Genetic diversity and mating system of <Emphasis Type="Italic">Pinus brutia</Emphasis> var. <Emphasis Type="Italic">stankewiczii</Emphasis> Sukacz. in small localities of Sudak (Crimea)

The comparative analysis of genetic variation at 12 polymorphic isozyme loci, as well as the mating system, has been carried out in mature trees and their seed progenies in three small localities of Pinus brutia var. stankewiczii Sukacz. in the region between the town of Sudak and settlement of Novyi Svet in Crimea. We found that embryos maintain the same allelic diversity as parent trees; however, their observed heterozygosity, on average, is lower by 37.4%. The significant deviation of genotype distribution from the expected ratios caused by the deficiency of heterozygotes was observed at 8 out of 12 loci. The multilocus estimate of the out-crossing rate (t _m ) in populations varied from 68.9 to 94.9% with the average of 80.7%.     

Population genetics of <Emphasis Type="Italic">Cedrela fissilis</Emphasis> (Meliaceae) from an ecotone in central Brazil

Cedrela fissilis is an endangered timber species associated with seasonal forests throughout South America. We investigated a population of C. fissilis (PAN) located toward central Brazil to uncover insights on how an ecotone may have shaped the evolutionary history of this species at the local scale. PAN consisted of 18 mother trees and their 283 offspring (18 families), which were genotyped with ten microsatellite loci. We supplemented our dataset with equivalent microsatellite data from 175 specimens representing the east and west lineages of C. fissilis. An array of complementary methods assessed PAN for genetic diversity, population structure, and mating system. In PAN, the gene pool of the east lineage combined with a third (previously unidentified) lineage to form an admixture population. PAN is under inbreeding (Ho?=?0.80 and 0.74, uHe?=?0.85 and 0.82, Ap?=?1.1 and 7.1, F?=?0.06 and 0.10, for mother trees and offspring, respectively). Mother trees were predominantly outcrossing (t_m?=?0.95), with some selfing (1???t_m?=?0.05), and crossing between related individuals (t_m–t_s?=?0.07); they received pollen from few donors (N_ep?=?9). Restricted gene flow within PAN gave rise to a strong population structure, which split the 18 families into six groups. Some mother trees were reproductively isolated. Conservation perspectives are discussed.     

Genetic structure of populations of Norway spruce (<Emphasis Type="Italic">Picea abies</Emphasis> (L.) Karst. from Ukrainian Carpathians

The genetic diversity, subdivision, and differentiation of nine populations of Norway spruce (Picea abies (L.) Karst.) in Ukrainian Carpathians were studied using electrophoretic analysis of variability of enzyme systems in 346 trees aged from 80 to 150 years. Based on electrophoretic fractionation of enzymes extracted from seed endosperms in vertical slabs of 7.5% polyacrylamide gel, 20 loci of nine enzyme systems (ADH, ACP, DIA, GDH, GOT, MDH, LAP, FDH, SOD) were identified, and 71 allele variant were revealed. Each tree was heterozygous on average in 15.8% of its genes. The populations were characterized by low subdivision (F _ST = 0.017) and differentiation (D _N = 0.005). The main contribution to heterogeneity of population genetic structure was made by loci Dia-3, Lap-1, and Sod-3. Clustering and multivariate analysis revealed no observed trends in geographical or altitudinal position of the populations.     

Population genetics of the naturally rare tree <Emphasis Type="Italic">Dimorphandra wilsonii</Emphasis> (Caesalpinioideae) of the Brazilian Cerrado

Naturally rare species have a higher probability of stochastic extinction due to genetic, demographic, or environmental hazards; human disturbance may intensify these threats. Rare species may therefore be in need of short-term intervention to survive. The ecosystem with the second highest biodiversity in Brazil, the Cerrado, is suffering from fragmentation and threats to its flora. Dimorphandra wilsonii, a 30-m tall endemic tree of the Brazilian Cerrado, is listed as critically endangered; only 21 adult trees have been identified. We carried out mating system and pollen flow analyses to understand the current gene flow and limitations in the reproduction of D. wilsonii. With seven fluorescently labelled microsatellite primers, we genotyped 20 adult trees and 269 progeny from 13 mother trees. D. wilsonii displayed low levels of genetic diversity; bottleneck events are likely to have occurred (H _e ?=?0.60 and 0.29; H _o ?=?0.71 and 0.33, for adults and progeny, respectively). This species is predominantly outcrossing (t _m ?=?0.88), with some selfing (1-t _m ?=?0.12), as well as crossing between related individuals (t _m -t _s ?=?0.11). None of the studied trees was reproductively isolated; a high proportion of pollen (55 %) came from trees yet to be discovered. Two genetic clusters (Northern and Southern) were identified, with high values of genetic divergence among the Southern sites. Planting of seedlings and monitoring of seed dispersion in order to maintain the genetic diversity and genetic structure of D. wilsonii are strategies that may ensure the continuation of D. wilsonii, but this species does not seem to require reproductive intervention to remain viable.     

