Double migration of the endangered Tricyrtis formosana (Liliaceae) in Japan.

The Ryukyu Islands of Japan are a biodiversity hotspot due to geographical and historical factors. Tricyrtis formosana is a perennial herbaceous plant that commonly found in Taiwan. But only a few populations have been identified in a limited habitat on Iriomote Island, while populations of unknown origin occur near human settlements in an area on the main island of Okinawa. To better understand these populations of the phylogenetic uniqueness and intrinsic vulnerability, we conducted comparative analyses including (1) phylogeny and population structure with MIG-seq data, (2) photosynthesis-related traits of plants grown under common conditions and (3) transcriptome analysis to detect deleterious variations. Results revealed that T. formosana was split into two clades by the congeners and that Iriomote and Okinawa populations independently derived from ancestral Taiwanese populations in each clade. Photosynthetic efficiency was lowest in the Iriomote population, followed by Okinawa and Taiwan. Transcriptome analysis showed that the Iriomote population accumulated more deleterious variations, suggesting intrinsic vulnerability. These results indicate that each T. formosana population in Japan is phylogenetically unique and has been independently dispersed from Taiwan, and that the Iriomote population presents a high conservation difficulty with a unique photosynthesis-related characteristic and a larger amount of deleterious variations.

Genomic insights into rapid speciation within the world's largest tree genus Syzygium.

Species radiations, despite immense phenotypic variation, can be difficult to resolve phylogenetically when genetic change poorly matches the rapidity of diversification. Genomic potential furnished by palaeopolyploidy, and relative roles for adaptation, random drift and hybridisation in the apportionment of genetic variation, remain poorly understood factors. Here, we study these aspects in a model radiation, Syzygium, the most species-rich tree genus worldwide. Genomes of 182 distinct species and 58 unidentified taxa are compared against a chromosome-level reference genome of the sea apple, Syzygium grande. We show that while Syzygium shares an ancient genome doubling event with other Myrtales, little evidence exists for recent polyploidy events. Phylogenomics confirms that Syzygium originated in Australia-New Guinea and diversified in multiple migrations, eastward to the Pacific and westward to India and Africa, in bursts of speciation visible as poorly resolved branches on phylogenies. Furthermore, some sublineages demonstrate genomic clines that recapitulate cladogenetic events, suggesting that stepwise geographic speciation, a neutral process, has been important in Syzygium diversification.

Multiple origins of two Ochrosia (Apocynaceae) species endemic to the Bonin (Ogasawara) Islands.

The genus, Ochrosia, is widely distributed from the West Indian Ocean throughout tropical Asia to the Middle Southern Pacific region. Ochrosia comprises many island-endemic species, suggesting that long-distance dispersal and isolation after migration are key factors for clarifying the diversification process. However, the phylogeny and biogeography of endemic Ochrosia species have not been evaluated well due to the difficulty of adequate sampling from the entire distribution range of the genus. In this study, we focused on two Ochrosia species endemic to the Bonin (Ogasawara) Islands in the northwest Pacific. The Bonin Islands are of volcanic origins and consist of two islands groups, the Ogasawara and Volcano Islands groups, approximately 300 km apart. Ochrosia nakaiana is endemic to the Ogasawara Islands group, whereas O. hexandra is endemic to the Volcano Islands group. To elucidate the phylogenetic positions of these two endemic Ochrosia species, we conducted molecular phylogenetic studies with dating and biogeographic analyses including other Ochrosia species. The phylogenetic trees showed that the two endemic species had distinct origins; O. nakaiana was closely related to O. oppositifolia and O. iwasakiana, whereas O. hexandra was related to O. mariannensis. Based on the chloroplast DNA phylogeny, the genus, Ochrosia, divided into two major lineages 36.6 million years ago. Further, the two endemic species of the Bonin Islands were independently derived approximately 1-2 million years ago. Ochrosia nakaiana originated from the Southeast Asia, New Caledonia, or other Pacific Islands, while O. hexandra derived from O. mariannensis in Micronesia. We demonstrated different origins of the two endemic Ochrosia species on the Bonin Islands. This study provided an excellent example of the complex origins and speciation of flora in the oceanic islands.

