Development of microsatellite markers in Moldenhawera blanchetiana and their transferability to M . luschnathiana (Fabac
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Development of microsatellite markers in Moldenhawera blanchetiana and their transferability to M. luschnathiana (Fabaceae) Caio Vinicius Vivas • Ca´ssio van den Berg • Luciano Paganucci de Queiroz • Fernanda Amato Gaiotto
Received: 9 September 2014 / Accepted: 27 September 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract The first microsatellite (SSR) markers were developed for Moldenhawera blanchetiana and transferred to M. luschnathiana, two species endemic to the Atlantic Forest, Brazil. Sixteen loci for M. blanchetiana were characterized, 15 of which showed high levels of heterozygosity (HE = 0.645 and HO = 0.559). The fifteen loci were transferred to M. luschnathiana where 14 were polymorphic (HE = 0.567 and HO = 0.550). The average numbers of alleles per locus was 6.6 for M. blanchetiana and 4.5 for M. luschnathiana. High values of the combined exclusion probability (Q [ 0.999, for both species) demonstrated the potential of these loci for paternity testing. These microsatellite markers are useful tools for performing genetic and evolutionary studies as well as for the management and conservation of populations of these two species. Keywords SSR Conservation genetics Transferability NGS Atlantic forest Moldenhawera is a genus of Fabaceae that is restricted to eastern Brazil. Ten species and 11 taxa are recognized, distributed in the Atlantic Forest, Cerrado, and Caatinga Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0351-0) contains supplementary material, which is available to authorized users. C. V. Vivas F. A. Gaiotto (&) Departamento de Cieˆncias Biolo´gicas, Universidade Estadual de Santa Cruz, Rodovia Ilhe´us-Itabuna Km 16, Salobrinho, Ilhe´us, BA 45662-900, Brazil e-mail: [email protected] C. van den Berg L. P. de Queiroz Departamento de Cieˆncias Biolo´gicas, Universidade Estadual de Feira de Santana, Feira de Santana, BA, Brazil
phytogeographic domains in areas under anthropogenic disturbances (Queiroz et al. 1999). Moldenhawera blanchetiana Tul. and M. luschnathiana Yakovlev are tree species endemic to the rain forests of the Atlantic Forest in the state of Bahia, Brazil. Here, we report the development of 16 microsatellite (simple sequence repeat, SSR) markers for M. blanchetiana and transferability of 15 of these loci in M. luschnathiana, which will allow genetic and evolutionary studies as well as assist in designing conservation strategies for these species in the coastal Atlantic forests. We extracted DNA from a single M. blanchetiana individual (voucher Vivas, 230 HUEFS) using the cetyltrimethylammonium bromide (CTAB 2 %) method, for genomic library construction. The library was enriched with (AC)n and (AG)n DNA probes and sequenced using 454 pyrosequencing technology (Ecogenics GmbH, Schlieren, Switzerland). Thirty primers pairs were tested, but only 16 were successfully amplified in M. blanchetiana (n = 28), and 15 of them were transferred to M. luschnathiana (n = 20) (Table 1). The PCR reactio
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