Mating System Parameters Research Articles

Equivalent mating system parameters in post‐mining and undisturbed native plant populations confirms restitution of bird‐pollinator function

Abstract The restoration of diverse self‐sustaining ecosystems requires re‐establishment of functional interactions among species. For plant communities, pollinators are usually essential for pollination, seed set and seed quality. A common assumption in ecological restoration for plants pollinated by animals is one of ‘build it and they will come’, which is rarely tested. Beyond seed set, there may be negative genetic consequences for seed quality if pollinators and their behaviour do not reflect those in reference populations. Here, we conduct an ecological genetic assessment of seed quality via mating system parameters in Lambertia multiflora (Proteaceae), a species dependent on nectar‐feeding birds for pollination. Four populations of L. multiflora in disturbed sites that were rehabilitated following mineral sands mining were compared with four native reference populations, near Eneabba, Western Australia. In each population, approximately 10 offspring from each of 10 maternal plants were genotyped with 11 highly polymorphic microsatellite markers. From these data, genetic diversity and mating system parameters were assessed, and found to be equivalent across all populations. Mean allelic diversity and heterozygosity across loci were very high. All populations were completely outcrossing with no bi‐parental inbreeding. Mean correlated paternity, sibship and effective population size estimates for restored and natural populations were not significantly different and reflected uniformly high paternal diversity and wide outcrossing. Equivalent genetic results for restored and natural reference populations indicate successful restitution of bird‐pollinator services for L. multiflora in these post‐mining rehabilitation sites. Synthesis and applications. Reviewing our results with other published studies to date suggests a resilience of bird‐pollinator services in restored plant communities. These findings provide some reassurance to restoration practitioners working in these global south systems where bird pollination is a feature, at least for similar landscape scenarios. Our study also highlights the global contribution of ecological genetics to the objective assessment of functional species interactions in ecological restoration, an increasingly important goal of land managers and regulators seeking to improve restoration standards.

Pollinator foraging tactics have divergent consequences for the mating system of a tropical plant.

Resolving the consequences of pollinator foraging behaviour for plant mating systems is a fundamental challenge in evolutionary ecology. Pollinators may adopt particular foraging tactics: complete trapline foraging (repeated movements along a fixed route), sample-and-shift trapline foraging (a variable route that incorporates information from previous experiences) and territorial foraging (stochastic movements within a restricted area). Studies that integrate these pollinator foraging tactics with plant mating systems are generally lacking. We investigate the consequences of particular pollinator foraging tactics for Heliconia tortuosa. We combine parentage and sibship inference analysis with simulation modelling to: estimate mating system parameters; infer the foraging tactic adopted by the pollinators; and quantify the impact of pollinator foraging tactics on mating system parameters. We found high outcrossing rates, ubiquitous multiple paternity and a pronounced departure from near-neighbour mating. We also found that plants repeatedly receive pollen from a series of particular donors. We infer that the pollinators primarily adopt complete trapline foraging and occasionally engage in sample-and-shift trapline foraging. This enhances multiple paternity without a substantial increase in near-neighbour mating. The particular pollinator foraging tactics have divergent consequences for multiple paternity and near-neighbour mating. Thus, pollinator foraging behaviour is an important driver of the ecology and evolution of plant mating systems.

Plant mating system dynamics in restoration: a comparison of restoration and remnant populations ofHakea laurina(Proteaceae)

Ecological restoration is addressing the challenge of biodiversity conservation in landscapes where native vegetation has been extensively cleared. Reestablishing ecological interactions that support self‐sustainable populations plays an essential role in restoration efforts. For animal‐pollinated plant species, comparing mating system dynamics within restoration populations with natural remnant populations can inform the progress of restoration activities. We assessed mating system parameters, seed weight, invertebrate floral visitors, and genetic diversity for two restoration populations and two native reference remnant populations of the animal‐pollinated, woody shrub/tree,Hakea laurina(Proteaceae) in the Southwest Australian Floristic Region. There was no difference in outcrossing rates between the two sites (restoration and reference populations combined) or treatments (sites combined). However, one restoration population showed significantly lower outcrossing that was correlated with reduced seed weight, greater inbreeding, and significantly lower invertebrate richness and abundance compared to its nearby remnant population. In contrast, in the other restoration population, all measures were comparable to the reference remnant population. Local genetic diversity available in remnant populations was captured in both restoration populations. Limitations to pollinator services may be affecting mating patterns and potentially population fitness, in a younger restoration population with reduced proximity to remnant vegetation, which has a tall coplanted overstorey species and spatially aggregated individuals. We highlight these aspects of restoration populations as important areas of ongoing research that will impact the reestablishment of ecological interactions, and the assessment of mating system dynamics as a valuable tool to inform the current progress of restoration activities.

