A new perspective on the evolution of "Kawara" roof tiles in Ryukyu: A multidisciplinary non-destructive analysis of roof tile transition at Shuri Castle, Ryukyu Islands, Japan.

A unique historical architecture was created at Shuri Castle (Shuri-jo) in the Ryukyu Islands by its "Kawara" roof tiles. After the 13th and 14th centuries, Kawara tiles were introduced to the Ryukyu Islands from several regions, including China, Korea, and mainland Japan, and evolved shapes and patterns that are unique to this island region. However, the transition of some internal features, such as the chemical components and microstructure, had not been analyzed. This study used a multi-faceted approach for such internal data and non-destructive quantitative methods to propose a new perspective on the evolution of historical Ryukyuan Kawara. We analyzed two styles of Ryukyuan Kawara from the 13th to 15th centuries and found that the material processing and firing conditions of the two styles were very similar, even though it had been suggested that they had different origins. A quantitative analysis of tiles from the 16th to 19th centuries revealed a transition in color tone to red, leading to the modern traditional Ryukyuan tiles; traces of changes in firing conditions were also found along with this transition. Finally, the study revealed that the evolution of Ryukyuan Kawara consisted of changed factors, e.g. surface color, and unchanged factors, e.g. paste density. Previous archaeological studies mainly focused on changing external characteristics, such as form and pattern; however, our analysis showed that the internal features changed, while the elemental composition and paste density remained constant from the appearance of the roof tiles until the 19th century. We propose that this is related to different responses of individual factors to external stressors, such as the social context, which may be common to other archaeological artifacts as well. Our study provides a new perspective on the evolution of Ryukyuan Kawara and presents a different discussion of and methods for the chronological study of material culture.

Mesozoic origin and 'out-of-India' radiation of ricefishes (Adrianichthyidae).

The Indian subcontinent has an origin geologically different from Eurasia, but many terrestrial animal and plant species on it have congeneric or sister species in other parts of Asia, especially in the Southeast. This faunal and floral similarity between India and Southeast Asia is explained by either of the two biogeographic scenarios, 'into-India' or 'out-of-India'. Phylogenies based on complete mitochondrial genomes and five nuclear genes were undertaken for ricefishes (Adrianichthyidae) to examine which of these two biogeographic scenarios fits better. We found that Oryzias setnai, the only adrianichthyid distributed in and endemic to the Western Ghats, a mountain range running parallel to the western coast of the Indian subcontinent, is sister to all other adrianichthyids from eastern India and Southeast-East Asia. Divergence time estimates and ancestral area reconstructions reveal that this western Indian species diverged in the late Mesozoic during the northward drift of the Indian subcontinent. These findings indicate that adrianichthyids dispersed eastward 'out-of-India' after the collision of the Indian subcontinent with Eurasia, and subsequently diversified in Southeast-East Asia. A review of geographic distributions of 'out-of-India' taxa reveals that they may have largely fuelled or modified the biodiversity of Eurasia.

Unraveling a 70-year-old taxonomic puzzle: redefining the genus ikedosoma (annelida: echiura) on the basis of morphological and molecular analyses.

After a long-standing taxonomic confusion, the echiurid genus Ikedosoma Bock, 1942 , endemic to Japan and surroundings, is redefined on the basis of morphological and molecular analyses of many new Japanese materials and some museum specimens. The re-examination of a syntype of I. elegans ( Ikeda, 1904 ), the type species of the genus, first revealed that its oblique muscle layer is continuous throughout and never fasciculate between longitudinal muscle bands, unlike those described in the definitions that have prevailed for ca. 70 years, making this genus indistinguishable from Listriolobus Spengel, 1912 . Two Japanese species of Ikedosoma, I. elegans and I. gogoshimense (Ikeda, 1904), which were thus redefined, had also been poorly defined in the past to the point of being nearly indistinguishable from each other, largely due to incomplete descriptions and poor collections. Molecular phylogenetic analyses using 18S and 28S ribosomal RNA, histone H3, and cytochrome c oxidase subunit I (COI) genes clearly confirmed the distinction between these two species, their monophyletic origin, and their distinction from L. sorbillans (Lampert, 1883). The genus Ikedosoma thus validated is morphologically distinguishable from Listriolobus by the absence of a rectal caecum. Ikedosoma elegans and I. gogoshimense also differ in the disposition of gonoduct pairs. The third known species, I. qingdaoense Li, Wang and Zhou, 1994 , from Qingdao, North China, lacks information on oblique muscle layers, which makes even its generic affiliation uncertain.

Biogeography and evolution of the Carassius auratus-complex in East Asia.

