Molecular Ecology Notes (2005) 5, 542–545 doi: 10.1111/j.1471-8286.2005.00982.x © 2005 Blackwell Publishing Ltd Blackwell Publishing, Ltd. PRIMER NOTE Development and characterization of 11 microsatellite markers in a widespread Neotropical seasonally dry forest tree species, Geoffroea spinosa Jacq. (Leguminosae) Y. NACIRI-GRAVEN,* S. CAETANO,* D. PRADO,† R. T. PENNINGTON‡ and R. SPICHIGER* * Laboratoire de Génétique et Phylogénie Moléculaires, Conservatoire et Jardin Botaniques, Chemin de l’Impératrice 1, 1292 Chambésy Genève, Switzerland, † Cátedra de Botánica Morfológica y Sistemática, Facultad de Ciencias Agrarias, UNR, Casilla de Correo No. 14, S2125ZAA, Zavalla, Argentina, ‡ Tropical Diversity Section, Royal Botanic Garden Edinburgh, 20a Inverleith Row, EH3 5LR, Edinburgh, United Kingdom Abstract Eleven dinucleotide microsatellites were developed in Geoffroea spinosa (Leguminosae), a widespread tree of the seasonally dry Neotropical forests, and characterized on six popula- tions from Peru, Argentina and Paraguay. Four of them amplified on the Peruvian popula- tions only, probably because of mutations in the microsatellite flanking regions in the other populations. Ten microsatellites were found polymorphic, with within population gene diversities ranging from 0.17 to 0.95, and a number of alleles varying from seven to 19. A significant overall genetic differentiation was also found (θ = 0.212; P < 0.01). Keywords: Geoffroea spinosa, microsatellite, population genetics, seasonally dry tropical forest Received 17 December 2004; revision received 2 February 2005; accepted 25 February 2005 Seasonally dry forests may be defined as woody veget- ation occurring in areas with a dry season lasting up to half a year in one or two periods, and an annual rainfall of between 400 and 1700 mm (Gerhardt & Hytteborn 1992). Among the seasonally dry forests of South America, are the Neotropical seasonally dry forests (NSDFs), which are found in areas surrounding the Amazonian basin, in northeastern Brazil, eastern Paraguay and northeastern Argentina. Other, smaller, more isolated areas of NSDF occur in dry Andean valleys in Bolivia, Peru and Ecuador, in the dry Caribbean coast of Colombia and Venezuela, and are scattered through the Brazilian Cerrado biome on patches of fertile soils (Prado & Gibbs 1993). This forest is characterized by a typical flora (Pennington et al . 2000) among which Geoffroea spinosa Jacq. (Leguminosae) is particularly specific. Geoffroea spinosa has vertebrate dispersed fruits, and is found distributed in nearly all the areas described above. As part of a program aiming at analysing the postglacial distribution of NSDF typical species, micro- satellites have been developed in this species, together with another characteristic NSDF species, Astronium urundeuva (Caetano et al . 2005). DNA extract from a single individual from Peru was used, purified using Prep-A-Gene (Biorad) then digested using Alu I, Rsa I and Hae III (Boehringer). The digestion products were run on agarose gels and fragments from 500 to 1000 bp were selected. After a purification step, the frag- ments were ligated in pUC18/ Sma I/BAP following the manufacturer’s protocol (Amersham Pharmacia). Ultra- competent cells (Stratagene) were transformed using the ligation products, and white clones were then transferred onto nylon membranes. Clones were subsequently screened using (AC) 12 , (AG) 12 , (AAT) 8 and (TCT) 8 probes labelled with digoxigenin (DIG Oligonucleotide Tailing Kit, Roche Molecular). Hybridized colonies were detected using the DIG Luminescent Detection Kit (Roche Molecular) and used as polymerase chain reaction (PCR) templates for amplification using M13 forward and reverse universal primers. The PCR products were then sequenced using the same primers on an ABI377 PerkinElmer automated sequencer following the manufacturer’s protocol (Applied Biosystems). A total of 4300 recombinant clones were screened. Among the 53 positive clones found, 36 contained one to three microsatellites for a total of 41 different motifs. Two were Poly(T), with 12 ≤ n ≤ 13, 5 were of (AC) n type with 10 ≤ n ≤ 23, 14 were of (AG) n type with 8 ≤ n ≤ 34 and 2 of Correspondence: Y. Naciri Graven, Fax: + 4122 418 5101; E-mail: Yamama.Naciri-Graven@cjb.ville-ge.ch