Gradients of genetic diversity and differentiation across the distribution range of a Mediterranean coral: Patterns, processes and conservation implications

How historical and contemporary eco‐evolutionary processes shape the patterns of genetic diversity and differentiation across species’ distribution range remains an open question with strong conservation implications. Focusing on the orange stony coral, Astroides calycularis, we (a) characterized the pattern of neutral genetic diversity across the distribution range; (b) gave insights into the underlying processes; and (c) discussed conservation implications with emphasis on a national park located on a hotspot of genetic diversity.

Across species' distribution ranges, contemporary patterns of genetic diversity are modelled by the interplay among different eco-evolutionary processes such as genetic drift, gene flow, natural selection, survival and reproduction (e.g. Aurelle et al., 2011, Cahill et al., 2017Ledoux et al., 2010Ledoux et al., , 2020. These processes are deeply influenced by the spatial distribution and size of populations, which fluctuate over time in response to historical and contemporary biotic and abiotic factors. While negative gradients of neutral genetic diversity from central to peripheral populations were reported in various species (see Eckert et al., 2008;Hampe & Petit, 2005;Pironon et al., 2017), the underlying processes are still a matter of debate (Guo, 2012;Hardie & Hutchings, 2010). Two non-mutually exclusive hypotheses involving contemporary versus historical processes are usually considered to explain these negative gradients. Following the "central-peripheral hypothesis" (Sagarin & Gaines, 2002), the genetic diversity should decline from the centre towards the peripheries of a species' range in response to demo-genetic stochasticity (e.g. low effective size and low connectivity) linked to the environmental characteristics (e.g. spatial isolation and extreme environment) of peripheral habitats (e.g. Johansson et al., 2006). On the other hand, the "postglacial range expansion hypothesis" suggests that historical processes such as serial founder event recolonization (see Austerlitz et al., 1997Austerlitz et al., , 2000Slatkin & Excoffier, 2012) following the Last Glacial Maximum (LGM 24-18,000 years ago; Lambeck & Purcell, 2005) shape negative genetic gradients from the source to the edge of the expansion range. This pattern is mainly driven by an increase in genetic drift along the axis of expansion due to contrasted demographic histories when comparing peripheral versus central populations. Deciphering the relative impact of these processes on the current patterns of diversity is challenging, relying mainly on the evaluation of historical influences (Eckert et al., 2008;Guo, 2012).
Marine protected areas (MPAs) are an efficient tool to preserve genetic diversity and to mitigate the impacts of global change (Roberts et al., 2017). In many cases MPAs are designed following opportunistic rather than scientific criteria, leading to important mismatches between the protected area and the components of biological diversity (e.g. genetic diversity) to protect (Mouillot et al., 2011). With few exceptions (e.g. Dalongeville et al., 2018;Gazulla et al., 2021), genetic diversity and related processes are barely considered even though they are crucial for an effective functioning of the protected area (Palumbi, 2004). Among the key eco-evolutionary processes impacting the success of conservation strategies, connectivity within MPAs and among MPAs and neighbouring unprotected areas received particular attention (Magris et al., 2018;Manel et al., 2019). Indeed, the homogenizing effect of connectivity should counterbalance the disruptive effect of genetic drift induced by population depletion, allowing the maintenance and replenishment of genetic diversity. compared two estimators of pairwise genetic structure (G ST , D EST ) across the distribution range. The evolutionary and demographic history of the populations following the Last Glacial Maximum was reconstructed using approximate Bayesian computations and maximum-likelihood analyses. We inferred the contemporary connectivity among populations from Zembra National Park and with the neighbouring area of Cap Bon.

Results:
We demonstrate a decrease in genetic diversity and an increase in genetic differentiation from the Centre to the Eastern and Western Peripheries of the distribution range. Populations from Zembra show the highest genetic diversity reported in the species. We identified a spillover effect towards Cap Bon.

Main conclusions:
The patterns of genetic diversity and differentiation are most likely explained by "the postglacial range expansion hypothesis" rather than the "centralperipheral hypothesis." Enforcement of conservation measures should be considered to protect this genetic diversity pattern, in particular when considering the low effective population size inferred at many sites.

