| Project/Area Number |
05454042
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
作物学
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KIKUTA Yoshio Hokkaido Univ., Fac. of Agr., Pro., 農学部, 教授 (90001445)
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| Co-Investigator(Kenkyū-buntansha) |
FUJINO Kaien Hokkaido Univ., Fac. of Agr., Inst., 農学部, 助手 (80229020)
KODA Yasunori Hokkaido Univ., Fac. of Agr., A.Pro., 農学部, 助教授 (20002355)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1994: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1993: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | ABA-induced proteins / Embryogenesis / Gene expression / Gene-promotor / PKPI-gene / Potato tuberization / Rice long-term callus / Gene introduction / GUS活性 / Oryza sativa / Solanum tuberosum / Nicotiana tabacum / イネ種子カルス / 植物体再分化 / アブサイシン酸(ABA) / フォスフォリラーゼ / フォスフォグルコムターゼ / 硝酸還元酵素 / バレイショ / cDNAライブラリー / 貯蔵蛋白質 / プロテアーゼインヒビター / ジャスモン酸 / 日夜温較差 |
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| Research Abstract |
By using in vitro tuberization system of potato, the accumulation of potato kunitz type proteinase inhibitor (PKPI) mRNA was specifically detected in tubers during the early stage of development, and thus expected to be a molecular marker of potato tuberization. Furthermore, the accumulation of PKPI-mRNA was also found when tissues were treated with cool to warm atmosphere by ascending temperature, and by addition of jasmonic acid.
The novel proteins (14,18.5,24.5,45kDa) were induced by ABA in long-term cultured callus of rice and disappeared after plentlet regeneration, while proteins of the same molecular weight existed in intact mature seed embryos and disappeared when the seeds were germinating. Sustaining of embryogenic callus cultures could be some extent achieved by the addition of ABA in medium. This hormone could be responsible for regenerating somatic embryos and for normal embryogeny in rice plants.
A protocpl of the trangent gene expression in electroporated protoplasts has been developed for the introduction of foreign DNA into cells from cultured potato, tobacco, carrot, and rice. The method yields high amounts of the reporter enzyme B-glucuronidase (GUS) when cognate genes are driven by the promotors of NOS and 35S,and by upstream sequences of PKPI from potato tuber and Actin-1 from rice actin genes. Comparisons with expression in potato, tobacco, carrot, and rice cells indicate that trangent assays can be used to investigate promotor acivation and enhancer function.
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