Dadashipour M.,Toyama Prefectural University |
Yamazaki M.,Toyama Prefectural University |
Momonoi K.,Toyama Agricultural Research Center |
Tamura K.,Toyama Prefectural University |
And 5 more authors.
Journal of Biotechnology | Year: 2011
A novel S-hydroxynitrile lyase (HNL) was purified from leaves of a plant, Baliospermum montanum, by ammonium sulfate fractionation and column chromatographies. Full-length cDNA and genomic DNA were cloned and sequenced. The latter contained two introns and one ORF encoding a 263-residue protein (subunit: 29.5kDa). The hnl gene was expressed in Escherichia coli and the enzyme was characterized including detailed kinetic studies of 20 substrates for (S)-cyanohydrin synthesis. The enzyme exhibited the highest specific activity (178U/mg), k cat (98/s) and k cat/K m ratio for piperonal. k cat/K m ratio for aromatic aldehydes was much larger than those of aliphatic aldehydes and ketones. It was strongly inhibited by AgNO 3, PMSF, phenol and methyl ethyl ketone, showed an optimum at pH 5, while having activity at range of 4-6.5. It exhibited stability at wide pH range 2.4-11, the highest activity at 20°C, being active at 0-65°C. The enzyme showed variations in residues involved in substrate pocket and substrate entrance channel compared to other S-selective HNLs, based on a model was built. C-terminal short truncations provided more enzyme production. Gel filtration revealed a 60-65kDa molecular mass for this non-FAD enzyme and its C-terminally truncated forms using three buffer compositions, indicating dimeric structures. © 2011 Elsevier B.V.
Orihara T.,Tottori University |
Sawada F.,2 11 12 Tsujido taiheidai |
Ikeda S.,Forest Development Technological Institute |
Yamato M.,General Environmental Technos Co. |
And 5 more authors.
Mycologia | Year: 2010
During taxonomic revision of genus Octaviania in Japan we examined herbarium and fresh specimens of O. columellifera and O. asterosperma sensu S. Yoshimi & Y. Doi with morphological and molecular techniques. These two species were identical in both macro- and micromorphological characters and were clearly different from the generally known O. asterosperma. The identity of the two species and their distinctness from O. asterosperma was further supported by both nuclear large subunit and ITS rDNA phylogeny. The molecular analyses also revealed that O. columellifera shares its lineage with the boletoid mushroom-forming Xerocomus chrysenteron complex and that it does not form a monophyletic clade with other Octaviania species. Our morphological reevaluation, including transmission electron microscopic observation of basidiospores, clarified the taxonomic boundary between O. columellifera and other Japanese Octaviania species. Accordingly we propose a new genus, Heliogaster, for O. columellifera with designation of the lectotype. We discuss phylogenetic relationships with Octaviania sensu stricto species and the closely related boletoid (pileate-stipitate) fungi, generic characters of Helio-gaster and intraspecific phylogeny. © 2010 by The Mycological Society of America.
Godo T.,Botanic Gardens of Toyama |
Komori M.,Botanic Gardens of Toyama |
Nakaoki E.,Toyama Prefectural University |
Yukawa T.,Tsukuba Botanical Garden |
Miyoshi K.,Akita Prefectural University
In Vitro Cellular and Developmental Biology - Plant | Year: 2010
The effects of culture conditions on the asymbiotic germination of mature seeds of Calanthe tricarinata Lindl., an endangered terrestrial cool-climate orchid, were examined. Specifically, conditions such as illumination, temperature, and the addition of plant growth regulators to the medium were studied. Mature seeds were harvested from plants that had been collected in Toyama Prefecture, Japan, and maintained at the Botanic Gardens of Toyama. Solidified "New Dogashima" medium was used as the basal medium, and it was supplemented with 6-benzyladenopurine (BA) or α-naphthalene acetic acid (NAA). White light at 40 μmol m-2 s-1, with a 16-h photoperiod, inhibited the germination of seeds by 53-80%, as compared to dark controls in genotypes examined. The optimal temperature for the germination of seeds in darkness was 20°C and the germination frequency reached 60%, whereas it was only 28% at 25°C. While both NAA and BA stimulated germination, BA was more effective than NAA. After storage for 18 mo at 5°C, seeds incubated on medium that contained 0. 2 mg l-1 BA germinated at a frequency of 36%, which was twice that of seeds grown without any plant growth regulators. The frequency of subsequent germination decreased during storage of seeds at 5°C for approximately 2 yr, dropping from 61% to 13%. The protocorms obtained in this study were developed to plantlets readily after transferring to fresh 1/2 MS medium without any plant growth regulators. They were successfully acclimatized in green house after two to three subcultures in vitro. The significant role of a reproducible protocol for the germination of mature seeds is discussed in terms of the ex situ conservation of endangered orchid species. © 2010 The Society for In Vitro Biology.
