We have identified multiple alleles for a single gene termed W14/15. This gene encodes closely related but not identical proteins W14 and W15 that accumulate in overwinter buds of Gentiana triflora (Takahashi et al. in Breed Sci 56:39-46, 2006; Hikage et al. in Mol Genet Genomics 278:95-104, 2007). In this study, structural analysis of the W14/15 gene was carried out for 21 different gentian lines/cultivars consisting of 5 different species, to survey species- or line/cultivar-specific haplotypes. Within the samples examined, multiple variant forms were found. Those were categorized into seven major types (type I-VII) and ten subtypes based on the presence of three short insertion/deletion sites, three RFLP sites, and several SNP sites. Each line/cultivar had a distinct set of W14/15 gene variants for an allelic pair. Phylogenetic analysis showed that the W14/15 alleles cluster into groups that are characteristic of gentian species, i.e., G. triflora, G. scabra, G. pneumonanthe, G. septemfida and an unknown species other than the former four. In addition, within the same gentian species, different sets of haplotypes were found. Thus, the W14/15 alleles provide useful landmarks to resolve phylogenies of the genus or section Gentiana, as well as to analyze pedigree and breeding history of the cultivars derived from those Gentiana sp.

Extracts from the dried roots of gentian plant, Gentiana triflora, exhibit an antiproliferative activity against cultured and implanted tumor cells. However, the underlying mechanism has been unclear. In the present study, we show that the cell death induced by the extract occurs caspase-independently and depends on metabolic status of mitochondrial respiration. We observed that sensitivity to the extract was considerably lower in HeLa cells, which have a low rate of mitochondrial respiration, in comparison to Y3-Ag1.2.3 cells, which have a higher rate of respiration. Furthermore, sensitivity of HeLa cells to the extract increased significantly when they were forced to switch their energy dependency from glycolysis to mitochondrial respiration. These results indicate that the gentian extract targets on mitochondrial respiration. Consequently, different respiratory activities in mitochondria confer cells to have different susceptibilities to the extract-induced cell death.

The effect of bark from Cryptomeria japonica trees on circadian variations in lymphocyte and neutrophil counts in sheep was studied. Hay supplemented with bark caused decreased leukocyte counts in peripheral blood compared with hay alone. Percentages of lymphocytes were slightly increased, and neutrophils were slightly decreased compared to controls. Plasma NEFA concentrations with bark and hay were also lower than those with hay alone. These results suggest that feed supplemented with bark may increase parasympathetic activity in sheep.