The mating system affects the temperature sensitivity of male and female fertility

Abstract To mitigate the effects of climate change, it is important to understand species' responses to increasing temperatures. This has often been done by studying survival or activity at temperature extremes. Before such extremes are reached, however, effects on fertility may already be apparent. Sex differences in the thermal sensitivity of fertility (TSF) could impact species persistence under climate warming because female fertility is typically more limiting to population growth than male fertility. However, little is known about sex differences in TSF. Here we first demonstrate that the mating system can strongly influence TSF using the seed beetle Callosobruchus maculatus. We exposed populations carrying artificially induced mutations to two generations of short‐term experimental evolution under alternative mating systems, manipulating the opportunity for natural and sexual selection on the mutations. We then measured TSF in males and females subjected to juvenile or adult heat stress. Populations kept under natural and sexual selection had higher fitness, but similar TSF, compared to control populations kept under relaxed selection. However, females had higher TSF than males, and strikingly, this sex difference had increased over only two generations in populations evolving under sexual selection. We hypothesized that an increase in male‐induced harm to females during mating had played a central role in driving this evolved sex difference, and indeed, remating under conditions limiting male harassment of females reduced both male and female TSF. Moreover, we show that manipulation of mating system parameters in C. maculatus generates intraspecific variation in the sex difference in TSF equal to that found among a diverse set of studies on insects. Our study provides a causal link between the mating system and TSF. Sexual conflict, (re)mating rates and genetic responses to sexual selection differ among ecological settings, mating systems and species. Our study therefore also provides mechanistic understanding for the variability in previously reported TSFs which can inform future experimental assays and predictions of species responses to climate warming. A free Plain Language Summary can be found within the Supporting Information of this article.

Isolation and characterization of microsatellites markers in Centaurium grandiflorum ssp. boissieri.

Estimating outcrossing/selfing rates and characterizing genetic diversity with microsatellite markers are crucial to understanding the evolution of plant mating systems. We developed, optimized and characterized eight new primer pairs for Centaurium grandiflorum ssp. boissieri and transferred them to three subspecies of Centaurium quadrifolium. Two SSR loci were transferred from Sabatia campestris to the four Centaurium taxa. Polymorphisms, He, Ho and H-W deviations were estimated in two populations of C. grandiflorum ssp. boissieri and in seven individuals each of C. quadrifolium ssp. barrelieri, C. quadrifolium ssp. parviflorum and C. quadrifolium ssp. quadrifolium. A total of 80 individuals was used in these experiments. The number of polymorphic loci varied among species from one to ten. A total of 127 alleles was scored. The average number of alleles per locus was 12.7. He was higher than Ho in all sampled populations. Hardy-Weinberg equilibrium was found for some loci in different species. This is the first report of microsatellites successfully amplified in the whole Centaurium genus. They will be valuable for estimating mating system parameters and genetic diversity and exploring their relationships with the wide variation in flower morphology in the genus, especially anther-stigma separation.

Genetic and mating system assessment of translocation success of the long‐lived perennial shrub Lambertia orbifolia (Proteaceae)

There is an increasing need to establish populations of threatened plants in threat‐free habitat to prevent species extinction. The amount of genetic diversity in founding plants will influence whether the new population has the capacity to persist and evolve over time, and factors that influence the maintenance of genetic diversity, such as the mating systems, will also play a role in population persistence. We developed 13 nuclear microsatellite markers and used these to evaluate genetic diversity and mating system parameters of three translocated populations of two subspecies of Lambertia orbifolia, and compared these parameters to seven wild populations. Genetic diversity was maintained in the translocated population of L. orbifolia subsp. Scott River Plains, established using a single source population (Nar = 3.270 and He = 0.478 in translocated population; Nar = 3.280 and He = 0.534 in wild populations), and maintained or increased in the two admixed translocated populations of L. orbifolia subsp. orbifolia (Nar = 3.115, 3.830 and He = 0.511, 0.635 in translocated populations; Nar = 2.708 and He = 0.438 in wild populations) compared to wild populations of each subspecies. Mating system parameters were comparable between translocated and wild populations of L. orbifolia subsp. Scott River Plains indicating the likelihood of genetic diversity being maintained in future generations. However, there was increased selfing in translocated populations of L. orbifolia subsp. orbifolia, suggesting suboptimal pollination and high values for the inbreeding coefficient in these admixed populations (Fis = 0.474, 0.275), which may be an artifact of the Wahlund effect or from less fit (inbred) seedlings surviving ex situ propagation and translocation.

