Life History Studies on the Genus Trillium (Liliaceae) III. Reproductive Biology of Six Sessile-flowered Species Occurring in the Southeastern United States with Special Reference to Vegetative Reproduction

Abstract Life history characteristics, e.g., stage class structures of natural populations and associated reproductive characteristics, were studied for six sessile-flowered, diploid Trillium (Liliaceae) species that are restricted to the Coastal Plain region of the southeastern United States.
These sessile-flowered species reproduce primarily by vegetative propagation. Leaf area measurements were used to categorize stage class distributions of natural populations. Populations propagating mainly by vegetative reproduction had non-sigmoidal, convex-shaped population structures with most plants being in the intermediate stage classes. Low numbers of individuals in smaller stage classes and an abundance of intermediate sized individuals are due to the fact that vegetative propagules are the primary method for maintenance of local populations.
Characteristics associated with sexual reproduction were also examined. Seed output averaged about 20 seeds per plant, except for T. lancifolium which produced 9.7 seeds on the average. Seed setting rates varied between 15.61–44.15% and mean seed weight varied from 4.89 to 10.75 mg. The dominance of vegetative versus sexual reproduction may be due to the fact that all six species occur in ecologically unstable flood plain habitats.     

Morphological variation and female reproductive success in two sympatric Trillium species: evidence for phenotypic selection in Trillium erectum and Trillium grandiflorum (Liliaceae)

I investigated the mating systems and phenotypic variation of two sympatric spring ephemerals, Trillium erectum and T. grandiflorum (Liliaceae), and phenotypic selection acting through female reproductive success for 11 morphological characters in five sympatric populations of the two species. I examined the degree of self-compatibility, pollinator-visitation rates, and pollen limitation of fruit and seed production in both species. Both Trillium species were self-compatible, but outcrossed flowers produced more successful fruits and seeds than self-pollinated flowers. Pollinator-visitation rates to the two species were low compared to other insect-pollinated spring ephemerals. In addition, both T. erectum and T. grandiflorum experienced pollen limitation in fruit and/or seed production; however, levels of fecundity in both species may be influenced by resource availability as well. I found significant phenotypic variation in 11 morphological characters within and among the five study populations. The sizes of all morphological characters were positively correlated. In general, larger T. erectum and T. grandiflorum produced more seeds. Phenotypic selection analysis revealed that direct and indirect selection acted on the size of morphological characters for both species. But there was no detectable selection acting on plant shape. This study reveals that variation in plant size exists within and among populations of both species, and this variation is associated with variance in female reproductive success. Spatial and temporal variation in pollinator and/or resource abundance may play a role in the phenotypic variation exhibited by both Trillium species.     

Differentiation Patterns of Reproductive Systems in the Genus Trillium

Abstract The differentiation patterns of reproductive systems, including breeding and pollination systems, and perennation strategies of the genus Trillium were critically studied. The pedicellate- and sessile-flowered species with different floral morphologies and ecological distributions exhibited distinct modes of reproductive systems in relation to their habitat conditions. The pedicellate-flowered species occurring in stable mesic temperate deciduous forests reproduce exclusively by seeds, while sessile-flowered species growing in the alluvial flood plain habitats depend to a large extent on vegetative reproduction to compensate for the insufficient sexual reproduction under ecologically unstable conditions, e.g., frequent disturbance due to flooding. These sessile-flowered species are characterized by low ovule numbers, subsequent low seed outputs and low fecundity levels. In contrast to the sessile-flowered species, the pedicellate-erect-flowered species showed very high seed setting rates of over 50%. Breeding experiments for four Japanese species suggest that, although they substantially possess the capability of both inbreeding and outbreeding, the high fecundity levels are maintained by predominant inbreeding system. Furthermore, resource levels in the stock organs of these species obviously determine fecundity levels. As a consequence the amount of reserved food in the rhizomes transferred to the next season is determined, which gurantees a continuous reproductive activity of a given individual plant.     

