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How genetic diversity is maintained within a range of species is one of the crucial pieces of information in species conservation. Although the classical central-marginal hypothesis assumes genetic paucity in peripheral populations, this pattern is not always the case. The semi-shrub Chimaphila umbellata subsp. umbellate, population of which are located in the southernmost part of the subspecies distribution, is considered to be a threatened species in Japan. Thus, this study aimed to examine which populations should be preferentially conserved and if the central-marginal hypothesis can be applied to this case. Genetic diversity was examined in 15 populations of C. umbellata subsp. umbellata in Japan using 16 nuclear simple sequence repeat markers. Overall, the genetic diversity values within the populations did not correlate with the latitude of the population locality, although those of the southern marginal populations tended to be lower than those of populations in other regions. While the populations are genetically differentiated from each other, recent population size declines were only detected in a few cases. Bayesian inference of population structure revealed three genetically distinct groups. An approximate Bayesian computation revealed that these three genetic groups were derived from the ancient population in a recent times. The contribution of each population to total genetic diversity was estimated by removing a given population and recalculating the total genetic diversity. Only one population contributed to both gene diversity and allelic diversity, while several populations contributed to one or the other. Considering genetic diversity and structure, the conservation priority of Japanese populations of C. umbellata to preserve genetic diversity is discussed.

Understanding adaptation mechanisms is important in evolutionary biology. Parallel adaptation provides good opportunities to investigate adaptive evolution. To confirm parallel adaptation, it is effective to examine whether the phenotypic similarity has one or multiple origins and to use demographic modeling to consider the gene flow between ecotypes. Solidago yokusaiana is a rheophyte endemic to the Japanese Archipelago that diverged from Solidago virgaurea. This study examined the parallel origins of S. yokusaiana by distinguishing between multiple and single origins and subsequent gene flow. The haplotypes of noncoding chloroplast DNA and genotypes at 14 nuclear simple sequence repeat (nSSR) loci and single-nucleotide polymorphisms (SNPs) revealed by double-digest restriction-associated DNA sequencing (ddRADseq) were used for phylogeographic analysis; the SNPs were also used to model population demographics. Some chloroplast haplotypes were common to S. yokusaiana and its ancestor S. virgaurea. Also, the population genetic structures revealed by nSSR and SNPs did not correspond to the taxonomic species. The demographic modeling supported the multiple origins of S. yokusaiana in at least four districts and rejected a single origin with ongoing gene flow between the two species, implying that S. yokusaiana independently and repeatedly adapted to frequently flooding riversides.

Understanding the causes of the Sino‐Japanese disjunctions in plant taxa has been a central question in eastern Asian biogeography, with vicariance or long‐distance dispersal often invoked to explain such patterns. Diabelia Landrein (Caprifoliaceae; Linnaeoideae) comprises four shrubby species with a Sino‐Japanese disjunct distribution. The species diversification time within Diabelia, covering a long geological history of the formation process of the Sino‐Japanese flora, dated back to the middle Oligocene, therefore, Diabelia would be an ideal model to elucidate the biogeographic patterns of Sino‐Japanese disjunctions with climate fluctuation. In this study, we analyzed complete plastome sequence data for 28 individuals representing all four species of Diabelia. These 28 plastomes were found to be highly similar in overall size (156 243–157 578 bp), structure, gene order, and content. Our phylogenomic analysis of the plastomes supported a close relationship between Diabelia ionostachya (Nakai) Landrein & R.L. Barrett var. wenzhouensis (S.L. Zhou ex Landrein) Landrein from eastern China and Diabelia spathulata (Siebold & Zucc.) Landrein var. spathulata from Japan. Diabelia serrata (Siebold & Zucc.) Landrein was identified as sister to a population of Diabelia sanguinea (Makino) Landrein from Tochigi in central Japan and D. spathulata Landrein, from Toyama, central Japan. Most Diabelia lineages were estimated to have differentiated 8–28 Mya. Our results indicate that two independent vicariance events could explain the disjunction between Japan and Korea in the mid to late Miocene, and between Zhejiang and Japan in the early Miocene.

Edaphic specialization is one of the main drivers of plant diversification and has multifaceted effects on population dynamics. Carex angustisquama is a sedge plant growing only on heavily acidified soil in solfatara fields, where only extremophytes can survive. Because of the lack of closely related species in similar habitats and its disjunct distribution, the species offers ideal settings to investigate the effects of adaptation to solfatara fields and of historical biogeography on the genetic consequences of plant edaphic specialization to solfatara fields. Here, genome-wide single nucleotide polymorphisms were used to reveal the phylogenetic origin of C. angustisquama, and 16 expressed sequence tag-simple sequence repeat markers were employed to infer population demography of C angustisquama. Molecular phylogenetic analysis strongly indicated that C. angustisquama formed a monophyletic clade with Carex doenitzii, a species growing on nonacidified soil in the sympatric subalpine zone. The result of population genetic analysis showed that C. angustisquama has much lower genetic diversity than the sister species, and notably, all 16 loci were completely homozygous in most individuals of C. angustisquama. Approximate Bayesian computation analysis supported the model that assumed hierarchical declines of population size through its evolutionary sequence. We propose that the edaphic specialist in solfatara fields has newly attained the adaptation to solfatara fields in the process of speciation. Furthermore, we found evidence of a drastic reduction in genetic diversity in C. angustisquama, suggesting that the repeated founder effects associated with edaphic specialization and subsequent population demography lead to the loss of genetic diversity of this extremophyte in solfatara fields.

Of the two major speciation modes of endemic plants on oceanic islands, cladogenesis and anagenesis, the latter has been recently emphasized as an effective mechanism for increasing plant diversity in isolated, ecologically homogeneous insular settings. As the only flowering cherry occurring on Ulleung Island in the East Sea (concurrently known as Sea of Japan), Prunus takesimensis Nakai has been presumed to be derived through anagenetic speciation on the island. Based on morphological similarities, P. sargentii Rehder distributed in adjacent continental areas and islands has been suggested as a purported continental progenitor. However, the overall genetic complexity and resultant non-monophyly of closely related flowering cherries have hindered the determination of their phylogenetic relationships as well as the establishment of concrete continental progenitors and insular derivative relationships. Based on extensive sampling of wild flowering cherries, including P. takesimensis and P. sargentii from Ulleung Island and its adjacent areas, the current study revealed the origin and evolution of P. takesimensis using multiple molecular markers. The results of phylogenetic reconstruction and population genetic structure analyses based on single nucleotide polymorphisms detected by multiplexed inter-simple sequence repeat genotyping by sequencing (MIG-seq) and complementary cpDNA haplotypes provided evidence for (1) the monophyly of P. takesimensis; (2) clear genetic differentiation between P. takesimensis (insular derivative) and P. sargentii (continental progenitor); (3) uncertain geographic origin of P. takesimensis, but highly likely via single colonization from the source population of P. sargentii in the Korean Peninsula; (4) no significant reduction in genetic diversity in anagenetically derived insular species, i.e., P. takesimensis, compared to its continental progenitor P. sargentii; (5) no strong population genetic structuring or geographical patterns in the insular derivative species; and (6) MIG-seq method as an effective tool to elucidate the complex evolutionary history of plant groups.

