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Published in Crop Sci 10:479-481 (1970)
© 1970 Crop Science Society of America
677 S. Segoe Rd., Madison, WI 53711 USA
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Rapid Detection of {sigma}-Hydroxycinnamic Acid and Beta-Glucosidase in Melilotus alba1

F. A. Haskins and H. J. Gorz2

The tests utilize small samples of leaf tissue crushed on sheets of filter paper. For detection of {sigma}-hydroxycinnamic acid ({sigma}-HCA), a crude preparation containing sweetclover ß-glucosidase is added to a spot of crushed tissue to insure hydrolysis of glucosidically bound {sigma}-HCA. An extract containing cis-o-HCA glucoside is added to another tissue spot to serve as the substrate for the detection of ß-glucosidase activity. Finally, all spots are moistened with NaOH and are scored for1 fluorescence under ultraviolet light. Results of qualitative and quantitative tests on plants representing various genotypes are compared. The importance of exposing plants to sunlight before testing for {sigma}-HCA is discussed. The tests are particularly useful in genetic studies, in the development and maintenance of breeding lines, and in testing for contamination in low-{sigma}-HCA sweetclover varieties.

Key Words: Sweetclover • Coumarin

1 Contribution from the Crops Research Division, Agricultural Research Service, U. S. Department of Agriculture, and the Nebraska Agricultural Experiment Station, Lincoln, Neb. Supported in part by the National Science Foundation (Grant Nos. CB-1148 and GB-8280). Published with the approval of the Director as Paper No. 2720, Journal Series, Nebraska Agr. Exp. Sta.

2 Bert Rodgcrs Professor of Agronomy, University of Nebraska, and Research Geneticist, Crops Research Division, ARS, USDA, Lincoln, Neb. The technical assistance of Mrs. Linda Helton, David Gilchrist, and Ralph Kulm is gratefully acknowledged.

Received for publication December 12, 1969.




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