Plant regeneration from petal protoplast culture ofPetunia hybrida |
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Authors: | Man-Ho Oh Sang-Gu Kim |
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Institution: | (1) Department of Biology, Seoul National University, 151-742 Seoul, Republic of Korea |
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Abstract: | Morphologically normal plants have been regenerated from petal protoplasts of petunia (Petunia hybrida) flower. Maximum protoplast yields from petal tissues were obtained within 2 days after anthesis. Protoplasts were cultured on modified Murashige and Skoog's medium in which NH4NO3 and Fe·EDTA concentrations were reduced to 1/3 (7mM) and 1/10 (10 M), respectively. After plating, protoplasts gradually reduced pigment density, and plastids developed near the nucleus. In premitotic petunia petal cells, the nucleus moved from the periphery to the central region of the cell. The first cell divisions were detected after 6–10 days of culture initiation, and the average division frequency was 15% in the best culture condition. The results indicated that the time of the first cell division and cell division frequency were closely related to flower age after anthesis. More than a hundred plants with morphologically normal shoots and roots have been obtained. Those plants grew vigorously in soil.Abbreviations BA
benzylaminopurine
- DAPI
4, 6-diamidino-2-phenylindole
- 2, 4-d
dichlorophenoxyacetic acid
- Fe·EDTA
Fe·ethylenediaminetetraacetate
- IAA
indole-3-acetic acid
- MtSB
microtubule stabilizing buffer
- NAA
-naphthaleneacetic acid
- PIPES
piperazine-N,N-bis(2-ethanesulfonic acid)
- EGTA
ethylene glycol-bis(-aminoethyl ether) N,N,N,N-tetraacetic acid |
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Keywords: | cell division DNA staining petal protoplasts Petunia hybrida regeneration |
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