[EN] Reviews that summarize the genetic diversity of plant species in relation to their life history and ecological traits show that forest trees have more genetic diversity at population and species levels than annuals or herbaceous perennials. In addition, among-population genetic differentiation is significantly lower in trees than in most herbaceous perennials and annuals. Possible reasons for these differences between trees and herbaceous perennials and annuals have not been discussed critically. Several traits, such as high rates of outcrossing, long-distance pollen and seed dispersal, large effective population sizes (N), arborescent stature, low population density, longevity, overlapping generations, and occurrence in late successional communities, may make trees less sensitive to genetic bottlenecks and more resistant to habitat fragmentation or climate change. We recommend that guidelines for genetic conservation strategies be designed differently for tree species versus other types of plant species. Because most tree species fit an LH scenario (low [L] genetic differentiation and high [H] genetic diversity), tree seeds could be sourced from a few populations distributed across the species’ range. For the in situ conservation of trees, translocation is a viable option to increase N. In contrast, rare herbaceous understory species are frequently HL (high differentiation and low diversity) species. Under the HL scenario, seeds should be taken from many populations with high genetic diversity. In situ conservation efforts for herbaceous plants should focus on protecting habitats because the typically small populations of these species are vulnerable to the loss of genetic diversity. The robust allozyme genetic diversity databases could be used to develop conservation strategies for species lacking genetic information. As a case study of reforestation with several tree species in denuded areas on the Korean Peninsula, we recommend the selection of local genotypes as suitable sources to prevent adverse effects and to insure the successful restoration in the long term., [ES] Las revisiones que resumen la diversidad genética de las plantas en relación con sus características ecológicas y biológicas muestran que los árboles forestales tienen más diversidad genética a nivel de población y de especie que las plantas anuales o las perennes herbáceas. Sumado a esto, la diferenciación genética entre poblaciones es significativamente más baja en los árboles que en la mayoría de las perennes herbáceas y las anuales. Hasta la fecha no se han discutido críticamente las posibles explicaciones de estas diferencias entre los árboles y las perennes herbáceas y las plantas anuales. Varias características, como las tasas altas de alogamia, la dispersión a gran distancia de polen y semillas, el gran tamaño de la población efectiva (Ne), la estatura arbórea, la baja densidad poblacional, la longevidad, el solapamiento de generaciones y la presencia dentro de comunidades sucesionales tardías, pueden generar en los árboles una menor sensibilidad a los cuellos de botella genéticos y una mayor resistencia a la fragmentación del hábitat o al cambio climático. Recomendamos que las directrices para las estrategias de conservación genética estén diseñadas de manera diferente para las especies arbóreas que para otro tipo de plantas. Ya que la mayoría de las especies arbóreas encajan dentro de un escenario LH (baja [L] diferenciación genética y alta [H] diversidad genética), las semillas de los árboles podrían tomarse de unas cuantas poblaciones dispersas a lo largo del área de distribución de la especie. Por lo anterior, para la conservación in situ de los árboles, la translocación es una opción viable para incrementar la Ne. Al contrario de esta situación, las especies herbáceas raras del sotobosque con frecuencia son especies HL (alta diferenciación y baja diversidad). En el escenario HL, las semillas deberían ser recolectadas de muchas poblaciones con diversidad genética alta y los esfuerzos de conservación in situ para las plantas herbáceas deberían enfocarse en la protección del hábitat ya que las poblaciones típicamente pequeñas de estas especies son vulnerables a la pérdida de la diversidad genética. Las robustas bases de datos de diversidad genética aloenzimática podrían usarse para desarrollar estrategias de conservación para las especies que carecen de información genética. Como caso de estudio de reforestación con varias especies arbóreas en áreas deforestadas de la Península de Corea, recomendamos la selección de genotipos locales como fuente adecuada para prevenir los efectos adversos y para asegurar la restauración exitosa a largo plazo., This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT of the Republic of Korea (NRF-2013R1A1A2063524 to M.Y.C. and NRF-2017R1A2B4012215 to M.G.C.), Introduction Higher Genetic Diversity of Trees than Herbaceous Perennials or Annuals Strategies for Conserving Genetic Diversity in Trees and Herbaceous Species Case Study of Restoration of a Denuded Area Conclusions Acknowledgments

BACKGROUND AND AIMS: The processes and mechanisms underlying lineage diversification are major topics in evolutionary biology. Eurasian goldenrod species of the Solidago virgaurea complex show remarkable morphological and ecological diversity in the Japanese Archipelago, with ecotypic taxa well adapted to specific environments (climate, edaphic conditions and disturbance regimes). The species complex is a suitable model to investigate the evolutionary processes of actively speciating plant groups, due to its ability to evolve in relation to environmental adaptation and its historical population dynamics. METHODS: Two chloroplast markers, 18 nuclear microsatellite markers and ddRAD-sequencing were used to infer population genetic demography of S. virgaurea complex with its related species/genera. KEY RESULTS: Our analysis showed that populations in Japan form an evolutionary unit, which was genetically diverged from adjacent continental populations. The phylogenetic structure within the archipelago strongly corresponds to the geography, but interestingly there is no concordance between genetic structure and ecotypic boundaries; neighbouring populations of distinct ecotypes share a genetic background. CONCLUSIONS: We propose that the traits specific to the ecotypic entities are maintained by natural selection or are very recently generated and have little effect on the genomes, making genome-wide genetic markers unsuitable for detecting ecotypic differentiation. Furthermore, some sporadically distributed taxa (found as rheophytes and alpine plants) were repeatedly generated from a more widespread taxon in geographically distant areas by means of selection. Overall, this study showed that the goldenrod complex has a high ability to evolve, enabling rapid ecological diversification over a recent timeframe.

