The A2′N Mutation of the RDL γ-Aminobutyric Acid Receptor Conferring Fipronil Resistance in Laodelphax striatellus (Hemiptera: Delphacidae)

Kawase, Ayumi; Kinoshita, Ayako; Abe, Reiko; Hama, Masako; Kawahara, Nobuyuki; Hirase, Kangetsu

doi:10.1603/ec10391

Cited by 70 publications

(67 citation statements)

References 37 publications

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“…SF-RDL genes To co-express wild-type and mutant SF-RDL genes, the plasmid pENTR-pA-pAc 21) was used. The plasmid pENTR1A-Sf-RDL was digested with BamHI and Not I, and the BamHINot I fragment containing the wild-type SF-RDL gene was inserted into the BamHI-Not I site of pENTR-pA-pAc; the resulting plasmid was designated as pENTR pA-pAc-Sf-RDL.…”

Section: Construction Of Vectors To Co-express Wild-type and Mutantmentioning

confidence: 99%

“…[16][17][18][19] In a previous study, the A2′N mutation was associated with fipronil resistance and was suggested to be a heterozygous mutation in S. furcifera 20) and L. striatellus. 21) A membrane potential assay showed that the A2′N mutation in the L. striatellus RDL GABA receptor subunit (LS-RDL) confers fipronil resistance in the heterozygous expression of wild-type and A2′N mutant LS-RDL genes. 21) In this study, we used heterozygous expression and heterozygous receptors to indicate GABA receptors expressed on co-transfected cells with an equal amount of wild-type and mutant genes.…”

Section: Introductionmentioning

confidence: 99%

“…21) A membrane potential assay showed that the A2′N mutation in the L. striatellus RDL GABA receptor subunit (LS-RDL) confers fipronil resistance in the heterozygous expression of wild-type and A2′N mutant LS-RDL genes. 21) In this study, we used heterozygous expression and heterozygous receptors to indicate GABA receptors expressed on co-transfected cells with an equal amount of wild-type and mutant genes. This notation was used to facilitate the description of the GABA receptors expressed on cotransfected cells, and no genetic mechanism or specific assembly patterns were implied.…”

Section: Introductionmentioning

confidence: 99%

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Fipronil resistance in Sogatella furcifera: Molecular cloning and functional expression of wild-type and mutant RDL GABA receptor subunits

et al. 2012

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The whitebacked planthopper Sogatella furcifera is a serious pest of rice that has developed fipronil resistance in Asia. We found a novel R340Q mutation in the cytoplasmic loop between M3 and M4 of the S. furcifera RDL γ-aminobutyric acid (GABA) receptor subunit (SF-RDL). The R340Q mutation was found in clones carrying the A2′N mutation, which was previously implicated in fipronil resistance and suggested to be a heterozygous mutation. To investigate the influence of mutations, Drosophila Mel-2 cells were transfected with wild-type or mutant SF-RDL genes, either individually or together. A membrane potential assay showed that the influence of the A2′N · R340Q double mutation on fipronil resistance was more profound than that of the A2′N single mutation in the heterozygous expression of wild-type and mutant SF-RDL genes.

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Section: Construction Of Vectors To Co-express Wild-type and Mutantmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Fipronil resistance in Sogatella furcifera: Molecular cloning and functional expression of wild-type and mutant RDL GABA receptor subunits

et al. 2012

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“…Ser 301 provides low resistance to the phenylpyrazole fipronil that does not impact the use of this insecticide in the field (9,43,46,47). However, fipronil-resistant strains isolated from two species of planthopper have been shown to contain an Ala to Asn, rather than Ser or Gly, mutation at the equivalent 301 site, found only in the heterozygous state (48,49). The lack of homozygous Asn individuals was proposed to be a result of lethality, supported by the reduced GABA median effective concentration (EC 50 ) observed in electrophysiological data (49).…”

mentioning

confidence: 99%

“…However, fipronil-resistant strains isolated from two species of planthopper have been shown to contain an Ala to Asn, rather than Ser or Gly, mutation at the equivalent 301 site, found only in the heterozygous state (48,49). The lack of homozygous Asn individuals was proposed to be a result of lethality, supported by the reduced GABA median effective concentration (EC 50 ) observed in electrophysiological data (49). Mutations resulting in lethality can only be viable in a heterozygous state, or more permanently, in a duplication of the type described in this study.…”

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confidence: 99%

Gene duplication in the major insecticide target site,Rdl, inDrosophila melanogaster

