Detection and Quantification of Nitrifying Bacteria Using Real-Time PCR.

Nitrification is the microbial-mediated transformation of ammonium (NH4+) into nitrate (NO3-). Many plant species depend on the availability of NO3- as the main source of nitrogen (N). On the other hand, because NO3- is highly mobile in the soil profile, its excess concentration can cause environmental pollution. Nitrification can be estimated at the process level, but with the development of molecular techniques it is also possible to estimate the abundance of nitrifying bacteria in the soil. Hence, in this chapter we describe the procedure for detection and quantification of nitrifying bacteria in soil samples using real-time quantitative polymerase chain reaction (PCR).

Iron-mediated stabilization of soil carbon amplifies the benefits of ecological restoration in degraded lands.

Recent observations across a 14-year restoration chronosequence have shown an unexpected accumulation of soil organic carbon in strip-mined areas of central Brazil. This was attributed to the rapid plant colonization that followed the incorporation of biosolids into exposed regoliths, but the specific mechanisms involved in the stabilization of carbon inputs from the vegetation remained unclear. Using isotopic and elemental analyses, we tested the hypothesis that plant-derived carbon accumulation was triggered by the formation of iron-coordinated complexes, stabilized into physically protected (occluded) soil fractions. Confirming this hypothesis, we identified a fast formation of microaggregates shortly after the application of iron-rich biosolids, which was characterized by a strong association between pyrophosphate-extractable iron and plant-derived organic matter. The formation of microaggregates preceded the development of macroaggregates, which drastically increased soil carbon content (-140 Mg C/ha) a few years after restoration. Consistent with previous theoretical work, iron-coordinated organic complexes served as nuclei for aggregate formation, reflecting the synergistic effect of biological, chemical, and physical mechanisms of carbon stabilization in developing soils. Nevertheless, iron was not the only factor affecting soil carbon content. The highest carbon accumulation was observed during the period of highest plant diversity (> 30 species; years 3-6), declining significantly with the exclusion of native species by invasive grasses (years 9-14). Furthermore, the increasing dominance of invasive grasses was associated with a steady decline in the concentration of soil nitrogen and phosphorus per unit of accumulated carbon. These results demonstrate the importance of interdependent ecological and biogeochemical processes, and the role of soil-plant interactions in determining the success of restoration efforts. In contrast with previous but unsuccessful attempts to restore mined areas through nutrient application alone, iron-mediated stabilization of vegetation inputs favored the regeneration of a barren stable state that had persisted for over five decades since disturbance. The effectiveness of coupled organic matter and iron "fertilization," combined with management of invasive species, has the possibility to enhance terrestrial carbon sequestration and accelerate the restoration of degraded lands, while addressing important challenges associated with urban waste disposal.

Unprecedented carbon accumulation in mined soils: the synergistic effect of resource input and plant species invasion.

Opencast mining causes severe impacts on natural environments, often resulting in permanent damage to soils and vegetation. In the present study we use a 14-year restoration chronosequence to investigate how resource input and spontaneous plant colonization promote the revegetation and reconstruction of mined soils in central Brazil. Using a multi-proxy approach, combining vegetation surveys with the analysis of plant and soil isotopic abundances (delta13C and delta15N) and chemical and physical fractionation of organic matter in soil profiles, we show that: (1) after several decades without vegetation cover, the input of nutrient-rich biosolids into exposed regoliths prompted the establishment of a diverse plant community (> 30 species); (2) the synergistic effect of resource input and plant colonization yielded unprecedented increases in soil carbon, accumulating as chemically stable compounds in occluded physical fractions and reaching much higher levels than observed in undisturbed ecosystems; and (3) invasive grasses progressively excluded native species, limiting nutrient availability, but contributing more than 65% of the total accumulated soil organic carbon. These results show that soil-plant feedbacks regulate the amount of available resources, determining successional trajectories and alternative stable equilibria in degraded areas undergoing restoration. External inputs promote plant colonization, soil formation, and carbon sequestration, at the cost of excluding native species. The introduction of native woody species would suppress invasive grasses and increase nutrient availability, bringing the system closer to its original state. However, it is difficult to predict whether soil carbon levels could be maintained without the exotic grass cover. We discuss theoretical and practical implications of these findings, describing how the combination of resource manipulation and management of invasive species could be used to optimize restoration strategies, counteracting soil degradation while maintaining species diversity.