Development of microsatellite markers for the dwarf bamboo species Sasa cernua and Sasa kurilensis (Poaceae) in northern Japan

Ten polymorphic microsatellite markers were isolated from the dwarf bamboo species Sasa cernua and Sasa kurilensis. The applicability of these markers was confirmed by genotyping of open‐pollinated seeds and leaf samples from natural populations. Genotypes of seeds from each culm shared at least one allele from the maternal parent without contradiction. All 10 loci were polymorphic in S. cernua with 2–15 alleles (average H_E = 0.532). Eight loci were polymorphic in S. kurilensis with 2–10 alleles (average H_E = 0.532). These markers will be useful in detailing the extent of clonal and sexual reproduction in these species.     

VERY LOW GENETIC HETEROZYGOSITIES IN SEXUAL AND AGAMOSPERMOUS POPULATIONS OF EUPATORIUM ALTISSIMUM (ASTERACEAE)

An electrophoretic study revealed very low genetic heterozygosities for both sexual and agamospermous populations of Eupatorium altissimum in eastern North America. Low heterozygosity or gene diversity (H_T = 0.03) can be explained by the small sizes of the extant sexual populations, estimated to be on the order of 10⁴. There was no substantial difference in observed heterozygosity between sexual and agamospermous races. This result shows that a higher level of heterozygosity is not a prerequisite for the evolution of agamospermous races from the sexual ancestor.     

AFLP markers closely linked to a major gene essential for nucellar embryony (apomixis<Emphasis Type="Italic">)</Emphasis> in <Emphasis Type="Italic">Citrus maxima</Emphasis> × <Emphasis Type="Italic">Poncirus trifoliata</Emphasis>

Some citrus varieties express a form of apomixis termed nucellar embryony in which the adventive embryos develop from nucellus tissue surrounding the embryo sac. This trait results in many seeds containing multiple embryos (polyembryony). Inheritance of the frequency of polyembryony was studied in 88 progeny from a cross of Citrus maxima (monoembryonic) × Poncirus trifoliata (polyembryonic). The frequency of polyembryonic seed produced by each progeny was determined by scoring 100–500 seeds for the number of seedlings to emerge from each seed. Two groups of eight individuals from each extreme of the population were chosen for bulked segregant analysis with amplified fragment length polymorphism markers amplified with 256 primer combinations. Candidate markers identified in the bulks as linked to the trait were tested on the 32 individuals used to create the bulks and then on the remaining plants in the population. Five candidate markers tightly linked to polyembryony in P. trifoliata were identified. Specific marker alleles were present in nearly all progeny that produced polyembryonic seed, and alternate alleles were present in nearly all progeny that produced only monoembryonic seed. The region defined by these markers very likely contains a gene that is essential for the production of polyembryonic seeds by apomixis, but also shows segregation distortion. The proportion of polyembryonic seeds varied widely among the hybrid progeny, probably due to other genes. Scoring 119 progeny of a P. trifoliata selfed population for the closely linked markers and the proportion of polyembryonic seeds confirmed close linkage between these markers and polyembryony.     

Genotypic variation in agamospermous Erigeron compositus (Asteraceae)

Few integrative analyses of the structure of agamospermous plant populations have been conducted. Erigeron compositus occurs in montane western North America and comprises both sexual and agamospermous populations. Sexual E. compositus has previously been characterized as outcrossing and predominantly diploid (2n = 18). Agamic E. compositus is usually hexaploid (2n = 54), though counts herein range from 2n = 36 to 2n = 80. Starch-gel electrophoresis, cytology, and analysis of pollen production were used to evaluate variation within and among agamospermous populations. Fifteen enzyme loci were used to identify 24 unique multilocus genotypes in seven populations, an average of 3.4 genotypes per population. Proportion of distinct genotypes per population sample size (GIN) and measures of genetic diversity (D) and evenness (E) are 0.10, 0.48, and 0.61, respectively, which indicate that E. compositus maintains levels of diversity similar to other agamospermous taxa. Most agamospermous populations are mosaics comprising groups of genetically distinct individuals that are frequently distinguished by cytotype and capacity for pollen production. The geographical and ecological separation of sexual and agamospermous populations make it unlikely that gene flow from sexual populations is a direct source of genetic variation in agamospermous populations. Instead, crossing between genetically distinct facultative agamosperms probably accounts for most variation. Genetic and morphological evidence document one such putative crossing event. Agamospermous E. compositus is very similar genetically to sexual E. compositus. Allozyme analysis further shows that genetic variation in agamospermous populations is partitioned among a few highly heterozygous genotypes, whereas sexual populations maintain numerous genotypes of relatively low heterozygosity.     

