論文 - 國武 久登
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宮崎大学における全学的な設備共同利用体制の構築:～学内既存システムとリンクした設備共通管理システムの開発～
境 健太郎, 今井 正人, 出水 信雄, 神力 はるな, 國武 久登, 三澤 尚明, 水光 正仁
研究 技術 計画 35 ( 1 ) 23 - 31 2020年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：研究・イノベーション学会
Recently, many universities have been working to promote the sharing of research facilities and equipment in Japan. University of Miyazaki is also developing a sharing service system of those through two projects supported by the Ministry of Education, Culture, Sports, Science and Technology. In addition, we have established the Miyazaki Facility Network, in which public research institutions in Miyazaki Prefecture participate, and are working to create an environment that can provide mutually advanced analysis technologies that contribute to regional revitalization.In this paper, we introduce a common facilities management system linked on-line to the existing financial accounting and the researcher database developed by University of Miyazaki, and describe the relationship with the administration policy of our university and examples of utilization in Miyazaki prefecture.
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Nukaya T., Sudo M., Yahata M., Ohta T., Tominaga A., Mukai H., Yasuda K., Kunitake H.
Agronomy 9 ( 9 ) 2019年9月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Agronomy
© 2019 by the authors. A ploidy chimera of the Meiwa kumquat (Fortunella crassifolia Swingle), which had been induced by treating the nucellar embryos with colchicine, and had diploid (2n = 2x = 18) and tetraploid (2n = 4x = 36) cells, was examined for its ploidy level, morphological characteristics, and sizes of its cells in its leaves, flowers, and fruits to reveal the ploidy level of each histogenic layer. Furthermore, the chimera was crossed with the diploid kumquat to evaluate the ploidy level of its reproductive organs. The morphological characteristics and the sizes of the cells in the leaves, flowers, and fruits of the chimera were similar to those of the tetraploid Meiwa kumquat and the ploidy periclinal chimera known as "Yubeni," with diploids in the histogenic layer I (L1) and tetraploids in the histogenic layer II (L2) and III (L3). However, the epidermis derived from the L1 of the chimera showed the same result as the diploid Meiwa kumquat in all organs and cells. The sexual organs derived from the L2 of the chimera were significantly larger than those of the diploid. Moreover, the ploidy level of the seedlings obtained from the chimera was mostly tetraploid. In the midrib derived from the L3, the chimera displayed the fluorescence intensity of a tetraploid by flow cytometric analysis and had the same size of the cells as the tetraploid and the Yubeni. According to these results, the chimera is thought to be a ploidy periclinal chimera with diploid cells in the outermost layer (L1) and tetraploid cells in the inner layers (L2 and L3) of the shoot apical meristem. The chimera had desirable fruit traits for a kumquat such as a thick pericarp, a high sugar content, and a small number of developed seeds. Furthermore, triploid progenies were obtained from reciprocal crosses between the chimera and diploid kumquat.
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Blueberry Leaf Polyphenols Prevent Body Fat Accumulation in Mice Fed High-fat, High-sucrose Diet. 査読あり
Fujii K, Ota Y, Nishiyama K, Kunitake H, Yamasaki Y, Tari H, Araki K, Arakawa T, Yamasaki M
Journal of oleo science 2019年4月
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Subchronic toxicity evaluation of leaves from rabbiteye blueberry (Vaccinium virgatum Aiton) in rats 査読あり
Tanaka , Daigo Yokoyama , Yasushi Matsuura , Masahiko Nozaki , Naoya Hirozawa ,Naoya Hirozawa ,Hisato Kunitake , Masanobu Sakono , Hiroyuki Sakakibara
Toxicology Reports 6 272 - 278 2019年3月
記述言語：英語 掲載種別：研究論文（学術雑誌）
Blueberry leaf may contain multiple compounds with beneficial effects. We conducted a 90-day toxicity study in rats to evaluate the safety of consuming the leaves of rabbiteye blueberry (Vaccinium virgatum Aiton; RB species). Powdered leaves were administered daily by oral gavage at doses of 500, 1000, and 2500 mg/kg body weight to male and female Sprague-Dawley rats for 90 days. Treatment did not result in death or changes in the behavior and external appearance of the animals. No alterations were observed in hematological and serum chemical parameters, urinalysis, food consumption, body weight gain, or absolute and relative organ weights at the end of the treatment period, with the exception of some leukocyte percentages in male rats treated with 500 and 1000 mg/kg blueberry leaf powder. The findings indicate that rabbiteye blueberry leaf is safe for consumption and should be investigated as a candidate functional food.