Comparisons among populations and individuals to evaluate pollen-pistil interaction as a mechanism of reproductive interference in Taraxacum.

Reproductive interference (RI), an interspecific mating interaction that reduces the fitness of at least one of the species involved, can lead to exclusive distributions in closely related species. A hypothesis previously proposed is that RI in plants may occur by ovule usurpation, in which pistils lack interspecific incompatibility and mistakenly accept heterospecific pollen, thereby losing an opportunity for conspecific pollen fertilization. However, few comparative studies have evaluated the consistency of the inferred mechanism within and among individuals and populations. We conducted hand-pollination experiments in six populations of three native Taraxacum species that suffered from different levels of RI from an alien congener, T. officinale, and compared pollen-pistil interactions among populations. We also investigated the interactions for eight individual T. japonicum plants whose response to heterospecific pollen deposition had been previously measured. Our results revealed that pollen tubes often penetrated native ovaries following heterospecific pollination in populations suffering from strong RI, whereas they seldom did in populations suffering from marginal RI. However, the relative frequency of the pollen tube penetration was not significantly related to the strength of alien RI. Not all pistils on an individual plant showed the same pollen receptivity following heterospecific pollination; rather, some accepted and some refused the pollen tubes. The relationship between pollen tube penetration following heterospecific pollination and the strength of the alien RI was also not significant among individuals. Our present results generally support the ovule usurpation hypothesis, but suggest that other factors, such as competition for pollinator services, variation in the effects of heterospecific pollen donors, and condition of the native inflorescences, might also affect the observed RI strength.

 Hydrangeamarunoi (Hydrangeaceae), a new species from Osumi Peninsula, southern Japan.

Hydrangeamarunoi Tagane & S. Fujii, from the Kimotsuki Mountains in the Ohsumi Peninsula, southern Japan, is described and illustrated. It is morphologically similar to H.alternifolia in having three-petaloid calyx lobes in marginal flowers, but is distinguished by the larger stamen number, and longer styles and seeds. Multiplex ISSR genotyping by sequencing (MIG-seq) demonstrated that the new species is monophyletic and closely related to H.amamiohsimensis and H.moellendorffii rather than H.alternifolia.

Habitat partitioning in native Geranium species through reproductive interference.

BACKGROUND AND AIMS: Heterospecific pollen transfer may reduce the fitness of recipient species, a phenomenon known as reproductive interference. A theoretical study has predicted that distributions of species pairs affected by reproductive interference may be syntopic under negligible reproductive interference, sympatric but with partitioning at small spatial scale (i.e. allotopic) under weak interference, or exclusive when reproductive interference is strong. Verifying these predictions is essential for evaluation of the applicability of reproductive interference as a general assembly rule of biological communities. The aim of this study was to test these predictions in two sympatrically distributed wild Geranium species, G. thunbergii and G. wilfordii. METHODS: To measure the effect of reproductive interference, the associations between the relative abundance of the counterpart species and seed set in the focal species, and seed set reduction following mixed pollination, were analysed. The possibility of hybridization with viable offspring was examined by genotyping plants in the field and after mixed pollination. Fertility of putative hybrids was based on their seed set and the proportion of pollen grains with apertural protrusions. A transect study was conducted to examine spatial partitioning, and possible influences of environmental conditions (canopy openness and soil moisture content) on partitioning between the species were analysed. KEY RESULTS: Neither abundance of the counterpart species nor heterospecific pollen deposition significantly affected seed set in the focal species, and hybridization between species was almost symmetrical. Putative hybrids had low fertility. The two species were exclusively distributed at small scale, although environmental conditions were not significantly different between them. CONCLUSIONS: The allotopy of the two species may be maintained by relatively weak reproductive interference through bidirectional hybridization. Re-evaluation of hybridization may allow ongoing or past reproductive interference to be recognized and provide insight into the distributional relationships between the interacting plants.

Comparison of mechanisms of reproductive interference in Taraxacum.