Life History Studies on the Genus Trillium (Liliaceae) I. Reproductive Biology of Four Japanese Species

Abstract Four Japanese Trillium (Liliaceae) species which are representative perennial herbs of the temperate deciduous forests— Trillium kamtschaticum (2x), T. apetalon (4x), T. tschonoskii (4x) and T. smallii (6x)—were studied for their reproductive characteristics, e.g., patterns of reproductive resource allocation and reproductive output.
In spite of the differences in ploidy levels (from 2x to 6x), all four species showed very similar reproductive traits. It became evident that in response to the increase in reproductive allocation to total reproductive organs (RA), the number of seeds produced per plant (P_N) clearly increased. This trend is well in accord with the relationship found in four North American species and also several temperate woodland perennial herbs which occur in closed, stable and predictable environments, and possess typical xenogamous breeding systems. Although there is no conspicuous trend between ploidy levels and P_N, one of the significant differences noted in this study was in seed weight, which ranged from 2.93 mg in diploids, to 3.42–3.45 mg in tetraploids, and to 4.47 mg in hexaploids.     

Predicting mating patterns from pollination syndromes: the case of "sapromyiophily" in Tacca chantrieri (Taccaceae)

Tacca, a genus of tropical herbs, possesses near black flowers, conspicuous involucral bracts and whisker-like filiform bracteoles. These features have been assumed to function as a "deceit syndrome" in which reproductive structures resemble decaying organic material attracting flies that facilitate cross-pollination (sapromyiophily). We investigated pollination and mating in Tacca chantrieri populations from SW China to evaluate this assumption. Contrary to this expectation, populations were highly selfing. Pollinator visitation was infrequent and bagged flowers set abundant seed. Pollen loads on stigmas indicated autonomous self-pollination, some of which occurred prior to flower opening. The seed set of inflorescences with bracts and bracteoles removed was not significantly different from unmanipulated inflorescences, suggesting that these structures play a limited role in pollinator attraction, at least at our study sites. Pollen : ovule ratios averaged 49, a value expected in a highly selfing species. Selfing rates estimated in four populations using allozyme markers averaged 0.86 (range 0.76-0.94), thus corroborating this inference. Our results indicate that despite considerable investment in extravagant display, populations of T. chantrieri are highly selfing. We propose several hypotheses to resolve this paradox and argue that future studies of pollination syndromes would benefit by investigation of both pollination and mating biology.     

Impact of tree leaf phenology on growth rates and reproduction in the spring flowering species Trillium erectum (Liliaceae)

We investigated the impact of overstory tree leaf phenology on growth rates, carbon allocation pattern, and fruit characteristics in the spring flowering species, Trillium erectum (Liliaceae). Air temperature, overstory canopy closure, and T. erectum phenology were monitored at three locations following a latitudinal gradient in Québec, Canada. Northern sugar maple trees leaf out at cooler temperatures than more southern populations, while Trillium development was initiated at the same soil temperature irrespective of the latitude. Therefore, in northern areas, the time between initiation of T. erectum leaf expansion and canopy closure was shorter than in southern areas, which left less time for northern plants to accumulate reserves before canopy closure. Differences in growth patterns were noted between T. erectum populations. From a south-north gradient, investment to reproduction, total plant biomass, and annual growth rate decreased, while specific leaf area and stem height increased, indicating shade acclimation. The length of the high light period in early spring seems to be a determinant for spring flowering plants' growth and reproduction and may explain the northern distribution limit of some of these species.     

Reproductive indicators in natural populations of Douglas-fir in Mexico

The reproductive capacity in nine Mexican Douglas-fir populations was determined by analyzing seed production traits from 144 trees collected in 2001. Significant variation was found for all traits among populations; they contributed between 21% and 43% of total phenotypic variation found in these traits, indicating broad differences in reproductive capacity for that particular year. Seed efficiency (filled seed/seed potential) varied from 14% to 42% among populations; all populations from Central Mexico had a seed efficiency below 25%. The proportion of developed that were empty seeds varied from 0.40 to 0.81 among populations, whereas seed size varied also from 0.88 to 1.21 g per 100 seeds among them. Average ratio of filled seed weight to cone weight (reproductive efficiency) was 29.6 mg g⁻¹, but it varied three-fold between populations with extreme values. Populations with larger cones had greater seed potential and heavier seed but not necessarily higher reproductive or seed efficiency. Most reproductive indicators were significantly correlated with latitude, with lower values in the marginal populations from Central Mexico, in the southern extreme of the species range. Given these results, the need for conservation of Douglas-fir populations in Central Mexico is discussed.     