Gene flow among <Emphasis Type="Italic">Hancornia speciosa</Emphasis> (Apocynaceae) varieties and hybrid fitness

The fate of hybrids and the temporal and spatial dynamics of hybrid zones depend on hybrid fitness in comparison to non-hybrids. We studied cross-pollination among Hancornia speciosa varieties and compared progeny fitness in a nursery to address whether hybrid fitness differed from non-hybrids and whether maternal and paternal taxa contribute differentially to offspring fitness. This species has edible fruit pulp that is used as a raw material for candies, ice cream, and juice by small- and medium-sized enterprises in Central-West and Northeast Brazil. We genotyped 258 adults from a germplasm collection and 320 seeds using seven microsatellite loci to estimate genetic parameters and determine pollen donors. Fitness components, days to shoot, growth rate (mm/day), leaf width (mm), leaf length (mm), stem diameter (mm), and plant height (cm) were analyzed in 200 individuals. Genetic diversity and polymorphism did not differ neither between adults and progeny arrays nor among the four varieties. Genetic differentiation among varieties (F _CT?=?0.019, p?<?0.001) and among populations within varieties (F _SC?=?0.053, p?<?0.001) was significant but low. We detected mating among the four varieties, and no self-pollination was observed in parentage analysis, confirming that H. speciosa is self-incompatible with a high outcrossing rate (t _m?=?0.990, SE?=?0.007). No significant effect of heterosis or exogamic depression was detected for any fitness component, but maternal contribution significantly affected plant height.     

Genetic structure of populations and natural hybridization between <Emphasis Type="Italic">Dactylorhiza salina</Emphasis> and <Emphasis Type="Italic">D. incarnata</Emphasis> (Orchidaceae)

The results of studying the polymorphism and genetic structure of populations of D. salina and D. incarnata growing in Zabaykalsky krai and Buryatia are represented according to the data of allozyme analysis of eight genetic loci (PGI, NADHD, SKDH, GDH, PGM, DIA, ADH, and IDH). The specificity of the allelic structure of loci SKDH, PGM, and IDH is established, for which D. salina and D. incarnata reliably differ from each other. It is shown that interspecies introgressive hybrid complexes with different genetic structures were formed in Transbaikalia. Places of mass growth of D. incarnata were observed to have single plants of D. salina, the interspecies hybrids of the first and subsequent generations. Places of mass growth of D. salina were observed to contain only the hybrids that are not hybrids of the first generation. They were heterozygous not for three loci with differentiating alleles of both parents, SKDH, PGM, and IDH, but for only one of them. The degree of genetic differentiation among five populations of D. salina was on average 7.5% and that of D. incarnata was 7.1%, which in accordance with Wright’s estimation relates to mean values. The average value of FST for all studied populations of the two related species of the genus Dactylorhiza was 0.478, indicating a very high degree of genetic differentiation between D. salina and D. incarnata growing in Transbaikalia. The greatest differences between the species are for the allelic structure of loci SKDH, PGM, and IDH (FST was equal to 0.705, 0.976, and 0.762, respectively). Analysis of molecular variance (AMOVA) showed that populations of D. salina and D. incarnata in the zone where their ranges in Zabaykalsky krai and Buryatya overlap have essential differences both for the variation of alleles frequencies of eight loci (71%, d.f. = 9) and for the variability of genotypes (61%, d.f. = 9). Despite the fact that D. salina and D. incarnata explicitly share a gene flow as a result of interspecies hybridization, the genetic differentiation of populations of these related species remains at a high level.     

Genetic differentiation among natural populations of the lizard complex <Emphasis Type="Italic">Darevskia raddei</Emphasis> as inferred from genome microsatellite marking

The article presents the genetic parameters of the populations of lizards of the Darevskia raddei complex (D. raddei nairensis and D. raddei raddei) and the populations of D. valentini calculated on the basis of the analysis of variability of 50 allelic variants of the three nuclear genome microsatellite-containing loci of 83 individuals. It was demonstrated that the F_st genetic distances between the populations of D. raddei nairensis and D. raddei raddei were not statistically significantly different from the F_st genetic distances between the populations of different species, D. raddei and D. valentini. At the same time, these distances were statistically significantly higher than the F_st distances between the populations belonging to one species within the genus Darevskia. These data suggest deep divergence between the populations of D. raddei raddei and D. raddei nairensis of the D. raddei complex and there arises the question on considering them as separate species.     

