Co-occurrence of subunit B and C mutations in respiratory complex II confers high resistance levels to pyflubumide and cyenopyrafen in the two-spotted spider mite Tetranychus urticae (Acari: Tetranychidae)

[BACKGROUND] Pyflubumide and cyenopyrafen are respiratory complex II (complex II) inhibitors. Previous quantitative trait locus analyses suggested associations of I260V and S56L in complex II subunit B (B-I260V) and subunit C (C-S56L) with pyflubumide and cyenopyrafen resistance, respectively, in Tetranychus urticae. However, although resistant strains had been selected separately by these acaricides, all strains were homozygous for both B-I260V and C-S56L. Hence, the effects of each mutation on resistance development remain unclear. [RESULTS] We established strains homozygous for B-I260V with C-S56 (B-I260V_I260V/C-S56_S56) and for C-S56L with B-I260 (B-I260_I260/C-S56L_S56L). High resistance levels (LC₅₀ > 1000 mg L⁻¹) to pyflubumide and cyenopyrafen was not conferred by B-I260V or C-S56L alone. Next, we prepared intermixed strains by crossing B-I260V_I260V/C-S56_S56 and B-I260_I260/C-S56L_S56L. Selection of the intermixed strains by either acaricide caused very high resistance levels (LC₅₀ ≥ 10 000 mg L⁻¹) to both acaricides and fixed both mutations. Allele-selected recoupling of the mutations without acaricide selection also conferred very high resistance levels to both acaricides in the intermixed strains. Unlike these, B-I260V or C-S56L alone conferred very high and high resistance levels to cyflumetofen, respectively. [CONCLUSION] We conclude that the effect of individual mutations characteristically varies among complex II inhibitors. Moreover, very high resistance levels to pyflubumide and cyenopyrafen is conferred by the co-occurrence of B-I260V and C-S56L mutations, which alone have limited effects on resistance level