Outcrossing Species Research Articles

Experimental evolution suggests rapid assembly of the 'selfing syndrome' from standing variation in Mimulus guttatus.

Ecological and evolutionary changes are likely to occur rapidly when outcrossing populations experience pollinator loss. However, the number and identify of plant traits that will respond to this form of selection, as well as the overall predictability of evolutionary responses, remain unclear. We experimentally evolved 20 large replicate populations of Mimulus guttatus for 10 generations under three treatments: pure outcrossing, mixed mating (10% outcrossing) and pure selfing. These populations were founded from the same genetically diverse and outcrossing natural population. After 10 generations, all measured traits evolved with flower size, phenology, and reproductive traits diverging consistently among mating system treatments. Autogamy increased dramatically in the selfing treatment, but the magnitude of adaptation only becomes clear once inbreeding depression is factored out. Selfing treatment plants evolved reduced stigma-anther separation, and also exhibited declines in flower size and per-flower reproductive capacity. Flower size also declined in selfing populations but this was driven mainly by inbreeding depression and cannot be attributed to adaptation towards the selfing syndrome. Generally, the mixed mating populations evolved trait values intermediate to the fully selfing and outcrossing populations. Overall, our experimental treatments reiterated differences that have been documented in interspecific comparisons between selfing and outcrossing species pairs. Given that such contrasts involve species separated by thousands or even millions of generations, it is noteworthy that large evolutionary responses were obtained from genetic variation segregating within a single natural population.

Co-flowering richness has variable effects on pollen quantity and quality limitation in four Clarkia species.

Pollination failure occurs from insufficient pollen quantity or quality. However, the relative contributions of pollen quantity vs. quality to overall pollen limitation, and how this is affected by the co-flowering context, remain unknown for most plant populations. Here, we studied patterns of pollen deposition and pollen tube formation across populations of four predominately outcrossing species in the genus Clarkia to evaluate how the richness of co-flowering congeners affects the contribution of pollen quantity and quality to pollen limitation. We partition variation in pollen deposition and pollen tube production across individuals, populations and species to identify the main sources of variation in components of reproductive success. We further quantify the relative contribution of pollen quantity and quality limitation to the reproductive success of the four Clarkia species using piecewise regression analyses. Finally, we evaluate how variation in the number of co-flowering Clarkia species in the community affects the strength of pollen quality and quality limitation. Across all contexts, pollen deposition and the proportion of pollen tubes produced varied greatly among individuals, populations and species, and these were not always correlated. For instance, C. xantiana received the smallest pollen loads yet produced the highest proportion of pollen tubes, while C. speciosa exhibited the opposite pattern. Yet, co-flowering richness had variable effects on the strength of pollen quantity and quality limitation among populations. Specifically, breakpoint values, which are an indicator of overall pollen limitation, were two-fold higher in the four-species community compared with one- and two-species communities for two Clarkia species, suggesting that pollen limitation can increase with increasing richness of co-flowering congeners. Our results reveal a complex interplay between the quantity and quality of pollen limitation and co-flowering context that may have different evolutionary outcomes across species and populations.

Low genetic differentiation despite high habitat fragmentation in an endemic and endangered species of Iridaceae from South America: implications for conservation

Abstract To conserve threatened species effectively, it is crucial to map the genetic variation of the remaining populations. Thus, using 15 microsatellites markers, from which 10 were specially developed for this study, we investigated genetic structure and gene flow patterns of Herbertia zebrina Deble, a critically endangered species endemic from grasslands of southern Brazil. We also investigated the degree of habitat fragmentation and the impacts on levels of genetic diversity, mating system and pollinators of the species. STRUCTURE and discriminant analysis of principal components identified the existence of three genetic clusters. Populations were not isolated by distance, and genetic differentiation among populations was low (7.0%). Migration rates were also low, but no evidence of genetic bottlenecks was found. However, effective population-scaled mutation rates (Θ) were < 1, suggesting that populations could be experiencing genetic drift, but the reason remains unknown. The results indicate that measurements of habitat fragmentation were not significantly correlated with genetic diversity estimates, which tend to increase with fragment size. H. zebrina was identified as an outcrossing species and specialized pollinators, such as Chalepogenus goeldianus and Lanthanomelissa betinae were rarely observed. Our findings suggest that genetic differentiation across multiple populations within the entire geographic distribution of H. zebrina is very low and populations may struggle to adapt to the current environmental and pollination changes. However, habitat fragmentation is still too recent to detect significant impacts on the levels of genetic variation. Thus, conservation plans are necessary to avoid further declines of this species.