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Nukaya T., Sudo M., Yahata M., Nakajo Y., Ohta T., Yasuda K., Tominaga A., Mukai H., Kunitake H.
Scientia Horticulturae 245 ( 9 ) 210 - 217 2019年2月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scientia Horticulturae
© 2018 Elsevier B.V. Three Fortunella (kumquat) species, the Meiwa kumquat (F. crassifolia Swingle), the Round kumquat [F. japonica (Thunb.) Swingle] and the Changshou kumquat (F. obovata hort. ex Tanaka), showing tetraploid, which had been induced by treating the seeds with colchicine, were examined for origin and horticultural characteristics (e. g. morphology of leaves, flowers, pollen and fruits). Additionally, these tetraploid kumquats were crossed to produce triploid kumquats. All of the tetraploids were confirmed to be derived from the nucellar embryo of each original kumquat by examining their chromosomal composition with chromomycin A3 staining. All of the tetraploid kumquats had the typical morphological characteristics of tetraploid Citrus plants, such as round and thick leaves, and large flowers and pollen grains. On the other hand, the fruits of these tetraploid plants showed desirable traits for kumquats such as thicker pericarp and higher soluble solids content. Furthermore, when these tetraploids were crossed with some diploid cultivars, triploid progenies were obtained from almost all of the cross combinations.
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Kai H., Uesawa Y., Kunitake H., Morishita K., Okada Y., Matsuno K.
Planta Medica 85 ( 1 ) 81 - 87 2019年1月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Planta Medica
© Georg Thieme Verlag KG Stuttgart. New York. Metabolic profiling is often used to identify possible correlations between a compound's metabolic profile and biological activity. Direct-injection electron ionization-mass spectrometry fingerprinting is useful for characterizing biological materials. We demonstrate the utility of direct-injection electron ionization-mass spectrometry for metabolic profiling using 100 different extracts of leaves from 20 blueberry cultivars collected at 5 time points from April to December 2008. A qualitative direct-injection electron ionization-mass spectrometry method was used to profile the major and/or minor constituents in the blueberry leaf extracts. Blueberry leaf extracts could be distinguished by principal component analysis based on the absolute intensity of characteristic fragment ions. Twenty cultivars were categorized into four species, and the most appropriate discriminative marker m/z value for identifying each cultivar was selected statistically. Correlated m/z values indicating the collection month were determined in the same analysis, and air temperature variance factors were extracted from score plots by principal component analysis. We previously reported that blueberry extracts inhibit the proliferation of adult T-cell leukemia cells. Leaves of Vaccinium virgatum collected in December of 2008 exhibited significantly greater inhibition of adult T-cell leukemia cell proliferation than other species. Highly bioactive cultivars or species were identified by direct-injection electron ionization-mass spectrometry metabolomics analysis of blueberry leaf extracts. The components extracted based on our direct-injection electron ionization-mass spectrometry analyses could be used to construct a model to predict anti-adult T-cell leukemia bioactivity. This is the first study to report a relationship between seasonal variation and bioactivity of natural products using a direct-injection electron ionization-mass spectrometry metabolomics method.
DOI： 10.1055/a-0725-8295
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Erratum: Direct-Injection Electron Ionization-Mass Spectrometry Metabolomics Method for Analyzing Blueberry Leaf Metabolites That Inhibit Adult T-cell Leukemia Proliferation.
Kai H, Uesawa Y, Kunitake H, Morishita K, Okada Y, Matsuno K
Planta medica 85 ( 1 ) e3 2019年1月
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Yasushi Matsuura, Hiroyuki Sakakibara, Maho Kawaguchi, Emi Murayama, Daigo Yokoyama, Chizuko Yukizaki, Hisato Kunitake, Masanobu Sakono
Functional Foods in Health and Disease 8 ( 9 ) 447 - 461 2018年9月
記述言語：英語 掲載種別：研究論文（学術雑誌）
Background: Blueberry stems, a by-product of blueberry leaf tea production, are typically discarded. We evaluated the effects of hot-water extracts of rabbiteye blueberry (Vaccinium virgatum Aiton; RB species) leaves and stems on hepatic lipid levels in rats consuming a high-sucrose diet.
Methods: Male Sprague-Dawley rats were divided into groups that received a control high-sucrose diet alone or supplementation with 2.0% blueberry leaf extract or 0.5% or 2.0% blueberry stem extract. Blood and hepatic lipid levels, hepatic lipogenic enzyme activity, and hepatic quercetin metabolites were evaluated after 28 days of ad libitum consumption.
