Details of a Researcher - KUNITAKE Hisato

Seasonal Variation and Mean Degree of Polymerization of Proanthocyanidin in Leaves and Branches of Rabbiteye Blueberry (Vaccinium virgatum Aiton) Reviewed

Yasuko Koga, Yuno Setoguchi, Kazuhiro Sugamoto, Yo Goto, Tomonari Hirano and Hisato Kunitake

Plants 13 1864 2024.7

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

DOI： https://doi.org/10.3390/plants13131864

Effects of fertilization of male gametes with heavy-ion beam irradiation on embryo and endosperm development in Cyrtanthus mackenii Reviewed

Shii Makiko, Kajiya Yuki, Murata Muneaki, Abe Tomoko, Kunitake Hisato, Hirano Tomonari

CYTOLOGIA 89 ( 2 ) 133 - 139 2024.6

Language：English Publishing type：Research paper (scientific journal) Publisher：公益財団法人日本メンデル協会

Heavy-ion beams are widely used for mutagenesis. The type and size of the induced mutations vary depending on the ion species and velocity. DNA damage response of male gametes during pollen tube growth has been investigated using heavy-ion beam irradiation to the pollen of Cyrtanths mackerii (Amaryllidaceae), indicating that DNA damage induced by argon-ion beam is more difficult to repair than that by carbon-ion beam. In this study, we investigated the effects of DNA damage or mutations in the male gametes with argon-ion irradiation on double fertilization and subsequent embryo and endosperm development and compared the results with carbon-ion irradiation. In immature seeds after pollination with argon-ion-irradiated pollen grains at 10 Gy, there were two types of embryo sacs with embryo and endosperm and with egg cell/zygote and endosperm. The proportion of embryo sacs in the latter type was higher when using argon-ion-irradiated pollen than carbon-ion-irradiated pollen at 40 Gy, suggesting that qualitative differences between the two kinds of irradiation influenced embryo development. In many endosperms after pollination with the irradiated pollen, abnormal chromosome separation and enlarged endosperm nuclei were observed. This indicated that the chromosomal abnormalities in the irradiated male gametes were transmitted to the endosperm nuclei. The enlarged nucleus formation was higher in pollen irradiated with an argon-ion beam than in that with a carbon-ion beam. In conclusion, argon-ion beams, even at low doses, induce distinctive development of embryo and endosperm, making them valuable for investigating double fertilization.

DOI： 10.1508/cytologia.89.133

Changes in Carotenoids and Polyphenols during the Growth Stages of Orange-Fleshed Sweet Potato (Ipomoea batatas (L.) Lam.) Reviewed

Yuno Setoguchi 1, Yosuke Narasako, Tomonari Hirano, Motoyasu Otani and Hisato Kunitake 3

Horticulturae 10 629 2024.6

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

DOI： https://doi.org/10.3390/horticulturae10060629

Distinctive development of embryo and endosperm caused by male gametes irradiated with carbon-ion beam. Reviewed

Hirano T, Murata M, Watarikawa Y, Hoshino Y, Abe T, Kunitake H

Plant reproduction 2024.2

Authorship：Last author Language：English Publishing type：Research paper (scientific journal)

DOI： 10.1007/s00497-024-00496-9

Effect of Growth Stages on Anthocyanins and Polyphenols in the Root System of Sweet Potato. Reviewed

Setoguchi Y, Nakagawa S, Ohmura R, Toshima S, Park H, Narasako Y, Hirano T, Otani M, Kunitake H

Plants (Basel, Switzerland) 12 ( 9 ) 2023.5

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

DOI： 10.3390/plants12091907

Blueberry Stem Extract Prevents Lacrimal Hyposecretion in Non-obese Diabetic Mice via Activation of AMPK. Reviewed

Ogawa K, Ohno Y, Tagashira A, Urata K, Satoh K, Fujimoto N, Sonoda H, Ikeda M, Matsuzaki T, Nishiyama K, Kunitake H, Goto YO, Yamasaki M

In vivo (Athens, Greece) 37 ( 3 ) 1003 - 1015 2023.5

Language：English Publishing type：Research paper (scientific journal)

DOI： 10.21873/invivo.13174

Characterization of a novel mutant with inhibition of storage root formation in sweet potato Reviewed International coauthorship