Pervasive conservation of intron number and other genetic elements revealed by a chromosome-level genomic assembly of the hyper-polymorphic nematode Caenorhabditis brenneri.

With within-species genetic diversity estimates that span the gambit of that seen across the entirety of animals, the Caenorhabditis genus of nematodes holds unique potential to provide insights into how population size and reproductive strategies influence gene and genome organization and evolution. Our study focuses on Caenorhabditis brenneri, currently known as one of the most genetically diverse nematodes within its genus and metazoan phyla. Here, we present a high-quality gapless genome assembly and annotation for C. brenneri, revealing a common nematode chromosome arrangement characterized by gene-dense central regions and repeat rich peripheral parts. Comparison of C. brenneri with other nematodes from the 'Elegans' group revealed conserved macrosynteny but a lack of microsynteny, characterized by frequent rearrangements and low correlation iof orthogroup sizes, indicative of high rates of gene turnover. We also assessed genome organization within corresponding syntenic blocks in selfing and outcrossing species, affirming that selfing species predominantly experience loss of both genes and intergenic DNA. Comparison of gene structures revealed strikingly small number of shared introns across species, yet consistent distributions of intron number and length, regardless of population size or reproductive mode, suggesting that their evolutionary dynamics are primarily reflective of functional constraints. Our study provides valuable insights into genome evolution and expands the nematode genome resources with the highly genetically diverse C. brenneri, facilitating research into various aspects of nematode biology and evolutionary processes.

Improving precision and accuracy of genetic mapping with genotyping‐by‐sequencing data in outcrossing species

Improving precision and accuracy of genetic mapping with genotyping‐by‐sequencing data in outcrossing species

First draft reference genome and annotation of the alternative oil species Physaria fendleri.

In the wake of increasing demand for renewable energy sources, plant-based sources including alternative oilseeds have come to the forefront of interest. Hydroxy fatty acids (HFAs), produced in a few oilseed species, are important chemical feed stocks for industrial applications. An integrated approach was taken to assemble the first draft genome of the alternative HFA producer Physaria fendleri (n = 6), an outcrossing species with high heterozygosity. Both de novo transcriptome assemblies and genome assemblies were produced with public and generated sequencing reads. Resulting intermediate assemblies were then scaffolded and patched with multiple data sources, followed by super-scaffolding onto a masked genome of Camelina laxa (n = 6). Despite a current lack of available resources for the physical mapping of genomic scaffolds of P. fendleri, topography of the genome with respect to repeat and gene content was preserved at the scaffold level and not significantly lost via super-scaffolding. Read representation, gene and genome completion statistics, and annotation results illustrated the creation of a functional draft genome and a tool for future research on alternative oil species.

Assessing the Genetic Diversity of Wild and Commercial Feijoa sellowiana Accessions Using AFLPs