Results: Supplementation with the extracts did not affect body weight gain, food intake, liver and white adipose tissue weights, or serum lipid levels. Hepatic triglyceride and total cholesterol levels were reduced in the groups that received 2.0% supplementation of either extract. Hepatic malic enzyme activity was also reduced in those groups. Quercetin and its glycosides, the major polyphenols identified in the extracts, accumulated in the liver as quercetin aglycone and quercetin metabolites.
Conclusion: We demonstrated how daily consumption of blueberry leaf and stem extracts can decrease hepatic lipid levels, potentially downregulating malic enzyme activity. These effects were intensive in leaf extracts. The active compounds existed in both extracts may be quercetin and its glycosides. Therefore, blueberry stems and leaves may be an attractive candidate novel functional food. -
YAHATA Masaki, KUNITAKE Hisato, KOMATSU Haruki
Japan Agricultural Research Quarterly: JARQ 51 ( 4 ) 293 - 298 2017年10月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：国立研究開発法人 国際農林水産業研究センター
To obtain basic information about haploid plants in <i>Citrus</i>, the morphological characteristics and reproductive potential of a haploid plant obtained from the 'Banpeiyu' pummelo [<i>C. maxima</i> (Burm.) Merr.] were examined. The leaves, flowers and fruits of the haploid were significantly smaller than those of the 'Banpeiyu' pummelo. Although the haploid showed remarkably reduced pollen fertility compared with the 'Banpeiyu' pummelo, it produced slightly fertile pollen grains. In crosses with diploid cultivars, when the haploid was the seed parent, there was no seed. But when diploid cultivars were pollinated with pollen of the haploid, diploid progenies were obtained. During female gamete formation, no embryo-sac mother cell was formed in the ovules, and consequently no normal embryo sac was observed at anthesis, owing to female sterility caused by the lack of embryo-sac mother cell (EMC) formation in the haploid. During male gamete formation, however, univalents were not distributed to opposite poles at anaphase I in some dividing cells; instead, they remained near the equatorial plate. The nine univalents then performed mitosis, and the univalents were equally distributed to opposite poles. And at the tetrad stage, dyads were also produced in the haploid at a ratio of 24.7%. These results suggest that fertile pollen grains in the haploid would be dyad derivation, produced by abnormalities in the first meiotic division such as first division restitution (FDR).
DOI： 10.6090/jarq.51.293
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ラズベリー‘インディアンサマー’と在来野生種ナワシロイチゴとの種間雑種の生育特性と果実品質 査読あり
登島早紀・岡本（中村）理恵・阿部健一・坂嵜 潮・小松春喜・國武久登
園芸学研究 16 ( 3 ) 345 - 352 2017年8月
記述言語：日本語 掲載種別：研究論文（学術雑誌）
温暖な日本の気候に適し，健康機能性を有するラズベ
リー品種の作出のため，環境適応性の高い在来野生種ナワ
シロイチゴ（Rubus parvifolius L.）とラズベリー‘インディ
アンサマー’（R. idaeus L.）の種間交雑を行った．葉，花お
よび果実の形態的調査において，‘07RUBIXP01’は両親の
中間的な値を示し，RAPD 分析によるDNA 評価でも雑種
性が確認された．‘07RUBIXP01’は小さな刺が見られるも
のの，暖地環境下でも旺盛な生育を示し，両親に比べ果実
重が有意に高いことが確認された．総ポリフェノール，
アントシアニンおよびエラジタンニン含量測定において，
‘07RUBIXP01’は両親の中間的な値を示した．特にエラジ
タンニン含量において‘インディアンサマー’と比べ約4
倍高い値を示した．さらに，糖と有機酸含量の測定では，
両親に比べ糖酸比が高く，食味に優れていた．2012 年に
‘07RUBIXP01’として品種登録し（農林水産省品種登録第
21801 号），今後日本の温暖地域において有望なラズベリー
品種として期待できる． -
Dhungana Sabitri Adhikari, Kunitake Hisato, Niino Takao, Yamamoto Shin-ichi, Fukui Kuniaki, Tanaka Daisuke, Maki Shinya, Matsumoto Toshikazu
Ｐｌａｎｔ Ｂｉｏｔｅｃｈｎｏｌｏｇｙ 34 ( 1 ) 1 - 5 2017年3月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：日本植物細胞分子生物学会
Cryopreservation is an important tool for long-term storage of plant germplasm that is currently used for plant germplasm storage at many institutes worldwide. Recently, novel cryogenic procedures (V and D cryo-plate methods) have been developed. In this study, the most suitable conditions for preserving blueberry shoot tips derived from in vitro and current shoots using the D cryo-plate method were investigated. The D cryo-plate method has advantages such as higher regrowth after cryopreservation and a more user-friendly process compared with conventional cryogenic methods. The optimum duration of desiccation for regrowth of shoot tips from each shoot type was 1 h. To induce dehydration tolerance for the shoot tips, the effects of two cryoprotection treatments (sucrose preculture and loading solution [LS] treatment) on shoot regrowth after cryopreservation were investigated. The combined effect of both treatments significantly increased percentage regrowth (approximately 90%). No regrowth of shoot tips was attained without the two treatments. Thus, preculture and LS treatment were effective to induce dehydration tolerance for cryopreservation of blueberry shoot tips. The optimized conditions for blueberry shoot tips using the D cryo-plate technique were: preculture with 0.3 M sucrose for 1 day, LS treatment (2 M glycerol +0.4–1.0 M sucrose) for 30 min, and air dehydration for 1 h. This optimized procedure was applied to additional blueberry cultivars shoot tips derived from in vitro shoots (regrowth 46.7–100%) and current shoots (regrowth 17.2–62.7%). Furthermore, in vitro shoot tips were suitable material for the D cryo-plate method in blueberry.
