Papers - KUNITAKE Hisato
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Abnormality of Gamete Formation in a Pummelo [Citrus maxima (Burm.) Merr.] Haploid
八幡 昌紀, 國武 久登, 安田 喜一, 平井 孝宜, 藪谷 勤, 山下 研介, 小松 春喜
Journal of the Japanese Society for Horticultural Science 80 ( 1 ) 14 - 18 2011
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：一般社団法人 園芸学会
半数体ブンタン［<i>C. maxima</i>（Burm.）Merr.］の雌性および雄性配偶子の形成過程を調査した．雌性配偶子形成では，胚のう母細胞が全く形成されないため，いずれの時期においても正常に発達した胚のうは全く観察されなかった．このことから，半数体ブンタンの雌性不稔は胚のう母細胞の無形成が原因であることが明らかとなった．一方，雄性配偶子形成では，半数体ブンタンは連続した 2 回の分裂を行っていたものの，ほとんどの分裂細胞で減数分裂の異常が認められた．一部の分裂細胞では，第一減数分裂後期においていくつかの一価染色体が両極に分配されず，赤道面付近に残るものや周りの細胞が第二減数分裂過程にもかかわらず，赤道面に 9 個の一価染色体が並列し，両極に分裂するものが観察された．さらに，花粉四分子期において，'晩白柚'では 99.3％が四分子であったのに対し，半数体ブンタンでは，一分子から六分子の小胞子型が観察され，二分子が 24.7％と，非常に高い頻度で出現していた．これらの結果は，半数体ブンタンの花粉が第一分裂復旧（FDR）のような異常によって稔性が回復していることを示唆した．<br>
DOI： 10.2503/jjshs1.80.14
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Nukaya Tsunaki, Ohta Tomohiro, Yasuda Kiichi, Yahata Masaki, Kunitake Hisato, Komatsu Haruki, Nii Naosuke, Mukai Hiroo, Harada Hisashi, Takagi Toshihiko
Hort. Res. (Japan) 10 ( 1 ) 1 - 8 2011
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE
Six lines of Meiwa kumquat (<i>Fortunella crassifolia</i> Swingle) showing tetraploidy (No.5, No.6, No.8, No.12, No.18 and No.19), which had been induced by treating the nucellar embryos with colchicines, were investigated for ploidy level, origin and horticultural characteristics. Furthermore, these tetraploid plants were crossed with diploid kumquats for triploid production. All tetraploid plants were confirmed to be derived from the nucellar embryo of Meiwa kumquat by investigation of chromosome composition with chromomycin A3 staining. Flow cytometric analysis and chromosome observation of young leaves demonstrated that No.8 was a ploidy chimera that was both diploid and tetraploid. Five lines showing tetraploidy had the typical morphological characteristics of tetraploid <i>Citrus</i> plants, such as large leaves, flowers and pollen grains with low pollen fertility. However, these tetraploid plants have desirable traits for kumquat such as thicker pericarp, high sugar content and smaller number of developed seeds. No.8 had horticultural characteristics that were similar to those of tetraploid plants excluding the dermal system. When their polyploids were crossed with some diploid cultivars, triploid progenies were obtained from almost all cross combinations.<br>
DOI： 10.2503/hrj.10.1
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Yahata, M., K. Yasuda, K. Nagasawa, S. Harusaki, H. Komatsu and H. Kunitake
Journal of the Japanese Society for Hortticultural Sciences 79 ( 3 ) 239 - 245 2010.9
Language：English Publishing type：Research paper (scientific journal)
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Investigation of sexual intergeneric haybrid Progenies between Citrus cultivar and Citropsis schweinfurthii
Yasuda, K., H. Kunitake, M. Yahata and R. Matsumoto
Proceedings of the International Society of Citriculture 1 121 - 125 2010.9
Language：English Publishing type：Research paper (international conference proceedings)
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Yahata M., Yasuda K., Nagasawa K., Harusaki S., Komatsu H., Kunitake H.
Journal of the Japanese Society for Horticultural Science 79 ( 3 ) 239 - 245 2010.8
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of the Japanese Society for Horticultural Science
To induce haploid plants in Citrus maxima (Burm.) Merr. 'Banpeiyu', we evaluated the effect of pollination with soft X-ray-irradiated pollen on fruit set and seed development, and carried out ovule culture. When 'Banpeiyu' pummelo pistils were pollinated with X-ray-irradiated pollen of 'Fukuhara' sweet orange [C. sinensis (L.) Osbeck], the exposure doses affected the fruit set. The number of seeds per fruit was also affected by the exposure dose, and tended to decrease as the dose increased; however, all developed seeds obtained from these crosses were diploid. In the ovule culture of 'Banpeiyu' pummelo, six embryoids shown haploidy were obtained in all treatments. One haploid plantlet with 9 chromosomes was regenerated from an embryoid in a culture of ovules established 40 days after pollination with 400 Gray (Gy)-irradiated pollen of 'Tosa-buntan' pummelo (C. maxima). This haploid was suggested to be derived from 'Banpeiyu' pummelo by random amplified polymorphic DNA (RAPD) and cleaved amplified polymorphic sequence (CAPS) analysis. JSHS © 2010.
