Papers - KUNITAKE Hisato
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カンキツ属 4 種とキンカン属 5 種を用いた カンキツ類の遺伝解析に有用な SSR マーカーの選定 Reviewed
安田喜一・戸村大河・佐藤真衣・富永晃好・八幡昌紀・國武久登
東海大学紀要 農学部 41 29 - 37 2022.4
Authorship：Last author Language：Japanese Publishing type：Research paper (scientific journal)
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Mutagenic Effects of Heavy-Ion Beam Irradiation to Plant Genome Invited Reviewed
Hirano Tomonari, Kazama Yusuke, Kunitake Hisato, Abe Tomoko
ＣＹＴＯＬＯＧＩＡ 87 ( 1 ) 3 - 6 2022.3
Language：English Publishing type：Research paper (scientific journal) Publisher：公益財団法人 日本メンデル協会
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Effective Utilization of Vaccinium virgatum Aiton Stems as Functional Materials: Major Constituent Analysis and Bioactivity Evaluation Invited Reviewed
Hisahiro Kai 1, Kazuhiro Sugamoto , Saki Toshima, Yo Goto , Takayuki Nakayama , Kazuhiro Morishita and Hisato Kunitake
Plants 11 ( 568 ) 2022.2
Authorship：Last author Language：English Publishing type：Research paper (scientific journal)
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Inhibition of ATL cell proliferation by polymerized proanthocyanidin from blueberry leaves through JAK proteolysis. Reviewed
Ichikawa T, Sugamoto K, Matsuura Y, Kunitake H, Shimoda K, Morishita K
Cancer science 2022.1
Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1111/cas.15277
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Induction and Characterization of Chromosome-doubled Nagabo-natsuhaze (Vaccinium sieboldii Miq.), a Wild Species Native to Japan, by Colchicine and Oryzalin Treatments of in vitro Shoots Invited Reviewed
Horticultural Research 21 ( 1 ) 1 - 10 2022.1
Authorship：Last author Language：Japanese Publishing type：Research paper (scientific journal)
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Sudo M., Yasuda K., Yahata M., Sato M., Tominaga A., Mukai H., Ma G., Kato M., Kunitake H.
Agronomy 11 ( 12 ) 2021.12
Authorship：Last author Language：English Publishing type：Research paper (scientific journal) Publisher：Agronomy
The morphological characteristics and fruit quality of an autotetraploid plant selected from nucellar seedlings of Satsuma mandarin (Citrus unshiu Marcow.) were investigated. Additionally, in order to evaluate the reproductive potential of male and female gametes of the tetraploid Satsuma mandarin, reciprocal crosses with diploid cultivars were also carried out. The tetraploid had significantly thick and round leaves, as compared to those of the diploid Satsuma mandarin. The sizes of the flowers and pollen grains of the tetraploid were significantly larger than those of the diploid. Pollen fertility of tetraploid was high compared with that of the diploid. The tetraploid produced seedless fruits. The fruit weight of the tetraploid was equal to that of the diploid. Compared to the diploid fruits, the tetraploid fruit had less sugar contents and more organic acid contents. Although the tetraploid fruits showed similar traits to other Citrus tetraploids such as thick and hard peels, the tetraploid had a higher content of carotenoids in the flavedo than the diploid, and the rind color of the tetraploid was much better. In the reciprocal crosses between the tetraploid Satsuma mandarin and diploid cultivars, some seeds were obtained, and triploid progenies were obtained in all cross combinations.
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Yamasaki M., Kiue Y., Fujii K., Sushida M., Yamasaki Y., Sugamoto K., Suzuki Y., Koga Y., Kunitake H., Kai H., Ogawa K., Nishiyama K., Goto Y., Nakayama T.
