Genetic diversity and differentiation of two narrowly endemic Origanum Species in Morocco: implications for conservation

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Classification within the Origanum genus is complicated by extensive morphological variation and frequent interspecies hybridization. In this study, we assessed the genetic diversity and population structure of O. elongatum and O. grosii across their full distribution in Morocco, sampling 264 individuals from seven populations and analyzing them with 13 microsatellite (SSR) markers. Despite its restricted native range, O. elongatum displayed considerable genetic diversity (Ho = 0.455, He = 0.429), whereas O. grosii showed lower diversity (Ho = 0.285) but higher differentiation among populations. Bayesian clustering, Principal Coordinates Analysis, and STRUCTURE analysis revealed four main genetic groups largely corresponding to geographic origin and clearly separating the two species. These results indicate strong species divergence and the existence of distinct population structures, underscoring the need for species-specific conservation measures. We recommend integrating in-situ and ex-situ strategies, including habitat protection, genetic reinforcement, and germplasm preservation in gene banks. Overall, this study illustrates how genetic data can inform conservation planning for rare endemic plants in ecologically sensitive regions, providing a model applicable to other Mediterranean mountain ecosystems facing climate and land-use pressures. Conservation Genetics Endemic species Origanum elongatum Origanum grosii Genetic Diversity SSR Markers Figures Figure 1 Figure 2 Figure 3 Introduction Morocco is a major biodiversity hotspot in the Mediterranean, hosting about 3,912 taxa and 1,298 subspecies, with roughly 22% endemism (Fennane and Ibn Tattou 2012 ). This exceptional endemic richness (878 taxa, nearly 600 at the species level) is mainly concentrated in the High Atlas, Middle Atlas, and Rif mountains. However, this diversity is increasingly threatened, as nearly 2,193 plant species are endangered, over half of which (53%) are highly vulnerable due to anthropogenic pressures and environmental change (Fennane and Ibn Tattou 1999 ). Endemics with narrow distributions are particularly at risk, as their limited range increases their vulnerability to stochastic events and environmental stressors (George et al. 2009 ; Lannuzel et al. 2021 ). The genus Origanum L. (Lamiaceae), comprising about 18 hybrids and 43 species, is largely distributed around the Mediterranean region (Ietswaart, 1980 ). In Morocco, four taxa and one hybrid are recognized: O. compactum Benth. (also found in southern Spain), O. vulgare subsp. virens , and three endemic taxa : O. elongatum , O. grosii , and O. × font-queri (a spontaneously occurring hybrid resulting from O. grosii and O. compactum ) (Bakha et al. 2017 ). The species O. elongatum has a geographic range that stretches mainly across northeastern Morocco, between the Middle Atlas and Rif Mountain systems, typically at high elevations (Bakha et al. 2020 ). O. grosii is among the rarest species, occurring in small, isolated populations in the humid western Rif at high altitudes (Bakha et al. 2017 ). Taxonomic delimitation within Moroccan Origanum is controversial. Morphological similarities among O. grosii , O. elongatum and the hybrid O. × font-queri have led to confusion. Although certain databases (Euro + Med 2006 ; African Plant Database v3.4.0, Dobignard and Chatelain 2010–2013) classify O. grosii and the hybrid O. × font-queri under the same taxon as O. elongatum (Bakha et al. 2017 ), Ietswaart ( 1980 ) designated O. grosii as an independent species, based on morphological traits, including its shorter stems and leaves, as well as broader bracts in comparison to O. elongatum . Populations of Moroccan Origanum species are highly fragmented and increasingly threatened by drought, climate change, habitat loss, overgrazing, and deforestation (Aboukhalid et al. 2017 ). Such fragmentation and population decline often result in enhanced genetic drift, reduced gene flow, and decline in genetic variability, all of which compromise the adaptive potential and long-term persistence of species (Holsinger and Gottlieb 1991 ). Low levels of genetic variability and high levels of genetic divergence among populations are common characteristics of rare and geographically isolated species (Sullivan et al. 2019 ). Endemic medicinal plants like Origanum species are highly vulnerable to habitat degradation and climate change, emphasizing the need for conservation (Aboukhalid et al. 2017 , 2023 ). Genetic diversity is crucial for adaptive capacity and long-term survival (Reed and Frankham 2003 ; Supple and Shapiro 2018 ), yet small populations often lose diversity through drift, bottlenecks, and founder effects (Banks et al. 2015 ), making its conservation a central goal (Frankham 1996 ). This study addresses the previously unexamined genetic diversity and population structure of Moroccan endemic O. elongatum and O. grosii using genomic and landscape approaches across their natural range. In particular, we investigate the following key questions: (i) What are the patterns of genetic polymorphism and population structure in O. elongatum and O. grosii across and within populations? (ii) Is there evidence of gene flow among these species? (iii) Do patterns of genetic differentiation support the current taxonomic distinctions proposed by Ietswaart ( 1980 )? (iv) How might the genetic results contribute to the development of effective conservation and management plans for these rare and endangered endemics? Materials and Methods Study Species Origanum elongatum Emberger & Maire (Lamiaceae) is a perennial species native exclusively to Morocco. It is naturally distributed in Rif Mountain systems and the northern Middle Atlas, predominantly inhabiting schistose soils at elevations ranging from 681 to 1866 meters, with most populations occurring above 1300 m (Bakha et al. 2020). O. elongatum is a climate-tolerant species found from semi-arid to per-humid zones and is valued in Morocco for its aromatic, culinary, melliferous, and ornamental uses. It is charachterized by tall purplish stems reaching up to 90 cm. Branches are largely composed of terminal spikes. Leaves are arranged in up to 30 opposite pairs per stem, becoming subsessile toward the apex, with petioles up to 5 mm in length (Ietswaart 1980). The flowering occurs from June to October. The leaves are densely encrusted with glandular trichomes that produce essential oils, resulting in a fragrant and flavored odor characteristically associated with this species (Simon et al. 1984). Although specific pollination studies are lacking, flower morphology, strong scent, prominent stamens (up to 3.5 mm), and elongated styles (up to 8 mm), as well as the frequent visits by pollinating insects suggest a predominance of cross-pollination. This is consistent with the general reproductive patterns reported in the Lamiaceae family, which is typically insect-pollinated and predominantly outcrossing (Judd et al. 1999). Origanum grosii Pau & Font Quer ex Ietswaart stands out as one of the scarcest and most localized species taxonomically assigned to the genus Origanum . This species is found only in the western Rif Mountains of northwestern Morocco. It grows mainly at high elevations under humid conditions, typically on calcareous substrates, at elevations ranging between 400 and 1900 m (Bakha et al. 2017). Morphologically, O. grosii is quite distinctive. It has relatively short stems (up to 55 cm), which are lightly hairy, and long lateral branches stretching between 4 and 30 cm. Its bracts are notably larger (2–5 mm) than those of related species. The leaves are smaller than those of O. elongatum , usually between 5 and 19 mm long. The species has erected purplish-brown stems, with fine hairs (trichomes) concentrated at the base and continuing sparsely toward the top, ranging from 0.3 to 1.5 mm in length. Like O. elongatum , the flowering parts are arranged mostly in terminal spikes. Leaves are shortly petiolate at the base, becoming subsessile toward the apex, arranged in up to 25 pairs per stem. Floral characteristics include staminal filaments up to 4 mm and styles reaching 8 mm. O. grosii flowers mainly between June and July (Bakha et al. 2019). Sampling and DNA Extraction Overall, 210 individual plants of Origanum elongatum and 54 individuals of O. grosii were sampled from geographically distinct locations spanning the full known distribution range of each species in Morocco. Species identification was confirmed using the taxonomic keys provided by Ietswaart (1980). Specimen collection in the field was carried out during flowering season (June-July). The sampling design aimed to capture maximum geographic and ecological variation. In most populations of O. elongatum , at least 25 individuals were sampled. However, in Boulmane region, only eight individuals could be located due to the low population density. Due to its extremely limited distribution, O. grosii is confined to a small area, which restricted the sampling to a single available population. O. elongatum and O. grosii samples were collected at distances ranging from 20 m to a few kilometers apart to minimize the risk of sampling clonal individuals. The sampled populations of O. elongatum cover its regional distribution across environmentally heterogeneous areas separated by major geographic barriers (deep valleys, Rif and Middle Atlas mountains) that may limit gene flow. Young leaves were collected and preserved at −80 °C, and genomic DNA was extracted from lyophilized tissue using the NucleoSpin® 96 Plant Kit following Aboukhalid et al. (2017). DNA quality was checked by 2.0% agarose gel electrophoresis, and purified DNA was stored at −20 °C for subsequent analyses. SSR Amplification and Genotyping Thirty SSR markers described by Ince et al. (2014) were screened on 56 individuals of O. elongatum and O. grosii , leading to the selection of 13 polymorphic markers with reliable amplification for genotyping. SSR genotyping employed M13-tailed primers following Schuelke (2000). Fluorescent labeling of PCR products was achieved by using M13 primers tagged with one of the four dyes: 6-FAM, VIC, PET, or NED (Eurofins Genomics, France). PCR amplification and genotyping procedures were conducted according to the protocol described by Aboukhalid et al. (2017). The resulting PCR products were analyzed with an ABI PRISM 3130 Genetic Analyzer, and GeneMapper software (Applied Biosystems) was used to analyze and determine fragments lengths. Genetic diversity Parameters including allele number (Na), observed (H O ) and expected heterozygosity (He), proportion of polymorphic loci (P%), Inbreeding coefficient (Fis) and genetic differentiation (Fst) were computed with GenAlEx 6.5 (Peakall and Smouse 2006). Allelic richness (Rs) and gene diversity (Hs) corrected for sample size (El Mousadik and Petit 1996) were calculated using FSTAT v. 2.9.3.2 (Goudet, 1995). The Hardy-Weinberg Equilibrium (HWE) analysis of the data was conducted using the Genepop v.1.2 (Raymond and Rousset 1995; Rousset 2008). Bottleneck effects were assessed using Bottleneck v.1.2.02 under infinite alleles model (IAM), stepwise mutation model (SMM), and two-phase model (TPM), applying Wilcoxon tests for heterozygosity excess and mode-shift analyses of allele frequency distributions. Genetic Structure The population genetic structure of O. elongatum and O. grosii was analyzed following the genotyping framework reported by Aboukhalid et al. (2017). Genetic variation within and among populations was assessed using Principal Coordinate Analysis (PCoA) using GenAlEx v6.4.1 (Peakall and Smouse 2006), while population structure was inferred through Bayesian clustering in STRUCTURE v2.3.4 (Pritchard et al. 2000). Model selection and visualization were performed using STRUCTURE HARVESTER (Evanno et al. 2005), CLUMPP (Jakobsson and Rosenberg 2007) and DISTRUCT (Rosenberg, 2003). Results Genetic Diversity In total, 264 O. elongatum and O. grosii samples were examined using 13 highly polymorphic microsatellite markers. Out of these markers, a total of 92 alleles were identified, with an average of 7.1 alleles per