Feijoa sellowiana (O. Berg) is a broadly widespread fruit tree species at a very early stage of domestication. Although appreciated for its flavored berries rich in nutrients and nutraceuticals, and as an ornamental plant, feijoa is still considered an underutilized species and little information is available about its genetic background, cultivar traceability and divergence. This study aimed to investigate, for the first time, the genetic diversity of feijoa through the application of AFLPs. Specifically, twenty cultivars from different countries and six wild types (WTs) from their area of origin (Misiones, Argentina) were analyzed. The AFLPs proved to be informative, revealing the values of the percentage of polymorphic loci (PPB), Nei’s genetic diversity (h), and the Shannon index (I) at 69.36%, 0.27, and 0.43, respectively, consistent with the average of long-lived perennial and outcrossing species. However, despite the limited number of WTs examined, the genetic variability (h) was higher (approximately 37%) within the six samples compared to cultivars. The population structure analysis identified three clusters, with WTs forming a separated cluster (III) as expected. Cultivars were divided into two clusters (I and II), with cluster I exhibiting a closer genetic proximity to WTs compared to cluster II. This finding was further confirmed using the UPGMA dendrogram based on Provesti distances. This work raised awareness of the genetic variability among the feijoa’s widespread cultivars and demonstrated that the limited genetic breeding programs over the last decades resulted in low diversity among them. Moreover, these results confirm the hypothesis that all varieties are derived from a single narrow ancestral population. The potential of this species is considerable and needs to be further investigated to exploit its peculiarities.

Genetic diversity of Sinapis arvensis L. (wild mustard) in Türkiye determined by microsatellite markers

Wild mustard (Sinapis arvensis L.) is a self-fertilizing weed species that exerts negative impacts on wheat production and herbicides are intensively used to manage it. Cross-fertilization may lead to genetic differentiation in this species. Therefore, this study investigated genetic diversity among wild mustard populations collected from wheat fields across various regions in Türkiye. Genetic variation was evaluated using 5 simple sequence repeat (SSR) markers in populations collected from 30 different locations. Populations were analyzed using UPGMA (unweighted pair group method with arithmetic mean) and principal component analysis (PCA). The mean genetic diversity (GD) and polymorphism information content (PIC) values were 0.752 and 0.844, respectively. High genetic variability was recorded among populations within geographic locations. The populations were categorized into two major groups by UPGMA. There was no apparent geographic isolation among tested populations, which displayed a high degree of variability. The primary source of this variability is thought to be the adaptability of wild mustard seeds dispersed through various methods across diverse locations. Despite being a predominantly self-pollinating species, wild mustard may also employ some cross-pollination mechanisms. In conclusion, SSR markers proved useful in determining genetic diversity in outcrossing species, especially where no prior genotypic information is available. The study suggests that genetic diversity is maintained in wild mustard populations even with rotational farming practices and intensive use of herbicides.

Early-acting inbreeding depression can evolve as an inbreeding avoidance mechanism.

Despite the potential for mechanical, developmental and/or chemical mechanisms to prevent self-fertilization, incidental self-fertilization is inevitable in many predominantly outcrossing species. In such cases, inbreeding can compromise individual fitness. Unquestionably, much of this inbreeding depression is maladaptive. However, we show that when reproductive compensation allows for the replacement of inviable embryos lost early in development, selection can favour deleterious recessive variants that induce 'self-sacrificial' death of inbred embryos. Our theoretical results provide numerous testable predictions which could challenge the assumption that inbreeding depression is always maladaptive. Our work is applicable any species that cannot fully avoid inbreeding, exhibits substantial inbreeding depression, and has the potential to compensate embryos lost early in development. In addition to its general applicability, our theory suggests that self-sacrificial variants might be responsible for the remarkably low realized selfing rates of gymnosperms with high primary selfing rates, as gymnosperms exhibit strong inbreeding depression, have effective reproductive compensation mechanisms, and cannot evolve chemical self-incompatibility.

