Anonymous single‐copy nuclear DNA (scnDNA) markers for two endemic log‐dwelling beetles: Apasis puncticeps and Adelium calosomoides (Tenebrionidae: Lagriinae: Adeliini)

Abstract

Three approaches — microsatellite library screening, consensus primer PCR (polymerase chain reaction) and sequencing with arbitrary primer pairs (SWAPP) — were used to develop single‐copy nuclear DNA (scnDNA) markers for log‐dwelling beetles Apasis puncticeps and Adelium calosomoides. We are unaware of other nuclear markers for Adeliini. We tested > 70 primer pairs per species, but despite exhaustive optimization, we obtained only five polymorphic markers. Nonetheless, the markers are valuable in detection of effects of habitat fragmentation.

When log‐dwellers meet loggers: impacts of forest fragmentation on two endemic log‐dwelling beetles in southeastern Australia

“…in press), we excluded these loci from this species’ data set when examining possible impacts of the three predictor variables (above) on F IS , and for species comparisons of F IS . As explained in Schmuki et al . (in press), continuous and fragmented forest sites were equally deficient in heterozygotes at A D 19 and A D 32; we thus found it legitimate to include these loci when comparing the two in other analyses.…”

“…puncticeps were genotyped at single‐copy nuclear (scn) DNA loci AP38, A1Mp20U2, S2G m 49 and all Ad. calosomoides at A D 19 and A D 32 as per Schmuki et al . (in press).…”

“…To avoid patterns of F IS for Ad. calosomoides being driven by homozygous excess at A D 19 and A D 32 (Schmuki et al . in press), we excluded these loci from this species’ data set when examining possible impacts of the three predictor variables (above) on F IS , and for species comparisons of F IS .…”

Beyond Roadkill, Radiotracking, Recapture and FST—a Review of Some Genetic Methods to Improve Understanding of the Influence of Roads on Wildlife

“…Multiple variable markers may not be available for a given target species, in which case marker development may be necessary. This can be challenging for certain taxa (e.g., Tenebrionid beetles, Schmuki et al 2006a) and costly. However, even the very effective modern methods for developing microsatellite markers (e. g., Edwards et al 1996) are rapidly being supplanted by inexpensive "next-generation" genome sequencing (Ellegren 2008, Abdelkrin et al 2009).…”

Development and application of three-tiered nuclear genetic markers for basal Hexapods using single-stranded conformation polymorphism coupled with targeted DNA sequencing

“…However, isolation of microsatellites from certain invertebrate groups is notoriously difficult (e.g. Onychophora, [ 16 ]; Lepidoptera, [ 17 , 18 ]; tenebrionid beetles, [ 19 ]), and once obtained, non-amplifying 'null' alleles, size homoplasy, and low levels of polymorphism can severely undermine their utility. For example, after considerable effort, five of 11 microsatellite loci developed by van der Wurff [ 20 ] for the Collembolon Orchesella cincta were essentially unusable, and three of the remaining six loci had four or fewer alleles from 376 individuals assayed [ 21 ].…”