Summary
Two solid pectic media were devised for mutually exclusive detection of extracellular polygalacturonase and pectin lyase produced by fungi including the vascular parasite of tomato Verticillium albo-atrum. These media allowed detection of pectinase-defective mutants. Polygalacturonase detection medium contains non-methylated polygalacturonan (sodium polypectate) is buffered at pH 5.0 (Na citrate, 0.05 M) and is calcium-free. In contrast pectin lyase medium contains polymethylgalacturonan (pectin), is buffered at pH 8.0 (HEPES, 0.05 M) and contains calciumrich agar. When glucose was added to the media for selection of catabolite-resistant mutants, enzyme synthesis was still evident, whereas in comparable conditions in liquid culture production was almost completely repressed. This apparent discrepancy is discussed in terms of the influence of basal synthesis, colony biomass and accumulation of oligouronides which repress induced synthesis and activity.
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Abbreviations
- CR:
-
catabolite repression
- CTAB:
-
cetyltrimethyl ammoniumbromide
- GALA:
-
galacturonic acid
- NAPP:
-
sodium polypectate
- PG:
-
polygalacturonase
- PL:
-
pectin lyase
- TBA:
-
thiobarbituric acid
- UGALA:
-
unsaturated galacturonic acid
References
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Durrands, P.K., Cooper, R.M. Development and analysis of pectic screening media for use in the detection of pectinase mutants. Appl Microbiol Biotechnol 28, 463–467 (1988). https://doi.org/10.1007/BF00268215
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DOI: https://doi.org/10.1007/BF00268215