Tsuneo Funamoto

DNA Barcoding of Panax Species

Ginsengs (Panax, Araliaceae) are among the plants best known for their medicinal properties. Many ginseng species are endangered due to over-exploitation of natural resources - a situation difficult to remedy while there are no reliable, practical methods for species identification. We screened eleven candidate DNA barcoding loci to establish an accurate and effective Panax species identification system, both for commercial and conservation purposes. We used 95 ginseng samples, representing all the species in the genus. We found considerable differences in the performance of the potential barcoding regions. The sequencing of ATPF-ATPH was unsuccessful due to poly-N structures. The RBCL, RPOB, and RPOC1 regions were found to be mostly invariable, with only four to eight variable sites. Using MATK, PSBK-I, PSBM-TRND, RPS16 and NAD1, we could identify four to six out of eight considerably divergent species but only one to five out of nineteen clusters within the P. bipinnatifidus species group. PSBA-TRNH and ITS were the most variable loci, working very well both in species and cluster identifications. We demonstrated that the combination of PSBA-TRNH and ITS is sufficient for identifying all the species and clusters in the genus.

Molecular Phylogeny and Biogeographic Diversification of Linnaeoideae (Caprifoliaceae s. l.) Disjunctly Distributed in Eurasia, North America and Mexico

Linnaeoideae is a small subfamily of erect or creeping shrubs to small trees in Caprifoliaceae that exhibits a wide disjunct distribution in Eurasia, North America and Mexico. Most taxa of the subfamily occur in eastern Asia and Mexico but the monospecific genus Linnaea has a circumboreal to north temperate distribution. In this study, we conducted phylogenetic and biogeographic analyses for Linnaeoideae and its close relatives based on sequences of the nuclear ribosomal ITS and nine plastid (rbcL, trnS-G, matK, trnL-F, ndhA, trnD-psbM, petB-D, trnL-rpl32 and trnH-psbA) markers. Our results support that Linnaeoideae is monophyletic, consisting of four eastern Asian lineages (Abelia, Diabelia, Dipelta and Kolkwitzia), the Mexican Vesalea, and Linnaea. The Mexican Vesalea was formerly placed in Abelia, but it did not form a clade with the eastern Asian Abelia; instead Vesalea and Linnaea are sisters. The divergence time between the eastern Asian lineages and the Mexican Vesalea plus the Linnaea clade was dated to be 50.86 Ma, with a 95% highest posterior density of 42.8 Ma (middle Eocene) to 60.19 Ma (early Paleocene) using the Bayesian relaxed clock estimation. Reconstructed ancestral areas indicated that the common ancestor of Linnaea plus Vesalea may have been widespread in eastern Asia and Mexico or originated in eastern Asia during the Eocene and likely migrated across continents in the Northern Hemisphere via the North Atlantic Land Bridges or the Bering Land Bridge. The Qinling Mountains of eastern Asia are the modern-day center of diversity of Kolkwitzia-Dipelta-Diabelia clade. The Diabeliaclade became highly diversified in Japan and eastern China. Populations of Diabelia serrata in Japan and eastern China were found to be genetically identical in this study, suggesting a recent disjunction across the East China Sea, following the last glacial event.

Molecular phylogenetics of Chelonopsis ( Lamiaceae: Gomphostemmateae) as inferred from nuclear and plastid DNA and morphology

The recently established tribe Gomphostemmateae (Lamiaceae) consists of Gomphostemma, Bostrychanthera and Chelonopsis, but relationships within and between these genera have not been well studied. Only six of the approximately 46 species of Gomphostemmateae have been included in any previous phylogenetic analysis. In the present study, we used DNA sequences of two nuclear regions (ITS, ETS) and five plastid loci (psbA-trnH, rps 16, trnL intron, trnL-trnF spacer, trnStrnG) to reconstruct the phylogeny of the East Asian genus Chelonopsis for the first time. The molecular results indicate that the tribe Gomphostemmateae is monophyletic; Bostrychanthera is embedded in Chelonopsis; and Chelonopsis comprises two clades, one encompassing the taxa of Chelonopsis subg. Chelonopsis and the genus Bostrychanthera and the other consisting of Chelonopsis subg. Aequidens. This split is supported by several morphological characters. These results, which are further strengthened by morphological and cytological data, indicate that Bostrychanthera should be transferred to Chelonopsis. In addition, our results show that within C. subg. Aequidens, sect. Aequidens and sect. Microphyllum are monophyletic. Furthermore, two major clades are concordant with the Sino-Japanese and Sino-Himalayan distribution patterns..