Uehara A.,Tokyo University of Agriculture and Technology |
Nakata M.,Botanic Gardens of Toyama |
Kitajima J.,Showa Pharmaceutical University |
Iwashina T.,Tokyo University of Agriculture and Technology
Biochemical Systematics and Ecology | Year: 2012
Flavonoids in the leaves of 17 species and 4 varieties of Japanese Chrysanthemum were surveyed. Nine flavonoid glycosides were isolated and identified as quercetin 3- O-rutinoside, eriodictyol 7- O-glucuronide, luteolin 7- O-rutinoside, 7- O-glucoside and 7- O-glucuronide, apigenin 7- O-glucoside and 7- O-glucuronide, chrysoeriol 7- O-glucuronide and acacetin 7- O-rutinoside. The taxa were divided into three chemotypes by their flavonoid characters, i.e. I) occurrence of flavone, flavonol and flavanone, II) occurrence of flavone and flavanone and III) occurrence of flavone alone. The chemotypes almost agreed with the system based on morphological, cytological and geographical characters, i.e. Group I) the taxa growing on seashore and having tubular flowers alone, Group III) the taxa of the Asian Continent origins and having both tubular and ligulate flowers. Others were included in Group II which was subdivided into four groups by flavonoid characters. Twelve external flavonoids were isolated and identified as acacetin 7- O-rutinoside, luteolin, nepetin, 5,7,3',4'-tetrahydroxy-6,5'-dimethoxyflavone, apigenin, hispidulin, 5,7,4'-trihydroxy-6,3',5'-trimethoxyflavone, jaceosidin, sudachitin, eupatilin, hymenoxin and pectolinarigenin. The distribution patterns of almost all external flavonoids were apparently irregular among the taxa. © 2011 Elsevier Ltd.
Nakano M.,Niigata University |
Kuwayama S.,Niigata University |
Oka E.,Niigata University |
Asano M.,Niigata University |
And 2 more authors.
Journal of the Japanese Society for Horticultural Science | Year: 2013
Lychnis belongs to Caryophyllaceae and contains a number of horticulturally attractive species. In order to widen their variations in horticultural traits, interspecific cross-pollination and subsequent immature seed culture were carried out using 8 Lychnis species, L. chalcedonica, L. coronata, L. fulgens, L. gracillima, L. kiusiana, L. miqueliana, L. sieboldii, and L. wilfordii. Enlarged fruits containing immature seeds were obtained in the 26 cross-combinations 4 weeks after pollination. Immature seeds were isolated from the fruits and cultured on halfstrength Murashige and Skoog medium without plant growth regulators, on which germination occurred in 11 cross-combinations and seedlings were produced in 8 cross-combinations. However, green seedlings were obtained only in L. fulgens × L. sieboldii, and seedlings obtained from the other 7 cross-combinations, L. coronata × L. gracillima, L. gracillima × L. coronata, L. kiusiana × L. fulgens, L. kiusiana × L. sieboldii, L. kiusiana × L. wilfordii, L. wilfordii × L. kiusiana, and L. wilfordii × L. sieboldii, were albino and died during acclimatization. The hybridity of all 55 green seedlings obtained from L. fulgens × L. sieboldii was confirmed by random amplified polymorphic DNA analysis. Although some morphological variations were observed among hybrids, hybrid plants generally showed intermediate morphologies between the parents. Hybrids showed high pollen fertility and their selfpollination yielded viable seeds. The present study shows the possibility of interspecific cross-breeding in Lychnis. Further studies are necessary to improve the culture conditions of immature seeds and to examine interspecific cross-compatibility using a wide range of Lychnis species. © JSHS 2013.
