The dynamics of indigenous bacterial and fungal soil communities were followed throughout the decomposition of wheat straw residue. More precisely, such dynamics were investigated in the different soil zones under the influence of decomposing wheat straw residue (i.e. residues, soil adjacent to residue = detritusphere, and bulk soil). The genetic structures of bacterial and fungal communities were compared throughout the decomposition process long by applying B- and F-ARISA (for bacterial and fungal-automated ribosomal intergenic spacer analysis) to DNA extracts from these different zones. Residue decomposition induced significant changes in bacterial and fungal community dynamics with a magnitude of changes between the different soil zones ordered as followed: residue > detritusphere > bulk soil, confirming the spatial structuration of the sphere of residue influence to the 4–6 mm soil zone in contact with residue. Furthermore, significant differences in the structure of bacterial and fungal communities were apparent between the early (14 and 28 days) and late (from 56 to 168 days) stages of decomposition. These could be related to ecological attributes such as the succession of r- (copiotrophs) and K- (oligotrophs) strategists. Microbial diversity at the early (28 days) and late (168 days) stages of degradation was further analysed by a molecular inventory of 16S and 18S rDNA in DNA extracts from the residue zone. This confirmed the succession of different populations during residue decomposition. Fluorescent Pseudomonas spp. and Neurospora sp. were dominant in the early stage with subsequent stimulation of Actinobacteria and Deltaproteobacteria taxa, as well as Basidiomycota fungal taxa and Madurella spp. According to the ecological attributes of these populations, microbial succession on fresh organic residue incorporated in soil would be dominated by copiotrophs and r-strategists in the early stages, with oligotrophs (K-strategists) increasing in relative abundance as substrate quantity and/or quality declines over time.

Impact of wheat straw decomposition on successional patterns of soil microbial community structure

Fungi constitute an important group in soil biological diversity and functioning. However, characterization and knowledge of fungal communities is hampered because few primer sets are available to quantify fungal abundance by real-time quantitative PCR (real-time Q-PCR). The aim in this study was to quantify fungal abundance in soils by incorporating, into a real-time Q-PCR using the SYBRGreen (R) method, a primer set already used to study the genetic structure of soil fungal communities. To satisfy the real-time Q-PCR requirements to enhance the accuracy and reproducibility of the detection technique, this study focused on the 18S rRNA gene conserved regions. These regions are little affected by length polymorphism and may provide sufficiently small targets, a crucial criterion for enhancing accuracy and reproducibility of the detection technique. An in silico analysis of 33 primer sets targeting the 18S rRNA gene was performed to select the primer set with the best potential for real-time Q-PCR: short amplicon length; good fungal specificity and coverage. The best consensus between specificity, coverage and amplicon length among the 33 sets tested was the primer set FR1/FF390. This in silico analysis of the specificity of FR1/FF390 also provided additional information to the previously published analysis on this primer set. The specificity of the primer set FR1/FF390 for Fungi was validated in vitro by cloning - sequencing the amplicons obtained from a real time Q-PCR assay performed on five independent soil samples. This assay was also used to evaluate the sensitivity and reproducibility of the method. Finally, fungal abundance in samples from 24 soils with contrasting physico-chemical and environmental characteristics was examined and ranked to determine the importance of soil texture, organic carbon content, C:N ratio and land use in determining fungal abundance in soils.

/pdf/validation-and-application-of-a-pcr-primer-set-to-quantify-3hy3het8au.pdf

Validation and Application of a PCR Primer Set to Quantify Fungal Communities in the Soil Environment by Real-Time Quantitative PCR

Bacteria in Agrobiology: Plant Growth Responses

Background: Copper mining has led to Cu pollution in agricultural soils. In this report, the effects of Cu pollution on bacterial communities of agricultural soils from Valparaiso region, central Chile, were studied. Denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA genes was used for the characterization of bacterial communities from Cu-polluted and non-polluted soils. Cu-resistant bacterial strains were isolated from Cu-polluted soils and characterized. Results: DGGE showed a similar high number of bands and banding pattern of the bacterial communities from Cu-polluted and non-polluted soils. The presence of copA genes encoding the multi-copper oxidase that confers Cu-resistance in bacteria was detected by PCR in metagenomic DNA from the three Cu-polluted soils, but not in the non-polluted soil. The number of Cu-tolerant heterotrophic cultivable bacteria was significantly higher in Cu-polluted soils than in the non-polluted soil. Ninety two Cu-resistant bacterial strains were isolated from three Cu-polluted agricultural soils. Five isolated strains showed high resistance to copper (MIC ranged from 3.1 to 4.7 mM) and also resistance to other heavy metals. 16S rRNA gene sequence analyses indicate that these isolates belong to the genera Sphingomonas, Stenotrophomonas and Arthrobacter. The Sphingomonas sp. strains O12, A32 and A55 and Stenotrophomonas sp. C21 possess plasmids containing the Cu-resistance copA genes. Arthrobacter sp. O4 possesses the copA gene, but plasmids were not detected in this strain. The amino acid sequences of CopA from Sphingomonas isolates (O12, A32 and A55), Stenotrophomonas strain (C21) and Arthrobacter strain (O4) are closely related to CopA from Sphingomonas, Stenotrophomonas and Arthrobacter strains, respectively. Conclusions: This study suggests that bacterial communities of agricultural soils from central Chile exposed to long-term Cu-pollution have been adapted by acquiring Cu genetic determinants. Five bacterial isolates showed high copper resistance and additional resistance to other heavy metals. Detection of copA gene in plasmids of four Cu-resistant isolates indicates that mobile genetic elements are involved in the spreading of Cu genetic determinants in polluted environments.

https://link.springer.com/content/pdf/10.1186%2F1471-2180-12-193.pdf

Characterization of copper-resistant bacteria and bacterial communities from copper-polluted agricultural soils of central Chile

Copper removal and microbial community analysis in single-chamber microbial fuel cell.

Fingerprinting and diversity of bacterial copA genes in response to soil types, soil organic status and copper contamination.

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Fingerprinting and diversity of bacterial copA genes in response to soil types, soil organic status and copper contamination.