Unexpectedly low paternal diversity is associated with infrequent pollinator visitation for a bird-pollinated plant.

The behaviour of pollinators has important consequences for plant mating. Nectar-feeding birds often display behaviour that results in more pollen carryover than insect pollinators, which is predicted to result in frequent outcrossing and high paternal diversity for bird-pollinated plants. We tested this prediction by quantifying mating system parameters and bird visitation in three populations of an understory bird-pollinated herb, Anigozanthos humilis (Haemodoraceae). Microsatellite markers were used to genotype 131 adult plants, and 211 seeds from 23 maternal plants, from three populations. While outcrossing rates were high, estimates of paternal diversity were surprisingly low compared with other bird-pollinated plants. Despite nectar-feeding birds being common at the study sites, visits to A. humilis flowers were infrequent (62 visits over 21,552 recording hours from motion-triggered cameras, or equivalent to one visit per flower every 10days), and the majority (76%) were by a single species, the western spinebill Acanthorhynchus superciliosus (Meliphagidae). Pollen counts from 30 captured honeyeaters revealed that A. humilis comprised just 0.3% of the total pollen load. For 10 western spinebills, A. humilis pollen comprised only 4.1% of the pollen load, which equated to an average of 3.9 A. humilis pollen grains per bird. Taken together, our findings suggest that low visitation rates and low pollen loads of floral visitors have led to the low paternal diversity observed in this understory bird-pollinated herb. As such, we shed new light on the conditions that can lead to departures from high paternal diversity for plants competing for the pollination services of generalist nectar-feeding birds.

Evaluating restoration outcomes through assessment of pollen dispersal, mating system, and genetic diversity

Ecological genetics can provide a novel contribution to assessing the achievement of restoration objectives. We used paternity assignment to infer realized pollen dispersal within, and pollen immigration into, a restoration population of Hakea nitida, a common near‐coastal shrub or small tree in southwest Australia. We compared mating system parameters and genetic diversity with a nearby remnant reference population and assessed genetic divergence among the restoration and reference populations. We found realized pollen dispersal events closely tracked the frequency distributions of the distances between all plants within the restoration focus area. Mean realized pollen dispersal distance (359 m) approached the mean of the distances between all plants (407 m), far exceeding mean nearest neighbor distance (12 m). Maximum realized pollen dispersal distance (869 m) approached the maximum distance between all plants in the study area (1,033 m). Pollen immigration into the restoration study area was limited (4%). The mating system revealed moderate outcrossing rates (tm = 0.861 restoration and tm = 0.745 reference population), with significant and similar biparental inbreeding (tm − ts = 0.180, tm− ts = 0.186) but greater correlated paternity (rpm) in the restoration (0.519) than in the reference (0.188) population. Genetic divergence among the restoration and reference remnant population was moderate (FST = 0.094, DST = 0.239). Patterns of pollen dispersal and mating system parameters imply the attraction of pollinators within the restoration population as a key factor in progressing towards the establishment of self‐sustaining populations.

Warming and elevated CO2 induces changes in the reproductive dynamics of a tropical plant species

