Detection of Citrus Leaf and Seed Glycoproteins using Biotinylated Lectin Probes

in HortScience
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  • 1 Horticultural Research Laboratory, U.S. Department of Agriculture-Agricultural Research Service, 2120 Camden Road, Orlando, FL 32803

leaf and seed glycoproteins were detected after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotting onto polyvinylidene difluoride (PVDF) membrane when probed with biotinylated lectin at 1 μg·ml-1. Four lectins representing three carbohydrate-binding groups were used as probes. A preformed avidin-biotin-complex (ABC) was used to detect the glycoprotein-bound lectins and resulted in dark bands and little background staining. Concanavalin A (ConA) and wheat germ agglutinin (WGA) resulted in the darkest-staining bands. The four Citrus spp. and one related species studied had unique seed glycoprotein profiles when probed with ConA and WGA. This procedure might be useful in clarifying citrus taxonomy, providing genetic markers, and in physiological studies involving glycoproteins.

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