Outcrossing mating system in natural populations of the endangered aquatic fern Ceratopteris thalictroides in China as revealed by microsatellite markers: implications for conservation

In this study, the mating system of the endangered aquatic fern Ceratopteris thalictroides in China was investigated using microsatellite markers. The estimates of the multilocus outcrossing rate (tm) and single-locus outcrossing rate (ts) were high at the species level (tm = 0.701 and ts = 0.595, respectively), indicating that C. thalictroides is a predominantly outcrossing species. Negative F (inbreeding coefficient of the maternal parents) values (-0.137) at the species level indicated an excess of heterozygotes and low inbreeding in the populations analyzed. Our investigation revealed that some of the natural and ex situ conservation populations of C. thalictroides have a prolific capacity for vegetative growth. The results showed that the homosporous pteridophyte C. thalictroides in China predominantly favors gametophytic crossing with a certain level of inbreeding along with vegetative growth. Ten populations have been established for ex situ conservation in Wuhan, China, with a survival rate of more than 90%, and mature spores were collected from them. Several factors, including high dispersal of spores and gametophytes by flowing water, high gene flow, and the wide distribution of the species in China, with the occurrence of some large populations, may have resulted in a high outcrossing rate in Chinese C. thalictroides. The high outcrossing rate of C. thalictroides showed that ex situ plantings are most suitable for enhancing possible mixing of the different populations, thus minimizing inbreeding.

OcBSA: An NGS-based bulk segregant analysis tool for outcross populations

Constructing inbred lines for self-incompatible species and species with long generation times is challenging, making the use of F1 outcross/segregating populations the main strategy for genetic studies of such species. However, there is a lack of dedicated algorithms/tools for rapid quantitative trait locus (QTL) mapping using the F1 populations. To this end, we have designed and developed an algorithm/tool called OcBSA specifically for QTL mapping of F1 populations. OcBSA transforms the four-haplotype inheritance problem from the two heterozygous diploid parents of the F1 population into the two-haplotype inheritance problem common in current genetic studies by removing the two haplotypes from the heterozygous parent that do not contribute to phenotype segregation in the F1 population. Testing of OcBSA on 1800 simulated F1 populations demonstrated its advantages over other currently available tools in terms of sensitivity and accuracy. In addition, the broad applicability of OcBSA was validated by QTL mapping using seven reported F1 populations of apple, pear, peach, citrus, grape, tea, and rice. We also used OcBSA to map the QTL for flower color in a newly constructed F1 population of potato generated in this study. The OcBSA mapping result was verified by the insertion or deletion markers to be consistent with a previously reported locus harboring the ANTHOCYANIN 2 gene, which regulates potato flower color. Taken together, these results highlight the power and broad utility of OcBSA for QTL mapping using F1 populations and thus a great potential for functional gene mining in outcrossing species. For ease of use, we have developed both Windows and Linux versions of OcBSA, which are freely available at: https://gitee.com/Bioinformaticslab/OcBSA.

Influence of the mutation load on the genomic composition of hybrids between outcrossing and self-fertilizing species.

Hybridization is a natural process whereby two diverging evolutionary lineages reproduce and create offspring of mixed ancestry. Differences in mating systems (e.g., self-fertilization and outcrossing) are expected to affect the direction and extent of hybridization and introgression in hybrid zones. Among other factors, selfers and outcrossers are expected to differ in their mutation loads. This has been studied both theoretically and empirically; however, conflicting predictions have been made on the effects mutation loads of parental species with different mating systems can have on the genomic composition of hybrids. Here, we develop a multi-locus, selective model to study how the different mutation load built up in selfers and outcrossers as a result of selective interference and homozygosity impact the long-term genetic composition of hybrid populations. Notably, our results emphasize that genes from the parental population with lesser mutation load get rapidly overrepresented in hybrid genomes, regardless of the hybrids own mating system. When recombination tends to be more important than mutation, outcrossers' genomes tend to be of higher quality and prevail. When recombination rates are low, however, selfers' genomes may reach higher quality than outcrossers' genomes and prevail in the hybrids. Taken together, these results provide concrete insights into one of the multiple factors influencing hybrid genome ancestry and introgression patterns in hybrid zones containing species with different mating systems.

Frequency of Outcrossing and Isolation Distance in Faba Beans (Vicia faba L.)