Park Hyungjun, Abe Tomoko, Kunitake Hisato, Hirano Tomonari

Ｂｒｅｅｄｉｎｇ　Ｓｃｉｅｎｃｅ 22090 ( 0 ) 2023.4

Language：English Publishing type：Research paper (scientific journal) Publisher：日本育種学会

Sweet potato is a widely cultivated crop with storage roots. Although many studies have been conducted on the mechanism of its storage root formation, the details have not been fully elucidated. We screened mutant lines with inhibition of storage root formation to clarify parts of the mechanism. In this study, the process of storage root formation in one of the mutant lines, C20-8-1, was investigated. The inhibition of storage root formation was observed during the early stages of growth. The roots in C20-8-1 did not show histological differences compared to those in wild type. The transition from fibrous roots to pencil roots, which are the developmental stages prior to mature storage root formation, was delayed or inhibited in C20-8-1. The upregulation of starch biosynthesis-related genes and downregulation of lignin biosynthesis genes with storage root swelling were not confirmed in the root of C20-8-1 during the developmental transition stage, suggesting that most of the roots in C20-8-1 are in the pre-transition state toward the storage root swelling. C20-8-1 showed a mutant phenotype during the critical period of storage root swelling initiation, and further clarification of this mutation is expected to provide new insights into storage root formation.

DOI： 10.1270/jsbbs.22090

Amphidiploid production of hybrid between black raspberry and Rubus parvifolius L., a wild Asian species, using colchicine treatment Reviewed

Toshima S., Katsumi I., Kai A., Yahata M., Hirano T., Kunitake H.

Scientia Horticulturae 312 2023.3

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae

The fruit of the black raspberry (Rubus occidentalis L.) contains numerous bioactive components such as polyphenols, and the development of novel black raspberries with numerous functional components is eagerly desired. Rubus wild species in Asia, including Rubus parvifolius L., have been attracting a great deal of attention as sources of new traits in breeding raspberry and blackberry. In this study, we developed interspecific hybrids (BCP1 and BCP2) between black raspberry JP1 (Japanese accession 1) and the Asian Rubus parvifolius L. wild raspberry, and we confirmed interspecificity using simple-sequence repeat (SSR) markers. We then developed an amphidiploid using colchicine treatment and confirmed and evaluated it. Our results confirmed the interspecific hybridity of BCP1 and BCP2 using 4 SSR markers. In the colchicine treatment of BCP2, tetraploids were obtained after exposure to 0.1% colchicine for 2 days (7.7%) and 4 days (5.9%). The tetraploid of BCP2 showed vigorous growth, flowering, and fruiting in a warm region of Japan, and the flower diameters of BCP2 (4x) were significantly larger than those of BCP2 (2x). Also, the fruit color was confirmed to be black, the same as the fruit of JP1. We believe our research will support the development of black raspberry breeding and may contribute to ploidy breeding between black raspberries and wild Asian species as a new genetic resource for environmental adaptability.

DOI： 10.1016/j.scienta.2023.111863

Comparison of Proanthocyanidin Content in Rabbiteye Blueberry (Vaccinium virgatum Aiton) Leaves and the Promotion of Apoptosis against HL-60 Promyelocytic Leukemia Cells Using 'Kunisato 35 Gou' Leaf Extract. Invited Reviewed

Toyama Y, Fujita Y, Toshima S, Hirano T, Yamasaki M, Kunitake H

Plants (Basel, Switzerland) 12 4 2023.2

Authorship：Last author,　Corresponding author Language：English Publishing type：Research paper (scientific journal)

DOI： https://doi.org/10.3390/plants12040948

Blueberry stem extract and stem active components prevent blue light-emitting diode light-induced retinal photoreceptor cell damage in vitro. Reviewed

Ogawa K, Urata K, Suzuki Y, Sugamoto K, Goto Y, Nakayama T, Nishiyama K, Kunitake H, Yamasaki M

Bioscience, biotechnology, and biochemistry 87 ( 378 ) 388 2023.1

Language：English Publishing type：Research paper (scientific journal)

DOI： https://doi.org/10.1093/bbb/zbad001

Blueberry Leaf Extract Prevents Lacrimal Hyposecretion in Sjögren's Syndrome-like Model of Non-obese Diabetic Mice Reviewed

Ogawa K., Urata K., Maeda S., Ohno Y., Satoh K., Yamada Y., Suzuki Y., Koga Y., Sugamoto K., Kawaguchi M., Kunitake H., Nishiyama K., Goto Y., Nakayama T., Yamasaki M.