locus. The average observed heterozygosity was HO = 0.439, slightly higher than the expected heterozygosity He = 0.415 for the entire set of samples (Table 1). This shows the high genetic diversity within the samples from the two Origanum species. However, four markers (LB004, LB035, LB039, and LB077) showed significant deviations from the Hardy-Weinberg equilibrium (p < 0.05) and high positive Fis values (Table 1). This could be due to heterozygote deficiencies, which could be the result of inbreeding, the presence of null alleles, or the Wahlund effect, i.e., the substructuring of the population. Nevertheless, the high number of alleles and the heterozygosity indicate the high genetic diversity in the two Origanum species, which could be the result of the ecological adaptability of these plants and potential for gene flow across populations. Table 1 Summary of genetic diversity parameters estimated from 13 SSR loci in 264 individuals sampled from seven populations of O. elongatum and O. grosii . Loci Na Ne Ho He Fis Fst Nm HWE LB003 2.571 1.851 0.757 0.455 -0.664 0.049 4.681 ns LB004 5.286 3.866 0.556 0.739 0.248 0.062 3.450 * LB028 4.714 1.832 0.511 0.415 -0.230 0.156 1.535 ns LB035 8.857 2.568 0.420 0.539 0.222 0.086 1.740 * LB039 3.571 1.310 0.144 0.200 0.279 0.105 2.373 * LB040 4.714 1.920 0.550 0.445 -0.236 0.052 4.364 ns LB043 3.429 2.465 0.708 0.577 -0.228 0.218 1.230 ns LB060 3.429 1.205 0.164 0.159 -0.034 0.053 4.278 ns LB066 2.143 1.417 0.286 0.261 -0.097 0.284 0.560 ns LB070 1.714 1.073 0.069 0.064 -0.071 0.036 6.634 ns LB077 3.000 1.818 0.234 0.406 0.423 0.278 0.736 * LB078 3.429 2.553 0.511 0.571 0.105 0.099 1.616 ns LB081 3.000 2.057 0.687 0.494 -0.392 0.052 4.615 ns Mean 1.863 2.022 0.439 0.415 -0.053 0.119 2.909 Na : mean number of alleles, Ne : effective allele number, Ho : observed heterozygosity, He : expected heterozygosity, and Fis : inbreeding coefficient, computed as 1 minus Ho divided by He. HWE : the overall test for Hardy–Weinberg equilibrium deviations, with an asterisk (*) marking significant departures at the 0.05 level, and “ns” indicating non-significance. The genetic diversity of six O. elongatum populations and one O. grosii population was assessed (Table 2). At the population scale, allele numbers (Na) per locus ranged between 2.538 in O.e 1 (Boulmane region) and 4.692 in O.e 5 (Taounate region). On average, the estimated effective number of alleles (Ne) reached 1.995. Genetic diversity varied substantially among populations. Observed heterozygosity (Ho) ranged from 0.285 in O. grosii to 0.505 in population O.e 5, while expected heterozygosity (He) varied from 0.291 ( O. grosii ) to 0.495 ( O.e 5). The mean number of alleles per locus (Na) across 13 loci ranged between 2.538 ( O.e 1) and 4.692 ( O.e 5), and allelic richness (Rs) from 2.228 ( O. grosii ) to 2.989 ( O.e 5). Gene diversity (Hs) spanned 0.294 ( O. grosii ) to 0.500 ( O.e 5). A total of 26 private alleles were detected in 6 of 7 populations, with O. grosii containing the highest number (10). Overall, 94.51% of loci were polymorphic, with the lowest polymorphism in O.e 1 (76.92%) and complete polymorphism (100%) in populations O.e3 , O.e 4, O.e 5, and O.e 6. Table 2 Genetic diversity measures for populations of O. elongatum and O. grosii . Species Population code Region Sample size Na Rs Hs Np (P)% Ne Ho He Fis HWE O. elongatum O.e 1 Boulmane 8 2.538 2.524 0.427 - 76.92 2.035 0.470 0.402 LB004*** O. elongatum O.e 2 Chefchaouen 25 3.154 2.468 0.410 1 92.31 1.879 0.462 0.403 LB077** O. elongatum O.e 3 Targuist 65 4.615 2.836 0.399 6 100 2.149 0.374 0.395 *** LB004*** LB035*** LB039** O. elongatum O.e 4 Al Hoceima 36 4.231 2.864 0.444 3 100 2.088 0.479 0.437 * LB004** LB035*** LB077* O. elongatum O.e 5 Taounate 44 4.692 2.989 0.500 5 100 2.182 0.505 0.495 ** LB004*** LB039*** LB077* O. elongatum O.e 6 Taza 32 3.846 2.814 0.451 1 100 2.061 0.439 0.443 *** LB004*** LB035*** LB039*** LB077** Mean 3.846 2.749 0.439 3.2 94.872 2.066 0.455 0.429 O. grosii O.g Chefchaouen 54 3.769 2.228 0.294 10 92.31 1.572 0.285 0.291 *** LB003** LB035*** LB039** N : number of individuals analyzed. Na : average allele per locus across the 13 SSR markers. Rs : allelic richness calculated following El Mousadik and Petit (1996) [24]. Hs : mean gene diversity over all loci. Np : number of private alleles identified. P : proportion of loci that are polymorphic. Ho and He : observed and expected heterozygosity, respectively. FIS : inbreeding coefficient, with significance levels for heterozygote deficits indicated by * (P < 0.05), ** (P < 0.01), and *** (P < 0.001) at the population scale. HWE : loci that significantly deviate from Hardy–Weinberg equilibrium according to a global test at the 5% threshold, applying a sequential Bonferroni correction; stars mark loci with significant heterozygote deficiencies at the indicated significance levels. Population genetic structure based on microsatellite markers Principal Coordinate Analysis (PCoA) revealed clear genetic differentiation between O. elongatum and O. grosii (Fig. 1), with each species forming distinct clusters and showing substantial interspecific divergence. While a few individuals displayed partial overlap, likely due to shared alleles or limited gene flow, most samples clustered according to species and geographic origin, indicating species-specific gene pools and restricted genetic exchange. The pronounced separation along PCoA axes reflects evolutionary and ecological processes, potentially driven by geographic isolation, ecological specialization, or reproductive barriers. These results confirm that O. elongatum and O. grosii are genetically distinct, supporting their taxonomic status and underscoring the need to conserve their unique genetic resources. Bayesian Clustering and Genetic Structure STRUCTURE analysis identified four genetic clusters (ΔK; Evanno et al., 2005), with O. grosii forming a distinct cluster separate from O. elongatum . Within O. elongatum , populations O.e 2, O.e 4, O.e 5, and O.e 6 shared a genetic background, indicating a common gene pool or ongoing gene flow among them. In contrast, the most geographically isolated populations, O.e 1 and O.e 3, exhibited unique genetic compositions that clearly distinguished them from the other O. elongatum populations, although a few individuals showed signatures of admixture (Fig. 2b; Fig. 3). O. grosii , a species with a limited distribution, was genetically uniform and structured into a single distinct cluster. It is found in a fragmented habitat and appears clearly differentiated from the broader genetic pool of O. elongatum (Fig. 3). Pairwise genetic differentiation analysis corroborated the results and the distinctness of O. grosii with regard to O. elongatum . All the pairwise comparisons are significant even after correcting the results using the Bonferroni correction (p < 0.002). The genetic distances between the populations using Nei’s genetic distance ranged from 0.017 to 0.234 (Table 3). The maximum genetic distance was observed between O. grosii and O. elongatum from Taza (O.e6), while O. elongatum populations from Al Hoceima ( O.e 4) and Taza ( O.e 6) showed the lowest genetic distance. The genetic separation between O. elongatum and O. grosii and the effect of geographic continuity on the genetic similarity between O. elongatum populations are confirmed. The strong differentiation of O. grosii likely results from historical isolation, restricted gene flow, and species-specific evolutionary history. This strong genetic divergence supports its status as a distinct taxon and emphasizes the need for species-specific conservation strategies. Table 3 Pairwise comparison matrix of Nei genetic distance for O. elongatum and O. grosii populations. Populations O. e1 O. e2 O. e3 O. e4 O. e5 O. e6 O. g O.e1 **** O.e2 0.178 **** O.e3 0.096 0.197 **** O.e4 0.100 0.036 0.114 **** O.e5 0.108 0.055 0.141 0.036 **** O.e6 0.099 0.052 0.096 0.017 0.024 **** O.g 0.174 0.225 0.128 0.221 0.208 0.234 **** Mutation-Drift Equilibrium The genetic signatures of the recent decline in the effective population size are usually characterized by reduced allelic richness and heterozygosity, with allelic richness being reduced at a faster rate, which could result in observed heterozygosity being higher than the mutation drift equilibrium (Cornuet and Luikart 1996). Bottleneck analysis was carried out using the IAM, SMM, and TPM models with Wilcoxon’s signed-rank test for heterozygosity excess and the mode-shift test to identify changes in allele frequency distribution. Among the populations surveyed, O.e 1 showed a change in allele frequency distribution and heterozygosity excess (p < 0.05) using all three models (Table 4). The bottleneck could be due to the reduced size of the effective population. Other populations showed the characteristic L-shaped curve, which is the expected curve in the absence of a bottleneck event. However, care must be taken while interpreting the results, and the presence of outbreeding depression, Hardy-Weinberg equilibrium, null alleles, and inbreeding could affect the results (Aboukhalid et al. 2017). Table 4 Summary of the Wilcoxon signed-rank test outcomes evaluating excess heterozygosity and allele frequency distribution shifts under the infinite alleles model (IAM), the stepwise mutation model (SMM), and the two-phase model (TPM). Significant deviations from mutation-drift equilibrium are indicated by p -values less than 0.05. Populations IAM model TPM model SMM model Mode shift O. e1 0.00049 0.00244 0.01221 Shifted mode O.e2 0.16968 0.45483 0.66138 Normal L- shaped O.e3 0.58032 0.90454 0.99573 Normal L- shaped O.e4 0.47302 0.72913 0.90454 normal L- shaped O.e5 0.29395 0.55371 0.79285 normal L- shaped O.e6 0.29395 0.70605 0.96594 normal L- shaped O.g 0.84937 0.95386 0.99329 normal L- shaped Discussion Patterns of Genetic Diversity Small or narrowly distributed plant species are often susceptible to declines in genetic diversity, making their study essential for conservation planning. In this study, we assessed the genetic diversity of two Moroccan endemics, O. elongatum and O. grosii , using 13 microsatellite markers. Observed heterozygosity (Ho) ranged from 0.285 in O. grosii to 0.505 in O. elongatum ( O.e 5). Expected heterozygosity (He) was lower than Ho in four populations ( O.e 1, O.e 2, O.e 4, O.e 5), suggesting heterozygote excess. On average, O. elongatum exhibited higher genetic diversity (Ho = 0.455; He = 0.429) than O. compactum from Morocco (Ho = 0.37; He = 0.35; Aboukhalid et al. 2017) and comparable values to O. vulgare subsp. hirtum from Bulgaria (He = 0.482; Ho = 0.448; Alekseeva et al. 2021). In comparison with other species of the Origanum genus , O. elongatum showed exceptionally high levels of genetic diversity. In O. compactum , where the same SSR markers were used, the number of alleles per locus was 2.26, allelic richness was 1.72, and there were 14 private alleles (Aboukhalid et al. 2017). This study supported that low levels of genetic diversity are not universal among range-restricted species. Some range-restricted species may harbor levels of diversity comparable to or even higher than those of their common congeners (Gitzendanner and Soltis 2000). The high levels of genetic diversity found in O. elongatum , despite its range-restricted nature, may indicate ecological and genetic resilience that are likely linked to broad substrate adaptability and lack of recent founder events. O. grosii showed lower levels of genetic diversity, especially regarding allelic richness compared to O. elongatum . This pattern is comparable to that found for other range-restricted endemic species (Gibson et al. 2008). Confined to a single locality , O. grosii illustrates this trend. Similar patterns of decreased genetic diversity have been recorded for other range-restricted species such as Seseli farrenyi (Garcia-Jacas et al. 2021). Although O. grosii is a perennial species, and thus not subject to bottlenecks due to short life cycles, its small effective population size increases susceptibility to genetic drift, leading to greater homozygosity (Gustafsson and Sjögren-Gulve 2002). Surveys revealed no additional populations of O. grosii , confirming its critical status and the need to prioritize its single population for conservation. Its