Fine mapping and cloning of apomixis genes in guineagrass using HEGS-DD methods

• Project/Area Number: 17380005
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Breeding science
• Research Institution: Minami Kyusyu University (2007-2008); University of Miyazaki (2005-2006)
• Principal Investigator: CHEN Lanzhuang Minami Kyusyu University, 園芸学部, 教授 (40284822)
• Co-Investigator(Kenkyū-buntansha): KAWASAKI Shinji 独立行政法人農業生物資源研究所, 生理機能研究グループ, 上席研究員 (70399426) ; TERAO Hiroyuki 宮崎大学, 農学部, 准教授 (60117174)
• Co-Investigator(Renkei-kenkyūsha): MATSUOKA Hidemichi 独立行政法人九州・沖縄農業研究センター, 畜産飼料作物研究部, 上席研究官 ; SUGITA Toru 宮崎県農業総合試験場, 生物工学部, 主任技師
• Research Collaborator: GUAN Liming 宮崎大学, 非常勤講師
• Project Period (FY): 2005 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥17,560,000 (Direct Cost: ¥15,700,000、Indirect Cost: ¥1,860,000); Fiscal Year 2008: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2007: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2005: ¥6,000,000 (Direct Cost: ¥6,000,000)
• Keywords: アポミクシス / ギニアグラス / HEGS-AFLP / HEGS-DD / アポミクシス遺伝子マッピング / アポミクシス遺伝子クローニング / 交配実験 / RNA・cDNA / 遺伝子マッピング / 遺伝子クローニング / 組換えイネ / 雄性不稔性 / RNA抽出

Research Abstract

本研究は、イネ科牧草ギニアグラスを実験材料とし、HEGS-AFLPとHEGS-DD法を用いてアポミクシス遺伝子のクローニングとそのマッピングを行ったものである。その成果、(1)アポミクシス遺伝子を4つ捕捉でき、異なる塩基配列長をもつ同じASG-1遺伝子ファミリーであることが明らかになった。(2)ASG-1を用いてイネ遺伝子組換え植物を効率的に作出でき、結実しないイネを作出した。(3)有性生殖とアポミクシス系統との交配で作ったF1集団を利用してアポミクシス遺伝子の精密マップを作製した。

Research Products

• [Journal Article] Ultrastructural studies of embryo abortion in buckwheat (Fagopyrum esculentum) as a heat-stress. (2008)
  • Author(s): Guan, L.M., L.Z. Chen, T. Adachi.
  • Journal Title: Cytologia 73 Pages: 371-379
  • NAID: 10024883555
  • Peer Reviewed
• [Journal Article] Clonal propagation and quick detection of virus-free plants of sweet potato, Ipomoea batatas. (2008)
  • Author(s): Chen, L.Z., Z. S. Du, T. Hamaguchi, T. Sugita, R. Nagata, H. Terao and T. E. Tsuzuki
  • Journal Title: Bull. Minamikyushu Univ. 38 Pages: 1-5
  • NAID: 40016068149
  • Peer Reviewed
• [Journal Article] Ultrastructural studies of Gametophytic Apomicts in Guinea grass (Panicum maximum) II. Characteristics of aposporous initial cell-derived embryo sac. (2008)
  • Author(s): Guan, L.M., L.Z. Chen, H. Terao
  • Journal Title: Cytologia 72
  • NAID: 10019016091
  • Peer Reviewed
• [Journal Article] Ultrastmctural studies of embryo abortion in buckwheat (Fagopyrum esculentum)as a heat-stress. (2008)
  • Author(s): Guan, LM, Chen, LZ, Adachi, T.
  • Journal Title: Cytologia 73(4) Pages: 371-379
  • Peer Reviewed
• [Journal Article] Clonal propagation and quick detection of virus-free plants of sweet potopo, Ipomoea batatas. (2008)
  • Author(s): Chen, LZ, Du, ZS, Hamaguchi, T, et al.