In vivo (Athens, Greece) 37 ( 1 ) 149 - 162 2023.1

Language：English Publishing type：Research paper (scientific journal) Publisher：In vivo (Athens, Greece)

BACKGROUND/AIM: This study evaluated the effect of blueberry leaf hot water extract (BLEx) on Sjögren's syndrome (SS)-like lacrimal hyposecretion in male non-obese diabetic (NOD) mice. MATERIALS AND METHODS: NOD or BALB/c mice were fed 1% BLEx or control (AIN-93G) for 2 weeks from the age of 4 to 6 weeks. Pilocarpine-induced tear volume was measured using a phenol red-impregnated thread. The lacrimal glands were evaluated histologically by H&E staining. The IL-1β and TNF-α levels in the lacrimal gland tissue were measured by ELISA. The mRNA expression levels of secretion-related proteins were measured by real-time PCR. LC3 I/II and arginase 1 expression levels were measured by western blot. RESULTS: After feeding with BLEx, pilocarpine-induced tear secretion in NOD mice was increased. In contrast, the mRNA expression levels of the cholinergic muscarinic M3 receptor, aquaporin 5, and ion channels related to lacrimal secretion were not changed by BLEx administration. In addition, the protein expression of arginase 1, which was recently reported to be involved in tear hyposecretion in NOD mice, was also not improved by BLEx administration. Although infiltration in the lacrimal gland of NOD mice was not decreased, the levels of TNF-α and the autophagy-related protein LC3 were significantly suppressed by BLEx treatment. CONCLUSION: BLEx treatment may ameliorate lacrimal hyposecretion in NOD mice by delaying the progression of autoimmune disease by suppressing autophagy in lacrimal glands.

DOI： 10.21873/invivo.13064

Blueberry Stem Extract Suppresses Blue Light-Emitting Diode Light-Induced Endoplasmic Reticulum Stress on Retinal Photoreceptor Cells

Ogawa Kenjirou, Karitani Takara, Otsu Wataru, Nishiyama Kazuo, Kunitake Hisato, Goto Yo, Nomiyama Shota, Hara Hideaki, Yamasaki Masao

BPB Reports 6 ( 3 ) 87 - 97 2023

Language：English Publishing type：Research paper (scientific journal) Publisher：公益社団法人日本薬学会

Background: Blue light causes retinal photoreceptor damage via oxidative and endoplasmic reticulum (ER) stress. A previous study showed that blueberry stem extract (BStEx) and its active components have cytoprotective effects against blue-light-induced photoreceptor cell damage by suppressing oxidative stress. This study demonstrated the inhibitory effect of BStEx against blue light-induced ER stress in photoreceptor cells. Methods: The photoreceptor cells treated with BStEx or the antioxidant N-Acetyl-L-cysteine (NAC) as a positive control were used and then exposed to blue light. The cytoprotective effects of BStEx and NAC were evaluated using CCK-8. The ER stress-related protein expression changes over time, and its levels were measured after each exposure time to blue light in photoreceptor cells treated with BStEx or NAC. Results: BStEx and NAC showed protective effects against blue-light-induced photoreceptor morphological abnormalities and cell damage. Although blue light triggered ER stress factors such as BiP, PERK, ATF6, eIF2α, ATF4, and CHOP, which in turn stimulated cell cycle arrest factors p53 and p21 and upregulation of apoptosis-inducing factors caspase-3. However, BStEx suppressed the increase in expression of BiP, ATF4, ATF6, CHOP, p53, p21, and caspase-3, but not mitochondrial apoptotic factors Bax and cytochrome c. Furthermore, the antioxidant NAC showed similar suppressive effects on BStEx. Conclusion: Our findings suggest that blue light-induced ER stress is primarily caused by oxidative stress, and BStEx might suppress ER stress via an antioxidant effect. The antioxidant NAC contributes to the cell proliferative capacity and suppression of apoptosis in photoreceptor cells.