BACKGROUND AND AIMS: Reproductive interference may reduce fitness of either of the involved species, with potentially important ecological and evolutionary consequences. Except for the effect of shared pollinators on reproductive success, however, mechanisms underlying reproductive interference have been little studied, even though the severity of its impact may depend on the specific mechanism. The aim of this study was therefore to explore the mechanisms of reproductive interference between Taraxacum japonicum (native to Japan) and Taraxacum officinale (alien). METHODS: In a field survey, the association between alien species density and seed set in T. japonicum, and whether pollinator behaviour indicated a preference for the alien, were examined. Effects of heterospecific pollen deposition were measured in a series of hand pollination experiments, including mixed pollination experiments in which the order of application of conspecific and heterospecific pollen was varied. Finally, to investigate hybridization frequency, the parentage of seedlings produced following natural, mixed or heterospecific pollination was compared. KEY RESULTS: Alien species density did not negatively affect native seed set, nor did pollinators appear to have a preference for alien flowers. The hand pollination experiments showed that heterospecific pollen deposition adversely affected native seed set, especially when alien pollen was applied before conspecific pollen. No viable hybrids were found following natural pollination, which suggests that hybridization might be a rare event. CONCLUSION: Among the examined mechanisms, heterospecific pollen deposition might have the largest deleterious effect on the native species. This effect is frequency dependent; thus, a positive feedback loop may cause the effect on the population dynamics to increase over time, with the result that the alien might eventually displace the native in a population. Effects of the examined mechanisms on population dynamics should be investigated further to improve understanding of the impact of reproductive interference on the structure of plant communities.

Macrosolen bidoupensis (Loranthaceae), a new species from Bidoup Nui Ba National Park, southern Vietnam.

Macrosolen bidoupensis Tagane & V.S.Dang, sp. nov. (Loranthaceae) is newly described from Bidoup Nui Ba National Park in Lam Dong Province, southern Vietnam. The new species is characterized by small broadly elliptic to circular leaves, sessile to short petioles, slightly cordate to rounded leaf bases, 4-5 pairs of lateral veins and a basally green corolla tube. An illustration, a summary of DNA barcoding of the plastid genes rbcL and matK, and a key to the species of Macrosolen in Vietnam are provided.

Garcinia hopii (Clusiaceae), a new species from Bidoup Nui Ba National Park, southern Vietnam.

A new species, Garcinia hopii H.Toyama & V.S.Dang is described from Bidoup Nui Ba National Park, southern Vietnam. This species is similar to Garcinia hendersoniana Whitmore but differs from that species in having larger leaves, clustered pistillate flowers, a greater number of sterile anthers and a larger stigma of young fruits. A description, preliminary conservation assessment, illustration, photographs and DNA barcodes of the new species are provided, as well as an updated key to Garcinia sect. Hebradendron in Indochina.

A new species of Eustigma (Hamamelidaceae) from Hon Ba Nature Reserve, Vietnam.

A new species of Hamamelidaceae, Eustigma honbaense H.Toyama, Tagane & V.S.Dang, sp. nov., is described from Hon Ba Nature Reserve, Vietnam. This species is similar to Eustigma oblongifolium Gardner & Champ., but differs from it in having entire leaves, longer infructescences, capsules with a longer apical part and seeds with a larger hilum. A description, preliminary conservation assessment, illustration and photographs of the new species are provided, as well as an updated key to the genus Eustigma.

Aporosa tetragona Tagane & V. S. Dang (Phyllanthaceae), a new species from Mt. Hon Ba, Vietnam.

A new species, Aporosa tetragona Tagane & V. S. Dang, sp. nov., is described and illustrated from Mt. Hon Ba located in the Khanh Hoa Province, South Vietnam. This species is characterized by tetragonal pistillate flowers and fruits, which are clearly distinguishable from the other previously known species of the genus. The morphology and phylogeny based on rbcL and matK of this species indicated that the new species belongs to section Appendiculatae Pax & K. Hoffm.

Pollen morphology of the genus Skimmia (Rutaceae) and its taxonomic implications.