  • Journal Title: Bulletion of Minami Kyushu University 38A Pages: 1-5
  • Peer Reviewed
• [Journal Article] Clonal propagation and quick detection of virus-free plants of sweet potato,Ipomoea batatas (2008)
  • Author(s): Chen LZ, Z Du, T.Hamaguchi, T Sugita, R Nagata, H Terao, E Tsuzuki
  • Journal Title: Bulletin of Minami Kyushu University 38A Pages: 1-5
  • NAID: 40016068149
  • Peer Reviewed
• [Journal Article] Ultrastructural studies of gametophytic apomicts in guinea grass (Panicum maximum). I. Differentiation of aposporous initial cell. (2007)
  • Author(s): Guan, L.M., L. Z. Chen and H. Terao
  • Journal Title: Cytologia 71 Pages: 379-389
  • NAID: 10018287501
  • Peer Reviewed
• [Journal Article] Ultrastructural studies of gametophytic apomicts in guinea grass(Panicum maximum)II,Characteristics of aposporous initialcell-derived embryo sac (2007)
  • Author(s): Guan LM and LZ Chen
  • Journal Title: Cytologia 72 Pages: 145-153
  • Peer Reviewed
• [Journal Article] Preliminary isolation of viable embryo sacs and protoplasts from viable egg cells in facultative apomictic guineagrass (Panicum maximum). (2006)
  • Author(s): Chen, L.Z., S. Amano, L.M. Guan and T. Adachi
  • Journal Title: Cytologia 70 Pages: 171-179
  • Peer Reviewed
• [Journal Article] Ultrastructural studies of Gametophytic Apomicts in Guinea grass (Fanicum maximum) I. Differentiation of aposporous initial cell (2006)
  • Author(s): Liming GUAN, Lanzhuan CHEN, Hiroyuki TERAO
  • Journal Title: Cytologia 71 Pages: 379-389
• [Journal Article] Rapid construction of a sweet pepper genetic linkage map using high-efficiency genome scanning/AFLP and RAPD, from an intraspecific, doubled-haploid sweet pepper population. (2005)
  • Author(s): Sugita, T., T. Kinoshita, T. Kawano, K. Yuji, K. Yamagichi, R. Nagata, A. Shimizu, L.Z. Chen, S. Kawasaki, A. Todoroki
  • Journal Title: Breeding Science 55 Pages: 287-296
  • Peer Reviewed
• [Journal Article] Identification of a novel thyroidhormone-sulfating cytosolic sulfotransferase, SULT1 ST5, from zebrafish :Molecular cloning, expression, characterization and ontogenic study. (2005)
  • Author(s): Yasuda, S. AP. Kumar, M-Y. Liu, Y. Sakakibara, M. Suiko, L.Z. Chen, M-C. Liu
  • Journal Title: FEBS Journal(Formerly European Journal of Biochemistry) 272 Pages: 3828-3837
  • Peer Reviewed
• [Journal Article] Developmental expression of ASG-1 during gametogenesis in apomictic guinea grass (Panicum maximum). (2005)
  • Author(s): Chen, L.Z., L.M. Guan, M.K. Seo, F. Hoffmann and T. Adachi
  • Journal Title: Journal of Plant Physiology 162 Pages: 1141-1148
  • Peer Reviewed
• [Journal Article] Identification of a novel estrogen-sulfating cytosolic SUIT from zebrafish: Molecular cloning, expression, characterization, and ontogeny study. (2005)
  • Author(s): Yasuda, S., C-C. Liu, S. Takahashi, M. Suiko, L.Z. Chen, R. Snow and M-C. Liu
  • Journal Title: Biochemical and Biophysical Research Communications 330 Pages: 219-225
  • Peer Reviewed
• [Journal Article] Preliminary isolation of viable embryo sacs and protoplasts from viable egg cells in facultative apomictic guineagrass (Panicum maximum). (2005)
  • Author(s): Chen, L.Z., S.Amano, L.M.Guan, T.Adachi
  • Journal Title: Cytologia, 70 Pages: 171-179