BACKGROUND: Carassius auratus is a primary freshwater fish with bisexual diploid and unisexual gynogenetic triploid lineages. It is distributed widely in Eurasia and is especially common in East Asia. Although several genetic studies have been conducted on C. auratus, they have not provided clear phylogenetic and evolutionary descriptions of this fish, probably due to selection bias in sampling sites and the DNA regions analysed. As the first step in clarifying the evolutionary entity of the world's Carassius fishes, we attempted to clarify the phylogeny of C. auratus populations distributed in East Asia. RESULTS: We conducted a detailed analysis of a large dataset of mitochondrial gene sequences [CR, 323 bp, 672 sequences (528 sequenced + 144 downloaded); CR + ND4 + ND5 + cyt b, 4669 bp in total, 53 sequences] obtained from C. auratus in East Asia. Our phylogeographic analysis revealed two superlineages, one distributed mainly among the Japanese main islands and the other in various regions in and around the Eurasian continent, including the Ryukyus and Taiwan. The two superlineages include seven lineages with high regional specificity that are composed of endemic populations indigenous to each region. The divergence time of the seven lineages was estimated to be 0.2 million years ago (Mya) by a fossil-based method and 1.0-1.9 Mya by the molecular clock method. The antiquity and endemism of these lineages suggest that they are native to their respective regions, although some seem to have been affected by the artificial introduction of C. auratus belonging to other lineages. Triploids of C. auratus did not form a monophyletic lineage but were clustered mostly with sympatric diploids. CONCLUSIONS: The results of the present study revealed the existence of two superlineages of C. auratus in East Asia that include seven lineages endemic to each of the seven regions examined. The lack of substantial genetic separation between triploids and diploids indicates that triploids are not composed of a single independent lineage. The ancient origins and evolutionary uniqueness of the seven lineages warrant their conservation. An overall phylogenetic framework obtained from the present study will be of use for estimating the phylogenetic relationships of Carassius fishes on the Eurasian continent.

Phylogenetic position of a whale-fall lancelet (Cephalochordata) inferred from whole mitochondrial genome sequences.

BACKGROUND: The lancelet Asymmetron inferum (subphylum Cephalochordata) was recently discovered on the ocean floor off the southwest coast of Japan at a depth of 229 m, in an anaerobic and sulfide-rich environment caused by decomposing bodies of the sperm whale Physeter macrocephalus. This deep sulfide-rich habitat of A. inferum is unique among the lancelets. The distinguishing adaptation of this species to such an extraordinary habitat can be considered in a phylogenetic framework. As the first step of reconstruction of the evolutionary processes in this species, we investigated its phylogenetic position based on 11 whole mitochondrial genome sequences including the newly determined ones of the whale-fall lancelet A. inferum and two coral-reef congeners. RESULTS: Our phylogenetic analyses showed that extant lancelets are clustered into two major clades, the Asymmetron clade and the Epigonichthys + Branchiostoma clade. A. inferum was in the former and placed in the sister group to A. lucayanum complex. The divergence time between A. inferum and A. lucayanum complex was estimated to be 115 Mya using the penalized likelihood (PL) method or 97 Mya using the nonparametric rate smoothing (NPRS) method (the middle Cretaceous). These are far older than the first appearance of large whales (the middle Eocene, 40 Mya). We also discovered that A. inferum mitogenome (mitochondrial genome) has been subjected to large-scale gene rearrangements, one feature of rearrangements being unique among the lancelets and two features shared with A. lucayanum complex. CONCLUSION: Our study supports the monophyly of genus Asymmetron assumed on the basis of the morphological characters. Furthermore, the features of the A. inferum mitogenome expand our knowledge of variation within cephalochordate mitogenomes, adding a new case of transposition and inversion of the trnQ gene. Our divergence time estimation suggests that A. inferum remained a member of the Mesozoic and the early Cenozoic large vertebrate-fall communities before shifting to become a whale-fall specialist.

DNA sequences identify numerous cryptic species of the vertebrate: a lesson from the gobioid fish Schindleria.

Schindleria (Gobioidei, Schindleriidae), believed to include one of the smallest and youngest reproducing vertebrates, is broadly distributed in the Indo-Pacific Oceans, inhabiting coral reef lagoons. They are all characterized by a reduced larval-like form, such as a slender translucent and scaleless body. The three nominal species recognized in the genus to date have been distinguished by only combination of dorsal and anal fin-ray counts, and the existence of some undescribed species has been suggested in Schindleria; thus a total picture of species composition of the genus is poorly known. Towards the disclosure of diversity of Schindleria, a molecular phylogenetic analysis using partial mitochondrial 16S rRNA sequences was conducted for specimens from the Ryukyu and Ogasawara Islands, Japan. This analysis showed clearly that as many as 21 genetically distinguishable species occurred within the geographical areas. The degree of species crypticness of "S. praematura" [15.0=15 (new cryptic species + known species)/1 (known species)] is higher than the values of well-known animal examples, such as the pan-mesopelagic bristlemouth fish Cyclothone alba (5.0) and the South American skipper butterfly Astraptes fulgerator (10.0). This discovery of many cryptic species in Schindleria suggests that the use of DNA sequences is necessary for species identification of such morphologically conserved taxa. Because molecular analyses should increase the number of hitherto unnamed and pseudonymous species, especially in tropical areas, it is proposed that DNA-based designation is necessary for such taxa in order to compile the full "species lists", although there is presently no consensus for the inclusion of DNA sequencing data in the formal descriptions of new species.