Mating systems in two species ofPotentilla from Alaska

Aspects of reproductive ecology have been studied in two species ofPotentilla (Rosaceae) from interior Alaska. Data were collected to determine mating system traits such as seed: ovule and pollen: ovule ratios, and experiments were carried out in the field in order to study the effect of caging, emasculation, and manual self-pollination on seed set and seed quality. Mating system strategies differ between the two species in the populations examined. High pollen: ovule ratio, high seed: ovule ratio, good autodeposition ability, and unpredictable pollination in concert with self-incompatibility indicate thatP. hookeriana in the population studied has adopted a more or less pseudogamous mating system. In contrast, the data suggest thatP. uniflora in the studied population is approaching a sexual, facultatively outbreeding species. Because both species exhibit ploidy variation, a trait common among agamospermous plants, it is concluded that both species are likely to be facultatively agamospermous but that sexual reproduction is much more important in the population ofP. uniflora than it is inP. hookeriana.     

A taxometric analysis ofLimonium (Plumbaginaceae) in Western Europe

A taxometric analysis ofLimonium species in Western Europe has provided evidence for the subgeneric groupings proposed by other authors. A cluster analysis suggests that the origin of sexual and agamospermous species has taken place separately. Sexual and agamospermous species are usually clustered separately. The lack of a sexual process in many agamospermous species, as indicated by the distribution of incompatibility morphs and by very low pollen fertility, suggests that the evolution of sexual and agamospermous taxa has been by different processes. It is suggested that following the origin of agamospermous species or species groups by hybridisation, subsequent evolution in the agamospermous taxa has occurred by asexual means. Evidence from the distribution of pollen/stigma incompatibility morphs, especially the existence of self-compatible agamospermous species supports this hypothesis.     

Polyembryony and apomixis in Eriotheca pubescens (Malvaceae - Bombacoideae)

Apomixis and adventitious polyembryony have been reported for several species of Bombacoideae, including Eriotheca pubescens, a tree species of the Neotropical savanna (Cerrado) areas in Brazil. However, the origin of polyembryonic seeds and their importance for the reproduction of the species remained to be shown. Here, we analyzed the early embryology of this species to establish the apomictic origin of extranumerary embryos. We also observed the geographic distribution of polyembryony in E. pubescens, and tested if apomixis was related to the source of pollen (self or cross) and population density. Moreover, we tested if polyembryonic apomictic embryos would develop normally into seedlings. In the observed seed primordia, after a relatively long quiescent period, the zygote developed into a sexual embryo concurrently with adventitious apomictic embryos which developed from nucellus cells. Adventitious embryos develop faster than sexual ones and are morphologically similar, so that 44 days after anthesis it was virtually impossible to distinguish and trace the fate of the sexual embryo. Polyembryony is widely distributed in populations some 400 km distant, and only one strictly monoembryonic individual was observed during the study. The number of embryos per seed varied between fruits and individuals but was significantly higher in seeds from cross-pollinations than from selfs, although fruit and seed set after crosses were much lower than after selfs. Embryo development into seedlings depended on their weight at germination, but polyembryonic seeds germinated and produced up to seven seedlings per seed in greenhouse conditions. Adventitious embryony and apomictic seedlings would explain the mostly clonal populations suggested by molecular studies.     

Stomatal size,ploidy and polyembryony in Eriotheca Stellate Trichome Species Complex (Bombacoideae – Malvaceae) in the Cerrados of Brazil