South Asian country Nepal characterizes a complex mountain range in this world. The country’s population density is increasing along with rapid growth of population especially over mountainous cities, southern hills and the Terai. On the other hand, a number of fatal natural calamities, such as flash flooding and landslides raised by clutter intensive rainfall, have been increasing since the last decade. To deal with such water hydro meteorological disasters, accurate information on spatial and temporal variation of rainfall distribution is very important. In Nepal, the amount of rainfall has been obtained from limited rain gauge networks, which may leads to many errors in making a Quantitative Precipitation Estimation (QPE). Weather radar observations have recently been highlighted as an alternative option for estimating the spatial and temporal distribution of precipitation across specified time intervals. However, estimating rainfall from radar observation has its own challenges, especially over a mountainous country like Nepal. Another mountainous country Japan is well known for using weather radar observation to make QPE product. Different types of weather radar have been used to record, monitor and forecast precipitation in Japan for both operational and research purposes. A high level research work has also been done on this field. The high spatial and temporal (250-m and 1-min) QPE product obtained from the radar observation is available for the public. It shows good harmony with ground data in the flat and mountain areas of Japan. Though Nepal and Japan are located in different regions, both countries represent complex mountain regions and have been facing natural disaster caused by extreme rainfall. In Nepal, weather radar observation for estimating precipitation amounts has not started on an operational basis till date. Hence, sharing knowledge and skills from Japan’s research on weather radar observation would play a key role to achieve the radar based QPE product in Nepal. Therefore, we discuss about the challenge in obtaining QPE product, considering an example of the progress of weather radar system in Japan. It is believed that any discussion on it will be a reference for weather radar deployment and its QPE product in Nepal in coming days. HYDRO Nepal Journal Journal of Water Energy and Environment Issue: 21, July, 2017 Page: 50-59 Upload Date: July 18, 2017

The Sino-Japanese Floristic Region (SJFR) is a key area for plant phylogeographical research, due to its very high species diversity and disjunct distributions of a large number of species and genera. At present, the root cause and temporal origin of the discontinuous distribution of many plants in the Sino-Japanese flora are still unclear. Diabelia (Caprifoliaceae; Linnaeoideae) is a genus endemic to Asia, mostly in Japan, but two recent discoveries in China raised questions over the role of the East China Sea (ECS) in these species' disjunctions. Chloroplast DNA sequence data were generated from 402 population samples for two regions (rpl32-trnL, and trnH-psbA) and 11 nuclear microsatellite loci were screened for 549 individuals. Haplotype, population-level structure, combined analyses of ecological niche modeling, and reconstruction of ancestral state in phylogenies were also performed. During the Last Glacial Maximum (LGM) period after the Tertiary, Diabelia was potentially widely distributed in southeastern China, the continental shelf of the East China Sea and Japan (excluding Hokkaido). After LGM, all populations in China have disappeared except those in Zhejiang which may represent a Glacial refuge. Populations of Diabelia in Japan have not experienced significant bottleneck effects, and populations have maintained a relatively stable state. The observed discontinuous distribution of Diabelia species between China and Japan are interpreted as the result of relatively ancient divergence. The phylogenetic tree of chloroplast fragments shows the characteristics of multi-origin evolution (except for D. sanguinea). STRUCTURE analysis of nuclear Simple Sequence Repeat (nSSR) showed that the plants of the Diabelia were divided into five gene pools: D. serrata, D. spathulata, D. sanguinea, D. ionostachya (D. spathulata var. spathulata-Korea), and populations of D. ionostachya var. ionostachya in Yamagata prefecture, northern Japan. Molecular evidence provides new insights of Diabelia into biogeography, a potential glacial refuge, and population-level genetic structure within species. In the process of species differentiation, ECS acts as a corridor for two-way migration of animals and plants between China and Japan during glacial maxima, providing the possibility of secondary contact for discontinuously distributed species between China and Japan, or as a filter (creating isolation) during glacial minima. The influence of the ECS in speciation and biogeography of Diabelia in the Tertiary remains unresolved in this study. Understanding origins, evolutionary histories, and speciation will provide a framework for the conservation and cultivation of Diabelia.

To determine the origin and genetic consequences of anagenesis in Rubus takesimensis on Ulleung Island, Korea, we compared the genetic diversity and population structure of R. takesimensis with those of its continental progenitor R. crataegifolius. We broadly sampled a total of 315 accessions in 35 populations and sequenced five noncoding regions of chloroplast DNA. Rubus takesimensis emerged as nonmonophyletic and several geographically diverse continental populations were likely responsible for the origin of R. takesimensis; the majority of R. takesimensis accessions were sisters to the clade containing accessions of R. crataegifolius, primarily from the Korean peninsula, while rare accessions from three populations shared common ancestors with the ones from the southern part of the Korean peninsula, Jeju Island, and Japan. A few accessions from the Chusan population originated independently from the Korean peninsula. Of 129 haplotypes, 81 and 48 were found exclusively in R. crataegifolius and R. takesimensis, respectively. We found unusually high genetic diversity in two regions on Ulleung Island and no geographic population structure. For R. crataegifolius, two major haplotype groups were found; one for the northern mainland Korean peninsula, and the other for the southern Korean peninsula and the Japanese archipelago. Compared with populations of R. crataegifolius sampled from Japan, much higher haplotype diversity was found in populations from the Korean peninsula. The patterns of genetic consequences in R. takesimensis need to be verified for other endemic species based on chloroplast DNA and independent nuclear markers to synthesize emerging patterns of anagenetic speciation on Ulleung Island.

Cremastra appendiculata var. variabilis is a self-compatible, insect-pollinated, terrestrial orchid that is a typical member of the warm-temperate vegetation in the Korean Peninsula. Here we examine levels and partitioning of allozyme diversity (22 loci) in 12 populations of this orchid to gain insight into its genetic structure and post-glacial colonization history in Korea. It harboured considerably higher levels of genetic variation within populations (%P = 48.1, A = 1.70 and He = 0.217) and lower degree of differentiation among populations (FST = 0.068) than those typical of allozyme-based studies in other terrestrial orchid species. These patterns suggest that extant populations were derived from multiple source populations (i.e. from multiple glacial refugia), although further studies are needed to confirm this scenario. In addition to population history, traits such as high potential of seed dispersal, a mixed mating system and its occurrence in large and continuous populations would have contributed to the current levels and distribution of genetic diversity in Korean populations of C. appendiculata var. variabilis. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 721–732.

