Contrasting genetic structure, mode of reproduction and clonal characteristics in established and newly colonised populations of Carex salina Wahlenb. (Cyperaceae)

BackgroundThe reasons that clonal plants shift between sexual and clonal reproduction have persisted as a knowledge gap in ecological literature. We hypothesized that clonal plants’ shifts between sexual and clonal reproduction in different environments are driven by the relative costs of sexual and clonal reproduction. Moreover, we hypothesized plants prioritize sexual reproduction over clonal reproduction. To test these hypotheses, we determined the costs of sexual and clonal reproduction, and proportions of sexual and clonal reproduction of Caragana stenophylla along a climatic aridity gradient (semi-arid, arid, very arid and intensively arid zones) in the Inner Mongolia Steppe using several complementary field experiments.ResultsThe cost of sexual reproduction increased while the cost of clonal reproduction decreased as climatic drought stress increased from the semi-arid to the intensively arid zones. The changes in the costs of these reproductive modes drove a shift in the reproductive mode of C. stenophylla from more sexual reproduction in the semi-arid zone to more clonal propagation in the intensively arid zone. However, because of the evolutionary advantages of sexual reproduction, sexual reproduction still held priority over clonal production in C. stenophylla, with the priority of sexual reproduction gradually increasing from the semi-arid to the intensively arid zones.ConclusionsOur study suggested that sexual reproduction has relatively high priority in propagation of C. stenophylla. However, if the costs of sexual reproduction are too high, C. stenophylla likely chooses clonal reproduction, and the ratio between sexual and clonal reproduction could be mediated by reproductive cost. These reproductive strategies reflect optimal resource utilization, and allow the persistence of both reproductive modes across stressful conditions depending on their evolutionary advantages.

Over the last 10 years plant pathologists have begun to realize that more knowledge about the genetic structure of populations of plant pathogens is needed to implement effective control strategies (48). Research on the genetic structure of fungal populations has mushroomed, and review papers that summarize these studies are numerous (7,27,33,34,38). Although the number of fungal studies has increased greatly, the most comprehensive work has focused on a small number of plant-pathogenic fungi. The majority of these fungi can be recognized easily by their fruiting bodies or disease symptoms on aboveground plant parts. It has proven more difficult to assess the genetic structure of fungal populations that exist mainly belowground, because the distribution of individuals cannot be visualized directly and appropriate sampling procedures are less obvious and more cumbersome. Nevertheless, substantial progress has been made in interpreting the population genetic structure of some soilborne fungi (1,17). The purpose of this paper is to provide an overview of the tools and techniques of fungal population genetics. I will try to emphasize approaches that may be applied to studies of soilborne fungi. Instead of providing detailed methods, I will cite recent references where appropriate. There are many opinions regarding which techniques and tools are best suited to studies of fungal populations. I will give a personal and biased viewpoint, which I believe will be most useful to those who are just entering the field.

Random genetic drift, natural selection and restricted gene dispersal are basic factors of the spatial genetic structure (SGS) in plant populations. Clonal reproduction has a profound effect on population dynamics and genetic structure and thus emerges as a potential factor in contributing to and modelling SGS. In order to assess the impact of clonality on SGS we studied clonal structure and SGS in the population of Populus alba. Six hundred and seventy-two individuals were mapped and genotyped with 16 nuclear microsatellite markers. To answer the more general question regarding the relationship between SGS and clonality we used Sp statistics, which allows for comparisons of the extent of SGS among different studies, and the comparison of published data on SGS in clonal and non-clonal tree species. Sp statistic was extracted for 14 clonal and 27 non-clonal species belonging to 7 and 18 botanical families, respectively. Results of genetic investigations conducted in the population of P. alba showed over-domination of clonal reproduction, which resulted in very low clonal diversity (R = 0.12). Significant SGS was found at both ramet (Sp = 0.095) and genet level (Sp = 0.05) and clonal reproduction was indicated as an important but not sole driving factor of SGS. Within-population structure, probably due to family structure also contributed to high SGS. High mean dominance index (D = 0.82) indicated low intermingling among genets. Literature survey revealed that clonal tree species significantly differ from non-clonal species with respect to SGS, having 2.8-fold higher SGS. This led us to conclude that clonality is a life-history trait that can have deep impact on processes acting in populations of clonal tree species leading to significant SGS.

