Masashi Ohara

Effects of inflorescence size on visits from pollinators and seed set of Corydalis ambigua (Papaveraceae)

Female reproductive success (seed set) of a spring ephemeral plant, Corydalis ambigua Cham. et schlecht (Papaveraceae) was investigated in relation to inflorescence size and foraging behavior (frequency and duration of visitations) by pollinators (namely, overwintered queens of Bombus hypocrita sapporensis) by detailed daily observations of a natural population. Pollination experiments indicated that C. ambigua is self-incompatible and that seed set was significantly affected by the behavior of the pollinating queens. Plants with larger inflorescences were visited more often than those with fewer flowers. Fecundity also increased with increasing size of inflorescences. Visitation time (duration of foraging) rather than the frequency of visitations (number of visits) was critical for higher fecundity. Seed production was strongly enhanced by a few long visits (of more than 60 s), and seemed to be independent of large numbers of short visits (of less than 60 s). Hence, plants with larger inflorescences, which provide a conspicuous signal to pollinators and offer greater rewards in terms of nectar, received longer visits by B. hypocrita sapporensis queens and those plants exhibited higher fecundity.

Adaptive advantages of ant-dispersed seeds in the myrmecochorous plant Trillium tschonoskii (Liliaceae)

Trillium tschonoskii Maxim. is a myrmecochorous perennial herb which grows in single-species stands in the cool temperate broad-leaved deciduous woodland of Hokkaido, northern Japan. Its many-seeded fruit initially falls close to the parent, over 50% within 20 cm. Ants, principally Myrmica ruginodis and Aphaenogaster japonica, transport seeds to their nests, a mean distance of 64 cm. The nests are overdispersed and short-lived, and are no richer in nitrogen or phosphorus than surrounding soils. There are proportionally more older (3-leaved) than younger (1-leaved) juvenile Trillium plants at distances > 60 cm from the nearest parent plant than at 0-30 or 30-60 cm. This indicates that the relatively short-distance dispersal of T. tschonoskii seeds by ants is sufficient to reduce seedling mortality, by reducing competition between seedlings; and that this is the primary advantage of myrmecochory in this species. This contrasts with previous studies, involving ant species with longer-lived nests, where the main advantage to the plant is nutrient-enhanced seedling microsites.

Interference by Ground Beetles with the Dispersal by Ants of Seeds of Trillium Species (Liliaceae)

(1) Seed dispersal by ants (myrmecochory) was observed in populations of Trillium kamtschaticum and T. tschonoskii. In both populations, main agents of seed dispersal were Aphaenogaster smythiesijaponica and Myrmica ruginodiv, and the dispersal distance was 3 30 m (maximum) and 0 60 m on the average. (2) The seed removal frequency was only about 15%0, and the other 85 %X, of seeds were left near the mother plants, though all of them lost elaiosomes. (3) This loss was due to nocturnal ground beetles [Carabus arboreus, Aptomoterus japonicus (Carabinae), Pterostichus thunbergi, Synuchus melantho (Harpalinae), Silpha perforata venatoria (Silphidae) and Geotrupes laevistriatus (Scarabaeidae)] which were more active than ants at night and devoured the elaiosomes without transporting the seeds. (4) The seeds with the damaged elaiosomes usually did not attract ants. Thus, the ingestion of the elaiosomes by the ground beetles may cause a previously reported clumping of seedlings near fertile plants.

Genetic diversity and local population structure of fragmented populations of Trillium camschatcense (Trilliaceae)

Trillium camschatcense, a long-lived common woodland herb, has been experiencing intensive habitat fragmentation over the last century in eastern Hokkaido, Japan. We examined the genetic diversity and population genetic structure of 12 fragmented populations with different population sizes using allozyme electrophoresis. The percentage of polymorphic loci and mean number of alleles per locus were positively related to population size, probably due to the stochastic loss of rare alleles (frequency of q <0.1) in small populations. Populations with 350flowering plants or fewer had lost almost all of their rare alleles. While the heterozygosity and inbreeding coefficient were not related to population size, some small populations showed relatively high inbreeding coefficients. In spite of the low genetic differentiation among overall populations (FST=0.130), local population structuring was recognized between the two geographically discontinuous population groups. Within groups, sufficient historical gene flow was inferred, whereas a low dispersal ability of this species and geographical separation could produce apparent differentiation between groups. # 2002 Elsevier Science Ltd. All rights reserved.

