A collection of 5,659 expressed sequence tags (ESTs) from pineapple [Ananas comosus (L.) Merr.] was screened for simple sequence repeats (EST-SSRs) with motif lengths between 1 and 6 bp. Lower thresholds of 15, 7 and 5 repeat units were used to define microsatellites of the mono-, di-, and tri- to hexanucleotide repeat type, respectively. Based on these criteria, 696 SSRs were identified among 3,389 EST unigenes, together representing 2,840 kb. This corresponds to an average density of one SSR every 4.1 kb of non-redundant EST sequences. Dinucleotide repeats were most abundant (38.4% of all SSRs) followed by trinucleotide repeats (38.1%). Flanking primer pairs were designed for 537 EST-SSR loci, and 49 of these were screened for their functionality in 12 accessions of A. comosus, 14 accessions of 5 additional Ananas species and 1 species of Pseudananas. Distinct PCR products of the expected size range were obtained with 36 primer pairs. Eighteen loci analyzed in more detail were all polymorphic in pineapple, and primer pairs flanking these loci also generated PCR products from a wide range of genera and species from six subfamilies of the Bromeliaceae. The potential to reveal polymorphism in a heterologous target species was demonstrated in Deuterocohnia brevifolia (subfamily Pitcairnioideae).
The strong genetic differentiation between southeastern Caatinga and the remaining regions may indicate the occurrence of a cryptic species in . The unique genetic composition of each inselberg population suggests in situ conservation as the most appropriate protection measure for this plant lineage.
We applied a series of intra-and interspecific in situ cross-pollination experiments under greenhouse conditions to evaluate the breeding systems in four Fosterella species (Pitcairnioideae s.str.; Bromeliaceae). Viable hybrids were produced between each pair of the investigated species, suggesting that reproduction barriers may be low also under natural conditions. Seed germination rates proved to be high in each crossing treatment, indicating a high viability of the artificial hybrids. Large numbers of seeds were produced after both closed and open pollination treatments, suggesting that autogamy may be a major reproductive strategy in the genus. Our results support the concept that self-compatibility is an appropriate way to avoid natural hybridization in Bromeliaceae and could assist in maintaining species integrity in the presence of pollen flow. Paternity was verified in all crosses by genotyping parents and offspring with a set of polymorphic nuclear microsatellite markers. To study the mode of chloroplast inheritance, we developed a novel set of 24 chloroplast microsatellite markers using 454 pyrosequencing technology, and applied four of these markers for genotyping parents and offspring from all crosses. Our results clearly demonstrated a maternal inheritance of plastids.
A population genetic analysis based on eight genomic SSR markers and three EST-SSR (expressed sequence tags) markers developed in peach (Prunus persica (L.) Batsch) and Japanese plum (Prunus salicina Lindl.) was carried out in 12 wild populations of cherry plum (Prunus divaricata Ledeb.) sampled along the Iranian coast of the Caspian Sea. A total of 184 alleles (3-31 per locus) were detected with a mean value of 16.7 alleles per locus. None of the loci or populations showed deviation from Hardy-Weinberg equilibrium, and all markers proved to be unlinked. The mean values for the observed and the expected heterozygosity were 0.66 and 0.73, respectively. There was very little genetic differentiation among populations, as was indicated by low overall values of Wright's F ST (0.03) and Nei's G ST (0.08). An analysis of molecular variance (AMOVA) showed that 96.8% of the total variance was attributable to differences between individuals within populations. Genetic and geographic distances were nevertheless positively correlated, as evidenced by a Mantel test. The high level of genetic diversity and the apparent lack of genetic structure in wild P. divaricata may be attributed to frequent long distance gene flow through frugivorous birds and possibly humans, as has been documented for other Prunus species.
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