The genus Entada (Leguminosae; Mimosoideae) in the Ryukyu Islands was studied based on number of leaflets, seed morphology and SSCP analysis of chloroplast DNA (rps16 intron). Morphological and molecular data indicated that the Entada in Okinawa Isl. and Yaeyama Isls. was distinctly separated from that in Yaku and Amami-Oshima Islands. We made a deduction that the former having small and convex seeds with angular margin was E. koshunensis, while the latter having large and compressed seeds with rounded margin and symapomorphic insertion (7bp) in the rps16 was E. phaseoloides.
Methyl bromide has been widely used for the fumigation of herbarium specimens, but will be banned by January 2005 under the Montreal Protocol. We recently reported that methyl iodide (CH_3I), as an alternative fumigant, exerted small influence on DNA contained in biological specimens. In order to determine the optimum condition of fumigation against herbarium specimens, seven treatments, combinations of different concentration of CH_3I, temperature and fumigation time in 65% relative humidity, were performed. Fumigation with CH_3I did not produce double-strand breaks of the purified DNA. A main effect of CH_3I on specimen DNA is that the extraction of high-molecular DNAs is obstructed by denatured proteins. Therefore, proteinases added to extract solutions and chloroform extraction facilitate the efficient purification of DNA from fumigated specimens. In all CH_3I treatments, fragments larger than 1,000 bases were amplified using isolated DNAs from fumigated specimens, and they were applicable to molecular phylogenetic analyses. Fumigation of specimens under the condition of 40g/m^3 CH_3I for 24 hours at 25℃ exhibited 100% insecticidal, ovicidal and fungicidal effects and the least influence on specimen DNA. CH_3I exerted little influence on specimen DNA, when fumigation was performed only for insecticidal and ovicidal effects, 20-40g/m^3 CH_3I for 24 hours at 10℃.
Five species and two forms of the genus Lecanorchis were found in Aichi prefecture. L. japonica and L. hokurikuensis (sometimes treated as a vaeiety of the former), and L. kiusiana var. kiusiana and var. suginoana, were morphologically discontinuous from one another in the sympatric habitat (s). They should be separated at the rank of species, respectively. L. kiiensis was considered to be a yellow-flowered form of L. hokurikuensis. A similar yellow-flowered form was also found in L. kiusiana var. suginoana. Consequently, two new names (L. hokurikuensis form, kiiensis and L. suginoana) were proposed, and a new form (L. suginoana form, flava) was described.
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