Robber‐like pollinators: overwintered queen bumblebees foraging on Corydalis ambigua

ABSTRACT. 1 The behaviour of nectar‐collecting Bombus hypocrita sapporensis Cockerell queens was observed on a population of a spring ephemeral plant Corydalis ambigua Cham, et Schlecht. 2 Daily patterns of activity and behaviour changed with the progress of flowering. Activity peaked shortly before sunset early in the flowering season but approximately at noon towards the end of flowering. In the peak flowering period the queens tended to visit nearby plants and to change direction often, whereas early or late in the flowering period they flew further between visits and were less likely to change direction. 3 Each plant was visited 0 to 24 times (mean 9.4 ±SD 5.2) by the queens during the whole flowering season. 4 The queens collected nectar, rarely through the front of the flowers but mostly through the spurs perforated by themselves or predecessors. At the beginning of the flowering season the illegitimate foragers often visited the front of the flowers before moving to the spurs; later, most queens quickly learned to land directly on the spurs. 5 Even the 59.7% of plants that were visited only by illegitimate foragers set seeds. Close observation confirmed that the illegitimate foragers opened the inner petals enclosing anthers and stigma frequently when visiting the front of the flowers before robbing, or occasionally when walking about on the flowers or collecting nectar through the perforated spurs.

Seasonal changes in pollinator activity influence pollen dispersal and seed production of the alpine shrub Rhododendron aureum (Ericaceae)

In alpine ecosystems, microscale variation in snowmelt timing often causes different flowering phenology of the same plant species and seasonal changes in pollinator activity. We compared the variations in insect visitation, pollen dispersal, mating patterns, and sexual reproduction of Rhododendron aureum early and late in the flowering season using five microsatellites. Insects visiting the flowers were rare early in the flowering season (mid‐June), when major pollinators were bumblebee queens and flies. In contrast, frequent visitations by bumblebee workers were observed late in the season (late July). Two‐generation analysis of pollen pool structure demonstrated that quality of pollen‐mediated gene flow was more diverse late in the season in parallel with the high pollinator activity. The effective number of pollen donors per fruit (Nep) increased late in the season (Nep = 2.2–2.7 early, 3.4–4.4 late). However, both the outcrossing rate (tm) and seed‐set ratio per fruit were smaller late in the season (tm = 0.89 and 0.71, seed‐set ratio = 0.52 and 0.18, early and late in the season, respectively). In addition, biparental inbreeding occurred only late in the season. We conclude that R. aureum shows contrasting patterns of pollen movement and seed production between early and late season: in early season, seed production can be high but genetically less diverse and, during late season, be reduced, possibly due to higher inbreeding and inbreeding depression, but have greater genetic diversity. Thus, more pollinator activity does not always mean more pollen movement.

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.

Molecular Systematics of Trilliaceae I. Phylogenetic Analyses of Trillium Using matK Gene Sequences

matK was conducted using 41 Trillium taxa and two out-group taxa (Veratrum maackii and Helonias bullata). A total of 1608 base pairs were analyzed and compared., and then there were 61 variable (36 informative) sites among Trillium species. Fifteen insertion/deletion events (indels) of six or fifteen base pairs were also detected. Phylogenetic analyses of the sequence data revealed that the subgenus Phyllantherum (sessile-flowered species) forms a distinct monophyletic group, whereas the subgenus Trillium (pedicellate-flowered species) does not form a monophyletic group, and is composed of three distinct groups and three basally located species in the tree: (1) the Erectum group, (2) the Grandiflorum group, and (3) the Pusillum group and (4) the three species, including T. govanianum, T. undulatum, and T. rivale. T .rivale appears to be the most basally diverged and a very specialized taxon among the ingroup members. Our matK data indicated that the closest relative of the subgenus Phyllantherum is the Grandiflorum group. The results are concordant with the results of the RFLP analysis of cpDNA and also more or less with those of the cladistic analysis of morphological characters.

Variations in the breeding system and the population genetic structure of Trillium kamtschaticum (Liliaceae)

The breeding system of Trillium kamtschaticum (Liliaceae) was investigated in 23 populations located in Hokkaido, Japan. In addition, analyses of genetic structure were made based on allozyme variation. The populations in the eastern region of Hokkaido were found to exhibit high genetic variability. Emasculated individuals in these populations produced mature seeds, whereas bagged individuals did not. By contrast, northern and southern populations mostly exhibited very low genetic variability. Based on studies of chromosome variations in natural populations of this plant, it has been predicted that inbreeding is responsible for the low levels of genetic variation observed. However, in breeding experiments, both bagged and emasculated individuals produced mature seeds. Most of the self-compatible and genetically variable populations were at Hardy-Weinberg equilibrium. These results suggest that the plants in the eastern populations are self-incompatible and that seed production and high genetic diversity result from predominant outbreeding (insect pollination). By contrast, the northern and southern populations with low genetic diversity potentially exploit a mixed system of outbreeding and inbreeding.

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.