A field study of seed germination in the endangered Trillium reliquum Freeman (Trilliaceae)

Previous studies examining the seeds of most Trillium species have reported double dormancy, a type of seed dormancy where two cold periods and one warm period are needed for complete germination. In the present paper, we describe a field study examining the federally endangered Trillium reliquum Freeman (Trilliaceae) in which moderate to high numbers of seeds germinated after one winter following seed production. Sixteen baskets with seeds were placed in four T. reliquum populations (four baskets in each population) in Georgia, USA, in June 2005. In spring 2006, all seed baskets contained seedlings. Germination percentages ranged from 33.3 to 83.3% across sites with a mean of 56.9 ± 3.9%. Trillium reliquum had higher germination percentages compared with other field‐based germination studies with other Trillium species. Our findings will inform future demographic studies of T. reliquum and suggest that double dormancy in seeds may not be as widespread as previously reported within the genus Trillium.     

Timing of canopy closure influences carbon translocation and seed production of an understorey herb, Trillium apetalon (Trilliaceae)

BACKGROUND AND AIMS: The light availability on a temperate, deciduous-forest floor varies greatly, reflecting the seasonal leaf dynamics of the canopy trees. The growth and/or reproductive activity of understorey plants should be influenced by the length of the high-irradiance period from snowmelt to canopy closure. The aim of the present study was to clarify how spring-blooming species regulate the translocation of photosynthetic products to current reproduction and storage organs during a growing season in accordance with the changing light conditions. METHODS: Growth pattern, net photosynthetic rate, seed production, and shoot and flower production in the next year of Trillium apetalon were compared between natural and experimentally shaded conditions. Furthermore, translocation of current photosynthetic products within plants was assessed by a labelled carbon-chase experiment. KEY RESULTS: During the high-irradiance period, plants showed high photosynthetic ability, in which current products were initially used for shoot growth, then reserved in the rhizome. Carbon translocation to developing fruit occurred after canopy closure, but this was very small due to low photosynthetic rates under the darker conditions. The shading treatment in the early season advanced the time of carbon translocation to fruit, but reduced seed production in the current year and flower production of the next year. CONCLUSIONS: Carbon translocation to the storage organ had priority over seed production under high-irradiance conditions. A shortened bright period due to early canopy closure effectively restricts carbon assimilation, which greatly reduces subsequent reproductive output owing to low photosynthetic products for fruit development and small carbon storage for future reproduction. As populations of this species are maintained by seedling recruitment, acceleration of canopy closure timing may influence the maintenance and dynamics of populations.     

Reproductive attributes of <Emphasis Type="Italic">Campanula</Emphasis> populations from Mt Olympos,Greece

We studied five reproductive attributes (fruit production and volume, seed content and weight, and seed germination) of different populations of Campanula species occurring along the elevation gradient of Mt Olympos, the highest mountain of Greece (2917 m), in order to examine whether and how they change with elevation, and if there are trade-offs among them. We also studied flower and fruit (reproductive) losses and population features in order to estimate whether there might be a threat to the survival of Campanula species on Mt Olympos. The genus is represented by nine species, ranging from cosmopolitan to steno-endemic. All species were found non-autogamous, and hence they depend on pollinators for their sexual reproduction. At the genus level, there is a trade-off between seed content per fruit and seed weight. At the species level, this is manifested only for C. glomerata, which produces the heaviest seeds, far exceeding all other species, and for C. spatulata, represented on Mt Olympos by two elevation distinct subspecies. Changes of the reproductive attributes are not continuous all along the elevation gradient at the genus level nor are they monotonic at the species level. Nevertheless, upland Campanula populations (≥1500 m) produce fewer fruits per individual, containing fewer but heavier seeds than lowland populations. This indicates a transition from ‘r’ to ‘K’ strategies, from low to high elevations, respectively. The patterns of change of reproductive attributes indicate prevalence of phylogenetic over environmental constraints. Laboratory tests showed that for many populations germinability is high at 15 °C, but there are both inter- and intraspecific differences regarding temperature optima. Ecotypic variation among populations of the same species at different elevations was evident; populations from higher elevations had lower optima and vice versa. Reproductive losses, ranging from zero to over 90%, occurred mainly at the stage of fruit maturation; grazing by sheep and goats was the usual cause. Though such losses might affect the short-lived species (the annual C. sparsa and the biennial C. lingulata), which depend entirely on fruit maturation and seed dispersal for their reproduction, there does not seem to be a serious threat to the survival of Campanula species on the mountain.     