Kanemoto T.,Botanic Gardens of Toyama
Journal of Japanese Botany | Year: 2013
Morphological features of triploid and tetraploid cytotypes of Pellionia scabra were examined. In reproductive characters, the triploid cytotype is distinguishable from the tetraploid cytotype by abundant, smaller achenes and several dorsal setae on the horn-like appendages of the female flower perianth. There are no differences between the two cytotypes in seven vegetative characters examined. Morphological features of the triploid cytotype agree with diagnostic characteristics of P. scabra Benth. van pedimculata Yamam., therefore, this variety is considered to be the triploid cytotype of P scabra.
Uchimura E.,Botanic Gardens of Toyama
Acta Horticulturae | Year: 2013
From my point of view, bamboo is different from woody plants and herbaceous plants, and does not belong to any of these groups because of its intermediary characteristics, which are discussed in this lecture. The geographical distribution of bamboo in the world and its biological and genetic resources are presented. Some factors influencing bamboo growth, as temperature, rainfall, soil, topography, are also discussed. Bamboo is a multipurpose crop and recently its importance as biomass production is improving.
Godo T.,Botanic Gardens of Toyama |
Lu Y.,Botanic Gardens of Toyama |
Mii M.,Botanic Gardens of Toyama
Methods in molecular biology (Clifton, N.J.) | Year: 2010
Numerous shoots were directly regenerated from the leaf explants of Lysionotus pauciflorus on the MS medium containing 0.5-2 microM NAA with or without 1 microM BA. The calli were induced from the leaves on MS medium supplemented with 2 microM 2, 4-D. The calli proliferated about four times in fresh weight in the liquid medium of the same composition as the callus induction medium after 4 weeks of culture on a rotary shaker at 100 rpm. Shoots were induced from these calli on the regeneration medium amended with 32 microM BA or 0.5 microM zeatin. Regenerated shoots rooted easily on (1/2) MS medium without any plant growth regulators. Most of the regenerants from callus were diploid, whereas eight of 66 acclimatized plantlets were tetraploid determined by flow cytometric analysis. Furthermore, flow cytometric analysis of calli also revealed tetraploidy.
Godo T.,Botanic Gardens of Toyama |
Fujiwara K.,University of Tokyo |
Guan K.,CAS Kunming Institute of Botany |
Miyoshi K.,Akita Prefectural University
Plant Biotechnology | Year: 2011
The effects of different light wavelengths on in vitro asymbiotic germination of mature seeds and seedling growth of Bletilla ochracea Schltr. were examined using five wavelengths from light emitting diodes with peak wavelengths of 470 nm (blue), 525 nm (green), 590 nm (orange), 625 nm (red), and from white light emitting diodes at 40μ μmolm -2 s -1. Gellan gum solidified New Dogashima medium without plant growth regulators was used as germination medium. Cultures were maintained at 25°C under 24 h continuous lighting or darkness. The frequencies of seed germination three weeks after sowing reached more than 60% in all light conditions examined including continuous darkness. The highest frequencies of seed germination of 74% were achieved using lighting both with green and orange light emitting diodes, though strong inhibition of seed germination by the specific wavelength was not recognized. Seedling growth of this species was greatly inhibited by darkness. The most effective wavelength of light for rhizoid formation was revealed to be in the range of 590 nm (orange light) and 625 nm (red light), and almost no rhizoid was formed in the darkness. After 3 months of culture, leave width was expanded under white and blue light emitting diodes and was narrower under green, orange and red light. Seedlings grown under white and blue light emitting diodes resulted in thicker pseudobulbs. © 2011 The Japanese Society for Plant Cell and Molecular Biology.
Nakata M.,Botanic Gardens of Toyama
Journal of Japanese Botany | Year: 2014
Although it has long been used conventionally in much of the literature, the representation of chromosome number as "n =," which is based on the observation of bivalent chromosomes at diakinesis or at meiotic metaphase I in the pollen mother cell, is incorrect. Since the meiocyte itself is diploid, the chromosome number should be described as the "2n =" number, for example, 2n = 9 II = 18 when nine bivalents are observed in the meiotic course. The haploid chromosome number "n =" should be given by the direct observation of chromosomes in the gametophyte, for example, the pollen grain or pollen tube of seed plants, the prothallium of ferns, and the vegetative plant of bryophytes.