Faba beans (Vicia faba L.) constitute a partially outcrossing species requiring an isolation distance to maintain genetic purity when more than one variety is grown in field conditions. This information is crucial for seed growers and faba bean breeders. A study was conducted at the University of Sydney’s Plant Breeding Institute, Narrabri, over two years to examine the extent of natural outcrossing using a creamy white flower characteristic as a morphological marker, which is controlled by a single recessive gene. The white-flowered genotype (IX225c) was grown in paired rows of 150 m length in four directions from a central 480 m2 plot of the normal flowered genotype PBA Warda. A beehive was placed in the central plot at the flowering time and natural pollination was allowed. At maturity, seed samples were taken from the white-flowered genotype at designated intervals along each axis and 100 seeds from each sample were grown in the glasshouse/birdcage to the 4–5 leaf stage and the proportion of plants displaying a stipule spot pigmentation (normal flower color and spotted stipule are linked) was used to determine the percentage of outcrossing. Maximum outcrossing of 2.28% occurred where both genotypes were grown side by side (0 m) and the degree of outcrossing decreased as the distance along each axis from the central plot increased. At a 6 m distance, the outcrossing was less than 1%; however, on occasion, it increased to 1% beyond a distance of 100 m, indicating the volatile and unpredictable nature of bee flights. Distance had a major effect on outcrossing but the direction and its interaction had no effect. The results suggest that to limit outcrossing to below 0.5%, a distance of more than 150 m between plots of different faba beans cultivars would be required. It also indicated that Australian faba bean genotypes are mostly self-fertile and a relatively narrow isolation distance will ensure self-fertilization in seed production and breeding programs.

Complex cross-incompatibility in morning glories is consistent with a role for mating system in plant speciation.

Reproductive isolation between selfing and outcrossing species can arise through diverse mechanisms, some of which are directly associated with differences in mating system. We dissected cross-incompatibility between the highly selfing morning glory Ipomoea lacunosa and its mixed-mating sister species Ipomoea cordatotriloba. We found that cross-incompatibility is complex, with contributions acting both before and after fertilization. We then investigated whether the transition in mating system may have facilitated the evolution of these reproductive barrier components through mismatched floral morphology, differences in reproductive context, or both. We found evidence that morphological mismatch likely contributes to reproductive isolation in at least one cross-direction and that other pollen-pistil interactions are present. We also identified hybrid seed inviability consistent with the predictions of the weak-inbreeder, strong-outbreeder hypotheses, suggesting endosperm misregulation plays an important role in cross-incompatibility. In contrast, we did not find evidence consistent with the prezygotic weak-inbreeder, strong-outbreeder hypothesis. Our study highlights the complexity of reproductive isolation between outcrossing and selfing species and the extent to which evolutionary consequences of mating system transitions can facilitate speciation.

Quantifying the fraction of new mutations that are recessive lethal.

The presence and impact of recessive lethal mutations have been widely documented in diploid outcrossing species. However, precise estimates of the proportion of new mutations that are recessive lethal remain limited. Here, we evaluate the performance of Fit∂a∂i, a commonly used method for inferring the distribution of fitness effects (DFE), in the presence of lethal mutations. Using simulations, we demonstrate that in both additive and recessive cases, inference of the deleterious nonlethal portion of the DFE is minimally affected by a small proportion (<10%) of lethal mutations. Additionally, we demonstrate that while Fit∂a∂i cannot estimate the fraction of recessive lethal mutations, Fit∂a∂i can accurately infer the fraction of additive lethal mutations. Finally, as an alternative approach to estimate the proportion of mutations that are recessive lethal, we employ models of mutation-selection-drift balance using existing genomic parameters and estimates of segregating recessive lethals for humans and Drosophila melanogaster. In both species, the segregating recessive lethal load can be explained by a very small fraction (<1%) of new nonsynonymous mutations being recessive lethal. Our results refute recent assertions of a much higher proportion of mutations being recessive lethal (4%-5%), while highlighting the need for additional information on the joint distribution of selection and dominance coefficients.