Compared to tree species, there still remains a very limited number of phylogeographical studies on temperate forest perennials. In the present study, we used nuclear microsatellite markers to detect extant patterns of population genetic structure of the perennial plant Veratrum album ssp. oxysepalum in the understory of temperate forests, aiming to provide insights into the post-glacial history of the species and the reasons for the disjunct distribution of chloroplast DNA (cpDNA) haplotypes found in a previous study. We examined eight polymorphic nuclear microsatellite loci to examine genetic variation within and among 32 populations of V. album ssp. oxysepalum and seven populations of Veratrum stamineum. The population of V. album ssp. oxysepalum was split into two groups: those in the northern part of the Japanese Archipelago and all others. In addition, each population group was split into two subgroups. Only one population of V. stamineum included a substantial number of individuals categorized as V. album ssp. oxysepalum. The population genetic structure revealed by nuclear microsatellites suggests that V. album ssp. oxysepalum migrated from Eurasia into the Japanese Archipelago at least twice before the last glacial era through two different land bridges: the Korean Peninsula and Sakhalin. In addition, there appeared to be at least two refugia of V. album ssp. oxysepalum in each of the areas, dominated by the two different lineages from Eurasia during the last glacial era. The disjunct geographical pattern found in cpDNA variation in a previous study was blurred in the genetic structure as revealed by microsatellite markers, probably as a result of extensive gene flow via pollen. Currently, one-directional introgressive hybridization occurs from V. album ssp. oxysepalum to V. stamineum at only one locality; there is no evidence of the opposite pattern. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 108, 278–293.

It is thought that the warm-temperate vegetation of Korea, which at present is limited to southern coastal areas, was founded by individuals from glacial refugia putatively located in southern Japan and/or southern China. Two scenarios of postglacial recolonization can be hypothesized: 1) extant Korean populations are derived from multiple source populations or, 2) they originated from a single source. To test which of these scenarios is more likely, we surveyed patterns of genetic diversity in 3 congeneric terrestrial orchids that are typical of Korean warm-temperate vegetation, Calanthe discolor, C. sieboldii, and C. reflexa. In total, we studied 14 populations with 17 allozyme loci. To complement the study, we also tested the 2 scenarios with previous population-genetics data reported for other warm-temperate plant species native to Korea. Levels of genetic variation in the 3 Calanthe species were substantially higher than those typical of allozyme-based studies in other terrestrial orchid species, which clearly suggest a pattern of postglacial recolonization fitting the first scenario. However, pre vious studies with native Korean species showed equivocal patterns and suggest conflicting scenarios. The Calanthe species studied here and the previous genetic surveys conducted on other species indicate that the establishment of contemporary warm-temperate vegetation in Korea has been diverse since deglaciation. From a conservation perspective, C. reflexa should be regarded as endangered in Korea, needing special attention to preserve its current genetic diversity and to prevent further decreases in population sizes.

We hypothesized that the main Korean mountain ranges provided many refugia for boreal plant species, where they likely found relatively stable habitats and maintained large population sizes. Under this scenario, high levels of genetic variation and low degree of differentiation among populations within these species were anticipated. To test this hypothesis, we examined levels of allozyme diversity (17 loci) in 12 populations of the common terrestrial montane orchid Oreorchis patens from the main ranges in Korea and 4 populations of its rare congener O. coreana, which is restricted to the Korean island of Jeju. As expected, O. patens harbored high levels of genetic variation within populations (%P = 62.8, A = 1.96, H (o) = 0.211, and H (e) = 0.237). Allele frequency differences among populations were low (F (ST) = 0.075), and the species also displayed a significant correlation between pairwise genetic differentiation and geographical distance. All these results suggest that extant populations were founded by multiple genetically diverse individuals and that most of this initial diversity would have been maintained in the stable mountainous conditions during Quaternary climatic oscillations. In contrast, we were unable to detect any genetic diversity in O. coreana, suggesting that contemporary populations likely originated from a single ancestral source population that had lost all genetic variability. From a long-term conservation genetics perspective, extreme rarity and small population sizes, coupled with its apparent genetic uniformity, place O. coreana at a high risk of extinction. Thus, both in situ and ex situ conservation efforts should be of particular importance for this species.

Aim To provide insights into genetic differentiation between insular endemic Weigela coraeensis var. fragrans and its progenitor variety W. coraeensis var. coraeensis, the population genetic structure of both varieties was examined, and factors promoting genetic differentiation between the two taxa were explored. Location The natural range of W. coraeensis (sensu lato) throughout mainland Japan (Honshu) and the Izu Islands. Methods The analysis included 349 and 504 individuals across the mainland (Honshu) and the Izu Islands, respectively, using 10 allozyme and 10 microsatellite loci. The population genetic structure of W. coraeensis was assessed by analysing genetic diversity indices for each population, genetic differentiation among populations, model-based Bayesian clustering or distance-based clustering, and bottleneck tests. Results The level of genetic diversity in each of the populations on the Izu Islands was negatively correlated with geographical distance between each island and the mainland. The populations on the mainland and on the Izu Islands were genetically differentiated to a certain extent; however, the microsatellite analyses suggested that gene flow also occurred between the mainland and the islands, and among individual islands. These microsatellite analyses also suggested recent bottlenecks in several populations in both areas. Main conclusions The decrease in genetic diversity throughout the Izu Islands, which correlated with distance to the mainland, Honshu, may be the result of a repeated founder effect occurring at a series of inter-island colonizations from north to south. The stepping stone-like configuration of the islands may have played a role in the dispersal of the species. Geographical isolation by sea would effectively result in genetic differentiation of W. coraeensis between mainland Honshu and the Izu Islands, although some gene flow may still occur between Honshu and the northern Izu Islands. The differentiation process of the endemic plants on the Izu Islands is anagenetic but not completed, and the study of these plants will provide insightful knowledge concerning the evolution of insular endemics.

Premise of the study Expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed for Carex angustisquama (Cyperaceae) to investigate the evolutionary history of this plant that is endemic to solfatara fields in northern Japan. Methods and results Using RNA-Seq data generated by the Illumina HiSeq 2000, 20 EST-SSR markers were developed. Polymorphisms were assessed in C. angustisquama and the closely related species C. doenitzii and C. podogyna. In C. angustisquama, many loci were monomorphic within populations; the average number of alleles ranged from one to five, and levels of expected heterozygosity ranged from 0.000 to 0.580, while all markers were polymorphic in a population of C. doenitzii. This indicates that low genetic polymorphism of C. angustisquama is likely due to the species' population dynamics, rather than to null alleles at the developed markers. Conclusions These markers will be used to assess genetic diversity and structure and to investigate evolutionary history in future studies of C. angustisquama and related species.

