| Project/Area Number |
61440010
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
園芸・造園学
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| Research Institution | Hokkaido University |
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Principal Investigator |
YAKUWA Toshiro Fac. of Agric., Hokkaido Univ., Professor, 農学部, 教授 (70001394)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Takashi Fac. of Agric., Hokkaido Univ. Assistant, 農学部, 助手 (30196836)
ARAKI Hajime Fac. of Agric., Hokkaido Univ. Assistant, 農学部, 助手 (30183148)
HARADA Takashi Fac. of Agric., Hokkaido Univ. Associate Professor, 農学部, 助教授 (30001457)
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| Project Period (FY) |
1986 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥28,000,000 (Direct Cost: ¥28,000,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1986: ¥20,000,000 (Direct Cost: ¥20,000,000)
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| Keywords | Protoplast culture / Somatic embryo formation / Embryo culture / Freeze preservation / Germplasm / Cell culture / Cell selection / Stem tip culture / 凍優保存 / 不定胚 / プロトプラスト / アスパラガス / ヤマノイモ属 / ヤマイモ属 / リンゴ / ブルーベリー |
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| Research Abstract |
(1)Culture of protoplasts : Plantlet regeneration from protoplasts derived from mesophyll cells of a true leaf of aseptically developed brussels sprout seedlings was successfully attained through in vitro culture system of colonization, callusing and somatic embryony.
(2)Somatic embryo formation Embryogenic calli, which were formed on the basal portion of the cultures (with elongated shoots) obtained through in vitro culture of shoot tips of an asparagus (Asparagus officinalis L.) spear, produced somatic embryos such as proembryos and globular embryos through shake culture; a few of the embryos developed into intact plants. Embryogenic caill were formed from a stem plate tissue(beneath apical meristem) and receptacle tissues of garlic, and from shoot tips of Yamatoimo(Dioscorea opposita T.) and immature embryos of Uchiwadokoro(Dioscorea nipponica M.).
(3)Production of hybrid plants by embryo culture : Dioscorea embryos (more than 0.6mm in diameter) cultured in vitro developed into hybrid
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plantlets. Even from immature embryos, borne by the hybridizations of D.opposita cv.Nagaimo x D.opposita cv.Nagaimo, D.japonica cv.jinenjo x D.opposita cv.Nagaimo, and D.opposita cv.Yamatolmo x D.opposita cv.Nagaimo, hybrid plants could be obtained through a method of culturing the embryos which were further matured by in vitro culture of seeds containing those immature embryos.
(4)Freeze-preservation of genetic resources or germplasms : In apples, storage of winter-excised dormant woods with a sufficient freezing resistance made it possible that plant materials (i.e., shoot tips) for freeze preservation could be obtained all the year round; thidiazuron, added to the media used for plantlet-regenerating culture after freezing and thawing enhanced shoot formation. Shoot tips of pears, grapes, cherries, edible lilies and Haskappu (Lonicera caerulea L. var.emphyllocalyx NAKAI) survived freezing in liquid nitrogen, and showed high survival rates.
(5)Establishment of mass propagation method : Shoot tip culture of Gyoja-nin-niku (Allium victorialis L. ssp.platyphyllum HULT), Haskappu (Lonicera caerulea L. var.emphyllocaryx NAKAI), blueberries, European pear induced shooting, followed by rooting, and regenerated intact plantlets.
(6)Cell culture : Whole plantlet regeneration of free cells isolated from asparagus (Asparagus officinalis L.)cladophylis was achieved by the cell-enclosing and colony-plating culture methods.
(7)Regeneration of disease-resistant plants by cell-selection method : calli resistant to asparagus stem blight were selected by the method that calli derived from asparagus (Asparagus officinalis L.) spear tissue were cultured using a medium containing phytotoxins related to the disease. The disease-resistant calli of three varieties have developed into plants. Less
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