Expression of resistance in bean pods to an incompatible isolate of Pseudomonas syringae

Abstract

Isolates of Pseudomonas syringae from hairy vetch were either virulent (compatible) or avirulent (incompatible) to bean. When bean pods were injected at high inoculum levels (5 × 10⁷ cells/ml), both compatible and incompatible isolates multiplied rapidly and caused water-soaking of injected tissues; populations of both isolates were similar at the end of log phase. Lesions caused by the incompatible isolate turned brown by 3 days after inoculation and did not expand; those caused by the compatible isolate remained water-soaked and expanded into the adjoining, noninoculated tissues. At low inoculum levels (5 × 10⁴ cells/ml), the compatible isolate exhibited the same pattern of multiplication and lesion development as at the high inoculum level. The incompatible isolate, on the other hand, caused necrotic flecking within the injected area by 24 h, and these small lesions remained limited. Appearance of the necrotic flecks was correlated with a progressive decline in bacterial populations; after5 days, the population of incompatible bacteria was about a hundredfold lower than that of compatible ones. Populations of incompatible bacteria within and between the necrotic flecks declined rapidly, indicating the potential accumulation of inhibitors of bacterial growth as a result of an inducible resistance mechanism.The phytoalexin phaseollin reached 280 μg/g fresh weight by 3 days after inoculation of bean pods with incompatible bacteria. However, this compound was not involved in resistance. Neither compatible nor incompatible isolates of the bacterium were significantly inhibited when grown in a medium saturated with phaseollin. When other antibiotic substances in plant extracts were sought, substantial amounts of inhibitory compounds were detected only in the ethanolic extract (fraction soluble in ethyl acetate) of pods previously inoculated with an incompatible isolate of P. syringae. Only low levels of inhibition were obtained from identical extracts of uninfected pods or from pods inoculated with a compatible isolate of P. syringae. Bacterial growth was completely prevented in a bean infusion medium containing 1 g fresh weight equivalent of pod tissue challenged with the incompatible isolate. The active compound(s) was nonfluorescent, had a UV absorption maximum at 286 nm, and gave a positive reaction with reagents for phenolic compounds.