Remnant

Good

Schmidt

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The Resistance to Dieldrin gene, Rdl, encodes a GABA-gated chloride channel subunit that is targeted by cyclodiene and phenylpyrazole insecticides. The gene was first characterized in Drosophila melanogaster by genetic mapping of resistance to the cyclodiene dieldrin. The 4,000-fold resistance observed was due to a single amino acid replacement, Ala 301 to Ser. The equivalent change was subsequently identified in Rdl orthologs of a large range of resistant insect species. Here, we report identification of a duplication at the Rdl locus in D. melanogaster. The 113-kb duplication contains one WT copy of Rdl and a second copy with two point mutations: an Ala 301 to Ser resistance mutation and Met 360 to Ile replacement. Individuals with this duplication exhibit intermediate dieldrin resistance compared with single copy Ser 301 homozygotes, reduced temperature sensitivity, and altered RNA editing associated with the resistant allele. Ectopic recombination between Roo transposable elements is involved in generating this genomic rearrangement. The duplication phenotypes were confirmed by construction of a transgenic, artificial duplication integrating the 55.7-kb Rdl locus with a Ser 301 change into an Ala 301 background. Gene duplications can contribute significantly to the evolution of insecticide resistance, most commonly by increasing the amount of gene product produced. Here however, duplication of the Rdl target site creates permanent heterozygosity, providing unique potential for adaptive mutations to accrue in one copy, without abolishing the endogenous role of an essential gene.T he single point mutation in the Resistance to dieldrin (Rdl) gene represents one of the most significant cases of target site resistance to an insecticide yet observed. Cyclodiene resistance was reported in 62% of insecticide resistant species in the 1980s, following widespread use of cyclodiene insecticides, including dieldrin, which started in the 1950s (1). The nature of the genetic target, Rdl, was discovered after dieldrin was discontinued because of the widespread evolution of resistance in many species. Rdl was first discovered in Drosophila melanogaster using a positional cloning approach. High homology to human GABA receptors confirmed it was the first insect ligand-gated chloride channel subunit identified (2-4). A point mutation in the chloride channel pore-lining domain, replacing alanine 301 with serine, was present in all resistant D. melanogaster strains (5). This mutation provided 4,000-fold resistance when homozygous and lower levels of resistance in heterozygotes (2, 6). The homologous mutation was subsequently found in a large number of cyclodiene-resistant species from many insect orders (7-9), as well as a glycine replacement at the homologous site in some resistant strains of Drosophila simulans and other species (5, 10, 11).Characterization of deficiency lines and inversions in D. melanogaster showed that Rdl is an essential gene (3). Thus, the Ala 301 to Ser or Gly mutation in Rdl exhibits unique properties, p...

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KNOCKDOWN OF THE IONOTROPIC γ‐AMINOBUTYRIC ACID RECEPTOR (GABAR) RDL GENE DECREASES FIPRONIL SUSCEPTIBILITY OF THE SMALL BROWN PLANTHOPPER, Laodelphax striatellus (HEMIPTERA: DELPHACIDAE)

Niu

et al. 2015

Arch Insect Biochem Physiol

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Insect γ-aminobutyric acid receptors (GABARs) are important molecular targets of cyclodiene and phenylpyrazole insecticides. Previously GABARs encoding rdl (resistant to dieldrin) genes responsible for dieldrin and fipronil resistance were identified in various economically important insect pests. In this study, we cloned the open reading frame cDNA sequence of rdl gene from fipronil-susceptible and fipronil-resistant strains of Laodelphax striatellus (Lsrdl). Sequence analysis confirmed the presence of a previously identified resistance-conferring mutation. Different alternative splicing variants of Lsrdl were noted. Injection of dsLsrdl reduced the mRNA abundance of Lsrdl by 27-82%, and greatly decreased fipronil-induced mortality of individuals from both susceptible and resistant strains. These data indicate that Lsrdl encodes a functional RDL subunit that mediates susceptibility to fipronil. Additionally, temporal and spatial expression analysis showed that Lsrdl was expressed at higher levels in eggs, fifth-instar nymphs, and female adults than in third-instar and fourth-instar nymphs. Lsrdl was predominantly expressed in the heads of 2-day-old female adults. All these results provide useful background knowledge for better understanding of fipronil resistance related ionotropic GABA receptor rdl gene expressed variants and potential functional differences in insects.

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The A2′N Mutation of the RDL γ-Aminobutyric Acid Receptor Conferring Fipronil Resistance in Laodelphax striatellus (Hemiptera: Delphacidae)

Cited by 70 publications

References 37 publications

Fipronil resistance in Sogatella furcifera: Molecular cloning and functional expression of wild-type and mutant RDL GABA receptor subunits

Fipronil resistance in Sogatella furcifera: Molecular cloning and functional expression of wild-type and mutant RDL GABA receptor subunits

Gene duplication in the major insecticide target site,Rdl, inDrosophila melanogaster

KNOCKDOWN OF THE IONOTROPIC γ‐AMINOBUTYRIC ACID RECEPTOR (GABAR) RDL GENE DECREASES FIPRONIL SUSCEPTIBILITY OF THE SMALL BROWN PLANTHOPPER, Laodelphax striatellus (HEMIPTERA: DELPHACIDAE)

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