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Increasing ploidy level in cell suspension cultures of Doritaenopsis by exogenous application of 2,4‐dichlorophenoxyacetic acid
Author(s) -
Mishiba Keiichiro,
Okamoto Tomonori,
Mii Masahiro
Publication year - 2001
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1034/j.1399-3054.2001.1120119.x
Subject(s) - biology , flow cytometry , cell cycle , ploidy , mitosis , cell culture , endoreduplication , cell , tissue culture , 2,4 dichlorophenoxyacetic acid , cell growth , population , cell division , subculture (biology) , picloram , plant cell , botany , microbiology and biotechnology , biochemistry , in vitro , genetics , demography , sociology , gene
To clarify the causal factors for ploidy variation in plant cell culture, we attempted to alter ploidy distribution in cell cultures of a tetraploid cultivar of Doritaenopsis by changing the plant growth regulators (PGRs) in the culture medium. The original suspension cultured cells, which had been maintained in medium containing 0.1 mg l −1 1‐naphthaleneacetic acid and 1 mg l −1 benzyladenine, were transferred onto various gellan gum solidified media with a single application of PGRs, and the ploidy distributions of the cells were examined using flow cytometry analysis during 3 weeks of culture. Among the PGRs tested, 2,4‐dichlorophenoxyacetic acid (2,4‐D) and 4‐amino‐3,5,6‐trichloropicolinic acid caused a drastic reduction in the 4C‐cell proportion in cell cultures with an increased cell proportion of 8C or higher C‐values. In the case of 2,4‐D application, a reduction of cell viability was observed. A decreasing proportion was also observed in the 8C‐cell population accumulated by 2,4‐D treatment, following transfer back to the medium containing the standard PGR composition. These results suggest that the exogenous application of 2,4‐D arrested the cell cycle at G2 phase in the Doritaenopsis cells, and the removal of 2,4‐D might induce further endoreduplication or recover the mitotic cycle of the G2‐arrested cells.