heterozygote deficit may partly reflect null alleles, often overestimated in small or inbred populations (Van Oosterhout et al. 2005). Reduced genetic diversity in O. grosii and O. elongatum relative to widespread O. vulgare is consistent with effects of drift, inbreeding, and allelic loss in small populations (Spielman et al. 2004; Alekseeva et al. 2021). However, genetic diversity also depends on life history, breeding system, gene flow, dispersal, and historical biogeography (Wang et al. 2004). For O. grosii , low diversity is not explained by its perennial, allogamous nature; pollinator limitation or ecological specialization may contribute (Phillips et al. 2014). In O. elongatum , heterozygote deficits in O.e2 and O.e6 may reflect mating among relatives. Genetic differentiation between O. elongatum and O. grosii is supported by species-specific alleles, with population assignment matching geographic origin even in sympatric areas like Chefchaouen. Admixed genotypes indicate past or ongoing gene flow, reflecting divergence with occasional interspecific exchange. The strong heterozygote deficit in O.e1 may result from null alleles (Arbeláez-Cortés et al. 2007), inbreeding or drift (Dodd et al. 2002), or a Wahlund effect from population structure (Dharmarajan et al. 2012). Population Structure Analysis The results of the Bayesian clustering analysis revealed the presence of four genetic clusters, (K = 4). The results of the STRUCTURE analysis revealed the presence of three lineages of O. elongatum and one lineage of O. grosii , with admixed populations showing evidence of historical or recent gene flow. The results were further confirmed by the Principal Coordinate Analysis (PCoA), which aligned with the inferred genetic structure and population assignments. In O. elongatum , the populations O.e 2, O.e 4, O.e 5, and O.e 6 were grouped together, showing evidence of evolutionary history or recurrent gene flow, which might be attributed to historical or recent gene flow, founder effects, and expansion events. In accordance with our findings, a previous study carried out by SSR markers on O. compactum from Morocco, revealed that founder effects and bottlenecks were responsible for the lower levels of genetic diversity (Aboukhalid et al. 2017). Bottleneck analysis detected evidence of a recent demographic decline only in population O.e 1, indicated by a mode-shifted allele frequency distribution and heterozygosity excess, although the small sample size (n = 8) may limit the robustness of this inference (Cornuet and Luikart 1996). The decline nevertheless appears plausible given the observed habitat degradation in this area. Genetic structure mirrors ecological boundaries, likely driven by restricted gene flow (Epps and Keyghobadi, 2015). Admixture indicates past or ongoing gene flow, and O. grosii remains sufficiently related to O. elongatum for cross-species SSR amplification. Increasing habitat fragmentation from land-use change threatens gene flow and may accelerate genetic erosion in narrow endemics, highlighting the need to combine land-cover mapping and genetics to prioritize connectivity and conservation (Epps and Keyghobadi, 2015). Conservation Implications This study provides the first comprehensive genetic baseline for O. grosii and O. elongatum , demonstrating that conserving genetic diversity is essential for long-term persistence and identifying priority populations beyond morphological and geographic criteria. Genetically diverse and distinct populations (notably O.e 4 and O.e 5) should be prioritized for in situ conservation and integrated into ex situ programs, while the widespread presence of private alleles (except O.e 1) enhances most populations’ value for restoration and reintroduction (Frankham 2016). Four genetic clusters were identified, supporting their management as separate Evolutionarily Significant Units (ESUs) and Management Units (MUs). While populations within clusters (e.g., O.e 2, O.e 4, O.e 5, O.e 6) may serve as mutual reinforcement sources, inter-cluster mixing should be avoided to reduce the risk of outbreeding depression (Waller 2015; Moritz 1994). Urgent genetic reinforcement and habitat restoration are needed for the genetically impoverished Boulemane population (O.e1). Ex situ conservation and use of local seed sources are advised to preserve local adaptation (Quilichini et al. 2004). Major threats including overgrazing, agriculture, urbanization, and illegal harvesting highlight the need for legal protection, inclusion in Key Biodiversity Areas, and alignment with IUCN assessments. Although fragmented, O. elongatum maintains relatively high genetic diversity but limited gene flow, calling for targeted management. Integrating population genetics with ecological and climate modeling is essential to conserve adaptive capacity and ensure long-term viability under climate change (Hoffmann and Sgrò 2021), providing a transferable framework for conserving other narrowly distributed endemic species. Conclusion This study provides the first comprehensive genetic assessment of O. elongatum and O. grosii across Morocco, revealing clear inter- and intraspecific differentiation with key conservation implications. Despite habitat fragmentation, O. elongatum maintains relatively high genetic diversity, though limited gene flow among populations calls for targeted actions. Populations in Taounate and Al Hoceima, with high diversity and unique genetic profiles, should be prioritized, while isolated, genetically poor populations like Boulemane need urgent reinforcement and habitat restoration. Four genetic clusters support their designation as separate management units for restoration programs. The findings underscore the value of integrating molecular data with conservation planning, considering geographic isolation, ecological gradients, and land-use pressures, to guide strategies that preserve evolutionary potential and adaptive resilience in Morocco’s endemic flora and other threatened species in fragile regions. Declarations Acknowledgments We express our sincere gratitude to the “Service de Systématique Moléculaire” (UMS 2700, Muséum National d'Histoire Naturelle, Paris, France) for providing access to their laboratories for the molecular genetic analyses. We also gratefully acknowledge the financial support provided by the Moroccan-French cooperation program (Toubkal). Author contribution K.A., N.M., A.L. and C.A.F. designed and conceptualized the study. K.A. performed formal analyses and wrote the original draft. K.A., M.B. and A.K. conducted fieldwork, sample collection and identified plant material. J.L. conceived the laboratory analyses and participated in DNA extraction, PCR amplification, and SSR genotyping. M.N., A.M. and G.K.G. performed statistical analyses, prepared some figures and contributed to manuscript review and editing. N.M., C.A.F. and A.L. supervised the project. N.M. and C.A.F. acquired funding and administered the project. All authors gave final approval for publication. Funding This work was partially supported by the “PHC Toubkal” programme, funded by the French Ministry for Europe and Foreign Affairs, the French Ministry for Higher Education Research and Space (MESRE), and the Moroccan Ministry for Higher Education,Scientific Research and Innovation (MESRSI). Data Availability The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request. Conflict of interest The authors confirm that there are no financial conflicts or personal connections that might have influenced the research presented in this article. References Aboukhalid K, Al Faiz C, Bakha M, Douaik A, Lamiri A, Gibernau M, Tomi F (2023) Agronomic evaluation and essential oil composition characterization of Origanum compactum accessions cultivated in Morocco. 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Mol Ecol Resour 8:103–106. https://doi.org/10.1111/j.1471-8286.2007.01931.x Simon JE, Alena FC, Lyle EC (1984) Herbs: an indexed bibliography 1971–1980, the scientific literature on selected herbs, and aromatic and medicinal plants of the temperate zone . Archon Books, HSA Library Spielman D, Brook BW, Briscoe DA, Frankham R (2004) Does Inbreeding and Loss of Genetic Diversity Decrease Disease Resistance? Conserv Genet 5:439–448. https://doi.org/10.1023/b:coge.0000041030.76598.cd Sullivan ER, Barker C, Powell I, Ashton PA (2019) Genetic diversity and connectivity in fragmented populations of Rhinanthus minor in two regions with contrasting land-use. Biodivers Conserv 28:3159–3181. https://doi.org/10.1007/s10531-019-01811-x Supple MA, Shapiro B (2018) Conservation of biodiversity in the genomics era. Genome Biol 19. https://doi.org/10.1186/s13059-018-1520-3 Van Oosterhout C, Weetman D, Hutchinson WF (2005) Estimation and adjustment of microsatellite null alleles in nonequilibrium populations. Mol Ecol Notes 6:255–256. https://doi.org/10.1111/j.1471-8286.2005.01082.x Waller DM (2015) Genetic rescue: a safe or risky bet? Mol Ecol 24:2595–2597. https://doi.org/10.1111/mec.13220 Wang Z-F (2004) High Genetic Diversity in Sarracenia leucophylla (Sarraceniaceae), a Carnivorous Wetland Herb. J Hered 95:234–243. https://doi.org/10.1093/jhered/esh043 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8870849","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":598296022,"identity":"0219fad6-417f-4dee-ab10-ea945bb07333","order_by":0,"name":"Kaoutar Aboukhalid","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABD0lEQVRIie3PMUsDMRjG8VcKcXmxa0Il/QTCuQRd+lkCgZviIC5ChSqF3KSzxS/Rbg4OOQLpUrn1xCIVoZOCIjiKKSJ0iFo3h/yn444fz3sAqdQ/jC4/rZ0cwyYBZe2vxC4RJJDLPxIAnf14GCucbz0f3UHzou/Z4LKDG+v6xe1fdWCriI+1MM+p9QdApz5no4lCgk9Ddz5XICYySjhoQS0JH2st2L1pIKF7Q4e2AcJ+Q5qPgbxLaH+SXiB6FkgPRDWLH0bDSmkkZAsyMm5BIBAHoo6vsHD1zvWZxO2pV7sDMw7/Ms8CGaOo4yu0UmV9+CY5v+2XN6emy9uFenhF2+Wiiq98hSu8SaVSqdTKfQA+zGD+5kK+nQAAAABJRU5ErkJggg==","orcid":"","institution":"Institut National de la Recherche Agronomique","correspondingAuthor":true,"prefix":"","firstName":"Kaoutar","middleName":"","lastName":"Aboukhalid","suffix":""},{"id":598296023,"identity":"27f8adf0-e861-409b-8032-173ac3834e60","order_by":1,"name":"Nathalie Machon","email":"","orcid":"","institution":"Muséum National d'Histoire Naturelle","correspondingAuthor":false,"prefix":"","firstName":"Nathalie","middleName":"","lastName":"Machon","suffix":""},{"id":598296024,"identity":"22cb45ab-9cc6-4757-8b0b-ebcbcba86674","order_by":2,"name":"Josie Lambourdière","email":"","orcid":"","institution":"Muséum National d'Histoire Naturelle","correspondingAuthor":false,"prefix":"","firstName":"Josie","middleName":"","lastName":"Lambourdière","suffix":""},{"id":598296025,"identity":"3ad90d65-42ec-4a44-9f28-c79b4101bcca","order_by":3,"name":"Mohamed Bakha","email":"","orcid":"","institution":"University Sultan Moulay Slimane of Beni-Mellal","correspondingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"","lastName":"Bakha","suffix":""},{"id":598296027,"identity":"c699518b-e747-4e01-a8d4-34584ddb35c5","order_by":4,"name":"Mounsef Neffa","email":"","orcid":"","institution":"Mohamed I University","correspondingAuthor":false,"prefix":"","firstName":"Mounsef","middleName":"","lastName":"Neffa","suffix":""},{"id":598296028,"identity":"5bfdc119-e334-4bf3-a36a-e2b494d2ca6d","order_by":5,"name":"Grazyna Korbecka-Glinka","email":"","orcid":"","institution":"Institute of Soil Science and Plant Cultivation","correspondingAuthor":false,"prefix":"","firstName":"Grazyna","middleName":"","lastName":"Korbecka-Glinka","suffix":""},{"id":598296029,"identity":"e80f3f06-9a85-4f94-910e-a49fc7b27f50","order_by":6,"name":"Abdelkarim Khiraoui","email":"","orcid":"","institution":"University Sultan Moulay Slimane of Beni-Mellal","correspondingAuthor":false,"prefix":"","firstName":"Abdelkarim","middleName":"","lastName":"Khiraoui","suffix":""},{"id":598296030,"identity":"2b47b55b-fcc8-463d-b527-4fcfa8083067","order_by":7,"name":"Ravish Choudhary","email":"","orcid":"","institution":"Indian Agricultural Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Ravish","middleName":"","lastName":"Choudhary","suffix":""},{"id":598296031,"identity":"ae22f53d-42ec-40cf-89d2-ef86d1b6889d","order_by":8,"name":"Abdeslam Lamiri","email":"","orcid":"","institution":"Université Hassan 1er","correspondingAuthor":false,"prefix":"","firstName":"Abdeslam","middleName":"","lastName":"Lamiri","suffix":""},{"id":598296032,"identity":"06b29b16-9362-4ee5-9e31-b9b8e44e8c2a","order_by":9,"name":"Chaouki Al Faiz","email":"","orcid":"","institution":"Institut National de la Recherche Agronomique","correspondingAuthor":false,"prefix":"","firstName":"Chaouki","middleName":"Al","lastName":"Faiz","suffix":""}],"badges":[],"createdAt":"2026-02-13 10:53:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8870849/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8870849/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103894990,"identity":"ac9a9251-da27-4e64-820c-19cf04cbf6ae","added_by":"auto","created_at":"2026-03-04 08:46:07","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":127699,"visible":true,"origin":"","legend":"\u003cp\u003ePrincipal Coordinates Analysis (PCoA) illustrating genetic relationships among individuals. The first and second axes account for 15.92% and 12.13% of the total genetic variation, respectively.