Social Hybridogenesis in the Clonal Ant Cataglyphis hispanica

The relative importance of sexual and clonal reproduction for population growth in clonal plants is highly variable. Clonal reproduction is often more important than sexual reproduction but there is considerable interspecific variation and the importance of the two reproductive modes can change with environmental conditions. We carried out a demographic study on the woodland strawberry ( Fragaria vesca ), a widespread clonal herb, at 12 sites in Switzerland during 2 years. Study sites were selected in two different habitats, i.e., forest and forest edge. We used periodic matrix models to estimate annual population growth rates and carried out prospective analyses to identify life cycle components that influence population growth rates most. Retrospective analyses were applied to study how the two different habitats affected population dynamics. Furthermore, we tested whether trade‐offs between sexual and clonal reproduction occurred. There were large differences in annual population growth rates between sites and large within‐site differences between years. Results of the prospective analyses clearly indicate that clonal reproduction is the dominant reproductive pathway whereas sexual reproduction is rather insignificant for population growth. Compared to forest habitats, forest edge habitats had higher population growth rates in the first year but smaller growth rates in the second year. We attribute these differing habitat effects to different water availabilities during consecutive years. No trade‐offs between sexual and clonal reproduction were found. In conclusion, population growth of F. vesca relies heavily on clonal reproduction. Furthermore, reproduction and survival rates of F. vesca depend highly on spatio‐temporal variation of environmental conditions.

Aquatic plants share a range of convergent reproductive strategies, such as the ability to reproduce both sexually and asexually through vegetative growth. In dryland river systems, floodplain inundation is infrequent and irregular, and wetlands consist of discrete and unstable habitat patches. In these systems, life history strategies such as long-distance dispersal, seed longevity, self-fertilisation, and reproduction from vegetative propagules are important strategies that allow plants to persist. Using two aquatic plants, Marsilea drummondii and Eleocharis acuta, we investigated the proportions of sexual and asexual reproduction and self-fertilisation by employing next-generation sequencing approaches, and we used this information to understand the population genetic structure of a large inland floodplain in western New South Wales (NSW), Australia. Asexual vegetative reproduction and self-fertilisation were more common in M. drummondii, but both species used sexual reproduction as the main mode of reproduction. This resulted in a highly differentiated genetic structure between wetlands and a similar genetic structure within wetlands. The similarity in genetic structure was influenced by the wetland in the two species, highlighting the influence of the floodplain landscape and hydrology on structuring population genetic structure. The high levels of genetic variation among wetlands and the low variation within wetlands suggests that dispersal and pollination occur within close proximity and that gene flow is restricted. This suggests a reliance on locally sourced (persistent) seed, rather than asexual (clonal) reproduction or recolonisation via dispersal, for the population maintenance of plants in dryland rivers. This highlights the importance of floodplain inundation to promote seed germination, establishment, and reproduction in dryland regions.