Plants 10 ( 12 ) 2021.12
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Plants
Blueberry (Vaccinium virgatum Aiton; Kinisato 35 Gou) leaves have recently attracted in-creasing attention as a useful material for the prevention of lifestyle diseases. Here, we examined the effects of the hot water extract of blueberry leaves (BLEx) on lipogenesis and uric acid production in 3T3-L1 adipocytes. The results showed that BLEx suppressed lipid accumulation and the mRNA expression of differentiation markers in 3T3-L1 adipocytes. A fractionation study showed that the highly polymerized proanthocyanidin-rich fraction was responsible for this effect. Upon maturation to adipocytes, 3T3-L1 cells produced uric acid and tumor necrosis factor-α, and hypoxia stimulated the production of uric acid and xanthine oxidoreductase activity. BLEx suppressed the production of uric acid under these conditions. Although BLEx inhibited the enzymatic activity of xanthine oxidase, this activity was observed in several fractions containing catechin, epicatechin, chlorogenic acid, rutin, and low molecular weight proanthocyanidins. Taken together, these results indicate that BLEx contains various compounds with the ability to suppress lipid accumulation and uric acid production in adipocytes.
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Fruit Qualities of Interspecific Hybrid and First Backcross Generations between Red Raspberry and Rubus parvifolius Reviewed
Saki Toshima, Marika Fujii, Momoko Hidaka, Soya Nakagawa, Tomonari Hirano, Hisato Kunitake
J. Amer. Soc. Hort. Sci. 146 ( 6 ) 445 - 451 2021.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Yasuda K., Yahata M., Sato M., Sudo M., Tominaga A., Kunitake H.
Agronomy 11 ( 10 ) 2 - 16 2021.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Agronomy
We previously obtained two intergeneric hybrids with different ploidies, i.e., aneuploid (2n = 28) and eutriploid, from diploid−diploid crosses between ‘Kiyomi’ tangor (Citrus unshiu Marcow. × C. sinensis (L.) Osbeck) and Meiwa kumquat (Fortunella crassifolia Swingle) as novel breeding materials for a seedless kumquat. In this study, we attempted to clarify the construction of the parental genomes of these hybrids by SSR genotyping and genomic in situ hybridization (GISH)−chromomycin A3 (CMA) analysis. SSR genotyping in NSX43 (LG5) and CiBE2227 (LG8) loci revealed that both hybrids inherited one allele from ‘Kiyomi’ tangor and two heterozygous alleles from Meiwa kumquat. The GISH analysis failed due to the high genomic homology between Citrus and Fortunella. At the same time, the CMA karyotype compositions of the two intergeneric hybrids (H15-701: 2A + 1B + 3C + 13D + 7E + 1F + 1Dst; H15-702: 3A + 1B + 2C + 15D + 4E +1F + 1Dst) and both parents (‘Kiyomi’ tangor: 1A + 2B + 2C + 6D + 7E; Meiwa kumquat: 2A + 2C + 12D + 1F + 1Dst) were completely revealed. We identified the parental genome construction and polyploidiza-tion processes in both intergeneric hybrids on the basis of these SSR genotypes and CMA karyotype compositions according to the following theory: the SSR genotypes and chromosome compositions were the same as those of the somatic chromosome and two-fold after the first division (even num-ber) in unreduced gametes caused by first-division restitution (FDR) and second-division restitution (SDR), respectively. Consequently, we determined that both intergeneric hybrids may have had two genomes derived from the 2n male unreduced gamete as a result of the FDR of the Meiwa kumquat. In addition, most horticultural traits of the leaves, flowers, and fruits of both hybrids showed intermediate traits of the parents, but the fruit sizes and flowering habits were more like those of the two inherited genomes of Meiwa kumquat.
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宮崎大学医学部附属病院臨床研究支援センター食品臨床試験部門の取り組み Reviewed
有村保次、永濱清子、黒木光渉、國武久登、片岡寛章（他6名）
宮崎県医師会医学会誌 45 ( 2 ) 222 - 230 2021.9
Language：Japanese Publishing type：Research paper (scientific journal)
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Nhat V.Q., Kazama Y., Ishii K., Ohbu S., Kunitake H., Abe T., Hirano T.
