Characterisation of polymorphic microsatellite loci in the freshwater bryozoan Fredericella sultana
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TECHNICAL NOTE
Characterisation of polymorphic microsatellite loci in the freshwater bryozoan Fredericella sultana H. Hartikainen • J. Jokela
Received: 11 November 2011 / Accepted: 18 November 2011 / Published online: 29 November 2011 Ó Springer Science+Business Media B.V. 2011
Abstract Eight polymorphic microsatellite loci were isolated from massively parallel next-generation sequencing data and tested in three populations (74 individuals) of the colonial freshwater bryozoan Fredericella sultana. Up to 13 alleles per locus were found and all loci were polymorphic in all populations. Minimum of three loci were sufficient to distinguish all unique multilocus genotypes. These highly variable markers are suitable for clonal identity assignment based on unique multilocus genotypes and provide tools for resolving fine scale population structure in a species characterised by clonal, vegetative growth and asexual reproduction. Keywords Bryozoa Myxozoa Proliferative kidney disease 454 Microsatellite Polymorphic
Freshwater bryozoans (Family Phylactolaemata, Phylum Bryozoa) are colonial suspension feeders with important ecological roles in nutrient cycling and structuring of benthic freshwater habitats (Sørensen et al. 1986). The bryozoan Fredericella sultana also acts as the primary host H. Hartikainen J. Jokela Department of Aquatic Ecology, EAWAG, Swiss Federal Institute of Aquatic Science and Technology, ¨ berlandstrasse 133, 8600 Du¨bendorf, Switzerland U H. Hartikainen J. Jokela ETH-Zu¨rich, Institute of Integrative Biology, Universita¨tstrasse 16, 8092 Zu¨rich, Switzerland Present Address: H. Hartikainen (&) Department of Zoology, Natural History Museum, Cromwell Road, London SW7 5BD, UK e-mail: [email protected]
for the myxozoan parasite Tetracapsuloides bryosalmonae, which causes the devastating proliferative kidney disease (PKD) of salmonid fish (Anderson et al. 1999). PKD affects many endangered salmon and trout species (Hedrick et al. 1993) and F. sultana has a key role in the persistence and spread of the PKD parasite (Okamura et al. 2011). Thus, understanding the population dynamics of F. sultana is crucial for explaining the recent emergence of PKD and may contribute to salmonid conservation and management. Fredericella sultana colonies grow and spread by budding new colony modules and can reach high densities in suitable conditions. Colonies produce asexual resting stages and may disperse via colony fragments. In contrast, sexual reproduction is short in duration and may be occasionally forgone (Wood 1973). Assignment of clonal identities within such potentially homogeneous populations can be conducted using unique multilocus genotypes (MLGs), but requires highly variable markers for confident assignment. We used newly developed (e.g. Castoe et al. 2010), rapid methods utilising massively parallel 454 sequencing technology to characterize hypervariable microsatellite loci in F. sultana for clone identification purposes. About 3 lg of genomic DNA was isolated from whole colonies using a
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