K E Y W O R D S
central-peripheral hypothesis, coral, genetic gradient, marine protected area, Mediterranean Sea, postglacial range expansion hypothesis program under grant agreement SEP-210597628 (FutureMARES)

Editor: Darren Yeo
The identification of populations in terms of their contribution to the connectivity of a network of populations is thus an important step to improve the management of MPAs (e.g. Gazulla et al., 2021;Lukoschek et al., 2016).
The Mediterranean orange stony coral, Astroides calycularis (Pallas, 1776), is an azooxanthellate scleractinian mainly found in rocky shallow habitats from the surface to about 40 to 60 m depth (Ocaña, 2012;Terrón-Sigler et al., 2016). While Quaternary fossils were recorded in the Northern Mediterranean (Zibrowius, 1980), the current established distribution range of A. calycularis is nearly linear, restricted in width to areas with minimum winter temperatures between 14 and 15°C (mean February temperature), extending from the neighbouring Atlantic Ocean to South Italy along the South-Western Mediterranean (Ocaña et al., 2015;Zibrowius, 1995; Figure 1). This coral is considered as warm-water species (Grubelić et al., 2004;Bianchi, 2007;Prada et al., 2019 but see Movilla et al., 2016), but some populations were recently reported in the eastern Adriatic and in Northern Italy (Bianchi, 2007;Kružić et al., 2002;Teixidó et al., 2020; Figure 1). Whether these atypical populations (i.e. outside of the currently accepted distribution range; Bianchi, 2007) result from species distribution expansion (Bianchi, 2007;Zibrowius, 1995) is still a matter of debate . Although it has been included in the annex II of the CITES and considered as vulnerable under Bern and Barcelona conventions (Templado et al., 2004), A. calycularis appears as least concern species in the IUCN red list due to the demographic stability of the populations and a relatively high abundance in the South-Western Mediterranean basin (Ocaña, 2012;Ocaña et al., 2009Ocaña et al., , 2015Otero et al., 2017).
Astroides calycularis is a gonochoric and brooding species characterized by internal fertilization occurring between April and May with larval release in June (Goffredo et al. 2010;Pellón & Badalamenti, 2016). A first assessment of the spatial pattern of genetic structure was conducted based on mitochondrial (COI) and nuclear (ITS) polymorphisms among 12 populations sampled in Western (i.e. Alboran Sea) and Eastern (i.e. Tyrrhenian Sea) Peripheries of the distribution range of the species (Figure 1). A slight genetic differentiation was observed between the two re- The established distribution range of Astroides calycularis (orange) is highly restricted in width to areas with minimum winter temperatures between 14 and 15°C (mean February temperature) (Ocaña et al. 2015;Zibrowius, 1995). Orange triangles correspond to atypical populations recently discovered (Bianchi, 2007;Kružić et al., 2002). The yellow star shows the hypothetical range distribution centre (DC). The four yellow lines correspond to the four range limits considered in this study: i) the Atlantic North-Western limit (ANW), the Atlantic South-Western limit (ASW), the Mediterranean North-Western limit (MNW) and the Mediterranean North-Eastern limit (MNE). Studied populations are shown in blue, green and pink and purple according to the results of the clustering analyses (see Results and using transcriptomic data (Teixidó et al., 2020). While these studies greatly improved our knowledge regarding the ecology of A.
calycularis, they were mainly focused on populations located at the Eastern and Western Peripheries. Whether those genetic patterns can be generalized to the Centre of the species range remains an open question with critical implication for the species' conservation.
The main objective of the study was to characterize the spatial pattern of genetic diversity in A. calycularis at two contrasted spatial scales. First, we considered most of the species distribution range and provided insights into the relative impact of contemporary ("central-peripheral hypothesis") versus historical ("postglacial range expansion hypothesis") processes on the pattern of genetic diversity.
Then, we deciphered the functioning of the Zembra National Park off the Tunisian coast, which harbour dense populations of A. calycularis (Boudouresque et al., 1986;Ghanem et al., 2019Ghanem et al., , 2021 implications of these results for the conservation of the species and for the management of the Zembra National Park.

| Sampling and datasets
Five to ten polyps of ten to 40 A. calycularis colonies from 13 popu-   Figure 1). These three populations were sampled in the same location among the two studies (Appendix S2).
The final dataset included 655 individuals from 29 populations genotyped with 12 microsatellites (Figure 1; Table 1). Considering the bathymetric range of the species and given that the sea level of the Mediterranean was 120m lower than it is today during the Last Glacial Maximum (24-18,000 years ago; Lambeck & Purcell, 2005), we considered that all these populations were recolonized since the LGM. Frequencies of null alleles were estimated for each locus and sample using in FREENA (Chapuis & Estoup, 2007). GENETIX 4.05 (Belkhir et al., 2004) was used to compute f, the Weir and Cockerham (1984) estimator of F IS , and to test for linkage equilibrium for each pair of loci overall populations and in each population, using 1,000 permutations.

