Search Results (3)

The molecular fingerprinting methods used to evaluate soil microbial diversity could also be used as effective biosensors for the purposes of monitoring ecological soil status. The biodiversity of microorganisms is a relevant index of soil activity and there is a necessity to develop tools to generate reliable results for an emerging approach in the field of environmental control using microbial diversity biosensors. This work reports a method under development for determining soil microbial diversity using high efficiency Multiplex PCR-Terminal Restriction Fragment Length Polymorphism (M-T-RFLP) for the simultaneous detection of bacteria, archaea and fungi. Three different primer sets were used in the reaction and the analytical conditions were optimized. Optimal analytical conditions were achieved using 0.5 µM of primer for bacteria and 1 µM for archaea and fungi, 4 ng of soil DNA template, and HaeIII restriction enzyme. Comparative tests using the proposed analytical approach and a single analysis of each microorganism group were carried out to indicate that both genetic profiles were similar. The Jaccard similarity coefficient between single and multiplexing approach ranged from 0.773 to 0.850 for bacteria and fungi, and 0.208 to 0.905 for archaea. In conclusion, the multiplexing and pooling approaches significantly reduced the costs and time required to perform the analyses, while maintaining a proper effectiveness. Full article

Soil microbial communities are involved in the maintenance of productivity and health of agricultural systems; therefore an adequate understanding of soil biodiversity plays a key role in ensuring sustainable use of soil. In the present study, we evaluated the influence of different cropping systems on the biodiversity of the soil bacterial communities, based on a 54-year field experiment established in Martonvásár, Hungary. Terminal restriction fragment length polymorphism (T-RFLP) fingerprinting technique was used to assess soil bacterial diversity and community structure in maize monoculture and three different crop rotations (maize–alfalfa, maize–wheat and the maize–barley–peas–wheat Norfolk type). No differences in richness and diversity were detected between maize monoculture and crop rotations except for the most intense rotation system (Norfolk-type). Although the principal component analysis did not reveal a clear separation between maize monoculture and the other rotation systems, the pairwise tests of analysis of similarity (ANOSIM) revealed that there are significant differences in the composition of bacterial communities between the maize monoculture and maize–alfalfa rotation as well as between wheat–maize and Norfolk-type rotation. Full article

This study examined seasonal differences in microbial community structure in the sediment of three streams in North Carolina’s Neuse River Basin. Microbes that reside in sediment are at the base of the food chain and have a profound influence on the health of freshwater stream environments. Terminal-Restriction Fragment Length Polymorphism (T-RFLP), molecular fingerprint analysis of 16S rRNA genes was used to examine the diversity of bacterial species in stream sediment. Sediment was sampled in both wet and dry seasons from an agricultural (Bear), mixed urban (Crabtree) and forested (Marks) Creek, and the microbiota examined. Gamma, Alpha and Beta proteobacteria were prevalent species of microbial taxa represented among all sites. Actinobacteria was the next most prevalent species observed, with greater occurrence in dry compared to the wet season. Discernable clustering was observed of Marks and Bear Creek samples collected during the wetter period (September–April), which corresponded with a period of higher precipitation and cooler surface water temperatures. Although not statistically significant, microbial community structure appeared different between season (ANOSIM, R = 0.60; p < 0.10). Principal components analysis confirmed this pattern and showed that the bacterial groups were separated by wet and dry seasonal periods. These results suggest seasonal differences among the microbial community structure in sediment of freshwater streams and that these communities may respond to changes in precipitation during wetter periods. Full article