Glyceollin I in soybean-cyst nematode interactions : spatial and temporal distribution in roots of resistant and susceptible soybeans

Abstract

Accumulation of the phytoalexin glyceollin I in roots of soybean (Glycine max [L.] Merr.) following inoculation with race 1 of Heterodera glycines Ichinohe, the soybean cyst nematode (SCN), was determined in a whole-root system by high performance liquid chromatography (HPLC) and in a cross-section system by a radioimmunoassay procedure. In the whole-root system, roots were harvested from controls and nematode-inoculated seedlings immediately after inoculation and at 2-day intervals for 8 days. The roots were extracted with ethanol, and the extracts were subjected to HPLC. Glyceollin I was not detected in roots of either resistant cultivar Centennial or susceptible cultivar Ransom immediately after inoculation with SCN but steadily accumulated in large quantity in roots of Centennial. Accumulation of glyceollin I in roots of Ransom following nematode inoculation was minimal. In the cross-section system, 3-day-old soybean seedlings were inoculated with juvenile nematodes, and root segments containing a single nematode were dissected from inoculated plants at 4-hour intervals under a dissecting microscope. The root segments were embedded in ice and cut into 16-micrometer sections with a cryostat microtome. The spatial and temporal distribution of glyceollin I was determined with a radioimmunoassay procedure specific for the phytoalexin. Glyceollin I was found to accumulate in tissues immediately adjacent to the head region of the nematode in Centennial but not in Ransom. Glyceollin I was detected 8 hours after nematode penetration, and the concentration increased steadily up to 0.3 micromole per milliliter in Centennial 24 hours after penetration.