Pollen morphology of the East Asiatic genus Chelonopsis (Lamioideae: Lamiaceae) and allied genera, with reference to taxonomic implications and potential pollination ecology

Pollen grains of 15 taxa of the genus Chelonopsis (14 spp. and 1 variety) from China and Japan and 6 species of the closely related genera Bostrychanthera (1 species) and Gomphostemma (5 species) were examined by light and scanning electron microscopy. Of these, the pollen morphology of 18 taxa was studied for the first time. Pollen grains were found to be tricolpate with polar lengths of 20.8–30.0 μm and equatorial widths of 17.5–27.3 μm. The basic shape of the pollen in most taxa is subprolate or prolate-spheroidal, but spheroidal, subprolate-spheroidal, oblate-spheroidal, and prolate-subprolate grains are also found in some species. The surface is generally reticulate or suprareticulate in Chelonopsis and granulate in Bostrychanthera. In comparison with those of Chelonopsis and Bostrychanthera, the pollen grain surfaces of Gomphostemma are more diverse. In Chelonopsis, pollen is taxonomically useful at the sectional level, and some grains provide enough characters for species delimitation. The potential pollination ecology of cellular hairs on the anthers of Chelonopsis and Bostrychanthera is also briefly discussed.

Karyotypic study of eighteen taxa of Salvia (Lamiaceae) from China

Abstract Karyotypes of 16 species and two varieties of Chinese Salvia were determined in this study. Of the 18 Salvia taxa, chromosome numbers of eight taxa and karyotypic analyses of 13 taxa were reported for the first time. The basic chromosome number of all taxa in this study was x = 8. With the exception of tetraploids S. campanulata var. codonantha and S. przewalskii, the taxa were diploid. One B chromosome was found in part of the mitosis metaphase cells of S. aerea. The present results were compared with previous cytological studies. The population of S. castanea having x = 11 from India may be misidentified. Consequently, all Salvia taxa from China may have the same basic chromosome number of x = 8. The population of S. przewalskii reported as tetraploid from Russia and populations of S. evansiana reported as tetraploid from Yunnan and Sichuan of China were very likely to be misidentified. Therefore, S. przewalskii may be tetraploid without diploid representatives, and S. evansiana diploid without any tetraploids. Salvia campanulata is both diploid and tetraploid. The B chromosome is rather unsteady among populations of the same species, even within the same tissues.

Chromosome numbers of 24 taxa of Lamiaceae from Southwest China

ABSTRACT Lamiaceae is the sixth largest angiosperm family and comprises over 7000 species in 236 genera distributed worldwide. China is one of the diversity centers of Lamiaceae; however, limited chromosome counts are available for Chinese Lamiaceae species. In the present study, chromosome numbers of 32 populations representing 21 species and three varieties of nine genera of Lamiaceae from Southwest China are studied, among which chromosome counts of 17 taxa are reported for the first time. Our results show that most of the species studied are diploids, and are consistent with the previously proposed base numbers. Three species, Isodon ternifolius (2n = 48), Nepeta dentata (2n = 32), and one population of Dracocephalum heterophyllum (2n = 24), are shown to be tetraploids. Chromosome numbers for 56 taxa of 11 genera of Lamiaceae from the Hengduan Mountains Region are obtained from published literature and present study, and the frequency of polyploidy is only 12.5%. Our study further support that polyploidy may have played a minor role in the evolutionary diversification of Lamiaceae in the Hengduan Mountains.