Hiroshi Kudoh

Evolution and Control of Imprinted FWA Genes in the Genus Arabidopsis

A central question in genomic imprinting is how a specific sequence is recognized as the target for epigenetic marking. In both mammals and plants, imprinted genes are often associated with tandem repeats and transposon-related sequences, but the role of these elements in epigenetic gene silencing remains elusive. FWA is an imprinted gene in Arabidopsis thaliana expressed specifically in the female gametophyte and endosperm. Tissue-specific and imprinted expression of FWA depends on DNA methylation in the FWA promoter, which is comprised of two direct repeats containing a sequence related to a SINE retroelement. Methylation of this element causes epigenetic silencing, but it is not known whether the methylation is targeted to the SINE-related sequence itself or the direct repeat structure is also necessary. Here we show that the repeat structure in the FWA promoter is highly diverse in species within the genus Arabidopsis. Four independent tandem repeat formation events were found in three closely related species. Another related species, A. halleri, did not have a tandem repeat in the FWA promoter. Unexpectedly, even in this species, FWA expression was imprinted and the FWA promoter was methylated. In addition, our expression analysis of FWA gene in vegetative tissues revealed high frequency of intra-specific variation in the expression level. In conclusion, we show that the tandem repeat structure is dispensable for the epigenetic silencing of the FWA gene. Rather, SINE-related sequence is sufficient for imprinting, vegetative silencing, and targeting of DNA methylation. Frequent independent tandem repeat formation events in the FWA promoter led us to propose that they may be a consequence, rather than cause, of the epigenetic control. The possible significance of epigenetic variation in reproductive strategies during evolution is also discussed.

The effect of petal size manipulation on pollinator/seed-predator mediated female reproductive success of Hibiscus moscheutos

Abstract The effects of petal-size manipulations on the behavior of pollinators and pollen/seed predators, and on pollen removal and deposition, were studied in Hibiscus moscheutos (Malvaceae) populations. The ultimate effects on the female reproductive success of flowers, such as fruit set, seed predation rate, and final seed set were also measured. We applied three levels of petal removal (100%, 50%, and 0% size reduction in radius) to flowers in natural populations. Two pollinators (Bombus pennsylvanicus and Ptilothrix bombiformis) ignored flowers without petals, suggesting that pollinators use petals as a visual cue to locate flowers. Consequently, 100% petal removal reduced female reproductive success considerably, mainly through a higher rate of fruit abortion due to failure of pollen deposition on stigmas. No significant differences between the 50% petal removal treatment and uncut control were detected in any components of female success examined. The results, therefore, suggest that differences in petal size have little influence on female reproductive success of Hibiscus flowers at our study site. Final seed set varied considerably depending on the larval densities of two coleopteran seed predators (Althaeus hibisci and Conotrachelus fissunguis). A. hibisci responded to petal size, and a higher density of adults was found in flowers in which petal size had not been reduced. Because Althaeus feed on pollen as adults and no effect of petal size on seed predation was detected, the preference of Althaeus for larger flowers may represent a foraging strategy for adult beetles and may exert counteracting selection pressure on petal size through male reproductive success of flowers.

Does invasion involve alternation of germination requirements? A comparative study between native and introduced strains of an annual Brassicaceae, Cardamine hirsuta

Cardamine hirsuta is a European annual weed that has been naturalized in Japan. Although the species is a widespread weed in Europe, its introduction to the Japanese Islands occurred recently. We hypothesized that the introduction of Cardamine hirsuta required adaptation that has delayed its spread in Japan. The aim of this study is to determine whether there is a difference in temperature requirements for seed germination between Japanese and European strains of Cardamine hirsuta. We compared temperature requirements for seed germination, because it is known to be a critical determinant of phenology and thus is expected to be important in the success of introduction into different climates. Seeds of six and five strains from Japan and Europe, respectively, were used in seed germination experiments under five different temperature regimes. Japanese strains generally showed stronger initial seed dormancy and were characterized by suppressed germination in higher temperature regimes. European strains showed variable patterns of temperature-dependent seed germination. It turned out that these temperature-dependent dormancies are mediated by abscisic acid (ABA), because dormant seeds germinated under the presence of an inhibitor of ABA synthesis. Seed germination characteristics shared by Japanese strains presumably enhance long transport and autumn germination, while these characteristics are not necessarily common among European strains. The results are supportive for the idea that adaptation has altered germination characteristics of Cardamine hirsuta through the introduction process into Japan.