Seed viability in declining populations of Caladenia rigida (Orchidaceae): are small populations doomed?

Despite comparatively good rates of pollination and seed production, some populations of the endangered terrestrial orchid Caladenia rigida continue to decline. To determine whether seed quality may be limiting reproductive potential, we assessed seed viability among declining populations of C.?rigida (in the southern part of its distribution) and among populations that are regarded as stable (in the northern part of its distribution). We also compared differences in seed viability to plant traits, population size and habitat characteristics (soil properties, canopy cover, presence of proximate vegetation). Seed capsules from southern populations were significantly smaller, with only 9% of seeds being viable, compared to 36% in capsules from northern populations. Soil phosphorus concentrations differed between regions, but other habitat characteristics did not correlate with seed viability. Using calculations based on seedling recruitment data from other Caladenia species, we predict that seed output is insufficient to ensure the long-term persistence of the smallest C.?rigida populations.     

The evolution of self-compatible and self-incompatible populations in a hermaphroditic perennial, <Emphasis Type="Italic">Trillium camschatcense</Emphasis> (Melanthiaceae)

The evolution of selfing from outcrossing ancestors is known to have occurred repeatedly in angiosperms. Theoretical studies have argued that the transition from outcrossing to selfing is accomplished more easily than the reverse case, and phylogenetic analyses involving self-compatible (SC) and self-incompatible (SI) species has basically supported this assumption. The evolutionary direction of self-compatibility and self-incompatibility was examined in Trillium camschatcense, which contains geographically widespread SC populations, and restricted SI populations. Ecological surveys have revealed that the SC populations were suitable for outcrossing, and selfing in these populations did not confer any fitness advantage. Since reproductive fitness indicates the possibility of an evolutionary shift from self-compatibility to self-incompatibility, the phylogenetic relationships of SI and SC populations of T. camschatcense were investigated based on cpDNA variations and nuclear DNA microsatellite polymorphisms. Although phylogenetic analyses did not provide credible evidence to determine evolutionary direction, the SI populations turned out to be monophyletic with extremely low genetic differentiation. Based on these results, we proposed two possible scenarios for the evolutionary backgrounds of SI and SC populations in T. camschatcense. The plausibility of each scenario was evaluated based on the reproductive and geographical features of the mating systems. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

State of the reproductive system of populations of species of the genus Glycyrrhiza L. (Fabaceae)

Reproductive features of the valuable medicinal herbs Glycyrrhiza glabra L., G. uralensis Fisch., and G. korshinskyi Grig. were investigated; an assessment was made of the state of the reproductive systems of populations located under introduction conditions in the Central Urals and in the zone contact of the species ranges in the southern cis-Urals. The states of the male and female reproductive systems, seed productivity elements, pollination features, and the viability of seeds produced by different pollination treatments were studied. For the three Glycyrrhiza species, the seed productivity indices were found to decrease considerably starting from the early stages of development of the female reproductive system and continuing to seed maturity. A quantitative analysis of seed productivity elements taking into account the flowering duration of the species showed that the seed production system of the species is very efficient; even in the introduced populations located north of the range, it is able to produce viable seeds in years with favorable growing conditions. Qualitative and quantitative indices of the reproductive system were used to characterize samples from the zone of contact between the ranges. Embryological analysis confirmed the possibility of interspecific hybridization and the prevalence of hybrid forms among the samples. A broad distribution of interspecific hybridization in Glycyrrhiza in various parts of the ranges is assumed. Glycyrrhiza species are characterized by seed and vegetative reproduction. Vegetative reproduction is limited in space but efficiently ensures reproduction of individuals and formation of vegetative clones. Seed reproduction is characterized by indices such as amphimictic seed formation, strict entomophily, and interspecific compatibility. Seed reproduction is a factor that controls the possibility of wide propagation of the genetic material in space and, hence, the formation of mixed and hybrid populations leading to a higher intraspecific and interspecific diversity of the genus.     