Genetic differentiation of <Emphasis Type="Italic">Puccinia triticina</Emphasis> Erikss. in Russia

A collection of Puccinia triticina isolates was characterized for polymorphism of microsatellite loci and estimated for their differentiation by geographic origin. The collection included 20 isolates from the Ural region, 31 from West Siberia, 53 from Central Europe, 32 from the Northwest region, 32 from the Volga region, and 40 from the North Caucasus (24 from Dagestan and 16 from Krasnodar and Stavropol). The studied isolates were represented by 65 virulence phenotypes. In the polymorphism analysis of 18 microsatellite loci, 69 genotypes were determined. The index values of genetic distances (F _st, R _st, KB _c) between populations for microsatellite loci indicated differentiation of P. triticina isolates according to geographical origin, and they were clustered into three groups: (1) Asian, (2) European, and (3) North Caucasian. The North Caucasian isolates from Krasnodar and Stavropol regions were closer in similarity to European isolates than the Dagestan ones. Current analysis confirmed the assumption made earlier on the basis of the virulence analysis about the existence of several geographic fungi populations in Russia.     

Short-distance pollen dispersal in a protogynous Annonaceae tree species from the Brazilian Cerrado

Annona crassiflora (Annonaceae) is a protogynous beetle-pollinated savannah tree species, widely distributed in the savannahs of the Cerrado biome. Studies on the mating system and pollen dispersal of protogynous species are very scarce. Here, we used six microsatellite loci to assess the mating system and pollen dispersal of A. crassiflora in a savannah remnant in Central Brazil. We mapped and sampled leaves of 112 adult trees and collected 74 fruits from 20 mother trees (1–4 fruits per plant) to obtain the seeds used (460) for mating system and parentage analyses. Annona crassiflora has predominantly allogamous mating systems, with a high multilocus outcrossing rate (t_m?=?0.974, SE?=?0.011) that did not differ among mother trees (F?=?1.32, p?=?0.165). However, t_m–t_s was variable among seed trees, indicating that some seeds were produced by mating among relatives. Our results also showed multiple paternity within fruits. Multilocus correlation of outcrossed paternity was high (r_p?=?0.302, SE?=?0.045), indicating that for each mother tree, the probability that the same pollen donor sired two random sibs was 30.2%, and the mean number of pollen donors per mother tree was high (6.3). We detected a maximum pollen dispersal distance of 360.7 m and an average of 124.3 m (SD?=?80 m), but most pollination events (73%) occurred at shorter distances (<?160 m), indicating short-distance pollen dispersal, most likely due to the pollinator behaviour.     

Nucleotide polymorphisms of candidate genes of adaptive significance in the ural populations of <Emphasis Type="Italic">Larix sibirica</Emphasis> Ledeb.

The objectives of conservation and sustainable forest management require in depth study of genomes of woody plants and definition of their intraspecific genetic diversity. In recent years, an approach was developed based on the study of “candidate genes” that can potentially be involved in the formation of adaptive traits. In this study, we investigated nucleotide polymorphism of several adaptive candidate genes in the populations of Siberian larch (Larix sibirica Ledeb.) in the Urals. Representatives of this genus are among the most valuable and widely distributed forest tree species in Russia. From ten selected gene loci in the genome of L. sibirica, we isolated and investigated three loci, one of which (ABA-WDS) was sequenced in L. sibirica for the first time. The total length of the analyzed sequence in each individual amounted to 2865 bp. The length of locus alignment was from 360 bp to 1395 bp. In total, we identified 200 polymorphic positions. The most conservative is locus 4CL1-363, and the most polymorphic is locus sSPcDFD040B03103-274. The studied populations of L. sibirica are characterized by a high level of nucleotide polymorphism in comparison with other species and genuses (Picea, Pinus, Pseudotsuga, Abies) conifers plants (Hd = 0.896; π = 0.007; θ_W = 0.015). The most selectively neutral polymorphism (D _T =–0.997) was attributed to locus 4CL1-363, and polymorphism with high probability of adaptability (D _T =–1.807) was determined for the ABA-WDS locus. We identified 54 SNP markers, only five of which were nonsynonymous (9.26%) replacements. The average frequency of SNPs in the three studied loci of L. sibirica was one SNP in 53 bp. We detected unique SNP markers for eight populations, which could potentially be used to identify populations. Populations that are characterized by the highest number of unique SNP markers can be recommended for selection in order to preserve the gene pool of the species.     

