Author
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Reed, Barbara |
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De Noma, Jeanine |
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Submitted to: Society for In Vitro Biology Proceedings
Publication Type: Abstract Only Publication Acceptance Date: 3/17/2015 Publication Date: 6/17/2015 Citation: Reed, B.M., De Noma, J.S. 2015. Practical aspects of running DOE for improving growth media for in vitro plants. Society for In Vitro Biology Proceedings. Vol 51: S21-22. Interpretive Summary: Experiments using statistical design software to improve plant tissue culture growth medium are complicated and require complex setups. Once the experimental design is set and the treatments calculated, media sheets and mixing charts must be developed. This talk will include the types of details needed to set up these complex experiments. These include caculation tables, stock solutions, charts indicating how much of each stock is to be used for each treatment and any factors that are part of the basal medium (agar, vitamins and growth regulators). These experiments are designed so that all of the 20 to 45 treatments can be run at one time or they can be run sequentially in smaller, more manageable sets. Logistical planning also includes the number of growth containers needed and the space required in the growth room. Generally test-medium preparation requires one or two days and experiment setup two to three days for each passage. Useful data may include a quality rating, shoot length and multiplication or callus size and fresh/dry weight, leaf or stem color, size or abnormalities and biochemical analyses. Photographs of the shoots or callus at the end of the experiment are useful as a comparison to graphical data and rating scales. Large amounts of data are generated and these can be managed on a spreadsheet. Technical Abstract: Experiments using DOE software to improve plant tissue culture growth medium are complicated and require complex setups. Once the experimental design is set and the treatment points calculated, media sheets and mixing charts must be developed. Since these experiments require three passages on the same treatment before data is taken, it is very important that the calculations are correct and the procedures clear so they can be repeated accurately. It may be necessary to develop new stock solutions unique to the experiment. Calculating the amount of each stock solution needed for a treatment is most easily done with a spreadsheet program. The third step is a chart indicating how much of each stock is to be used for the required number of containers of each treatment and any factors that are part of the basal medium (agar, vitamins and growth regulators). All treatments can be run at one time or sequentially in smaller more manageable sets. In addition, one experiment can require 60-80 containers per genotype tested and considerable space in the growth room. Generally test-medium preparation requires one or two days and planting two to three days for each passage. Useful data may include a quality rating, shoot length and multiplication or callus size and fresh/dry weight, leaf or stem color, size or abnormalities and biochemical analyses. Photographs of the shoots or callus at the end of the experiment are useful as a comparison to graphical data and rating scales. Large amounts of data are generated and these can be managed on a spreadsheet. |
