Climate warming is predicted to raise the mean global temperature by 1 °C in the next 50 years, and this change is believed to be capable of affecting soil organic matter cycling and nutrient availability. With the aim of increasing knowledge on the response of forest soils to the ongoing climate change, we used altitude as a proxy for temperature change and studied chemical and biochemical properties of European beech (Fagus sylvatica L.) forest soils at two altitudes (800 and 1000 m) from central Apennines (Italy). Results showed that 1 °C of mean annual air temperature difference between the sites at the two altitudes had greater effect on the mineral horizons than on the organic horizons. At higher altitude, mineral soil had limited development, higher pH, and higher organic matter content due to the lower efficiency of the microbial community. Enzymatic activities of the organic horizons were generally not affected by altitude. Conversely, we observed a higher activity of xylosidase, β-glucosidase, alkaline phosphomonoesterase, arylsulfatase, and leucine-aminopeptidase in the sub-superficial horizons (Bw1 and Bw2) of the soils at 1000 m. We hypothesized that, as a response to environmental and climatic constraints occurring at higher altitude, plant roots increase the production of enzymes directly and/or indirectly by triggering the microbial community through exudation.

Characterisation and evaluation of soil bacteria were conducted in order to select the most potent strains that participate in the degradation of cellulose in unique agroecosystem and climatic conditions. Cellulolytic activity of soil bacteria was estimated using qualitative assays such as growth on selective media followed by screening with Congo red, Gram’s iodine solution, confirmation test on Congo red agar, determination of enzyme production, and sugar utilisation pattern. A total of 159 soil cellulolytic bacterial strains were selected based on shape, size, and colony characteristics. According to the results of all three screening assays, sixty-four, thirty-eight and fifty-one isolates were able to degrade at some level of cellulose, respectively. Partial sequencing of the 16S rRNA gene of 64 bacterial strains obtained using sequences retrieved from the databases indicated the presence of cellulolytic bacteria represented by members of the phyla Actinobacteria (48.44%), followed by Firmicutes (32.81%), Proteobacteria (15.62%) and Bacteroidetes (3.13%). Determination of enzyme production showed that fifteen strains possess endoglucanases activity which ranged from 9.09 to 942.41 nanomoles of MUF (4-methylumbelliferone) mL. Likewise, β-glucosidase enzyme activity was determined in 23.4 % of all isolates. The sugar utilisation pattern of soil bacterial strains displayed the different capabilities of growth and utilisation on various carbon sources, which occur in lignocellulosic materials (cellulose, starch) or their hydrolysates (glucose, galactose, fructose, cellobiose, maltose, lactose, sucrose, trehalose).

DNA-based technologies have become widespread tools for soil microbiological analyses in recent years. DNA extraction from the soil is a key step for these approaches: it is a challenge for researchers as it is still both expensive and time-consuming when large surveys are planned. The aim of this study was to develop a high-throughput automated protocol for DNA extraction and purification from soil. The protocol was based on the BioSprint 96 platform and compared for validation with another automated procedure and two commercial column-based kits. To evaluate the performances of the protocols, we considered quality, quantity, and amplifiability of the isolated DNA. The material isolated by means of the four protocols showed appropriate yield and quality and positive amplification. The isolation protocol presented here provided similar results to those of the commercial kits but with two essential differences: cost and time for DNA extraction were drastically reduced. This rapid and efficient protocol is envisaged as ideal to standardize soil studies and treat large numbers of samples, representing a workable alternative to low-throughput and expensive manual extraction methods.