Restricted seed contribution and clonal dominance in a free-floating aquatic plant <Emphasis Type="Italic">Utricularia australis</Emphasis> R. Br. in southwestern Japan

Utricularia australis R. Br. is an aquatic angiosperm species common in natural and irrigation ponds in temperate regions. This species reproduces both sexually and vegetatively, but in southwestern Japan the occurrence of male-sterile populations, in which plants produce no pollen and propagate only vegetatively, has been recorded. We studied the reproductive contribution of seeds in normal pollen-producing populations using isozyme analyses, a pollination experiment under culture and field observations. Although seedlings obtained from controlled mating indicated segregation of isozyme, polymorphism of the isozyme genotype was detected mainly among populations, but rarely within each pond population. This suggested clonal dominance and rarity of seed or seedling survival in natural populations. In the pollination experiment, the mean seed set ratio in cross-pollination between plants of the same isozyme genotype (7.6%) did not differ significantly from self-pollination (7.6%), but was lower than cross-pollination between plants of different genotypes (45.7%). The low ratio in crossing between the same genotype plants was ascribed to the clonality of the parents. In general, these results corresponded with the low ratios in seed setting observed in natural populations (7.9–13.7%). All the male-sterile populations we surveyed showed the same genotype, thus male sterility in the study area was considered to have the same origin.     

ECOLOGICAL GENETICS OF THE COLONIZING ABILITY OF ROSE CLOVER (TRIFOLIUM HIRTUM ALL.)

Thirteen populations sampled from among the oldest plantings of rose clover (Trifolium hirtum All.), introduced into the California range during the late 1940's, and 12 roadside populations established through natural colonization were compared for their genetic and demographic features. Roadside colonies showed a greater amount of reproductive effort in terms of a larger number of heads per plant and larger calyx, lower rate of seed carryover, higher and more stable plant density, lower seedling survivorship, and earlier flowering. The calyx was more hirsute in roadside collections and remained attached with the seed, a feature accounting for higher germination probabilities on the soil surface or in litter along the roadsides, in contrast to the range where grazing animals would often work the seed into the soil. Outcrossing rates are slightly higher in roadside colonies in which genetic polymorphisms at three marker loci represented as high levels of genetic variability as in the range populations. The colonizing success of rose clover seems to be largely determined by a few and rapid morphological changes and by the retention of some outbreeding and genetic variation. Such joint analyses of genetic and demographic features of colonizing species are needed to support various deductions about the characteristics of “ideal” weeds and colonizers.     

Signals Can Trump Rewards in Attracting Seed-Dispersing Ants

Both rewards and signals are important in mutualisms. In myrmecochory, or seed dispersal by ants, the benefits to plants are relatively well studied, but less is known about why ants pick up and move seeds. We examined seed dispersal by the ant Aphaenogaster rudis of four co-occurring species of plants, and tested whether morphology, chemical signaling, or the nutritional quality of fatty seed appendages called elaiosomes influenced dispersal rates. In removal trials, ants quickly collected diaspores (seeds plus elaiosomes) of Asarum canadense, Trillium grandiflorum, and Sanguinaria canadensis, but largely neglected those of T. erectum. This discrepancy was not explained by differences in the bulk cost-benefit ratio, as assessed by the ratio of seed to elaiosome mass. We also provisioned colonies with diaspores from one of these four plant species or no diaspores as a control. Colonies performed best when fed S. canadensis diaspores, worst when fed T. grandiflorum, and intermediately when fed A. canadense, T. erectum, or no diaspores. Thus, the nutritional rewards in elaiosomes affected colony performance, but did not completely predict seed removal. Instead, high levels of oleic acid in T. grandiflorum elaiosomes may explain why ants disperse these diaspores even though they reduce ant colony performance. We show for the first time that different elaiosome-bearing plants provide rewards of different quality to ant colonies, but also that ants appear unable to accurately assess reward quality when encountering seeds. Instead, we suggest that signals can trump rewards as attractants of ants to seeds.     

