星野 洋一郎 (ホシノ ヨウイチロウ)

北方生物圏フィールド科学センター 耕地圏ステーション 生物生産研究農場教授
Last Updated :2025/02/13

■研究者基本情報

学位

  • 博士(学術), 千葉大学

Researchmap個人ページ

研究キーワード

  • 植物育種学
  • 園芸学
  • Plant Breeding
  • Horticulture

研究分野

  • 環境・農学, 遺伝育種科学
  • 環境・農学, 園芸科学

担当教育組織

■経歴

学内役職歴

  • 北方生物圏フィールド科学センター 副センター長, 2022年4月1日 - 2024年3月31日

■研究活動情報

論文

  • Effects of the sucrose concentrations and incubation periods on in vitro pollen germination and pollen tube growth in three rice cultivars
    Neema Yona Yohana, Arisa Nakano, Yoichiro Hoshino
    Plant Biotechnology, 2024年12月24日, [査読有り], [最終著者, 責任著者]
    英語, 研究論文(学術雑誌)
  • Elucidating the impact of ploidy level on biochemical content accumulation in haskap (Lonicera caerulea L. subsp. edulis (Turcz. ex Herder) Hultén) fruits: A comprehensive approach for fruit assessment
    Jixiao Li, Yoichiro Hoshino
    Scientia Horticulturae, 2024年03月, [査読有り], [最終著者, 責任著者]
    研究論文(学術雑誌)
  • Distinctive development of embryo and endosperm caused by male gametes irradiated with carbon-ion beam
    Tomonari Hirano, Muneaki Murata, Yurie Watarikawa, Yoichiro Hoshino, Tomoko Abe, Hisato Kunitake
    Plant Reproduction, 2024年02月09日, [査読有り]
    英語, 研究論文(学術雑誌), Abstract
    Key message
    In Cyrtanthus mackenii, development of embryo and endosperm were differentially affected by fertilization of male gametes with DNA damage and mutations.

