Development of Anonymous Nuclear Loci for Pterospora andromedea (Monotropoideae) Using Illumina and Ion Torrent Sequenci
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TECHNICAL NOTE
Development of Anonymous Nuclear Loci for Pterospora andromedea (Monotropoideae) Using Illumina and Ion Torrent Sequencing Data N. J. Dowie1 · L. C. Grubisha2 · B. A. Burton1 · M. R. Klooster3 · S. L. Miller1
Received: 7 September 2016 / Accepted: 17 January 2017 © Springer Science+Business Media Dordrecht 2017
Abstract The mycoheterotrophic plant Pterospora andromedea is an important species in understanding the evolution and ecology of highly specialized plant-fungal symbioses. Lineages within P. andromedea have been shown to engage in highly specific interactions with obligate fungal associates found in Rhizopogon subgenus Amylopogon. Unfortunately, evidence of highly specific interactions resulted from single locus analyses and this result requires further examination. If highly specific host-symbiont selectivity is present, then the lack of compatible partners may best explain the conservation status of P. andromedea. To better evaluate and support conservation efforts for P. andromedea, more molecular loci are necessary. 115 candidate loci were screened from Illumnia MiSeq and Ion Torrent next-generation sequencing and resulted in the development of four polymorphic anonymous nuclear loci. These loci will provide the means to critically examine host-symbiont specificity for P. andromedea lineages across the endemic range and will assist conservation initiatives. Keywords Pterospora andromedea · Rhizopogon · Amylopogon · Anonymous Nuclear Loci · ANL · Conservation · Monotropoideae · Ericaceae * N. J. Dowie [email protected] 1
Department of Botany, University of Wyoming, 1000 University Avenue, Laramie, WY 82071, USA
2
Department of Natural and Applied Sciences, University of Wisconsin - Green Bay, 2420 Nicolet Drive, Green Bay, WI 54311, USA
3
Centre College, 600 West Walnut Street, Young Hall 243, Danville, KY 40422, USA
Pterospora andromedea, Ericaceae, is an ideal candidate to explore host-symbiont specificity in a mycoheterotrophic plant across a geographic mosaic. Mycoheterotrophs are plants of high ecological interest as they obligately associate with fungal partners and may subsequently have increased sensitivity to environmental perturbations (Merckx et al. 2013). The investigation of P. andromedea is particularly worthwhile since it is entirely nonphotosynthetic and relies on persistent, exclusive, and specific symbioses with species in Rhizopogon subgenus Amylopogon (Bidartondo and Bruns 2002; Dowie et al. 2016). Evidence exists for plant lineage specificity with fungal species under sympatric conditions that represent a mosaic of symbiont specificity across North America (Bidartondo and Bruns 2002; Dowie et al. 2016; Grubisha et al. 2014a). The lack of effective specific symbionts has been suggested as a primary reason for the threatened conservation status of P. andromedea in many areas (Grubisha et al. 2014a; Hazard et al. 2012). Unfortunately, the traditional molecular locus used for identifying lineages of P. andromedea, plastid pseudogene trnL, is unalignable wi
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