Characterization of Fusarium graminearum Isolates Resistant to Both Carbendazim and a New Fungicide JS399-19

Fusarium head blight (FHB) of wheat and other cereals, caused mainly by Fusarium graminearum, is one of the most economically important diseases worldwide, especially in the United States and China. The benzimidazole fungicides, particularly carbendazim (MBC), have been consistently used during the period of wheat heading and flowering in areas with warm and moist weather to control FHB in China for over 30 years. The effectiveness of MBC, however, has been threatened by the emergence of resistant pathogen populations in the field. JS399-19 (experimental number; a.i. 2-cyano-3-amino-3-phenylancryic acetate) is a novel cyanoacrylate fungicide discovered and patented by the Jiangsu Branch of National Pesticide Research & Development South Center of China. To evaluate the potential risk of resistance development in MBC-resistant F. graminearum isolates to this new fungicide JS399-19, five isolates each of MBC-resistant or -sensitive, which were classified into three different sensitivity phenotypes, such as sensitive (S), moderately resistant (MR), and highly resistant (HR) to MBC, were selected to induce JS399-19-resistant mutants by selecting resistance on potato sucrose agar (PSA) plates amended with JS399-19 at 10 μg/ml. In this way, a total of 24 JS399-19-resistant mutants were obtained from all tested MBC-resistant or -sensitive isolates. All 50 single-spore progenies of each of the resistant mutants could grow normally on PSA plates amended with JS399-19 at 10 μg/ml, indicating stability of resistance to this fungicide. Also, all of the resistant mutants maintained their resistance to JS399-19 and/or MBC through eight transfers on PSA plates for 40 days and when stored on PSA slants at 4°C for 60 days. The mycelial growth and conidial production capacity were decreased in 52.4% of the resistant mutants, indicating that a fitness cost was associated with JS399-19-resistant phenotypes of F. graminearum isolates. However, most of the mutants resistant to both MBC and JS399-19 exhibited high sexual reproduction capacity and pathogenicity as their parental isolates. Nevertheless, the majority of these mutants possessed fitness levels comparable to their parents. The results on the efficacy of the two fungicides for controlling FHB incited by the fungicide-resistant mutants were generally consistent with those of the in vitro sensitivity tests. JS399-19 was effective in controlling FHB caused by MBC-resistant isolates under field conditions, while it was not effective in controlling FHB caused by isolates resistant to JS399-19 or those that were resistant to both MBC and JS399-19. Moreover, the efficacy of the mixture of MBC and JS399-19 was also significantly lower in controlling FHB caused by the isolates resistant to both MBC and JS399-19 than the efficacy against the disease caused by the sensitive isolates, the MBC-resistant isolates, or the JS399-19-resistant isolates. The results suggest that JS399-19 possessed a high risk in development of resistance in MBC-resistant and -sensitive F. graminearum isolates, and this double resistance to both of these fungicides could presumable emerge and create a major problem since both these fungicides are extensively used in China. Therefore, careful use of JS399-19 should be followed to delay resistance development in natural populations of F. graminearum, avoid unexpected control failures, and sustain the usefulness of MBC and the new product JS399-19.