| Spatial genetic structure
We performed a clustering analysis with STRUCTURE 2.2 (Pritchard et al., 2000) and a discriminant analysis of principal components (DAPC, Jombart et al., 2010) in ADEGENET (Jombart, 2008). The two analyses are described in Appendix S3.
Genotypic differentiation among populations was quantified using the Weir and Cockerham (1984) estimator of F ST , θ, and tested using an exact test (Raymond & Rousset, 1995) with default parameters in GENEPOP 4.7 (Rousset, 2008). Isolation-by-distance (IBD) pattern was analysed through the correlation of genetic and geographic distances among populations (Rousset, 1997) but considering only the 11 populations from Tunisia due to the sampling gap between the Western Periphery and Centre populations. The significance of the correlation between the genetic distances (F ST / (1-F ST )) and the logarithms of geographic distances (Ln(d)) was tested by the Mantel test with 10,000 permutations in GENEPOP.
Geographic distances were estimated following the most direct path among populations along the coastline in Google Earth (http:// earth.google.com). We estimated the "neighbourhood size" as the inverse of the slope of the linear regression between F ST /(1-F ST ) and Ln(d) (Rousset, 1997).

| Patterns of genetic diversity and structure over the distribution range accounting for geographic peripherality
We estimated the gene diversity, H e (Nei, 1973), in GENETIX 4.05 and computed the allelic richness (Ar (10) ) in ADZE (Szpiech et al., 2008) using the rarefaction method (Petit et al., 1998) with the minimum of 10 genes at a locus in a population. GESTE (Foll & Gaggiotti, 2006) was used to compute the population-specific F ST , as an estimate of the relative impact of genetic drift on the differentiation of the considered population (Gaggiotti & Foll, 2010). To characterize the pattern of genetic diversity and structure over the distribution range, we plotted H e , Ar (10) and the population-specific F ST s function of the longitude of the samples. Following a visual inspection of the data, we conducted a cubic regression to fit the curvilinear relationship among the variables (see Guo, 2012).
Bearing in mind the shape of the distribution range with multiple range limits (Figure 1), we complemented this approach following Yakimowski and Eckert (2007). We measured the rel- The second measure of geographic peripherality reflects the distance to the nearest current range limit (DNRL) following the coastline. Following Bianchi (2007), Ocaña et al. (2015) and Zibrowius (1995), we considered four range limits: the Atlantic North-Western limit (ANW; 36.515868°N; 6.292235°E), the Atlantic South- are at least two times higher than the distances estimated for the populations from the Eastern and Western Peripheries.
To characterize the spatial pattern of genetic diversity, we tested the significances of the slopes of the linear regressions among the two estimators of genetic diversity (H e and Ar (10) ), the estimate of genetic structure (population-specific F ST ) and the two measures of the geographic peripherality (DC and DNRL) using a permutation procedure (n = 10,000) in R (R Core Team, 2021).

| Comparison of the pairwise differentiation among populations from the Centre and the Peripheries of the distribution range
We used GENODIVE (Meirmans & vanTienderen, 2004) to compute and compare two measures of genetic differentiation quantifying complementary aspects of population structure (Jost et al., 2018): the nearness to fixation, G ST (Nei, 1987), and the relative degree of allelic differentiation, D EST (Jost, 2008 Here, the analysis was restricted to 0.55< D EST s <0.85. In each set, we fitted a multiple regression model including the "dataset" variable. We tested for the differences among the regression slopes of each category by looking at the significance of the D EST x "dataset" interaction.

| Influence of historical processes: evolutionary and demographic history
Following the postglacial range expansion hypothesis, centre to periphery negative gradients in genetic diversity (see Results) should result from an imprint of serial founder events. In this framework, the current peripheral populations are assumed to be farther from the ancestral population than the central populations.
We thus expect an increase in genetic drift along the recoloniza-  Table 2 and Appendix S4.

