Resistance risk assessment for fludioxonil in Bipolaris maydis

Abstract

Bipolaris maydis (anamorph: Cochliobolus heterostrophus) is the causal agent of Southern Corn Leaf Blight (SCLB), leading to huge annually losses worldwide. Although fludioxonil, a phenylpyrrole fungicide with a broad spectrum of activity, was introduced in the 1990s, no baseline sensitivity has been established for B. maydis. One hundred field isolates were used to establish a baseline sensitivity of B. maydis against fludioxonil during 2015–2016. The results showed that the baseline sensitivity was distributed as a unimodal curve with a mean EC50 value of 0.044±0.022μgmL−1. With repeated exposure to fludioxonil, a total of five fludioxonil-resistant mutants (RF>100, RF=Resistance factor) were obtained in the laboratory. Compared with the parental isolates, the five fludioxonil-resistant mutants showed decreased fitness in sporulation and virulence, and exhibited different features of sensitivity to various stresses (oxidation and osmotic pressure, cell membrane and cell wall inhibitors), but not in mycelial growth on PDA without stress amendation. The five fludioxonil-resistant mutants showed a positive cross-resistance between fludioxonil and the dicarboximide fungicide procymidone, but not between fludioxonil and boscalid or fluazinam. All mutants exhibited stable resistance to fludioxonil after 10 transfers, as indicated by resistance factor values that ranged from 116.82 to 445.59. When treated with 1.0 M NaCl, all the fludioxonil-resistant mutants showed greater mycelial glycerol content than corresponding parental isolates. Sequencing alignment results of Bmos1 indicated that mutant R27-5 had a single point mutation (Z1125K), while the mutant R104 had a 34-bp deletion fragment between the codons of amino acid residues 1125 to 1236 and encodes a putative attenuated 1133-AA protein. The 34-bp deletion fragment led to not only a 11-AA deletion(DNAVNQKLAVR), but also the resulting frameshift mutation and early stop. The mutations of R27-5 and R104 were located in the Rec domain of the Bmos1 gene. No mutations at the Bmos1 were detected in the other three resistant mutants R27-1, R27-2 and R32.