Development of microsatellite markers in Fosterella rusbyi (Bromeliaceae) using 454 pyrosequencing

Abstract

The 15 SSR markers developed for F. rusbyi are promising candidates for population genetic analyses within F. rusbyi and other species of Fosterella.

Development of 15 nuclear microsatellite markers in Deuterocohnia (Pitcairnioideae; Bromeliaceae) using 454 pyrosequencing

“…From those sequences, we identified 835 perfect repeats with a minimum of seven units for di‐, six for tri‐, five for tetra‐, and four for penta‐ and hexanucleotide repeats, respectively. Sixty‐four microsatellite loci with sufficient flanking sequence and high repeat numbers were selected for PCR primer construction (Appendix 2), following previously described criteria (Wöhrmann et al., 2012). …”

“…(NiSch_06‐040F, NiSch_06‐040M), as well as in one individual each of D. seramisiana R. Vásquez, Ibisch & E. Gross (NiSch_06‐045K) and D. longipetala (NiSch_06‐068 as a positive control). PCRs were conducted in 12.5‐μL volumes in a T‐Gradient thermocycler (Biometra, Göttingen, Germany) following a touchdown protocol (Wöhrmann et al., 2012). As evidenced by electrophoresis on 1.5% agarose gels, 52 of the 64 primer pairs generated single, distinct PCR products within the expected size range in the positive control (Appendix 2).…”

“…Fragment sizes were scored with the help of an external size standard as described by Wöhrmann et al. (2012). …”

Reproduction biology and chloroplast inheritance in Bromeliaceae: a case study in Fosterella (Pitcairnioideae)

“…DNA was extracted from 100 mg of fresh leaves using a cetyltrimethylammonium bromide (CTAB) procedure (TelZur et al 1999;Krapp 2013). After initial primer tests, two EST-SSR markers derived from an expressed sequence tag (EST) database of Ananas comosus (Acom_12.12, Acom_101.1; Wöhrmann and Weising 2011) as well as two genomic SSR markers developed on the basis of 454 pyrosequences of Fosterella rusbyi accession JP06.0078 (ngFos_4, ngFos_11; Wöhrmann et al 2012) were selected for genotyping, based on the ability of these markers to clearly distinguish between the crossing parents of each treatment. PCR assays were performed as described previously (Wöhrmann and Weising 2011;Wöhrmann et al 2012).…”

“…Our objectives were (1) to study the preferred fertilization mode of these species (selfing or outcrossing), and (2) to analyze their ability to cross-hybridize with congeneric species from the same or a different evolutionary lineage. To achieve these objectives, offspring plants resulting from successful pollination were grown from seeds in the greenhouse, and genotyped along with their parents using a set of nuclear microsatellites (simple sequence repeats, hereafter 'nSSRs') that have been developed earlier in our group (Wöhrmann and Weising 2011;Wöhrmann et al 2012).…”