Gold-conjugated arabinogalactan-protein and other lectins as ultrastructural probes for the wheat/stem rust complex.
Arabinogalactan-protein (AGP, "beta-lectin") was isolated from leek seeds, tested for specificity, conjugated with gold colloids, and used as a cytochemical probe to detect beta-linked bound sugars in ultrathin sections of wheat leaves infected with a compatible race of stem rust fungus. Similar sections were probed with other gold-labeled lectins to detect specific sugars. AGP-gold detected beta-glycosyl in all fungal walls and in the extrahaustorial matrix. Other lectin gold conjugates localized galactose in all fungal walls except in walls of the haustorial body. Limulus polyphemus lectin bound only to the outermost layer of intercellular hyphal walls of the fungus. Binding of these lectins was inhibited by their appropriate haptens and was diminished or abolished in specimens pretreated with protease, indicating that the target substances in the tissue were proteinaceous or that polysaccharides possessing affinity to the lectin probes had been removed by the enzyme from a proteinaceous matrix by passive escape. Binding of Lotus tetragonolobus lectin was limited to the two outermost fungal wall layers but was not hapten-inhibitable. Limax flavus lectin, specific for sialic acids, had no affinity to any structure in the sections. In the fungus, the most complex structure was the outermost wall layer of intercellular hyphal cells; it had affinity to all lectins tried so far, except to Limax flavus lectin and to wheat germ lectin included in an earlier study. In the host, AGP and the galactose-specific lectins bound to the inner domain of the wall in areas not in contact with the fungus. At host cell penetration sites, affinity to these lectins often extended throughout the host wall, confirming that it is modified at these sites. Pre-treatment with protease had no effect on lectin binding to the host wall. After protease treatment, host starch granules retained affinity to galactose-specific lectins, but lost affinity for AGP.
Rohringer
R
R
Research Station, Agriculture Canada, Winnipeg, Manitoba.
Chong
J
J
Gillespie
R
R
Harder
D E
DE
D050260:Carbohydrate Metabolism; D005658:Fungi / Q000378:metabolism* / Q000648:ultrastructure; D005685:Galactans / Q000378:metabolism*; D005779:Immunodiffusion; D007150:Immunohistochemistry; D037102:Lectins / Q000378:metabolism*; D011135:Polysaccharides, Bacterial; D011506:Proteins / Q000378:metabolism*
D005685:Galactans; D037102:Lectins; D011135:Polysaccharides, Bacterial; D011506:Proteins; C005653:arabinogalactan