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8870849/v1/8ca00f4594eb34552eecc633.png"},{"id":103894992,"identity":"0d98b5a7-b8ab-4ce5-a03d-1332d416df8e","added_by":"auto","created_at":"2026-03-04 08:46:07","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":80270,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea \u003c/strong\u003eBayesian clustering results used to estimate the most likely number of genetic groups, following the method of Evanno et al. (2005) [29]. The ΔK value, representing the second-order rate of change in log-likelihood between successive K values, was calculated to identify the optimal K. Both ln(K) and ΔK were analyzed using STRUCTURE HARVESTER [32] to determine the best-supported number of clusters.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eb\u003c/strong\u003e Bayesian clustering analysis performed using the STRUCTURE software [28]. The bar plots illustrate the inferred genetic structure of \u003cem\u003eO. elongatum \u003c/em\u003eand \u003cem\u003eO. grosii\u003c/em\u003e, assuming four clusters (K = 4). Each bar on the Y-axis corresponds to an individual, with its genetic ancestry depicted by color-coded segments indicating proportional membership in the inferred clusters. Thin vertical lines separate individuals within populations, while bold lines distinguish different populations.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8870849/v1/519dd39965e4c492562d74e9.png"},{"id":103894991,"identity":"5af3f59d-96d4-4992-bcca-cbae55385559","added_by":"auto","created_at":"2026-03-04 08:46:07","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":254899,"visible":true,"origin":"","legend":"\u003cp\u003eSpatial distribution and inferred genetic structure of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e across seven populations. Pie charts indicate the average cluster membership proportions for each population, based on the Bayesian clustering results at K = 4, as determined by STRUCTURE (corresponding to Fig. 2b).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8870849/v1/1d6b8ff90bf1b826b73d2cd9.png"},{"id":104401686,"identity":"925d172c-15e3-4031-b474-cd169133c33a","added_by":"auto","created_at":"2026-03-11 12:13:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1479443,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8870849/v1/26050035-e73f-44d9-8fbe-d3bb694e7635.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Genetic diversity and differentiation of two narrowly endemic Origanum Species in Morocco: implications for conservation","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMorocco is a major biodiversity hotspot in the Mediterranean, hosting about 3,912 taxa and 1,298 subspecies, with roughly 22% endemism (Fennane and Ibn Tattou \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This exceptional endemic richness (878 taxa, nearly 600 at the species level) is mainly concentrated in the High Atlas, Middle Atlas, and Rif mountains. However, this diversity is increasingly threatened, as nearly 2,193 plant species are endangered, over half of which (53%) are highly vulnerable due to anthropogenic pressures and environmental change (Fennane and Ibn Tattou \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). Endemics with narrow distributions are particularly at risk, as their limited range increases their vulnerability to stochastic events and environmental stressors (George et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Lannuzel et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe genus \u003cem\u003eOriganum\u003c/em\u003e L. (Lamiaceae), comprising about 18 hybrids and 43 species, is largely distributed around the Mediterranean region (Ietswaart, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). In Morocco, four taxa and one hybrid are recognized: \u003cem\u003eO. compactum\u003c/em\u003e Benth. (also found in southern Spain), \u003cem\u003eO. vulgare\u003c/em\u003e subsp. \u003cem\u003evirens\u003c/em\u003e, and three endemic taxa : \u003cem\u003eO. elongatum\u003c/em\u003e, \u003cem\u003eO. grosii\u003c/em\u003e, and \u003cem\u003eO. \u0026times; font-queri\u003c/em\u003e (a spontaneously occurring hybrid resulting from \u003cem\u003eO. grosii\u003c/em\u003e and \u003cem\u003eO. compactum\u003c/em\u003e) (Bakha et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The species \u003cem\u003eO. elongatum\u003c/em\u003e has a geographic range that stretches mainly across northeastern Morocco, between the Middle Atlas and Rif Mountain systems, typically at high elevations (Bakha et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003eO. grosii\u003c/em\u003e is among the rarest species, occurring in small, isolated populations in the humid western Rif at high altitudes (Bakha et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTaxonomic delimitation within Moroccan \u003cem\u003eOriganum\u003c/em\u003e is controversial. Morphological similarities among \u003cem\u003eO. grosii\u003c/em\u003e, \u003cem\u003eO. elongatum\u003c/em\u003e and the hybrid \u003cem\u003eO. \u0026times; font-queri\u003c/em\u003e have led to confusion. Although certain databases (Euro\u0026thinsp;+\u0026thinsp;Med \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; African Plant Database v3.4.0, Dobignard and Chatelain 2010\u0026ndash;2013) classify \u003cem\u003eO. grosii\u003c/em\u003e and the hybrid \u003cem\u003eO. \u0026times; font-queri\u003c/em\u003e under the same taxon as \u003cem\u003eO. elongatum\u003c/em\u003e (Bakha et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), Ietswaart (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1980\u003c/span\u003e) designated \u003cem\u003eO. grosii\u003c/em\u003e as an independent species, based on morphological traits, including its shorter stems and leaves, as well as broader bracts in comparison to \u003cem\u003eO. elongatum\u003c/em\u003e.\u003c/p\u003e \u003cp\u003ePopulations of Moroccan \u003cem\u003eOriganum\u003c/em\u003e species are highly fragmented and increasingly threatened by drought, climate change, habitat loss, overgrazing, and deforestation (Aboukhalid et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Such fragmentation and population decline often result in enhanced genetic drift, reduced gene flow, and decline in genetic variability, all of which compromise the adaptive potential and long-term persistence of species (Holsinger and Gottlieb \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1991\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLow levels of genetic variability and high levels of genetic divergence among populations are common characteristics of rare and geographically isolated species (Sullivan et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Endemic medicinal plants like \u003cem\u003eOriganum\u003c/em\u003e species are highly vulnerable to habitat degradation and climate change, emphasizing the need for conservation (Aboukhalid et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Genetic diversity is crucial for adaptive capacity and long-term survival (Reed and Frankham \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Supple and Shapiro \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), yet small populations often lose diversity through drift, bottlenecks, and founder effects (Banks et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), making its conservation a central goal (Frankham \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). This study addresses the previously unexamined genetic diversity and population structure of Moroccan endemic \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e using genomic and landscape approaches across their natural range. In particular, we investigate the following key questions: (i) What are the patterns of genetic polymorphism and population structure in \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e across and within populations? (ii) Is there evidence of gene flow among these species? (iii) Do patterns of genetic differentiation support the current taxonomic distinctions proposed by Ietswaart (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1980\u003c/span\u003e)? (iv) How might the genetic results contribute to the development of effective conservation and management plans for these rare and endangered endemics?\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy Species\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eOriganum elongatum\u003c/em\u003e Emberger \u0026amp; Maire (Lamiaceae) is a perennial species native exclusively to Morocco. It is naturally distributed in Rif Mountain systems and the northern Middle Atlas, predominantly inhabiting schistose soils at elevations ranging from 681 to 1866 meters, with most populations occurring above 1300 m (Bakha et al. 2020). \u003cem\u003eO. elongatum\u003c/em\u003e is a climate-tolerant species found from semi-arid to per-humid zones and is valued in Morocco for its aromatic, culinary, melliferous, and ornamental uses. It is charachterized by tall purplish stems reaching up to 90 cm. Branches are largely composed of terminal spikes. Leaves are arranged in up to 30 opposite pairs per stem, becoming subsessile toward the apex, with petioles up to 5 mm in length (Ietswaart 1980). The flowering occurs from June to October. The leaves are densely encrusted with glandular trichomes that produce essential oils, resulting in a fragrant and flavored odor characteristically associated with this species (Simon et al. 1984).\u003c/p\u003e\n\u003cp\u003eAlthough specific pollination studies are lacking, flower morphology, strong scent, prominent stamens (up to 3.5 mm), and elongated styles (up to 8 mm), as well as the frequent visits by pollinating insects suggest a predominance of cross-pollination. This is consistent with the general reproductive patterns reported in the Lamiaceae family, which is typically insect-pollinated and predominantly outcrossing (Judd et al. 1999).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eOriganum grosii\u003c/em\u003e Pau \u0026amp; Font Quer ex Ietswaart stands out as one of the scarcest and most localized species taxonomically assigned to the genus \u003cem\u003eOriganum\u003c/em\u003e. This species is found only in the western Rif Mountains of northwestern Morocco. It grows mainly at high elevations under humid conditions, typically on calcareous substrates, at elevations ranging between 400 and 1900 m (Bakha et al. 2017).\u003c/p\u003e\n\u003cp\u003eMorphologically, \u003cem\u003eO. grosii\u003c/em\u003e is quite distinctive. It has relatively short stems (up to 55 cm), which are lightly hairy, and long lateral branches stretching between 4 and 30 cm. Its bracts are notably larger (2\u0026ndash;5 mm) than those of related species. The leaves are smaller than those of \u003cem\u003eO. elongatum\u003c/em\u003e, usually between 5 and 19 mm long. The species has erected purplish-brown stems, with fine hairs (trichomes) concentrated at the base and continuing sparsely toward the top, ranging from 0.3 to 1.5 mm in length. Like \u003cem\u003eO. elongatum\u003c/em\u003e, the flowering parts are arranged mostly in terminal spikes. Leaves are shortly petiolate at the base, becoming subsessile toward the apex, arranged in up to 25 pairs per stem. Floral characteristics include staminal filaments up to 4 mm and styles reaching 8 mm. \u003cem\u003eO. grosii\u003c/em\u003e flowers mainly between June and July (Bakha et al. 2019).