    Abstract
    Pollen irradiation with ionizing radiations has been applied in plant breeding and genetic research, and haploid plant induction has mainly been performed by male inactivation with high-dose irradiation. However, the fertilization process of irradiated male gametes and the early development of embryo and endosperm have not received much attention. Heavy-ion beams, a type of radiation, have been widely applied as effective mutagens for plants and show a high mutation rate even at low-dose irradiation. In this study, we analyzed the effects of male gametes of Cyrtanthus mackenii irradiated with a carbon-ion beam at low doses on fertilization. In immature seeds derived from the pollination of irradiated pollen grains, two types of embryo sacs were observed: embryo sac with a normally developed embryo and endosperm and embryo sac with an egg cell or an undivided zygote and an endosperm. Abnormalities in chromosome segregation, such as chromosomal bridges, were observed only in the endosperm nuclei, irrespective of the presence or absence of embryogenesis. Therefore, in Cyrtanthus, embryogenesis is strongly affected by DNA damage or mutations in male gametes. Moreover, various DNA contents were detected in the embryo and endosperm nuclei, and endoreduplication may have occurred in the endosperm nuclei. As carbon-ion irradiation causes chromosomal rearrangements even at low doses, pollen irradiation can be an interesting tool for studying double fertilization and mutation heritability.
  • Agronomic performance in tetraploid Lilium leichtlinii: larger flowers and earlier flowering
    Masumi Yamagishi, Yutaka Jitsuyama, Yoichiro Hoshino
    Euphytica, 219, 12, Springer Science and Business Media LLC, 2023年11月16日, [査読有り]
    英語, 研究論文(学術雑誌)
  • Automated estimation of stomatal number and aperture in haskap (Lonicera caerulea L.)
    Xiangji Meng, Arisa Nakano, Yoichiro Hoshino
    Planta, 2023年10月, [査読有り], [最終著者, 責任著者]
    研究論文(学術雑誌)
  • Deep learning-based high-throughput detection of in vitro germination to assess pollen viability from microscopic images
    Mengwei Zhang, Jianxiang Zhao, Yoichiro Hoshino
    Journal of Experimental Botany, 2023年08月16日, [査読有り], [最終著者, 責任著者]
    研究論文(学術雑誌)
  • Phenotypic analysis of polyploid and aneuploid haskap (Lonicera caerulea L. subsp. edulis (Turcz. ex Herder) Hultén) plants and their progeny production
    Jixiao Li, Yumeno Suzuki, Yoichiro Hoshino
    Euphytica, 2023年05月, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Novel production of β-cryptoxanthin in haskap (Lonicera caerulea subsp. edulis) hybrids: Improvement of carotenoid biosynthesis by interspecific hybridization
    Ryohei Fujita, Shigeki Jin, Kotaro Matoba, Yoichiro Hoshino
    Scientia Horticulturae, 2023年01月, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Production of tetraploid and octoploid Haemanthus albiflos plants using immature embryo-derived embryogenic calli treated with colchicine
    Arisa Nakano, Yoichiro Hoshino
    Plant Cell, Tissue and Organ Culture (PCTOC), 2022年03月13日, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Evaluation of pollen tube growth ability in Petunia species having different style lengths
    Miyako Kato, Hitoshi Watanabe, Yoichiro Hoshino
    Plant Biotechnology, Japanese Society for Plant Cell and Molecular Biology, 2022年, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌)
  • Simultaneous production of triploid and hexaploid plants by endosperm culture with colchicine treatment in diploid Haemanthus albiflos
    Arisa Nakano, Masahiro Mii, Yoichiro Hoshino
    Plant Cell, Tissue and Organ Culture (PCTOC), 2020年11月27日, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Comparison of anthocyanin distribution in berries of Haskap (Lonicera caerulea subsp. edulis (Turcz. ex. Herder) Hultén), Miyama-uguisukagura (Lonicera gracilipes Miq.), and their interspecific hybrid using imaging mass spectrometry
    Ryohei Fujita, Takahiro Hayasaka, Shigeki Jin, Shu-Ping Hui, Yoichiro Hoshino
    Plant Science, 300, 110633, 110633, Elsevier {BV}, 2020年11月, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Phylogeny and trait variation of Japanese Rubus subgenus Ideaobatus
    Ayana Okada, Satoshi Kikuchi, Yoichiro Hoshino, Hisato Kunitake, Makiko Mimura
    Scientia Horticulturae, 264, 109150, 109150, Elsevier {BV}, 2020年04月, [査読有り]
    研究論文(学術雑誌)
  • Evaluation of Fruit Anthocyanin Composition by LC/MS in Interspecific Hybrids Between Haskap (Lonicera caerulea subsp. edulis (Turcz. ex. Herder) Hultén) and Miyama-uguisukagura (Lonicera gracilipes Miq.)
    Ryohei Fujita, Shigeki Jin, Takahiro Hayasaka, Kotaro Matoba, Yoichiro Hoshino
    The Horticulture Journal, 2020年, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌)
  • Estimation of chromosome number among the progeny of a self-pollinated population of triploid Senno (Lychnis senno Siebold et Zucc.) by flow cytometry
    Yoichiro Hoshino, Masashi Nakata, Toshinari Godo
    Scientia Horticulturae, 256, 108542, 108542, Elsevier BV, 2019年10月, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Regulation of dedifferentiation and differentiation in different explants of Papaver rhoeas L. by one-step culture
    Zahra Aghaali, Yoichiro Hoshino, Sajad Rashidi Monfared, Ahmad Moieni
    Scientia Horticulturae, 246, 366, Elsevier {BV}, 2019年02月, [査読有り], [責任著者]
  • Resistance of Sclerotinia homoeocarpa field isolates to succinate dehydrogenase inhibitor fungicides
    James T. Popko, Hyunkyu Sang, Jaemin Lee, Toshihiko Yamada, Yoichiro Hoshino, Geunhwa Jung
    Plant Disease, 102, 12, 2625, 2631, 2018年12月, [査読有り]
    英語, 研究論文(学術雑誌)
  • Nucleic adaptability of heterokaryons to fungicides in a multinucleate fungus, Sclerotinia homoeocarpa
    Dylan Kessler, Hyunkyu Sang, Amanda Bousquet, Jonathan P. Hulvey, Dawlyn Garcia, Siyeon Rhee, Yoichiro Hoshino, Toshihiko Yamada, Geunhwa Jung
    Fungal Genetics and Biology, 115, 64, 77, Academic Press Inc., 2018年06月01日, [査読有り]
    英語, 研究論文(学術雑誌), Sclerotinia homoeocarpa is the causal organism of dollar spot in turfgrasses and is a multinucleate fungus with a history of resistance to multiple fungicide classes. Heterokaryosis gives rise to the coexistence of genetically distinct nuclei within a cell, which contributes to genotypic and phenotypic plasticity in multinucleate fungi. We demonstrate that field isolates, resistant to either a demethylation inhibitor or methyl benzimidazole carbamate fungicide, can form heterokaryons with resistance to each fungicide and adaptability to serial combinations of different fungicide concentrations. Field isolates and putative heterokaryons were assayed on fungicide-amended media for in vitro sensitivity. Shifts in fungicide sensitivity and microsatellite genotypes indicated that heterokaryons could adapt to changes in fungicide pressure. Presence of both nuclei in heterokaryons was confirmed by detection of a single nucleotide polymorphism in the β-tubulin gene, the presence of microsatellite alleles of both field isolates, and the live-cell imaging of two different fluorescently tagged nuclei using laser scanning confocal microscopy. Nucleic adaptability of heterokaryons to fungicides was strongly supported by the visualization of changes in fluorescently labeled nuclei to fungicide pressure. Results from this study suggest that heterokaryosis is a mechanism by which the pathogen adapts to multiple fungicide pressures in the field.
  • Respiratory burst oxidase-D expression and biochemical responses in Festuca arundinacea under drought stress
    Iman Rohollahi, Nayer A. Khoshkholghsima, Hironori Nagano, Yoichiro Hoshino, Toshihiko Yamada
    Crop Science, 58, 1, 435, 442, Crop Science Society of America, 2018年01月01日, [査読有り]
    英語, 研究論文(学術雑誌), Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases catalyze the production of superoxide, a type of reactive oxygen species (ROS). In plants, the NADPH oxidase homologs have been identified as respiratory burst oxidase homologs (Rboh). They are involved in ROS production in response to drought stress. Three entries of tall fescue (Festuca arundinacea Schreb.) were used for analyses in the present study: tolerant (‘Isfahan’) and sensitive (‘Quchan’) accessions to drought during the germination stage, selected from 14 wild populations in Iran, as well as ‘Barvado’ as a control. Partial sequence of the Festuca respiratory burst oxidase-D (FrbohD) gene was isolated from Barvado. We compared expression levels of the FrbohD gene as well as hydrogen peroxide (H2O2), catalase activity and some biochemical responses among the three entries. Gene expression was evaluated for leaf and shoot samples subjected to 3, 6, and 9 d without water. The transcript level of FrbohD, H2O2 content, and catalase activity increased in Quchan under drought stress. It appears that lower levels of FrbohD gene transcription and H2O2 concentration in F. arundinacea leaves contributed to drought stress tolerance in Isfahan. Total protein and total soluble carbohydrate content also increased significantly in Isfahan when it was subjected to drought stress. Isfahan exhibited drought resistance through various strategies, which could serve as selection criteria for improving drought resistance in turfgrass breeding programs.
  • Evaluation of female gamete fertility through histological observation by the clearing procedure in Lilium cultivars
    Yoichiro Hoshino, Noriko Kanematsu, Masahiro Mii
    Breeding Science, 68, 3, 360, 366, Japanese Society of Breeding, 2018年, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Interspecific Hybridization Using Miyama Matatabi (Actinidia kolomikta), a Japanese Indigenous Wild Kiwifruit Relative
    Issei Asakura, Yoichiro Hoshino
    The Horticulture Journal, 87, 4, 481, 489, Japanese Society for Horticultural Science, 2018年, [査読有り], [責任著者]
    研究論文(学術雑誌)
  • Evaluation of plant regeneration ability of different explants and establishment of an efficient regeneration system using immature seeds in Actinidia kolomikta, a cold-hardy kiwifruit relative
    Issei Asakura, Yoichiro Hoshino
    SCIENTIA HORTICULTURAE, 220, 275, 282, ELSEVIER SCIENCE BV, 2017年06月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Efficient protocols for plant regeneration systems are useful for genetically improving Actinidia species using biotechnological approaches such as somatic hybridization, gene transfer, and in vitro chromosome doubling. In this study, we used diploid Actinidia kolomikta (Maxim. &Rupr.) Maxim., a cold-hardy kiwifruit relative, to establish a plant regeneration system. Immature seeds, leaves, petioles, stems, and roots were examined to evaluate callus induction and plant regeneration ability. As a result, all explants on Murashige and Skoog medium with 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin exhibited callus formation at different frequencies varying from 42.4% to 100%. Among them, root-derived and immature seed -derived calli exhibited high shoot regeneration ability at 41.7% and 9.7%, respectively. Immature seed -derived callus produced a higher number of shoots and showed vigorous growth. Therefore, we used immature seed -derived callus to characterize the regeneration process and evaluated the regenerated plants. Cultured immature seeds exhibited two types of calli: compact (white, compact, and unable to regenerate) and soft (white, soft, composed of easily separable mixed granular and nodular structures). When the soft callus was transferred to MS medium with zeatin, the nodular structures developed adventitious shoots, and these shoots developed many adventitious roots on half-strength MS medium lacking plant growth regulators. Flow cytometric analysis revealed that all plantlets obtained were diploid, and thus, appeared to be of embryonic origin. These plantlets were easily acclimatized and were without any morphological abnormalities. The immature seed -derived soft callus lines, thus, may be valuable as an efficient plant regeneration system of this species.