• [Journal Article] Developmental expression of ASG-1 during gametogenesis in apomictic guinea grass (Panicum maximum). (2005)
  • Author(s): Chen, L.Z., L.M.Guan, M.K.Seo, F.Hoffmann, T.Adachi
  • Journal Title: Journal of Plant Physiology, 162 Pages: 1141-1148
• [Journal Article] Identification of a novel estrogen-sulfating cytosolic SUIT from zebrafish : Molecular cloning, expression, characterization, and ontogeny study. (2005)
  • Author(s): Yasuda, S., C-C.Liu, S.Takahashi, M.Suiko, L.Z.Chen, R.Snow, M-C.Liu
  • Journal Title: Biochemical and Biophysical Research Communications, 330 Pages: 219-225
• [Journal Article] Rapid construction of a sweet pepper genetic linkage map using high-efficiency genome scanning/AFLP and RAPD, from an intraspecific, doubled-haploid sweet pepper population. (2005)
  • Author(s): Sugita, T., T.Kinoshita, T.Kawano, K.Yuji, K.Yamagichi, R.Nagata, A.Shimizu, L.Z.Chen, S.Kawasaki, A.Todoroki
  • Journal Title: Breeding Science, 55 Pages: 287-296
• [Journal Article] Identification of a novel thyroidhormone-sulfating cytosolic sulfotransferase, SULT1 ST5, from zebrafish : Molecular cloning, expression, characterization and ontogenic study. (2005)
  • Author(s): Yasuda, S., AP.Kumar, M-Y.Liu, Y.Sakakibara, M.Suiko, L.Z.Chen, M-C.Liu
  • Journal Title: FEBS Journal (Formerly European Journal of Biochemistry) 272 Pages: 3828-3837
• [Journal Article] Efficient Agrobacterium -mediated transformation of ASG-1 gene from guinea grass (Panicum maximum) to rice. (2004)
  • Author(s): Seo, M.K., L.Z. Chen, H.Y. Lee and T. Adachi
  • Journal Title: Applied Biological Science 8 Pages: 34-46
  • Peer Reviewed
• [Journal Article] Isolation and characterization of apomixis- specific gene family with different length of sequences in facultative apomictic guinea grass (Panicum maximum): Is it the evidence supporting hypothesis of 1 dominant gene of apomixis?
  • Author(s): Chen, L.Z., Guan, L.M., Miyazaki, C., Kojima, A., Saito, A.
  • Journal Title: Journal of Plant Physiology (in press)
  • Peer Reviewed
• [Journal Article] Ultrastructural studies of Gametophytic Apomicts in Guinea grass (Panicum maximum) II. Characteristics of aposporous initial cell-derived embryo sac
  • Author(s): Liming GUAN, Lanzhuan CHEN, Hiroyuki TERAO
  • Journal Title: Cytologia 72(in press)
  • NAID: 10019016091
• [Presentation] ギニアグラスにおけるアポスポリー性胚嚢始原細胞出現の微細解析 (2009)
  • Author(s): 陳 蘭荘・関 黎明
  • Organizer: 日本育種学会 第115回講演会 (育種学研究 第11巻別冊1号 p222)
• [Presentation] ギニアグラスにおけるアポスポリー性胚嚢始原細胞出現の微細解析 (2009)
  • Author(s): 陳蘭荘, 関黎明
  • Organizer: 日本育種学会台115回講演会
  • Place of Presentation: つくば市
• [Presentation] Apomixis specific gene-1 (ASG-1) isolated from Panicum maximum expressing pollen sterile in transgenic rice (2008)
  • Author(s): Chen, L.Z., L.M. Guan, T. Sugita, H. Ichikawa
  • Organizer: Abstracts of The International Symposium of Rice Functional Genomics. Pp298,
  • Place of Presentation: Jeju Korea
• [Presentation] イネ科アポミクシス性ギニアグラスの自家不和合性のメカニズム (2008)
  • Author(s): 陳蘭荘・関黎明・杉田亘・市川裕章・浜口卓郎
  • Organizer: 日本育種学会 第113回講演会 (育種学研究 第10巻別冊1号 p279)
• [Presentation] Apomixis specific gene-1 isolated from Panicum maximum expressing pollen sterile in transgenic rice. (2008)
  • Author(s): Chen, LZ, Guan, LM, Sugita, T., et al.