DOI： 10.1248/bpbreports.6.3_87

Effects of In vitro Colchicine Treatment on Tetraploid Induction in Seeds of Polyembryonic Cultivars of the Genera Citrus, Fortunella, and Poncirus Invited Reviewed

YASUDA Kiichi, NUKAYA Tsunaki, SUDO Miki, YAHATA Masaki, TOMINAGA Akiyoshi, MUKAI Hiroo, KUNITAKE Hisato

Japanese journal of tropical agriculture 66 ( 2 ) 51 - 58 2022.9

Authorship：Last author Language：English Publishing type：Research paper (scientific journal) Publisher：日本熱帯農業学会

To produce tetraploid plants effectively, we examined the effects of colchicine treatment on tetraploid induction from seeds of five polyembryonic genotypes in Citrus, Fortunella, and Poncirus. In the ‘Valencia’ sweet orange (C. sinensis), the germination rate decreased with increasing colchicine-treatment concentration and/or duration. The Meiwa kumquat (F. classifolia) and Hongkong kumquat (F. hindsii) showed high germination rates (>90%) in all treatments for 24–72 h, regardless of the colchicine concentration, although their germination rate decreased with any 96 h colchicine treatment. There was no regularity in the germination rates of the trifoliate orange variation ‘Flying-dragon’ trifoliate orange (P. trifoliata) or the ‘Swingle’ citrumelo [grapefruit (C. paradis) x trifoliate orange]. The effects of the treatment concentration and duration on the germination rate and the number of total seedlings differed among genera. Although the seedlings obtained after colchicine-treatment showed various ploidy levels such as tetraploids, octaploids, and chimeras (2x + 4x, 4x + 8x), the tetraploid induction rate varied greatly with the plant materials. The tetraploid induction rate per seed in ‘Valencia’ was highest (53.3%) with the 24 h treatment with 0.1% colchicine. The tetraploid induction rates in the Meiwa (50.0%) and Hongkong (30.0%) kumquats were greatest with 72 h treatment with 0.2%. The ‘Flying-dragon’ trifoliate orange and ‘Swingle’ showed the highest tetraploid induction rates with treatment of 0.1% for 24 h (10.0%) and 0.2% for 24 h (7.3%), respectively. Compared to Citrus and Fortunella, the tetraploid induction rate in Poncirus and its hybrid was low, and no regularity was observed among all of the conditions.

DOI： 10.11248/jsta.66.51

Comprehensive effects of heavy-ion beam irradiation on sweet potato (Ipomoea batatas [L.] Lam.) Reviewed

Park Hyungjun, Narasako Yosuke, Abe Tomoko, Kunitake Hisato, Hirano Tomonari

Plant Biotechnology 39 ( 3 ) 311 - 316 2022.9

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Japanese Society for Plant Biotechnology

Sweet potato is a major root crop with nutritious tuberous roots. The mechanism of tuberous root development has not yet been adequately elucidated. Genetic resources are required to develop the molecular understanding of sweet potato. Heavy-ion beams were applied to hexaploid sweet potato for an increase in genetic variation, after which the comprehensive effects of heavy-ion beam irradiation were investigated. In vitro cultured shoots with an axillary bud of ‘Beniharuka’ were irradiated with Ar-ions at a dose of 1–5 Gy and C-ions at a dose of 5–20 Gy, and three irradiated lines were separated from each irradiated shoot. The shoot regeneration was inhibited at high doses of each ion irradiation. Ar-ion irradiation had an especially high biological effect on shoot regeneration. A total of 335 lines were obtained, consisting of 104 and 231 lines derived from Ar- and C-ion irradiation, respectively. The change in the DNA content of the lines was analyzed by flow cytometry to evaluate the irradiation-induced damage to the DNA. The two lines demonstrated significant differences in the DNA content and changes at the chromosome level. The screening for the morphological mutants was conducted in the field. Some irradiated lines showed inhibited or no tuberous root phenotype as mutant candidates. Additionally, the high-yield mutant candidates were dominated by Ar-ion irradiation. It was indicated that heavy-ion beam mutagenesis is effective in broadening the range of the phenotypes corresponding to tuberous root formation in hexaploid sweet potato.

