Environmental changes in biodiversity hotspot ecosystems of
South Ecuador: RESPonse and feedback effECTs (FOR2730)

Resumen:

- Presently, molecular ecological studies of arbuscular mycorrhizal fungi (AMF) are only possible above species-level when targeting entire communities. To improve molecular species characterisation and to allow species-level community analyses in the field, a set of newly designed AMF specific PCR primers is successfully tested. - Nuclear rDNA fragments from diverse phylogenetic AMF lineages were sequenced and analysed to design four primer mixtures, each targeting one binding site in the small (SSU) or large subunit rDNA (LSU). To allow species resolution, they span a fragment covering the partial SSU, whole internal transcribed spacer rDNA region (ITS), and partial LSU. - The new primers are suitable to i) specifically amplify AMF rDNA from material that may be contaminated by other organisms, e.g., samples from pot cultures or the field, ii) characterise the diversity of AMF species from field samples, and iii) amplify a SSU-ITS-LSU fragment that allows phylogenetic analyses with species-level resolution. - The described PCR primers can be used for monitoring of entire AMF field communities, based on a single rDNA marker region. Their application will improve the base for deep sequencing approaches, and moreover they can be efficiently used as DNA barcoding primers.