A DNA barcode reference library for the native woody seed plants of Japan.

DNA barcode databases are increasingly available for a range of organisms, facilitating the wide application of DNA barcode-based studies. Here we announce the development of a comprehensive DNA barcode reference library of Japanese native woody seed plants representing 43 orders, 99 families, 303 genera and 834 species, and comprising 77.3% of the genera and 72.2% of the species of native woody seed plants in Japan. A total of 6216 plant specimens were collected from 223 sites across the subtropical, temperate, boreal and alpine biomes in Japan with most species represented by multiple accessions. This reference library utilized three chloroplast DNA regions (rbcL, trnH-psbA and matK) and consists of 14,403 barcode sequences. Individual regions varied in their identification rates, with species-level and genus-level rates for rbcL, trnH-psbA and matK based on blast being 57.4%/96.2%, 78.5%/99.1% and 67.8%/98.1%, respectively. Identification rates were higher using region combinations, with total species-level rates for two region combinations (rbcL & trnH-psbA, rbcL & matK and trnH-psbA & matK) ranging between 90.6% and 95.8%, and for all three regions being equal to 98.6%. Genus-level identification rates were even higher, ranging between 99.7% and 100% for two region combinations and being 100% for the three regions. These results indicate that this DNA barcode reference library is an effective resource for investigations of native woody seed plants in Japan using DNA barcodes and provides a useful template for the development of libraries for other components of the Japanese flora.

Complete chloroplast genome of Cerasus kumanoensis (Rosaceae), a wild flowering cherry endemic to Kii Peninsula, Japan.

Cerasus kumanoensis is a recently described wild cherry species from the Kii Peninsula, Japan. Here we determined the first complete chloroplast genome of C. kumanoensis using genome skimming approach. The cp genome was 157,898 bp long, with a large single-copy region (LSC) of 85,926 bp and a small single-copy region (SSC) of 19,070 bp separated by a pair of inverted repeats (IRs) of 26,451 bp. It encodes 129 genes, including 84 protein-coding genes, 37 tRNA genes, and 8 ribosomal RNA genes. Besides, we reconstructed the phylogeny of Prunus using maximum likelihood (ML) method, including our data and previously reported cp genomes of related taxa. The phylogenetic analysis indicated that C. kumanoensis is close related with a group including Prunus takesimensis and P. speciosa.

Complete chloroplast genome of Prunus itosakura (Rosaceae), a wild flowering cherry from Japan.

Prunus itosakura is a flowering tree species with high ornamental and economic values. We determined the first complete chloroplast genome of P. itosakura using genome skimming approach. The cp genome was 157,813 bp long, with a large single-copy region (LSC) of 85,931 bp and a small single-copy region (SSC) of 19,120 bp separated by a pair of inverted repeats (IRs) of 26,381 bp. It encodes 129 genes, including 84 protein-coding genes, 37 tRNA genes, and 8 ribosomal RNA genes. We also reconstructed the phylogeny of Prunus sensu lato using maximum likelihood (ML) method, including our data and previously reported cp genomes of related taxa. The phylogenetic analysis indicated that P. itosakura is closely related with Prunus subhirtella var. subhirtella.

Complete chloroplast genome of the wild Japanese Mountain cherry (Prunus jamasakura, Rosaceae).

Prunus jamasakura is a species of Prunus native to eastern Asia. We determined the first complete chloroplast genome of Prunus jamasakura using genome skimming approach. The cp genome was 157,905 bp long, with a large single-copy region (LSC) of 85,910 bp and a small single-copy region (SSC) of 19,123 bp separated by a pair of inverted repeats (IRs) of 26,436 bp. It encodes 129 genes, including 84 protein-coding genes, 37 tRNA genes, and 8 ribosomal RNA genes. We also reconstructed the phylogeny of Prunus sensu lato using maximum likelihood (ML) method, including our data and previously reported cp genomes of related taxa. The phylogenetic analysis indicated that P. jamasakura is closely related with Prunus speciosa.

Modeling daily flowering probabilities: expected impact of climate change on Japanese cherry phenology.

Understanding the drivers of phenological events is vital for forecasting species' responses to climate change. We developed flexible Bayesian survival regression models to assess a 29-year, individual-level time series of flowering phenology from four taxa of Japanese cherry trees (Prunus spachiana, Prunus × yedoensis, Prunus jamasakura, and Prunus lannesiana), from the Tama Forest Cherry Preservation Garden in Hachioji, Japan. Our modeling framework used time-varying (chill and heat units) and time-invariant (slope, aspect, and elevation) factors. We found limited differences among taxa in sensitivity to chill, but earlier flowering taxa, such as P. spachiana, were more sensitive to heat than later flowering taxa, such as P. lannesiana. Using an ensemble of three downscaled regional climate models under the A1B emissions scenario, we projected shifts in flowering timing by 2100. Projections suggest that each taxa will flower about 30 days earlier on average by 2100 with 2-6 days greater uncertainty around the species mean flowering date. Dramatic shifts in the flowering times of cherry trees may have implications for economically important cultural festivals in Japan and East Asia. The survival models used here provide a mechanistic modeling approach and are broadly applicable to any time-to-event phenological data, such as plant leafing, bird arrival time, and insect emergence. The ability to explicitly quantify uncertainty, examine phenological responses on a fine time scale, and incorporate conditions leading up to an event may provide future insight into phenologically driven changes in carbon balance and ecological mismatches of plants and pollinators in natural populations and horticultural crops.

