The Initiation and Characterization of Suspension Cell Cultures of Tradescantia Clones 03 and 4430 and Their Use in The Activation of Promutagens (Phenylenediamine; Ethylene Dibromide)
Anderson, Van Allen
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https://hdl.handle.net/2142/71758
Description
Title
The Initiation and Characterization of Suspension Cell Cultures of Tradescantia Clones 03 and 4430 and Their Use in The Activation of Promutagens (Phenylenediamine; Ethylene Dibromide)
Author(s)
Anderson, Van Allen
Issue Date
1987
Department of Study
Genetics and Development
Discipline
Genetics
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Agriculture, Plant Culture
Abstract
Plant assays utilizing Tradescantia clone 4430 and T. paludosa clone 03 are the most widely used plant assays in the study of environmental mutagenesis. Liquid suspension cell cultures of these clones were established and were used to investigate the activation of m -phenylenediamine, ethylene dibromide and the 5A(E) fraction of 4-nitro- o -phenylenediamine. The cell lines were characterized as to their medium requirements, growth cycle, cell weight, doubling time of cells, protein content per cell and pH changes of the medium. The ability of these plant cell lines to activate specific promutagens was determined using the plant cell/microbe coincubation assay. This work demonstrated that a careful determination of the treatment parameters must be completed and determined methodological improvements for the plant cell/microbe coincubation assay. These treatment parameters included the concentration range of the test compound, the concentration of plant cells, the coincubation period, and the growth stage of the plant cells in the reaction mixture. The optimum treatment parameters for the activation of mPDA, a model compound for the aniline based herbicides, using clone 03 cells were 2.5 $\mu$moles mPDA/plate, 125 mg/ml mid-log phase clone 03 cells, and a 1 h coincubation period. These optimal conditions produced a two to three-fold increase in the mean number of TA98 revertants per plate over the initial, non-optimized conditions. The optimal conditions for activating mPDA with clone 4430 cells were 2.5 $\mu$moles mPDA/plate, 125 mg/ml mid to late-log phase clone 4430 cells, and a 2 h coincubation period. These optimal conditions produced a four to eight-fold increase in the mean number of TA98 revertants per plate over the initial, non-optimized conditions. The optimal conditions for activating EDB with clone 03 cells were 60 pmoles EDB/plate, 125 mg/ml late-log phase clone 03 cells and a 2.5 h coincubation period. The optimal conditions for activating the 5A(E) fraction with clone 03 cells were 0.2 $\mu$l 5A(E) fraction/plate, 50 mg/ml late-log phase clone 03 cells and a coincubation period of 1.5 h. This study demonstrated the usefulness of Trandescantia cell lines in the assessment of the hazards posed by xenobiotics in the environment to the public health due to the ability of plants to activate these compounds to mutagenic agents.
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