Modeling temporal dynamics of genetic diversity in stage-structured plant populations with reference to demographic genetic structure.

Predicting temporal dynamics of genetic diversity is important for assessing long-term population persistence. In stage-structured populations, especially in perennial plant species, genetic diversity is often compared among life history stages, such as seedlings, juveniles, and flowerings, using neutral genetic markers. The comparison among stages is sometimes referred to as demographic genetic structure, which has been regarded as a proxy of potential genetic changes because individuals in mature stages will die and be replaced by those in more immature stages over the course of time. However, due to the lack of theoretical examination, the basic property of the stage-wise genetic diversity remained unclear. We developed a matrix model which was made up of difference equations of the probability of non-identical-by-descent of each life history stage at a neutral locus to describe the dynamics and the inter-stage differences of genetic diversity in stage-structured plant populations. Based on the model, we formulated demographic genetic structure as well as the annual change rate of the probability of non-identical-by-descent (denoted as η). We checked if theoretical expectations on demographic genetic structure and η obtained from our model agreed with computational results of stochastic simulation using randomly generated 3,000 life histories. We then examined the relationships of demographic genetic structure with effective population size Ne, which is the determinants of diversity loss per generation time. Theoretical expectations on η and demographic genetic structure fitted well to the results of stochastic simulation, supporting the validity of our model. Demographic genetic structure varied independently of Ne and η, while having a strong correlation with stable stage distribution: genetic diversity was lower in stages with fewer individuals. Our results indicate that demographic genetic structure strongly reflects stable stage distribution, rather than temporal genetic dynamics, and that inferring future genetic diversity solely from demographic genetic structure would be misleading. Instead of demographic genetic structure, we propose η as an useful tool to predict genetic diversity at the same time scale as population dynamics (i.e., per year), facilitating evaluation on population viability from a genetic point of view.

Temporal skewness of pollination success in the spring ephemeral Trillium camschatcense.

Phenological overlap with pollinators is crucial for reproductive success in insect-pollinated plants. In this study, we examined whether pollinator visitation successfully occurred during an entire flowering season in two populations of the insect-pollinated spring ephemeral Trillium camschatcense in the Tokachi region of Hokkaido, northern Japan. We bagged flowers and excluded pollinator visitation during either the first or the last half of the entire flowering season to compare pollination success between the two periods. The two populations have experienced differing levels of climate warming in the last 60 years, which impacted pollinator visitation. In the population experiencing temperature rise more rapidly, fertilization rate and seed set decreased sharply when bagged during the first half period, indicating that pollinator visitation is skewed to the early part of the flowering season. The temporal skewness of pollination success would be an early warning signal of the impacts of climate warming on the reproductive success of T. camschatcense.

Seasonal variation of responses to herbivory and volatile communication in sagebrush (Artemisia tridentata) (Asteraceae).

Plants can respond to insect herbivory in various ways to avoid reductions in fitness. However, the effect of herbivory on plant performance can vary depending on the seasonal timing of herbivory. We investigated the effects of the seasonal timing of herbivory on the performance of sagebrush (Artemisia tridentata). Sagebrush is known to induce systemic resistance by receiving volatiles emitted from clipped leaves of the same or neighboring plants, which is called volatile communication. Resistance to leaf herbivory is known to be induced most effectively after volatile communication in spring. We experimentally clipped 25 % of leaves of sagebrush in May when leaves were expanding, or in July when inflorescences were forming. We measured the growth and flower production of clipped plants and neighboring plants which were exposed to volatiles emitted from clipped plants. The treatment conducted in spring reduced the growth of clipped plants. This suggests that early season leaf herbivory is detrimental because it reduces the opportunities for resource acquisition after herbivory, resulting in strong induction of resistance in leaves. On the other hand, the late season treatment increased flower production in plants exposed to volatiles, which was caused mainly by the increase in the number of inflorescences. Because the late season treatment occurred when sagebrush produces inflorescences, sagebrush may respond to late herbivory by increasing compensation ability and/or resistance in inflorescences rather than in leaves. Our results suggest that sagebrush can change responses to herbivory and subsequent volatile communication seasonally and that the seasonal variation in responses may reduce the cost of induced resistance.

Demographic response of plant populations to habitat fragmentation and temporal environmental variability.