Plants 10 ( 9 ) 2021.9
Language：English Publishing type：Research paper (scientific journal) Publisher：Plants
Two growth processes, cell proliferation and expansion, determine plant species-specific organ sizes. A large flower mutant in Arabidopsis thaliana, ohbana1 (ohb1), was isolated from a mutant library. In the ohb1 flowers, post-mitotic cell expansion and endoreduplication of nuclear DNA were promoted. The whole-genome resequencing and genetic analysis results showed that the loss of function in MEDIATOR16 (MED16), a mediator complex subunit, was responsible for the large flower phenotypes exhibited by ohb1. A phenotypic analysis of the mutant alleles in MED16 and the double mutants created by crossing ohb1 with representative large flower mutants revealed that MED16 and MED25 share part of the negative petal size regulatory pathways. Furthermore, the double mutant analyses suggested that there were genetically independent pathways leading to cell size restrictions in the floral organs which were not related to the MED complex. Several double mutants also formed larger and heavier seeds than the wild type and single mutant plants, which indicated that MED16 was involved in seed size regulation. This study has revealed part of the size-regulatory network in flowers and seeds through analysis of the ohb1 mutant, and that the size-regulation pathways are partially different between floral organs and seeds.
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Saki Toshima, Tomonari Hirano, Hisato Kunitake
Scientia Horticulturae 285 2021.7
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
The breeding of raspberries (Rubus idaeus L.) and blackberries (R. fruticosus Agg.) has shifted in focus toward traits associated with the sensory qualities and potential health benefits of the fruit, and Rubus wild species have attracted a great deal of attention as new sources of genetic diversity. In this study, the anthocyanin and polyphenol contents and compositions, and three types of antioxidant capacities were compared in mature fruits including raspberries, blackberries, and Japanese wild Rubus species. ‘Blackcap’ black raspberry showed the significantly highest values of total anthocyanin content and total polyphenol content among 12 Rubus species or cultivars. Cyanidin-3-xylosyl-rutinoside was contained only in black raspberries, and was characteristic anthocyanin in Rubus species. R. croceacanthus H. Lév., a Japanese wild species that is adapted to warm regions, had a deep red fruit color, and had the highest total anthocyanin and total polyphenol contents among the four Japanese wild species examined. As for the antioxidant capacities, the highest values were observed in ‘Blackcap’ among 12 Rubus species. Furthermore, a high positive correlation was shown between total anthocyanin or total polyphenol content and antioxidant capacities. In particular, black raspberries and blackberries contained high concentrations of rutin. Thus, rutin might contribute to the high antioxidant capacities in these cultivars.
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Nakagawa S., Ohmura R., Toshima S., Park H., Narasako Y., Hirano T., Otani M., Kunitake H.
Scientia Horticulturae 284 2021.6
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae
Anthocyanins and polyphenols in the sweetpotato (Ipomoea batatas L.) have attracted attention due to their multiple physiological functions. Therefore, sweetpotato breeders have focused on increasing the anthocyanin and polyphenol contents. In this study, we investigated the diversity of anthocyanins, polyphenols, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activities in sweetpotato genotypes. We also assessed how the anthocyanin and polyphenol contents and compositions were related to DPPH radical-scavenging activities during tuber growth in several cultivars of sweetpotatoes. In six purple-fleshed cultivars, the percentage of acylated anthocyanins that were evaluated was high (92.0–98.5 %), whereas in ‘Fukumurasaki’ this percentage was particularly low (63.6 %). Moreover, the percentages of anthocyanins that were acylated in the purple-fleshed cultivars ‘Tanegashimamurasaki’, ‘Purple Sweet Lord’, and ‘Akemurasaki’ had already reached high levels in the early stage of tuber development. On the other hand, these three cultivars with high anthocyanin contents had high DPPH radical-scavenging activities, and the activities during tuber growth showed a high correlation with total anthocyanin content. Elucidating the changes in acylated anthocyanin content and in antioxidant activities during tuber growth can help promote high-quality sweetpotato cultivation and breeding.
