Variability of nitrogen-cycle microbial communities determined by the age of restored wetlands
Authors: Kasak Kuno , Anthony Tyler , Valach Alex , Hemes Kyle , Kill Keit , Silver Whendee , Mander Ülo , Szutu Daphne , Verfaillie Joseph , Baldocchi Dennis 
Published: March 2020

&lt;p&gt;&lt;span&gt;Restoring degraded peat soils to wetlands can be an attractive and efficient measure with many benefits including carbon sequestration, water quality improvement, food and habitat for wildlife, flood control, and opportunities for recreation. Agricultural lands which are restored to wetlands will start rebuild soils and reverse land subsidence. Using eddy covariance towers in four wetlands that were restored in 1997, 2010, 2013 and 2016 in the Sacramento-San Joaquin Delta in California, we saw high carbon sequestration potentials and peat accumulation. Since soil restoration takes place gradually, it is important to specify the critical turning-points in the process of improving soil microbial community structure and nitrogen cycling. In August 2018, soil samples from four wetlands with different restoration ages in the Delta were collected for chemical and microbial analyses. The bacterial and archaeal 16S rRNA genes and functional genes involved in nitrogen cycling (&lt;em&gt;nirS&lt;/em&gt;, &lt;em&gt;nirK&lt;/em&gt;, &lt;em&gt;nosZ-I&lt;/em&gt;, &lt;em&gt;nosZ-II&lt;/em&gt;, bacterial and archaeal &lt;em&gt;amoA&lt;/em&gt;, &lt;em&gt;nifH&lt;/em&gt;, &lt;em&gt;nrfA&lt;/em&gt;, and ANAMMOX-specific genes) in soils were determined using a quantitative PCR method. Soil chemical parameters such as C%, N%, Al, Mn, Fe and two different organic and inorganic P pools were analysed as well. Preliminary results indicate significant dissimilarities in the abundance of soil bacterial and archaeal communities, as well as &lt;em&gt;nirS&lt;/em&gt;, &lt;em&gt;nirK&lt;/em&gt;, &lt;em&gt;nosZ&lt;/em&gt;, &lt;em&gt;nifH&lt;/em&gt;, &lt;em&gt;nrfA&lt;/em&gt; and archaeal &lt;em&gt;amoA&lt;/em&gt; gene-possessing microbial communities in different wetlands. Data analysis showed several statistically significant relationships between target gene parameters and soil chemical parameters that were different when comparing the sites with the restoration age. It is clear, that the complexity of the relationships increases as the wetland gets older. For example, in younger wetlands the availability of C and N plays a crucial role in gene abundances while in the oldest wetland, the most important chemical parameters were different phosphorus forms. This might indicate that more than 20 years of C and N accumulation has led to the availability of phosphorus for N transformation now to be the main limiting factor. Another important finding was that the design criteria can also determine how the wetland acts in terms of nitrogen gas emissions. For example, one of the wetlands was designed with more varied bathymetry that includes many open channels and a fluctuating water table. We saw that the &lt;em&gt;nifH&lt;/em&gt; gene-possessing microbes that are responsible for molecular N fixing are highly abundant in open water areas while at the same time this wetland has also the highest abundance of &lt;em&gt;nir&lt;/em&gt; genes that control N&lt;sub&gt;2&lt;/sub&gt;O production by denitrifiers. Our study demonstrates that the design of the wetland can have a significant impact on N-transforming processes, but most importantly at some age, restored wetlands become more similar to natural wetlands.&lt;/span&gt;&lt;/p&gt;