Unknown,Transcriptomics,Genomics,Proteomics

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Anopheles funestus amidst a South/North shift in the role of key resistance genes across Malawi

ABSTRACT: Resistance to pyrethroids, the only insecticide approved for bednets, threatens control of the major malaria vector, Anopheles funestus, in Malawi. To improve the management of such resistance countrywide, it is crucial to understand the dynamics and mechanisms driving resistance in the field. In this study the levels of insecticide resistance were determined across the highly endemic densely populated lake and southern agricultural area. Insecticide resistance to pyrethoids was assessed using standardized WHO bioassay methods and resistant mosquitoes were hybridized to susceptible mosquitoes. This microarray analysis revealed the key role of cytochrome P450 genes such as CYP6P9a, CYP6P9b and CYP6M7. However, a significant shift in the over-expression of these CYP450s was detected across a south/north transect, with CYP6M7 more highly over-transcribed in the two northern collection sites and the tandemly duplicated genes, CYP6P9a and CYP6P9b, more greatly over-transcribed in the south.

ORGANISM(S): Anopheles funestus

SUBMITTER: Kayla Barnes

PROVIDER: E-MTAB-3342 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The aim of this experiment was to compare the transciptome of the fall armyworm (Spodoptera frugiperda) strain SUS (a laboratory insecticide-susceptible standard) with organophosphate (OP) and pyrethroid (PYR) resistant strains were collected in cornfields located in Minas Gerais and Mato Grosso States, Brazil, in 2008 and maintained in the laboratory under selection with chlorpyrifos (at increasing discriminating doses from 100 up to 400 M-BM-5g of insecticide/g of insect in a topical bioassay) or lambda-cyhalothrin (from 8.4 up to 27 M-BM-5g of insecticide/g of insect) respectively The custom microarray used in this study was designed using the Agilent eArray platform (Agilent Technologies). A SurePrint HD (8M-CM-^W15k) expression array was designed using the base composition and the best probe methodologies to design sense orientation 60-mer probes with a 3M-bM-^@M-2 bias. The FAW EST database (SPODOBASE) was used as the reference transcriptome (Negre, Hotelier et al. 2006). These sequences are derived from 8 cDNA libraries as follows: Sf1F: Fat body, Sf1H: Hemocyte, Sf1M: Midgut, Sf1P: Pool of various tissues, Sf2H: Immune Challenged hemocytes, Sf2L: Sf21 Cell lines sequences, Sf2M: Xenobiotic Induced Midgut and Sf9L: Sf9 cell lines sequences. All assembled contigs and singlets were kindly sent by the website maintainers. The BLAST2GO software v.2.3.1 (http://www.blast2go.org) was used to annotate the EST database. 60-mer probes were designed for all 7,552 assembled contigs and 5,519 annotated singlets (BlastX), totaling 13,071 sequences. For contigs encoding detoxification enzymes (P450s, GSTs and CEs) three probes were designed. Additional probe groups for 15 control genes were also included. For reference all sequences are included in the zip file with array data. References: Negre, V., T. Hotelier, et al. (2006). "SPODOBASE : an EST database for the lepidopteran crop pest Spodoptera." BMC Bioinformatics 7: 322. Two-condition experiment. Slide 1: SUS vs. OP S. fugiperda strains. Slide 2: SUS vs. PYR S. fugiperda strains. Biological replicates: 4 pools of RNA extracted from four pools of 5 second instar larvae. Technical Replicates: None, the biological replicates incorporated a dye swap. Total replication: four replicates for each strain.