1997 Fiscal Year Final Research Report Summary
Genetic transformation and various biotechnological treatments on hairy roots
| Project/Area Number |
08660003
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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 |
Breeding science
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| Research Institution | Chiba University |
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Principal Investigator |
MII Masahiro Faculty of Horticulture, Chiba University, Professor, 園芸部, 教授 (30093074)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Genetic engineering / Genetic transformation / Agrobacterium / Hairy roots / GUS gene / Mitotic synchronization / Polyploid production / Colchicine |
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| Research Abstract |
In this study, transformation of several horticultural plants with Agrobacterium rhizogenes was tried and the hairy roots thus obtained were used as the targets for the studies on the genetic transformation and various biotechnological studies. The results obtaiend are as follows :
1. Plant regeneration systems have been established from the hairy roots obtained by infection of leaf segments with wild-type strains of Agrobacterium rhizogenes in Nierembergia repens, N.acoparia, Ipomoea trichocarpa, snapdragon and grape vine.
2. Root tips of N.scoparia were infected with A.tumefaciens harboring GUS and NPT II genes. Hairy roots expressing GUS activity and resistance to kanamycin were selected and transgenic plants were regenerated from these hairy roots.
3. Root tip meristems were subjected with various treatments for mitotic synchronization and the highest mitotic index (30%) was obtained with the combination of 3 treatments including phosphate starvation, hydroxyurea and colchicine treatments.
4. Trials for isolating chromosomes from the synchronized root tip cells resulted in failure due to the break down of the chromosomos during the isolation process and difficulty in purification of the chromosomes.
5. Tetraploid plants were successfully regenerated from the hairy root cultures after the treatment with colchicine by adjusting the proper time and concentration.
6. These results indicate that root cultures including hairy roots are useful materials for genetic transformation and other biotechnological treatments.
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