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Polyphenol contents, antioxidant activities, and anti-cancer cell proliferation properties at each stage of fruit development in intersectional hybrids between highbush blueberry and shashanbo (Vaccinium bracteatum Thunb.) Invited Reviewed
Toyama Yuki, | Toshima, Saki, Hirano, Tomonari, Yamasaki, Masao, Kunitake, Hisato
Journal of Berry Research 1 - 16 2021.6
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.3233/JBR-210713
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Comparison of fruits quality and antioxidant activity between diploid and tetraploid in Satsuma mandarin Reviewed
Mai SATO, Kiichi YASUDA, Kohei KIKUCHI, Taiga TOMURA, Masaki YAHATA and Hisato KUNITAKE
Proceedings of the School of Agriculture, Tokai University 40 1 - 8 2021.3
Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
To clarify the time-dependent change of fruit quality and health functional components in the tetraploid Satsuma mandarin
(Citrus unshiu Marcow.), the fruits were measured the coloring fruit skin, sugar and organic acid contents, total polyphenol
content and antioxidant activities by ABTS cation radical scavenging activity. As a result, the tetraploid fruit skin indicated the
deeper and redder color than that of the diploid at harvesting stage. There was no significant difference in the sugar contents
between the diploid and tetraploid. The organic acid content generally continued to be flat in the tetraploid pulp (juice sac),
whereas it was normally decreased in the diploid pulp with maturation of fruits. For the last period, this value of the tetraploid
was significantly higher than the diploid. Furthermore, the antioxidant activity the flavedo and/or albedo in the tetraploid was
significantly higher than the diploid during some investigation periods, although there was no significant difference between
the two in the total polyphenol content, expect for some investigation periods. Thus, it was considered that the tetraploid
Satsuma mandarin was difficult to use as a fresh fruit, but it was useful as a processed food such as marmalade, dried fruit and
juice whose raw material was pericarp. -
Relationship between morphology and weight of seeds and ploidy level of seedlings obtained by cross between diploid pummelo 'Banpeiyu' and triploid gragpefruit-type citrus 'Orobranco'. Reviewed
20 ( 1 ) 29 - 37 2021.3
Language：Japanese Publishing type：Research paper (scientific journal)
DOI： 10.2503/hrj.20.29
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Mik iKawano, Masaki Yahata, Tokurou Shimizu, Chitose Honsho, Tomonari Hirano, Hisato Kunitak
Scientia Horticulturae 277 2021.2
Language：English Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae
The genetic analysis of five haploid plants (BX1-BX5) of ‘Banpeiyu’ pummelo (BP), [Citrus maxima (Burm.) Merr.] using 32 simple sequence repeat (SSR) markers carried out for the purpose of confirming their gynogenetic origin and genetic differences. As the results, all the haploid plants possessed only either one of the BP alleles, confirming that they are gynogenetic origin. We also confirmed that BX1 and its doubled haploid (BX1-DH) have exactly the same SSR alleles. Interestingly, all the five haploids had common alleles on the linkage group (LG) 6 in the case of three markers. Then, we performed self-pollination of DH plants and reciprocal crosses between DH and BP, which has strong self-incompatibility. The seeds obtained by these crosses germinated in vitro or ex vitro normally and developed into diploid seedlings, except for one tetraploid seedling. Genetic analysis of these seedlings with 32 SSR markers revealed that segregation ratios of the 32 loci in the 30 BP inbred, 30 BP x DH and 30 DH x BP progenies mostly fitted to the expected ratios. However, significantly distorted segregation ratios were found in one, five and three SSR markers for BP inbred, BP x DH and DH x BP progenies, respectively. All of the 30 DH inbred progenies showed exactly the same SSR alleles as the original DH. This is the first example in which self and back cross progenies of DH in fruit crops were genetically analyzed using SSR markers.
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DNA Damage Response of Cyrtanthus mackenii Male Gametes Following Argon Ion Beam Irradiation Reviewed
Hirano T., Matsuyama Y., Hanada A., Hayashi Y., Abe T., Kunitake H.