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Yasuda K., Yahata M., Shigyo M., Matsumoto R., Yabuya T., Kunitake H.
Journal of the Japanese Society for Horticultural Science 79 ( 2 ) 129 - 134 2010.5
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of the Japanese Society for Horticultural Science
To obtain cytogenetic information on sexual intergeneric hybrid progenies between Citropsis schweinfurthii and 'Nanpu' tangor (Citrus spp.), we conducted Chromomycin A3 (CMA) chromosome banding and genomic in situ hybridization (GISH) analyses. The CMA karyotype composition of C. schweinfurthii was very simple (14D + 4E), while that of 'Nanpu' tangor showed more complicated chromosome types (1A + 4C + 5D + 8E). Two progenies with C. schweinfurthii as the maternal parent showed a karyotype of 1A + 1C + 11D + 5E, suggesting that each of the type A and C chromosomes was inherited from 'Nanpu' tangor. GISH-CMA analysis in one of these progenies showed a clear distinction between parental chromosomes, and revealed that the hybrid progeny arose due to fertilization between a female gamete with a CMA karyotype composition of 7D + 2E from C. schweinfurthii and a male 1A + 1C + 4D + 3E gamete from 'Nanpu' tangor. The result suggests that C. schweinfurthii had homomorphic homologous chromosomes as a wild species, while 'Nanpu' tangor had heteromorphic homologous chromosomes. © 2010 JSHS.
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Triploid and Aneuploid Hybrids from Diploid-Diploid Intergeneric Crosses between Citrus Cultivar ‘Kiyomi’ Tangor and Meiwa kumquat (Fortunella crassifolia Swingle) for Seedless Breeding of Kumquat.”jointly worked" Reviewed
「Yasuda K.」, 「M. Yahata」, 「H. Komatsu」,「Y. Kurogi」,「H. Kunitake」.
J Japan Soc Hort Sci 79 ( 2 ) 129 - 134 2010.4
Language：English Publishing type：Research paper (scientific journal)
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Yasuda K., Yahata M., Komatsu H., Kurogi Y., Kunitake H.
Journal of the Japanese Society for Horticultural Science 79 ( 1 ) 16 - 22 2010.2
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of the Japanese Society for Horticultural Science
In order to produce new seedless kumquat cultivars, we carried out an intergeneric cross between 'Kiyomi' tangor [Citrus unshiu Marcow. × C. sinensis (L.) Osbeck] and Meiwa kumquat (Fortunella crassifolia Swingle), obtaining 2 normal seeds and 7 undeveloped seeds. These seeds were cultivated on Murashige and Tucker medium, and the 2 normal seeds germinated and developed. The results of genome size analysis by flow cytometry revealed that both seedlings were triploids and that the difference in genome size corresponded to more than one chromosome in the 2 seedlings. Chromosome observation confirmed diploid (2n = 2x = 18) in both parents, aneuploid with 28 chromosomes (2n = 28) for one of the seedlings, and triploid (2n = 3x = 27) for the other seedling. Random amplified polymorphic DNA (RAPD) and cleaved amplified polymorphic sequence (CAPS) analyses proved that the seedlings were intergeneric hybrids between 'Kiyomi' tangor and Meiwa kumquat, with the maternal organelle genome. These hybrids have the potential to be released as a cultivar after further tree and fruit evaluations, and for use as cross-parents in seedless kumquat breeding. © 2010 JSHS.
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Phylogeny and classification of Fortunella (Aurantioideae) inferred from DNA polymorphism."jointly worked" Reviewed
「Yasuda K.」, 「M. Yahata」, 「H. Komatsu」, 「H. Kunitake」.
宮崎大学農学部研究報告 56 103 - 110 2010.2
Language：English Publishing type：Research paper (bulletin of university, research institution)
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Identification of Parental Chromosomes in Sexual Intergeneric Hybrid Progenies between Citrus Cultivar ‘Nanpu’ Tangor and Citropsis schweinfurthii in the Subfamily Aurantioideae. "jointly worked" Reviewed
「Yasuda, K」, 「M. Yahata」,「M. Shigyo」, 「R. Matsumoto」, 「T. Yabuya」, 「H. Kunitake」.