Many plant species currently exist in fragmented populations of different sizes, while they also experience unpredictable climatic fluctuation over time. However, we still understand little about how plant demography responds to such spatial and temporal environmental variability. We studied population dynamics of an understory perennial herb Trillium camschatcense in the Tokachi plain of Hokkaido, Japan, where a significant effect of forest fragmentation on seedling recruitment was previously reported. Four populations across a range of fragment sizes were studied for 6 years, and the data were analyzed using matrix population models. Per capita fecundity (the number of recruits per plant) varied greatly among populations, but the variation in population growth rates (lambda) was mainly driven by the variation in stasis and growth rates, suggesting that the general trend of reduced fecundity in fragmented populations may not be readily translated into subsequent dynamics. Temporal variation in lambda among years was more than 2 times larger than spatial variation among populations, and this result was likely attributable to the contrasting response of correlation structures among demographic rates. The among-population variation in lambda was dampened by negative covariation between matrix elements possibly due to density-dependent regulation as well as an inherent constraint that some elements are not independent, whereas positive covariation between matrix elements resulted in large temporal variation in lambda. Our results show that population dynamics responded differently to habitat fragmentation and temporal variability of the environment, emphasizing the need to discriminate these spatial and temporal variations in demographic models. Although no populations were projected to be declining in stochastic simulations, correlation between current habitat size and plant density implies historical lambda is positively related to habitat size.

The evolution of self-compatible and self-incompatible populations in a hermaphroditic perennial, Trillium camschatcense (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.

Dynamics of distribution and performance of ramets constructing genets: a demographic-genetic study in a clonal plant, Convallaria keiskei.

BACKGROUND AND AIMS: In clonal plants producing vegetative offspring, performance at the genet level as well as at the ramet level should be investigated in order to understand the entire picture of the population dynamics and the life history characteristics. In this study, demography, including reproduction and survival, the growth patterns and the spatial distributions of ramets within genets of the clonal herb Convallaria keiskei were explored. METHODS: Vegetative growth, flowering and survival of shoots whose genets were identified using microsatellite markers were monitored in four study plots for 3 years (2003-2005). The size structures of ramets in genets and their temporal shifts were then analysed. Their spatial distributions were also examined. KEY RESULTS: During the census, 274 and 149 ramets were mapped in two 1 x 2 m plots, and 83 and 94 ramets in two 2 x 2 m quadrats. Thirty-eight genotypes were identified from 580 samples. Each plot included 5-18 genets, and most ramets belonged to the predominant genet(s) in each plot. Shoots foliated yearly for several years, but flowering ramets did not have an inflorescence the next year. A considerable number of new clonal offspring persistently appeared, forming a bell-shaped curve of the size structure of ramets in each genet. Comparing the structures modelled by the normal distributions suggested variation among ramets belonging to a single genet and variation among genets. Furthermore, spatial analyses revealed clumped and distant distributions of ramet pairs in a genet, in which the distant patterns corresponded to the linearly elongating clonal growth pattern of this species. CONCLUSION: Characteristics of ramet performances such as flowering and recruitment of clonal offspring, in addition to growth, played a large part in the regulation of genet dynamics and distribution, which were different among the studied genets. These might be characteristics particularly relevant to clonal life histories.

Reproductive demography of ramets and genets in a rhizomatous clonal plant Convallaria keiskei.

Clonal growth occurring below the ground makes it difficult to identify individuals and demonstrate the demographic features of a focal plant species. In this study, genotypically identified ramets of a rhizomatous clonal herb, Convallaria keiskei Miq., were monitored for their growth, survival, and reproduction from 2003 to 2006. After the monitoring period, their subterranean organs were excavated to explore the underground connections of established ramets and the direction of clonal growth. We then combined data on the fate of the monitored ramets with the information of rhizome connections, clarifying reproductive demography at both the ramet and genet levels. Although each ramet initiated both sexual reproduction (via flowering) and clonal growth, clonal growth tended to precede sexual reproduction. In a surveyed genet, 51.0% of ramets produced flowers and 29.6% generated clonal offspring during the study period. Consequently, we clarified the reproductive demography of C. keiskei: clonal growth tended to precede flowering in a ramet, and a genet can keep reproducing every season at the genet level, despite a ramet not having inflorescence every year.

Adaptive significance of self-fertilization in a hermaphroditic perennial, Trillium camschatcense (Melanthiaceae).