Allozyme diversity was examined in 30 populations of the maritime perennial plant Hedyotis strigulosa var. parviflora, which is distributed from subtropical islands to the central mainland of Japan. Genetic diversity within populations tended to be larger in southern island populations than in northern mainland populations. In the southern part of the distribution, the population size is generally large and populations are distributed more continuously than in the northern area, resulting in the larger effective size of southern populations as a whole. These factors play a major role in maintaining greater genetic diversity in the southern populations. By contrast, genetic diversity in the northern populations is very low, probably resulting from bottlenecks of population establishments during recolonization from refugial area to the northern areas. Geographically close populations were located near each other in the multidimensional scaling and the phenogram based on genetic distances, suggesting that gene flow among remote populations is rather limited. The pattern of genetic diversity in H. strigulosa var. parviflora is likely caused by the distribution expansion of the species; in the last glacier era, the species was restricted to the southern area; its advance to the northern area is relatively recent. Another variety endemic only to the Daito Islands, H. strigulosa var. luxurians, has lower genetic diversity than H. strigulosa var. parviflora and has genetically diverged from H. strigulosa var. parviflora. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 93, 679–688.

We examined the genetic diversity and structure in populations of the endangered grassland herb Vincetoxicum atratum using 11 polymorphic microsatellite loci. Although the populations were small and disconnected, our molecular data indicated that the species maintains relatively high levels of genetic diversity and connectivity among populations. Population clustering analyses detected 2 to 3 clusters and most of the populations of V. atratum comprised admixture of these genetic clusters. These admixtures likely formed during the process of colonizing habitats that had been disturbed by human activities. However, STRUCTURE clustering detected low-admixtures in populations occurring in rocky maritime sites, which may not be suitable for agriculture/rangeland activities. High genetic diversity and population connectivity suggested that loss of the remaining populations by grassland reduction might be an immediate threat for this species. Small grasslands populations managed by local farmers need appropriate conservation practices. Although our results showed genetic diversity and gene flow among populations of V. atratum were high, it is possible that this resulted from the historical continuous distribution of the species. To examine this hypothesis, further periodical monitoring of the genetic diversity and the genetic differentiation for the species is needed for a conservation action of the species.

Premise of the study: Microsatellite markers were developed in Veratrum maackii (Melanthiaceae) to assess the pattern of population genetic structures across the species' distribution. Methods and Results: Ten polymorphic loci were isolated from V. maackii using 454 shotgun pyrosequencing and the biotin capture method. The number of alleles per locus ranged from three to 19. The observed heterozygosity and expected heterozygosity ranged from 0.143 to 0.929 and 0.269 to 0.904, respectively. Conclusions: These newly developed microsatellite markers are useful for assessing the genetic diversity, population structure, and demographic history of V. maackii across its distribution range.

Reproductive character and genetic diversity were investigated for an autogamous species of Tylophora matsumurae endemic to the Ryukyu Islands and its progenitor species T. tanakae. Approximately one-fourth of bagged flowers set fruits in T. matsumurae, although no fruits were obtained from bagged flowers in T. tanakae. In situ pollen tube germination was observed with high frequency in all five populations examined in T. matsumurae. Furthermore, in T. matsumurae, anther sacs were not dehisced even at anthesis. These results suggest the highly autogamous nature of T. matsumurae. No allozymic variation was detected in all seven populations examined in T. matsumurae. The phenogram constructed using the neighbor-joining method based on Nei's unbiased genetic distance indicated that T. matsumurae clustered with the Okinoerabu Island population of T. tanakae. The estimated outcrossing rate of four populations of T. tanakae varied from 0.18 to 0.59. It is probable that T. matsumurae had been derived from the predominantly self-pollinating population of T. tanakae, and rapidly enlarged its distribution area.

Results of studies using molecular markers for determining demographic and genetical population parameters especially in plants or sessile animals under field conditions are strongly dependent on the sampling strategy adopted. There are two critical decisions to make when determining this strategy: (i) what is the unit to be sampled?, (ii) how should units to be sampled in the field be selected? For the first decision, there are two conceptually different approaches: sampling ramets of clonal plants as units (to get information about within-genet parameters, such as genet sizes or numbers) and sampling genets of clonal or non-clonal plants as units (to get information of the genetic structure of the population). For the second decision, it is critically important to make the goal of the study explicit. We argue that in this case fully random sampling is needed only when an estimate of the true value of the population parameter is needed; if a comparison between populations is the goal, however, other sampling schemes may be adopted. The efficiency of different types of sampling strategies to recover relative values in a spatially extended population is studied by means of a spatially explicit simulation model. The results show that a regular pattern of sampling is best for obtaining information on genet sizes or inbreeding coefficients; in contrast, random or hierarchical sampling strategies are better for obtaining information on parameters that are based on comparison of pairs of individuals, such as distribution of genet sizes or autocorrelation in genetic structure. A set of recommendations is provided for designing a good sampling strategy.

Genetic diversity and genetic differentiation within and among island populations was examined by allozyme electrophoresis in Suzukia luchuensis (Labiatae), which is endemic to four of the Ryukyu Islands, southern Japan, and one island near Taiwan. Intrapopulation allozyme diversity was very low in all the four Ryukyu Islands, probably due to the effects of random drift in small populations. In contrast, genetic diversity at the species level was high, possibly because of an ancient origin of populations and/or multiple colonization of the species on different islands. Genetic differentiation among the overall populations was high (G(ST)=0.863), while gene flow (Nm) as estimated from allozyme frequency data was 0.041, suggesting that its occurrence among populations is highly restricted. Hierarchical analysis of genetic differentiation indicated that a high proportion of the total allelic variance is attributed to variation among islands, corresponding to the fact that several alleles were fixed on only one island. However, intraisland genetic differentiation was small on all islands except Yonaguni Island, where S. luchuensis is relatively widely distributed. Most diversity was thus due to differences among islands.

Menziesia goyozanensis, a Japanese kindof fool's huckleberry, is a shrub speciesendemic only to Mt. Goyo in northern Japan.This species is listed as `criticallyendangered' in the Red Data Book of wild plantsin Japan, and only a single population has yetbeen discovered. The allozyme diversity andRAPD variation of Menziesia pentandrawere eight- and threefold greater than those ofM. goyozanensis, respectively, althoughwe examined only five populations representinga part of the distribution of M.pentandra. Because the environment surroundingthe population of M. goyoznaensis seemsto be stable at present, it is not necessary toplan an urgent ecological conservation practicefor the species. However, considering that thehabitat of M. goyozanensis is very smalland large vegetative disturbances occasionallyoccur in cool temperate forests, in which thespecies dwells, ex situ conservation isessential.