| Connectivity in Zembra National Park and neighbourhood populations
We inferred the connectivity among populations within the Zembra National Park and between the Park and the populations situated along the coast of Cap Bon. We conducted a filtered assignment analysis following Lukoschek et al. (2016) using Geneclass 2.0 (Piry et al., 2004). We conducted a first-generation migrant (FGM) analysis using the Bayesian criteria of Rannala and Mountain (1997) simulating 100,000 individuals and a type 1 error (alpha) of 0.005. Identified FGMs were removed from the dataset. In a second step, those FGMs were assigned to the reference dataset (i.e. without FGMs). In the last step, we considered a FGM to be assigned to a particular population when the assignment probability was higher than 0.01. Multiple assignments were allowed. When the assignment probability was lower than 0.01 for all populations, the migrant was considered as coming from an unsampled population.
F I G U R E 2 Five different scenarios were considered in the ABC analyses to reconstruct the evolutionary history of A. calycularis. N and t values correspond to population size and to timing in divergence events, respectively (time is not scaled). The retained scenario is surrounded in green. Posterior probability and corresponding 95% confidence interval of each scenario are shown For multiple tests, all significance levels were corrected using a false discovery rate (FDR) correction (Benjamini & Hochberg, 1995).

| RE SULTS
3.1 | Hardy-Weinberg equilibrium, genetic diversity and differentiation over the distribution range  (Table 3).
We demonstrated significant curve-linear relationships among H e , Ar (10) and population-specific F ST s and longitude (p <.01). This pattern was complemented by the significant decreases in H e , Ar (10) and increase in population-specific F ST s with the distance to centre (DC) and the significant increases in H e , Ar (10) and decrease in population-specific F ST s with the distance to the nearest range limit (DNRL) (Figure 3).
The first cluster encompassed all the individuals from the Centre     (Nei, 1973); f: Weir and Cockerham (1984) estimator of F IS (bold values are significant at 0.01); Ar (10) : rarefied allelic richness considering a minimum of 10 genes at a locus in a population; population-specific F ST and 95% confidence intervals. Distances to the nearest range limit and the hypothetical distribution centre are shown.

| Evolutionary history
The ABC analysis unambiguously supported Scenario V (posterior probability (95% CI) = 0.66 (0.65-0.67); see Figure 2) corresponding to a sequential foundation of the four different sub-clusters with a F I G U R E 3 Spatial pattern of genetic diversity: (a and b) Cubic regressions of H e , Ar (10) , the population-specific F ST and the longitude of the sampled locations following Guo (2012); (c and d) linear regressions of H e , Ar (10) , the populationspecific F ST function of the distance to the nearest range limit (DNRL, km; see Figure 1); (e and f) linear regressions of H e , Ar (10) , the population-specific F ST function of the distance to the centre range (DC, km; see Figure 1). H e , Ar (10) and the population-specific F ST are shown with circles, triangles and crosses, respectively. In plots ( Distance to Centre (km) He Ar (10) first divergence from the ancestral pool of the Southern East Italian sub-cluster (t 0 ), followed by a divergence of the Tunisian sub-cluster (t 1 ), and the Spanish (t 2 ) and the South Alboran 763-17,400) for the South Alboran sub-cluster to 15,500 (95% CI: 3,700-19,600) for the Spanish sub-cluster. Regarding the ancestral F I G U R E 4 Spatial genetic structure: (a) Clustering analysis conducted with STRUCTURE considering K = 2, K = 3 and K = 4. Each individual is represented by a vertical line partitioned in K-coloured segments, which represent the individual membership fraction in K clusters. Thin and thick black vertical lines delineate the different locations and regions, respectively. Samples names and geographic areas are shown below and above the assignment plots (for abbreviations, see Table 1). The mean membership coefficient for each cluster is shown. (b) Scatter plots of the discriminant analysis of principal components (DAPC) based on a a-score of 11. The left panel shows the plot corresponding to axes 1 and 2, and the right panel shows the plot corresponding to axes 1 and 3. The three axes represented 95.76% of the total variation in the data. Each dot corresponds to one individual (n = 655) from each of the 29 populations, which are represented by different colours. Inertia ellipses centre on the mean for each location and include 67% of the sampling points  Table 2. MIGRAINE analyses confirmed the relatively low effective population sizes suggested by the ABC analyses, with θ cur ranging from 0.47 (95% CI: 0.26-0.75) for PM to 4.8 (1.3-35) for DR (Table 4). Considering μ = 5*10 −4 (Sun et al., 2012) (Table 4).