Clone identification in Japanese flowering cherry (Prunus subgenus Cerasus) cultivars using nuclear SSR markers.

Numerous cultivars of Japanese flowering cherry (Prunus subgenus Cerasus) are recognized, but in many cases they are difficult to distinguish morphologically. Therefore, we evaluated the clonal status of 215 designated cultivars using 17 SSR markers. More than half the cultivars were morphologically distinct and had unique genotypes. However, 22 cultivars were found to consist of multiple clones, which probably originate from the chance seedlings, suggesting that their unique characteristics have not been maintained through propagation by grafting alone. We also identified 23 groups consisting of two or more cultivars with identical genotypes. Most members of these groups were putatively synonymously related and morphologically identical. However, some of them were probably derived from bud sport mutants and had distinct morphologies. SSR marker analysis provided useful insights into the clonal status of the examined Japanese flowering cherry cultivars and proved to be a useful tool for cultivar characterization.

Use of two population metrics clarifies biodiversity dynamics in large-scale monitoring: the case of trees in Japanese old-growth forests: the need for multiple population metrics in large-scale monitoring.

Many indicators/indices provide information on whether the 2010 biodiversity target of reducing declines in biodiversity have been achieved. The strengths and limitations of the various measures used to assess the success of such measures are now being discussed. Biodiversity dynamics are often evaluated by a single biological population metric, such as the abundance of each species. Here we examined tree population dynamics of 52 families (192 species) at 11 research sites (three vegetation zones) of Japanese old-growth forests using two population metrics: number of stems and basal area. We calculated indices that track the rate of change in all species of tree by taking the geometric mean of changes in population metrics between the 1990s and the 2000s at the national level and at the levels of the vegetation zone and family. We specifically focused on whether indices based on these two metrics behaved similarly. The indices showed that (1) the number of stems declined, whereas basal area did not change at the national level and (2) the degree of change in the indices varied by vegetation zone and family. These results suggest that Japanese old-growth forests have not degraded and may even be developing in some vegetation zones, and indicate that the use of a single population metric (or indicator/index) may be insufficient to precisely understand the state of biodiversity. It is therefore important to incorporate more metrics into monitoring schemes to overcome the risk of misunderstanding or misrepresenting biodiversity dynamics.

Genetic structure of Cerasus jamasakura, a Japanese flowering cherry, revealed by nuclear SSRs: implications for conservation.

The genetic resources of a particular species of flowering cherry, Cerasus jamasakura, have high conservation priority because of its cultural, ecological and economic value in Japan. Therefore, the genetic structures of 12 natural populations of C. jamasakura were assessed using ten nuclear SSR loci. The population differentiation was relatively low (F (ST), 0.043), reflecting long-distance dispersal of seeds by animals and historical human activities. However, a neighbor-joining tree derived from the acquired data, spatial analysis of molecular variance and STRUCTURE analysis revealed that the populations could be divided into two groups: one located on Kyusyu Island and one on Honshu Island. Genetic diversity parameters such as allelic richness and gene diversity were significantly lower in the Kyushu group than the Honshu group. Furthermore, STRUCTURE analysis revealed that the two lineages were admixed in the western part of Honshu Island. Thus, although the phylogeographical structure of the species and hybridization dynamics among related species need to be evaluated in detail using several marker systems, the Kyusyu Island and Honshu Island populations should be considered as different conservation units, and the islands should be regarded as distinct seed transfer zones for C. jamasakura, especially when rapid assessments are required.

Impact of global warming on a group of related species and their hybrids: cherry tree (Rosaceae) flowering at Mt. Takao, Japan.

Climate change is affecting plant phenology worldwide. Phenological responses vary among species, but it is not clear how responses differ among closely related species. We examined a 25-yr record (1981-2005) of flowering times for 97 trees, representing 17 species and hybrids of cherry (Cerasus sp. or Prunus sp.) grown at Mt. Takao, in Tokyo, Japan. The cherry trees flowered earlier over time, by an average of 5.5 d over the 25-yr study. Earlier flowering was explained largely by a 1.8°C increase in February-March mean monthly temperatures. Most species and hybrids flowered 3-5 d earlier for each 1°C increase in temperature, but early-flowering taxa flowered as much as 9 d earlier for each 1°C increase in temperature. Flowering durations and differences in flowering times among species were greater in warm years than in cold years. Species and individual trees also flowered longer in warm years. These results show that the flowering times of closely related species may change similarly in response to climate change, but that early-flowering species may diverge from the overall trend in a predictable way. Such changes in flowering may affect gene flow and pollination as the length of the flowering season increases.