Effects of Fungicides on Fungal Development, Conidiophore Morphology, and Conidial Releases from Single Strawberry Powdery Mildew Colonies Assessed Using an Electrostatic Technique

We evaluated the effects of fungicides on strawberry powdery mildew (PM) fungus, Podosphaera aphanis, using an electrostatic technique. Thirty-six fungicides were sprayed on single colonies of P. aphanis on leaves of strawberry seedlings (Fragaria × ananassa Duchesne ex Rozier). Colony development varied depending on the tested fungicides. Particularly, pyraziflumid, triflumizole, triforine, polyoxin, sodium hydrogen carbonate + copper wettable powder, and flutianil + mepanipyrim were highly effective for reducing colony development. P. aphanis colonies were histochemically stained to observe the morphological characteristics of fungal cells forming normal and abnormal conidiophores. Abnormal conidiophores were classified into seven types based on their morphological and cytological characteristics. Finally, asexual conidia were collected from single P. aphanis colonies on the leaves spray-treated with fungicides using a dielectrically polarized insulator plate (electrostatic spore collector); conidia attracted to the insulator plates were counted using a high-fidelity digital microscope. Most tested fungicides highly inhibited the production and/or germination of asexual conidia. The germination of asexual conidia was observed only in thiophanate-methyl (methyl benzimidazole carbamates fungicides; MBC fungicides) and azoxystrobin (quinone outside inhibitors; QoI fungicides). Assessing with the electrostatic technique, we clarified that P. aphanis has developed resistance to both thiophanate-methyl and azoxystrobin. Thus, the methodological assessment analyzing the colony development and the number of conidia released from single colonies will be helpful information for screening effective fungicides

Effect of (−)-Epigallocatechin Gallate to Staphylococcal Enterotoxin A on Toxin Activity

Staphylococcal enterotoxin A (SEA) functions both as superantigens that stimulate non-specific T cell proliferation as well as potent gastrointestinal toxins. We previously reported that (−)-epigallocatechin gallate (EGCG) binds to SEA. Therefore, the ability of EGCG to inhibit SEA toxin activity was examined. As a result, EGCG significantly decreased SEA-induced expression and production of interferon gamma (IFN-γ). In addition, EGCG inhibited SEA-induced spleen cell proliferation. To investigate the role of the galloyl group in EGCG on SEA cytotoxicity in more detail, the effect of the binding of a hydroxyl group at position 3 of the galloyl group in EGCG to SEA on SEA cytotoxicity was examined using two methylated EGCG. SEA cytotoxicity was significantly controlled in both (−)-3′′-Me-EGCG and (−)-4′′-Me-EGCG. These results suggest that EGCG inhibits toxic activity via direct interaction with SEA or without any interaction with SEA. The binding affinity between SEA and EGCG under in vivo conditions was examined using a model solution. Although after treatment under acidic and alkaline conditions, the presence of protein and the digestive tract model solution, EGCG still interacted with SEA. Our studies are the first to demonstrate the effect of the binding of EGCG to SEA on toxin activity