Range-wide genetic variation of the widespread cold-temperate spruce Picea jezoensis was studied throughout northeast Asia using maternally inherited mitochondrial DNA and paternally inherited chloroplast DNA markers. This study assessed 33 natural populations including three varieties of the species in Japan, Russia, China, and South Korea. We depicted sharp suture zones in straits around Japan in the geographical distribution pattern of mitochondrial haplotypes (GST=0.901; NST=0.934). In contrast, we detected possible extensive pollen flow without seed flow across the straits around Japan during the past population history in the distribution pattern of chloroplast haplotypes (GST=0.233; NST=0.333). The analysis of isolation by distance of the species implied that by acting as a barrier for the movement of seeds and pollen, the sharp suture zones contributed considerably to the level of genetic differentiation between populations. Constructed networks of mitochondrial haplotypes allowed inference of the phylogeographical history of the species. We deduced that the disjunction with Kamchatka populations reflects range expansion and contraction to the north of the current distribution. Within Japan, we detected phylogeographically different types of P. jezoensis between Hokkaido and Honshu islands; P. jezoensis in Honshu Island may have colonized this region from the Asian continent via the Korean peninsula and the species in Hokkaido Island is likely to have spread from the Asian continent via Sakhalin through land bridges. Japanese endemism of mitochondrial haplotypes in Hokkaido and Honshu islands might have been promoted by separation of these islands from each other and from the Asian continent by the straits during the late Quaternary.

Range-wide genetic variation of Korean pine (Pinus koraiensis) was assessed using maternally inherited mtDNA and paternally inherited cpDNA for 16 natural populations throughout northeast Asia in order to study its phylogeographical history during the Quaternary. The cpDNA variation indicated that there was no difference between populations on the Asian continent and those in the Japanese archipelago. In contrast, the mtDNA variation indicated that there was significant difference between the populations from the two regions, with each region having a different lineage. The continental populations exhibited no diversity in the mtDNA examined despite the species' current extensive range and large populations. Conversely, while the Korean pine is rare in Japan, the Japanese populations exhibited greater levels of mtDNA diversity (H (T) = 0.502). The higher mtDNA diversity and evidence from numerous Korean pine macrofossil remains dated to the Pleistocene and recovered various sites in Japan suggest that the Japanese archipelago once served as a refugium to a much larger Korean pine population with a more extensive range than is the case today. The presence of the single mtDNA haplotype across the Asian continent suggests that the present widespread populations could have expanded from a single refugium population after the last glacial periods.

Aim We used microsatellite markers to determine the range-wide genetic structure of Picea jezoensis and to test the hypothesis that the past population history of this widespread cold-temperate spruce has resulted in a low level of genetic variation and in imprints of inbreeding and bottlenecks in isolated marginal populations.Location The natural range of the three infraspecific taxa of P. jezoensis throughout north-east Asia, including isolated marginal populations. MethodsWe analysed a total of 990 individuals across 33 natural populations using four nuclear microsatellite loci. Population genetic structure was assessed by analysing genetic diversity indices for each population, examining clustering (model-based and distance-based) among populations, evaluating signals of recent bottlenecks, and testing for isolation by distance (IBD). ResultsThe 33 populations were clustered into five groups. The isolated marginal groups of populations (in Kamchatka, Kii in Japan and South Korea) exhibited low levels of allelic richness and gene diversity and a complete or almost complete loss of rare alleles. A recent bottleneck was detected in the populations in Hokkaido across to mid-Sakhalin. The IBD analysis revealed that genetic divergence between populations was higher for populations separated by straits.Main conclusions Picea jezoensis showed a higher level of genetic differentiation among populations (F ST = 0.101) than that observed in the genus Picea in general. This might be attributable to the fact that historically the straits around Japan acted as barriers to the movement of seeds and pollen. The low levels of genetic diversity in the isolated marginal population groups may reflect genetic drift that has occurred after isolation. Evidence of a significant bottleneck between the Hokkaido and mid-Sakhalin populations implies that the cold, dry climate in the late Pleistocene resulted in the decline and contraction of populations, and that there was a subsequent expansion followed by a founder effect when conditions improved. The high polymorphism observed in P. jezoensis nuclear microsatellites revealed cryptic genetic structure that organellar DNA markers failed to identify in a previous study.

Since the 1990s, increasing populations of a blood feeding land leech (Haemadipsa japonica) have become a serious issue in several Japanese prefectures, and it may be caused by the increases in sika deer (Cervus nippon) populations seen over the last quarter of the century. Therefore, this study aimed to reveal the host animal species of H. japonica using iDNA (vertebrate DNA isolated from invertebrates) and to test the hypothesis that the increasingly widespread distribution of sika deer results in increased H. japonica populations through changes to the host–parasite interface. We amplified mitochondrial DNA 16S ribosome RNA fragments from iDNA isolated from the blood clots of H. japonica collected across Japan. We identified 17 host animal species, including four orders of Mammalia (Carnivora, Artiodactyla, Rodentia, and Lagomorpha) and two orders of Amphibia (Caudata and Anura). The sika deer was the dominant host species of H. japonica. Additionally, the host animal species composition of H. japonica differed according to the presence or absence of sika deer. In the sites where sika deer were not found, Anura (frog) species were the most commonly identified hosts of H. japonica. These results suggest that the increases in H. japonica populations might have occurred via a change in host preference to sika deer. This change might be driven by the increases in sika deer populations and subsequent increase in the frequency that H. japonica uses the sika deer as easy prey, as well as by sika deer providing more reproductive energy per blood meal than blood meal from frog species. The present study suggests that a more widespread distribution of sika deer resulted in an increase in H. japonica through a change in the host–parasite interface. Therefore, management that focuses on decreasing sika deer populations would likely be an effective method for the reduction of H. japonica populations.

The genetic structure of Sakhalin spruce (Picea glehnii) was studied across the natural range of the species, including two small isolated populations in south Sakhalin and Hayachine, by using six microsatellite loci and maternally inherited mitochondrial gene sequences. We also analyzed P. jezoensis, a sympatric spruce in the range. Genetic diversity of P. glehnii was higher in central Hokkaido and the lowest in the Hayachine. Bayesian clustering and principal coordinate analysis by using the microsatellites indicated that the Hayachine was clearly distinct from other populations, implying that it had undergone strong genetic drift since the last glacial period. P. glehnii harbored four mitochondrial haplotypes, two of which were shared with P. jezoensis. One of the two was observed without geographical concentration, suggesting its derivation from ancestral polymorphism. Another was observed in south Sakhalin and in P. jezoensis across Sakhalin. The Bayesian clustering--by using four microsatellite loci, including P. jezoensis populations--indicated unambiguous species delimitation, but with possible admixture of P. jezoensis genes into P. glehnii in south Sakhalin, where P. glehnii is abundantly overwhelmed by P. jezoensis; this might explain the occurrence of introgression of the haplotype of P. jezoensis into P. glehnii.