2002 Fiscal Year Final Research Report Summary
Shoot differentiation and gene-transfer in Periploca sepium Bunge
| Project/Area Number |
13660148
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
林学
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| Research Institution | Kyushu University |
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Principal Investigator |
GYOKUSEN Koichiro Kyushu University, Faculty of Agriculture, Associate Prof., 農学研究院, 助教授 (80205062)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Periploca / Tree / Shoot differentiation / Polyisoplene / Gene-transfer / Agrobacterium method / GFP / Ferritin |
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| Research Abstract |
Cotyledon and hypocotyl of Periploca sepium Bunge, which is a native liana tree species and contents rubber element in china, were used for shoot differentiation. MS-medium was used and both of BAP and NAA are used as hormones for shoot differentiation. Shoot differentiation was recognized in both cotyledon and hypocotyls, but differentiation rate was higher in hypocotyl. Root differentiation was recognized by using 1/2MS with 6μ mol/l IAA.
Stem and leaf of Periploca were used for shoot differentiation. MS-medium was used and both of BAP and NAA were added as hormones for shoot differentiation. Differentiation rate in stem was higher in the combination of BAP 1μM〜3μM and NAA 0.1μM and the combination of BAP 3μM〜10μM and NAA 0.1〜1μM in leaf. High differentiation rate was recognized and that was in the range from 90% to 100%.
Agrobacterium-mediated gene-transfer method was introduced for Periploca. GFP gene was used as a model-gene and stem from micropropagated shoot was used for explant. A plantlet that had green fluorescent under microscope was regenerated from a stem. PCR analysis and Western Analysis were used to investigate the DNA and GFP protein. From the result of both analysis, gene transfer of GPF gene had succeeded in Periploca.
Agrobacterium-mediated gene -transfer method was introduced to Ferritin gene. Two plantlets had been regenerated from shoot stem, and DNA segment was detected in these two plantlets by PCR analysis.
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