| Connectivity in Zembra National Park and neighbouring of Cap Bon
The assignment tests indicated that 50 of the 198 individuals (25%) from Zembra and the neighbouring area of Cap Bon were considered as first-generation migrants (FGMs). FGMs were detected in all the 11 sites ranging from three (AIK, ZEM, NHS, MTS) to seven (CAS, LAM). Thirty-two FGMs (64%) could not be assigned to any sites (assignment probability <0.01) and were thus considered as migrants from unsampled sites. The remaining 18 FGMs (36%) were assigned with a probability higher than 0.01 to up to four sites. These putative sources only included five (AAA, AIK, CGD, ZEM and NHS) of the 11 sites. As we allowed for multiple assignments, these sites were identified as putative sources between one (NHS) and 13 (ZEM) times.
No FGM was assigned to the six other sites, which included four and two sites from Zembra and Cap Bon, respectively. None of the potential FGMs identified in Zembra were assigned to Cap Bon, but, on the opposite, four of the 11 FGMs identified in Cap Bon were assigned to Zembra.

| Whole distribution range pattern of genetic diversity and structure in A. calycularis
Knowledge regarding the spatial pattern of genetic diversity is critical to estimate populations' evolutionary potential and to support conservation planning Laikre et al., 2020). F I G U R E 5 Linear regression among pairwise G ST s (Nei, 1987; nearness to fixation) and D EST s (Jost, 2008; allelic differentiation) considering population pairs: (a) from the same periphery (in orange) or from the Centre (in red); (b) from the Western versus Eastern peripheries (in green), from the Centre versus Western periphery (in blue) and from the Centre versus Eastern periphery (in purple) Point estimates and 95% CI are shown for the scaled time (T g μ), the ancestral scaled population size (θ anc = 2*N anc *μ), the current scaled population size (θ cur = 2*N*μ) and the corresponding N ratio with μ corresponding to the mutation rate, N anc and N to the ancient and current population size, respectively. All the N ratio are significantly lower than 0 corresponding to a population bottleneck with the exception of DR and ECON (bold), which showed N ratio > 1 as expected in expanding populations and NREY (italics), which showed N ratio = 1 as expected in a stable population Periphery. Interestingly, nine of the ten highest pairwise D EST s (>0.79) involved one population from the Western Periphery and one population from the Eastern Periphery highlighting the totally different genetic pools of the populations located at the two peripheries.
Centre to peripheries negative genetic gradients were reported in different species (Eckert et al., 2008;Pironon et al., 2017) including corals (e.g. Andras et al., 2013;Miller & Ayre, 2004;Nunes et al., 2009). Focusing on the Mediterranean Sea, recent studies also reported regional negative genetic gradients (Boscari et al., 2019;Ledoux et al., 2018). Our work goes one step further as most of these studies focused on one particular area of the distribution range (Eckert et al., 2008;Guo, 2012). Indeed, considering most of the species distribution range, we demonstrate the occurrence of multidirectional (eastward and westward) longitudinal genetic gradients, involving different alleles, and going along with a stronger genetic differentiation and isolation of the peripheral populations.
Whether or not this pattern will be retained after adding Algerian

| "Postglacial range expansion hypothesis" versus "Central-peripheral hypothesis": insights into the underlying processes
Disentangling the relative influence of historical (i.e. postglacial sequential recolonization) versus contemporary (i.e. low effective population size, demographic isolation) processes on neutral genetic diversity gradient is a challenging task (Eckert et al., 2008; but see Johansson et al., 2006). Under the postglacial range expansion hy- Moreover, we call for complementary demographic studies (e.g. Ocaña et al., 2009;Goffredo et al., 2006;Prada et al., 2019) to formally characterize the impact of contemporary demo-genetic stochasticity on the pattern of genetic diversity (see Johansson et al., 2006).
An imprint of postglacial sequential recolonization on the contemporary pattern of genetic diversity was previously reported in the Mediterranean red gorgonian, Paramuricea clavata (Ledoux et al., 2018), and suspected in another coral, Leptosammia pruvoti, in the Adriatic Sea (Boscari et al., 2019). In spite of their relatively contrasted ecology (e.g. habitat, distribution range), these three species seem to show comparable genetic imprints of past climatic fluctuations. Similar legacy of postglacial recolonization processes is widely acknowledged in terrestrial species (e.g. Hewitt, 2000) and is expected in low dispersal marine species (Hellberg et al., 2002). Whether this trend can be generalized to other low dispersal Mediterranean habitat-forming species thus deserves further attention, owing to its conservation implications.