Genetic analysis of forest species <Emphasis Type="Italic">Eugenia uniflora</Emphasis> L. through of newly developed SSR markers

Nine microsatellite loci for genetic analysis of three populations of the tropical tree Eugenia uniflora L. (pitanga or Brazilian cherry) from fragments of semideciduous forest were developed. We used the technique of building a (GA) _n and (CA) _n microsatellite-enriched library by capture with streptavidin-coated magnetic beads. We assessed the polymorphism of seven microsatellites in 84 mature trees found in three areas (Ribeirão Preto, Tambaú and São José do Rio Pardo), highly impacted by the agricultural practices, in a large region among Pardo river and Mogi-Guaçu river basins, in state of São Paulo, Brazil. All loci were polymorphic, and the number of alleles was high, ranging from 6 to 24, with a mean of 14.4. All stands showed the same high level of genetic diversity (mean H _E  = 0.83) and a low genetic differentiation (mean F _ST = 0.031), indicating that genetic diversity was higher within rather than among populations. Seven of the nine loci were highly variable, and sufficiently informative for E. uniflora. It was concluded that these new SSR markers can be efficiently used for gene flow studies.     

Population genetic diversity and geographical differentiation of MHC class II DAB genes in the vulnerable Chinese egret (<Emphasis Type="Italic">Egretta eulophotes</Emphasis>)

Major histocompatibility complex (MHC) genes are excellent markers for the study of adaptive genetic variation occurring over different geographical scales. The Chinese egret (Egretta eulophotes) is a vulnerable ardeid species with an estimated global population of 2600–3400 individuals. In this study, we sampled 172 individuals of this egret (approximately 6 % of the global population) from five natural populations that span the entire distribution range of this species in China. We examined their population genetic diversity and geographical differentiation at three MHC class II DAB genes by identifying eight exon 2 alleles at Egeu-DAB1, eight at Egeu-DAB2 and four at Egeu-DAB3. Allelic distributions at each of these three Egeu-DAB loci varied substantially within the five populations, while levels of genetic diversity varied slightly among the populations. Analysis of molecular variance showed low but significant genetic differentiation among five populations at all three Egeu-DAB loci (haplotype-based ?_ST: 0.029, 0.020 and 0.042; and distance-based ?_ST: 0.036, 0.027 and 0.043, respectively; all P < 0.01). The Mantel test suggested that this significant population genetic differentiation was likely due to an isolation-by-distance pattern of MHC evolution. However, the phylogenetic analyses and the Bayesian clustering analysis based on the three Egeu-DAB loci indicated that there was little geographical structuring of the genetic differentiation among five populations. These results provide fundamental population information for the conservation genetics of the vulnerable Chinese egret.     

Development and characterization of 12 polymorphic microsatellite loci in the sea sandwort, <Emphasis Type="Italic">Honckenya peploides</Emphasis>

Codominant marker systems are better suited to analyze population structure and assess the source of an individual in admixture analyses. Currently, there is no codominant marker system using microsatellites developed for the sea sandwort, Honckenya peploides (L.) Ehrh., an early colonizer in island systems. We developed and characterized novel microsatellite loci from H. peploides, using reads collected from whole genome shotgun sequencing on a 454 platform. The combined output from two shotgun runs yielded a total of 62,669 reads, from which 58 loci were screened. We identified 12 polymorphic loci that amplified reliably and exhibited disomic inheritance. Microsatellite data were collected and characterized for the 12 polymorphic loci in two Alaskan populations of H. peploides: Fossil Beach, Kodiak Island (n?=?32) and Egg Bay, Atka Island (n?=?29). The Atka population exhibited a slightly higher average number of alleles (3.9) and observed heterozygosity (0.483) than the Kodiak population (3.3 and 0.347, respectively). The overall probability of identity values for both populations was PID?=?2.892e^?6 and PID_sib?=?3.361e^?3. We also screened the 12 polymorphic loci in Wilhelmsia physodes (Fisch. ex Ser.) McNeill, the most closely related species to H. peploides, and only one locus was polymorphic. These microsatellite markers will allow future investigations into population genetic and colonization patterns of the beach dune ruderal H. peploides on new and recently disturbed islands.     