DOI： 10.5511/plantbiotechnology.22.0725a

Highly polymerized proanthocyanidins (PAC) components from blueberry leaf and stem significantly inhibit SARS-CoV-2 infection via inhibition of ACE2 and viral 3CLpro enzymes Reviewed

Kazuhiro Sugamoto, Yuri L. Tanaka, Akatsuki Saito, Yoh Goto, Takayuki Nakayama, Tamaki Okabayashi, Hisato Kunitake, Kazuhiro Morishita

Biochemical and Biophysical Research Communications 615 56 - 62 2022.7

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier {BV}

DOI： 10.1016/j.bbrc.2022.04.072

Effects of cross combination on the total content and its composition of anthocyanins in sweetpotato (Ipomoea batatas L.) Reviewed

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Nakagawa S., Setoguchi Y., Ohmura R., Toshima S., Park H., Narasako Y., Hirano T., Otani M., Kunitake H.

Scientia Horticulturae 299 2022.6

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae

Anthocyanins in sweetpotato (Ipomoea batatas L.) have attracted much attention in light of their multiple health benefits and excellent properties as processed foods. Although crossbreeding is an effective way to breed new cultivars of sweetpotato, there are few reports on how anthocyanin content and composition are passed on to F1 hybrid generations by crossbreeding because of a complex genome and self-incompatibility reactions. In this study, three purple-fleshed sweetpotao cultivars, ‘Akemurasaki’, ‘Kyushu No.137’, ‘Purple Sweet Lord’, and one yellow-fleshed cultivars, ‘HE306’ (Silk Sweet®) selected for their anthocyanin contents and compositions were crossed in six combinations. Flowering was induced by using a sweetpotato grafting technique to obtain a total of 143 F1 hybrid lines. Anthocyanin content, the percentage of acylated anthocyanins, and the peonidin/cyanidin ratio in each of these F1 hybrid lines were investigated. In the F1 hybrid generations derived from these six cross combinations, many lines tended to have anthocyanin contents similar to those of the parent cultivars. In particular, among the 36 hybrid lines derived from crosses between ‘Akemurasaki’ and ‘HE306’ (Silk Sweet®), 13 lines with high anthocyanin contents (≧200 mg·100 g FW−1) and 20 lines without anthocyanins were identified. The F1 hybrid lines of this combination had anthocyanin contents similar to those of the parent cultivars, with anthocyanin-free lines appearing more frequently than anthocyanin-containing lines. Interestingly, we also identified a line with an extremely low percentage of acylated anthocyanins (49.7%) from the F1 hybrid lines derived from a cross between ‘Purple Sweet Lord’ (seed parent) and ‘Kyushu No. 137’ (pollen parent). Moreover, the F1 hybrid lines derived from crosses between ‘Purple Sweet Lord’ and ‘Kyushu No.137’ tended to have high peonidin/cyanidin ratios. This study provides an example of the inheritance of anthocyanin content and its composition in sweetpotato, which supplies valuable information for selection of parents to breed purple-fleshed cultivars.

DOI： 10.1016/j.scienta.2022.110999

Prediction of the Adult T-Cell Leukemia Inhibitory Activity of Blueberry Leaves/Stems Using Direct-Injection Electron Ionization-Mass Spectrometry Metabolomics. Invited Reviewed International journal

Hisahiro Kai, Yoshihito Okada, Yo Goto, Takayuki Nakayama, Kazuhiro Sugamoto, Kenjirou Ogawa, Masao Yamasaki, Kazuhiro Morishita, Koji Matsuno, Hisato Kunitake

Plants (Basel, Switzerland) 11 ( 10 ) 2022.5

Language：English Publishing type：Research paper (scientific journal)

Although Vaccinium virgatum Aiton leaves and stems inhibit adult T-cell leukemia (ATL) cells, leaves and stems can differ between individual plants and by time and location. In this study, leaf and stem components were profiled in the same individual plant using direct-injection electron ionization-mass spectrometry (DI-EI-MS) metabolomics, with the aims of analyzing the anti-ATL activity, and quantifying proanthocyanidins (PACs). Leaves, stems, and leaf/stem mixtures showed distinct and characteristic spectra. Anti-ATL activity was stronger in stems than leaves, and the PAC content was higher in stems than leaves. These data were subjected to bivariate analysis to identify the factor (m/z) responsible for the inhibitory effect of ATL based on the highest coefficient of determination (R2). The results of this DI-EI-MS metabolomics analysis suggest that among PACs contained in V. virgatum stems and leaves, the fragment ion at m/z 149 contributes significantly to anti-ATL activity.

DOI： 10.3390/plants11101343