Summary 1. The spatial distribution of individual plants within a population and the population’s genetic structure are determined by several factors, like dispersal, reproduction mode or biotic interactions. The role of interspecific interactions in shaping the spatial genetic structure of plant populations remains largely unknown. 2. Species with a common evolutionary history are known to interact more closely with each other than unrelated species due to the greater number of traits they share. We hypothesize that plant interactions may shape the fine genetic structure of closely related congeners. 3. We used spatial statistics (georeferenced design) and molecular techniques (ISSR markers) to understand how two closely related congeners, Thymus vulgaris (widespread species) and T. loscosii (narrow endemic) interact at the local scale. Specific cover, number of individuals of both study species and several community attributes were measured in a 10 × 10 m plot. 4. Both species showed similar levels of genetic variation, but differed in their spatial genetic structure. Thymus vulgaris showed spatial aggregation but no spatial genetic structure, while T. loscosii showed spatial genetic structure (positive genetic autocorrelation) at short distances. The spatial pattern of T. vulgaris’ cover showed significant dissociation with that of T. loscosii. The same was true between the spatial patterns of the cover of T. vulgaris and the abundance of T. loscosii and between the abundance of each species. Most importantly, we found a correlation between the genetic structure of T. loscosii and the abundance of T. vulgaris: T. loscosii plants were genetically more similar when they were surrounded by a similar number of T. vulgaris plants. 5. Synthesis. Our results reveal spatially complex genetic structures of both congeners at small spatial scales. The negative association among the spatial patterns of the two species and the genetic structure found for T. loscosii in relation to the abundance of T. vulgaris indicate that competition between the two species may account for the presence of adapted ecotypes of T. loscosii to the abundance of a competing congeneric species. This suggests that the presence and abundance of close congeners can influence the genetic spatial structure of plant species at fine scales.

Reproductive Ecology of Tetraphis pellucida. I. Population Density and Reproductive Mode

Clonality is a widespread life history trait in flowering plants that may be essential for population persistence, especially in environments where sexual reproduction is unpredictable. Frequent clonal reproduction, however, could hinder sexual reproduction by spatially aggregating ramets that compete with seedlings and reduce inter‐genet pollination. Nevertheless, the role of clonality in relation to variable sexual reproduction in population dynamics is often overlooked. We combined population matrix models and pollination experiments to compare the demographic contributions of clonal and sexual reproduction in three Dicentra canadensis populations, one in a well‐forested landscape and two in isolated forest remnants. We constructed stage‐based transition matrices from 3 years of census data to evaluate annual population growth rates, λ. We used loop analysis to evaluate the relative contribution of different reproductive pathways to λ. Despite strong temporal and spatial variation in seed set, populations generally showed stable growth rates. Although we detected some pollen limitation of seed set, manipulative pollination treatments did not affect population growth rates. Clonal reproduction contributed significantly more than sexual reproduction to population growth in the forest remnants. Only at the well‐forested site did sexual reproduction contribute as much as clonal reproduction to population growth. Flowering plants were more likely to transition to a smaller size class with reduced reproductive potential in the following year than similarly sized nonflowering plants, suggesting energy trade‐offs between sexual and clonal reproduction at the individual level. Seed production had negligible effects on growth and tuber production of individual plants. Our results demonstrate that clonal reproduction is vital for population persistence in a system where sexual reproduction is unpredictable. The bias toward clonality may be driven by low fitness returns for resource investment in sexual reproduction at the individual level. However, chronic failure in sexual reproduction may exacerbate the imbalance between sexual and clonal reproduction and eventually lead to irreversible loss of sex in the population.

Adjusting relative allocation between sexual and clonal reproduction is part of how plants respond to a variable environment, but we know little about how the allocation strategies of plant populations vary over time as abiotic conditions change. We studied correlations between sexual and clonal reproduction using 23 years of data on the clonal salt marsh plant Spartina alterniflora at eight sites in coastal Georgia, USA. The relationship between sexual reproduction and clonal reproduction varied over time. Within years, sexual reproduction was negatively related to clonal reproduction. These relationships were stronger in cooler years and weaker in warmer years, with slopes ranging from -0.202 in cool years to -0.013 in warm years. The trade-offs were also affected by river discharge, with stronger (more negative) slopes as river discharge increased. In a random forest model, temperature had the greatest influence (58%) on the relationships compared to other global change variables (precipitation, river discharge, sea level, and tide range). Overall, our study demonstrated that climate warming gradually disrupts the negative correlation in allocation between reproductive modes in a common salt marsh plant, shifting the affected populations toward a near-total reliance on clonal reproduction, potentially limiting their spread and the generation of new genotypes.