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Effect of temperature on self-incompatibility in Citrus pistil and mature pollen culture systems
Kawano S., Li Y., Yahata M., Kunitake H.
Acta Horticulturae 1135 117 - 122 2016年7月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Acta Horticulturae
© 2016, International Society for Horticultural Science. All rights reserved. Temperature has a clear effect on pollen germination and pollen tube growth. Recently, researchers have reported a relationship between self-incompatibility (SI) and temperature. However, the detailed behavior and mechanisms of this relationship have not been completely clarified in Citrus. In the present study, to reveal the relationship between SI and temperature in Citrus cultivars, we investigated pollen germination and pollen tube behavior using in vitro pistil and mature pollen systems under controlled environmental chamber conditions of 25, 30 and 35°C. We first evaluated pollen tube growth for one week in in vitro detached pistils of the SI cultivar 'Hyuganatsu' (Citrus tamurana hort. Ex Tanaka) after self- or cross-pollination. 'Hassaku' (C. hassaku) and 'Banpeiyu' pummelo [C. maxima (Burm.) Merr.] were used as compatible cultivars. Pollen tube growth was observed one week after pollination using aniline blue staining and a fluorescent microscope. Results indicated incompatible pollen tubes stopped in the upper part of styles at 25°C, whereas they reached the base of the styles at 30°C, similar to the behavior of compatible pollen tubes at 25°C. To investigate improvement of the SI-like responses of pollen tubes under different temperature conditions, we examined the inhibition of pollen tube growth and morphological changes of tube tips using the mature pollen culture of 'Hyuganatsu' (Abe et al., in press). The pollen was treated with stylar crude extract protein of 'Hyuganatsu' to simulate the SI-like response. Pollen tubes with the SI treatment showed tip inflation, content leakage and abnormal tip twisting at 25°C, and tube growth was inhibited. However, at 30°C, these SI-like responses were suppressed. These results suggested that temperature might be a trigger for avoiding SI reaction in Citrus.
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Li Y., Fuse T., Kawano S., Kunitake H.
Acta Horticulturae 1135 109 - 116 2016年7月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Acta Horticulturae
© 2016, International Society for Horticultural Science. All rights reserved. Self-incompatibility (SI) in Citrus strongly prevents self-pollination and promotes outcrossing. Even though it has been confirmed that the S-allele is a key gene in controlling the recognition between pollen and pistil in Rosaceae, Solanaceae and Papaveraceae, this has not been cloned in Citrus. A citrus mature pollen culture system was established in our lab in order to culture 'Banpeiyu' and 'Hyuganatsu' pollen in vitro. Stylar crude protein extracts were used to treat citrus pollen in order to induce the SI-like response in the pollen tube. We observed the alteration of the pollen tube shape, including tip twisting and content leakage. Programmed cell death (PCD), as a normal cellular process that eliminates unwanted cells, was reported in previous studies of incompatible pollen tubes, together with an increase in hydrogen peroxide (H 2 O 2 ), depolymerization of actin skeleton, the collapse of mitochondria, and leakage of cytochrome c. In our study, we investigated the cytosolic calcium ion concentration ([Ca 2+ ] cyt ) changes and identified alterations in the actin cytoskeleton in the pollen tube after exposure to the incompatible treatment. The [Ca 2+ ] cyt content increased after incompatible treatment, but there was no obvious difference between the compatible-treatment group and the control. Punctate foci of actin filaments were found in the incompatible-treated pollen tubes; the rates of punctate foci were significantly higher than in the compatible-treated pollen tubes in both 'Banpeiyu' and 'Hyuganatsu'. The results suggested that these alterations in the pollen tubes were directly induced by the SI-like response, and might cause pollen tube PCD.
