In vivo identification of Bacillus thuringiensis Cry4Ba toxin receptors by RNA interference knockdown of glycosylphosphatidylinositol-linked aminopeptidase N transcripts in Aedes aegypti larvae

Saengwiman, Suchada; Aroonkesorn, Aratee; Dedvisitsakul, Plaipol; Sakdee, Somsri; Leetachewa, Somphob; Angsuthanasombat, Chanan; Pootanakit, Kusol

doi:10.1016/j.bbrc.2011.03.085

Cited by 34 publications

(33 citation statements)

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“…aegypti α-amylase AAEL010540 (48% identity with aamy1) could bind Cry toxins. No cadherin or APN that were previously associated with Cry resistance in insects [34–37] or showed to bind Bti Cry toxins in mosquitoes [38–40] were differentially transcribed in the LiTOX strain, suggesting that if they are involved in Bti resistance in LiTOX, this is not related to an altered expression but rather to changes in protein sequence affecting their affinity for Cry toxins. Indeed, one cadherin, eight APNs and four α-amylases displayed differential SNPs leading to non-synonymous changes between the resistant and susceptible strains.…”

Section: Discussionmentioning

confidence: 99%

“…aegypti larvae, 12 contained differential SNPs, and non-synonymous differential changes affected the sequence of 8 of them (Table 2). In the APN2 AAEL005808 recently shown to be a functional receptor of Cry4Ba toxin [40], 4 non-synonymous changes were nearly fixed in the LiTOX strain (both phenotypes), suggesting ongoing selective sweep on this gene. One non-synonymous change affected the sequence of the APN1 AAEL012778 previously shown to bind Cry11Aa [46].…”

Section: Discussionmentioning

confidence: 99%

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Gene expression patterns and sequence polymorphisms associated with mosquito resistance to Bacillus thuringiensis israelensis toxins

et al. 2014

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BackgroundDespite the intensive use of Bacillus thuringiensis israelensis (Bti) toxins for mosquito control, little is known about the long term effect of exposure to this cocktail of toxins on target mosquito populations. In contrast to the many cases of resistance to Bacillus thuringiensis Cry toxins observed in other insects, there is no evidence so far for Bti resistance evolution in field mosquito populations. High fitness costs measured in a Bti selected mosquito laboratory strain suggest that evolving resistance to Bti is costly. The aim of the present study was to identify transcription level and polymorphism variations associated with resistance to Bti toxins in the dengue vector Aedes aegypti. We used RNA sequencing (RNA-seq) for comparing a laboratory-selected strain showing elevated resistance to Bti toxins and its parental non-selected susceptible strain. As the resistant strain displayed two marked larval development phenotypes (slow and normal), each phenotype was analyzed separately in order to evidence potential links between resistance mechanisms and mosquito life-history traits.ResultsA total of 12,458 genes were detected of which 844 were differentially transcribed between the resistant and susceptible strains. Polymorphism analysis revealed a total of 68,541 SNPs of which 12,571 SNPs exhibited more than 40% frequency difference between the resistant and susceptible strains, affecting 2,953 genes. Bti resistance is associated with changes in the transcription level of enzymes involved in detoxification and chitin metabolism. Among previously described Bti-toxin receptors, four alkaline phosphatases (ALPs) were differentially transcribed between resistant and susceptible larvae, and non-synonymous changes affected the protein sequence of one cadherin, six aminopeptidases (APNs) and four α-amylases. Other putative Cry receptors located in lipid rafts, such as flotillin and glycoside hydrolases, were under-transcribed and/or contained non-synonymous substitutions. Finally, immunity-related genes showed contrasted transcription and polymorphisms patterns between the two developmental resistant phenotypes, suggesting the existence of trade-offs between Bti-resistance, life-history traits and immunity.ConclusionsThe present study is the first to analyze the whole transcriptome of Bti-resistant mosquitoes by RNA-seq, shedding light on the importance of studying both transcription levels and sequence polymorphism variations to get a comprehensive view of insecticide resistance.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-926) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Gene expression patterns and sequence polymorphisms associated with mosquito resistance to Bacillus thuringiensis israelensis toxins

et al. 2014

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“…The intensive use of Bacillus thuringiensis has led to the development of resistance in natural mosquito population (Boyer et al, 2012). However, some reports suggest a cross development against Bt, due to the increase in detoxifying activity of enzyme in xenobiotic reaction, decreasing activity of mid gut protease and a modification of specific receptor of mid gut (Jurat-Fuentes et al, 2004;Boyer et al, 2007;Saengwiman et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Synergistic effect of certain insecticides combined with Bacillus thuringiensis on mosquito larvae