• Geographic parthenogenesis, range expansion of apomictic plants after climate changes, has been described for Northern Hemisphere gametophytic apomicts. But similar trends have been observed for sporophytic apomicts of Cerrado, the savannas in Brazil. Eriotheca pubescens is a common Cerrado tree, an agamic complex of either hexaploid/polyembryonic apomicts or tetraploid/monoembryonic sexual individuals. Some populations have been described as a new species, Eriotheca estevesiae, all included in the Eriotheca Stellate Trichome Species Complex (ESTSC). Since breeding systems and ploidy are clearly associated with polyembryony and stomatal size, we used these ancillary features to map the reproductive and ploidy level traits of E. pubescens and E. estevesiae.
• Leaves and seeds were collected from individuals of 19 populations. Seeds were evaluated for the presence of polyembryony and leaves for stomatal measurements.
• Eight populations were monoembryonic while another eight were polyembryonic and for other three, the embryonic pattern was not readily verified. E. pubescens polyembryonic and hexaploid populations formed a homogeneous group, but monoembryonic plants were more variable. E. estevesiae populations were monoembryonic with smaller stomata. In contrast, some E. pubescens monoembryonic populations further south presented larger stomata. Despite these outliers, possibly mixed populations, stomatal size and embryonic pattern differed from northern to southern populations.
• Embryonic pattern and stomatal size indicated that northernmost populations of Eriotheca STSC (E. estevesiae) are diploid and sexual. Southernmost populations, mostly polyembryonic and with large stomata, are hexaploid and apomictic. This is in agreement with geographic parthenogenesis and range expansion of apomictic lineages to southern habitats available after the last glacial maximum.

     

Population genetic structure in European populations of Spiranthes romanzoffiana set in the context of other genetic studies on orchids

Spiranthes romanzoffiana Cham. is restricted in Europe to the British Isles, where it is recognised as a conservation priority species due to frequent extirpation of populations along with no evidence of seed set; vegetative reproduction has been invoked as the sole means of perpetuation and dispersal. To investigate the reproductive ecology of this species, 17 populations have been sampled for chloroplast microsatellites and amplified fragment length polymorphisms (AFLPs). These markers revealed a previously unsuspected genetic-geographic split in the species, which correlates with differences in patterns of within-population variation. Northern populations were fixed for one chloroplast haplotype but showed high levels of AFLP genotypic diversity consistent with sexual reproduction (proportion of genotypes distinguishable, PD = 0.98). More southerly populations showed fixed differences from the northern populations in their chloroplast haplotype and for 10 AFLP markers. They harboured only 12 unique multilocus genotypes among 113 individuals from six populations (PD = 0.11). These genotypes differed mostly by single bands, and none by more than 4/138 loci, with identical multilocus genotypes occurring in widely separated populations. This uniformity in southern populations is consistent with agamospermous or autogamous reproduction, and/or an extreme population bottleneck. Finally, the observed patterns of population differentiation in S. romanzoffiana are compared with other studies of orchids, revealing a wide range of values that belie recent contrasting published generalisations that claim that orchids show either higher, or lower, levels of population differentiation than other plant families.     

Occurrence of Colombian datura virus in the Terrestrial Orchid, Spiranthes cernua

Colombian datura virus was detected in the terrestrial orchid Spiranthes cernua using a combination of enzyme‐linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). The PCR strategy that we used to detect and confirm the identity of this virus has subsequently been used as a generalized procedure to confirm the identity of other potyviruses in ornamental species. This is the first report of this virus in an orchid and only the second report of a virus in this species.     

Formation of a hybrid triploid agamosperm on a sexual diploid plant: evidence from progeny tests in Taraxacum platycarpum Dahlst.

Agamospermous dandelions of hybrid origin between a native sexual diploid species (Taraxacum platycarpum Dahlst. or T. japonicum Koidz.) and an alien agamospermous triploid [T. officinale Weber and T. laevigatum (Willd.) DC.] are now widely distributed throughout mainland Japan. These hybrid dandelions are known to be genetically variable. We hypothesized that this variability is maintained by repeated ongoing hybridization, based on the fact that triploid dandelions not only produce seeds agamospermously, but also produce some functional pollen grains that are able to sire seeds of sexual dandelions. To test this hypothesis, we examined whether heads of Japanese diploid dandelions produce new hybrid seeds after fertilization by pollen from triploid agamosperms under field conditions. One of the 430 tested plants grown from sexual dandelion seeds had morphological and molecular characteristics, which are consistent with a hybrid origin. The plant formed a hybrid surrounded by many individuals having recurved involucral bracts, in which frequency of T. officinale was very low (3.5 %). Cytological data and bagging experiments demonstrate triploidy and asexual seed production of the hybrid. Taken together, these results supported that the new hybrid is probably derived from a backcross of a hybrid to the native sexual species. Our findings provide evidence for the evolution of a new agamosperm through interspecific hybridization as a contemporary population process.     

