Autoclaved and filter sterilized liquid media in maize anther culture: significance of activated charcoal |
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Authors: | B Büter S M Pescitelli K Berger J E Schmid P Stamp |
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Institution: | (1) Institute of Plant Sciences, Swiss Federal Institute of Technology (ETH), Eschikon 33, CH-8315 Lindau, Switzerland;(2) Biotechnology and Plant Genetics Dept., DowElanco, P.O. Box 68955, Indianapolis, IN, USA;(3) Present address: Institute of Pharmacy, University of Basel, Kleinhüninger Anlage 3, CH-4057 Basel, Switzerland |
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Abstract: | Summary Medium sterilization techniques (autoclaving, filter sterilization and separate sterilization of medium components), combined
with preculture exposure to activated charcoal (AC) were evaluated for effects on maize anther culture response. The addition
of AC to filter sterilized medium had no effect on the number of embryo-like-structures (ES) produced. For autoclaved medium,
pre-culture AC treatment resulted in a 3-fold increase in ES yield over medium lacking AC. When AC was included, autoclaved
medium was more productive than filter sterilized medium. Autoclaved media without AC gave lower response than filter sterilized
medium. Separate sterilization of sucrose or FeEDTA was beneficial for media autoclaved in the absence of AC. However, when
all components were autoclaved together in the presence of AC, there was no advantage to separate sterilization. The maximum
ES frequency (224.6 ES/100 anthers) was obtained with the genotype ETH-M 52 cultured in autoclaved medium which had been exposed
to AC (5 g/L) for 96 h prior to culture initiation. It is supposed that the higher ES frequencies observed with AC-treated,
autoclaved media were due to the availability of glucose and fructose following heat-induced hydrolysis of sucrose and the
AC-mediated adsorption of inhibitory compounds produced during autoclaving. |
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Keywords: | Maize Anther culture Autoclaving Medium toxicity Activated charcoal |
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