  • Endosperm-derived triploid plant regeneration in diploid Actinidia kolomikta, a cold-hardy kiwifruit relative
    Issei Asakura, Yoichiro Hoshino
    SCIENTIA HORTICULTURAE, 219, 53, 59, ELSEVIER SCIENCE BV, 2017年05月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Endosperm culture is a unique technique for producing a triploid plant from diploid plants. In the present study, endosperm of a diploid cold-hardy kiwifruit relative, Actinidia kolomikta (Maxim. & Rupr.) Maxim., was cultured to regenerate triploid plants and expand ploidy variation in this mostly diploid species. Endosperm derived from fruits harvested 6-10 weeks after flowering (WAF) was used for the initial explants to examine the effect of plant growth regulators on callus formation and organogenesis. The endosperm -derived calli induced in MS medium with above 1.0 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) were soft, small, and translucent, and no morphogenic response was observed. Conversely, the endosperm -derived yellow-green calli induced in the medium with 2,4-D (0.1 mg L-1) plus kinetin (1.0, 2.0, and 5.0 mg L-1) were hard, compact, fast growing, and showed adventitious shoot primordia. These primordia elongated into shoots when the calli were transferred to MS medium supplemented with zeatin (1.0, 2.0, and 4.0 mg L-1) under continuous light conditions. The shoots developed roots on half strength MS medium lacking plant growth regulators. In total, 14 plants were obtained. Flow cytometry and chromosomal analysis confirmed that the regenerated plants were triploid, suggesting that the plant tissue originated from the endosperm. This study reveals that the endosperm of A. kolomikta has plant regeneration abilities and that the regenerated plants have the same ploidy level as the endosperm (triploid). These results may be of use for ploidy manipulation or polyploidy breeding in other species in the genus Actinidia. (C) 2017 Elsevier B.V. All rights reserved.
  • Branch regeneration induced by sever damage in the brown alga Dictyota dichotoma (dictyotales, phaeophyceae)
    Atsuko Tanaka, Yoichiro Hoshino, Chikako Nagasato, Taizo Motomura
    PROTOPLASMA, 254, 3, 1341, 1351, SPRINGER WIEN, 2017年05月, [査読有り]
    英語, 研究論文(学術雑誌), Tissue wounds are mainly caused by herbivory, which is a serious threat for macro-algae, and brown algae are known to regenerate branches or buds in response to wounding. In the present paper, we describe a branch regeneration system, induced by sever damage, in the brown alga Dictyota dichotoma. Segmentations of juvenile thalli induced branch regenerations unless explants possessed apical cells. Apical excisions in distinct positions elucidated that disruption of an apical cell or disconnection of tissue with an apical cell triggered the branch regeneration. Furthermore, spatial positions of regenerated branches seemed to be regulated by the apical region, which was assumed to generate inhibitory effects for lateral branch regeneration. Mechanical incision, which disrupted tissue continuity with the apical region, induced branch regeneration preferentially below the incision. Although we were unable to identify the candidate inhibitory substance, our results suggested that the apical region may have an inhibitory effect on lateral branch regeneration. Additionally, observations of branch regeneration showed that all epidermal cells in D. dichotoma possess the ability to differentiate into apical cells, directly. This may be the first report of algal transdifferentiation during the wound-stress response.
  • Dynamics of nuclear phase changes during pollen tube growth by using in vitro culture in Petunia
    Yoichiro Hoshino, Nozomi Eiraku, Yoshimi Ohata, Fuminori Komai
    SCIENTIA HORTICULTURAE, 210, 143, 149, ELSEVIER SCIENCE BV, 2016年10月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), In flowering plants, pollen germinates to deliver sperm cells to the egg and central cells during double fertilization. In a bicellular pollen species, sperm cells are formed as the pollen tube grows. However, the process of pollen tube growth is difficult to observe as it proceeds in inside the pistil. Here, an in vitro liquid pollen culture system was applied to pollen of Petunia cultivars to analyze the nuclear phase transitions during pollen tube growth by microscopy and flow cytometry. The pollen germination frequencies for eight Petunia cultivars were initially evaluated. Among them, the cultivar 'Chiffon' showed the highest pollen germination frequency and was used for further analysis. Fluorescence microscopy revealed that pollen had two nuclei just after germination, indicating the generative and vegetative cells. Twelve hours after pollen germination, three nuclei were observed in the cultures, suggesting that nuclei indicated two sperm cells and one vegetative cell had developed and the generative cells had divided in vitro. Flow cytometry successfully detected nuclear phase changes during pollen tube growth, and the analysis suggested male germ unit formation, which develops from the connection between the generative and vegetative cells, or between the two sperm cells and the vegetative cell. These results show that the pollen grains could germinate in vitro and generative cells could divide to form sperm cells under these culture conditions. This in vitro liquid pollen culture system will shed light on physiological events regulated in pollen tubes. (C) 2016 Elsevier B.V. All rights reserved.
  • Suppression of B function strongly supports the modified ABCE model in Tricyrtis sp (Liliaceae)
    Masahiro Otani, Ahmad Sharifi, Shosei Kubota, Kanako Oizumi, Fumi Uetake, Masayo Hirai, Yoichiro Hoshino, Akira Kanno, Masaru Nakano
    SCIENTIFIC REPORTS, 6, NATURE PUBLISHING GROUP, 2016年04月, [査読有り]
    英語, 研究論文(学術雑誌), B class MADS-box genes play important roles in petal and stamen development. Some monocotyledonous species, including liliaceous ones, produce flowers with petaloid tepals in whorls 1 and 2. A modified ABCE model has been proposed to explain the molecular mechanism of development of two-layered petaloid tepals. However, direct evidence for this modified ABCE model has not been reported to date. To clarify the molecular mechanism determining the organ identity of two-layered petaloid tepals, we used chimeric repressor gene-silencing technology (CRES-T) to examine the suppression of B function in the liliaceous ornamental Tricyrtis sp. Transgenic plants with suppressed B class genes produced sepaloid tepals in whorls 1 and 2 instead of the petaloid tepals as expected. In addition, the stamens of transgenic plants converted into pistil-like organs with ovule- and stigma-like structures. This report is the first to describe the successful suppression of B function in monocotyledonous species with two-layered petaloid tepals, and the results strongly support the modified ABCE model.
  • Distribution, Ploidy Levels, and Fruit Characteristics of Three Actinidia Species Native to Hokkaido, Japan
    Issei Asakura, Yoichiro Hoshino
    HORTICULTURE JOURNAL, 85, 2, 105, 114, JAPAN SOC HORTICULTURAL SCI, 2016年04月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), The genus Actinidia includes widely-sold kiwifruit, and is thus horticulturally important. We investigated the distribution, ploidy levels, and fruit characteristics of the natural populations of three edible Actinidia species [Actinidia arguta (Siebold & Zucc.) Planch. ex Miq., Actinidia kolomikta (Maxim. & Rupr.) Maxim., and Actinidia polygama (Siebold & Zucc.) Planch. ex Maxim.] in Hokkaido, the northern island of Japan. Actinidia arguta and A. kolomikta were common, and their habitat ranges overlapped. Actinidia polygama was less common, and its habitat was mostly limited to lowland deciduous forests. Flow cytometric analysis revealed that all wild collections of A. kolomikta and A. polygama were diploid, and that A. arguta was tetraploid, suggesting a lack of intraspecific ploidy variation. Fruit shape varied from round to ovoid in A. arguta, ranged from ovoid to ellipsoidal in A. kolomikta, and was ellipsoidal in A. polygama. The fruit skin of all species was glabrous, and skin color was orange in A. polygama, green to dark green in A. kolomikta, and light to dark green in A. arguta. The fresh weight of A. kolomikta fruit was less than that of A. arguta, and the soluble solids content (SSC) of the fruits varied widely within species. One sample of A. arguta had extremely high SSC (average Brix of 30.8%). The ascorbic acid content (AAC) was the highest in A. kolomikta (up to 805 mg per 100 g fresh weight). Actinidia arguta and A. kolomikta germplasm may be useful for breeding new kiwifruit varieties for cultivation in cold-temperate regions.
  • Pollen morphology and its taxonomic significance in the genus Bomarea Mirb. (Alstroemeriaceae) - II. Subgenus Bomarea
    Abul Khayer Mohammad Golam Sarwar, Yoichiro Hoshino, Hajime Araki
    ACTA BOTANICA BRASILICA, 29, 4, 586, 596, SOC BOTANICA BRASIL, 2015年10月, [査読有り]
    英語, 研究論文(学術雑誌), Pollen morphology of 52 species (out of c. 79) of the Bomarea subgenus Bomarea was examined using light microscopy and scanning electron microscopy (SEM), or using SEM alone. The studied species of Bomarea were stenopalynous, characterized by large, oblate, monosulcate monads with reticulate exine sculpture in most species. Wide variation was observed in quantitative palynological features. The studied taxa were divided into four major groups based on exine ornamentation observed under SEM: microreticulate, reticulate, coarsely rugulate, and psilate-perforate. The reticulate exine sculpture may be a plesiomorphic character state for the genus Bomarea, and the coarsely rugulate and finely rugulate-perforate or psilate-perforate exine sculptures may have evolved independently more than once. In agglomerative hierarchical clustering (AHC) analyses of the genus Bomarea using quantitative pollen data, the studied species were distributed in either two (similarity-based) or four (dissimilarity-based) major clusters. Neither the recent molecular phylogenetic analyses nor the AHC analyses of Bomarea have recovered clades/dusters that represent traditionally recognized subgeneric taxa for the genus. Therefore, the most reliable infrageneric classification of Bomarea can be achieved by combining morphological, palynological, and molecular data from more extentive sampling of all the species.
  • Assessment of genetic relationships among cultivated and wild Rubus accessions using AFLP markers
    Tomoya Miyashita, Hisato Kunitake, Norishige Yotsukura, Yoichiro Hoshino
    SCIENTIA HORTICULTURAE, 193, 165, 173, ELSEVIER SCIENCE BV, 2015年09月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), The genus Rubus comprises more than 740 species divided into 12 subgenera and contains the raspberries, blackberries, arctic fruits, and flowering raspberries, all of which have been utilized in breeding programs. The objective of this study was to evaluate the phylogenetic relationships among wild and cultivated Rubus species mainly collected in Japan. To evaluate genetic resources in Rubus, 81 accessions were analyzed with three amplified fragment length polymorphism (AFLP) primer pairs and data were analyzed with the neighbor-joining and unweighted pair group methods with arithmetic mean. Two of the generated phylogenetic trees grouped subgenera Anoplobatus, Eubatus, Idaeobatus, and Malachobatus into different clusters. Accessions of Rubus idaeus L var. aculeatissimus collected from four regions in Hokkaido formed distinct clusters reflecting sampling sites. Four hybrid accessions between raspberry cultivars (R. idaeus L.) and R. idaeus L. var. aculeatissimus, and one hybrid accession between a raspberry cultivar and Rubus spectabilis Pursh were clearly distinguished from parental accessions. These results indicated that AFLP markers are a reliable technique for assessing genetic diversity and studying phylogenetic relationships in Rubus. Data from diversity and phylogenetic studies revealed valuable information on the availability of unique fragments in different accessions that would be useful for the development of improved genotypes through conventional breeding and marker-assisted selection. (C) 2015 Elsevier B.V. All rights reserved.
