Pectolytic enzyme detection media
From Bugwoodwiki
Purpose
Ingredients
| Ingredient | 300 ml |
|---|---|
| Cold DH2O | 300 ml |
| Polygalacturonic A | 3 g |
| Yeast extract | 3 g |
| cresol red solution | 3 ml |
| agar | 4.5 g |
Instructions
- Pour cold dH2O in 1 L beaker.
- Slowly add polygalacturonic A--be careful not to clump.
- Add remaining ingredients in same way.
- Heat to nearly boiling while stirring continuously.
- Adjust to pH 7.3 with 1 N NaOH.
- Autoclave 15 minutes at 15 psi and 120 °C.
- Bring pH to 7.8 with 1 N NaOH (turns pink), then cool.
- Grow cultures 48 hours.
- Flood with 2 N HCl or 1% aqueous solution hexadecyl-trimethyl ammonium bromide
- Clear zones indicate presence of pectolytic enzymes.
Notes
References
- Cuppels, D. and Kelman, A. 1974. Evaluation of selective media for isolation of soft-rot bacteria from soil and plant tissue. Phytopathology 64: 468-475; modified by Y. B. Di and A. Kelman, and subsequently by M.C.M. Perombelon and E. M. Burnett, unpublished data.
Contributed by
From the Virginia Polytechnic Institute and State University Mediabook; Orignially created by Robert Wick; contributed by Mary Ann Hansen.