J Japan Soc Hort Sci 79 ( 1 ) 16 - 22 2010.1
Language：English Publishing type：Research paper (scientific journal)
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安田 喜一, 八幡 昌紀, 執行 正義, 松本 亮司, 藪谷 勤, 國武 久登
Journal of the Japanese Society for Horticultural Science 79 ( 2 ) 129 - 134 2010
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：一般社団法人 園芸学会
本研究では，カンキツ栽培品種'南風'タンゴールと <i>Citropsis schweinfurthii</i> 間の有性属間雑種における細胞遺伝学的知見を得るために，Chromomycin A3（CMA）核型分析と genomic <i>in situ</i> hybridization（GISH）を行った．その結果，<i>C. schweinfurthii</i> の CMA 核型構成が 14D + 4E と単純であったのに対し，'南風'タンゴールの CMA 核型構成は 1A + 4C + 5D + 8Eと複雑であった．また，<i>C. schweinfurthii</i> を種子親とする雑種 2 個体の CMA 核型構成はともに 1A + 1C + 11D + 5E であり，これらの A 型と C 型染色体は'南風'タンゴール由来のものと考えられた．さらに，その 1 個体について GISH-CMA 分析を行った結果，両親の染色体を明白に識別することができ，<i>C. schweinfurthii</i> 由来の雌性配偶子 7D + 2E と'南風'タンゴール由来の雄性配偶子 1A + 1C + 4D + 3Eとの受精によることが明らかになった．この結果から，野生種である <i>C. schweinfurthii</i> は同型の CMA 核型が相同染色体として対合するのに対し，'南風'タンゴールは，異型の CMA 核型染色体が対合することが示された．<br>
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安田 喜一, 八幡 昌紀, 小松 春喜, 黒木 義一, 國武 久登
Journal of the Japanese Society for Horticultural Science 79 ( 1 ) 16 - 22 2010
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：一般社団法人 園芸学会
キンカンの新しい無核性品種育成のために，'清見'タンゴール［<i>Citrus unshiu</i> Marcow. × <i>C. sinensis</i> (L.) Osbeck］とニンポウキンカン（<i>Fortunella crassifolia</i> Swingle）間の属間交雑を行った結果，2 個の完全種子と 7 個の不完全種子が得られた．交雑から得られた種子を MT 培地上で培養したところ，2 個の完全種子のみが発芽し，植物体へと成長した．まず，これらの実生の倍数性を確認するために，フローサイトメーターを用いたゲノムサイズ分析を行った．両実生は，ともに三倍性のピークを示したものの，それらの間には染色体 1 本以上に相当するゲノムサイズの差がみられた．このため，染色体の観察を行ったところ，一方の実生が 27 本の染色体を有する三倍体であったのに対し，他方は 28 本の染色体を有する異数体であることが明らかになった．次に，RAPD 分析を行った結果，両実生は両親に特異的なバンドを有しており，雑種であることが確認された．また，オルガネラ DNA 領域の CAPS 分析によって，両雑種が'清見'タンゴールのオルガネラゲノムを有することを確認した．これらの雑種は，すでに着花・結実に至っており，今後キンカンの新品種もしくは無核性品種育成のための中間母本となることが期待される．<br>
DOI： 10.2503/jjshs1.79.16
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八幡 昌紀, 安田 喜一, 長澤 幸治, 春崎 聖一, 小松 春喜, 國武 久登
Journal of the Japanese Society for Horticultural Science 79 ( 3 ) 239 - 245 2010
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：一般社団法人 園芸学会
ブンタン［<i>Citrus maxima</i> （Burm.）Merr.］'晩白柚'の半数体を誘導するために，軟 X 線照射花粉を受粉して着果と種子発育に及ぼす影響を調査するとともに，胚珠培養を行った．軟 X 線を照射した'福原'スイートオレンジ［<i>C. sinensis</i> （L.）Osbeck］の花粉を'晩白柚'に受粉すると，着果率はやや低下した．また，1 果実当たりの種子数は照射量の増加に伴い減少した．しかしながら，これらの交雑から得られた実生はすべて二倍体であった．一方，軟 X 線を照射した'福原'スイートオレンジと'土佐文旦'（<i>C. maxima</i>）花粉を受粉して胚珠培養を行うと，6 系統の半数性を示す胚様体が誘導された．その内，400 Gray 照射した'土佐文旦'花粉を受粉後 40 日に胚珠培養した胚様体から染色体数 9 本を有する半数体植物が 1 個体得られ，RAPD および CAPS 解析によりこの半数体は'晩白柚'由来であることが示された．<br>
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Investigation of sexual intergeneric hybrid progenies between Citrus cultivar and Citropsis schweinfurthii. Reviewed
Yasuda K., Kunitake, H., Yahata, M., Matsumoto, R.