  • Evidence for a Common Origin of Homomorphic and Heteromorphic Sex Chromosomes in Distinct Spinacia Species
    Satoshi Fujito, Satoshi Takahata, Reimi Suzuki, Yoichiro Hoshino, Nobuko Ohmido, Yasuyuki Onodera
    G3-GENES GENOMES GENETICS, 5, 8, 1663, 1673, GENETICS SOCIETY AMERICA, 2015年08月, [査読有り]
    英語, 研究論文(学術雑誌), The dioecious genus Spinacia is thought to include two wild relatives (S. turkestanica Ilj. and S. tetrandra Stev.) of cultivated spinach (S. oleracea L.). In this study, nuclear and chloroplast sequences from 21 accessions of Spinacia germplasm and six spinach cultivars or lines were subjected to phylogenetic analysis to define the relationships among the three species. Maximum-likelihood sequence analysis suggested that the Spinacia plant samples could be classified into two monophyletic groups (Group 1 and Group 2): Group 1 consisted of all accessions, cultivars, and lines of S. oleracea L. and S. turkestanica Ilj. and two of five S. tetrandra Stev. accessions, whereas Group 2 was composed of the three remaining S. tetrandra Stev. accessions. By using flow cytometry, we detected a distinct difference in nuclear genome size between the groups. Group 2 also was characterized by a sexual dimorphism in inflorescence structure, which was not observed in Group 1. Interspecific crosses between the groups produced hybrids with drastically reduced pollen fertility and showed that the male is the heterogametic sex (XY) in Group 2, as is the case in S. oleracea L. (Group 1). Cytogenetic and DNA marker analyses suggested that Group 1 and Group 2 have homomorphic and heteromorphic sex chromosome pairs (XY), respectively, and that the sex chromosome pairs of the two groups evolved from a common ancestral pair. Our data suggest that the Spinacia genus may serve as a good model for investigation of evolutionary mechanisms underlying the emergence of heteromorphic sex chromosome pairs from ancestral homomorphic pairs.
  • In Vitro Self-incompatible-like Response Applied for Protein Identification and Gene Expression Analysis in Citrus Cultivars, Banpeiyu and Hyuganatsu               
    Yiran Li, Akiha Abe, Takuichi Fuse, Tomonari Hirano, Yoichiro Hoshino, Hisato Kunitake
    Journal of the American Society for Horticultural Science, 140, 4, 339, 345, 2015年07月, [査読有り]
    英語, 研究論文(学術雑誌)
  • Pollen morphology and its taxonomic significance in the genus Bomarea Mirb. (Alstroemeriaceae) - I. Subgenera Baccata, Sphaerine, and Wichuraea
    Abul Khayer Mohammad Golam Sarwar, Yoichiro Hoshino, Hajime Araki
    ACTA BOTANICA BRASILICA, 29, 3, 425, 432, SOC BOTANICA BRASIL, 2015年07月, [査読有り]
    英語, 研究論文(学術雑誌), Pollen morphology of 24 of the 33 species of three Bomarea subgenera, Baccata, Sphaerine, and Wichuraea, was examined by light microscopy (LM) and scanning electron microscopy (SEM), or SEM alone. The studied species of Bomarea were stenopalynous, characterized by large, monosukate monads with reticulate exine sculpture in most species. Opercula-like structures were present on the sulcus in B. huanuco and B. involucrosa. Differences in pollen size, exine thickness, and exine sculpture were observed. The studied taxa were divided into four major groups based on exine ornamentation observed under SEM: microreticulate, reticulate, coarsely rugulate, or finely rugulate-perforate. Pollen characters alone did not appear to correlate clearly with the current subgeneric classification of Bomarea, but they may have some taxonomic utility below the subgeneric level. The most reliable infrageneric classification of Bomarea can be achieved through combined analyses of morphological, palynological, and molecular data from larger samples of specimens of all the species.
  • Evaluation of seedling emergence and relative DNA content under dry soil conditions of wild Festuca arundinacea populations collected in Iran
    Iman Rohollahi, Nayer Azam Khoshkholghsima, Toshihiko Yamada, Mohsen Kafi, Yoichiro Hoshino, Abdolmajid Liaghat, Ali Ashraf Jafari
    GRASSLAND SCIENCE, 61, 1, 6, 14, WILEY-BLACKWELL, 2015年03月, [査読有り]
    英語, 研究論文(学術雑誌), Tall fescue (Festuca arundinacea Schreb.) is an important forage and turf grass species adapted to cold, arid and semiarid environments. Germination and seedling establishment of F.arundinacea cultivars is constrained by the low soil moisture found in these regions. Genome size was found to positively correlate with latitude of origin, suggesting that structural changes in DNA may play a role in environmental adaptation. We investigated the effects of low soil moisture on the germination and early establishment and their correlation with relative DNA content using 14 wild F.arundinacea populations collected from various regions in Iran and two commercial turf cultivars. The populations were evaluated under 100% (-0.03 matric potential [MPa]), 80% (-0.2), 60% (-0.6) and 40% (-1.4) field capacity (FC) conditions. Seed germination and root and leaf growth decreased under reduced soil water content. Cluster analysis revealed that the populations fell into four groups. Populations in cluster I showed the greatest tolerance to low soil moisture and cluster IV was the least tolerant. Isfahan' and Gonabad' populations had the best final germination and longer leaf and root length at 40% FC. Studies to identify indices that were involved in drought resistance revealed that final germination, leaf length and seedling vigor index were most important evaluating indicators for F.arundinacea populations. Flow cytometric relative DNA content of the wild populations was negatively correlated with germination, leaf length and seedling vigor index. The drought tolerance populations in cluster I could be potentially useful germplasm for a breeding program to develop superior cultivars for arid and semiarid regions.
  • In vitro strategies for the conservation of some medicinal and horticultural climbers
    T. Dennis Thomas, Yoichiro Hoshino
    Biotechnological Strategies for the Conservation of Medicinal and Ornamental Climbers, 259, 290, Springer International Publishing, 2015年01月01日, [査読有り]
    英語, 論文集(書籍)内論文, There has been an increase in use of traditional medicine during the last decade due to the reason that medicinal plant-derived drugs are more effective against several diseases as compared to antibiotics and other medicines and the former has less side effects on patients. Moreover, medicinal plants are a rich source of antibacterial compounds. The increased use and harvest of medicinal plants coupled with habitat destruction caused a decrease in natural population of most of the medicinal plants. In vitro conservation strategies can not only offer large number of plants in limited time but also it can decrease tension on natural populations. Similarly, several horticultural plants including vegetable crops need improvements and therefore in vitro conservation strategies could be applied for their further progress. The present review is an attempt to provide information on various in vitro conservation strategies adopted in selected medicinal and horticultural climbers. The important medicinal and horticultural climbers such as Benincasa hispida, Cardiospermum halicacabum, Tylophora indica, Sarcostemma brevistigma, Cyclea peltata, Caesalpinia bonduc, Clitoria ternatea, Hemidesmus indicus, Momordica charantia, and Embelia ribes have been discussed in detail.
  • Interploid and intraploid hybridizations to produce polyploid Haskap (Lonicera caerulea var. emphyllocalyx) plants
    Tomomi Miyashita, Yoichiro Hoshino
    EUPHYTICA, 201, 1, 15, 27, SPRINGER, 2015年01月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), We produced polyploid Haskap (Lonicera caerulea var. emphyllocalyx) plants by performing interploid and intraploid crosses of wild accessions. Embryo rescue in a tetraploid (4x) x diploid (2x) cross produced triploid plants; reciprocal 2x x 4x cross failed to produce viable seeds. Intraploidy crosses of 4x x 4x produced mostly tetraploids but also several hexaploid (6x) and octoploid (8x) plants. Using hexaploids obtained from this cross, we examined reciprocal 4x-6x crosses and found that both produced pentaploid plants. An octoploid was produced by applying colchicine to a tetraploid; a 4x x 8x cross using this plant and aided by embryo rescue culture produced three hexaploid plants, with an aneuploid number of chromosomes. Several plants obtained in this study flowered and set fruits. We discuss the overall efficiency of producing polyploid plants in interploid and intraploid crosses.
  • Characterization of CYCLOIDEA-like genes in controlling floral zygomorphy in the monocotyledon Alstroemeria
    Yoichiro Hoshino, Toshiya Igarashi, Masumi Ohshima, Koichi Shinoda, Naho Murata, Akira Kanno, Masaru Nakano
    SCIENTIA HORTICULTURAE, 169, 6, 13, ELSEVIER SCIENCE BV, 2014年04月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), The CYCLOIDEA (CYC) gene controls the development of zygomorphic flowers and the determination of adaxial identity of floral organs in the model developmental system of Antirrhinum majus. However, whether CYC homologue genes also control floral zygomorphy in monocotyledon Alstroemeria plants is yet unknown. In this study, we investigated CYC-like genes in the monocotyledons Alstroemeria aurea, Alstroemeria magenta, and Alstroemeria pelegrina var. rosea, all of which have zygomorphic flowers. Since the CYC gene belongs to the T-complex protein (TCP) gene family of transcription factors, cloning of CYC-like sequences was performed using rapid amplification of cDNA ends (RACE)-polymerase chain reaction (PCR) by using degenerate primers designed for the TCP domain. We cloned 1 CYC-like sequence each from A. aurea (AaTCP1, accession number AB714967 in the GenBank/EMBL/DDBJ databases) and A. magenta (AmTCP1, AB714970), and two CYC-like sequences from A. pelegrina var. rosea (ApTCP1, AB714968; and ApTCP2, AB714969). The deduced amino acid sequences of AaTCP1, AmTCP1, ApTCP1, and ApTCP2 shared 67.7%, 67.7%, 71.0%, and 64.5% identities, respectively, with the TCP domain in CYC. Molecular phylogenetic analysis indicated that three CYC-like genes from Alstroemeria belonged to the ZinTBL1b clade in the CYC-/tbl -like subfamily. Reverse transcription (RT)-PCR and in situ hybridization analyses showed that AaTCP1 transcripts were specifically detected in flower buds and localized in the base of adaxial inner perianth of A. aurea. These results suggest that CYC-like genes are also involved in the development of floral asymmetry and the determination of adaxial identity of floral organs in the monocotyledon Alstroemeria. (C) 2014 Elsevier B.V. All rights reserved.
  • MicroRNA319 Positively Regulates Cold Tolerance by Targeting OsPCF6 and OsTCP21 in Rice (Oryza sativa L.)
    Sun-ting Wang, Xiao-li Sun, Yoichiro Hoshino, Yang Yu, Bei Jia, Zhong-wen Sun, Ming-zhe Sun, Xiang-bo Duan, Yan-ming Zhu
    PLOS ONE, 9, 3, PUBLIC LIBRARY SCIENCE, 2014年03月, [査読有り]