<p>Understanding the evolutionary forces that shape populations in the marine environment is critical for predicting population dynamics and dispersal patterns for marine organisms. For organisms with complex reproductive strategies, this remains a challenge. Sponges fulfil many functional roles and are important components of benthic environments in tropical, temperate and polar oceans. They have evolved diverse reproductive strategies, reproducing both sexually and asexually, and thus provide an opportunity to investigate complicated evolutionary questions. This PhD thesis examines sexual and asexual reproduction in two common golf-ball sponges in central New Zealand (Tethya bergquistae and T. burtoni), with particular focus on how the environment influences these modes of reproduction, and further, how they shape species delineations and connectivity patterns. New Zealand waters are projected to experience increases in temperature and decreases in nutrients over the next century, and therefore these species may be experience changes in basic organismal processes like reproduction due to climate change, requiring adaptation to local environments. Therefore, this work has important implications when considering how reproductive phenology, genetic diversity and population structure of marine populations may change with shifts in climate. In my first data chapter, I highlight the difficulty in delineating sponge species by investigating the evolutionary relationship of Tethya spp. in central New Zealand using both morphological and molecular methods. Phylogenetic reconstructions based on two mitochondrial markers (rnl, COI-ext) and one nuclear marker (18S) revealed three genetic clades, with one clade representing T. bergquistae and two clades belonging to what was a priori thought to be a single species, T. burtoni. Morphological analysis based on spicule characteristics allowed T. bergquistae to be distinguished from T. burtoni, but revealed no apparent differences between the T. burtoni clades. These results indicate hidden genetic diversity within T. burtoni, which likely represents a group consisting of incipient species that have undergone speciation but have yet to express clear morphological differences. This chapter supports the notion that cryptic speciation in sponges may go undetected and diversity underestimated when using only morphology-based taxonomy, a result which has implications for conservation and management of marine systems. In my second data chapter, I characterize the reproductive biology for both species of Tethya in relation to potential environmental drivers, including sea surface temperature, chlorophyll-a concentration and rainfall. Using histological methods for sponges collected monthly over two years, Tethya spp. were found to be gonochoristic and oviparous sexual reproducers, with one annual reproductive event occurring in the austral summer from January to March. Differences in oocyte density and reproductive output between both species and sites highlighted both species-specific adaptive responses and environmental influences on reproduction. Temperature and rainfall were found to be correlated with instances of sexual reproduction, and the summer reproductive event occurred each year following the spring bloom of chlorophyll-a. These findings indicate that seasonal fluctuations in the environment may be important for triggering gametogenesis for these species. With shifts in temperature, productivity, and timing of seasons projected for New Zealand, there is a potential for reproductive phenology to become mismatched with the surrounding environment under future climate change scenarios, which has consequences for the frequency, duration and overall output of sexual reproduction for these sponges. My third data chapter characterizes asexual reproduction in both species of Tethya, exploring relationships between reproductive traits and potential environmental drivers that may influence asexual budding events. Two sponge populations, one for each species of Tethya, were monitored over two years by both monthly sampling and periodic in situ observations. Data revealed that budding occurred continuously throughout the year, but had a cyclic pattern where instances of budding and densities of buds were higher during the austral spring and summer. Asexual reproduction coincided with sexual reproduction, and some individuals were found to simultaneously reproduce using both modes. Instances of asexual reproduction were positively associated with temperature and rainfall, but distinct differences between species were difficult to identify. As temperature proved important, an experiment looking at bud production in relation to thermal stress was conducted, where sponges were subjected to stable temperatures treatments of 17°C (control), 19°C and 21°C. No instances of budding were observed under any temperature treatment, and high mortality occurred in the 21°C treatment. These results suggest that temperature changes (i.e., heterogeneous environments) may be more important than temperature alone in driving asexual reproduction, and further, indicate thermal stress will result in increased sponge mortality. Correlations to potential environmental drivers indicate that future shifts in climate may affect instances of asexual reproduction and thus sponge abundance, which has the potential to alter the genetic structure and overall diversity of these populations. In the final data chapter, I developed novel microsatellite markers for Tethya burtoni to characterize the genetic connectivity patterns among four populations in central New Zealand, with particular interest in the roles that sexual and asexual reproduction play in connectivity. I sampled three sites within 10 km of each other in the Wellington Region (WR), and another site on an island (Kapiti Island) approximately 50 km north of the WR. At one of the WR sample sites, I monitored a T. burtoni population over two years to examine the dispersal range of asexually reproduced buds and the ability of clones to sexually reproduce. The WR and Kapiti Island populations were strongly genetically differentiated, but within the WR region, two populations were genetically similar, indicative of high connectivity. For the monitored population, asexual bud dispersal was restricted to no greater than 1 m and clonal individuals had reduced sexual reproductive ability. Asexual reproduction did not appear to play an important role in interpopulation connectivity nor gene flow, as buds had low dispersal ability and rarely reproduced. Population structure and connectivity for T. burtoni appear to be largely driven by sexual reproduction, and asexual reproduction instead aids genotype survivorship and population maintenance. These findings highlight that different reproductive modes can differentially contribute to population dynamics in sessile marine organisms, suggesting that predictions about future population viability under changing environments may be difficult to make. In summary, this PhD thesis uses a combination of genetic, histological, field-based and experimental methods to examine species boundaries, reproduction and connectivity for Tethya spp. on rocky reefs of New Zealand. The sympatric nature, complex reproductive ecology and connectivity patterns observed likely shape the complex evolutionary processes occurring in these sponges, including introgressive hybridization and cryptic species. Individuals that showed evidence of possible introgressive events occurred mainly in populations with more restricted gene flow, while the presence of both cryptic species were more prevalent in well connected populations. Such a trend allows for discussion of under what circumstances both of these processes occur. Furthermore, environmental correlates to both sexual and asexual reproduction indicate that both of these modes of reproduction have the potential to be altered with future changes in the environment. As both modes were found to play different roles in gene flow within and between populations, future shifts in climate are also expected to alter population structure and connectivity for these sponges. Such shifts in gene flow will also likely result in changes to species boundaries and thus the overall diversity of this genus. Many other sessile, benthic marine organisms present reproductive traits and behaviours similar to those of Tethya spp., and therefore these results can aid in the interpretation of results for other marine taxa. Overall, this thesis describes the population dynamics of Tethya spp., which are abundant and ecologically important on New Zealand reefs, and provides insight on how temperate sponge populations may fare with climate change, which has important implications for management and conservation efforts.</p>

