Izolacija eksogene glivne DNA iz neinvazivnih vzorcev (iztrebkov) prostoživečih parkljarjev = Isolation of exogenous fungal DNA from non-invasive samples (faeces) of wild ungulates
Duniš, Luka ...
Arbuscular mycorrhizal fungi are symbionts of a large majority of terrestrial plants. Along with other benefits of mycorrhiza, the symbiotic relationship allows exchange of nutrients among symbiotic ...partners. Large mammals with their metabolic processes (i.e., nutrition and excretion, zoohory) are important vectors for the transfer and dispersal of fungi within and between different environments. Their faeces are an important source of their DNA, as well as DNA from the species they eat and microbial DNA. However, isolating DNA from faeces is difficult and often unsuccessful and poses a major challenge for laboratory work. We tested the efficiency of two different DNA isolation kits (Qiagen QIAamp Fast DNA Stool Mini Kit, and Qiagen DNeasy PowerSoil Kit) for fungal DNA extraction from faeces of European roe deer (Capreolus capreolus) and red deer (Cervus elaphus). We estimated the efficiency of the kits by measuring the concentration of isolated DNA and by amplification success using the arbuscular mycorrhizal fungal specific NS31 and AM1 primers for the 18S region of ribosomal RNA. Fragments of 550 bp arbuscular mycorrhizal DNA were isolated and amplified, and we optimized the annealing temperatures using qPCR. In terms of better PCR amplification, our results showed that the Qiagen DNeasy PowerSoil Kit performed better than the Qiagen QIAamp Fast DNA Stool Mini Kit. By analysing the lengths of DNA fragments, we confirmed that wild ungulate faeces can be a useful tool in environmental genetics, for example as a source of fungal DNA, which allows us to study the potential animal dispersal of mycorrhizal fungi and other ecological interactions in terrestrial ecosystems (e.g. dietary preferences and consumption of fungi, reasons for wild boar rooting, transmission of fungi by zoohory).
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