Genetic structure of the Russian populations of <Emphasis Type="Italic">Pyrenophora tritici-repentis</Emphasis>, determined by using microsatellite markers

The population genetic structure of plant pathogenic fungus Pyrenophora tritici-repentis was examined using microsatellite (SSR) markers. According to the geographical origin of the pathogen populations, they were designated as North Caucasian (S, 33 isolates), northwest (Nw, 39), and Omsk (Om, 43). The populations were analyzed at the nine most polymorphic SSR loci, at which 75 alleles were identified. To characterize the genetic variation within and between populations, the AMOVA algorithm as implemented in the Arlequin v. 3.5 software program was used. The number of alleles per locus ranged from 5 to 12 and their sizes varied within the range from 180 to 400 bp. The mean gene diversity at SSR loci was high for all populations (H = 0.58–0.75). The populations were considerably different in the frequencies of individual alleles of the SSR loci. Most isolates in the populations were represented by unique haplotypes. The within-population variation of the isolates at molecular markers was 86.4%; among the populations, 13.6%. Substantial interpopulation differences were found between the Om and S (F_st = 0.16) and between the Om and Nw (F_st = 0.20) populations, whereas between the S and Nw populations, these differences were small (F_st = 0.05). Thus, it was demonstrated that the population of P. tritici-repentis from Omsk oblast had the independent status of the geographical population; northwest and North Caucasian populations differed in the allelic diversity of SSR loci, and despite the low F_st value (0.05), they also belonged to independent geographical populations.     

Isolation of Polymorphic RAPD-SSR Markers from Xinjiang Arctic Grayling (<Emphasis Type="Italic">Thymallus arcticus grubei</Emphasis>) and a Test of Cross-Species Amplification

A total of 18 polymorphic microsatellite loci were isolated and characterized from RAPD products in the Xinjiang Arctic Grayling (Thymallus arcticus grubei). The number of alleles (N_a) per locus varied from 2 to 10. Observed (H_o) and expected (H_e) heterozygosities ranged from 0.64 to 0.92, and from 0.63 to 0.88, respectively. Considerable differences were found among HBH, FH and FY populations in the number of alleles, effective number of alleles, number of genotypes at all of these loci. These new RAPD-SSR markers have provided a helpful tool for genetic analyses and resources conservation of T. arcticus grubei. Five additional fish species, Amur grayling (Thymallus grubii), Taimen (Hucho taimen), Sea perch (Lateolabrax japonicus), Lenok (Brachymystax lenok) and Red seam bream (Pagrosomus major) were assessed for cross-species amplification. Three of the five species showed at least one polymorphic locus. In addition, seven loci were found to be polymorphic in at least one species.     

Evaluation of phylogenetic relationships in the genus <Emphasis Type="Italic">Mus</Emphasis> using <Emphasis Type="Italic">t</Emphasis>-specific microsatellite DNA markers

The t-complex includes a complex system of genes localized in the proximal region of chromosome 17 of house mouse Mus musculus. The results of microsatellite analysis of laboratory stocks of house mice carrying t ¹², t ^w5, t ^w12, and t ^w73 haplotypes and wild mice from natural populations of Russia (Volgograd, Rostov, Saratov oblasts, and Kalmykia), Armenia, Bulgaria, Iran, and Mongolia performed by the PCR method with the use of eight pairs of D17Mit primers (16, 21, 23, 28, 32, 57, 63, 78) are presented. These pairs of primers amplify microsatellite DNA sequences on mouse chromosome 17 in the region from 7.6 to 18.8 cM that correspond to inversions (In (17) 3.4). Each pair of primers recognized three to six variants of nucleotide sequences ranging in size from 90–120 bp (D17Mit 16) to 300–330 bp (D17Mit 57). In most cases, two variants of nucleotide sequences were detected in each individual, i. e., most individuals were heterozygous for the microsatellite loci under study. The highest similarity of the spectra of microsatellite DNA fragments was revealed in laboratory stocks of house mice carrying the t ^w5 and t ^w73 haplotypes. The spectra of animals from the Rostov and Volgograd oblasts appeard to be most similar to them. The microsatellite spectra of individuals from Iran closely resemble the spectrum of an individual from Armenia. It was demonstrated that amplified microsatellite fragments localized in the region of the t-complex can be used to identify representatives of the Mus genus from wild populations.     

