Phylogenetic, population structure, and population demographic analyses reveal that Vicia sepium in Japan is native and not introduced.

Vicia sepium (bush vetch) is a perennial legume widely distributed throughout the Eurasian continent. However, its distribution in Japan is limited to Mt. Ibuki and small parts of central and southern Hokkaido. Therefore, each Japanese V. sepium lineage has been considered to have been introduced separately from Europe. Here, we examined whether the species was introduced or not on the basis of cpDNA sequences and genome-wide SNPs from Japanese and overseas samples. Both the cpDNA haplotype network and the nuclear DNA phylogenetic tree showed that Japanese V. sepium is monophyletic. Furthermore, although the nuclear DNA phylogenetic tree also showed that each lineage is clearly monophyletic, genetic admixture of the genetic cluster dominated in the Hokkaido lineage was also detected in the Mt. Ibuki lineage. Population divergence analysis showed that the two lineages diverged during the last glacial period. The Mt. Ibuki lineage showed a sudden population decline 300-400 years ago, indicating that some anthropogenic activity might be involved, while the Hokkaido lineage showed a gradual population decline from 5000 years ago. Consequently, these two lineages show low current genetic diversity compared with overseas lineages. These results show that the Japanese V. sepium is not introduced but is native.

Chemical composition and in vitro ruminal digestibility of Ludwigia grandiflora.

Ludwigia grandiflora is an amphibious plant that has been designated as a specific alien plant in Japan. Due to the risk of regeneration on land, plants are burned after eradication. Since L. grandiflora is fertile and rich in biomass, this study investigated the use of L. grandiflora as a feed for ruminants. We measured general components, detergent fiber components, and in vitro dry matter digestibility (IVDMD) using rumen microbes for three forms (terrestrial form, emergent form, and floating-leaved form) of L. grandiflora. In addition, concentrations of hazardous metallic elements were also measured from the viewpoint of feed safety. Crude protein, neutral detergent fiber, acid detergent fiber, and acid detergent lignin were contained 10.31-14.62, 27.83-42.28, 20.54-36.05, and 6.94-18.90 (%DM), respectively. IVDMD was 50.75%-68.24%. Toxic elements such as Pb, Cd, and Hg were not detected in all forms. These results suggest that L. grandiflora could be partially used as alternative roughage for ruminants.

Skewed male reproductive success and pollen transfer in a small fragmented population of the heterodichogamous tree Machilus thunbergii.

Heterodichogamy is defined as the presence of two flower morphs that exhibit the male and female functions at different times among individuals within a population. Heterodichogamy is regarded as an adaptation to promote outcrossing through enhanced inter-morph mating, together with a 1:1 morph ratio. However, in highly fragmented populations, the morph ratio may be more likely to be biased by stochastic events. In such a situation, individuals of a minority morph within a population are expected to have higher reproductive success than those of a majority morph, which may suffer from pollen shortages of the minority morph. In this paper, we evaluated mating patterns and male reproductive success in a highly fragmented population of Machilus thunbergii, a putative heterodichogamous evergreen laurel tree. Results of paternity analysis indicated that the selfing rate was not clearly different between the two morphs. In contrast, the proportion of intra-morph mating was higher in the majority-morph (MM) mother trees than in the minority-morph (MF) mother trees. Bayesian estimated male reproductive success indicated that male reproductive success was higher in minority-morph (MF) than in majority-morph (MM) mother trees. These findings indicate that (1) the majority morph mothers, suffering a shortage of the opposite morph pollen, could partly compensate for the reduced reproductive success by intra-morph mating rather than by selfing, and (2) negative-frequency dependent selection may be involved in the maintenance of the two morphs.

Range expansion and lineage admixture of the Japanese evergreen tree Machilus thunbergii in central Japan.

We investigated the range expansion histories of Machilus thunbergii populations in the Kinki region of central Japan on the basis of nuclear microsatellite data. In the Kinki region, M. thunbergii is typically found in the coastal area, with some fragmented populations inland, around Lake Biwa. Phylogenetic and Bayesian clustering analysis (STRUCTURE analysis) revealed that the inland populations have different genetic components between the west and east sides of Lake Biwa. The population located on the north side of the lake has an admixture of the two genetically differentiated lineages, contributing to an increase in the genetic diversity of the population. Populations around Lake Biwa had lost rare alleles and the F value obtained from STRUCTURE analysis was lower in the coastal populations than in the lake populations. These results suggest that populations around Lake Biwa experienced a bottleneck due to a founder effect during the initial migration to the lake and that glacial refugia of M. thunbergii in the Kinki region existed along the coast.

