Precious coral species have been used to produce jewelry and ornaments since antiquity. Due to the high value and demand for corals, some coral beds have been heavily fished over past centuries. Fishing and international trade regulations were put in place to regulate fishing practices in recent decades. To this date, the control of precious coral exploitation and enforcement of trade rules have been somewhat impaired by the fact that different species of worked coral samples can be extremely difficult to distinguish, even for trained experts. Here, we developed methods to use DNA recovered from precious coral samples worked for jewelry to identify their species. We evaluated purity and quantity of DNA extracted using five different techniques. Then, a minimally invasive sampling protocol was tested, which allowed genetic analysis without compromising the value of the worked coral objects.The best performing DNA extraction technique applies decalcification of the skeletal material with EDTA in the presence of laurylsarcosyl and proteinase, and purification of the DNA with a commercial silica membrane. This method yielded pure DNA in all cases using 100 mg coral material and in over half of the cases when using “quasi non-destructive” sampling with sampled material amounts as low as 2.3 mg. Sequence data of the recovered DNA gave an indication that the range of precious coral species present in the trade is broader than previously anticipated.

Aim: Knowing a species' response to historical climate shifts helps understanding its perspectives under global warming. We infer the hitherto unresolved postglacial history of Pinus cembra. Using independent evidence from genetic structure and demographic inference of extant populations, and from palaeoecological findings, we derive putative refugia and re‐colonisation routes.Location: European Alps and Carpathians.Taxa: Pinus cembra. Methods: We genotyped nuclear and chloroplast microsatellite markers in nearly 3000 individuals from 147 locations across the entire natural range of P. cembra. Spatial genetic structure (Bayesian modelling) and demographic history (approximate Bayesian computation) were combined with palaeobotanical records (pollen, macrofossils) to infer putative refugial areas during the Last Glacial Maximum (LGM) and re‐colonisation of the current range.Results: We found distinct spatial genetic structure, despite low genetic differentiation even between the two disjunct mountain ranges. Nuclear markers revealed five genetic clusters aligned East–West across the range, while chloroplast haplotype distribution suggested nine clusters. Spatially congruent separation at both marker types highlighted two main genetic lineages in the East and West of the range. Demographic inference supported early separation of these lineages dating back to a previous interstadial or interglacial c. 210,000 years ago. Differentiation into five biologically meaningful genetic clusters likely established during postglacial re‐colonisation.Main Conclusions: Combining genetic and palaeoecological evidence suggests that P. cembra primarily survived the LGM in ‘cold period’ refugia south of the Central European Alps and near the Carpathians, from where it expanded during the Late Glacial into its current Holocene ‘warm period’ refugia. This colonisation history has led to the distinct East–West structure of five genetic clusters. The two main genetic lineages likely derived from ancient divergence during an interglacial or interstadial. The respective contact zone (Brenner line) matches a main biogeographical break in the European Alps also found in herbaceous alpine plant species.

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The production and trade of objects manufactured from the skeletal axis of coralid precious corals is a historically, culturally and economically important global industry. Coralids are members of the diverse Coralliidae family, which contains several species complexes and morphospecies. For most precious coral found in the jewelry trade, the color remains the sole clue and link to the taxonomic identity of the individual. Different coralid species have however similar or overlapping colors resulting in difficulty to taxonomically identify jewelry objects, including four species listed by the Convention on the International Trade of Endangered Species (CITES) whose international transport and trade requires species-specific and country of origin documentation. We aimed at developing a reliable method to taxonomically identify coralid material with the objective of distinguishing CITES protected species from their non-protected counterparts. We present Coral-ID, a genetic assay to taxonomically classify coralid objects using quasi non-destructive sampling. The assay classifies the analyzed sample in one of six taxonomic categories and performs at least presumptive separation of CITES-listed and non-listed species in all cases. Developmental validation experiments prove that Coral-ID is a specific, accurate and very sensitive method. As the first attempt to randomly sample corals in the trade to identify them, we applied Coral-ID on 20 precious coral objects seized by custom authorities upon import to in Switzerland. Thirteen (65%) of these samples could be analyzed; three of these were found to be presumptively CITES-listed, and 10 of them have proven to originate from non-CITES-listed species.