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSampling and DNA Extraction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOverall, 210 individual plants of \u003cem\u003eOriganum elongatum\u003c/em\u003e and 54 individuals of \u003cem\u003eO. grosii\u003c/em\u003e were sampled from geographically distinct locations spanning the full known distribution range of each species in Morocco. Species identification was confirmed using the taxonomic keys provided by Ietswaart (1980). Specimen collection in the field was carried out during flowering season (June-July).\u003c/p\u003e\n\u003cp\u003eThe sampling design aimed to capture maximum geographic and ecological variation. In most populations of \u003cem\u003eO. elongatum\u003c/em\u003e, at least 25 individuals were sampled. However, in Boulmane region, only eight individuals could be located due to the low population density. Due to its extremely limited distribution, \u003cem\u003eO. grosii\u003c/em\u003e is confined to a small area, which restricted the sampling to a single available population. \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e samples were collected at distances ranging from 20 m to a few kilometers apart to minimize the risk of sampling clonal individuals. The sampled populations of \u003cem\u003eO. elongatum\u003c/em\u003e cover its regional distribution across environmentally heterogeneous areas separated by major geographic barriers (deep valleys, Rif and Middle Atlas mountains) that may limit gene flow. Young leaves were collected and preserved at \u0026minus;80 \u0026deg;C, and genomic DNA was extracted from lyophilized tissue using the NucleoSpin\u0026reg; 96 Plant Kit following Aboukhalid et al. (2017). DNA quality was checked by 2.0% agarose gel electrophoresis, and purified DNA was stored at \u0026minus;20 \u0026deg;C for subsequent analyses.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSSR Amplification and Genotyping\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThirty SSR markers described by Ince et al. (2014) were screened on 56 individuals of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e, leading to the selection of 13 polymorphic markers with reliable amplification for genotyping. SSR genotyping employed M13-tailed primers following Schuelke (2000). Fluorescent labeling of PCR products was achieved by using M13 primers tagged with one of the four dyes: 6-FAM, VIC, PET, or NED (Eurofins Genomics, France). PCR amplification and genotyping procedures were conducted according to the protocol described by Aboukhalid et al. (2017). The resulting PCR products were analyzed with an ABI PRISM 3130 Genetic Analyzer, and GeneMapper software (Applied Biosystems) was used to analyze and determine fragments lengths.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenetic diversity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eParameters including allele number (Na), observed (H\u003csub\u003eO\u003c/sub\u003e) and expected heterozygosity (He), proportion of polymorphic loci (P%), Inbreeding coefficient (Fis) and genetic differentiation (Fst) were computed with GenAlEx 6.5 (Peakall and Smouse 2006). Allelic richness (Rs) and gene diversity (Hs) corrected for sample size (El Mousadik and Petit 1996) were calculated using FSTAT v. 2.9.3.2 (Goudet, 1995). The Hardy-Weinberg Equilibrium (HWE) analysis of the data was conducted using the Genepop v.1.2 (Raymond and Rousset 1995; Rousset 2008). Bottleneck effects were assessed using Bottleneck v.1.2.02 under infinite alleles model (IAM), stepwise mutation model (SMM), and two-phase model (TPM), applying Wilcoxon tests for heterozygosity excess and mode-shift analyses of allele frequency distributions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenetic Structure\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe population genetic structure of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e was analyzed following the genotyping framework reported by Aboukhalid et al. (2017). Genetic variation within and among populations was assessed using Principal Coordinate Analysis (PCoA) using GenAlEx v6.4.1 (Peakall and Smouse 2006), while population structure was inferred through Bayesian clustering in STRUCTURE v2.3.4 (Pritchard et al. 2000). Model selection and visualization were performed using STRUCTURE HARVESTER (Evanno et al. 2005), CLUMPP (Jakobsson and Rosenberg 2007) and DISTRUCT (Rosenberg, 2003).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eGenetic Diversity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn total, 264 \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e samples were examined using 13 highly polymorphic microsatellite markers. Out of these markers, a total of 92 alleles were identified, with an average of 7.1 alleles per locus. The average observed heterozygosity was HO = 0.439, slightly higher than the expected heterozygosity He = 0.415 for the entire set of samples (Table 1). This shows the high genetic diversity within the samples from the two \u003cem\u003eOriganum\u003c/em\u003e species. However, four markers (LB004, LB035, LB039, and LB077) showed significant deviations from the Hardy-Weinberg equilibrium (p \u0026lt; 0.05) and high positive Fis values (Table 1). This could be due to heterozygote deficiencies, which could be the result of inbreeding, the presence of null alleles, or the Wahlund effect, i.e., the substructuring of the population. Nevertheless, the high number of alleles and the heterozygosity indicate the high genetic diversity in the two \u003cem\u003eOriganum\u003c/em\u003e species, which could be the result of the ecological adaptability of these plants and potential for gene flow across populations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eSummary of genetic diversity parameters estimated from 13 SSR loci in 264 individuals sampled from seven populations of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLoci\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNa\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNe\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHo\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHe\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFis\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFst\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHWE\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB003\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.571\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.851\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.757\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.455\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.664\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.049\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e4.681\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e5.286\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.866\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.556\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.739\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.248\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.062\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e3.450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB028\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e4.714\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.832\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.511\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.415\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.230\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.156\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e1.535\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB035\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e8.857\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.568\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.420\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.539\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.086\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e1.740\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB039\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.571\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.310\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.144\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.279\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.105\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e2.373\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB040\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e4.714\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.920\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.550\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.445\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.236\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.052\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e4.364\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB043\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.429\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.465\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.708\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.228\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.218\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e1.230\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB060\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.429\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.205\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.159\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.034\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e4.278\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB066\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.143\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.417\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.286\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.261\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.097\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.284\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e0.560\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB070\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.714\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.069\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.064\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.071\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.036\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e6.634\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB077\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.818\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.234\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.406\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.423\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.278\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e0.736\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB078\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.429\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.553\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.511\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.571\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.105\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.099\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e1.616\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLB081\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e3.