Effects of R-(1-amino-2-phenylethyl)phosphonic acid on glyceollin accumulation and expression of resistance to Phytophthora megasperma f.sp. glycinea in soybean

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Abstract
(R)-(1-Amino-2-phenylethyl)phosphonic acid (R-APEP), an inhibitor of phenylalanine ammonia-lyase (PAL), was applied to the tap root of 42-h-old soybean (Glycine max. (L.) Merrill cv. Harosoy 63) seedlings during inoculation with zoospores of the incompatible race 1 of Phytophthora megasperma f.sp. glycinea (Pmg1) for 2 h and during a subsequent incubation period. In contrast to L-2-aminooxy-3-phenylpropionic acid, R-APEP was not toxic to the zoospores which remained virulent in presence of the inhibitor. A 50% inhibition of PAL activity in vitro was observed with 4.2 μM R-APEP and with 36 μM of the S-enantiomer. When R-APEP at 330 μM was applied for a total of 36 h to the seedlings, resistance against Pmg 1 was abolished. Such seedlings were indistinguishable in appearance from those seedlings which had been inoculated with the compatible race 3 of Pmg. Roots treated with R-APEP at 330 μM showed a reduction of about 47% in glyceollin content when measured 12 h after inoculation, and with 1 mM a 67% reduction. In contrast, treatment with S-APEP (1 mM) caused only a 20% reduction in glyceollin content. As determined by indirect immunofluorescence of fungal hyphae in cryotome cross-sections of roots, the growth pattern of the incompatible race 1 of Pmg changed to that of the compatible race 3 under conditions where R-APEP caused loss of resistance against Pmg 1. The results support the concept of an important role of glyceollin in resistance of soybean against incompatible races of the fungus.