| General conservation implications and functioning of the Zembra National Park, a hotspot of genetic diversity
The need to account for genetic diversity is widely acknowledged by conservation biologists, yet it remains barely considered by policymakers Laikre et al., 2020). One of the main outputs of our study, supported by the approximate Bayesian computations and the maximum-likelihood approach, is the relatively low effective population sizes in A. calycularis. Considering μ = 5*10 -4 , most of the population size estimates are lower than 3,000 individuals with various populations (e.g. PM, GAT and ALG) showing values close to 500 individuals, the limit to consider a population as "genetically safe" (Jamieson & Allendorf, 2012). Included in Annex II of the CITES, A. calycularis is considered as "vulnerable" under Bern and Barcelona conventions and of "Least Concerned" in Mediterranean regional IUCN red list due to its demographic stability (Otero et al., 2017). Nevertheless, the low effective population sizes revealed here deserve further attention from policymakers and should be used to complement the conservation status of the species. Indeed, these low effective population sizes question the adaptive potential of the species in the current disturbance regime.
Besides, the occurrence of well-defined genetic clusters with con- likely to occur within the Zembra National Park (3.69 km 2 ). The assignment analyses showed that these populations are characterized by contrasted roles with regard to the connectivity of the system. The connectivity between Zembra and Cap Bon areas seems highly asymmetric. All the first-generation migrants (FGMs) shared between the two areas were found in Cap Bon and sourced in Zembra, suggesting spillover from the National Park towards the neighbouring area. This unidirectional pattern was expected considering the regional oceanographic features, and more particularly, the eastward Atlantic Tunisian Current (Sorgente et al., 2011). FGMs were identified in all the 11 Tunisian populations, but they only come from five populations. Those five source populations, and particularly ZEM and CDS, which were identified as potential sources in more than 70% of the cases, require specific conservation attention. A related point to consider here is the lack of assignment for 64% of the FGMs. The Zembra National Park is thus connected to some degree with unsampled area(s), likely on the western Tunisian coast considering the local oceanographic features (Sorgente et al., 2011). While it is important to concentrate management efforts on Zembra, the status of this hotspot of diversity seems unambiguously linked to neighbouring unprotected populations, reinforcing the relevance to enforce the protection at the species level.

| CON CLUS ION
In the warming Mediterranean Sea (Bensoussan et al., 2019;Cramer et al., 2018), the evolution of marine biodiversity is a matter of concern (Garrabou et al., 2009. Shifts in species distribution ranges are expected with "losers and winners" and potential consequences at the community level (Gómez-Gras et al., 2019, 2021. To date, most of the efforts in this topic focused on species negatively impacted by climate change (e.g. Arizmendi-Mejía, Ledoux, et al., 2015;Crisci et al., 2017;Ledoux et al., 2015;Torrents et al., 2008). Astroides calycularis may offer a complementary perspective. First, it is considered as a warm-water species, which may potentially benefit from the sea temperature increase expanding its distribution range (Bianchi, 2007;Kružić et al., 2002; but see Movilla et al., 2016). Then, addressing the impact of warming on biodiversity through the lens of peripheral populations is widely acknowledged (e.g. Hampe & Petit, 2005;Sexton et al., 2009). Accordingly, we identified in this study natural experimental setups where to concentrate efforts to decipher "peripheral population-by-warming environment" interactions.
Overall, these results combined with previous study Teixidó et al., 2020) stand A. calycularis as a highly relevant model to study the evolution of Mediterranean marine diversity facing warming.

ACK N OWLED G EM ENTS
Fieldwork and sampling were done in accordance with legislations.
The authors would like to acknowledge Hombre y Territorio (HyT) association and Dr. P. Casado de Amezúa for their collaboration.
Part of this work was carried out by using the resources of the na-

CO N FLI C T O F I NTE R E S T
None declared.

PEER R E V I E W
The peer review history for this article is available at https://publo ns.com/publo n/10.1111/ddi.13382.

DATA AVA I L A B I L I T Y S TAT E M E N T
Microsatellite genotypes are available on Dryad (DOI https://doi. org/10.5061/dryad.xgxd2 54gw).