Tropical plant species are vulnerable to climate change and global warming. Since flowering is a critical factor for plant reproduction and seed-set, warming and elevated atmospheric carbon dioxide concentrations (eCO2) are crucial climate change factors that can affect plant reproductive dynamics and flowering related events in the tropics. Using a combined free-air CO2 enrichment and a free-air temperature-controlled enhancement system, we investigate how warming (+2 °C above ambient, eT) and elevated [CO2] (~600 ppm, eCO2) affect the phenological pattern, plant-insect interactions, and outcrossing rates in the tropical legume forage species Stylosanthes capitata Vogel (Fabaceae). In comparison to the control, a significantly greater number of flowers (NF) per plot (+62%) were observed in eT. Furthermore, in warmed plots flowers began opening approximately 1 h earlier (~09:05), with a canopy temperature of ~23 °C, than the control (~09:59) and eCO2 (~09:55) treatments. Flower closure occurred about 3 h later in eT (~11:57) and control (~13:13), with a canopy temperature of ~27 °C. These changes in flower phenology increased the availability of floral resources and attractiveness for pollinators such as Apis mellifera L. and visitors such as Paratrigona lineata L., with significant interactions between eT treatments and insect visitation per hour/day, especially between 09:00–10:40. In comparison to the control, the additive effects of combined eCO2 + eT enhanced the NF by 137%, while the number of A. mellifera floral visits per plot/week increased by 83% during the period of greatest flower production. Although we found no significant effect of treatments on mating system parameters, the overall mean multilocus outcrossing rate (tm = 0.53 ± 0.03) did confirm that S. capitata has a mixed mating system. The effects of elevated [CO2] and warming on plant-pollinator relationships observed here may have important implications for seed production of tropical forage species in future climate scenarios.

Optimising the conservation of genetic diversity of the last remaining population of a critically endangered shrub.

An understanding of genetic diversity and the population genetic processes that impact future population viability is vital for the management and recovery of declining populations of threatened species. Styphelia longissima (Ericaceae) is a critically endangered shrub, restricted to a single fragmented population near Eneabba, 250 km north of Perth, Western Australia. For this population, we sought to characterize population genetic variation and its spatial structure, and aspects of the mating portfolio, from which strategies that optimize the conservation of this diversity are identified. A comprehensive survey was carried out and 220 adults, and 106 seedlings from 14 maternal plants, were genotyped using 13 microsatellite markers. Levels of genetic variation and its spatial structure were assessed, and mating system parameters were estimated. Paternity was assigned to the offspring of a subsection of plants, which allowed for the calculation of realized pollen dispersal. Allelic richness and levels of expected heterozygosity were higher than predicted for a small isolated population. Spatial autocorrelation analysis identified fine-scale genetic structure at a scale of 20 m, but no genetic structure was found at larger scales. Mean outcrossing rate (tm = 0.66) reflects self-compatibility and a mixed-mating system. Multiple paternity was low, where 61 % of maternal siblings shared the same sire. Realized pollen dispersal was highly restricted, with 95 % of outcrossing events occurring at 7 m or less, and a mean pollen dispersal distance of 3.8 m. Nearest-neighbour matings were common (55 % of all outcross events), and 97 % of mating events were between the three nearest-neighbours. This study has provided critical baseline data on genetic diversity, mating system and pollen dispersal for future monitoring of S. longissima. Broadly applicable conservation strategies such as implementing a genetic monitoring plan, diluting spatial genetic structure in the natural population, genetically optimizing ex situ collections and incorporating genetic knowledge into translocations will help to manage the future erosion of the high genetic variation detected.

Genome-wide genotyping estimates mating system parameters and paternity in the island species Tolpis succulenta.

The mating system has profound consequences, not only for ecology and evolution, but also for the conservation of threatened or endangered species. Unfortunately, small populations are difficult to study owing to limits on sample size and genetic marker diversity. Here, we estimated mating system parameters in three small populations of an island plant using genomic genotyping. Although self-incompatible (SI) species are known to often set some self-seed, little is known about how "leaky SI" affects selfing rates in nature or the role that multiple paternity plays in small populations. We generalized the BORICE mating system program to determine the siring pattern within maternal families. We applied this algorithm to maternal families from three populations of Tolpis succulenta from Madeira Island and genotyped the progeny using RADseq. We applied BORICE to estimate each individual offspring as outcrossed or selfed, the paternity of each outcrossed offspring, and the level of inbreeding of each maternal plant. Despite a functional self-incompatibility system, these data establish T. succulenta as a pseudo-self-compatible (PSC) species. Two of 75 offspring were strongly indicated as products of self-fertilization. Despite selfing, all adult maternal plants were fully outbred. There was high differentiation among and low variation within populations, consistent with a history of genetic isolation of these small populations. There were generally multiple sires per maternal family. Twenty-two percent of sib contrasts (between outcrossed offspring within maternal families) shared the same sire. Genome-wide genotyping, combined with appropriate analytical methods, enables estimation of mating system and multiple paternity in small populations. These data address questions about the evolution of reproductive traits and the conservation of threatened populations.