    英語, 研究論文(学術雑誌), The microRNA319 (miR319) family is conserved among diverse plant species. In rice (Oryza sativa L.), the miR319 gene family is comprised of two members, Osa-miR319a and Osa-miR319b. We found that overexpressing Osa-miR319b in rice resulted in wider leaf blades and delayed development. Here, we focused on the biological function and potential molecular mechanism of the Osa-miR319b gene in response to cold stress in rice. The expression of Osa-miR319b was down-regulated by cold stress, and the overexpression of Osa-miR319b led to an enhanced tolerance to cold stress, as evidenced by higher survival rates and proline content. Also, the expression of a handful of cold stress responsive genes, such as DREB1A/B/C, DREB2A, TPP1/2, was increased in Osa-miR319b transgenic lines. Furthermore, we demonstrated the nuclear localization of the transcription factors, OsPCF6 and OsTCP21, which may be Osa-miR319b-targeted genes. We also showed that OsPCF6 and OsTCP21 expression was largely induced by cold stress, and the degree of induction was obviously repressed in plants overexpressing Osa-miR319b. As expected, the down-regulation of OsPCF6 and OsTCP21 resulted in enhanced tolerance to cold stress, partially by modifying active oxygen scavenging. Taken together, our findings suggest that Osa-miR319b plays an important role in plant response to cold stress, maybe by targeting OsPCF6 and OsTCP21.
  • Effects of different wavelengths of LED light on pollen germination and direction of pollen tube elongation in Cyrtanthus mackenii
    Yuri Hoyo, Kazuhiro Fujiwara, Yoichiro Hoshino
    Advances in Horticultural Science, 28, 4, 190, 194, 2014年, [査読有り]
  • DNA damage response in male gametes of Cyrtanthus mackenii during pollen tube growth
    T. Hirano, K. Takagi, Y. Hoshino, T. Abe
    AoB Plants, 5, 0, plt004, plt004, Oxford University Press (OUP), 2013年02月27日, [査読有り]
    研究論文(学術雑誌)
  • In vitro culture of endosperm and its application in plant breeding: Approaches to polyploidy breeding
    Yoichiro Hoshino, Tomomi Miyashita, Thuruthiyil Dennis Thomas
    SCIENTIA HORTICULTURAE, 130, 1, 1, 8, ELSEVIER SCIENCE BV, 2011年08月, [査読有り]
    英語, This review article provides an overview of plant regeneration from endosperm to produce polyploid plants. We discuss the endosperm-ploidy levels and its applications in plant breeding. Representative cases of successful endosperm culture and prospects of endosperm culture are described. In the first part of the review, i.e., the background of endosperm culture, we discuss the importance of triploid plants, meaning plants derived from the endosperm of diploid plants. A brief history of endosperm culture is also provided, and a description of plant regeneration systems from the endosperm is given. The influence of culture medium on callus induction and regeneration is indicated. Finally, prospects for endosperm culture are proposed, and novel approaches for polyploidy breeding using endosperm culture are described. (C) 2011 Elsevier B.V. All rights reserved.
  • Establishment of an efficient in vitro culture and particle bombardment-mediated transformation systems in Miscanthus sinensis Anderss., a potential bioenergy crop
    Xun Wang, Tetsuya Yamada, Fan-Jiang Kong, Yuki Abe, Yoichiro Hoshino, Hiroko Sato, Tadashi Takamizo, Akira Kanazawa, Toshihiko Yamada
    GLOBAL CHANGE BIOLOGY BIOENERGY, 3, 4, 322, 332, WILEY-BLACKWELL, 2011年08月, [査読有り]
    英語, 研究論文(学術雑誌), Plants belonging to the genus Miscanthus are considered promising bioenergy crops. Here, we report the establishment of tissue culture system through particle bombardment-mediated transformation in Miscanthus sinensis Anderss. Callus was induced efficiently from mature seeds in a medium containing a combination of a relatively high level of 2,4-dichlorophenoxyacetic acid (5 mg L-1) and a relatively low level of 6-benzyladenine (BA) (0.01 mg L-1). Callus induction potential of 18 accessions of M. sinensis, which were collected from various sites in Japan, was compared. Significant correlation was detected between compact (embryogenic) callus induction frequency and average annual air temperature in collection sites. An accession from Tanegashima Island showed the highest production of compact callus. We found that a higher level of BA causes callus browning; the 2 mg L-1 BA is the optimal concentration for regeneration. Both compact and friable calli were suitable for particle bombardment transformation. Through selection under the presence of 50 mg L-1 hygromycin for 3 weeks and further selection under the presence of 150 mg L-1 for 1 month, hygromycin-resistant calli survived, of which 72.2% had been entirely transformed. Plants were regenerated from calli in the presence of hygromycin; transcripts of the hpt and gfp genes, which were cobombarded to the calli, were detected in the regenerated plants. This is the first report on the establishment of the in vitro culture of M. sinensis using mature seeds, the variation of callus formation among accessions collected from various sites in Japan, and particle bombardment-mediated transformation in the genus Miscanthus.
  • Optimizing embryo and shoot tip derived callus production and high frequency plant regeneration in the model grass Brachypodium distachyon (L.) P. Beauv
    T. Dennis Thomas, Hoshino Yoichiro
    PLANT BIOSYSTEMS, 145, 4, 924, 930, TAYLOR & FRANCIS LTD, 2011年, [査読有り]
    英語, 研究論文(学術雑誌), Brachypodium distachyon has been recently proposed as a new model system for monocot functional genomics because of its unique properties such as small size, short generation time, self-fertile, chromosome base number five and small genome size. In this study, we have standardized a protocol to obtain 100% green plant regeneration from embryo and shoot tip explants. The optimum callus induction in terms of percentage of cultures responding and callus growth was observed on Murashige and Skoog (MS) medium supplemented with 4 mg/l 2, 4 dichlorophenoxy acetic acid (2, 4-D) and 0.5 mg/l kinetin (Kn). On this medium, 88% and 100% explants responded from embryo and shoot tip explants, respectively. Comparatively, shoot tip explants produced better response than embryo explants. The callus multiplied and maintained on MS medium supplemented with 2, 4-D (4 mg/l) and Kn (0.5 mg/l) for about 10 months without decline in growth rate. The callus subcultured on MS medium supplemented with thidiazuron (TDZ; 0.5-7 mg/l) or 6-benzyladenine (BA; 0.5-7 mg/l) alone or in combination with naphthalene acetic acid (NAA; 0.5 mg/l) for regeneration. Maximum shoot regeneration was observed on MS medium supplemented with 5 mg/l TDZ and 0.5 mg/l NAA. On this medium, 87% and 96% cultures responded with mean shoot number 10.2 and 16.6 from embryo and shoot derived callus, respectively. The shoots were rooted on MS medium supplemented with either NAA (2-8 mg/l) or indole-3-butyric acid (IBA; 2-8 mg/l). The optimum response (98%) was observed on MS medium supplemented with 4 mg/l IBA with 4.5 roots per shoot. The rooted plantlets were successfully transplanted with 90% success. The acclimatized plants produced flowers and viable seeds.
  • Ploidy distribution and DNA content variations of Lonicera caerulea (Caprifoliaceae) in Japan
    Tomomi Miyashita, Hajime Araki, Yoichiro Hoshino
    JOURNAL OF PLANT RESEARCH, 124, 1, 1, 9, SPRINGER TOKYO, 2011年01月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Ploidy level and geographical distribution were investigated in Japanese Lonicera caerulea L. Flow cytometric analysis revealed the presence of DNA diploid and DNA tetraploid plants in Japan. Chromosome observation confirmed that diploid and tetraploid plants showed 2n = 2x = 18 and 2n = 4x = 36, respectively. The DNA diploid populations were found only in lowland mires, Betsukai, Bekanbeushi, Kushiro and Kiritappu located in eastern Hokkaido. On the other hand, DNA tetraploid populations were distributed in a wide area of Hokkaido, and mainland of Japan. The habitats of DNA tetraploid plants were lowland to alpine region. The DNA content measurement with flow cytometry revealed significant differences in the relative DNA contents among DNA tetraploid populations. The relative DNA content within DNA tetraploid populations varied 1.157-fold at maximum, and might correlate with altitude indicating that DNA contents were smaller as altitude increases. The wide area of distribution in various environments of DNA tetraploid plants suggested the adaptability of the tetraploid plants. Although diploid and tetraploid populations were found, no triploid was detected, indicating crossing difficulty between diploid and tetraploid as confirmed by crossing experiment.
  • Evaluation of the horticultural traits of genus Alstroemeria and genus Bomarea (Alstroemeriaceae)
    Yukiko Kashiharai, Koichi Shinoda, Naho Murata, Hajime Araki, Yoichiro Hoshino
    TURKISH JOURNAL OF BOTANY, 35, 2, 239, 245, TUBITAK SCIENTIFIC & TECHNICAL RESEARCH COUNCIL TURKEY, 2011年, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), We investigated the flower characteristics of 7 Alstroemeriaceae plants - 5 Alstroemeria L. species and 2 Bomarea Mirb. species. The flower and tepal sizes, and tepal colours were compared. The flower shape in Alstroemeria psittacina Lehm. was found to be similar to that in Bomarea coccinea (Ruiz & Pay.) Baker. The length/width ratio of B. salsilla (L.) Mirb. was intermediate compared with that of the other species. These preliminary data will be useful in selecting wild species in order to examine interspecific or intergeneric hybridisations in the breeding of Alstroemeriaceae plants.
  • Rapid and efficient callus induction and plant regeneration from seeds of zoysiagrass (Zoysia japonica Steud.)
    Xun Wang, Yoichiro Hoshino, Toshihiko Yamada
    GRASSLAND SCIENCE, 56, 4, 198, 204, WILEY-BLACKWELL PUBLISHING, INC, 2010年12月, [査読有り]
    英語, 研究論文(学術雑誌), A rapid and efficient in vitro culture system has been established in zoysiagrass (Zoysia japonica Steud.). Embryos isolated from mature seeds were used to induce callus, which improved callus formation and shortened the culture period. A high concentration of 2,4-dichlorophenoxyacetic acid (5 mg L-1) in combination with low concentration of 6-benzyladenine (0.2 mg L-1) was proved to be suitable for callus induction in zoysiagrass. Eight types of calli were observed; friable callus was the most regenerable type. Shoot regeneration efficiency was improved by using 1 mg L-1 of thidiazuron with 1-naphthaleneacetic acid (0.05 mg L-1) and gibberellic acid (0.1 mg L-1). Zoysiagrass shoots continued to grow in MS medium, and rooted in half-strength MS medium. With this system, it appears the duration of in vitro regeneration could be shortened to 16 weeks for zoysiagrass, i.e. from inoculation of embryos to regeneration of plantlets with shoots and roots.
  • In vitro propagation for the conservation of a rare medicinal plant Justicia gendarussa Burm. f. by nodal explants and shoot regeneration from callus
    T. Dennis Thomas, Hoshino Yoichiro
    ACTA PHYSIOLOGIAE PLANTARUM, 32, 5, 943, 950, SPRINGER HEIDELBERG, 2010年09月, [査読有り]
    英語, 研究論文(学術雑誌), Justicia gendarussa is a valuable medicinal plant and various parts of this plant are pharmaceutically used for the treatment of different diseases. In vitro regeneration of shoot buds was obtained from culture of nodal cuttings as well as shoot regeneration from callus. The nodal cuttings differed in shoot proliferation in terms of percentage of explants that responded and average shoot length with various concentrations (4.4, 8.9, 13.3, 17.7, 22.2 mu M) of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron. In all treatments, one shoot was invariably present. Optimum 87% of cultures responded with an average shoot length of 4.4 cm on Murashige and Skoog (MS) medium supplemented with 17.7 mu M BA. Callus was induced from the mature leaf segments on MS medium supplemented with Kn (4.7, 13.9, 23.2 mu M) alone or in combination with 2, 4-dichlorophenoxyacetic acid (2, 4-D; 2.3 mu M, 4.5 mu M). Optimum callus induction (78%) was obtained on MS medium supplemented with 14 mu M Kn and 4.5 mu M 2, 4-D. When the callus was subcultured on MS medium fortified with BA (8.9, 17.7, 26.6 mu M) or Kn (9.3, 18.6, 27.9 mu M) alone or in combination with a naphthalene acetic acid (NAA; 2.7, 5.4 mu M), shoot regeneration was obtained. The highest response (92%) was observed on MS medium containing 17.7 mu M BA and 5.4 lM NAA. On this medium, an average number of 12.2 shoots were obtained per responding callus. The shoots obtained from callus and nodal cuttings were rooted with a frequency of 73% on MS medium augmented with 9.8 lM indole-3-butyric acid. The rooted shoots were successfully transplanted to soil and sand mixture (1: 1) with 90% survival rate. The protocol standardized for shoot proliferation and regeneration in J. gendarussa from nodal cuttings and leaf-derived callus is suitable for micropropagation and conservation of this essential medicinal plant.