Dioecious plants are more precocious than cosexual plants: A comparative study of relative sizes at the onset of sexual reproduction in woody species

The reproductive capacities of dioecious plant species may be limited by severe pollen limitation and narrow seed shadows for the two reasons. First, they are unable to self‐pollinate, and seed production occurs only with pollinator movement from males to females. Second, only 50% of the individuals in populations contribute to seed production. Despite these handicaps, dioecious plants maintain their populations in plant communities with cooccurring cosexual plants, and no substantial difference in population growth rates has been found between dioecious and cosexual plants. Hence, dioecious plants are thought to mitigate these disadvantages by adopting ecological traits, such as insect pollination, animal‐dispersed fleshy fruits, and precocious flowering. We studied the relationship between flowering and plant size in 30 woody species with different sex expressions, leaf habits, fruit types, and maximum plant sizes. The study site was located in an evergreen broad‐leaved forest on the island of Honshu, Japan. A phylogenetic linear regression model showed that dioecious species tended to mature at smaller sizes than did cosexual taxa. At the population level, given equal plant densities and reproductive efforts, the precocity of dioecious plants could serve as one of the factors that mitigate the limitations of pollen and seed‐shadow handicaps by increasing the density of reproductive individuals in the population. At the individual level, smaller size of onset of flowering may play a role in enhancing reproductive success over a lifetime by increasing reproductive opportunities. We discussed the possible effect of the relationship between precocity and some ecological traits of dioecious plants, such as small flowers pollinated by unspecialized insects, fleshy fruit dispersed by animals, and their preferential occurrence in the tropics and in island habitats. The universality of precocity among dioecious plants should be investigated in diverse plant communities. Such studies will increase our understanding of the evolution of plant breeding systems.     

Experimental analyses of the evolutionary processes inTrillium

In the Japanese species ofTrillium examined here, every interspecific cross produced healthy F₁ progeny at the rate of about 50 to 60 per cent of the total inborn ovules except forT. hagae. These figures were not appreciably different from those of self-pollinated or sib-mated examples of these species.Trillium hagae, a putative natural hybrid betweenT. kamtschaticum (2n=10) andT. tschonoskii (2n=20), accepted all foreign sources of pollen grains in manual pollination. These and other crossing experiments suggest that no appreciable barrier has developed that might cause interspecific crossability and that they are habitual inbreeders. Despite theri crossability, a very low rate of actual pollen flow was detected by examining electrophoretic phenotypes of seeds taken from sympatric populations of these species.     

Reproductive and genetic characteristics of rare,disjunct pitch pine populations at the northern limits of its range in Canada

Pitch pine, Pinus rigida Mill., is a rare species in Canada, existing as a disjunct population in the St. Lawrence River Valley in eastern Ontario and two northern outlier stands in southern Quebec along Canada's southern border with the United States. Reproductive and genetic characteristics of these small, scattered stands were investigated to develop a foundation for management and restoration in the event of range expansion northwards under anticipated climate warming. Seed yields and seed quality appear to be comparable to other eastern conifers, and to pitch pine at the center of its geographic range. For seed and seedling growth traits, most of the variation was attributable to differences among trees within stands and, to a lesser extent, among stands within a population; whereas the population effect was non-significant. For reproductive traits, such as numbers of filled and empty seeds per cone, reproductive efficiency, and inbreeding estimates, high levels of variation (ranging from 26% to 33%) were found among stands, suggesting that stand structural features, such as stand size and tree density within stands, play an important role in pollination environment and overall reproductive success. Estimates of genetic diversity at 32 allozyme gene loci indicate that these small, isolated stands have maintained relatively high levels of genetic diversity compared with populations at the center of its geographic range, and also relative to other widely dispersed eastern conifers. The relatively high levels of viable seed production and genetic diversity in native pitch pine populations indicate that native Canadian populations may be suitable seed sources for species restoration and range expansion in Canada.