Narkhede

Suryawanshi

Koli

et al. 2017

J Entomol Acarol Res

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For effective vector control it is essential to formulate new preparations having multiple action against the vector pest. Developing combined formulation of biopesticide and chemical pesticide is one of the novel concept to fight against the vectors with new weapons; however, compatibility of biopesticide i.e. Bacillus thuringiensis (Bt) and chemical pesticide is a real hurdle. In this investigation, local isolate Bacillus thuringiensis SV2 (BtSV2) was tested for its compatibility with various available mosquito larvicides. Temephos was most compatible with BtSV2 than with other tested pesticides. These two compatible agents were tested for larvicidal potential. Our study revealed that the synergistic effect of both agents reduces LC50 value by 30.68 and 22.36% against the Ae. aegypti and An. stephensi, respectively. The larvicidal potential increased when compared to individual pesticides. It was also observed a biochemical change in larvae after the TBT (Temephos + Bacillus thuringiensis) combination treatment; it involves decreased level of alpha esterase, acetylcholine esterase and protein while level of beta esterase and acid phosphatase was unchanged and alkaline phosphatase activity was increased. Increased potential of combined formulation may be due to altered physiological condition.

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“…Numerous studies have proposed multiple Cry toxin receptors that mediate toxicity; these include cadherin-like proteins, glycosylphosphatidylinositol (GPI)-anchored aminopeptidase N (APN), GPI-anchored alkaline phosphatases (ALPs), and glycolipids (for a review, see reference 18). For instance, in our earlier studies, two different membrane-bound proteins, i.e., GPI-linked ALP (GPI-ALP) and GPI-linked APN (GPI-APN), have been identified as putative receptors mediating toxicity for the Cry4Ba toxin in the Aedes aegypti larvae (7,19). Of particular interest, ALPs (EC 3.1.3.1; phosphomonoester hydrolases), ubiquitous enzymes widely found in microorganisms and animal kingdoms, are a highly conserved family of largely distributed cell surface and secreted metalloenzymes; many are GPI-anchored glycoproteins (for a review, see reference 13).…”

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confidence: 99%

Aedes aegypti Membrane-Bound Alkaline Phosphatase Expressed in Escherichia coli Retains High-Affinity Binding for Bacillus thuringiensis Cry4Ba Toxin

Thammasittirong

Dechklar

Leetachewa

et al. 2011

Appl Environ Microbiol

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Glycosylphosphatidylinositol-linked alkaline phosphatase (GPI-ALP) from the epithelial membrane of the larval midgut of Aedes aegypti was previously identified as a functional receptor of the Bacillus thuringiensis Cry4Ba toxin. Here, heterologous expression in Escherichia coli of the cloned ALP, lacking the secretion signal and GPI attachment sequences, and assessment of its binding characteristics were further investigated. The 54-kDa His tag-fused ALP overexpressed as an inclusion body was soluble when phosphate buffer (pH 7.5) was supplemented with 8 M urea. After renaturation in a nickel-nitrilotriacetic acid (Ni-NTA) affinity column, the refolded ALP protein was able to retain its phosphatase activity. This refolded ALP also showed binding to the 65-kDa activated Cry4Ba toxin under nondenaturing (dot blot) conditions. Quantitative binding analysis using a quartz crystal microbalance revealed that the purified ALP immobilized on a gold electrode was bound by the Cry4Ba toxin in a stoichiometry of approximately 1:2 and with high affinity (dissociation constant [K d ] of ϳ14 nM) which is comparable to that calculated from kinetic parameters (dissociation rate constant [k off ]/binding constant [k on ]). Altogether, the data presented here of the E. coli-expressed ALP from A. aegypti retaining high-affinity toxin binding support our notion that glycosylation of this receptor is not required for binding to its counterpart toxin, Cry4Ba.

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In vivo identification of Bacillus thuringiensis Cry4Ba toxin receptors by RNA interference knockdown of glycosylphosphatidylinositol-linked aminopeptidase N transcripts in Aedes aegypti larvae

Cited by 34 publications

References 25 publications

Gene expression patterns and sequence polymorphisms associated with mosquito resistance to Bacillus thuringiensis israelensis toxins

Gene expression patterns and sequence polymorphisms associated with mosquito resistance to Bacillus thuringiensis israelensis toxins

Synergistic effect of certain insecticides combined with Bacillus thuringiensis on mosquito larvae

Aedes aegypti Membrane-Bound Alkaline Phosphatase Expressed in Escherichia coli Retains High-Affinity Binding for Bacillus thuringiensis Cry4Ba Toxin

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