Flubendiamide resistance and Bi-PASA detection of ryanodine receptor G4946E mutation in the diamondback moth (Plutella xylostella L.)

• Moderate and high level resistance to flubendiamide was identified.
• A point mutation causing flubendiamide resistance was investigated.
• Different length fragments were detected in SS, RR, and RS genotypes.
• A correlation between mutation gene frequencies and resistance ratios.
• The highest resistant ratio and gene mutation frequency were in ZC population.

The extensive application of flubendiamide has led to increasingly prominent development of resistance in diamondback moth, Plutella xylostella. Here we report that the moderate and high level resistance to flubendiamide was identified in a laboratory-selected and two field-collected strains of P. xylostella. The resistance ratios were tested in the lab-selected resistant strains (R), and two field strains (BY and ZC). Compared with the S strain, the R strain showed extended larval development time, decreased pupation rate, emergencing rate, and male adult longevity. The realized heritability (h2 = 0.135) implies the high risk of flubendiamide resistance development in P. xylostella. A Bi-PASA (bi-directional PCR amplification of specific allele)-based method was successfully developed to detect the point mutation (G4946E) potentially causing flubendiamide resistance in diamondback moth, in which different fragments 866 bp + 340 bp, 866 bp + 568 bp, and 866 bp + 568 bp + 340 bp were presented in SS, RR and RS stains, respectively. The predominant genotype was 83.33% SS homozygote in the S strain, 80.77% RR homozygote in ZC population, and 73.08% RS heterozygote in BY population, respectively. Current results showed the significant correlation between the frequencies of the allele carrying G4946E mutation (51.92%, 55.77% and 90.38% for R, BY and ZC, respectively) and the resistance ratios (40.72, 24.24 and 1779.24-folds for R, BY and ZC, respectively) in the three strains/populations. In addition, the relative PxRyR mRNA transcript level in the R strain was 2.938 ± 0.53 folds as compared with the S strain (1.0-fold).

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