Revealing of allelic polymorphism in the populations of parthenogenetic lizards <Emphasis Type="Italic">Darevskia dahli</Emphasis> (Lacertidae) using locus-specific PCR

Locus-specific PCR was used to study the genetic polymorphism in three populations of parthenogenetic lizard species Darevskia dahli. The analysis was carried at the two (GATA) _n -containing loci (Du215 and Du281) using the sample of 26 individuals. A total of eight Du215 and three Du281 allelic variants were detected. It was demonstrated that all the lizards examined were heterozygous at these loci. In 12 animals, unusual Du215 allelic variant was revealed, the origin of which was thought to be associated with different types of genomic rearrangements, or segmental duplication. The populations studied were substantially different relative to the levels of allelic polymorphism, which could be explained by different habitation conditions, leading to accumulation of mutations in noncoding genome regions.     

Analysis of Allelic Diversity and Genetic Relationships Among Cultivated Mangosteen (<Emphasis Type="Italic">Garcinia mangostana</Emphasis> L.) in Java,Indonesia Using Microsatellite Markers and Morphological Characters

The genetic variation and relationships of the mangosteen (Garcinia mangostana L.) were observed across mangosteen populations in Java, Indonesia using newly identified microsatellite loci and morphological characters. In this study, we developed an improved protocol to isolate microsatellite loci, named Selective Repeats from AFLP Sequence, by using a hybridized membrane. Twenty microsatellite loci were evaluated using 78 individuals from five mangosteen populations, and we successfully amplified four closely related Garcinia species, including G. malaccensis, G. hombroniana, G. celebica, and G. porrecta. Eight loci were monomorphic and the others were polymorphic. Sixty-nine alleles were found, with 3.491 per locus on an average. Genetic diversity (H?) was calculated with an average across loci within population (H?_S) as 0.39, an average loci across many populations (H?_T) as 0.444, and genetic differentiation (F?_ST) as 0.147. Furthermore, based on morphological characters, mangosteen individuals from four populations including Leuwiliang, Wanayasa, Puspahiang, and Kaligesing also had morphologically distinct fruit weight, rind weight, and rind thickness among populations. The study also elucidated the dispersal pattern of mangosteen in Java; the source of the genotype mangosteen in Java population was the Wanayasa population. In addition, we found evidence of tetraploidy in mangosteen. These results have potential applications in future breeding, conservation studies, and genetic assessment of mangosteen and their closely related species.     

Population genetic analysis of Euro-Arctic polar cod <Emphasis Type="Italic">Boreogadus saida</Emphasis> suggests fjord and oceanic structuring

Polar cod, Boreogadus saida, is a key species in Arctic marine ecosystems; however, its genetic population structure is largely undescribed. The population genetic structure of 472 B. saida specimens among nine locations in the north-east Atlantic was revealed using 12 microsatellite loci. Pairwise F _ST comparisons showed significant population differentiation between B. saida sampled inside fjords in Svalbard and north-east Greenland, as compared to B. saida from the shelf. The observed genetic variation was not a function of isolation by distance, and it is speculated that B. saida populations inhabiting fjords may have become reproductively isolated from shelf-dwelling B. saida during the last post-glacial recolonization.     

Hybrid plants preserve unique genetic variation in the St Helena endemic trees <Emphasis Type="Italic">Commidendrum rotundifolium</Emphasis> DC Roxb. and <Emphasis Type="Italic">C. spurium</Emphasis> (G.Forst.) DC

The island of St Helena in the South Atlantic Ocean has a rich endemic flora, with 10 endemic genera and 45 recognised endemic species. However, populations of most endemic species have undergone dramatic reductions or extinction due to over-exploitation, habitat destruction and competition from invasive species. Consequently, endemic species are likely to have lost genetic variation, in some cases to extreme degrees. Here, the entire extant wild populations and all planted trees in seed orchards, of two critically endangered species in the endemic genus Commidendrum (Asteraceae), C. rotundifolium and C. spurium, were sampled to assess levels of genetic variation and inbreeding. Six new microsatellite loci were developed from next-generation sequence data, and a total of 190 samples were genotyped. Some seed orchard trees contained alleles from both wild C. rotundifolium and C. spurium indicating they could be hybrids and that some backcrossing may have occurred. Some of these trees were more similar to C. rotundifolium than C. spurium both genetically and morphologically. Importantly, allelic variation was detected in the putative hybrids that was not present in wild material. C. rotundifolium is represented by just two individuals one wild and one planted and C. spurium by seven, therefore the seed orchard trees comprise an important part of the total remaining genetic diversity in the genus Commidendrum.