Precious corals from the Corallidae family (Corallium, Hemicorallium, and Pleurocorallium genera) are well known in the high-end jewelry industry due to their colorful and durable axial skeleton. They exist in various colors from white to pink to dark red. One highly appreciated shade is the light pink color, the so-called ‘angel skin’. This color is most often associated with Pleurocorallium elatius and Pleurocorallium secundum, species listed in CITES Appendix III. However, this has been based on an assumption of their visual similarity and has never been underpinned by detailed morphologic or genetic data. In this study, we present the analysis of an ‘angel skin’ coral necklace of exceptional size and homogeneous color and quality. Visual observation and Raman spectroscopy confirmed that the necklace consists of genuine, untreated precious coral material. Following minimally destructive sampling, respectively, drilling 2.2, 2.4, and 2.4 milligrams of material from the existing drill-holes, three randomly selected beads from the necklace were subject to a routine genetic identification assay, which is based on sequencing a short, taxonomically informative mitochondrial region. This genetic analysis identified the coral material as not from P. elatius or P. secundum but from another Pleurocorallium species. We subsequently sequenced additional mitochondrial DNA fragments from one ‘angel skin’ coral bead and compared them against a well-represented, curated reference data set of Pleurocorallium, including the first-ever sequencing of Pleurocorallium gotoense, Pleurocorallium johnsoni, Pleurocorallium cf. pusillum, and Pleurocorallium uchidai. We concluded that the analyzed material of the ‘angel skin’ coral necklace belongs to the Pleurocorallium norfolkicum species complex but is not identical to any hitherto analyzed and published Pleurocorallium specimens. A comparison with further taxonomically unidentified precious coral colony fragments identified a single sample fished in Vietnam to be completely identical to the ‘angel skin’ coral bead in the studied DNA regions. Thus, by the analysis of a high-end jewel, we discovered a species new to the jewelry trade and potentially also unknown to science. This implies that the currently considered list of species present in the precious coral trade is incomplete.

The holotype specimen holds the most authentic characteristics of a species; its features will serve as a foundation for the identification of individuals belonging to this species. The precious coral Pleurocorallium secundum was described in 1846 based on a colony from the Hawaiian Islands. This specimen has been preserved; however, it is decorticated and contains exclusively the axial skeleton, which has hindered its use for accurate species identification. Therefore, the species was redescribed in 1956 based on two lots collected in 1902. Pleurocorallium secundum was considered the most frequently fished precious coral species in the second half of the twentieth century with landings on the total scale of hundred-thousand kilograms, which was followed by its listing on CITES Appendix III. Recently, the conspecificity of the holotype and redescribed colonies was questioned, and specimens labeled in the scientific literature as P. secundum were discovered to be phylogenetically distant from each other. To clarify the identity of P. secundum, we took minimally destructive samples from the century-old holotype and fragments of redescribed colonies and applied techniques conforming to low copy number DNA analyses. DNA sequences of three mitochondrial regions were evaluated in a phylogenetic framework together with DNA sequences retrieved from freshly collected putative P. secundum specimens and sequences from the scientific literature. The results of this study clearly indicate that the holotype and the colony fragments used for the redescription of P. secundum represent the same species. Based on the specimens confirmed to be P. secundum with genetic evidence, the distribution area of P. secundum stretches from the Hawaiian Islands to the South China Sea. At the same time, our analysis uncovered both published and fresh specimens that are in fact not P. secundum. The latter includes the fished Makapuu coral bed in Hawaii, which used to be a significant coral fishing area. Based on the microscopic analysis of redescribed colony fragments, we complement the diagnosis of P. secundum. This, together with our genetic results will aid the identification of coral objects present in the international jewelry trade by providing authentic molecular barcoding markers and morphologic features for the identification of P. secundum.