The distribution of the neutral component of genetic diversity is the interplay of historical and ongoing processes resulting in the species‐specific genetic structure of populations, which can, however, be disrupted by interspecific hybridisation and introgression. In this study, we focused on two species of water frogs, Pelophylax epeiroticus and P. kurtmuelleri, which live in sympatry in the southwestern Balkans, to investigate the rate of hybridisation and population genetic structure using cytogenetic, mitochondrial (ND2) and nuclear DNA (microsatellite) markers. The overall hybridisation rate was 2.6%, with rates reaching up to 10% at specific sites. The course of gametogenesis and the occurrence of later generations of hybrids (beyond the F1 generation) indicate a sexual mode of hybrid reproduction. The bimodal structure of hybrid populations and the rarity of hybrids suggest substantial reproductive isolation between the two species; however, this isolation is unlikely attributable to differences in ecological niche occupation. In P. epeiroticus, sequence variation in the ND2 gene revealed two divergent lineages with a clear geographic pattern that corresponds to the genetic structure in microsatellite markers. In contrast, P. kurtmuelleri populations were not as geographically structured and showed only weak genetic differentiation in both types of markers. Pelophylax epeiroticus was significantly less variable at microsatellite loci compared to P. kurtmuelleri, which, together with the high differentiation of its populations, suggests a stronger influence of genetic drift. We can hypothesise that the differential strength of genetic drift in the two species may lead to unequal interspecific gene flow.