Pollen dispersal, pollen immigration, mating and genetic diversity in restoration of the southern plains Banksia

AbstractEvaluation of patterns of pollen dispersal, mating systems, population fitness, genetic diversity and differentiation in restoration and remnant plant populations can be useful in determining how well restoration activities have achieved their objectives. We used molecular tools to assess how well restoration objectives have been met for populations of Banksia media in the biodiversity hotspot of south-west Western Australia. We characterized patterns of pollen dispersal within, and pollen immigration into, two restoration populations. We compared mating system parameters, population fitness via seed weight, genetic diversity and genetic differentiation for restoration and associated reference remnant populations. Different patterns of pollen dispersal were revealed for two restoration sites that differed in floral display, spatial aggregation of founders and co-planted species. Proximity to remnant native vegetation was associated with enhanced immigration and more short-range pollen dispersal when other population variables were constant. Greater seed weights at remnant compared to restoration populations were not related to outcrossing rate. Equivalent mating system and genetic diversity parameters and low to moderate levels of genetic differentiation between restoration and remnant populations suggest pollinator services have been restored in genetically diverse restoration populations of local provenance B. media as early as four years from planting.

Genetic diversity and offspring fitness in the red and white fruit color morphs of the wild strawberry Fragaria pentaphylla

Abstract Aims Fruit color polymorphisms are widespread in plants, but what maintains them is largely unclear. One hypothesis is that some morphs are preferred by dispersers while others have higher pre- or postdispersal fitness. This leads to the prediction that fruit color morphs will differ in pre- or postdispersal fitness. Methods We compared genetic and clonal diversity, mating system, morphological traits that might be associated with resistance to freezing, and germination, survival and seed production of progeny of the red and white fruit morphs in a population of a diploid, wild strawberry, Fragaria pentaphylla, from south-central China. Important Findings The red morph was much more abundant than the white but did not show higher genetic diversity as measured by observed and effective numbers of alleles, Shannon information index, or expected or observed heterozygosities. AMOVA showed that most of the genetic variation in the population was within rather than between morphs. Morphs did not differ in mating system parameters, and no significant biparental inbreeding was found in either morph. Gene flow between two morphs was high (Nm = 6.89). Seeds of the red morph germinated about 2 days earlier and had a 40% higher rate of germination than those of the white morph, but survival of seedlings and seed production by surviving offspring did not differ between morphs. The whole postdispersal fitness of the red morph was about two times higher than that of the white morph. Red morphs had hairier petioles but not more surface wax on leaves. Overall, results showed partial evidence for difference in pre- and postdispersal fitness between fruit color morphs in F. pentaphylla. Differences in fitness independent of dispersal may thus partially account for fruit color polymorphism in all cases.

Individualized mating system estimation using genomic data.

The estimation of outcrossing rates in hermaphroditic species has been a major focus in the evolutionary study of reproductive strategies, and is also essential for plant breeding and conservation. Surprisingly, genomics has thus far minimally influenced outcrossing rate studies. In this article, we generalize a Bayesian inference method (BORICE) to accommodate genomic data from multiple subpopulations of a species. As an empirical demonstration, BORICE is applied to 115 maternal families of Mimulus guttatus. The analysis shows that low-level whole genome sequencing of parents and offspring is sufficient for individualized mating system estimation: 208 offspring (88.5%) were definitively called as outcrossed, 23 (9.8%) as selfed. After mating system parameters are established (each offspring as outcrossed or selfed and the inbreeding level of maternal plants), BORICE outputs posterior genotype probabilities for each SNP genomewide. Individual SNP calls are often burdened with considerable uncertainty and distilling information from closely linked sites (within genomic windows) can be a useful strategy. For the Mimulus data, principal components based on window statistics were sufficient to diagnose inversion polymorphisms and estimate their effects on spatial structure, phenotypic and fitness measures. More generally, mating system estimation with BORICE can set the stage for population and quantitative genomic analyses, particularly researchers collect phenotypic or fitness data from maternal individuals.

