Characterisation of thirteen polymorphic microsatellite markers for cowslip (Primula veris L.) developed using a 454 sequencing approach

Bickler, Charlotte; A’Hara, Stuart W.; Cottrell, Joan; Rogers, L. M.; Bridle, Jon R.

doi:10.1007/s12686-013-9997-2

Cited by 8 publications

(7 citation statements)

References 3 publications

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“…A total of 7615 unique reads contained at least one microsatellite motif, of which 4528 unique sequences were identified with sufficiently large flanking regions suitable for primer design (the sequences in these reads are available upon request). The microsatellite sequencing efficiency using Roche 454 high-throughput sequencing technology in P. poissonii (59.5%) was higher than that in Primula mistassinica (31.2%) [8] or Primula veris (22.2%) [16].…”

Section: Resultsmentioning

confidence: 99%

Characterization of Novel Microsatellite Loci for Primula poissonii (Primulaceae) Using High-Throughput Sequencing Technology

et al. 2016

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Primula poissonii (Primulaceae) is a perennial herb, widely distributed in the Hengduan Mountain region of Southwest China. In this study, Roche 454 pyrosequencing was used to isolate microsatellite markers. A total of 4528 unique sequences were identified from 68,070 unique reads. Of these, eighty-seven microsatellite loci were screened for utility using two criteria: successful PCR amplification and variation of these loci within three wild P. poissonii populations. Twenty loci were successfully amplified and exhibited polymorphic alleles. The number of observed alleles ranged from 1 to 9 with an average of 3.5. The observed and expected heterozygosities ranged from 0.087 to 1.000 and from 0.124 to 0.828, respectively. Among these SSR loci, only the P69 locus could not be cross-amplified successfully in two closely related species P. wilsonii and P. anisodora. The microsatellite loci developed in this study will be useful for studying genetic diversity and speciation events between P. poissonii and closely related Primula species.

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Section: Resultsmentioning

confidence: 99%

Characterization of Novel Microsatellite Loci for Primula poissonii (Primulaceae) Using High-Throughput Sequencing Technology

et al. 2016

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“…For genotyping, 12 primer pairs were used (developed by Bickler et al . 66 and Seino et al . 67 in 3 multiplex reactions for each locus containing 1 µl DNA sample, 5 µl of Qiagen Multiplex PCR Master Mix, 2 µl of one of the multiplexed primer combinations (1 µM for both the forward and reverse primer) and 2 µl of RNAse-free water.…”

Section: Methodsmentioning

confidence: 91%

Analysis of spatial genetic variation reveals genetic divergence among populations of Primula veris associated to contrasting habitats

Deschepper

Brys

Jacquemyn

2017

Sci Rep

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Genetic divergence by environment is a process whereby selection causes the formation of gene flow barriers between populations adapting to contrasting environments and is often considered to be the onset of speciation. Nevertheless, the extent to which genetic differentiation by environment on small spatial scales can be detected by means of neutral markers is still subject to debate. Previous research on the perennial herb Primula veris has shown that plants from grassland and forest habitats showed pronounced differences in phenology and flower morphology, suggesting limited gene flow between habitats. To test this hypothesis, we sampled 33 populations of P. veris consisting of forest and grassland patches and used clustering techniques and network analyses to identify sets of populations that are more connected to each other than to other sets of populations and estimated the timing of divergence. Our results showed that spatial genetic variation had a significantly modular structure and consisted of four well-defined modules that almost perfectly coincided with habitat features. Genetic divergence was estimated to have occurred about 114 generations ago, coinciding with historic major changes in the landscape. Overall, these results illustrate how populations adapting to different environments become structured genetically within landscapes on small spatial scales.

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“…Ten primer sequences for SSR markers previously developed for P. vulgaris, P. veris and P. elatior were identified from available literature (Van Geert et al 2006, Bickler et al 2013, Seino et al 2014, Supplementary material Appendix 4).…”

Section: Development Of Chloroplast Its and Ssr Markersmentioning

confidence: 99%

Recurrent hybridisation events between Primula vulgaris, P. veris and P. elatior (Primulaceae, Ericales) challenge the species boundaries: using molecular markers to re‐evaluate morphological identifications

Tendal

Ørgaard

Larsen

et al. 2018

Nordic Journal of Botany

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Three Primula species, Primula vulgaris, P. veris and P. elatior, have been objects of fascination for gardeners and botanists over several centuries. The species are able to hybridise, and where they co‐occur, hybrids are commonly found. In Denmark, Møns Klint on the island of Møn and Købelev Skov on Lolland are examples of localities where all three species occur and where the hybrids P. × digenea, the hybrid between P. vulgaris and P. elatior, and P. × polyantha, the hybrid between P. veris and P. vulgaris, can also be found. To investigate relationships between the species and their hybrids, we sampled 168 specimens from 10 geographical locations and subjected them to genetic analysis. The samples were also identified based on morphological traits: primarily inflorescense structure, the size, shape, colour and markings of corolla and leaf basis, leaf blade texture and hairiness. After identifying species‐specific SNPs in the Internal Transcribed Spacer sequence, these were used to resolve species and hybrid boundaries and status through a cleaved amplified polymorphic sequence assay. Polymorphisms in the chloroplast trnL sequence were used as a high‐throughput marker and used to determine the maternal parent of hybrids. Ten simple sequence repeat markers were applied to obtain further insight into the genetic makeup of the accessions using structure and Introgress, providing information of genetic variability within and between populations. Our results indicated that backcrossing of P. × digenea hybrids with parental species has occurred, and that many of the P. × digenea sampled were later‐generation hybrids rather than F1s. Analyses of P. × polyantha specimens show mostly the expected pattern for primary hybrids but indications of P. veris ancestry of a P. vulgaris plant was discovered. Our results further indicate that some of the specimens initially identified as P. elatior include P. vulgaris among their progenitors and thus challenge currently accepted species boundaries.

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Characterisation of thirteen polymorphic microsatellite markers for cowslip (Primula veris L.) developed using a 454 sequencing approach

Cited by 8 publications

References 3 publications

Characterization of Novel Microsatellite Loci for Primula poissonii (Primulaceae) Using High-Throughput Sequencing Technology

Characterization of Novel Microsatellite Loci for Primula poissonii (Primulaceae) Using High-Throughput Sequencing Technology

Analysis of spatial genetic variation reveals genetic divergence among populations of Primula veris associated to contrasting habitats

Recurrent hybridisation events between Primula vulgaris, P. veris and P. elatior (Primulaceae, Ericales) challenge the species boundaries: using molecular markers to re‐evaluate morphological identifications

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