Veratrum stamineum var. micranthum is a perennial herb listed in the Red Data Book of Japan that is geographically restricted to central Japan. We developed 10 polymorphic microsatellite markers to examine genetic variability in this species. The observed and expected heterozygosities ranged from 0.333–1.000 and 0.370–0.898, respectively. Two of the 10 loci significantly deviated from Hardy–Weinberg equilibrium, suggesting biparental inbreeding due to population size contraction or existence of null alleles. These markers will be useful in population genetic studies and for the preservation of V. stamineum var. micranthum.

RAPD variation was examined in nine populations of Campanula microdonta Koidz., endemic to the Izu Islands, Japan. Ninety-eight bands were obtained for all populations, 94% of which were polymorphic at least within a population. Shannon's H values were calculated; these have frequently been used in RAPD studies to estimate genetic diversity. The values within populations did not correlate with the allozyme gene diversity estimated by a previous study or with distance from the Japanese mainland. The possible reasons for this discrepancy are different selection regimes between the two markers, higher RAPD mutation rates, and each marker's different coverage of genomes. Cluster analysis of genetic similarities suggested that colonization of each island probably occurred once, except for Miyake Island, where immigration has occurred at least twice.

Populations of each of the four species of Tricyrtis sect. Flavae were sampled using enzyme electrophoresis to examine the effect of geographic range and dichogamy on the genetic diversity of the species. The most widespread species, T. nana, had the lowest level of genetic diversity at both the population and the species level. The depauperate genetic diversity at the population level of T. nana appears to result from the high self-fertilization of the species. The low genetic diversity at the species level of T. nana probably resulted from the bottleneck effect during the speciation process in which this species diverged from the progenitor species, T. flava . Genetic differentiation among populations was high in both adichogamous T. nana and protandrous T. flava . High self-fertilization in T. nana and the colonizing nature of T. flava are likely the main factors causing the differentiated population genetic structure. In contrast to a previous study on chloroplast DNA (cpDNA) variation in Tricyrtis sect. Flavae, T. nana was most closely related to T. flava , which corresponds to the morphological resemblance of both species.

Ligustrum ovalifolium is a semi-evergreen tree distributed in the western part of Japan and southern Korea. This species contains an insular endemic variety, L. ovalifolium var. pacificum; this variety occurs only on the Izu Islands located south of the Japanese mainland Honshu. We isolated 10 polymorphic microsatellite loci from the species, and characterized them for 21 individuals from a population of L. ovalifolium. The primers developed in this study yielded an average 12.2 alleles per locus and an average expected heterozygosity of 0.78. These markers will be powerful tools for studying the genetic differentiation within the species.

Eight microsatellite markers were developed for the endangered grassland perennial herb Vincetoxicum atratum. The number of alleles ranged from 4 to 14, and the expected heterozygosities were from 0.575 to 0.933 in a population of V. atratum. Five of the eight loci did not significantly deviated from the Hardy–Weinberg equilibrium. All eight loci were tested for cross-species amplification in five other species of Vincetoxicum in Japan. These microsatellite loci will be useful for conservation genetics of V. atratum and other species of Vincetoxicum.

Hostas are very popular ornamental plants in gardens in the United States, Europe, and East Asia. We have developed nine microsatellite loci from an enrichment library of genomic DNA in Hosta albomarginata. We characterized these nine microsatellite loci for 20 individuals from a population of H. albomarginata. The primers developed in this study yielded an average of 12.4 alleles per locus (range five to 20) and an average expected heterozygosity of 0.86 (range 0.65 to 0.95). These markers will be powerful tools for breeding programmes of hosta cultivars, studies of population genetics, and the conservation of wild hosta species.

Twelve polymorphic microsatellite loci were isolated and characterized from the threatened toad lily Tricyrtis flava. The number of alleles ranged from three to 17, and the observed and expected heterozygosities ranged from 0.35 to 0.96 and from 0.50 to 0.93, respectively, in a population of T. flava. We also tested cross-species amplifications of the markers in three other species of Trycyrtis sect. Flavae. Most of 12 markers developed for T. flava were successfully amplified in the species of sect. Flavae and will be useful for population genetic studies of these species.

To identify the factors responsible for degrading the habitat of the endangered plant species Aster kantoensis, as well as the vulnerable life stage where this occurs, we carried out sowing experiments. Two natural habitats were simulated, being situated along the floodplains of the Tama River in central Japan. Seeds collected from a natural habitat were sown in two apparently suitable locations (Tomoda and Ishida sites). Germination, survival, growth, and seed production were subsequently monitored from 1993 through to 1997. The Tomoda site was a gravel bar in floodplains formed by flooding in 1991, while the Ishida site (two plots) was one gravel bar where several plants were growing sparsely and another where a population had become extinct in 1992. Seed cohorts completed their life cycle within 3 years at the Ishida site and within 5 years at the Tomoda site. Monitored parameters at Ishida were substantially lower than those at Tomoda. In addition, estimates of population growth indicated an increase at Tomoda and a rapid decrease at Ishida. However, degradation of habitats seemed to occur at Tomoda over the monitored periods. In view of our results, we conclude that natural germination of about 0.13% is needed for increasing population size. The major factors for decreasing population size are believed to be the lack of safe sites for germination and seedling establishment in old habitats (> 10 years). Conservation measures are suggested based on these findings.

Genetic variability was examined at 16 putative allozymic loci in the narrow endemic fern Polystichum otomasui. Although this species is distributed in only a few valleys within an approximately 10 km×6 km area on the Kyusyu Island of Japan, there are a relatively large number of individuals in this area. The percentage of polymorphic loci (P), the number of alleles per locus (A), the observed heterozygosity (HO) and the expected heterozygosity (HE) were 61.9, 1.93, 0.166 and 0.177, respectively, at the population level, and P, A and HE were 81.3, 2.69 and 0.199, respectively, at the species level, indicating that P. otomasui has extremely high allozymic variation for a fern species. The GST value among the 10 populations examined within the range was 0.108, suggesting that the interpopulation gene flow is high enough to impede genetic structuring. The mating system estimates, Wright's fixation index and the intragametophytic selfing rate, indicate that the species is outcrossing. High genetic variability, in spite of narrowness of the distribution, may have resulted from a recent rapid decline in the population and/or the large effective population size resulting from the extensive gene flow among the populations.

