magnitude of the response depended strongly on space, time and substrate availability in soils as well as their interactions.  
KW  - Proteolyse 
KW  - Enzymaktivität
KW  - Mikrobielle Abundanz und Diversität
KW  - Denitrifikation
KW  - Waldböden
CY  - Hohenheim
PB  - Kommunikations-, Informations- und Medienzentrum der Universität Hohenheim
AD  - Garbenstr. 15, 70593 Stuttgart
UR  - http://opus.uni-hohenheim.de/volltexte/2013/811
ER  -