Extensive gene duplication of acetylcholinesterase associated with organophosphate resistance in the two‐spotted spider mite

Monocrotophos‐resistant two‐spotted spider mites (TSSMs), Tetranychus urticae, are known to possess three mutations on the acetylcholinesterase (AChE) gene (Tuace) that are involved in target site insensitivity. Cross‐strain comparison of three strains (highly resistant AD, moderately resistant PyriF and susceptible UD strains) revealed that resistant strains have relatively more Tuace copies than the UD strain and that the levels of transcript were directly proportional to copy numbers. AChEs from the AD and PyriF strains had similar Vmax values to those of AChE from the UD strain but increased Km and reduced kcat constants, suggesting that the mutated, resistant form of AChE may carry a fitness cost. Relative copy numbers of Tuace in field populations varied from 2.4 to 6.1, correlating well with their levels of resistance (r2= 0.895). These results are suggestive of the involvement of Tuace gene duplication in resistance. Thus, monocrotophos resistance in TSSMs appears to have evolved through a combination of mutation accumulation and extensive gene duplication.

[1]  D. Kwon,et al.  Cloning of a sodium channel gene and identification of mutations putatively associated with fenpropathrin resistance in Tetranychus urticae , 2010 .

[2]  D. Kwon,et al.  Acetylcholinesterase point mutations putatively associated with monocrotophos resistance in the two-spotted spider mite , 2010 .

[3]  T. Lenormand,et al.  Forty Years of Erratic Insecticide Resistance Evolution in the Mosquito Culex pipiens , 2007, PLoS genetics.

[4]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[5]  T. Lenormand,et al.  Independent duplications of the acetylcholinesterase gene conferring insecticide resistance in the mosquito Culex pipiens. , 2007, Molecular biology and evolution.

[6]  Ju Il Kim,et al.  Identification and characterization of ace1-type acetylcholinesterase likely associated with organophosphate resistance in Plutella xylostella , 2005 .

[7]  B. Croft,et al.  Ecological and genetic factors influencing evolution of pesticide resistance in tetranychid and phytoseiid mites , 1988, Experimental & Applied Acarology.

[8]  W. Helle Genetics of resistance to organophosphorus compounds and its relation to diapause inTetranychus urticae Koch (Acari) , 1962, Tijdschrift Over Plantenziekten.

[9]  R. Veitia Gene dosage balance: deletions, duplications and dominance. , 2005, Trends in genetics : TIG.

[10]  C. Malcolm,et al.  The unique mutation in ace‐1 giving high insecticide resistance is easily detectable in mosquito vectors , 2004, Insect molecular biology.

[11]  T. Nabeshima,et al.  An amino acid substitution attributable to insecticide-insensitivity of acetylcholinesterase in a Japanese encephalitis vector mosquito, Culex tritaeniorhynchus. , 2004, Biochemical and biophysical research communications.

[12]  D. Fournier,et al.  Acetylcholinesterase alterations reveal the fitness cost of mutations conferring insecticide resistance , 2004, BMC Evolutionary Biology.

[13]  Yoshio Anazawa,et al.  Sequence of a cDNA encoding acetylcholinesterase from susceptible and resistant two-spotted spider mite, Tetranychus urticae. , 2003, Insect biochemistry and molecular biology.

[14]  M. Navajas,et al.  Mechanisms of resistance to organophosphates in Tetranychus urticae (Acari: Tetranychidae) from Greece. , 2002, Insect biochemistry and molecular biology.

[15]  D. Fournier,et al.  Levels of Total Acetylcholinesterase in Drosophila melanogaster in Relation to Insecticide Resistance , 2001 .

[16]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[17]  M. Houck Mites: Ecological and Evolutionary Analyses of Life-History Patterns , 1993 .

[18]  S. C. Lee,et al.  Toxicological test methods and AChE inhibition of organophosphorus acaricides of two spotted spider mite, Tetranychus urticae , 1992 .

[19]  M. H. Tageldin A rapid detection of organophosphorus resistance with insensitive acetylcholinesterase in spider mites Tetranychus urticae Koch on cotton , 1990 .

[20]  Si-Woo Lee,et al.  Method Comparison of Chemical-Resistance Level Determination and Field Resistance of Two-Spotted Spider Mite, Tetranychus urticae Koch to Benzomate, Cyhexatin, and Dicofol , 1986 .

[21]  Á. Lagunes-Tejeda,et al.  The occurrence of resistance to pesticides in arthropods , 1981 .

[22]  C. Shute,et al.  The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscope. , 1966, Journal of cell science.

[23]  H. R. Smissaert Cholinesterase Inhibition in Spider Mites Susceptible and Resistant to Organophosphate , 1964, Science.

[24]  K. Courtney,et al.  A new and rapid colorimetric determination of acetylcholinesterase activity. , 1961, Biochemical pharmacology.