2013 Fiscal Year Final Research Report
Development of genetic transformation of Japanese pear by inhibition of calcium ion influx
| Project/Area Number |
23658036
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Horticulture/Landscape architecture
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| Research Institution | National Agriculture and Food Research Organization |
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Principal Investigator |
NAKAJIMA Ikuko 独立行政法人農業・食品産業技術総合研究機構, 果樹研究所 栽培・流通利用研究領域, 主任研究員 (80355362)
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| Project Period (FY) |
2011 – 2012
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| Keywords | ニホンナシ / 子葉 / 再分化 / 形質転換 |
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| Research Abstract |
High rates of regeneration of adventitious shoots were obtained from the cotyledons of Japanese pears Imamuraaki and Agenosho Shinanashi from 16 Japanese pear cultivars, when the media contained 5 uM NAA combined with 10 or 25 uM BA were used. Cotyledons of Japanese pear cultivars were co-cultivated with Agrobacterium tumefaciens which contained a green fluorescent protein (GFP) gene. EGTA (a chelator of calcium) treatments and sonication were applied, which could prevent plant defense reaction and produce physical wounds across the tissue respectively. EGTA treatment did not show a positive effect on expression of GFP fluorescence whereas sonication significantly increased. One plant regenerated from Agenosho Shinanashi showed stable GFP fluorescence and confirmed as a transformant. Three other transformed regenerated shoots by myb gene showed red color, which were derived from Imamuraaki in another experiment. Transformation system in our study was shown to be reproducible.
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