  • Interspecific hybridization in Lonicera caerulea and Lonicera gracilipes: The occurrence of green/albino plants by reciprocal crossing
    Tomomi Miyashita, Yoichiro Hoshino
    SCIENTIA HORTICULTURAE, 125, 4, 692, 699, ELSEVIER SCIENCE BV, 2010年07月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Lonicera caerulea L. var. emphyllocalyx (Maxim.) Nakai is a berry crop cultivated in cold regions. So far, commercial cultivars have been mainly introduced from selection of wild plants. Therefore, fruit traits and other agricultural characteristics have been limited. In this study, interspecific crosses between L. caerulea var. emphyllocalyx and Lonicera gracilipes var. glabra Miguel were examined to increase genetic variability of L. caerulea var. emphyllocalyx. Seedlings were obtained from reciprocal crosses between L caerulea var. emphyllocalyx and L. gracilipes var. glabra. The hybrid nature of seedlings was confirmed with random amplified polymorphic DNA analysis. Viable green plants were obtained efficiently from L gracilipes var. glabra x L. caerulea var. emphyllocalyx. In contrast, all plants produced from L caerulea var. emphyllocalyx x L. gracilipes var. glabra were albino. These albino plants were very weak and only survived in culture condition. The chlorophyll deficiency was unilaterally observed, suggesting the occurrence of nuclear-cytoplasmic incompatibility. Viable F(I) hybrids obtained from L gracilipes var. glabra x L caerulea var. emphyllocalyx are amphidiploid (2n = 4x = 36) as showing same to both parents. The hybrid plants are expected to increase the variability of fruit traits, and may have heat tolerance from L gracilipes var. glabra. (C) 2010 Elsevier B.V. All rights reserved.
  • Sperm dimorphism in terms of nuclear shape and microtubule accumulation in Cyrtanthus mackenii
    Tomonari Hirano, Yoichiro Hoshino
    SEXUAL PLANT REPRODUCTION, 23, 2, 153, 162, SPRINGER, 2010年06月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Pollen tubes of Cyrtanthus mackenii, a species with bicellular pollen, were cultured in vitro to investigate nuclear phase changes during generative cell division and male germ unit (MGU) formation, using flow cytometric analysis. Results revealed that sperm cells were formed after 12 h of culture. During sperm maturation, the nuclei of sperm cells were not associated with the vegetative nucleus (unassociated sperm cells; Sua) and became longer than those of sperm cells associated with the vegetative nucleus (Svn). These findings indicate that the pair of sperm cells in the C. mackenii MGU is dimorphic in terms of nuclear shape. Dimorphism coincides with anti-alpha-tubulin antibody immunofluorescence, which was higher in the Sua than in Svn. Following treatment with oryzalin, triggering microtubule depolymerization, differences between nuclear shapes in the two sperm nuclei disappeared, suggesting that microtubule accumulation between sperm cells in the MGU correlates with differences in the nuclear shape.
  • Pollen morphology and infrageneric classification of Alstroemeria L. (Alstroemeriaceae)
    Abul Khayer Mohammad Golam Sarwar, Yoichiro Hoshino, Hajime Araki
    GRANA, 49, 4, 227, 242, TAYLOR & FRANCIS AS, 2010年, [査読有り]
    英語, 研究論文(学術雑誌), Pollen morphology of 53 taxa of the South American genus Alstroemeria was examined using light and scanning electron microscopy (LM and SEM, respectively), or SEM alone, in search of new characters that might contribute to infrageneric classification of the genus. The Alstroemeria species are stenopalynous and characterised by monosulcate and large monads with striate-reticulate exine sculpture. Pollen with auriculae-like structures at each end of the sulcus is reported for the first time in Alstroemeria. Based on exine sculpture, the studied taxa were divided into two major groups; one with striate-reticulate exine (Type I) and the other with finely to coarsely rugulate-psilate exine (Type II). Type I exine sculpture can be further divided into two sub-types based on the width of the muri. Although species from Brazil show relatively closer positions in principal component analysis of quantitative data, agglomerative hierarchical clustering of palynological features does not support a subdivision of this genus into a Brazilian and a Chilean species group. The infrageneric classification of Alstroemeria can be reliably achieved by combined analyses of morphological, palynological and molecular data from a larger number of specimens of all species.
  • Plant regeneration with maintenance of the endosperm ploidy level by endosperm culture in Lonicera caerulea var. emphyllocalyx
    Tomomi Miyashita, Takafumi Ohashi, Fukashi Shibata, Hajime Araki, Yoichiro Hoshino
    PLANT CELL TISSUE AND ORGAN CULTURE, 98, 3, 291, 301, SPRINGER, 2009年09月, [査読有り]
    英語, 研究論文(学術雑誌), An endosperm culture of Haskap (Lonicera caerulea var. emphyllocalyx) was established to develop polyploid plants and investigate the regeneration ability of the endosperm. Based on histological analysis of embryo and endosperm development, endosperms at the globular to early torpedo-stages of developing embryos were used to initiate an endosperm culture. Formation of shoot primordia was observed on Murashige and Skoog (MS) medium (Physiol Plant 15:473-497, 1962) containing benzyladenine and indole-3-butyric acid. Shoot primordium formation was confirmed in some genotypes with regeneration frequencies ranging between 1.9 and 10.0%. These proliferated on A1/2 MS medium containing 2.89 mu M gibberellic acid (GA(3)), and then elongated and rooted on MS medium containing 0.44 mu M BA and 2.89 mu M GA(3). These shoots developed into plantlets on A1/2 MS medium. Plantlets maintained ploidy of the endosperm following flow cytometric analysis, thus confirming that these were derived from the endosperm. These results indicated that endosperms were capable of regeneration.
  • Morphological dormancy in seeds of the autumn-germinating shrub Lonicera caerulea var. emphyllocalyx (Caprifoliaceae)
    Shyam S. Phartyal, Tetsuya Kondo, Yoichiro Hoshino, Carol C. Baskin, Jerry M. Baskin
    PLANT SPECIES BIOLOGY, 24, 1, 20, 26, WILEY-BLACKWELL PUBLISHING, INC, 2009年04月, [査読有り]
    英語, 研究論文(学術雑誌), To better understand the germination ecophysiology of the genus Lonicera, the dormancy class, temperature requirements for embryo growth and radicle emergence and phenology of seedling emergence were determined for Lonicera caerulea var. emphyllocalyx. At maturity, seeds have an underdeveloped embryo (approximately 28% of the length of full-grown embryos). Embryos in fresh seeds grew to full length at 15, 20, 20/10 and 25/15 degrees C within 3 weeks, but failed to grow at <= 10 degrees C and at 30 degrees C. Radicles emerged from 86-100% of freshly matured seeds in light at 15, 20, 20/10 and 25/15 degrees C within 28 days, but failed to emerge at 10 degrees C. Radicles emerged equally well in a 12 h photoperiod and in continuous darkness at 25/15 degrees C. Rapid embryo growth and germination over a range of conditions indicate that seeds of this taxon have morphological dormancy (MD); this is the first report of MD in a species of Lonicera. Seeds are dispersed in summer, at which time high temperatures promote embryo growth. Embryos grow to the critical length for germination in approximately 1 month; the peak of seedling emergence occurs in early autumn. Radicles emerged within 2 months from 98% of seeds buried at soil depths of 2 cm and 10 cm in the field in August in Sapporo, Japan; thus, seeds have no potential to form a persistent soil seed bank. However, seeds sown too late in autumn for embryos to grow remained viable and germinated the following summer when temperatures were high enough to promote embryo growth.
  • Detection of changes in the nuclear phase and evaluation of male germ units by flow cytometry during in vitro pollen tube growth in Alstroemeria aurea
    Tomonari Hirano, Yoichiro Hoshino
    JOURNAL OF PLANT RESEARCH, 122, 2, 225, 234, SPRINGER TOKYO, 2009年03月, [査読有り]
    英語, 研究論文(学術雑誌), This study aimed to analyze male gamete behavior from mature pollen to pollen tube growth in the bicellular pollen species Alstroemeria aurea. For mature pollen, pollen protoplasts were examined using flow cytometry. The protoplasts showed two peaks of DNA content at 1C and 1.90C. Flow cytometry at different developmental stages of pollen tubes cultured in vitro revealed changes in the nuclear phase at 9 and 18 h after culture. Sperm cell formation occurred at 6-9 h after culture, indicating that the first change was due to the division of the generative cells into spermcells. After sperm cell formation, the number of vegetative nucleus associations with sperm cells showed a tendency to increase. This association was suggested as the male germ unit (MGU). When sperm cells, vegetative nuclei, and partial MGUs were collected separately from pollen tubes cultured for 18 h and analyzed using a flow cytometer, the sperm cells and vegetative nuclei contained 1C DNA, while the DNA content of partial MGUs was counted as 2C. Therefore, the second change in the nuclear phase, which results in an increase in 2C nuclei, is possibly related to the formation of MGUs.
  • Plant regeneration from suspension cells induced from hypocotyls derived from interspecific cross Alstroemeria pelegrina x A-magenta and transformation with Agrobacterium tumefaciens
    Yoichiro Hoshino, Yukiko Kashihara, Tomonari Hirano, Naho Murata, Koichi Shinoda
    PLANT CELL TISSUE AND ORGAN CULTURE, 94, 1, 45, 54, SPRINGER, 2008年07月, [査読有り]
    英語, 研究論文(学術雑誌), Embryogenic cell suspension cultures were established using the ovule culture of an interspecific cross, Alstroemeria pelegrina var. rosea x A. magenta. Ovules harvested 14 days after pollination were cultured on Murashige and Skoog (MS) medium without plant growth regulators (PGRs); calli were produced on the hypocotyl surface in germinating zygotic embryos. Suspension cells were induced from the calli by using liquid MS media containing 2,4-dichlorophenoxyacetic acid or 4-amino-3,5,6-trichloropyridine-2-carboxylic acid (picloram). Adventitious embryos developed from the suspension cells on half-strength MS medium supplemented with 0.5 mg l(-1) of both alpha-naphthaleneacetic acid and N(6)-benzylaminopurine; they grew into plantlets on the same medium. The plantlets formed rhizomes following transfer to half-strength MS medium without PGRs, and acclimatized plants were easily established. Subsequently, Agrobacterium-mediated transformation system was applied. The suspension cells were co-cultivated with A. tumefaciens strain EHA101/pIG121Hm or LBA4404/pTOK233, both of which contain neomycin phosphotransferase II, hygromycin phosphotransferase and intron-containing beta-glucuronidase (intron-GUS) genes. Seven days after co-cultivation, the cells were subjected to GUS assay; staining was most pronounced in the cells subcultured in a picloram-containing liquid medium and co-cultivated with EHA101/pIG121Hm. The co-cultivated cells were transferred to the MS medium containing picloram and 20 mg l(-1) hygromycin; 1 month later, several hygromycin-resistant callus lines showing GUS activity were obtained. Transgenic plants were obtained through our plant regeneration system, and foreign gene insertion into the regenerated plants was confirmed by polymerase chain reaction.