Cytologia 86 ( 4 ) 311 - 315 2021
Language：English Publishing type：Research paper (scientific journal) Publisher：Cytologia
Heavy-ion beams have been applied as effective mutagens to various plant materials. Pollen has been used as material for mutant induction and genetic analysis. However, our knowledge of the DNA damage response of plant male gametes remains limited. In the present study, we irradiated Cyrtanthus mackenii pollen with an argon ion beam, which induced complex DNA damage, and investigated the DNA damage response of male gametes during pollen tube growth. Male gametes derived from the irradiated pollen grains were isolated from pollen tubes after 12 and 24 h of culture and subjected to cell cycle analysis. After 12 h of culture, the irradiated generative cells were frequently arrested at metaphase during pollen mitosis II (PMII), and the proportion of metaphase cells increased with increasing absorbed dose. These results suggest that the genomic lesions induced by the argon ion beam caused spindle assembly checkpoint (SAC)-dependent arrest. After 24 h of culture, the irradiated male gametes completed PMII, albeit forming sperm cells with abnormalities in chromosome separation, and chromosomal bridges were often formed between these cells. Moreover, phosphorylated H2AX foci, an indicator of DNA double-strand breaks, were detected in the irradiated male gametes after 24 h of culture, regardless of passing through the SAC. Taken together, these results indicate that male gametes activate functions to cope with radiation-induced complex DNA damage during pollen tube growth.
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Phylogeny and trait variation of Japanese Rubus subgenus Ideaobatus Reviewed
Okada A., Kikuchi S., Hoshino Y., Kunitake H., Mimura M.
Scientia Horticulturae 264 2020.3
Language：English Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae
© 2019 Elsevier B.V. Rubus L. subgenus Idaeobatus is one of the largest groups comprising mostly diploid species within the genus. Idaeobatus species is distributed throughout Japan, showing a wide range of trait variation and climatic distribution. We constructed phylogenetic trees of Idaeobatus that are naturally distributed in Japan. The Bayesian and ML phylogenetic tree found that this subgenus was polyphyletic with at least three genetic groups. The phylogenetic tree of Japanese Idaeobatus also reflects the trait variations for flower colors, fruit colors, and leaf shape (simple vs compound). The species that occurs in subtropical climates was found to be closely related to their sister temperate species in Japanese Idaeobatus. Crossability generally reflected the phylogenetic distances between species. Trait diversity and its phylogenetic relationship among Japanese Idaeobatus can provide additional insights for future breeding and ecological and evolutionary studies.
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Shoji K., Yamasaki M., Kunitake H.
Journal of Oleo Science 69 ( 2 ) 143 - 151 2020.2
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Oleo Science
©2020 by Japan Oil Chemists’ Society. Prevention of postprandial hypertriglyceridemia is an important consideration for reducing the risk of developing cardiovascular disease. While blueberry fruits have been reported to ameliorate lipid metabolism in humans, there are only few research reports on the effects of blueberry leaves (BL). Here, we investigated the efficacy of BL on postprandial hyperlipidemia in subjects with high fasting triacylglycerol (TG) concentrations. Randomized, double-blind, cross-over design study was conducted. The subjects consumed a BL containing beverage or a placebo beverage before a fat-enriched test meal. Blood samples were collected prior to and 1, 2, 3, 4, and 5 hours after consuming the test beverage. The postprandial serum TG and remnant-like particle cholesterol (RLP-C) concentrations were significantly lower in the BL beverage compared with those in the placebo beverage. Additionally, BL was more effective in subjects with high fasting ghrelin with gastric emptying function. In current study, fasting ghrelin correlated with the increase in postprandial serum TG, suggesting that BL ameliorates hypertriglyceridemia through delayed gastric emptying. In conclusion, this pilot study suggests that BL may be useful as an early dietary therapy for treating postprandial hyperlipidemia.
DOI： 10.5650/jos.ess19201