Optimum reproduction and dispersal strategies of a clonal plant in a metapopulation: a simulation study with Hieracium pilosella

Clonal spread is favoured in many plants at the expense of seed production in order to expand rapidly into open habitats or to occupy space by forming dense patches. However, for the dynamics of a population in a patchy landscape seed dispersal remains important even for clonal plants. We used a spatially explicit individual-based metapopulation model to examine the consequences of two trade-offs in Hieracium pilosella L: first, between vegetative and sexual reproduction, and second, between short and far-distance dispersal of seeds. Our main question was, what are the environmental conditions that cause a mixed strategy of vegetative and sexual reproduction to be optimal. The model was parameterised with field data on local population dynamics of H. pilosella. Patch dynamics were given firstly by disturbance events that opened patches in a matrix of a clonal grass that were colonisable for H. pilosella, and secondly by the gradual disappearance of H. pilosella patches due to the expanding grass. Simulations revealed opposing selection pressures on traits determined by the two trade-offs. Vegetative reproduction is favoured by local dynamics, i.e. the need for maintenance and expansion of established populations, whereas seed production is favoured by the necessity to colonise empty habitats. Similar pressures act on the proportion of seeds dispersed over short and far distances. Optimum reproductive and dispersal strategies depended on habitat quality (determined by seedling establishment probability), the fraction of dispersed seeds, and the fraction of seeds lost on unsuitable ground. Under habitat conditions supporting moderate to low seedling establishment, between 20% and 40% of reproductive effort in H. pilosella should be devoted to sexual reproduction with at least 10% of the seeds dispersed over distances suitable to attain empty patches. We conclude that in a spatially heterogeneous landscape sexual seed production in a clonal plant is advantageous even at the expense of local vegetative growth.     

Plant clonality, mutation, diplontic selection and mutational meltdown

Apomixis is a very common characteristic in vascular plants. It occurs in two general forms: either subversion of the sexual system (agamospermous seeds or apogamous sporophytes in non-seed plants) or vegetative reproduction. In this communication, only the mutational consequences of vegetative reproduction are considered. Vegetative reproduction involves the replication of apical meristems, especially shoot apical meristems. Three general types of shoot apical meristems occur in the vascular plants: single tetrahedral apical initial, unstratified with impermanent initials and stratified with impermanent initials. Each meristem type has different consequences with regard to mutation, diplontic selection and the possibility of mutational meltdown.  © 2003 The Linnean Society of London. Biological Journal of the Linnean Society , 2003, 79 , 61–67.     

EVOLUTION OF ASEXUALITY VIA DIFFERENT MECHANISMS IN GRASS THRIPS (THYSANOPTERA: Aptinothrips)

Asexual lineages can derive from sexual ancestors via different mechanisms and at variable rates, which affects the diversity of the asexual population and thereby its ecological success. We investigated the variation and evolution of reproductive systems in Aptinothrips, a genus of grass thrips comprising four species. Extensive population surveys and breeding experiments indicated sexual reproduction in A. elegans, asexuality in A. stylifer and A. karnyi, and both sexual and asexual lineages in A. rufus. Asexuality in A. stylifer and A. rufus coincides with a worldwide distribution, with sexual A. rufus lineages confined to a limited area. Inference of molecular phylogenies and antibiotic treatment revealed different causes of asexuality in different species. Asexuality in A. stylifer and A. karnyi has most likely genetic causes, while it is induced by endosymbionts in A. rufus. Endosymbiont‐community characterization revealed presence of Wolbachia, and lack of other bacteria known to manipulate host reproduction. However, only 69% asexual A. rufus females are Wolbachia‐infected, indicating that either an undescribed endosymbiont causes asexuality in this species or that Wolbachia was lost in several lineages that remained asexual. These results open new perspectives for studies on the maintenance of mixed sexual and asexual reproduction in natural populations.     

Reproductive biology of Clintonia udensis

To clarify the life history of Clintonia udensis, we investigated its reproductive systems and spatio‐temporal population structure. Pollination experiments and the observation of floral visitors revealed that C. udensis was compatible with both self‐ and outcross‐pollen, and it potentially produces seeds by insect‐mediated outcrossing in natural conditions. In addition, propagation by clonal reproduction from rhizomes was evident. In this study, it was clarified that C. udensis potentially propagates by sexual and asexual reproduction and maintains its population through a stable frequency of flowering. The differences in the dependence on each reproduction mode could be one of the contributing factors for creating a variety of population sizes and distribution patterns of ramets in populations.     

Demographic consequences of greater clonal than sexual reproduction in Dicentra canadensis

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.     