  • Flower form alteration by genetic transformation with the class B MADS-box genes of Agapanthus praecox spp. orientalis in transgenic dicot and monocot plants
    Masaru Nakano, Hiroto Umehara, Yoshihiro Hara, Motohide Makino, Mika Igarashi, Mutsumi Nakada, Toru Nakamura, Yoichiro Hoshino, Akira Kanno
    MOLECULAR BREEDING, 20, 4, 425, 429, SPRINGER, 2007年11月, [査読有り]
    英語, 研究論文(学術雑誌), The class B genes, which belong to the MADS-box gene family, play important roles in regulating petal and stamen development in flowering plants. These genes exist in two different types termed DEF- and GLO-like genes, and the B-function is provided by heterodimers of a DEF- and a GLO-like gene product. In the present study, dicot (tobacco and lettuce) and monocot (Tricyrtis hirta) plants were transformed with the GLO-like gene of Agapanthus praecox ssp. orientalis ApGLO alone or in combination with the DEF-like gene of the same plant ApDEF. In two out of 10 transgenic tobacco plants containing ApGLO, sepals partially converted into petaloid organs. For lettuce, ray florets of four out of nine transgenic plants containing ApGLO also developed additional petaloid organs. In two out of five transgenic T. hirta plants containing both ApGLO and ApDEF, organs developed in whorl 4 showed noticeable morphological alteration: they were much longer compared with carpels of non-transgenic plants, and had purple spots overall on the surface as filaments of non-transgenic plants. No morphological alterations were observed in vegetative organs between transgenic and non-transgenic plants for all the three species. The results obtained in the present study indicate a possibility of molecular breeding for flower form alteration by genetic transformation with the class B MADS-box gene(s) of heterologous plant species.
  • Isolation of individual egg cells and zygotes in Alstroemeria followed by manual selection with a microcapillary-connected micropump
    Yoichiro Hoshino, Naho Murata, Koichi Shinoda
    ANNALS OF BOTANY, 97, 6, 1139, 1144, OXFORD UNIV PRESS, 2006年06月, [査読有り]
    英語, 研究論文(学術雑誌), center dot Aims To develop a procedure for isolating living egg cells and zygotes from Alstroemeria ovules.
    center dot Scope An attempt was made to isolate egg cells and zygotes from the ovules of Alstroemeria aurea. The ovules were histologically observed using a clearing procedure which revealed the localization and sizes of the embryo sacs and egg apparatus within the ovules. For the isolation of egg cells, ovules were cut into sections with a surgical blade and treated with an enzyme solution. Subsequently, these ovule sections were dissected using a glass needle under an inverted microscope. Egg cells successfully isolated by this procedure were collected using microcapillaries connected to a micropump. For zygote isolation, ovules were excised from ovaries 24h after self-pollination. By treating excised ovules with an enzyme solution and subsequently dissecting them using a glass needle, zygotes were successfully isolated from the ovules and collected with a microcapillary. The isolated zygotes were associated with pollen tubes and one of the synergids. Egg cells and zygotes were viable for up to 2 h following isolation, as determined by fluorescein diacetate staining.
    center dot Conclusions The procedures for isolating egg cells and zygotes in Alstroemeria were established, and each egg cell and zygote was captured with a microcapillary.
  • Fertilization-induced changes in the microtubular architecture of the maize egg cell and zygote-an immunocytochemical approach adapted to single cells
    Y Hoshino, S Scholten, P von Wiegen, H Lorz, E Kranz
    SEXUAL PLANT REPRODUCTION, 17, 2, 89, 95, SPRINGER, 2004年07月, [査読有り]
    英語, 研究論文(学術雑誌), By using single cell micromanipulation techniques, we developed an immunocytochemical procedure to examine subcellular protein localization in isolated and cultured cells. Localization of microtubules was examined in isolated single egg cells and developing zygotes of maize with anti-alpha-tubulin antibodies. In egg cells, a few cortical microtubules were detected but well organized microtubules were rarely observed. In contrast, distinct cortical microtubules and strands of cytoplasmic microtubules radiating from the nucleus to the cell periphery were observed in developing zygotes. Solely cortical microtubules were observed in zygotes up to 7 h after in vitro fertilization. After this time, radiating microtubules additionally appeared, and persisted during zygote development. These results indicate early and pronounced fertilization-induced changes in microtubular organization in the fertilized egg cell of maize.
  • Horticultural characterization of Angelonia salicariifolia plants transformed with wild-type strains of Agrobacterium rhizogenes
    Y Koike, Y Hoshino, M Mii, M Nakano
    PLANT CELL REPORTS, 21, 10, 981, 987, SPRINGER-VERLAG, 2003年06月, [査読有り]
    英語, 研究論文(学術雑誌), Genetic transformation was carried out with wild-type strains of Agrobacterium rhizogenes for introducing a dwarf trait into the Scrophulariaceous ornamental plant, angelonia (Angelonia salicariifolia). Leaf segments of two angelonia genotypes (Ang.1 and Ang.2) were co-cultivated with mikimopine-type strains of A. rhizogenes. Adventitious roots that showed vigorous growth and increased lateral branching when cultured on half-strength Murashige and Skoog's (MS) basal salts medium lacking plant growth regulators (PGRs) after co-cultivation were selected as putatively transformed lines. All of these selected lines produced mikimopine. Adventitious shoots were efficiently induced from putatively transformed root segments on half-strength MS basal salts medium containing 1 mg l(-1) benzyladenine (BA) under continuous illumination (24-h photoperiod), and the shoots easily rooted following their transfer to half-strength MS basal salts medium lacking PGRs. The transgenic nature of regenerated plants was confirmed by Southern hybridization. Transformed plants frequently died during their acclimatization, and acclimatized plants of eight transformed lines grew very slowly for 1-5 months after transplantation to the greenhouse. Plants of two transformed lines of Ang.2 flowered 4-6 months after transplantation. These transformed plants exhibited phenotypic alterations such as dwarfness and smaller leaves. There were no apparent alterations observed in the number, shape, and size of the flowers. Pollen fertility of the transformed plants was 60-80% based on aceto-carmine staining. These results indicate the possibility of applying A. rhizogenes-mediated transformation for introducing a dwarf trait into angelonia.
  • Explants of Ri-transformed hairy roots of spinach can develop embryogenic calli in the absence of gibberellic acid, an essential growth regulator for induction of embryogenesis from nontransformed roots
    T Ishizaki, Y Hoshino, K Masuda, K Oosawa
    PLANT SCIENCE, 163, 2, 223, 231, ELSEVIER SCI IRELAND LTD, 2002年08月, [査読有り]
    英語, 研究論文(学術雑誌), Hairy roots of spinach (Spinacia oleracea L.) were obtained by inoculation of cotyledon explants with wild-type Agrobacterium rhizogenes A13. Integration of the transfer DNA (T-DNA) of a root-inducing (Ri) plasmid of A. rhizogenes into the plant DNA was confirmed by the polymerase chain reaction. Somatic embryogenesis from explants of hairy roots were induced even when the callus-induction (CI) medium did not contain gibberellic acid (GA(3)) which is considered to be a key growth regulator that is critical for induction of somatic embryogenesis in spinach. Proteins extracted from hairy root-derived calli and from non-transformed root-derived calli generated on medium that contained 10 muM naphthaleneacetic acid (NAA), without GA(3), were analyzed by polyacrylamide gel electrophoresis. The proteins from the former calli included appreciably higher levels of a 31-kDa basic protein (BP31) (pI=8.8) than proteins from the latter. This 31-kDa protein also accumulated in embryogenic calli derived from non-transformed roots. Somatic embryos from hairy root explants, grown on medium without growth regulators, yielded regenerated plantlets with actively growing root systems and shoots that did not bolt. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • Agrobacterium-mediated transformation of Saintpaulia ionantha Wendl.
    S Kushikawa, Y Hoshino, M Mii
    PLANT SCIENCE, 161, 5, 953, 960, ELSEVIER SCI IRELAND LTD, 2001年10月, [査読有り]
    英語, 研究論文(学術雑誌), Agrobacterium tumefaciens strain LBA4404 that has pTOK233 and other two strains that have pIG121Hm were co-cultivated with suspension cells of Saintpaulia ionantha Wendl. 'Pink Veil'. These two plasmids contain an intron-gusA reporter gene, hygromycin phosphotransferase gene (hpt) and neomycin phosphotransferase II gene (nptII) as selective markers. After 48 h of co-cultivation, the cells inoculated with A. tumefaciens strain LBA4404 (pTOK233) showed the highest GUS activity. After 4 months of subculture on the selection medium supplemented with 50 mg l(-1) hygromycin B, resistant calli appeared and adventitious shoots were regenerated 6 months after the transfer on B5 medium supplemented with 1 mg l(-1) BA. The presence of the gusA gene in the genome DNA of regenerated plants was detected by PCR and genomic Southern blot analysis. Leaf lam ina, another explant used for inoculation, rarely expressed transient GUS activity, and transgenic plants were not obtained. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
  • Isolation of embryo sacs from Dianthus ovules by enzymatic treatments and microdissection
    Y Hoshino, E Nishino, M Mii
    PLANT CELL REPORTS, 19, 5, 443, 447, SPRINGER VERLAG, 2000年04月, [査読有り]
    英語, 研究論文(学術雑誌), Mature ovules of Dianthus (Caryophyllaceae) were histologically observed by clearing and serial sectioning to characterize the cells of the embryo sac. The results show that the mature embryo sac was located deep inside the hemitropous ovule due to thick nucellar tissue at the micropylar region. For the isolation of the embryo sacs, ovules were collected from ovaries of flowers 1 day after anthesis, and treated with an enzyme solution for digesting cell walls on a gyratory shaker. After 12h of enzyme treatment, these ovules were dissected using a glass needle under an inverted microscope to release the embryo sacs. The embryo sacs, characterized by their specific size, were successfully released by these successive treatments. The viability of the embryo sacs was more than 80% as assessed with fluorescein diacetate staining. Fluorescent staining with 4,6-diamidino-2-phenylindole. revealed the nuclei of the egg apparatus in the isolated embryo sacs. The procedure for isolating embryo sacs established in this study will offer a new approach to further in vitro studies on fertilization in Dianthus.
  • Histological examination of callogenesis and adventitious embryogenesis in immature ovary culture of grapevine (Vitis vinifera L.)
    M Nakano, Y Watanabe, Y Hoshino
    JOURNAL OF HORTICULTURAL SCIENCE & BIOTECHNOLOGY, 75, 2, 154, 160, HEADLEY BROTHERS LTD, 2000年03月, [査読有り]
    英語, 研究論文(学術雑誌), Histological examinations of callogenesis and adventitious embryogenesis in immature ovary culture of grapevine (Vitis vinifera L. 'Neo Mat') were carried out during different phases of ontogenetic development. Adventitious embryos and/or embryogenic calli were obtained when immature ovary explants were cultured on a callus induction medium (C medium) for two months followed by transfer of callus-forming explants onto an embryogenesis induction medium (E medium). Microscopic observations of serial sections of the explants revealed that the calli formed on C medium were initiated preferentially from receptacle parenchyma cells. No cell division in the embryo sac was observed and most of them degenerated four weeks after the onset of culture. Two to four weeks after transfer of the explants onto E medium, calli characterized by dense cytoplasm, conspicuous nuclei and thick cell walls were newly formed in the initially-formed, receptacle-derived ones. Proembryos simultaneously developed in the newly formed calli, indicating that they were embryogenic calli. Cell division of embryo sacs was never observed even on E medium, and adventitious embryos and embryogenic calli were hence of somatic origin. Adventitious embryos developed asynchronously and passed through globular, heart, torpedo and cotyledonary stages. These adventitious embryos germinated and developed into plantlets following their transfer onto a plant growth regulator-free medium.