The evolution of self-fertilization from primarily outcrossing ancestors is one of the most common evolutionary transitions in plants; however, the ecological mechanisms that maintain self-fertilization have remained controversial. Theoretical studies suggest that selfing is advantageous over outcrossing in terms of genetic transmission and assurance of seed production under pollen-limited circumstances. Trillium camschatcense is a herbaceous perennial distributed in Hokkaido and northern Honshu, Japan. Geographical variation in the breeding system (self-compatible, SC; or self-incompatible, SI) has been reported in populations in Hokkaido. Here, we used several SC and SI populations of T. camschatcense to investigate the adaptive significance and the evolutionary basis of self-fertilization. Pollination experiments and genetic analyses demonstrated that the potential availability of outcross pollen in SC populations was sufficient and that the number of pollen donors was equal to that of SI populations. However, despite the high availability of outcross pollen, the SC populations produced seeds predominantly by selfing and so underwent severe inbreeding depression. Although none of the suggested advantages for self-fertilization were supported by our analyses, we propose two possible scenarios for the evolution of self-fertilization in T. camschatcense.

Fine-scale spatial genetic structure within continuous and fragmented populations of Trillium camschatcense.

Spatial genetic structure (SGS) within populations was analyzed for the ling-lived understory perennial herb Trillium camschatcense using allozyme loci. We used Sp statistics to compare SGS between 2 life-history stages, juveniles (J) and reproductives (R), as well as between 2 populations, continuous and fragmented, with different habitat conditions. In the continuous population, significant SGS was detected in both stages but the extent was greatly reduced with the progress of the stage (J, Sp = 0.0475; R, Sp = 0.0053). We inferred that limited seed dispersal and subsequent random loss of individuals from the family patches are responsible for the J and R stage structures, respectively. The fragmented population differed in the patterns of SGS; significant structure was detected in the R stage, but not in the J stage (J, Sp = 0.0021; R, Sp = 0.0165) despite significant positive inbreeding coefficients (J, F(IS) = 0.251). The observed differences in the J-stage structures between populations may be explained by habitat fragmentation effects because reduced recruitment in the fragmented population prevents the development of maternal sibling cohort. Such comparative analysis between populations and life-history stages can be useful to understand the different underlying causes of SGS.

Genetic structure in aquatic bladderworts: clonal propagation and hybrid perpetuation.

BACKGROUND AND AIMS: The free-floating aquatic bladderwort Utricularia australis f. australis is a sterile F(1) hybrid of U. australis f. tenuicaulis and U. macrorhiza. However, co-existence of the hybrids and parental species has not been observed. In the present study, the following questions are addressed. (a) Does the capacity of the two parental species to reproduce sexually contribute to higher genotypic diversity than that of sterile F(1) hybrid? (b) Are there any populations where two parental species and their hybrid co-exist? (c) If not, where and how do hybrids originate? METHODS: The presence and absence of Utricularia was thoroughly investigated in two regions in Japan. An amplified fragment length polymorphism (AFLP) analysis was conducted for 397 individuals collected from all populations (33 in total) where Utricularia was observed. KEY RESULTS: The mean number of genotypes per population (G) and genotypic diversity (D) were extremely low irrespective of the capacity to reproduce sexually: G was 1.1-1.2 and D was 0.02-0.04. The hybrid rarely co-existed with either parental species, and the co-existence of two parental species was not observed. Several AFLP bands observed in the hybrid are absent in both parental genotypes, and parent and hybrid genotypes in the same region do not show greater genetic similarity than those in distant regions. CONCLUSIONS: The capacity to reproduce sexually in parental species plays no role in increasing genotypic diversity within populations. The observed genotypes of the hybrid could not have originated from hybridization between the extant parental genotypes within the study regions. Considering the distribution ranges of three investigated taxa, it is clear that the hybrid originated in the past, and hybrid populations have been maintained exclusively by clonal propagation, which may be ensured by both hybrid vigor and long-distance dispersal of clonal offspring.

Predominance of clonal reproduction, but recombinant origins of new genotypes in the free-floating aquatic bladderwort Utricularia australis f. tenuicaulis (Lentibulariaceae).