The genetic structure of populations can be both a cause and a consequence of ecological interactions. For parasites, genetic structure may be a consequence of preferences for host species or of mating behaviour. Conversely, genetic structure can influence where conspecific interactions among parasites lay on a spectrum from cooperation to conflict. We used microsatellite loci to characterize the genetic structure of a population of the socially parasitic dulotic (aka "slave-making") ant (Polyergus mexicanus), which is known for its host-specificity and conspecific aggression. First, we assessed whether the pattern of host species use by the parasite has influenced parasite population structure. We found that host species use was correlated with subpopulation structure, but this correlation was imperfect: some subpopulations used one host species nearly exclusively, while others used several. Second, we examined the viscosity of the parasite population by measuring the relatedness of pairs of neighbouring parasitic ant colonies at varying distances from each other. Although natural history observations of local dispersal by queens suggested the potential for viscosity, there was no strong correlation between relatedness and distance between colonies. However, 35% of colonies had a closely related neighbouring colony, indicating that kinship could potentially affect the nature of some interactions between colonies of this social parasite. Our findings confirm that ecological forces like host species selection can shape the genetic structure of parasite populations, and that such genetic structure has the potential to influence parasite-parasite interactions in social parasites via inclusive fitness.

Advances in molecular techniques have enabled the study of genetic diversity and population structure in many different contexts. Studies that assess the genetic structure of cetacean populations often use biopsy samples from free-ranging individuals and tissue samples from stranded animals or individuals that became entangled in fishery or aquaculture equipment. This leads to the question of how representative the location of a stranded or entangled animal is with respect to its natural range, and whether similar results would be obtained when comparing carcass samples with samples from free-ranging individuals in studies of population structure. Here we use tissue samples from carcasses of dolphins that stranded or died as a result of bycatch in South Australia to investigate spatial population structure in two species: coastal bottlenose (Tursiops sp.) and short-beaked common dolphins (Delphinus delphis). We compare these results with those previously obtained from biopsy sampled free-ranging dolphins in the same area to test whether carcass samples yield similar patterns of genetic variability and population structure. Data from dolphin carcasses were gathered using seven microsatellite markers and a fragment of the mitochondrial DNA control region. Analyses based on carcass samples alone failed to detect genetic structure in Tursiops sp., a species previously shown to exhibit restricted dispersal and moderate genetic differentiation across a small spatial scale in this region. However, genetic structure was correctly inferred in D. delphis, a species previously shown to have reduced genetic structure over a similar geographic area. We propose that in the absence of corroborating data, and when population structure is assessed over relatively small spatial scales, the sole use of carcasses may lead to an underestimate of genetic differentiation. This can lead to a failure in identifying management units for conservation. Therefore, this risk should be carefully assessed when planning population genetic studies of cetaceans.

Organisms with sexual and asexual reproductive systems benefit from both types of reproduction. Sexual recombination generates new combinations of alleles, whereas clonality favours the spread of the fittest genotype through the entire population. Therefore, the rate of sexual vs. clonal reproduction has a major influence on the demography and genetic structure of natural populations. We addressed the effect of reproductive system on populations of the dinoflagellate Alexandrium minutum. More specifically, we monitored the spatiotemporal genetic diversity during and between bloom events in two estuaries separated by 150 km for two consecutive years. An analysis of population genetic patterns using microsatellite markers revealed surprisingly high genotypic and genetic diversity. Moreover, there was significant spatial and temporal genetic differentiation during and between bloom events. Our results demonstrate that (i) interannual genetic differentiation can be very high, (ii) estuaries are partially isolated during bloom events and (iii) genetic diversity can change rapidly during a bloom event. This rapid genetic change may reflect selective effects that are nevertheless not strong enough to reduce allelic diversity. Thus, sexual reproduction and/or migration may regularly erase any genetic structure produced within estuaries during a bloom event.