Effects of Wild Forest Fires on Genetic Diversity and Population Structure of a Boreal Conifer, White Spruce (Picea glauca (Moench) Voss): Implications for Genetic Resource Management and Adaptive Potential under Climate Change

Climate change is predicted to increase forest fires in boreal forests, which can threaten the sustainability of forest genetic resources. Wildfires can potentially impact genetic diversity and population structure in forest trees by creating population bottlenecks, and influencing demography, effective population size (Ne) and various evolutionary processes. We have investigated this critical issue in a widely-distributed, transcontinental, ecologically and economically important and fire-intolerant boreal conifer, white spruce (Picea glauca (Moench) Voss). We tested the hypothesis that in a predominantly outcrossing species with long distance gene flow, such as white spruce, located in primary undisturbed forests, normal forest fires do not adversely affect genetic diversity and population structure. We used 10 nuclear genic and genomic microsatellite loci to examine genetic diversity and population structure of post-fire pristine old-growth (PF-OG) and adjacent post-fire naturally regenerated young (PF-YR) stands. The genetic diversity, inbreeding and genetic differentiation levels, Bayesian population structure, Ne and latent genetic potential were statistically similar between the PF-OG and PF-YR populations. None of the microsatellites showed any signature of selection. Our study demonstrates that normal wild forest fires do not adversely affect genetic diversity, differentiation, and population genetic structure in white spruce. The results should have wide significance for sustainable forest management.

No detectable changes in reproductive behaviour of Caenorhabditis elegans males after 97 generations under obligatory outcrossing.

In Caenorhabditis elegans, a species reproducing mostly via self-fertilization, numerous signatures of selfing syndrome are observed, including differences in reproductive behaviour compared to related obligatory outcrossing species. In this study we investigated the effect of nearly 100 generations of obligatory outcrossing on several characteristics of male reproductive behaviour. A genetically uniform ancestral population carrying a mutation changing the reproductive system to obligatory outcrossing was split into four independent populations. We predicted that the transition from the natural reproductive system, where males were extremely rare, to obligatory outcrossing, where males comprise 50% of the population and are necessary for reproduction, will increase the selection pressure on higher effectiveness of mating behaviour. Several characteristics of male mating behaviour during a 15 min interaction as well as copulation success were compared between the ancestral and evolved populations. No significant differences in male mating behaviour or fertilization success were detected between generations 1 and 97 of obligatory outcrossing populations. We found, however, that longer contact with females increased chances of successful copulation, although this effect did not differ between populations. We conclude that either selection acting on male mating behaviour has not been strong enough, or mutational input of new adaptive variants has not been sufficient to cause noticeable behavioural differences after 97 generations of evolution starting from genetically uniform population.

Floral biology, pollination vectors and breeding system of Zieria granulata (Rutaceae), an endangered shrub endemic to eastern Australia

Context Understanding the factors that influence viable-seed production is crucial in the conservation of threatened plant species, yet these factors are often poorly understood. Aims We investigated the reproductive biology of Zieria granulata C.Moore ex Benth., an endangered Australian endemic with a limited distribution, with the intent of improving conservation and restoration outcomes. Methods Components of floral biology, including floral ontogeny and nectar production, were quantified to determine the pollination syndrome and the likely breeding system. Flower-visitor surveys (using both digital video recordings and human observations), a manipulative wind-pollination experiment and hand-pollination experiments were conducted to investigate pollination vectors and confirm the breeding system. Key results Z. granulata flowers were small, white, protandrous and produced highly ornamented pollen grains and small quantities of nectar; these characteristics suggest that the species fits the general entomophily syndrome. All floral visitors were arthropod species and of the 72 visitors observed, predominantly from the Dipteran and Hymenopteran families, 18 could be regarded as potential pollinators. Failure of simulated wind gusts (40 km h−1) to transport pollen ≥5 cm indicated that anemophily is unlikely for this species. Autonomous and manipulative selfing did not result in viable seed set, indicating that this is an obligate outcrossing species. However, fruit and viable-seed production was highly variable within and among some other treatments. Pre-dispersal seed predation was recorded at all study sites. Conclusions Pre-dispersal seed predation was recorded at all study sites and is a likely factor inhibiting viable-seed production. Implications This knowledge will be used to improve seed yield for collections used for ex situ conservation and restoration programs for the endangered Z. granulata.