Assessment of genetic diversity and mating system of Acacia cyclops restoration and remnant populations

Acacia cyclops, or Ngaamarur, is a common coastal shrub or small tree of the southwest of Western Australia and South Australia used for restoration in these landscapes and elsewhere. Knowledge of genetic diversity and mating systems of restoration populations is often lacking but can help inform likely restoration success. We compared genetic diversity and mating system parameters at three restoration populations of A. cyclops to nearby remnant reference populations. Mean levels of genetic diversity in restoration populations (Na = 1.806, Ne = 1.560, He = 0.270, Ho = 0.359) were not significantly different from those in the reference populations (Na = 1.833, Ne = 1.574, He = 0.269, Ho = 0.352) suggesting diverse seed collections were made for restoration. Allelic genetic divergence among restoration and reference populations was low (DST ≤ 0.04) suggesting that seeds were of local provenance. Mating system parameters were similar among restoration (tm = 0.989, tm–ts = 0.010, rpm = 0.392) and reference populations (tm = 0.868, tm–ts = 0.039, rpm = 0.410), suggesting functionality of the mating system and restitution of generalist insect pollinator services in restored populations at 7, 10, and 12 years after planting. Lower levels of heterozygosity in progeny compared to adults suggest post‐germination or late‐acting selection against inbred seedlings. Evidence for selection and seedling attrition further emphasizes the benefits of collections from many individuals in well‐connected outcrossed source populations. Overall, results suggest available local genetic diversity was captured and restoration populations have mating systems equivalent to those of reference populations.

Genetic variation and outcrossing rate of <i> Dipterocarpus dyeri </i> in the Tan Phu tropical rain forest (Dong Nai), Vietnam

Dipterocarpus dyeri (Dipterocarpaceae) is widely distributed in lowland rainforests in southeastern Vietnam. Due to over- exploitation and habitat destruction in the 1980s and 1990s, this species is listed as threatened. Understanding the genetic variation and mating rate among D. dyeri population that occurs in forest patches is necessary to establish effectively conservation strategies for this species. To conserve the species in tropical forests, genetic diversity and mating rate were investigated using eight microsatellites (single sequence repeat, SSR) as markers. All of the eight loci were polymorphic. A total of 36 different alleles were observed across the loci screened. The SSR data indicated high genetic diversity (NA = 4.5; HO = 0.542 and HE = 0.667) and high inbreeding value, FIS= 0,182. The mating system parameters were determined using the mixed mating model and the results indicated high outcrossing rate (tm=0.81 and ts=0.675) and lowselfing rate (0.19). Difference of tm-ts value indicated that inbreeding contributed to selfing rate for this species in the Tan Phu tropical rain forest. The results reflected that D. dyeri habitat in this area has been restored and the number of individuals was high, about 500 individuals. However, seedlings were not found so far in this study. These results indicated the importance of conserving of the genetic resources of Dipterocarpus dyeri species in the Tan Phu rain forest. The conservation strategy should include an establishment of an ex-situ conservation site with new big population for this species from all genetic groups, which might improve its fitness under different environmental stresses.

Germany: Lanxess – iron oxide pigments

Six subspecies are currently recognized in the wild olive (Olea europaea L.): cuspidata, europaea, laperrinei, maroccana, cerasiformis, and guanchica. These occur in different natural distribution ranges in Europe, Africa, and Asia. Here we present an overview and summary of the accumulated information on the molecular diversity and gene flow in the species. Past climatic oscillations and geographical barriers have been identified to govern the pattern of genetic diversity and gene movement within and between taxa. Seed dispersal by animals and cultivar transport over vast geographical distances by man are important factors which have prevented the complete differentiation of subspecies. Available evidence has shown that subsp. cuspidata is the early diverging taxon of the olive complex. However, results were not fully consistent concerning the relationships between the other subspecies. Molecular studies using nuclear and cytoplasmic markers show that the oleasters (subsp. europaea var. sylvestris) are ancestors to the cultivated forms (subsp. europaea var. europaea). However, due to hybridization in geographical proximate regions sometimes other subspecies also have influenced the cultivars. This complex process has blurred the domestication history of the species. The wild relatives of the cultivated olive are endangered in many parts of their range, albeit being important for forest restoration. Overall, molecular studies have answered a number of important questions, but most of the genetic studies so far focused on the Mediterranean oleasters, while the widespread cuspidata has not been sufficiently investigated. Using genomic approaches to understand the pattern of genetic variation will be helpful to solve open questions. Moreover, future genetic studies should focus on quantifying the local effects of disturbance factors on genetic diversity and mating system parameters of wild olive.

