Microsatellite In Aeschynomene Falcata (leguminosae): Diversity, Cross-amplification, And Chromosome Localization

Aeschynomene falcata is an important forage species; however, because of low seed production, it is underutilized as forage species. Aeschynomene is a polyphyletic genus with a challenging taxonomic position. Two subgenera have been proposed, and it is suggested that Aeschynomene can be split in 2 genera. Thus, new markers, such as microsatellite sequences, are desirable for improving breeding programs for A. falcata. Based on transferability and in situ localization, these microsatellite sequences can be applied as chromosome markers in the genus Aeschynomene and closely related genera. Here, we report the first microsatellite library developed for this genus; 11 microsatellites were characterized, with observed and expected heterozygosities ranging from 0.0000 to 0.7143 and from 0.1287 to 0.8360, respectively. Polymorphic information content varied from 0.1167 to 0.7786. The departure from Hardy-Weinberg equilibrium may have resulted from frequent autogamy, which is characteristic of A. falcata. Of the 11 microsatellites, 9 loci were cross-amplified in A. brevipes and A. paniculata and 7 in Dalbergia nigra and Machaerium vestitum. Five of these 7 cross-amplified microsatellites were applied as probes during the in situ hybridization assay and 2 showed clear signals on A. falcata chromosomes, ensuring their viability as chromosome markers.1341039010397Begum, R., Alam, S.S., Menzel, G., Schmidt, T., Comparative molecular cytogenetics of major repetitive sequence families of three Dendrobium species (Orchidaceae) from Bangladesh (2009) Ann. Bot, 104, pp. 863-872Billotte, N., Lagoda, P.J.R., Risterucci, A.M., Baurens, F.C., Microsatellite-enriched libraries: Applied methodology for the development of SSR markers in tropical crops (1999) Fruits, 54, pp. 277-288Cardoso, D., de Queiroz, L.P., Pennington, R.T., de Lima, H.C., Revisiting the phylogeny of papilionoid legumes: New insights from comprehensively sampled early-branching lineages (2012) Am. J. Bot, 99, pp. 1991-2013Creste, S., Tulmann Neto, A., Figueira, A., Detection of single sequence repeat polymorphisms in denaturing polyacrylamide sequencing gels by silver staining (2001) Plant Mol. Biol. Rep, 19, pp. 299-306Ellis, J.R., Burke, J.M., EST-SSRs as a resource for population genetic analyses (2007) Heredity, 99, pp. 125-132Flajoulot, S., Ronfort, J., Baudouin, P., Barre, P., Genetic diversity among alfalfa (Medicago sativa) cultivars coming from a breeding program, using SSR markers (2005) Theor. Appl. Genet, 111, pp. 1420-1429Jones, R.M., McDonald, C.K., Clements, R.J., Bunch, G.A., Sown pastures in subcostal south-eastern Queensland: Pasture composition, legume persistence and cattle live weight gain over 10 years (2000) Trop. Grassl, 34, pp. 21-37Kalia, R.K., Rai, M.K., Kalia, S., Singh, R., Microsatellite markers: An overview of the recent progress in plants (2011) Euphytica, 177, pp. 309-334Kemp, S., (2002) PIC Calculator Extra, , http://www.genomics.liv.ac.uk/animal/pic.html, Accessed September 4, 2012Klitgaard, B.B., Lavin, M., Tribe Dalbergieae Sens. Lat (2005) Legumes of the World, pp. 307-335. , (Lewis GP, Schrire BD, MacKinder BA and Lock M, eds.). Royal Botanic Gardens, KewMiller, M.P., (1997) Tools for Populations Genetics Analyses V 1. 3. A Windows® program for the analysis of allozyme and molecular population genetic data, , http://www.ccg.unam.mx/~vinuesa/tlem09/docs/TFPGADOC.PDF, Accessed April 4, 2014Moraes, A.P., Leitch, I.J., Leitch, A.R., Chromosome studies in Orchidaceae: Karyotype divergence in Neotropical genera in subtribe Maxillariinae (2012) Bot. J. Linn. Soc, 170, pp. 29-39Pinheiro, F., Palma-Silva, C., de, B.F., Cozzolino, S., Cross-amplification and characterization of microsatellite loci for the Neotropical orchid genus Epidendrum (2009) Genet. Mol. Biol, 32, pp. 337-339Polido, C., (2013) Estudo Cromossômico da Tribo Dalbergieae Sensu Klitgaard & Lavin (2005), com Ênfase no Clado Dalbergia Sensu Stricto (Leguminosae, Papilionoideae), , Doctoral thesis, Universidade Estadual de Campinas, CampinasQueiroz, L., Cardoso, D.B.O.S., A new species of Aeschynomene L. (Leguminosae, Papilionoideae) from a continental sand dune area in north-eastern Brazil (2008) Bot. J. Linn. Soc, 157, pp. 749-753Ribeiro, R.A., Lavin, M., Lemos-Filho, J.P., Mendonça-Filho, C.V., The genus Machaerium (Leguminosae) is more closely related to Aeschynomene sect. Ochopodium than to Dalbergia: Inferences from combined sequence data (2007) Syst. Bot, 32, pp. 762-771Rousset, F., Genepop'007: A complete re-implementation of the genepop software for Windows and Linux (2008) Mol. Ecol. Resour, 8, pp. 103-106Ruiz, C., Asins, M.J., Comparison between Poncirus and Citrus genetic linkage maps (2003) Theor. Appl. Genet, 106, pp. 826-836Schwarzacher, T.P., Heslop-Harrison, P., (2000) Practical in Situ Hybridization, , 1st edn. Bios, OxfordTemnykh, S., DeClerck, G., Lukashova, A., Lipovich, L., Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): Frequency, length variation, transposon associations, and genetic marker potential (2001) Genome Res, 11, pp. 1441-1452Thiers, B., (2014) Index Herbariorum: A global Directory of Public Herbaria and Associated Staff, , http://sciweb.nybg.org/science2/IndexHerbariorum.asp, New York Botanical Garden's Virtual Herbarium. Available at. Accessed April 4, 201