Habitat‐specific demography,source‐sink dynamics,and the niche of a common shrub in a heterogeneous and fluctuating environment

The occurrence of a species in habitats of varying quality connected through migration can only be understood by detailed investigation of itsdemography. In the Chihuahuan Desert, the common shrub Flourensia cernua is found in both productive and unproductive areas. In the former, both growing and senescent populations are regularly found, while in the latter a low density scattered population persists indefinitely. While precipitation (and its annual stochastic variation) is the same in both habitats, their geomorphological differences produce a sharp difference in the availability of the limiting resource, water. This produces different population dynamics in F. cernua, but also radically different plant communities. Counterintuitively, the low‐resource habitat (LR) supports a scattered, slightly increasing or stable population that coexists with its neighbors and acts as exporter of seeds (source population). In contrast, the high‐resource habitat (HR) allows sporadic recruitment of locally dense patches that tend towards extinction (sink population). The latter is accounted for by the increasing dominance of the grass Pleuraphys mutica. The different dynamics and regulatory mechanisms in each habitat allow the species to occupy a wider distribution than it would have in their absence. The higher abundance of F. cernua in the sink habitat, together with its consequences on community composition and dynamics, questions the idea proposed in the literature that a sink population lives outside its fundamental niche. The study provides support to the notion that the ecological niche of a species cannot be completely characterized by its requirements (e.g. as they relate to physiology), but must also include the complex demographic responses to a spatially and temporally variable environment, which may often include substandard conditions. For the niche concept to retain its usefulness, it must incorporate the demographic response of populations to spatially and temporallyvariable resource supply.     

Mating system,population growth,and management scenario for Kalanchoe pinnata in an invaded seasonally dry tropical forest

Ecological invasions are a major issue worldwide, where successful invasion depends on traits that facilitate dispersion, establishment, and population growth. The nonnative succulent plant Kalanchoe pinnata, reported as invasive in some countries, is widespread in remnants of seasonally dry tropical forest on a volcanic outcrop with high conservation value in east‐central Mexico where we assessed its mating system and demographic growth and identified management strategies. To understand its local mating system, we conducted hand‐pollination treatments, germination, and survival experiments. Based on the experimental data, we constructed a life‐stage population matrix, identified the key traits for population growth, weighted the contributions of vegetative and sexual reproduction, and evaluated management scenarios. Hand‐pollination treatments had slight effects on fruit and seed setting, as well as on germination. With natural pollination treatment, the successful germination of seeds from only 2/39 fruit suggests occasional effective natural cross‐pollination. The ratios of the metrics for self‐ and cross‐pollinated flowers suggest that K. pinnata is partially self‐compatible. Most of the pollinated flowers developed into fruit, but the seed germination and seedling survival rates were low. Thus, vegetative propagation and juvenile survival are the main drivers of population growth. Simulations of a virtual K. pinnata population suggest that an intense and sustained weeding campaign will reduce the population within at least 10 years. Synthesis and applications. The study population is partially self‐compatible, but sexual reproduction by K. pinnata is limited at the study site, and population growth is supported by vegetative propagation and juvenile survival. Demographic modeling provides key insights and realistic forecasts on invasion process and therefore is useful to design management strategies.     

Evidence for multiple, autoploid origins of agamospermous populations in Eupatorium sessilifolium (Asteraceae)

Comparative analyses were made of agamospermous populations of Eupatorium sessilifolium, which have previously been documented to be polyploid, to determine whether they are alloploid or autoploid in origin and to assess the possibility that they have arisen more than once. There was no variability in ITS sequences among seven agamospermous and eight sexual diploid populations of E. sessilifolium, which is consistent with morphological observations in suggesting that the agamospermous populations were autoploids. The ITS sequence characteristic of E. sessilifolium differs from all other North American species by a minimum of 15 changes, and heterogeneity or polymorphism would be expected if the agamospermous populations were alloploids. Analysis of the chloroplast-based trnC-psbM spacer region showed variability among both sexual diploid and agamospermous populations of E. sessilifolium, which suggested that the agamospermous populations stem from multiple origins. Analysis of ISSR data revealed considerable intraspecific variability within E. sessilifolium, and the distribution of variability, with agamospermous populations showing variability from one another, added further evidence for multiple origins of agamospermous populations. The results in conjunction with distributional evidence that the sexual diploid populations of E. sessilifolium are geographically restricted and uncommon suggest that monitoring of populations might be warranted to evaluate whether measures are needed to enhance their continued survival.