Aquatic plants are a biological group sharing several adaptations to aquatic conditions. The most striking evolutionary convergence in this group is the extensive reliance on clonal reproduction, which largely determines the patterns and process of evolution in aquatic plants. Utricularia australis f. tenuicaulis is a free-floating aquatic bladderwort that reproduces both sexually via seeds and clonally via turions and shoot fragments. Amplified fragment length polymorphism analysis was conducted on 267 ramets collected from 30 populations in Japan. The genotypic diversity within populations was extremely low, regardless of the geographical distribution range: the mean number of genotypes per population (G) was 1.4 and the mean genotypic diversity (D), including monoclonal populations, was 0.17. In contrast to the predominance of a few clones within populations, many of the populations investigated had different genotypes; a large portion of the genetic variation was explained by variation among populations. Character compatibility analysis clearly revealed that somatic mutations did not contribute to the origin of genotypic diversity in this aquatic bladderwort; instead, rare-to-sporadic sexual reproduction probably generated new genotypes. Thus, future studies should examine the role of sexual reproduction in this species from the viewpoint of long-term evolutionary benefits.

Hybrid origins and F1 dominance in the free-floating, sterile bladderwort, Utricularia australis f. australis (Lentibulariaceae).

Abandonment of sexual reproduction is a well-known characteristic in aquatic plants, while the causes, levels, and consequences of sterility are often unknown. Utricularia australis f. australis (Lentibulariaceae) is a free-floating, sterile bladderwort distributed widely in temperate and tropical regions. Experimental crosses in cultivated conditions, AFLP analysis, and cpDNA haplotypes of natural populations clearly demonstrated that U. australis f. australis originates from the asymmetric hybridization between two parental taxa: U. australis f. tenuicaulis (mostly as female) and U. macrorhiza (mostly as male). No post-F(1) hybrids were detected using the additive patterns of AFLP bands combined with the observation of extensive sterility in U. australis f. australis. Recurrent hybridizations and subsequent perpetuation by asexual reproduction were demonstrated by the unique, but monomorphic, AFLP genotypes observed in each U. australis f. australis population. Hybrids and parental species did not coexist, implying the superiority of the hybrid U. australis f. australis in certain environmental conditions. It remains unclear whether populations of U. australis f. australis are maintained by colonizing propagules or as relicts of past hybridization events.

Genetic diversity and multilocus genetic structure in the relictual endemic herb Japonolirion osense (Petrosaviaceae).

Plant clonality may greatly reduce effective population size and influence management strategies of rare and endangered species. We examined genetic diversity and the extent of clonality in four populations of the monotypic herbaceous perennial Japonolirion osense, which is one of the most rare flowering plants in Japan. Allozyme analysis revealed moderate levels of genetic variation, and the proportion of polymorphic loci (P=66.7%) was higher than the value for species with similar life-history traits. With four polymorphic loci, 19 multilocus genotypes were observed among 433 aerial shoot samples and 10 (52%) were found only in single populations. The proportion of distinguishable genotypes (PD=0.10) and Simpson's index of diversity (D=0.52) also exhibited moderate levels of genotypic diversity compared to other clonal plants, with genotype frequencies at Hardy-Weinberg equilibrium. The distributions of genotypes were often localized and they were mostly found within a radius of 5 m. Spatial autocorrelation analysis showed that shoot samples located 4 m apart were expected to be genetically independent. The results suggest that the spatial extent of genets was relatively narrow and thus the clonality was not extensive.

Effects of ants, ground beetles and the seed-fall patterns on myrmecochory of Erythronium japonicum Decne. (Liliaceae).

Erythronium japonicum (Liliaceae) inhabits deciduous mesic forests of Hokkaido, northern Japan. Myrmecochory of this species was investigated, especially the dispersal frequency, the effect of seed predators and the seed fall pattern. In the quadrat census using marked seeds of E. japonicum, the ant Myrmica kotokui frequently transported the seeds. However, the frequency of seed removal was low and most seeds were dispersed as little as 1 m or less. The spatial distribution of E. japonicum individuals was nearly random and most seedlings were established 5-20 cm away from the fertile plants, indicating that even this small scale of seed dispersal contributes to avoiding crowding of seedlings. Some arthropods, e.g. springtails, spiders and ticks, hindered seed dispersal by devouring elaiosomes and seeds. Although ground beetle species also damaged seeds and elaiosomes, a few of them exhibited seed removal behaviour. E. japonicum dropped their seeds not all at once but bit by bit, taking 3-6 days to drop all seeds. This seed-fall pattern was effective in raising the frequency of seed removal by ants and reducing seed predation by some arthropods.