Inhibitory Effects of Stilbenes on the Growth of Three Soybean Pathogens in Culture

Abstract

The effects of resveratrol and pterostilbene on in vitro growth of three soybean pathogens were tested to determine whether these stilbenic compounds could potentially be targets to increase innate resistance in transgenic soybean plants. Growth of Macrophomina phaseolina, Rhizoctonia solani, and Sclerotinia sclerotiorum was measured on solid and in liquid media amended with resveratrol and pterostilbene (concentration in the media of resveratrol at 100 μg/ml and pterostilbene at 25 μg/ml). All three fungi were very sensitive to pterostilbene in potato dextrose agar (PDA), which reduced colony area of each of the three pathogens to less than half of the control 3 days after incubation. The three fungal pathogens were less sensitive to resveratrol compared with pterostilbene; however, area under the curve (AUC) calculated from colony areas measured over 3 days was significantly (P < 0.05) less than the control for S. sclerotiorum and R. solani on PDA with resveratrol or pterostilbene. AUC for M. phaseolina on PDA with pterostilbene was significantly (P < 0.05) lower than the control whereas, on PDA with resveratrol, AUC for M. phaseolina was lower than the control but the difference was nonsignificant (P > 0.05). AUC for all three fungi was significantly lower (P < 0.05) on PDA with pterostilbene than with resveratrol. In potato dextrose broth (PDB) shake cultures, AUC for all three fungi was significantly (P < 0.01) lower in pterostilbene than in the control. AUC for R. solani and S. sclerotiorum was significantly lower (P < 0.01) in resveratrol than the control, whereas AUC for M. phaseolina in resveratrol was lower, but not significantly (P > 0.05) different from the control. AUC in pterostilbene was highly significantly (P < 0.01) lower than in resveratrol for M. phaseolina and significantly (P < 0.05) lower for R. solani but the difference for S. sclerotiorum was nonsignificant (P > 0.05). There was a trend for lower mass accumulation of all three fungi in either pterostilbene or resveratrol compared with the control during the course of the experiment; however, S. sclerotiorum appeared to recover from the effects of pterostilbene between days 2 and 4. Results of biochemical analyses of the PDB over time indicated that the three fungi degraded resveratrol, with nearly 75% reduction in concentration in M. phaseolina, 80% in S. sclerotiorum, and 60% in R. solani PDB cultures by day 4 of fungal growth. M. phaseolina and S. sclerotiorum were able to resume growth after early inhibition by resveratrol after its concentration was reduced in the cultures through degradation, whereas R. solani was less efficient in resveratrol degradation and was not able to overcome its inhibitory effects on growth. The capacity to degrade pterostilbene was lowest in M. phaseolina compared with S. sclerotiorum and R. solani and the recovery of M. phaseolina cultures after initial growth inhibition by pterostilbene was minimal. The potential products of resveratrol and pterostilbene degradation by fungi were identified to be dimers and various oxidation products.