Increasing ploidy level in cell suspension cultures of Doritaenopsis by exogenous application of 2,4-dichlorophenoxyacetic acid.
Increasing ploidy level in cell suspension cultures of Doritaenopsis by exogenous application of 2,4-dichlorophenoxyacetic acid.
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通过外源应用 2,4-二氯苯氧基乙酸提高 Doritaenopsis 细胞悬浮培养物的倍性水平。
DOI：
10.1034/j.1399-3054.2001.1120119.x
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发表时间：
2001
影响因子：
6.4
通讯作者：
M. Mii
中科院分区：
文献类型：
--
作者：
K. Mishiba;Tomonori Okamoto;M. Mii
为了阐明植物细胞培养中倍性变异的原因，我们试图通过改变培养基中的植物生长调节剂（PGR）来改变Doritaenopsis四倍体品种细胞培养中的倍性分布。将原悬浮培养的细胞（其已维持在含有0.1mg l-1 1-萘乙酸和1 mg l-1苄基腺嘌呤的培养基中）转移到各种结冷胶固化培养基上，单次施用PGR，并在培养3周期间使用流式细胞术分析来检查细胞的倍性分布。在所测试的PGR中，2，4-二氯苯氧基乙酸（2，4-D）和4-氨基-3，5，6-三氯吡啶甲酸导致细胞培养物中4C细胞比例急剧降低，8 C或更高C值的细胞比例增加。在2，4-D应用的情况下，观察到细胞活力降低。在转移回含有标准PCR组合物的培养基后，在通过2，4-D处理积累的8 C-细胞群体中也观察到降低的比例。这些结果表明，外源性2，4-D使Doritaenopsis细胞的细胞周期阻滞在G2期，去除2，4-D可能会进一步诱导胞内复制或使阻滞在G2期的细胞恢复有丝分裂周期。
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.