  • Organizer: The 6th International symposium of rice functional genomics
  • Place of Presentation: JEJU, KOREA
• [Presentation] イネ科アポミクシス性ギニアグラスの自家不和合性のメカニズム。(第10巻別冊1号p279) (2008)
  • Author(s): 陳 蘭荘・関 黎明・杉田 亘・市川 裕章・濱口 卓郎
  • Organizer: 日本育種学会 第113回講演会 育種学研究
  • Place of Presentation: 明治大学
• [Presentation] ギニアグラスにおけるアポスポリー性アポミクシス特異的遺伝子群の単離とその機能解析:アポミクシス1優性遺伝子支配説を支持する証拠? (2007)
  • Author(s): 陳蘭荘・関黎明・宮崎力・小島昭夫・斉藤彰
  • Organizer: 日本育種学会 第112回講演会 (育種学研究 第9巻別冊2号 p304)
• [Presentation] アポミクシス関連遺伝子ASG-1が組換えイネで発現する (2007)
  • Author(s): 陳蘭荘・関黎明・浜口卓郎・杉田亘・市川裕章
  • Organizer: 日本育種学会 第111回講演会 (育種学研究 第9巻別冊1号 p254)
• [Presentation] ギニアグラスにおけるアポスポリー性アポミクシス特異的遺伝子群の単離とその機能解析:アポミクシス1優性遺伝子支配説を支持する証拠(第9巻別冊2号p304) (2007)
  • Author(s): 陳 蘭荘・関 黎明・宮崎 力・小島 昭夫・斉藤 彰
  • Organizer: 日本育種学会 第112回講演会 育種学研究
  • Place of Presentation: 山形大学
• [Presentation] Aposporous apomixis may be controlled by single gene with different length of sequence in Panicum maximum. (2006)
  • Author(s): Chen, L.Z., L.M. Guan
  • Organizer: Abstracts of 8^<th> International Congress of Plant Molecular Biology
  • Place of Presentation: Adelaide, Australia
• [Presentation] Plant regeneration and functional analysis of ASG-1 transgenic guineagrass and rice plants mediated with Agrobacterium (2006)
  • Author(s): Chen, L.Z., L.M. Guan
  • Organizer: Abstracts of Tropical Crop Biotechnology Conference 2006
  • Place of Presentation: Far North Queensland city of Cairns, Australia,
• [Presentation] HEGS法を用いたアポミクシス遺伝子の精密マッピング:胚嚢分析と系統樹について (2006)
  • Author(s): 陳蘭荘・井上公一・森山聡・杉田亘・川崎信二・寺尾寛之
  • Organizer: 日本育種学会 第110回講演会 (育種学研究 第8巻別冊2号 p324)
• [Book] Chapter 7 Cell differentiation research trade: The mechanism of sexual cell differentiation in cultures of protoplasts, interspecific hybrid embryos and asexual tissues. In [Cell Differentiation Research Developments] Laura B. Ivanova (Ed) (2008)
  • Author(s): LZ Chen, and LM Guan.
  • Publisher: NOVA Science Publishers USA
• [Book] Chapter 7-Cell Differentiation Research Trends: The mechanism of asexual cell differentiation in cultures of protoplasts, interspecific hybrid embryos and asexual tissues, pp171-182. In [Cell Differentiation Research Developments] Laura B. Ivanova (Editor) (2007)
  • Author(s): L.Z. Chen, L.M. Guan
  • Publisher: Nova Science Publishers,
• [Book] Somatic embryogenesis, plant regeneration and clonal propagation in different tissue sources of different flowering plants - in monocots and dicots. Pp299-304. In [Floriculture, ornamental and plant biotechnology: Advances and topical issues] (1st Edition) Jaime A Teixeira da Silva (Ed) (2006)
  • Author(s): L.Z. Chen, L.M. Guan and Jaime A Teixeira da Silva
  • Publisher: Global Science Books,
• [Book] Advances in cloning and expression of apomixis-specific genes in flowering plants. Pp572-577. In Floriculture, ornamental and plant biotechnology: Advances and topical issues] (1st Edition) Jaime A Teixeira da Silva(Ed) (2006)
  • Author(s): L.Z. Chen and L.M. Guan
  • Publisher: Global Science Books,
• [Book] Floriculture, Ornamental and Plant Biotechnology Advances and Topical Issues I (ed. Jaime A. Teixeira da Silva) Advances in Cloning and Expression of Apomixis-Specific Genes in Flowering Plants (2006)
  • Author(s): LZ Chen, LM Guan
  • Publisher: Global Science Books, Ltd. UK
• [Book] Floriculture, Ornamental and Plant Biotechnology Advances and Topical Issues II (ed. Jaime A. Teixeira da Silva) Somatic embryogenesis, Plant Regeneration and Clonal Propagation in Different Tissue Sources of Different Flowering Plants : Monocots and Dicots (2006)
  • Author(s): LZ Chen, LM Guan, Jaime A, Teixeira da Silva
  • Publisher: Global Science Books, Ltd. UK
• [Remarks] 平成18年度 宮崎県文化賞(学術部門) 受賞内容「アポミクシス研究を通じて作物の品種改良に寄与するとともに、宮崎県の学術レベルの向上に貢献した」