Plant Cell Reports

, 24:572 | Cite as

Visual selection and maintenance of the cell lines with high plant regeneration ability and low ploidy level in Dianthus acicularis by monitoring with flow cytometry analysis

Cell Biology and Morphogenesis
First Online:
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Abstract

Efficient plant regeneration system from cell suspension cultures was established in D. acicularis (2n=90) by monitoring ploidy level and visual selection of the cultures. The ploidy level of the cell cultures closely related to the shoot regeneration ability. The cell lines comprising original ploidy levels (2C+4C cells corresponding to DNA contents of G1 and G2 cells of diploid plant, respectively) showed high regeneration ability, whereas those containing the cells with 8C or higher DNA C-values showed low or no regeneration ability. The highly regenerable cell lines thus selected consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.

Keywords

Plant regeneration Cell suspension culture Dianthus acicularis Burkholderia caryophylli Pseudomonas caryophylli Ploidy level Flow cytometry 

Abbreviations

Centrophenoxine

4-Chlorophenoxyacetic acid 2-dime-thylamino ethyl ester.

DAPI

4′, 6-Diamino-2-phenylindole dihydrochloride.

2,4-D

2,4-Dichlorophenoxyacetic acid.

FCM

Flow cytometry.

IAA

Indole-3-acetic acid.

NAA

α-Naphthaleneacetic acid.

PGR

Plant growth regulator.

Picloram

4-Amino-3,5,6-trichloropicolinic acid

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Laboratory of Plant Cell Technology, Faculty of HorticultureChiba UniversityMatsudo-CityJapan

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