Development of microsatellites in Machilus thunbergii (Lauraceae), a warm-temperate coastal tree species in Japan.

PREMISE OF THE STUDY: Microsatellite markers were developed and characterized in a typically coastal, widespread, and dominant tree species of the evergreen broadleaf forests, Machilus thunbergii, for comparison of the genetic diversity and structure of inland populations surrounding the ancient Lake Biwa and coastal populations in Japan. METHODS AND RESULTS: Eighteen polymorphic microsatellites of this species were isolated using an improved technique for isolating codominant compound microsatellite markers. These isolated loci provided compound simple sequence repeat (SSR) markers with polymorphisms of three to 19 alleles per locus, with an average of 10.9. The expected and observed within-population heterozygosities ranged from 0.16 to 0.86 and from 0.13 to 0.72, respectively. CONCLUSIONS: These markers may be useful tools for further investigation of the population genetic structure and biogeographic history of M. thunbergii in the warm-temperate zone of East Asia.

Altitudinal and seasonal variations in the diet of Japanese macaques in Yakushima.

Altitudinal and seasonal variations in the diet of Japanese macaques in Yakushima, southwestern Japan, were studied for 2 years by means of fecal analysis. The altitudinal range of fecal samples collected was 30 m to 1,203 m above sea level, and it was divided into three zones: low-zone forest (0-399 m), middle-zone forest (400-799 m), and high-zone forest (800 m-1,230 m). There was a considerable altitudinal and seasonal variation in the macaques' diet. Seed/fruit and animal matter were eaten more in the lower zones, whereas more fiber and fungi were consumed in the higher zones. In all of the zones, they ate seed/fruits the most in autumn (September-November) and the least in spring (March-April). They ate fibrous food the most in spring and the least in autumn. Macaques relied on seed/fruits heavily in the lower zone for a longer period than in the higher zones. Macaques in the high-zone forest ate almost no seed/fruit foods from March to May. Altitudinal variations in availability of seed/fruit foods seem to have influenced the altitudinal variations in diet. Total basal area of seed/fruit-food trees, species richness of seed/fruit-foods, main seed/fruit-food types available, and annual fleshy-fruit production all decreased with increasing altitude. Both interannual variation and annual cyclicity of diet were found in all zones.

Interactions between fleshy fruits and frugivores in a tropical seasonal forest in Thailand.

Large frugivores are considered to be important seed dispersers for many tropical plant species. Their roles as seed dispersers are not well known in Southeast Asia, where degraded landscapes typically lack these animals. Interactions between 259 (65 families) vertebrate-dispersed fruits and frugivorous animals (including 7 species of bulbul, 1 species of pigeon, 4 species of hornbill, 2 species of squirrel, 3 species of civet, 2 species of gibbon, 1 species of macaque, 2 species of bear, 2 species of deer, and 1 species of elephant) were studied for 3 years in a tropical seasonal forest in Khao Yai National Park, Thailand. The purpose was to examine the dietary overlaps among the large frugivores and the characteristics of fruits they consumed. Most fruit species are eaten by various kinds of frugivores; no close relationship between a particular fruit and a frugivore was found. The number of frugivore groups that served a given plant species was negatively correlated with seed size. Additionally, the fruit/seed diameters consumed by bulbuls were significantly smaller than consumed by the other nine groups. These trends of fruit characteristics were consistent with those observed elsewhere in Southeast Asia: small fruits and large, soft fruits with many small seeds are consumed by a wide spectrum of frugivores while larger fruits with a single large seed are consumed by relatively few potential dispersers. Importantly, these large, single-seed fruits are not consumed by the small frugivores that thrive in small forest fragments and degraded areas in Southeast Asia. To insure the natural seed dispersal process in the forest, an evaluation of all frugivore groups in the forest is urgently needed in Southeast Asia.