The critical role of honeyeaters in the pollination of the catspaw Anigozanthos humilis (Haemodoraceae)

The pollination biology of Anigozanthos humilis (Haemodoraceae) was studied within a Banksia woodland reserve using a combination of field techniques and genetic analysis. Motion-triggered cameras were deployed on 25 flowering plants to identify visitors to flowers, quantify visitation rates and assess visitor behaviour. Entire A. humilis plants were caged to exclude potential floral visitors with six treatments: (i) bird and honey possum exclusion, allowing access by insects; (ii) bird and insect exclusion, allowing access by honey possums; (iii) total animal exclusion; (iv) open, allowing access by birds, insects and honey possums; (v) hand-pollination with cross-pollen; and (vi) hand-pollination with self-pollen. Open pollinated seed were genotyped to assess mating system parameters including outcrossing rate. From 23424 h of surveillance with camera traps, 109 visits were recorded, 106 (97%) of which were western spinebills (Acanthorhynchus superciliosus), with three visits by brown honeyeaters (Lichmera indistincta). We recorded an average of 8 (±4) visits per plant, equivalent to one visit every eleven days over the 2016 flowering season (mean = 63 days). Mean seed set per fruit was negligible (0.2–0.3 seed per fruit) for all pollination treatments except open- and hand-pollination with cross pollen. Mean seed set per fruit for hand cross-pollinated flowers (37.1 seed per fruit) was 37 times that of open pollinated flowers (1.0 seed per fruit). Outcrossing rate estimates were not significantly different to one. Our results indicate that at least for the study population and season, A. humilis was largely dependent on western spinebills for delivery of outcross pollen and resulting seed set, but visitation rates were low and pollen limitation was severe. Given the known sensitivity of western spinebills to habitat fragmentation, our results suggest flow on effects that may negatively impact on reproduction, and means that A. humilis may be particularly vulnerable to environmental changes that impact on bird pollinators.

Genetic diversity and outcrossing rate of Dipterocarpus dyeri in Ma Da lowland tropical forest (Dong Nai)

Human activities often lead to the disturbed and fragmented habitat and consequently threated species. Dipterocarpus dyeri (Dipterocarpaceae) is widely distributed in lowland rainforests in southeast Vietnam. Due to over- exploitation and habitat destruction in the 1980s and 1990s, the species is listed as threatened. Understanding the genetic variation and mating rate within D. dyeri population that occurs in forest patches is necessary to establish effectively conservation strategies for this species. To conserve the species in tropical forests, genetic diversity and mating rate were investigated on the basis of eight microsatellites (single sequence repeat, SSR). All of the eight loci were polymorphic. A total of 33 different alleles were observed across the screened loci. The SSR data indicated high genetic diversities (NA = 4.1; HO = 0.461 and HE = 0.543) and the inbreeding value was high (0.185). The mating system parameters were determined using the mixed mating model and indicated high outcrossing rates (tm = 0.884 and ts = 0.645). The differences of significant tm - ts value indicated that inbreeding contributed to selfing rate for this species in lowland tropical forests of Ma Da. This study also indicated the importance of conserving the genetic resources of Dipterocarpus dyeri species in Ma Da rainforests. Genetic conservation should derive from correlated outcrossing. The conservation strategy should be established an ex-situ conservation site with new big population for this species from all genetic groups, which might improve its fitness under different environmental stresses.

Characterization of the mating system of a native perennial tetraploid herb, Silene stellata

Nursery pollination systems can range from obligate to facultative. In a system where generalists provide substantial pollination service, an important question is whether the cost of seed predation outweighs the benefit provided by the nursery pollinator to cause the plant to evolve toward more generalized pollination. Using a facultative system native to North America, we tested whether nursery pollinator vs. strictly mutualistic generalists affect mating-system parameters of the host plant and explored the implications for long-term coevolution. We used paternity analyses with 11 microsatellite markers to characterize the mating system of Silene stellata when pollination service is primarily through the nursery pollinator Hadena ectypa and generalist moths. Our experimental population of S. stellata was predominantly outcrossing (average outcrossing rate t = 0.83), and mating-system parameters were similar between pollinator groups. We detected significant correlations in both selfing and outcrossed paternity at the fruit and maternal family level, corresponding to limited pollen dispersal (mean = 3.9 m). Among individuals, variation in anther-stigma separation was positively associated with outcrossing rate, which suggests the importance of herkogamy in preventing selfing. Correlated paternity suggests that seeds from the same fruit and/or plants are sired by a limited number of pollen donors, resulting from low pollen dispersal and potential male-male competition. The similar mating-system parameters of the two pollinator groups suggest that selection for higher outcrossing in S. stellata is likely to be through floral design rather than through increased pollinator specialization with H. ectypa.