| Project/Area Number |
60560037
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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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 | Saga University |
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Principal Investigator |
TASHIRO Yousuke Saga University, Faculty of Agriculture, Assistant Professor, 農学部, 助手 (20039340)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1986: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1985: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Protoplast / Cell fusion / 体細胞雑種 |
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| Research Abstract |
1. Protoplast isolation and culture
Different tissues from many vegetable crops were treated with a mixture of enzymes, including Cellulase Onozuka RS or R10 (1-1.5%) and Pectolyase Y-23 (0.1-0.2%) for 2.5 to 3 hours. This method generally resulted in high yields of protoplasts. Isolated protoplasts were cultured in various conditions. Cell divisions occurred in some protoplasts from most of the vegetable crops examined. Cell colonies were, however, formed only from the protoplasts from carrot and celery. Cell colonies from carrot protoplast were able to form many adventitious embryos after culturing in liquid media. These embryos formed many whole plants after culturing on agar solid media under light. condition.
2. Protoplast fusion and culture
Protoplasts from the vegetable crops belonging to Allium and Umbelliferae were successfully fused when they were treated with polyethylene glycol. The frequency of fusion was increased by enrichment of Ca and by increasing pH. Electro-fusion method were also effectual for the fusion of these protoplasts. Fusion products between carrot and celery protoplasts were cultured and formed vigorous cell colonies.
3. Identification and selection of somatic hybrids
By the karyotypic observations of cell colonies, adventitious embryos or plantlets from fusion products, heterokaryotic fusion products will be identified and selected.
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