Origin and diversification of Hibiscus glaber , species endemic to the oceanic Bonin Islands, revealed by chloroplast DNA polymorphism

Two woody Hibiscus species co‐occur in the Bonin Islands of the northwestern Pacific Ocean: Hibiscus glaber Matsum. is endemic to the islands, and its putative ancestral species, Hibiscus tiliaceus L., is widely distributed in coastal areas of the tropics and subtropics. To infer isolating mechanisms that led to speciation of H. glaber and the processes that resulted in co‐occurrence of the two closely related species on the Bonin Islands, we conducted molecular phylogenetic analyses on chloroplast DNA (cpDNA) sequences. Materials collected from a wide area of the Pacific and Indian Oceans were used, and two closely related species, Hibiscus hamabo Siebold Zucc. and Hibiscus macrophyllus Roxb., were also included in the analyses. The constructed tree suggested that H. glaber has been derived from H. tiliaceus, and that most of the modern Bonin populations of H. tiliaceus did not share most recent ancestry with H. glaber. Geographic isolation appears to be the most important mechanism in the speciation of H. glaber. The co‐occurrence of the two species can be attributed to multiple migrations of different lineages into the islands. While a wide and overlapping geographical distribution of haplotypes was found in H. tiliaceus, localized geographical distribution of haplotypes was detected in H. glaber. It is hypothesized that a shift to inland habitats may have affected the mode of seed dispersal from ocean currents to gravity and hence resulted in geographical structuring of H. glaber haplotypes.

Breeding system of the annual Cruciferae, Arabidopsis kamchatica subsp. kawasakiana

The breeding system of an annual Cruciferae, Arabidopsis kamchatica subsp. kawasakiana, was studied in three natural populations. We applied four experimental treatments, open pollination, bagging, emasculationxa0+xa0bagging, and emasculationxa0+xa0hand-pollinationxa0+xa0bagging. None of the emasculated flowers with bags produced fruits but we observed high fruit sets in the other three treatments. The results confirmed that A. kamchatica subsp. kawasakiana is a self-compatible, non-apomictic species that can produce seeds through auto-pollination. Considering the life cycle as an annual, increased reproductive assurance through auto-pollination should be critical for the maintenance of populations of A. kamchatica subsp. kawasakiana.

Patch structure and ramet demography of the clonal tree, Asimina triloba, under gap and closed canopy

Clonal understory trees develop into patches of interconnected and genetically identical ramets that have the potential to persist for decades or centuries. These patches develop beneath forest canopies that are structurally heterogeneous in space and time. Canopy heterogeneity, in turn, is responsible for the highly variable understory light environment that is typically associated with deciduous forests. We investigated what aspects of patch structure (density, size structure, and reproductive frequency of ramets) of the clonal understory tree, Asimina triloba, were correlated with forest canopy conditions. Specifically, we compared A. triloba patches located beneath closed canopies and canopy gaps. We also conducted a three-year demographic study of individual ramets within patches distributed across a light gradient. The closed canopy-gap comparison demonstrated that the patches of A. triloba had a higher frequency of large and flowering ramets in gaps compared to closed-canopy stands, but total ramet density was lower in gaps than in closed canopy stands. In the demographic study, individual ramet growth was positively correlated with light availability, although the pattern was not consistent for all years. Neither ramet recruitment nor mortality was correlated with light conditions. Our results indicate that the structure of A. triloba patches was influenced by canopy condition, but does not necessarily depend on the responses of ramets to current light conditions. The lack of differences in ramet recruitment and mortality under varying canopy conditions is likely to be a primary reason for the long-term expansion and persistence of the patches. The primary benefit of a positive growth response to increasing light is the transition of relatively small ramets into flowering ramets within a short period of time.

The correct interpretation and lectotypification of the name Cardamine fallax (Brassicaceae)

The name Cardamine fallax (O. E. Schulz) Nakai, based on Cardamine flexuosa subsp. fallax O. E. Schulz, is lectotypified by the specimen originating from Japan (Mama-mura, Shimosa) in accordance with the original description and with the current use of the name by the majority of Japanese and Korean authors. Contrary to the treatment in the recent editions of the Flora of China and Flora of Japan, hexaploid C. fallax is considered here as a taxon different from diploid C. parviflora L. The main morphological difference between these two species is in the shape of cauline leaves. Those of C. parviflora are pinnatisect (lower ones seldom pinnate), with oblanceolate to linear, entire or almost entire segments or leaflets, and those of C. fallax are pinnate, usually with petiolulate, lobate, pinnatipartite to pinnatisect leaflets. The distribution area of C. fallax includes Japan, Korea and Eastern China.