Peanut stem rot caused by Sclerotium rolfsii is a serious soil-borne disease and poses a threat to the peanut production. The antibiotic fungicide tetramycin has a broad antifungal spectrum against multiple pathogens and possess low environmental risks. In current study, a total of 250 isolates collected from Huanghuai peanut-growing region of China (Henan, Shandong and Hebei Province) were used to establish the baseline sensitivity of S. rolfsii to tetramycin. The baseline sensitivity curve was unimodal and distributed from 0.01 to 0.36 mg/L, with a mean EC₅₀ (50% effective concentration) value of 0.11 ± 0.06 mg/L. Tetramycin also had strong inhibitory activity on the formation and germination of sclerotia. There was no significant correlation of S. rolfsii sensitivity to tetramycin and other commonly used SDHI (succinate dehydrogenase inhibitor), QoI (quinone outside respiration inhibitor) and DMI (demethylation inhibitor) fungicides. Moreover, tetramycin significantly increased the cell membrane permeability and reduced the oxalate acid content. Greenhouse experiments showed that tetramycin has both protective and curative efficacy against S. rolfsii, while protective efficacy was higher than curative efficacy. Anyhow, the bioactivity of tetramycin is similar (curative efficacy) or higher (protective efficacy) than the control fungicide validamycin. In terms of application method, root drench may be more suitable for tetramycin than spraying, because root drench of tetramycin obtained a higher efficacy. These results indicated that tetramycin may be a potential alternative fungicide for the efficient control of peanut stem rot.

Among succinate dehydrogenase inhibitors, only boscalid has been registered in China for controlling gray mold. In Shandong Province of China, it has been more than a decade since the first use of boscalid to control gray mold. In the current study, we monitored the resistance development process of Botrytis cinerea to boscalid, identified the mutation types that occurred in boscalid-resistant isolates, and proposed an original application technique to delay resistance development. A total of 720 B. cinerea isolates collected from tomato and cucumber in Shandong Province from 2014 to 2019 were determined to be sensitive to boscalid. The results showed that the sensitivity of the B. cinerea isolates to boscalid declined gradually over time, with a mean half maximal effective concentration of 0.3 ± 0.02 mg/liter in 2014 and 6.39 ± 1.66 mg/liter in 2019. The proportion of resistant isolates quickly increased from 0.81% in 2014 to 28.97% in 2019. Mutations of P225F, N230I, H272Y, and H272R in the SdhB subunit were responsible for boscalid resistance. Four concurrent mutations (G85A, I93V, M158V, and V168I) in the SdhC subunit were first discovered in Shandong Province, but they did not affect the level of boscalid resistance. Interestingly, this study found that the fruit dipping application, a precise topical application technique, could delay the development of boscalid resistance. Therefore, this application technique provides a new method for resistance management of B. cinerea.

Tetramycin is a new biopesticide that combines high-level and broad-spectrum fungicidal activity, low toxicity, and environmental safety. In this study, 90 Phytophthora capsici isolates obtained from various regions in southern China were characterized for their baseline sensitivity to tetramycin. The protective and curative activities of tetramycin against P. capsici were determined on leaves of pepper, and the control efficacy of tetramycin in greenhouse experiments was also determined. Compared with mycelial growth, the formation of sporangia and the discharge of zoospores were inhibited by lower concentrations of tetramycin, approximately 5 µg ml⁻¹ on V8 media. The frequency distribution curves for the tetramycin sensitivity were unimodal, with mean values for the fungicide concentration that reduced mycelial growth, sporangia formation, and zoospore discharge by 50% compared with the control of 1.18 ± 0.91, 0.64 ± 0.42, and 0.63 ± 0.30 µg ml⁻¹, respectively. In addition, no correlation was observed between tetramycin and other fungicides tested, including mandipropamid, azoxystrobin, mefenoxam, fluazinam, fluopicolide, and famoxadone. Tetramycin exhibited both protective and curative effects against P. capsici in vitro, and its protective activity was better than its curative activity. In greenhouse experiments, tetramycin concentration of 60 and 90 µg ml⁻¹ provided a protective control efficacy of 47.1 to 56.4% and curative efficacy of 43.3 to 52.7%. These results demonstrated that tetramycin could serve as an excellent alternative fungicide to control Phytophthora blight of pepper.

Gray mold caused by Botrytis cinerea is a fungal disease that critically threatens agricultural production, and carbendazim was the first fungicide used to control B. cinerea. However, B. cinerea developed serious resistance to carbendazim, and this fungicide has thus rarely been used in the past decade in China. Due to the extended discontinuation of carbendazim use, the evolution of the resistance of B. cinerea to carbendazim in recent years is unclear, and whether carbendazim can effectively control gray mold is largely unknown. Therefore, this study determined the sensitivity of 407 B. cinerea isolates collected from 2014 to 2018 to carbendazim and the ability of carbendazim to control gray mold in the field. The results showed that the frequency of B. cinerea isolates resistant to carbendazim remained above 95%. Three different mutation types responsible for the resistance of B. cinerea to carbendazim were identified at codon 198 in the β-tubulin gene sequence: E198V (changed from GAG to GTG), E198A (changed from GAG to GCG), and E198K (changed from GAG to AAG). Over the last 5 years, E198V was the major mutation. However, an analysis of its evolution revealed that the percentage of the E198V mutation declined after 2017 to 56.5% in 2018. In addition, the proportion of isolates with the E198K mutation decreased over time, and no isolates with this mutation were found in either 2017 or 2018. The proportion of the E198A mutation increased over the 5-year test period to reach 43.5% in 2018. Furthermore, three greenhouse experiments demonstrated that carbendazim has lost its ability to control gray mold. We attribute the above findings to our results showing that the carbendazim-resistant isolates had no fitness penalties compared with the carbendazim-sensitive isolates for sporulation and mycelial growth. In particular, the E198A mutant isolates exhibited a strong ability to sporulate, suggesting that the E198A mutation might become dominant in the future. Interestingly, the results showed that carbendazim-sensitive isolates could be easily controlled by four conventional fungicides, namely boscalid, procymidone, iprodione, and pyrimethanil, with mean EC₅₀ values of 0.71 ± 0.2 mg liter⁻¹, 1.33 ± 0.39 mg liter⁻¹, 0.59 ± 0.33 mg liter⁻¹, and 6.02 ± 3.02 mg liter⁻¹, respectively. In conclusion, carbendazim has lost its application value and is ineffective for the control of gray mold.

Benzothiazole is a microbial volatile compound with strong antifungal activity against the phytopathogenic fungus Botrytis cinerea, but its mode of action against fungi remains largely unknown. Und...