In the present study, we demonstrated the use and optimization of the tetra-primer ARMS-PCR procedure to detect and analyze the frequency of the @entity70151 and @entity70152 mutations in resistant field populations of CPB. The @entity70151 mutation was detected in 72%, 84%, 52% and 64% of Bahar, Dehpiaz, Aliabad and Yengijeh populations, respectively. Overall frequencies of the @entity70152 mutation were 12%, 8% and 16% of Bahar, Aliabad and Yengijeh populations, respectively. No @entity70152 point mutation was found among samples from Dehpiaz field population. Moreover, only 31% and 2% of samples from the resistant field populations were homozygous for @entity70151 and @entity70152 mutations, respectively. No individual simultaneously had both @entity70152 and @entity70153 / @entity70151 point mutations. The incidence of individuals with both @entity70153 and @entity70151 point mutations in the samples from Bahar, Dehpiaz, Aliabad, and Yengijeh populations were 31.5%, 44.7%, 41.6%, and 27.3% respectively. Genotypes determined by the tetra-primer ARMS-PCR method were consistent with those determined by PCR sequencing. There was no significant correlation between the mutation frequencies and resistance levels in the resistant populations, indicating that other mutations may contribute to this variation. Polymorphism in the partial L. decemlineata cDNA @entity10810 gene Ldace2 of four field populations was identified by direct sequencing of PCR-amplified fragments. Among 45 novel mutations detected in this study, @entity70154 mutation was found across all four field populations that likely contribute to the @entity10810 insensitivity. Site-directed mutagenesis and protein expression experiments are needed for a more complete evaluation.

Target site insensitivity mutations in the XXXX enzyme confer resistance to organophosphorous insecticides in Leptinotarsa decemlineata (Say).

@entity70153 :: ('p|SUB|S|291|G', 'ProteinMutation') :: ['S291G'] @entity70152 :: ('p|SUB|I|392|T', 'ProteinMutation') :: ['I392T'] @entity70151 :: ('p|SUB|R|30|K', 'ProteinMutation') :: ['R30K'] @entity10810 :: ('43', 'Gene') :: ['AChE'] @entity70154 :: ('p|SUB|T|29|P', 'ProteinMutation') :: ['T29P']
@entity10810 :: (43,Gene) :: ['AChE']
