Detection of ryanodine receptor G4911E and I4754M mutation sites and analysis of binding modes of diamide insecticides with RyR on Galeruca daurica (Coleoptera: Chrysomelidae)

Abstract

In recent years, the leaf beetle Galeruca daurica has broken out in the northern grasslands of Inner Mongolia, its management still mainly depends on chemical control using traditional insecticides or with novel action. The study was aim to identify mutation locus associated with resistance to diamide insecticides in field population of G. daurica, to provide a reference for rational selection of insecticides and to avoid the rapid resistance development to diamide insecticides. We cloned the full length of the ryanodine receptor gene of G. daurica (GdRyR), constructed 3D model and transmembrane regions by homologous modeling based on deduced amino acid sequence. Two potential mutation loci (Gly4911Glu and Ile4754Met) and allelic mutation frequencies were detected in individuals of G. daurica. In addition, their binding patterns to two diamide insecticides (chlorantraniliprole, cyantraniliprole) were analyzed separately using a molecular docking method. The full-length cDNA sequence of GdRyR (GenBank accession number: OP828593) was obtained by splicing and assembling, which is 15,399 bp in length and encodes 5,133 amino acids. The amino acid similarity of GdRyR with that of other Coleopteran insects were 86.70%–91.33%, which possessed the typical structural characteristics. An individual resistance allelic mutation frequency test on fifty field leaf beetles has identified 12% and 32% heterozygous individuals at two potential mutation loci Gly4911Glu and Ile4754Met, respectively. The affinity of the I4754M mutant model of GdRyR for chlorantraniliprole and cyantraniliprole was not significantly different from that of the wild type, and all had non-covalent interactions such as hydrogen bonding, hydrophobic interactions and π-cation interactions. However, the G4911E mutant model showed reduced affinity and reduced mode of action with two diamide insecticides, thus affecting the binding stability of the ryanodine receptor to the diamide insecticides. In conclusion, the G4911E mutation in GdRyR may be a potential mechanism for the development of resistance to diamide insecticides on G. daurica and should be a key concern for resistance risk assessment and reasonable applications of diamide insecticides for control in future. Moreover, this study could provide a reference for ryanodine receptor structure-based insecticides design.

Sublethal effects of chlorantraniliprole on biological characteristics, detoxifying enzyme activity and gene expression profile in the Allium mongolicum Regel leaf beetle Galeruca daurica (Coleoptera: Chrysomelidae)

“…In recent years, there has been an outbreak of the leaf beetle G. daurica in the grasslands of Inner Mongolia, which has severely restricted the sustainable development of the livestock and grass farming, and chemical control has been the main means of control relied upon. Chlorantraniliprole, as new O‐amidobenzamide chemical structure, is a highly effective, broad‐spectrum, low‐toxicity insecticide with a stomach‐toxic effect, which has good efficacy against Coleoptera and Lepidoptera, which is also recommended for controlling G. daurica (Ren et al., 2022; Zhang et al., 2022). In the present study, chlorantraniliprole had an inhibitory effect on its growth and development of the 3rd instar larvae of G. daurica , and the inhibitory effect was significant with increasing concentration, indicating that the larvae were highly susceptible to chlorantraniliprole and could exert good control effects.…”

3D-QSAR Combination with Molecular Dynamics Simulations to Effectively Design the Active Ryanodine Receptor Agonists against Spodoptera frugiperda

“…Since RyRs of different insect species are highly sequence conserved and homologous, the two mutation sites of the S. frugiperda RyR (G4891E and I4734M) were obtained by a sequence alignment with the two mutation sites of the Plutella xylostella RyR (G4946E and I4790M), the Spodoptera exigua RyR (G4900E and I4743M) and the Chilo suppressalis RyR (G4910E and I4758M). − The software chimera 1.16 was used to construct models of the S. frugiperda RyR mutant G4891E (mutating GLY to GLU) and mutant I4734M (mutating ILE to MET).…”