Fine-tuning for the tropics: application of eDNA technology for invasive fish detection in tropical freshwater ecosystems.

Invasive species pose a major threat to aquatic ecosystems. Their impact can be particularly severe in tropical regions, like those in northern Australia, where >20 invasive fish species are recorded. In temperate regions, environmental DNA (eDNA) technology is gaining momentum as a tool to detect aquatic pests, but the technology's effectiveness has not been fully explored in tropical systems with their unique climatic challenges (i.e. high turbidity, temperatures and ultraviolet light). In this study, we modified conventional eDNA protocols for use in tropical environments using the invasive fish, Mozambique tilapia (Oreochromis mossambicus) as a detection model. We evaluated the effects of high water temperatures and fish density on the detection of tilapia eDNA, using filters with larger pores to facilitate filtration. Large-pore filters (20 µm) were effective in filtering turbid waters and retaining sufficient eDNA, whilst achieving filtration times of 2-3 min per 2-L sample. High water temperatures, often experienced in the tropics (23, 29, 35 °C), did not affect eDNA degradation rates, although high temperatures (35 °C) did significantly increase fish eDNA shedding rates. We established a minimum detection limit for tilapia (1 fish/0.4 megalitres/after 4 days) and found that low water flow (3.17 L/s) into ponds with high fish density (>16 fish/0.4 megalitres) did not affect eDNA detection. These results demonstrate that eDNA technology can be effectively used in tropical ecosystems to detect invasive fish species.

Microsatellite markers in the Australian desert plant, Solanum centrale (Solanaceae).

PREMISE OF THE STUDY: The first microsatellite primers were developed for Solanum centrale, a native Australian species used in the bush foods industry. Existing markers developed for other Solanum species were also amplified. METHODS AND RESULTS: Using an enrichment cloning protocol, seven novel markers were developed, and 48 existing markers from other Solanum species were tested, resulting in the characterization of a set of six highly polymorphic co-dominant loci for use in S. centrale. Microsatellite screening revealed polyploidy. Among the six highly polymorphic loci, allelic diversity ranged from 7 to 14. CONCLUSIONS: These markers will be useful for investigating genetic diversity and as a simple way of estimating ploidy of wild populations.

Isolation and characterization of microsatellite loci in Santalum lanceolatum and Santalum leptocladum (Santalaceae).

PREMISE OF THE STUDY: Microsatellite primers were developed for the first time in the native Australian sandalwood species Santalum lanceolatum. • METHODS AND RESULTS: Using an enrichment cloning protocol, five novel polymorphic codominant loci were developed and characterized in S. lanceolatum and S. leptocladum. In addition to these, three existing microsatellite loci from other sandalwood species were successfully amplified and characterized for S. lanceolatum and S. leptocladum. Among the eight loci, allelic diversity ranged from 4 to 29. • CONCLUSIONS: Primers will be useful for studies of clonality, genetic diversity and spatial genetic structure in wild populations. When coupled with other molecular techniques will help investigate the relationship between S. lanceolatum and S. leptocladum, species of commercial and conservation interest.