Avoidance of precipitation and carbohydrate breakdown in autoclaved plant tissue culture media.
The extent of breakdown of fructose and glucose derived from sucrose in the medium of Murashige and Skoog (1962) during autoclaving was investigated by polarographic measurement. Although not present in the original MS medium but often used in place of FeSO4 + Na2-EDTA, FeNa-EDTA was found to be primarily responsible for catalyzing the breakdown of these monosaccharides. It would therefore be good practice to autoclave FeNa-EDTA separate from the carbohydrate constituents of the medium in order to reduce the formation of toxic substances derived from the latter's breakdown. Autoclaving FeNa-EDTA separately has the additional advantage of preventing precipitation of certain micronutrient elements. Further precipitation can be avoided by autoclaving FeNa-EDTA and KH2PO4 together, but separately, from other components of the medium. By eliminating precipitation and minimizing the breakdown of monosaccharides during autoclaving, it is possible to improve the quality of the medium without resorting to sterilization by filtering.
Research and Development, Hilleshög AB, Box 302, S-261 23, Landskrona, Sweden.
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Schenk
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Hsiao
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Bornman