Novel mutations and mutation combinations of ryanodine receptor in a chlorantraniliprole resistant population of Plutella xylostella (L.)

• Published in: Scientific reports Vol. 4; no. 1; p. 6924
• Main Authors: Guo, Lei, Liang, Pei, Zhou, Xuguo, Gao, Xiwu
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.11.2014; Nature Publishing Group
• Subjects: 45/70; 45/77; 631/208/737; 631/337; Alleles; Amino Acid Sequence; Amino Acid Substitution; Animals; Conserved Sequence; Female; Frequency analysis; Gene Expression; Glutamic acid; Glutamine; Glycine; Humanities and Social Sciences; Insect Proteins - genetics; Insecticide Resistance - genetics; Insecticides; Isoleucine; Leucine; Male; Methionine; Molecular Sequence Data; Moths - genetics; multidisciplinary; Mutation; Mutation Rate; ortho-Aminobenzoates; Pesticide resistance; Polymerase chain reaction; Polymerase Chain Reaction - methods; Population studies; Ryanodine Receptor Calcium Release Channel - genetics; Ryanodine receptors; Science; Valine
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep06924

A previous study documented a glycine to glutamic acid mutation (G4946E) in ryanodine receptor (RyR) was highly correlated to diamide insecticide resistance in field populations of Plutella xylostella (Lepidoptera: Plutellidae). In this study, a field population collected in Yunnan province, China, exhibited a 2128-fold resistance to chlorantraniliprole. Sequence comparison between resistant and susceptible P. xylostella revealed three novel mutations including a glutamic acid to valine substitution (E1338D), a glutamine to leucine substitution (Q4594L) and an isoleucine to methionine substitution (I4790M) in highly conserved regions of RyR. Frequency analysis of all four mutations in this field population showed that the three new mutations showed a high frequency of 100%, while the G4946E had a frequency of 20%. Furthermore, the florescent ligand binding assay revealed that the RyR containing multiple mutations displayed a significantly lower affinity to the chlorantraniliprole. The combined results suggested that the co-existence of different combinations of the four mutations was involved in the chlorantraniliprole resistance. An allele-specific PCR based method was developed for the diagnosis of the four mutations in the field populations of P. xylostella .