Pollen morphology of the genus Skimmia was studied. Of six species of the genus, five species were investigated by light microscopy and scanning electron microscopy. Pollen grains of Skimmia had striate to striate-reticulate sculpture and variable aperture number (3-7), and these characters differed from even the phylogenetically closest relatives such as Dictamnus and Casimiroa which have 3-colporate pollen grains with reticulate exine. In Skimmia laureola, pollen dimorphism is suggested in the aperture number in accordance with the floral system. Pollen grains of Skimmia japonica, a morphologically very variable species, were extensively surveyed covering its whole distribution area. The grains were also striate to striate-reticulate, and the exine sculpture was not different between varieties. However, the aperture number of S. japonica showed a geographical variation ranging from 4 to 6; in Sakhalin, Hokkaido, most part of Honshu and Taiwan, grains with 4-5 apertures were dominant, while in the southern part of Japan, including the Ryukyu Islands and the Izu Islands, those with 5-6 apertures were common.

Karyotypic analysis of Skimmia japonica (Rutaceae) and related species.

A karyotypic analysis of three species of Skimmia (Rutaceae) in East Asia was performed that examined 88 individuals from 53 localities. Chromosome numbers of S. japonica, S. reevesiana and S. arisanensis were 2n=30, 31, 32 (=30+0-2B), 2n=60 and 2n=60, respectively. The chromosome number of S. arisanensis was reported for the first time. All species had a large chromosome pair or quartet (the first pair or quartet) with a median-submedian centromere in the karyotype. In S. japonica the arm ratio of this first pair was considerably variable and showed a geographical pattern. In the northern half of the distribution range, Sakhalin, Hokkaido, Honshu, Shikoku and part of Kyushu, the arm ratio was 1-1.2, while in the southern half, part of Kyushu, Ryukyu and Taiwan, the arm ratio was very variable and ranged from 1.2 to 2.4. In S. japonica the first pair was sometimes rather heteromorphic; however, the heteromorphism was not related to sex of the plant.

Correlation between distyly and ploidy level in Damnacanthus (Rubiaceae).

Somatic chromosomes were observed in 661 individuals of 14 taxa, nine species and five varieties, of Damnacanthus (Rubiaceae). Chromosome numbers are reported for the first time for 13 taxa. Diploid (2n = 22) and tetraploid (2n = 44) counts were obtained. Distyly is reported for the first time for four species, D. angustifolius, D. henryi, D. labordei, and D. officinarum. A strong correlation exists between chromosome number and occurrence of distyly. Regardless of taxa in Damnacanthus, distylous populations are diploid, and monomorphic populations are tetraploid. Flowers of the monomorphic populations observed have a long style and short stamens with few exceptions. Polyploidization may have caused the breakdown of distylous to monomorphic flowers. In D. indicus, leaves from the tetraploid populations tend to be larger than those from the diploid populations. Populations of tetraploid D. indicus were distributed in more northern areas than those of the diploid. Three types of sympatric distribution were found for the varieties of D. indicus in Japan: diploid and tetraploid, two diploids, and two tetraploids. Based on the present chromosome number study, the taxonomy of the varieties of D. indicus should be revised.

Distyly and pollen dimorphism in Damnacanthus (Rubiaceae).

We document distyly in Damnacanthus (Rubiaceae) and investigate pollen morphology in the genus. Within nine taxa in Japan (four species, four varieties and one form), five show distyly. Non-distylous taxa of Damnacanthus always have a long style and short stamens. We examined the pollen morphology of all taxa of Damnacanthus in Japan. The pollen grains of Damnacanthus are 3- to 5-orthocolporate or sometimes loxocolporate. Polymorphic pollen is sometimes observed even within a single flower. Pollen dimorphism is associated with distyly; pollen grains from short-styled (thrum) flowers are significantly larger than those from long-styled (pin) flowers, and the muri are smooth in pin flowers whereas the pollen grains of thrum flowers have minute granules on the top and/or on the sides. In non-distylous taxa, in which the style is longer than stamens, the muri are always provided with minute granules. Interestingly, the pollen grains with granules in the non-distylous taxa contrast with the pollen of the distylous taxa, because granulate pollen grains are found only in thrum flowers. Although Damnacanthus is a stenopalynous genus, the number of colpi is useful for delimiting taxa. Based on palynological characters, we conclude that Damnacanthus should be revised.