Feeding Behavior of Riptortus pedestris (Fabricius) on Soybean: Electrical Penetration Graph Analysis and Histological Investigations

SIMPLE SUMMARY: Riptortus pedestris (Fabricius) (Hemiptera: Alydidae) is a major soybean pest with the peak population occurrence during the seed maturity stage from pod filling to harvest. Soybean pods and/or seeds are required for R. pedestris development. However, the feeding strategies employed by this stink bug to feed on soybean are still not clear. In the present study, we recorded the feeding behaviors of R. pedestris on soybean using electropenetrography (EPG). The biological meaning of each waveform was confirmed by histological examination of plant tissues containing stylets or salivary sheath. In total, five phases of waveforms were identified: non-probing, pathway (Rp1), xylem sap ingestion (Rp2), salivation and ingestion (Rp3), and interruption (Rp4). Xylem ingestion (Rp2) was observed during R. pedestris feeding on soybean leaflets, stems, and pods, demonstrating that the stink bug was ingesting xylem sap from vascular tissue. Cell rupture (salivation/ingestion, Rp3) was only recorded during R. pedestris feeding on cotyledon and pods. Histological images showed that stylet tips and the salivary sheath were positioned in the tissues of cotyledon and pods. Taken together, our results demonstrate that R. pedestris uses a cell-rupture strategy to acquire nutrients from soybean pods and/or seeds while utilizing salivary sheath tactics to obtain water from xylem sap. These findings provide insightful information to understand the interactions between R. pedestris and the soybean plant. ABSTRACT: Riptortus pedestris (Fabricius) is a major agricultural pest feeding on soybean pods and seeds. The large populations occur during seed maturity stages from pod filling to harvest. Its infestation results in shriveled and dimpled seeds while vegetative structures (leaflet and stem) remain green, known as “Stay Green” syndrome. Additional evidence also demonstrates that soybean pods and seeds are required for Riptortus pedestris development. However, the feeding behavior strategies employed by this stink bug to feed on soybean plants are still not clear. In the present study, the feeding behaviors of R. pedestris on soybean plants were recorded by electropenetrography (EPG), and a waveform library was created for this species. A total of five phases of waveforms—nonprobing, pathway (Rp1), xylem sap ingestion (Rp2), salivation and ingestion (Rp3), and interruption (Rp4)—were identified. Non-probing waveforms Z and NP and pathway (Rp1) were found in all tested plant structures (leaflet, stem, cotyledon, and pods). Waveform Rp2 (xylem sap ingestion, xylem ingestion) was primarily recorded during R. pedestris feeding on leaflets and stems, while Rp3 (salivation/ingestion) was only observed during feeding on cotyledon and pods. Histological examinations confirmed that correlation between Rp2 and stylet tip positioning in the xylem vessel in leaflets and stems. Stylet tips end in the tissues of cotyledon and pods when Rp3 is recorded. Taken together, our results demonstrate that R. pedestris ingests xylem sap from vegetative tissues of soybean (leaflet and stem) via a salivary sheath strategy to obtain water. It mainly acquires nutrients from soybean pods and/or seeds using cell-rupture tactics. This study provided insightful information to understand the field occurrence patterns of “Stay Green” syndrome, which may have important implications for pest control.