Poster abstracts

Establishing DNA-Barcoding Methods in Diatoms for Diversity Assessments

B. Gemeinholzer, N. Enke & R. Jahn
Botanischer Garten und Botanisches Museum Berlin-Dahlem, Freie Universität Berlin, Germany

Microscopic algae are important bio-indicators to monitor water quality as they are sensitive to pollution and contamination, trophies, as well as acidification, and salinisation. Diatoms are especially suited for water quality assessments as it is the only group of microscopic algae which is present in all water sources.
Continuous screening of algal biodiversity can provide information about threatened ecosystem. Thus diatoms are an ideal model group to establish DNA-Barcoding methods to provide an easy to use, quick, efficient, standardised organism identification tool to serve routine water quality assessments, as morphological identification is time consuming and demands specialised in-depth knowledge.
Method optimisation is the main task of this project. A pilot study was carried out by identifying diatoms by light microscope from a water sample from the Tegeler See in Berlin/Germany. Total genomic DNA was isolated from the same sample, the 18S region was amplified using diatom specific priming sites and the fragments were cloned, picked, and sequenced. The sequences were checked against a molecular reference database (AlgaTerra). 62 different diatom species could be identified by means of light microscopy; in the same sample 59 species were detected by DNA-Barcoding methods. However, results were not completely congruent in both methods; but most taxa identified by molecular means were known to occur potentially in the sample site. This elucidates the potential of DNA-Barcoding for diversity and water quality assessments; but as well shows the need for standardised and optimised laboratory protocols. In subsequent trials, primer screenings were carried out to find a more suitable region for DNA Barcoding in diatoms, different (nondestructive) DNA extraction methods were applied and up to now, nothing is known about the number of clones required to completely cover the diatom diversity in fresh water samples.

The Use of DNA Barcoding in Forensic Entomology in Poland

Tadeusz Malewski, Agnieszka Draber-Monko, Wieslaw Bogdanowicz
Museum and Institute of Zoology, Polish Academy of Sciences, Warsaw, Poland

A carrion-fly maggot is the most common type of insect evidence collected during investigation of dead body. Determining the species composition of such sample is an important first step in a forensic entomological analysis. Although species-diagnostic anatomical characters have been reported for the eggs and early larvae of some taxa, an anatomical approach is rather difficult to apply for immature stages of several taxa. Mitochondrial DNA can be used in species identification of all life stages of a carrion fly. Recent study of the COI gene has identified as many as 45 regions that distinguish species of the three "forensic" genera of blowflies (family Calliphoridae) - Lucilla, Calliphora and Chrysomya. In Poland there are 66 species of blowflies of which twelve are of forensic value. In our institute we collected specimens and prepared reference DNA samples of all forensically important Calliphoridae. Most of these have been sequenced, and this information is available for use by police and other investigative agencies. Based on results we obtained  it will be possible to design the sequence-specific probes. Moreover, beyond the applications to forensic entomology, an understanding of the COI sequences of Calliphoridae may be also treated as a valuable addition to the DNA barcoding initiative.

Turning Sequences into Taxa

Jenna Mann, Mark Blaxter
Institute of Evolutionary Biology, Edinburgh University

Most animal taxa can be classified as meiofauna (body length less than 1mm). The number of meiofaunal ‘species’ is unknown, but is likely to be in the millions. In the absence of large communities of Linnaean taxonomists dedicated to meiofauna, how can these organisms be identified, taxonomised and analysed? We can acquire DNA barcode data from individual meiofaunal specimens. In most instances we do not have exemplar specimens identified to species, nor do we expect to be able to get sequence from type specimens. So how can we identify taxa independent of external reference samples? We generated a congruent barcode dataset for cytochrome oxidase 1 (CO1), large and small ribosomal subunits (LSU and SSU respectively) from a set of unidentified rhabditid nematodes. Here we investigate the use of different genes and methods in defining molecular operational taxonomic units (MOTU) in this nematode fauna..
 

Plant and Fungi Barcoding in Poland

Zbigniew Mirek, Wojciech Bieniek, Agnieszka Sztorc
W. Szafer Institute of Botany, Polish Academy of Sciences, Krakow

Merits and organizational issues related to The Polish Center of Plant and Fungi Barcoding in the Institute of Botany, Polish Academy of Sciences in Krakow are presented. Current stage of art and the action plan for coming years are characterized. Hopes and still existing open questions concerning plant and fungi barcoding in Poland are discussed.