Vertical distribution of ammonia-oxidizing microorganisms across a soil profile of the Chinese Loess Plateau and their responses to nitrogen inputs.

Ammonia-oxidizing archaea (AOA) and bacteria (AOB) oxidize ammonia into nitrite, the first and rate-limiting step of microbial nitrification, and exert major controls over soil nitrogen transformations. The Loess Plateau in northwest China is characterized with deep soils that are often exposed to the surface and reactive nitrogen (N) inputs due to erosion and human removal of the surface soil. However, few have examined the distribution of AOA and AOB along the profile of Loess Plateau soils and their responses to N inputs. We examined the abundance and diversity of AOA and AOB along the soil profile (0-100cm) and their responses to two levels of N inputs (low at 10, and high at 100µgNg-1 soil) in a 55-d incubation experiment. While AOB were most numerous in the surface soil (0-20cm), AOA were most abundant in the subsoils (20-40 and 40-60cm), suggesting a niche differentiation between AOA and AOB along the soil profile. High N input increased AOB nearly ten-fold in the upper two layers of soils (0-20 and 20-40cm) and sixteen to twenty-five fold in the deeper soil layers (40-60, 60-80 and 80-100cm). However, it only increased AOA by 7% (40-60cm) to 48% (20-40cm). In addition, potential nitrification rate and N2O emissions correlated only with AOB. Finally, high N input significantly increased AOB diversity and led to nitrite accumulation in deep soil layers (60-80 and 80-100cm). Together, our results showed that high N input can significantly alter the diversity and function of ammonia-oxidizing microbes in the deep soil of Loess Plateau, suggesting the need to examine the generality of the observed changes and their potential environmental impacts.

In Situ Dark Adaptation Enhances the Efficiency of DNA Extraction from Mature Pin Oak (Quercus palustris) Leaves, Facilitating the Identification of Partial Sequences of the 18S rRNA and Isoprene Synthase (IspS) Genes.

Mature oak (Quercus spp.) leaves, although abundantly available during the plants' developmental cycle, are rarely exploited as viable sources of genomic DNA. These leaves are rich in metabolites difficult to remove during standard DNA purification, interfering with downstream molecular genetics applications. The current work assessed whether in situ dark adaptation, to deplete sugar reserves and inhibit secondary metabolite synthesis could compensate for the difficulties encountered when isolating DNA from mature leaves rich in secondary metabolites. We optimized a rapid, commercial kit based method to extract genomic DNA from dark- and light-adapted leaves. We demonstrated that in situ dark adaptation increases the yield and quality of genomic DNA obtained from mature oak leaves, yielding templates of sufficiently high quality for direct downstream applications, such as PCR amplification and gene identification. The quality of templates isolated from dark-adapted pin oak leaves particularly improved the amplification of larger fragments in our experiments. From DNA extracts prepared with our optimized method, we identified for the first time partial segments of the genes encoding 18S rRNA and isoprene synthase (IspS) from pin oak (Quercus palustris), whose full genome has not yet been sequenced.