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Abstract Background Climatic changes during glacial periods have had a major influence on the recent evolutionary history of living organisms, even in temperate forests on islands, where the land was not covered with ice sheets. We investigated the phylogeographical patterns of the weevil Curculio sikkimensis (Curculionidae), a generalist seed predator of Fagaceae plants living in both deciduous oak and evergreen forests of Japan. Its genetic structure was compared to that of another host-specific seed predator, C. hilgendorfi, inhabiting only evergreen forests. Results We examined 921 bp of mitochondrial DNA for 115 individuals collected from 33 populations of C. sikkimensis from 11 plant species of three genera, Quercus, Lithocarpus, and Castanopsis. An analysis of molecular variance revealed that a large proportion (almost 50%, P < 0.001) of the total genetic variance could be explained by differences between two geographical regions, the southwestern and northeastern parts of the main islands of Japan. In contrast, no significant genetic differentiation of the weevil was observed among vegetation types of their utilized host plant species. The phylogeographical patterns of the generalist and the host-specific seed predator exhibited a congruent genetic boundary in the Chugoku-Shikoku region. Conclusion Our results suggest that geology and historical environment have contributed to shaping the present genetic structure of C. sikkimensis. The geographical patterns of genetic differentiation in the Chugoku-Shikoku region observed in the two types of Fagaceae-associated Curculio in this study have also been observed in several plant species growing in warm and cool temperate zones of Japan. The occurrence of this common pattern suggests that deciduous oak and evergreen forests of Japan survived together, or adjacent to each other, in small refugia during glacial ages, in the southwestern and northeastern parts of the main islands, although these two types of forests are presently distributed in cool and warm temperate zones of Japan, respectively.

Climate changes during glacial periods have had significant effects on the present geographic distribution of plant species. To elucidate the evolutionary history of a plant species with a disjunctive distribution, we investigated the geographic distribution patterns of cpDNA haplotypes in Photinia glabra (Rosaceae) growing in broadleaved evergreen forests in Japan. We examined cpDNA in 42 populations of P. glabra sampled over a geographic range that included Kinki and its surrounding areas and the disjunctive regions in the Amakusa Islands. Both areas had unique cpDNA haplotypes. Moreover, the AMOVA revealed that a large proportion of the total variance (51%, P < 0.001) could be explained by differences among these regions. These results suggest a past fragmentation of this plant species into two separate refugia: southwestern Kyushu and Kinki, including the surrounding area, during the Quaternary glacial periods. A particularly interesting result was that in the southern disjunct distribution in the Amakusa Islands, the genetic subdivision ([[PHI].sub.CT] = 1.00, P < 0.001) appears to lie between the populations from nearly contiguous islands located across a fairway only approximately 80 to 150 m in width. Key words: biogeography; broadleaved evergreen forest; chloroplast DNA; glacial refugia; intraspecific variation; Japan; Photinia glabra; phylogeography.

The broadleaved evergreen forests of the East Asian warm temperate zone are characterised by their high biodiversity and endemism, and there is therefore a need to extend our understanding of its genetic diversity and phylogeographic patterns. Castanopsis (Fagaceae) is one of the dominant tree species in the broadleaved evergreen forests of Japan. In this study we investigate the genetic diversity, genetic structure and leaf epidermal morphology of 63 natural populations of C. sieboldii and C. cuspidata, using 32 Expressed Sequence Tag associated microsatellites. The overall genetic differentiation between populations was low (GST = 0.069 in C. sieboldii and GST = 0.057 in C. cuspidata). Neighbor-joining tree and Bayesian clustering analyses revealed that the populations of C. sieboldii and C. cuspidata were genetically clearly differentiated, a result which is consistent with the morphology of their epidermal cell layers. This suggests that C. sieboldii and C. cuspidata should be treated as independent species, although intermediate morphologies are often observed, especially at sites where the two species coexist. The higher level of genetic diversity observed in the Kyushu region (for both species) and the Ryukyu Islands (for C. sieboldii) is consistent with the available fossil pollen data for Castanopsis-type broadleaved evergreen trees during the Last Glacial Maximum and suggests the existence of refugia for Castanopsis forests in southern Japan. Within the C. sieboldii populations, Bayesian clustering analyses detected three clusters, in the western and eastern parts of the main islands and in the Ryukyu Islands. The west-east genetic differentiation observed for this species in the main islands, a pattern which is also found in several plant and animal species inhabiting Castanopsis forests in Japan, suggests that they have been isolated from each other in the western and eastern populations for an extended period of time, and may imply the existence of eastern refugia. [ABSTRACT FROM AUTHOR]

HepG2 cells were examined for the presence of UDP-D-xylose:β-D-glucoside α-1,3-xylosyltransferase activity by using 2-[(2-pyridyl)amino]ethyl β-D-glucopyranoside (Glcβ-R) as a fluorogenic acceptor. UDP-D-xylose and the acceptor were incubated with the HepG2 microsomal fraction, and the enzymatic reaction mixture was analyzed by HPLC. Structural analysis of the fluorogenic product by α-D-xylosidase digestion, FAB-MS, and Smith degradation revealed that it was Xylα1-3Glcβ-R, thus demonstrating the existence of β-D-glucoside α-1,3-xylosyltransferase. A solubilization study of the enzyme from the microsomal fraction indicated that it differed from UDP-D-xylose:α-D-xyloside α-l,3-xylosyltransferase of the HepG2 microsomal fraction producing Xylα1-3Xylα1-3Glc-R′ [R′, (2-pyridyl)amino-] from UDP-D-xylose and Xylαl-3Glc-R′ [Minamida, S., Aoki, K., Natsuka, S., Omichi, K., Fukase, K., Kusumoto, S. & Hase, S. (1996) J. Biochem. (Tokyo) 120, 1002–1006]. The newly detected enzyme appears to be involved in the biosynthesis of the Xylαl-3Glcβ-Ser structure of glycoproteins such as human blood coagulation factors VII and IX. [ABSTRACT FROM AUTHOR]