Proceedings of the international society of citriculture 1 121 - 125 2009.9
Language：English Publishing type：Research paper (international conference proceedings)
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Fruit characteristics in autotetraploid meiwa kumquat induced by colchicine treatment to nucellar embryos. Reviewed
T. Nukaya, M. Yahata, K. Suzuki, K. Yasuda, H. Kunitake, H. Komatsu, H. Mukai, H. Harada, T. Takagi
Korean society for horticultural science 50 ( 3 ) 188 - 190 2009.9
Language：English Publishing type：Research paper (scientific journal)
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Evaluation of basal media for micropropagation of four highbush blueberry cultivars. Reviewed
Tetsumura, T., Y. Matsumoto, M. Sato, C. Honsho, K. Yamashita, H. Komatsu, Y. Sugimoto, H. Kunitake
Scientia Horticulturae 119 ( 1 ) 72 - 74 2008.12
Language：English Publishing type：Research paper (scientific journal)
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Evaluation of basal media for micropropagation of four highbush blueberry cultivars
Tetsumura T., Matsumoto Y., Sato M., Honsho C., Yamashita K., Komatsu H., Sugimoto Y., Kunitake H.
Scientia Horticulturae 119 ( 1 ) 72 - 74 2008.12
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Scientia Horticulturae
Murashige and Skoog medium (MS), woody plant medium (WPM), and a mixture of equal parts of MS and WPM (MW) were compared for in vitro shoot proliferation and rooting of four highbush blueberry (Vaccinium spp.) cultivars. During the multiplication stage, the shoots on WPM showed worse growth than the other shoots. The MW produced the best shoot growth. The shoots on MS grew well but tended toward hyperhydricity in the multiplication stage, especially in the case of 'Bluecrop' shoots. Rooting was worst in the shoots multiplied on WPM, while the best rooting percentage of 'Bluecrop' shoots was obtained on MS, and that of 'O'Neal' on MW. The rooted shoots developed a good root system and were easily acclimatized after potting. © 2008 Elsevier B.V. All rights reserved.
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The confirmation of ploidy periclinal chimera and its morphological characteristics in Meiwa kumquat `Yubeni.' Reviewed
Yasada K., H. Kunitake, S. Nakagawa, H. Kuroki, M. Yahata, R. Hirata, Y. Yoshikura, I. Kawakami, Y. Sugimoto
Horticultural Research 7 ( 2 ) 165 - 171 2008.4
Language：Japanese Publishing type：Research paper (scientific journal)
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Yasuda Kiichi, Kunitake Hisato, Nakagawa Syoko, Kurogi Hironori, Yahata Masaki, Hirata Rikiya, Yoshikura Yukihiro, Kawakami Ikuo, Sugimoto Yasuhiro
Hort. Res. (Japan) 7 ( 2 ) 165 - 171 2008
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE
The cultivar 'Yubeni' was discovered as a mutant of Meiwa kumquat (<i>Fortunella crassifolia</i> Swingle) in Kushima city, Miyazaki. According to the results of flow cytometry analysis and morphological investigation of several tissues and organs, 'Yubeni' was supposed to be a ploidy periclinal chimera with diploid (2n = 2X = 18) in the outermost layer (histogenic layer L-I) and tetraploid (2n = 4X = 36) inner layer (histogenic layer L-II and L-III) in the shoot apical meristem. Compared with the diploid line, it showed similar morphology to the tetraploid line such as larger leaves, flowers and pollen grains. The weight and size of the fruit were both greater than those of the diploid line. Furthermore, this cultivar has desirable traits for kumquat such as smaller number of developed seed and thicker pericarp.<br>
DOI： 10.2503/hrj.7.165
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Yasuda Kiichi, Kunitake Hisato, Nakagawa Syoko, Kurogi Hironori, Yahata Masaki, Hirata Rikiya, Yoshikura Yukihiro, Kawakami Ikuo, Sugimoto Yasuhiro
Horticultural research (Japan) 7 ( 2 ) 165 - 171 2008
Language：Japanese Publishing type：Research paper (scientific journal)
The cultivar 'Yubeni' was discovered as a mutant of Meiwa kumquat (Fortunella crassifolia Swingle) in Kushima city, Miyazaki. According to the results of flow cytometry analysis and morphological investigation of several tissues and organs, 'Yubeni' was supposed to be a ploidy periclinal chimera with diploid (2n = 2X = 18) in the outermost layer (histogenic layer L-I) and tetraploid (2n = 4X = 36) inner layer (histogenic layer L-II and L-III) in the shoot apical meristem. Compared with the diploid line, it showed similar morphology to the tetraploid line such as larger leaves, flowers and pollen grains. The weight and size of the fruit were both greater than those of the diploid line. Furthermore, this cultivar has desirable traits for kumquat such as smaller number of developed seed and thicker pericarp.