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.057\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.687\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.494\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.392\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.052\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e4.615\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003ens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e1.863\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e2.022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.439\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.415\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e-0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e0.119\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8153%;\"\u003e\n \u003cp\u003e2.909\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1481%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eNa\u003c/strong\u003e: mean number of alleles, \u003cstrong\u003eNe\u003c/strong\u003e: effective allele number, \u003cstrong\u003eHo\u003c/strong\u003e: observed heterozygosity, \u003cstrong\u003eHe\u003c/strong\u003e: expected heterozygosity, and \u003cstrong\u003eFis\u003c/strong\u003e: inbreeding coefficient, computed as 1 minus Ho divided by He. \u003cstrong\u003eHWE\u003c/strong\u003e: the overall test for Hardy\u0026ndash;Weinberg equilibrium deviations, with an asterisk (*) marking significant departures at the 0.05 level, and \u0026ldquo;ns\u0026rdquo; indicating non-significance.\u003c/p\u003e\n\u003cp\u003eThe genetic diversity of six \u003cem\u003eO. elongatum\u003c/em\u003e populations and one \u003cem\u003eO. grosii\u003c/em\u003e population was assessed (Table 2). At the population scale, allele numbers (Na) per locus ranged between 2.538 in \u003cem\u003eO.e\u003c/em\u003e1 (Boulmane region) and 4.692 in \u003cem\u003eO.e\u003c/em\u003e5 (Taounate region). On average, the estimated effective number of alleles (Ne) reached 1.995. Genetic diversity varied substantially among populations. Observed heterozygosity (Ho) ranged from 0.285 in \u003cem\u003eO. grosii\u003c/em\u003e to 0.505 in population \u003cem\u003eO.e\u003c/em\u003e5, while expected heterozygosity (He) varied from 0.291 (\u003cem\u003eO. grosii\u003c/em\u003e) to 0.495 (\u003cem\u003eO.e\u003c/em\u003e5). The mean number of alleles per locus (Na) across 13 loci ranged between 2.538 (\u003cem\u003eO.e\u003c/em\u003e1) and 4.692 (\u003cem\u003eO.e\u003c/em\u003e5), and allelic richness (Rs) from 2.228 (\u003cem\u003eO. grosii\u003c/em\u003e) to 2.989 (\u003cem\u003eO.e\u003c/em\u003e5). Gene diversity (Hs) spanned 0.294 (\u003cem\u003eO. grosii\u003c/em\u003e) to 0.500 (\u003cem\u003eO.e\u003c/em\u003e5). A total of 26 private alleles were detected in 6 of 7 populations, with \u003cem\u003eO. grosii\u003c/em\u003e containing the highest number (10). Overall, 94.51% of loci were polymorphic, with the lowest polymorphism in \u003cem\u003eO.e\u003c/em\u003e1 (76.92%) and complete polymorphism (100%) in populations \u003cem\u003eO.e3\u003c/em\u003e, \u003cem\u003eO.e\u003c/em\u003e4, \u003cem\u003eO.e\u003c/em\u003e5, and \u003cem\u003eO.e\u003c/em\u003e6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u0026nbsp;\u003c/strong\u003eGenetic diversity measures for populations of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"763\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulation code\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegion\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSample size\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNa\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNp\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e(P)%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNe\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHo\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHe\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFis\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHWE\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eBoulmane\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.538\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.524\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.427\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e76.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.035\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.470\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.402\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB004***\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eChefchaouen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e3.154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.468\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.410\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e92.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e1.879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.462\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.403\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB077**\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eTarguist\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e4.615\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.836\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.399\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.149\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.374\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.395\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB004***\u003c/p\u003e\n \u003cp\u003eLB035***\u003c/p\u003e\n \u003cp\u003eLB039**\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eAl Hoceima\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e4.231\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.864\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.444\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.088\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.479\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.437\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB004**\u003c/p\u003e\n \u003cp\u003eLB035***\u003c/p\u003e\n \u003cp\u003eLB077*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eTaounate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e4.692\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.182\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.505\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.495\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e**\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB004***\u003c/p\u003e\n \u003cp\u003eLB039***\u003c/p\u003e\n \u003cp\u003eLB077*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. elongatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.e\u003c/em\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eTaza\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e3.846\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.814\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.451\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.061\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.439\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB004***\u003c/p\u003e\n \u003cp\u003eLB035***\u003c/p\u003e\n \u003cp\u003eLB039***\u003c/p\u003e\n \u003cp\u003eLB077**\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\u0026nbsp;\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e3.846\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.749\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.439\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e3.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e94.872\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.066\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.455\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.429\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\u0026nbsp;\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11.8421%;\"\u003e\n \u003cp\u003e\u003cem\u003eO. grosii\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.1316%;\"\u003e\n \u003cp\u003e\u003cem\u003eO.g\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.1842%;\"\u003e\n \u003cp\u003eChefchaouen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 7.5%;\"\u003e\n \u003cp\u003e54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e3.769\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e2.228\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.294\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.07895%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 6.71053%;\"\u003e\n \u003cp\u003e92.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e1.572\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 5.78947%;\"\u003e\n \u003cp\u003e0.291\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4.47368%;\"\u003e\n \u003cp\u003e***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.34211%;\"\u003e\n \u003cp\u003eLB003**\u003c/p\u003e\n \u003cp\u003eLB035***\u003c/p\u003e\n \u003cp\u003eLB039**\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eN\u003c/strong\u003e: number of individuals analyzed. \u003cstrong\u003eNa\u003c/strong\u003e: average allele per locus across the 13 SSR markers. \u003cstrong\u003eRs\u003c/strong\u003e: allelic richness calculated following El Mousadik and Petit (1996) [24]. \u003cstrong\u003eHs\u003c/strong\u003e: mean gene diversity over all loci. \u003cstrong\u003eNp\u003c/strong\u003e:\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003enumber of private alleles identified. \u003cstrong\u003eP\u003c/strong\u003e: proportion of loci that are polymorphic. \u003cstrong\u003eHo\u003c/strong\u003e and \u003cstrong\u003eHe\u003c/strong\u003e: observed and expected heterozygosity, respectively. \u003cstrong\u003eFIS\u003c/strong\u003e: inbreeding coefficient, with significance levels for heterozygote deficits indicated by * (P \u0026lt; 0.05), ** (P \u0026lt; 0.01), and *** (P \u0026lt; 0.001) at the population scale. \u003cstrong\u003eHWE\u003c/strong\u003e: loci that significantly deviate from Hardy\u0026ndash;Weinberg equilibrium according to a global test at the 5% threshold, applying a sequential Bonferroni correction; stars mark loci with significant heterozygote deficiencies at the indicated significance levels.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePopulation genetic structure based on microsatellite markers\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePrincipal Coordinate Analysis (PCoA) revealed clear genetic differentiation between \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e (Fig. 1), with each species forming distinct clusters and showing substantial interspecific divergence. While a few individuals displayed partial overlap, likely due to shared alleles or limited gene flow, most samples clustered according to species and geographic origin, indicating species-specific gene pools and restricted genetic exchange. The pronounced separation along PCoA axes reflects evolutionary and ecological processes, potentially driven by geographic isolation, ecological specialization, or reproductive barriers. These results confirm that \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e are genetically distinct, supporting their taxonomic status and underscoring the need to conserve their unique genetic resources.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBayesian Clustering and Genetic Structure\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSTRUCTURE analysis identified four genetic clusters (\u0026Delta;K; Evanno et al., 2005), with \u003cem\u003eO. grosii\u003c/em\u003e forming a distinct cluster separate from \u003cem\u003eO. elongatum\u003c/em\u003e. Within \u003cem\u003eO. elongatum\u003c/em\u003e, populations \u003cem\u003eO.e\u003c/em\u003e2, \u003cem\u003eO.e\u003c/em\u003e4, \u003cem\u003eO.e\u003c/em\u003e5, and \u003cem\u003eO.e\u003c/em\u003e6 shared a genetic background, indicating a common gene pool or ongoing gene flow among them. In contrast, the most geographically isolated populations, \u003cem\u003eO.e\u003c/em\u003e1 and \u003cem\u003eO.e\u003c/em\u003e3, exhibited unique genetic compositions that clearly distinguished them from the other \u003cem\u003eO. elongatum\u003c/em\u003e populations, although a few individuals showed signatures of admixture (Fig. 2b; Fig. 3). \u003cem\u003eO. grosii\u003c/em\u003e, a species with a limited distribution, was genetically uniform and structured into a single distinct cluster. It is found in a fragmented habitat and appears clearly differentiated from the broader genetic pool of \u003cem\u003eO. elongatum\u003c/em\u003e (Fig. 3).