  • Histological examination of callogenesis and adventitious embryogenesis in immature ovary culture of grapevine (Vitis vinifera L.)
    M. Nakano, Y. Watanabe, Y. Hoshino
    Journal of Horticultural Science and Biotechnology, 75, 2, 154, 160, Headley Brothers Ltd, 2000年, [査読有り]
    英語, 研究論文(学術雑誌), Histological examinations of callogenesis and adventitious embryogenesis in immature ovary culture of grapevine (Vitis vinifera L. 'Neo Mat') were carried out during different phases of ontogenetic development. Adventitious embryos and/or embryogenic calli were obtained when immature ovary explants were cultured on a callus induction medium (C medium) for two months followed by transfer of callus-forming explants onto an embryogenesis induction medium (E medium). Microscopic observations of serial sections of the explants revealed that the calli formed on C medium were initiated preferentially from receptacle parenchyma cells. No cell division in the embryo sac was observed and most of them degenerated four weeks after the onset of culture. Two to four weeks after transfer of the explants onto E medium, calli characterized by dense cytoplasm, conspicuous nuclei and thick cell walls were newly formed in the initially-formed, receptacle-derived ones. Proembryos simultaneously developed in the newly formed calli, indicating that they were embryogenic calli. Cell division of embryo sacs was never observed even on E medium, and adventitious embryos and embryogenic calli were hence of somatic origin. Adventitious embryos developed asynchronously and passed through globular, heart, torpedo and cotyledonary stages. These adventitious embryos germinated and developed into plantlets following their transfer onto a plant growth regulator-free medium.
  • Transgenic bialaphos-resistant snapdragon (Antirrhinum majus L.) produced by Agrobacterium rhizogenes transformation
    Y Hoshino, Turkan, I, M Mii
    SCIENTIA HORTICULTURAE, 76, 1-2, 37, 57, ELSEVIER SCIENCE BV, 1998年07月, [査読有り]
    英語, 研究論文(学術雑誌), Transgenic herbicide-resistant snapdragon (Antirrhinum majus L.: Scrophulariaceae) plants were obtained by co-cultivation of leaf explants with Agrobacterium rhizogenes harboring Ri plasmid and pARK5. The T-DNA region of pARK5 contains bar gene encoding phosphinothricin acetyltransferase under the control of the cauliflower mosaic virus 35S promoter, and nptII gene encoding neomycin phosphotransferase II under the control of nopaline synthase promoter. Transformed hairy roots induced from leaf explants inoculated with A. rhizogenes strain A13 (pARK5) could grow on 1 mg l(-1) bialaphos-containing half-strength MS medium, although hairy roots transformed by the wild type strain of A. rhizogenes A13 could not survive at the same concentration of bialaphos. Shoot regeneration from the hairy roots integrating T-DNA of pARK5 was also stimulated at 1 mg l(-1) bialaphos. Regenerated shoots readily developed roots on 2 g l(-1) gellan gum solidified half-strength MS medium. The transgenic nature of the plants was confirmed by opine, PCR and Southern analyses. Regenerated plantlets were successfully established in the growth chamber. Acclimatized plants exhibited altered phenotypes known as Ri syndrome such as reduced apical dominance with highly branched stems and short internodes, and also showed resistance to an applied standard level of commercial herbicides. (C) 1998 Elsevier Science B.V. All rights reserved.
  • Bialaphos stimulates shoot regeneration from hairy roots of snapdragon (Antirrhinum majus L.) transformed by Agrobacterium rhizogenes
    Y Hoshino, M Mii
    PLANT CELL REPORTS, 17, 4, 256, 261, SPRINGER VERLAG, 1998年02月, [査読有り]
    英語, 研究論文(学術雑誌), Hairy roots of snapdragon (Antirrhinum majus L.: Scrophulariaceae) induced by a wild-type strain of Agrobacterium rhizogenes were cultured on media containing various concentrations of a phosphinothricin-based herbicide, bialaphos, or plant growth regulators (PGRs). Adventitious shoot regeneration from hairy roots was observed with a low frequency (10%) on half-strength Murashige and Skoog medium. Addition of alpha-naphthaleneacetic acid in combination with 6-benzylaminopurine, thidiazuron, or zeatin to the medium had no effect on shoot regeneration from hairy roots. Although bialaphos at 0.9 mg l(-1) or more was toxic to hairy roots, it significantly increased the shoot regeneration frequency up to 56% at 0.5 mg l(-1). In contrast, non-transformed roots and leaves regenerated no shoots on media with or without bialaphos. Regenerated shoots detached from host roots readily developed roots on gellan-gum-solidified medium. Regenerated plants were successfully transferred to the greenhouse, but did not produce seed.
  • Highly efficient system of plant regeneration from protoplasts of grapevine (Vitis vinifera L) through somatic embryogenesis by using embryogenic callus culture and activated charcoal
    YM Zhu, Y Hoshino, M Nakano, E Takahashi, M Mii
    PLANT SCIENCE, 123, 1-2, 151, 157, ELSEVIER SCI IRELAND LTD, 1997年03月, [査読有り]
    英語, 研究論文(学術雑誌), A simple protocol is described for high frequency plant regeneration from protoplasts isolated from leaf-derived embryogenic calli of grapevine (Vitis vinifera L. cv. Koshusanjaku). The protoplasts successfully divided to form somatic embryos by culturing in gellan gum disc-method in which protoplasts were embedded in 2 g/l gellan gum-solidified Nitsch's medium containing 2.0 mg/l NAA, 0.5 mg/l BA, 0.09 M sucrose and 0.3 M glucose at a density of 1 x 10(5) protoplasts/ml. For the continuous growth of the colonies without browning, it was essential to add 0.3% (w/v) AC in the liquid reservoir medium from the beginning of the culture. In this culture condition, protoplasts started to divide after IO days of culture and grew into torpedo embryos 4 months after initiation of culture. The torpedo embryos thus obtained germinated normally by transferring onto 2 g/l gellan gum-solidified PGR-free Nitsch's medium containing 30 g/l sucrose. The regenerated plants were successfully transferred to the greenhouse and showed normal morphology. (C) 1997 Elsevier Science Ireland Ltd.
  • Intergeneric somatic hybrid plantlets between Dianthus barbatus and Gypsophila paniculata obtained by electrofusion
    M Nakano, Y Hoshino, M Mii
    THEORETICAL AND APPLIED GENETICS, 92, 2, 170, 172, SPRINGER VERLAG, 1996年02月, [査読有り]
    英語, 研究論文(学術雑誌), Hypocotyl-derived protoplasts of Dianthus barbatus that had been pretreated with iodoacetamide were fused electrically with cell suspension culture-derived protoplasts of Gypsophila paniculata that could divide to form callus but could not regenerate shoots under the culture conditions used in this study, Electrofusion-derived calli which produced shoots were selected as putative somatic hybrids, and plantlets were subsequently regenerated from 2 of these selected calli. These plantlets, which in vitro produced flowers precociously, were identified as intergeneric somatic hybrids by nuclear ribosomal DNA analysis. Normal plants have not been established up to the present.
  • PLANT-REGENERATION FROM CELL SUSPENSION-DERIVED PROTOPLASTS OF SAINTPAULIA-IONANTHA WENDL
    Y HOSHINO, M NAKANO, M MII
    PLANT CELL REPORTS, 14, 6, 341, 344, SPRINGER VERLAG, 1995年03月, [査読有り]
    英語, 研究論文(学術雑誌), Friable calli were induced on leaf segments of Saintpaulia ionantha Wendl. on B5 medium containing 1 mg l(-1) 2,4-D and 2 g l(-1) casein hydrolysate. Cell suspension cultures were readily established from these friable calli and protoplasts could be isolated from the cells with yields of 1-3 x 10(7)/g f. wt.. By culturing in 0.1 % gellan gum-solidified B5 medium supplemented with 1 mg l(-1) 2,4-D and 0.1 M each of sucrose and mannitol at a density of 1 x 10(5)/ml, the protoplasts divided within 6 days and formed macro-colonies after 2 months of culture. Shoot regeneration from protoplast-derived calli was obtained by sequential treatment of the calli with plant growth regulators: initially with 1 mg l(-1) each of NAA and BA for 2 months followed by 0.01 mg l(-1) NAA and 5 mg l(-1) BA for 4 months. Regenerated plants were established after rooting of the shoots on half-strength MS medium, and successfully transferred to the greenhouse. The regenerated plants grew into flowering stage and showed the same phenotype as the parent plant.
  • REGENERATION OF TRANSGENIC PLANTS OF GRAPEVINE (VITIS-VINIFERA L) VIA AGROBACTERIUM-RHIZOGENES-MEDIATED TRANSFORMATION OF EMBRYOGENIC CALLI
    M NAKANO, Y HOSHINO, M MIL
    JOURNAL OF EXPERIMENTAL BOTANY, 45, 274, 649, 656, OXFORD UNIV PRESS UNITED KINGDOM, 1994年05月, [査読有り]
    英語, 研究論文(学術雑誌), Genetically transformed roots and calli were induced from leaf segments of grapevine (Vitis vinifera L. cv. Koshusanjaku) after co-cultivation with wild-type Agrobacterium rhizogenes strains, but plant regeneration from them was not achieved. On the other hand, transgenic grapevine plants were obtained via somatic embryogenesis after co-cultivation of embryogenic calli with an engineered A. rhizogenes strain including both the neomycin phosphotransferase II (NPT II) and the beta-glucuronidase (GUS) genes, followed by selection of secondary embryos for kanamycin resistance. All these plants showed GUS gene expression revealed by histochemical assay. Southern blot analysis revealed the stable integration of the GUS cording region in their genome. Transformants containing Ri T-DNA exhibited various phenotypes: most of them showed a typical Ri-transformed phenotype such as wrinkled leaves, while the others looked normal.
  • ADVENTITIOUS SHOOT REGENERATION FROM CULTURED PETAL EXPLANTS OF CARNATION
    M NAKANO, Y HOSHINO, M MII
    PLANT CELL TISSUE AND ORGAN CULTURE, 36, 1, 15, 19, KLUWER ACADEMIC PUBL, 1994年01月, [査読有り]
    英語, 研究論文(学術雑誌), Adventitious shoot regeneration was compared among leaf, stem and petal explants of carnation (Dianthus caryophyllus L.) cv. Scania on MS medium containing different concentrations of 6-benzyladenine (BA) and alpha-naphthaleneacetic acid (NAA). High frequency regeneration was obtained only from petal explants on the media containing 5 to 10 muM BA with or without 5 muM NAA. Among the cytokinins tested, N-2-chloro-4-pyridyl-N'-phenylurea and N-1,2,3-thiadiazol-5-yl-N'-N'-phenyl-urea were more effective than BA, kinetin, N6-2-isopentenyl adenine and zeatin on regeneration from petal explants. Although, high frequency shoot regeneration was obtained from all petal explants harvested from various developmental stages of buds, a1 significant decrease in regeneration capacity was observed in the explants obtained from fully-opened flowers. High frequency shoot regeneration was also obtained from the petal explants of cvs. Coral, Lena, Nora and White Sim, and an interspecific cultivar Eolo using the method developed in this study.

所属学協会

  • 日本植物学会               
  • Turkish Journal of Botany               
  • 日本植物細胞分子生物学会               
  • 日本園芸学会               
  • 日本育種学会               
  • Japanese Society for Plant Cell and Molecular Biology               
  • Japanese Society for Horticultural Science               
  • Japanese Society of Breeding               

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