Evolvability and trait function predict phenotypic divergence of plant populations

Understanding the causes and limits of population divergence in phenotypic traits is a fundamental aim of evolutionary biology, with the potential to yield predictions of adaptation to environmental change. Reciprocal transplant experiments and the evaluation of optimality models suggest that local adaptation is common but not universal, and some studies suggest that trait divergence is highly constrained by genetic variances and covariances of complex phenotypes. We analyze a large database of population divergence in plants and evaluate whether evolutionary divergence scales positively with standing genetic variation within populations (evolvability), as expected if genetic constraints are evolutionarily important. We further evaluate differences in divergence and evolvability-divergence relationships between reproductive and vegetative traits and between selfing, mixed-mating, and outcrossing species, as these factors are expected to influence both patterns of selection and evolutionary potentials. Evolutionary divergence scaled positively with evolvability. Furthermore, trait divergence was greater for vegetative traits than for floral (reproductive) traits, but largely independent of the mating system. Jointly, these factors explained ~40% of the variance in evolutionary divergence. The consistency of the evolvability-divergence relationships across diverse species suggests substantial predictability of trait divergence. The results are also consistent with genetic constraints playing a role in evolutionary divergence.

Genetic diversity in gooseberry (Ribes uva-crispa), as estimated with SSR markers

• Two hundred forty two gooseberry accessions were analysed by 15 SSR markers enabling the identification of 153 unique genotypes. • The genetic diversity of the analysed accessions indicated a slightly negative assortative mating as expected for an outcrossing species. • Cultivars that have been wide spread in cultivation in sweden had a large number of synonymous samples. • The studied genotypes clustered into four posterior groups corresponding to the three a priori anticipated groups: old cultivars, modern cultivars, eastern cultivars, and an additional northern swedish cluster. • Pedigrees for a number of cultivars were confirmed and supported the true-to-typeness of the involved accessions. Additionally three likely parent-parent-offspring relationships were identified. European gooseberry ( Ribes uva-crispa L. ) is a popular berry crop in many European countries, including Sweden, Denmark and Ukraine. There is no active gooseberry breeding programme in either Sweden or Denmark, but a successful programme is operating in Ukraine. In Sweden and Denmark, research on gooseberries is primarily focused on collection and phenotypic evaluation of genetic resources. As part of these activities, a large number of inventory finds have been collected but have not yet been characterised morphologically or molecularly. The goal of this study was thus to characterise gooseberry germplasm with 15 simple sequence repeat (SSR) markers. From 242 accessions analysed, 153 unique genotypes were identified. Cultivars that have been in widespread cultivation in Sweden, such as the Finnish cultivars ‘Hinnonmäen Keltainen’ and ‘Hinnonmäen Punainen’, had relatively large numbers of synonymous samples. While many inventory finds were identifiable as synonyms of known cultivars, several were found to constitute unique genotypes within the germplasm studied. The studied genotypes clustered relatively well in three posterior groups, consisting of cultivars originating before and after the American gooseberry mildew ( Sphaerotheca mors-uvae ) outbreak around 1900 and cultivars originating from the territory of the former Soviet Union. A fourth genetic cluster consisting mainly of inventory finds from central and northern Sweden was also identified. In addition, it was possible to verify recorded and stipulated parentages for some of the cultivars studied and to identify three likely parent-parent-child trios. Thus, inventories of local gooseberry germplasm and a subsequent genotyping proved successful in finding unique local genotypes, with potential local adaptation. The data obtained provide a foundation for future studies of gooseberry genetic resources, while also illustrating the importance of a well-curated and phenotypically characterised set of reference cultivars for future studies.