Newcastle University ePrints -eprint.ncl.ac.uk Logan SA, Phuekvilai P, Wolff K. Ancient woodlands in the limelight: delineation and genetic structure of ancient woodland species Tilia cordata and Tilia platyphyllos (Tiliaceae) in the UK. Abstract Ancient woodlands in Europe are currently subjected to fragmentation, which leads to ecological deterioration and genetic isolation. This is likely to intensify with further climate change and increased land use. Tilia are keystone, ancient woodland indicators, represented in the United Kingdom by two species (Tilia cordata and T. platyphyllos) and their hybrid (T. x europaea). For management of the species we need to investigate genetic differentiation and population genetic structure. Morphology can usually separate the two species but the distinguishing features are not always observable, particularly in less optimum, shady, conditions. Here we test for the first time, 13 microsatellite markers to discriminate the species and hybrid, and to assess the population genetic diversity of the two species. The markers successfully distinguished the species and hybrid. The two species show significant genetic differentiation, indicating two distinct biological units. Sub-structure within both species is loosely related to geographic location. Because Tilia populations in the UK are highly fragmented future conservation should consider provenance issues.

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• Premise of the study: Microsatellite markers in the genus Tilia were developed to investigate the genetic variation in T. platyphyllos and its relationship with T. cordata.• Methods and Results: Fifteen microsatellite markers were developed using a microsatellite enrichment protocol. Most loci show a high level of polymorphism in two T. platyphyllos populations from France. The number of alleles ranged from one to 15, with a mean of 8.96. The mean observed and expected heterozygosities were 0.71 and 0.70, respectively. Cross-amplification results indicated that 12 out of 15 loci amplified polymorphic loci in 23 species in the genus.• Conclusions: These markers will be useful tools for investigating the phylogeography and hybridization of Tilia species.

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