\u003c/p\u003e\n\u003cp\u003ePairwise genetic differentiation analysis corroborated the results and the distinctness of \u003cem\u003eO. grosii\u003c/em\u003e with regard to \u003cem\u003eO. elongatum\u003c/em\u003e. All the pairwise comparisons are significant even after correcting the results using the Bonferroni correction (p \u0026lt; 0.002). The genetic distances between the populations using Nei\u0026rsquo;s genetic distance ranged from 0.017 to 0.234 (Table 3). The maximum genetic distance was observed between \u003cem\u003eO. grosii\u003c/em\u003e and \u003cem\u003eO. elongatum\u003c/em\u003e from Taza (O.e6), while \u003cem\u003eO. elongatum\u003c/em\u003e populations from Al Hoceima (\u003cem\u003eO.e\u003c/em\u003e4) and Taza (\u003cem\u003eO.e\u003c/em\u003e6) showed the lowest genetic distance. The genetic separation between \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e and the effect of geographic continuity on the genetic similarity between \u003cem\u003eO. elongatum\u003c/em\u003e populations are confirmed. The strong differentiation of \u003cem\u003eO. grosii\u003c/em\u003e likely results from historical isolation, restricted gene flow, and species-specific evolutionary history. This strong genetic divergence supports its status as a distinct taxon and emphasizes the need for species-specific conservation strategies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e Pairwise comparison matrix of Nei genetic distance for \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e populations.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulations\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e1\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e2\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e3\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e4\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e5\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e6\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. g\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e1\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e2\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.178\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e3\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.096\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.197\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e4\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.036\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.114\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e5\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.055\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.141\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.036\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e6\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.099\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.052\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.096\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.017\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.g\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.3762%;\"\u003e\n \u003cp\u003e0.174\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.225\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.128\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.221\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.208\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e0.234\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.5413%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e****\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eMutation-Drift Equilibrium\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe genetic signatures of the recent decline in the effective population size are usually characterized by reduced allelic richness and heterozygosity, with allelic richness being reduced at a faster rate, which could result in observed heterozygosity being higher than the mutation drift equilibrium (Cornuet and Luikart 1996). Bottleneck analysis was carried out using the IAM, SMM, and TPM models with Wilcoxon\u0026rsquo;s signed-rank test for heterozygosity excess and the mode-shift test to identify changes in allele frequency distribution. Among the populations surveyed, \u003cem\u003eO.e\u003c/em\u003e1 showed a change in allele frequency distribution and heterozygosity excess (p \u0026lt; 0.05) using all three models (Table 4). The bottleneck could be due to the reduced size of the effective population. Other populations showed the characteristic L-shaped curve, which is the expected curve in the absence of a bottleneck event. However, care must be taken while interpreting the results, and the presence of outbreeding depression, Hardy-Weinberg equilibrium, null alleles, and inbreeding could affect the results (Aboukhalid et al. 2017).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4\u003c/strong\u003e Summary of the Wilcoxon signed-rank test outcomes evaluating excess heterozygosity and allele frequency distribution shifts under the infinite alleles model (IAM), the stepwise mutation model (SMM), and the two-phase model (TPM). Significant deviations from mutation-drift equilibrium are indicated by \u003cem\u003ep\u003c/em\u003e-values less than 0.05.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"614\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulations\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIAM model\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTPM model\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSMM model\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMode shift\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO. e1\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.00049\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.00244\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01221\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003eShifted mode\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e2\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.16968\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.45483\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.66138\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003eNormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e3\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.58032\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.90454\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.99573\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003eNormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e4\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.47302\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.72913\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.90454\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003enormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e5\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.29395\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.55371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.79285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003enormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.e6\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.29395\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.70605\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.96594\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003enormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 21.1726%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eO.g\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.5896%;\"\u003e\n \u003cp\u003e0.84937\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0326%;\"\u003e\n \u003cp\u003e0.95386\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.7068%;\"\u003e\n \u003cp\u003e0.99329\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.4984%;\"\u003e\n \u003cp\u003enormal L- shaped\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Discussion","content":"\u003cp\u003e\u003cstrong\u003ePatterns of Genetic Diversity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSmall or narrowly distributed plant species are often susceptible to declines in genetic diversity, making their study essential for conservation planning. In this study, we assessed the genetic diversity of two Moroccan endemics, \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e, using 13 microsatellite markers. Observed heterozygosity (Ho) ranged from 0.285 in \u003cem\u003eO. grosii\u003c/em\u003e to 0.505 in \u003cem\u003eO. elongatum\u003c/em\u003e (\u003cem\u003eO.e\u003c/em\u003e5). Expected heterozygosity (He) was lower than Ho in four populations (\u003cem\u003eO.e\u003c/em\u003e1, \u003cem\u003eO.e\u003c/em\u003e2, \u003cem\u003eO.e\u003c/em\u003e4, \u003cem\u003eO.e\u003c/em\u003e5), suggesting heterozygote excess. On average, \u003cem\u003eO. elongatum\u003c/em\u003e exhibited higher genetic diversity (Ho = 0.455; He = 0.429) than \u003cem\u003eO. compactum\u003c/em\u003e from Morocco (Ho = 0.37; He = 0.35; Aboukhalid et al. 2017) and comparable values to \u003cem\u003eO. vulgare\u003c/em\u003e subsp. \u003cem\u003ehirtum\u003c/em\u003e from Bulgaria (He = 0.482; Ho = 0.448; Alekseeva et al. 2021).\u003c/p\u003e\n\u003cp\u003eIn comparison with other species of the \u003cem\u003eOriganum\u003c/em\u003e genus\u003cem\u003e, O. elongatum\u003c/em\u003e showed exceptionally high levels of genetic diversity. In \u003cem\u003eO. compactum\u003c/em\u003e, where the same SSR markers were used, the number of alleles per locus was 2.26, allelic richness was 1.72, and there were 14 private alleles (Aboukhalid et al. 2017). This study supported that low levels of genetic diversity are not universal among range-restricted species. Some range-restricted species may harbor levels of diversity comparable to or even higher than those of their common congeners (Gitzendanner and Soltis 2000). The high levels of genetic diversity found in \u003cem\u003eO. elongatum\u003c/em\u003e, despite its range-restricted nature, may indicate ecological and genetic resilience that are likely linked to broad substrate adaptability and lack of recent founder events.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eO. grosii\u003c/em\u003e showed lower levels of genetic diversity, especially regarding allelic richness compared to \u003cem\u003eO. elongatum\u003c/em\u003e. This pattern is comparable to that found for other range-restricted endemic species (Gibson et al. 2008). Confined to a single locality\u003cem\u003e, O. grosii\u003c/em\u003e illustrates this trend. Similar patterns of decreased genetic diversity have been recorded for other range-restricted species such as \u003cem\u003eSeseli farrenyi\u0026nbsp;\u003c/em\u003e(Garcia-Jacas et al. 2021). Although \u003cem\u003eO. grosii\u003c/em\u003e is a perennial species, and thus not subject to bottlenecks due to short life cycles, its small effective population size increases susceptibility to genetic drift, leading to greater homozygosity (Gustafsson and Sj\u0026ouml;gren-Gulve 2002). Surveys revealed no additional populations of \u003cem\u003eO. grosii\u003c/em\u003e, confirming its critical status and the need to prioritize its single population for conservation. Its heterozygote deficit may partly reflect null alleles, often overestimated in small or inbred populations (Van Oosterhout et al. 2005). Reduced genetic diversity in \u003cem\u003eO. grosii\u003c/em\u003e and \u003cem\u003eO. elongatum\u003c/em\u003e relative to widespread \u003cem\u003eO. vulgare\u003c/em\u003e is consistent with effects of drift, inbreeding, and allelic loss in small populations (Spielman et al. 2004; Alekseeva et al. 2021). However, genetic diversity also depends on life history, breeding system, gene flow, dispersal, and historical biogeography (Wang et al. 2004). For \u003cem\u003eO. grosii\u003c/em\u003e, low diversity is not explained by its perennial, allogamous nature; pollinator limitation or ecological specialization may contribute (Phillips et al. 2014). In \u003cem\u003eO. elongatum\u003c/em\u003e, heterozygote deficits in O.e2 and O.e6 may reflect mating among relatives.\u003c/p\u003e\n\u003cp\u003eGenetic differentiation between \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e is supported by species-specific alleles, with population assignment matching geographic origin even in sympatric areas like Chefchaouen. Admixed genotypes indicate past or ongoing gene flow, reflecting divergence with occasional interspecific exchange. The strong heterozygote deficit in O.e1 may result from null alleles (Arbel\u0026aacute;ez-Cort\u0026eacute;s et al. 2007), inbreeding or drift (Dodd et al. 2002), or a Wahlund effect from population structure (Dharmarajan et al. 2012).