Tree hole odonates as environmental monitors: Non-invasive isolation of polymorphic microsatellites from the neotropical damselfly Megaloprepus caerulatus

摘要

Because of their complex mating behaviour and life cycle (alternating aquatic and terrestrial stages) odonates provide important model systems for environmental monitoring, evolutionary ecology, and conservation genetics. Many odonate species are endangered and call for the use of non-invasive molecular studies. In the neotropical damselfly Megaloprepus caerulatus we have identified polymorphic microsatellite loci by means of the randomly amplified microsatellite technique (RAMS; Ender et al. 1996). Using the DNA from each a single leg of three unrelated individuals we screened 63 RAPD primers for small size banding patterns. A total of 95 RAPD profiles was hybridized with digoxigenin labelled di- and trinucleotide repeats (GA(n), GT(n), CA(n) and AAT(n)) and 36 RAPD fragments harbouring microsatellite motifs were isolated. Cloning and sequencing of positive fragments revealed five polymorphic microsatellite loci. Since Megaloprepus caerulatus is a viable bio-indicator for primary rainforests the microsatellite system can be used to study the effects of forest fragmenation on population viability.