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The genus Asarum (Aristolochiaceae) encompasses approximately 120 species from five sections. Taxonomic controversies concerning the genus Asarum and/or its intrageneric classification remain unresolved. In particular, sect. Heterotropa accounts for a large percentage of the genus (80 of 120 species) and is well diverged in the Sino-Japanese Forest subkingdom. Reconstruction of Heterotropa phylogeny and estimation of its divergence times would provide significant insight into the process of species diversity in the Sino-Japanese floristic region. This study encompassed 106 operational taxonomic units (OTUs), and phylogenetic analyses were conducted based on internal transcribed spacer (ITS) and matK sequences. Although the matK sequences provided informative results solely for section Geotaenium, phylogenetic trees based on ITS regions yielded a clear result for several sections. Three sections, Asarum, Geotaenium and Asiasarum, were supported as robust monophyletic groups, whereas Heterotropa had low support. Sect. Hexastylis was revealed to be polyphyletic, suggesting taxonomic reconstruction would be needed. Sect. Heterotropa comprises two clades, which correspond to species distribution ranges: mainland China and the island arc from Taiwan to mainland Japan via the Ryukyu Islands. It is notable that the common ancestry of the latter clade in the eastern Asian islands was highly supported, suggesting that the present species diversity of Heterotropa was initially caused by allopatric range fragmentation in East Asia. [ABSTRACT FROM AUTHOR]

A set of 11 polymorphic microsatellite markers has been developed and characterized for the critically endangered species Pieris amamioshimensis. Fifty-nine individuals of an ex-situ population were used to identify these markers. The total number of alleles for each locus ranged from 3 to 9, with an average of 5.4. The expected heterozygosities ( HS) and observed heterozygosities ( HO) ranged from 0.47 to 0.77 and 0.22 to 0.88, respectively. In total, four loci exhibited significant deviations from Hardy-Weinberg equilibrium: two loci showed significant heterozygosity excess and the other two loci showed significant heterozygosity deficit. The polymorphism information content (0.43 ≤ PIC ≤ 0.73), the probability of exclusions (PE1 = 0.9565, PE2 = 0.9969 and PE3 = 0.9999) and probabilities for identity (PI = 3.78 × 10−9 and PI-Sib = 2.35 × 10−4) suggest that these markers are useful for estimating not only genetic diversity but also parentage, for the ex-situ conservation management of populations. [ABSTRACT FROM AUTHOR]

Primula tosaensis (Primulaceae) is an endangered primrose endemic to Japan. In this study, 24 novel microsatellite markers were developed using Illumina MiSeq sequencing to facilitate conservation of this endangered species. The genetic diversity and polymorphisms of these novel markers were measured in 32 individuals from a wild P. tosaensis population. The number of alleles and expected heterozygosities ranged from 2 to 5 (mean = 2.8) and from 0.119 to 0.724 (mean = 0.395), respectively. All loci were in Hardy-Weinberg equilibrium. The markers developed in this study will provide a powerful and practical tool for investigating the population structure and genetic diversity of P. tosaensis. [ABSTRACT FROM AUTHOR]

The genus Tanakaea is a plant genus that consists of one or two evergreen herbaceous species in Japan and China. As rithophytic plant species occur on shaded rocks, the populations are usually isolated and sporadically found in disjunct areas. To evaluate the genetic structure of the species at multiple spatial scales, 10 nuclear and mitochondrial microsatellite markers were developed. The novel markers showed high genetic variations (two to 15 alleles and He from 0.400 to 0.894). Clonal samples were identified with the probability of identity of 9.0E-8. When evaluated with 11 populations in Japan, significant genetic differentiation between regional population groups was detected ( FST = 0.313 between Shikoku and Honshu islands), suggesting they have long been isolated from each other. Overall, these markers will be useful for population genetic research to investigate clonal structure and genetic diversity and levels of genetic differentiation between the geographically isolated populations. [ABSTRACT FROM AUTHOR]

EST-SSR markers were developed for A sarum sakawanum var. stellatum to evaluate the genetic diversity and genetic structure of series S akawanum and assess their utility in other series in the genus Asarum. A total of 1864 primer pairs, including 1864 tri-nucleotide SSR motifs, were designed based on transcriptome data. Of the 96 primers tested, 17 loci were successfully amplified and polymorphic in series S akawanum taxa. Furthermore, all these loci were successfully amplified in A. takaoi, whereas only 10 loci were amplified in the more phylogenetically distant A . pellucidum. For populations of A . sakawanum var. stellatum, the values of observed and expected heterozygosity ranged from 0.261 to 0.957 and 0.317 to 0.881, respectively. These newly developed EST-SSR markers will be used in population genetic studies of series S akawanum to infer the evolutionary history and genetic diversity of these series, many of which are endangered in Japan. [ABSTRACT FROM AUTHOR]