Genetic variation in an endangered tree species, Chionanthus retusus, was examined using enzyme electrophoresis. This species occurs in East Asia from Japan to Taiwan and southern China. In Japan, however, it occurs only in two restricted areas: at the northernmost tip of Tsushima Island between Japan and Korea, and in the eastern Gifu Prefecture (Tono region) of central Honshu. In the Tono region, the habitat of C. retusus is becoming fragmented under the influence of human activity and each stand contains a few individuals of C. retusus, while the population on Tsushima consists of approximately 1000 individuals. From the fossil evidence, C. retusus has been distributed continuously including the both two regions, and the present distribution is apparently relic in Japan. In contrast to the Tono population, the population of Tsushima seems to be stable. Comparing the genetic diversity and differentiation of the populations between these two regions should shed light on how a relic woody long-lived plant keeps and/or looses its genetic diversity, and how a population differentiates from other disjunct populations. Allozyme electrophoresis of 18 loci for the Tono region indicated that 22.2% of the loci are polymorphic (P) with a gene diversity of (h) 0.087. The population of Tsushima has 38.9% polymorphic loci and a gene diversity of 0.149. We could not find any spatial genetic structure of C. retusus in the Tono region by spatial autocorrelation analysis. These results suggest that considerable genetic variation is still present in the Tono region and the influence of the recent habitat fragmentation seems not so large, yet. Because C. retusus is a long-lived tree, the population history may hinder the effect of the fragmentation at present. The genetic diversity (GST) between the populations of the Tono and Tsushima is 0.280, showing the genetic differentiation between these two regions has occurred.

The spatial genetic structure within a population of an endangered plant, Cerastium fischerianum var. molle, was examined using spatial autocorrelation analysis. All individuals in 22 × 15 m area in a population were mapped and genotyped by three allozyme loci. Many spatial autocorrelation indices (Moran's I) were significantly larger than the expected values and their averages across three marker loci examined were positive in the short distance classes and the opposite pattern was shown in the larger distance classes, suggesting that individuals located nearly each other tend to have similar genotypes and pairs of individuals located far from each other tend to have different genotypes. Because the marker loci are considered to be neutral and the linkage disequilibria were not detected, the spatial genetic structure observed in this study seems to result from the restricted gene flow because of predominant selfing of C. fischerianum var. molle.

PREMISE OF THE STUDY: We posed two hypotheses for broad scenarios of postglacial recolonization of Korea by the warm-temperate vegetation: (1) that extant Korean populations are derived from a single refugium, or (2) that they are derived from multiple refugia. We chose a homosporous fern typical of East Asian warm-temperate vegetation, Selliguea hastata, to test which of the two scenarios is more likely and to check whether Japan contained putative glacial refugia. METHODS: Using 16 allozyme loci, we obtained genotypes of 756 individuals from 20 populations, representative of the whole distribution area in Korea (including Jeju Island), Japan, and Taiwan. We assessed genetic variability within and among populations, Wright's F-statistics, and conducted analysis of molecular variance, model-based Bayesian clustering, and bottleneck tests. KEY RESULTS: We found no allozyme variation within populations of S. hastata in mainland Korea, whereas genetic polymorphism was detected for populations from Jeju Island, Japan (in particular a population from southeastern Shikoku), and Taiwan. The levels of inbreeding within populations were high, consistent with the potential of S. hastata for intragametophytic selfing. CONCLUSIONS: Data on allelic richness together with Bayesian clustering methods suggest a pattern of postglacial recolonization of mainland Korea from a single refugium, probably located either on Jeju Island or in Japan. Jeju Island should merit the highest priority for conservation biogeography, as it played a role as a Quaternary refugium for arctic-alpine, boreal, temperate as well as warm-temperate plants, as suggested here.

Premise of the study: Microsatellite markers were developed for Isodon longitubus to study the natural hybridization of the species and its congeners. Methods and Results: A total of 10 primer sets were developed for I. longitubus. From the initial screening, all of 10 loci were polymorphic with five to 19 alleles per locus in the Mt. Ishizuchi population, whereas nine loci were polymorphic with two to 12 alleles per loci in the Toon population. Although one locus was monomorphic at one population, the observed and expected heterozygosity values estimated from 34 I. longitubus samples ranged from 0.273 to 1.000 and from 0.483 to 0.918, respectively. Six primer sets could amplify all three species examined in this study (I. inflexus, I. japonicus, and I. shikokianus). Conclusions: The 10 microsatellite markers developed here will be useful in analyzing the population genetic structure of I. longitubus and in studying the natural hybridization between Isodon species.

We analyzed variations in chloroplast DNA (cpDNA) in the widespread herbaceous species Hosta sieboldiana and Hosta albomarginata across large portions of their geographic ranges in the Japanese archipelago. Our objective was to compare the phylogeographic histories and phylogeographic structures of the two congeneric species in the Japanese archipelago. The location of the study is Japanese archipelago. We sequenced 1380 bp of noncoding cpDNA from 45 populations of H. sieboldiana (n = 362) and 55 populations of H. albomarginata (n = 436) to assess genetic variations within and among populations across almost the entire distributions of the species in Japan. Extant patterns of geographic structure were analyzed using statistical parsimony networks and spatial analysis of molecular variance (SAMOVA). We also used Monmonier's algorithm to detect genetic barriers between regions. Relationships between the populations were examined using a neighbor-joining (NJ) method. Four haplotypes were found for H. sieboldiana, whereas eight haplotypes were identified for H. albomarginata. Total genetic haplotype diversity (hT) and within-population haplotype diversity (hS) for H. sieboldiana were 0.352 and 0.040, respectively, while the values for H. albomarginata were 0.529 and 0.085, respectively. The population differentiations (GST) for H. sieboldiana and H. albomarginata were 0.839 and 0.886, respectively. The SAMOVA analysis revealed two clusters in H. sieboldiana and four clusters in H. albomarginata. Differentiations between and among the clusters were supported by the BARRIER analysis and the NJ tree. We detected differences in the population genetic structure between the two species. We found that H. sieboldiana had lower haplotype diversity than H. albomarginata. These results may be partially explained by the difference in ecological habitats and geographic distributions between the species. Hosta albomarginata is more widely distributed than H. sieboldiana in East Asia including Russia, and this large distribution range would enable more chances to intraspecific gene flow.

We sampled 17 populations of a rare autotetraploid Aster kantoensis (Asteraceae) from three river systems located in central Japan, and studied them for allelic variation at 22 enzyme loci. There was no significant correlation between the actual population size and three genetic diversity parameters, suggesting that the effective population size was very small even for the large populations, i.e., even large populations may still have a high probability of being of recent origin and remain influenced by the founder effect. Compared to other autotetraploid species, the total genetic variation of A. kantoensis is small. The number of alleles and gene diversity of a population were not significantly different among the river systems, although the percentage of polymorphic loci was different. Genetic differentiation among river systems was larger than between populations within the river systems, thereby indicating that gene flow between river systems is small, especially between the Kinu River system and Tama or Sagami River systems.