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePopulation Structure Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of the Bayesian clustering analysis revealed the presence of four genetic clusters, (K = 4). The results of the STRUCTURE analysis revealed the presence of three lineages of \u003cem\u003eO. elongatum\u003c/em\u003e and one lineage of \u003cem\u003eO. grosii\u003c/em\u003e, with admixed populations showing evidence of historical or recent gene flow. The results were further confirmed by the Principal Coordinate Analysis (PCoA), which aligned with the inferred genetic structure and population assignments.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn \u003cem\u003eO. elongatum\u003c/em\u003e, the populations \u003cem\u003eO.e\u003c/em\u003e2, \u003cem\u003eO.e\u003c/em\u003e4, \u003cem\u003eO.e\u003c/em\u003e5, and \u003cem\u003eO.e\u003c/em\u003e6 were grouped together, showing evidence of evolutionary history or recurrent gene flow, which might be attributed to historical or recent gene flow, founder effects, and expansion events. In accordance with our findings, a previous study carried out by SSR markers on \u003cem\u003eO. compactum\u003c/em\u003e from Morocco, revealed that founder effects and bottlenecks were responsible for the lower levels of genetic diversity (Aboukhalid et al. 2017).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBottleneck analysis detected evidence of a recent demographic decline only in population \u003cem\u003eO.e\u003c/em\u003e1, indicated by a mode-shifted allele frequency distribution and heterozygosity excess, although the small sample size (n = 8) may limit the robustness of this inference (Cornuet and Luikart 1996). The decline nevertheless appears plausible given the observed habitat degradation in this area.\u003c/p\u003e\n\u003cp\u003eGenetic structure mirrors ecological boundaries, likely driven by restricted gene flow (Epps and Keyghobadi, 2015). Admixture indicates past or ongoing gene flow, and \u003cem\u003eO. grosii\u003c/em\u003e remains sufficiently related to \u003cem\u003eO. elongatum\u003c/em\u003e for cross-species SSR amplification. Increasing habitat fragmentation from land-use change threatens gene flow and may accelerate genetic erosion in narrow endemics, highlighting the need to combine land-cover mapping and genetics to prioritize connectivity and conservation (Epps and Keyghobadi, 2015).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConservation Implications\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study provides the first comprehensive genetic baseline for \u003cem\u003eO. grosii\u003c/em\u003e and \u003cem\u003eO. elongatum\u003c/em\u003e, demonstrating that conserving genetic diversity is essential for long-term persistence and identifying priority populations beyond morphological and geographic criteria. Genetically diverse and distinct populations (notably \u003cem\u003eO.e\u003c/em\u003e4 and \u003cem\u003eO.e\u003c/em\u003e5) should be prioritized for in situ conservation and integrated into ex situ programs, while the widespread presence of private alleles (except \u003cem\u003eO.e\u003c/em\u003e1) enhances most populations\u0026rsquo; value for restoration and reintroduction (Frankham 2016).\u003c/p\u003e\n\u003cp\u003eFour genetic clusters were identified, supporting their management as separate Evolutionarily Significant Units (ESUs) and Management Units (MUs). While populations within clusters (e.g., \u003cem\u003eO.e\u003c/em\u003e2, \u003cem\u003eO.e\u003c/em\u003e4, \u003cem\u003eO.e\u003c/em\u003e5, \u003cem\u003eO.e\u003c/em\u003e6) may serve as mutual reinforcement sources, inter-cluster mixing should be avoided to reduce the risk of outbreeding depression (Waller 2015; Moritz 1994). Urgent genetic reinforcement and habitat restoration are needed for the genetically impoverished Boulemane population (O.e1). Ex situ conservation and use of local seed sources are advised to preserve local adaptation (Quilichini et al. 2004).\u003c/p\u003e\n\u003cp\u003eMajor threats including overgrazing, agriculture, urbanization, and illegal harvesting highlight the need for legal protection, inclusion in Key Biodiversity Areas, and alignment with IUCN assessments. Although fragmented, \u003cem\u003eO. elongatum\u003c/em\u003e maintains relatively high genetic diversity but limited gene flow, calling for targeted management. Integrating population genetics with ecological and climate modeling is essential to conserve adaptive capacity and ensure long-term viability under climate change (Hoffmann and Sgr\u0026ograve; 2021), providing a transferable framework for conserving other narrowly distributed endemic species.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study provides the first comprehensive genetic assessment of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e across Morocco, revealing clear inter- and intraspecific differentiation with key conservation implications. Despite habitat fragmentation, \u003cem\u003eO. elongatum\u003c/em\u003e maintains relatively high genetic diversity, though limited gene flow among populations calls for targeted actions. Populations in Taounate and Al Hoceima, with high diversity and unique genetic profiles, should be prioritized, while isolated, genetically poor populations like Boulemane need urgent reinforcement and habitat restoration. Four genetic clusters support their designation as separate management units for restoration programs. The findings underscore the value of integrating molecular data with conservation planning, considering geographic isolation, ecological gradients, and land-use pressures, to guide strategies that preserve evolutionary potential and adaptive resilience in Morocco\u0026rsquo;s endemic flora and other threatened species in fragile regions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eWe express our sincere gratitude to the \u0026ldquo;Service de Syst\u0026eacute;matique Mol\u0026eacute;culaire\u0026rdquo; (UMS 2700, Mus\u0026eacute;um National d\u0026apos;Histoire Naturelle, Paris, France) for providing access to their laboratories for the molecular genetic analyses. We also gratefully acknowledge the financial support provided by the Moroccan-French cooperation program (Toubkal).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eK.A., N.M., A.L. and C.A.F. designed and conceptualized the study. K.A. performed formal analyses and wrote the original draft. \u0026nbsp;K.A., M.B. and A.K. conducted fieldwork, sample collection and identified plant material. J.L. conceived the laboratory analyses and participated in DNA extraction, PCR amplification, and SSR genotyping. M.N., A.M. and G.K.G. performed statistical analyses, prepared some figures and contributed to manuscript review and editing. N.M., C.A.F. and A.L. supervised the project. N.M. and C.A.F. acquired funding and administered the project. All authors gave final approval for publication.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThis work was partially\u0026nbsp;supported by the \u0026ldquo;PHC\u0026nbsp;Toubkal\u0026rdquo; programme, funded by the French Ministry for Europe and Foreign Affairs, the French Ministry for Higher Education Research and Space (MESRE), and the Moroccan Ministry for Higher Education,Scientific Research and Innovation (MESRSI).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e The authors confirm that there are no financial conflicts or personal connections that might have influenced the research presented in this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAboukhalid K, Al Faiz C, Bakha M, Douaik A, Lamiri A, Gibernau M, Tomi F (2023) Agronomic evaluation and essential oil composition characterization of \u003cem\u003eOriganum compactum\u003c/em\u003e accessions cultivated in Morocco. 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J Hered 95:234\u0026ndash;243. https://doi.org/10.1093/jhered/esh043 \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"conservation-genetics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"coge","sideBox":"Learn more about [Conservation Genetics](https://www.springer.com/journal/10592)","snPcode":"10592","submissionUrl":"https://submission.nature.com/new-submission/10592/3","title":"Conservation Genetics","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Conservation Genetics, Endemic species, Origanum elongatum, Origanum grosii, Genetic Diversity, SSR Markers","lastPublishedDoi":"10.21203/rs.3.rs-8870849/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8870849/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOregano is a highly valued aromatic and medicinal herb in the Mediterranean, appreciated for its culinary, therapeutic, and ecological significance. Classification within the \u003cem\u003eOriganum\u003c/em\u003e genus is complicated by extensive morphological variation and frequent interspecies hybridization. In this study, we assessed the genetic diversity and population structure of \u003cem\u003eO. elongatum\u003c/em\u003e and \u003cem\u003eO. grosii\u003c/em\u003e across their full distribution in Morocco, sampling 264 individuals from seven populations and analyzing them with 13 microsatellite (SSR) markers. Despite its restricted native range, \u003cem\u003eO. elongatum\u003c/em\u003e displayed considerable genetic diversity (Ho\u0026thinsp;=\u0026thinsp;0.455, He\u0026thinsp;=\u0026thinsp;0.429), whereas \u003cem\u003eO. grosii\u003c/em\u003e showed lower diversity (Ho\u0026thinsp;=\u0026thinsp;0.285) but higher differentiation among populations. Bayesian clustering, Principal Coordinates Analysis, and STRUCTURE analysis revealed four main genetic groups largely corresponding to geographic origin and clearly separating the two species. These results indicate strong species divergence and the existence of distinct population structures, underscoring the need for species-specific conservation measures. We recommend integrating in-situ and ex-situ strategies, including habitat protection, genetic reinforcement, and germplasm preservation in gene banks. Overall, this study illustrates how genetic data can inform conservation planning for rare endemic plants in ecologically sensitive regions, providing a model applicable to other Mediterranean mountain ecosystems facing climate and land-use pressures.\u003c/p\u003e","manuscriptTitle":"Genetic diversity and differentiation of two narrowly endemic Origanum Species in Morocco: implications for conservation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-04 08:46:02","doi":"10.21203/rs.3.rs-8870849/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-26T03:42:07+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-19T17:06:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"219486434805773577388111891891803208205","date":"2026-04-08T15:58:44+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-20T19:26:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"332509583711621575565265602538883458275","date":"2026-02-27T07:12:08+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-27T06:59:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-16T14:41:13+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-16T14:36:45+00:00","index":"","fulltext":""},{"type":"submitted","content":"Conservation Genetics","date":"2026-02-13T10:37:26+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"conservation-genetics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"coge","sideBox":"Learn more about [Conservation Genetics](https://www.springer.com/journal/10592)","snPcode":"10592","submissionUrl":"https://submission.nature.com/new-submission/10592/3","title":"Conservation Genetics","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"1189e243-d44b-432b-b216-49a9c5caaee9","owner":[],"postedDate":"March 4th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-04-26T03:53:26+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-04 08:46:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8870849","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8870849","identity":"rs-8870849","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}