Characterizing genetic diversity and structure of populations is essential for the effective conservation of threatened species. Orchis patens sensu lato is a narrowly distributed tetraploid species with a disjunct distribution (i.e., Northern Italy, North Africa and the Canary Islands), which is facing a severe decline. In this study, we evaluated levels of genetic diversity and population structuring using 12 new nuclear microsatellite markers. Our analyses of genetic differentiation based on multiple approaches (Structure analysis, PCA analysis, and F-statistics using the ploidy-independent Rho-index) showed that gene flow is low across the range of O. patens s.l., particularly in the Canary Islands. Clear differences in allele frequencies between Italy, Algeria and the Canary Islands underlie the genetic differentiation retrieved. Our study provides support for the recognition of O. canariensis as a sister species to O. patens and the separation of the Italian populations as a new subspecies of O. patens. Despite the high heterozygosity values found in all populations (ranging from 0.4 to 0.7), compatible with the tetraploid status of the species, small population sizes and reduced gene flow will be likely detrimental for the different populations in the long term, and we recommend immediate conservation actions to counteract further fragmentation and population decline.
The genetic structure of hybrid zones provides insight into the potential for gene flow to occur between plant taxa. Four closely related European orchid species (Orchis anthropophora, O. militaris, O. purpurea, and O. simia) hybridize when they co-occur. We aimed to characterize patterns of hybridization in O. militaris-O. purpurea, O. purpurea-O. simia, and O. anthropophora-O. simia hybrid zones using molecular and morphological data. Methods: We used 11 newly isolated nuclear microsatellites to genotype 695 individuals collected from seven hybrid zones and six allopatric parental populations in France. Geometric morphometric analysis was conducted using 15 labellum landmarks to capture the main aspects of petal shape. Results: Backcrossing was asymmetric toward O. militaris in multiple O. militaris-O. purpurea hybrid zones. Hybrids in O. purpurea-O. simia and O. anthropophora- O. simia hybrid zones were largely limited to F1 and F2 generations, but further admixture had occurred. These patterns were reflected in labellum geometric morphometric data, which correlated strongly with nuclear microsatellite data in all three species combinations. Conclusions: The coexistence of parental and admixed individuals in these Orchis hybrid zones implies they are likely to be tension zones being maintained by a balance between gene flow into the hybrid zone and selection acting against admixed individuals. The pattern of admixture in the three species combinations suggests intrinsic selection acting on the hybrids is weaker in more closely related taxa.
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