To examine the response of pollinating bees to size and sexual phases of flowers, we constructed an artificial population ofCampanula having large flower variation and presented it to potentially pollinating bees in nurseries. The pollinating bee groups (halictid, megachilid and bumble bees) responded differentially to both the flower size and to the sexual phases of the flowers. Whereas visitation rate of megachilid bees increased with the flower size, those of halictid bees and bumble bees did not show particular trends; for example, bumble bees visited almost all of the flowers consistently. Visitation frequencies to male-and female-phased flowers were significantly different between megachilids at Tokyo and halictids. This study indicates that pollinator attraction could not solely explain the evolution of the flower size inCampanula, and that other factors such as pollen transfer efficiency, should be considered.

Weigela coraeensis var. coraeensis is a deciduous shrub species distributed in Japan on the mainland, Honshu, whereas its variety W. coraeensis var. fragrans is endemic to the Izu Islands located south of Honshu. We isolated eight polymorphic microsatellite loci from the species and characterized these loci for 20 individuals from a population in Honshu. The number of alleles per locus ranged from 7 to 15 and the observed and expected heterozygosities ranged from 0.60 to 0.90 and from 0.65 to 0.90, respectively. These eight polymorphic microsatellites will be useful for examining intraspecific genetic differentiation in W. coraeensis.

The spatial structure of allele frequencies for three polymorphic enzyme loci was examined in a gynodioecious population of Chionographis japonica var. kurohimensis. Spatial autocorrelation indices (Moran's I) were significantly larger in the shortest distance class than expected value, indicating that genetic similarity is shared among near-neighbor individuals. Because spatial patterns of the three loci examined showed a similar pattern, and significant linkage disequilibrium was not found among pairwise combinations of these loci, the population substructure detected in this study probably resulted from restricted gene flow because of predominant selfing in the hermaphrodites.

Intrapopulation and interpopulation variations in floral sex ratio in hermaphrodites of gynodioeciousChionographis japonica var.kurohimensis (Liliaceae) were examined. The relative ratio of male flowers to total flowers (male and perfect flowers) decreased with plant size, suggesting size-dependent gender modification. The relative ratio of male flowers per population-basis is negatively correlated with the mean number of perfect flowers. Since the number of perfect flowers proportionally increased with plant size, populations showing low maleness consist of relatively bigger plants and are considered to be in high-quality environment. On the other hand, the relative ratio of male flowers per population basis is independent of female frequency in the population. Plasticity in gender expression probably plays an important role of maintenance of gynodioecy inC. japonica var.kurohimensis.

Inter- and intrapopulation pollen-mediated gene flow was estimated in a population of Cynanchum grandifolium var. nikoense by paternity analysis of all the seeds set in 1990. By the simple exclusion procedure, the rate of genes migrating by pollen into this population was estimated to be 0.083; this value is lower than that estimated in most previous studies. Frequency distributions of intrapopulation pollen-mediated gene flow distances estimated by different methods of paternity analysis were platykurtic, and the estimated distances of realized gene flow were smaller than the interplant distance.

The selfing rate of hermaphrodites, inbreeding depression, and relative fecundity of females compared to hermaphrodites were estimated for a gynodioecious population of Chionographis japonica var. kurohimensis to test the models for the evolution of gynodioecy. In spite of the high level of selfing of hermaphrodites, significant amounts of inbreeding depression were found in seed germination and seedling growth. In addition, females produced more fertilized ovules than hermaphrodites. Nevertheless, the conditions theoretically required for maintaining gynodioecy were not satisfied even if the combined effect of these two factors was considered. Additional causes for the evolution of gynodioecy, including biparental inbreeding, are discussed. In some populations of flowering plants, females coexist with hermaphrodites, a condition known as gynodioecy. Because females reproduce sexually only by seeds, they must have some advantage to stably coexist with hermaphrodites. What these advantages are remains an open question in plant evolutionary biology. A number of theoretical works consider the basis for the evolution of gynodioecy. Lloyd (1975, 1976), Charlesworth and Charlesworth (1978), and Charlesworth and Ganders (1979) showed by evolutionarily stable strategy models that gynodioecy can evolve if the product of selfing rate of hermaphrodites (s) and inbreeding depression (d) is over 0.5 when nuclear genes control male sterility. Because viability of selfed progeny is known to be lower

カワラノギクの保全生物学的研究の一環として, 種子の段階の動態について検討した。多摩川で最大の生育地である秋川市草花地区の7つの個体群について, 種子捕食者であるツツミノガ属の一種の幼虫による食害を考慮して, 1991年12月 (一部は翌年12月) に開花個体密度, 個体当りの頭果数などから有効なシードレイン (単位面積当りの種子生産量) を調査した。最大の個体群において, 埋土種子の数の変化を1992年と翌年に調べた。有効なシードレインは1, 200～11,000粒/m2であった。発芽時期の後には埋土種子は少数しか残っていなかったので, 埋土種子集団の個体群の維持に対する貢献は小さいことが判明した。

Approximately one-fourth of Japan's native plant species are threatened with extinction. To conserve these species, it is critical to evaluate genetic diversity at species-level and population-level. Some factors, including population size and geographic distribution, are known to influence the population genetic diversity of wild plant species. This article briefly reviews the population genetic studies that have been conducted on wild threatened plants in Japan. A large population size or wide geographic distribution does not always lead to large genetic diversity, suggesting that historical factors such as speciation processes and population expansion often play more important roles in determining genetic diversity than the number of remnant individuals. The mating system of a species also affects genetic diversity; predominantly selfing species tend to have smaller genetic diversity than outcrossing congeners. Another issue of concern in the conservation genetics of wild plants in Japan is the genetic diversity of insular endemics, because Japan consists of many islands, and the insular flora contains many endemic and threatened species. Previous studies on endemic plants on the Bonin and the Ryukyu Islands are reviewed. Compared to the cases of the Bonin Islands or other oceanic islands, there is much larger genetic diversity in plants endemic to the Ryukyu Islands. This difference is probably the result of the differences in the geological history of these islands.

To examine the evolution of gynodioecy under biparental inbreeding, we developed a new theoretical model and tested it by estimating paramaeters incorporated in the model in a gynodioecious population ofChionographis japonica var.kurohimensis. The model and the results showed biparental inbreeding affects the evolution of gynodioecy, but the effect is very small.