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Phylogeny and Classification of Kumquats (Fortunella spp.) Inferred from CMA Karyotype Composition 査読あり
Kiichi Yasuda, Masaki Yahata and Hisato Kunitake
The Horticulture Journal 85 ( 2 ) 115 - 121 2016年3月
記述言語：英語 掲載種別：研究論文（学術雑誌）
DOI： 10.2503/hortj.MI-078
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Assessment of genetic relationships among cultivated and wild Rubus accessions using AFLP markers
Miyashita T., Kunitake H., Yotsukura N., Hoshino Y., Hoshino Y.
Scientia Horticulturae 193 165 - 173 2015年9月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scientia Horticulturae
© 2015 Elsevier B.V.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.
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Li, Y., Abe, A., Fuse, T., Hirano, T., Hoshino, Y., Kunitake, H.
Journal of the American Society for Horticultural Science 140 ( 4 ) 339 - 345 2015年8月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Li Y., Abe A., Fuse T., Hirano T., Hoshino Y., Kunitake H.
Journal of the American Society for Horticultural Science 140 ( 4 ) 339 - 345 2015年7月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of the American Society for Horticultural Science
© 2015, American Society for Horticultural Science. All rights reserved. Self-incompatibility (SI) is an important mechanism in higher plants that promotes outcrossing and prevents self-fertilization. ‘Banpeiyu’ (Citrus maxima) and ‘Hyuganatsu’ (Citrus tamurana), two of the Citrus cultivars distributed in Kyusyu, Japan, show gametophytic SI. In this study, we used the Citrus mature pollen culture system and stylar crude protein extracts to simulate compatible (C) and SI responses in ‘Banpeiyu’ pollen tubes. We analyzed the protein changes in pollen tubes with the C- and SI-like treatments by nano-liquid chromatography-mass spectrometry (nano-LC-MS); 14 and 27 proteins were identified in C- and SI-like treatments, respectively. We picked up some candidate genes that were particularly prevalent in SI-like treatment and analyzed their expression level changes during C- and SI-like treatments in ‘Banpeiyu’ and ‘Hyuganatsu’ pollen tubes. The expression levels of copper/zinc superoxide dismutase (Cu/Zn SOD), manganese SOD (Mn SOD), catalase (CAT), and cysteine protease (CYP) increased after SI-like treatment. We used a fluorescent probe to visualize reactive oxygen species (ROS) level changes in ‘Banpeiyu’ and ‘Hyuganatsu’ pollen tubes after C- and SI-like treatments and found that 2-hour SI-like treatment induced ROS levels to increase in the pollen tubes of both cultivars. These results suggest that an ROS increase could be one of the key phenomena in the SI response of Citrus and that gene expression changes were responses to ROS generation.
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Li Yiran, Uchida Aska, Abe Akiha, Yamamoto Akihiro, Hirano Tomonari Kunitake Hisato
Journal of American Society of Horticultural Sciences 140 ( 2 ) 183 - 190 2015年2月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Analysis of expressed proteins in the pollen tube occurring self-incompatible response using mass or single-culture system of mature pollen in Citrus 査読あり
Abe A., Uchida A., Hoshino Y., Sakakibara Y., Suiko M., Kunitake H.
Acta Horticulturae 1267 - 1274 2015年2月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Yahata Masaki, Nukaya Tsunaki, Sudo Miki, Ohta Tomohiro, Yasuda Kiichi, Inagaki Hidehiro, Mukai Hiroo, Harada HIsashi, Takagi Toshihiro, Komatsu Haruki, Kunitake Hisato
The Horticulture Journal 84 ( 1 ) 30 - 36 2015年2月
記述言語：英語 掲載種別：研究論文（学術雑誌）
DOI： 10.2503/hortj.MI-005