title: Frequency detection of imidacloprid resistance allele in Aphis gossypii field populations by real-time PCR amplification of specific-allele (rtPASA).
abstract: The Aphis gossypii Glover (Hemiptera: Aphididae) is one of the most serious pests worldwide, and imidacloprid has been widely used to control this insect pest. Just like other classes of insecticides, the resistance to imidacloprid has been found in A. gossypii. An amino acid mutation (R81T) in the nicotinic acetylcholine receptor (nAChR) beta1 subunit was detected in the imidacloprid-resistant A. gossypii collected from Langfang (LF) and Dezhou (DZ) cities. To estimate the R81T mutation frequency of A. gossypii field populations, a simple, rapid and accurate rtPASA (real-time PCR amplification of specific allele) protocol was developed. The performance of the rtPASA protocol was evaluated by comparing with the data generated by a cPASA (competitive PCR amplification of specific allele) method from 50 individual genotypes. The R81T allele frequencies of the LF population (34.7%±1.3%) and DZ population (45.2%±5.2%) estimated by the rtPASA protocol matched the frequencies (LF 38.1%, DZ 48.2%) deduced by the cPASA method in specimens. The results indicated that the rtPASA format was applicable for the detection of mutation associated with imidacloprid resistance and will allow rapid and efficient monitoring of A. gossypii resistance in field populations in a high throughput format.
authors.affiliation: Key Laboratory of Integrated Pest Management in Crops Ministry of Agriculture, Institute of Plant Protection Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Integrated Pest Management in Crops Ministry of Agriculture, Institute of Plant Protection Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Li
authors.initials: L
authors.lastname: Cui
authors.affiliation: Key Laboratory of Integrated Pest Management in Crops Ministry of Agriculture, Institute of Plant Protection Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Xibao
authors.initials: X
authors.lastname: Xu
authors.affiliation: Key Laboratory of Integrated Pest Management in Crops Ministry of Agriculture, Institute of Plant Protection Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: chrui@ippcaas.cn.
authors.forename: Changhui
authors.initials: C
authors.lastname: Rui
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001042:Aphids / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D005787:Gene Frequency; D003368:Gossypium / Q000254:growth & development / Q000469:parasitology*; D007093:Imidazoles / Q000494:pharmacology*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D000073943:Neonicotinoids; D009574:Nitro Compounds / Q000494:pharmacology*; D010935:Plant Diseases / Q000469:parasitology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism
chemical_list: D007093:Imidazoles; D019476:Insect Proteins; D007306:Insecticides; D000073943:Neonicotinoids; D009574:Nitro Compounds; D011978:Receptors, Nicotinic; C082359:imidacloprid
keywords: Allele frequency; Aphis gossypii; Imidacloprid resistance; Real-time PASA (rtPASA); cPASA