Novel microsatellite markers for evaluation of genetic diversity in the tetraploid flame azalea, Rhododendron calendulac
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Novel microsatellite markers for evaluation of genetic diversity in the tetraploid flame azalea, Rhododendron calendulaceum (Ericaceae) Megan L. Thompson1 · Zack E. Murrell2 · Charles N. Horn3 · Kathleen A. Kron4 · Emily L. Gillespie5 Received: 8 July 2020 / Accepted: 4 September 2020 © Springer Nature B.V. 2020
Abstract Microsatellite markers have become a popular and useful tool for investigating evolutionary processes at shallow taxonomic scales such as within a species or between extremely closely related species. Rhododendron sect. Pentanthera is a closely related group of deciduous azaleas that demonstrate both naturally occurring and horticulturally derived hybridization. Two species, flame azalea and Cumberland azalea, represent a particularly recalcitrant evolutionary problem, which will benefit from the development of rapidly evolving molecular markers. Microsatellite markers were specifically developed for Rhododendron calendulaceum, the flame azalea, for use in studies of genetic structure and potential hybridization with its close relative Rhododendron cumberlandense, the Cumberland azalea. Forty-eight primer pairs designed from paired-end Illumina MiSeq data were screened for robust amplification. Sixteen of these pairs were PCR-amplified in the presence of fluorescently labeled primers and genotyped in 66 flame azalea individuals from three geographically dispersed populations. Fifteen primer pairs were both reliable and polymorphic and exhibit ample variability for use in downstream populationlevel investigations. Cross-amplification in all other members of Rhododendron sect. Pentanthera was highly successful, suggesting broad utility across the entire clade. The novel microsatellite markers presented here functioned well within the target species and amplified with high success in the remaining members of the clade. They represent a significant improvement to the genetic toolkit available for Rhododendron sect. Pentanthera, and particularly for the flame/Cumberland azalea evolutionary problem. Keywords Ericaceae · Microsatellite markers · Rhododendron calendulaceum · Rhododendron cumberlandense · Rhododendron subsect. Pentanthera · Southeastern United States
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05813-9) contains supplementary material, which is available to authorized users.
Rhododendron sect. Pentanthera comprises 17 species of morphologically similar, closely related azaleas, which collectively exhibit a tertiary distribution. One species (Rhododendron molle (Blume) G. Don) is found in China and Japan.
* Emily L. Gillespie [email protected]
1
DeBusk College of Osteopathic Medicine, Lincoln Memorial University, Harrogate, TN, USA
Megan L. Thompson [email protected]
2
Department of Biology, Appalachian State University, Boone, NC, USA
Zack E. Murrell [email protected]
3
Department of Sciences and Mathematics, Newberry College, Newberry, SC, USA
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