| Summary: | Premise of the study: New microsatellite (simple sequence repeat [SSR]) primers were developed from <i>Eucalyptus</i> expressed sequence tags (ESTs) and optimized for genetic studies of the southwestern Australian tree <i>E. gomphocephala</i>, which is severely impacted by tree health decline and habitat fragmentation.
Methods and Results: A total of 133 gene-homologous EST-SSR primer pairs were designed for <i>Eucalyptus</i>, and 44 were screened in <i>E. gomphocephala</i>. Of these, 17 produced reliable amplification products and 11 were polymorphic. Between two and 13 alleles were observed per locus, and observed heterozygosities ranged from 0.172 to 0.867. All 17 EST-SSRs that amplified <i>E. gomphocephala</i> cross-amplified to at least one of <i>E. marginata</i>, <i>E. camaldulensis</i>, and <i>E. victrix</i>.
Conclusions: This set of EST-SSR primer pairs will be valuable tools for future population genetic studies of <i>E. gomphocephala</i> and other eucalypts, particularly for studying gene-linked variation and informing seed-sourcing strategies for ecological restoration.
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