Tissue-Specific Transcript Profiling for ABC Transporters in the Sequestering Larvae of the Phytophagous Leaf Beetle Chrysomela populi

Strauß, Anja; Wang, Ding; Stock, Magdalena; Gretscher, René R.; Groth, Marco; Boland, Wilhelm; Burse, Antje

doi:10.1371/journal.pone.0098637

Cited by 41 publications

(28 citation statements)

References 142 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2) and the four ABCB p-glycoprotein paralogues in D. melanogaster are all found in the adult midgut and/or Malpighian tubules. A transcriptomic analysis of the leaf beetle Chysomela populii also found a large number of ABC genes, predominately belonging to the B, C and G subfamilies, enriched in the Malpighian tubules and/or guts (Strauss et al, 2014). Two of the four ABC genes most consistently overexpressed in pyrethroid-resistant populations are enriched in the Malpighian tubules (ABCB4: ABAP006363) or Malpighian tubules plus midguts (ABCC12: AGAP007917).…”

Section: Resultsmentioning

confidence: 99%

TheAnopheles gambiaeATP‐binding cassette transporter family: phylogenetic analysis and tissue localization provide clues on function and role in insecticide resistance

Pignatelli

Ingham

Balabanidou

et al. 2017

Insect Molecular Biology

View full text Add to dashboard Cite

The role of ATP-binding cassette (ABC) transporters in conferring insecticide resistance has received much attention recently. Here we identify ABC transporters differentially expressed in insecticideresistant populations of the malaria vector, Anopheles gambiae. Although we found little evidence that the orthologues of the multidrug resistance proteins described in other species are associated with resistance in An. gambiae we did identify a subset of ABC proteins consistently differentially expressed in pyrethroid-resistant populations from across Africa. We present information on the phylogenetic relationship, primary sites of expression and potential role of ABC transporters in mediating the mosquito's response to insecticides. Furthermore we demonstrate that a paralogous group of eight ABCG transporters, clustered on chromosome 3R, are highly enriched in the legs of An. gambiae mosquitoes, consistent with a proposed role for this ABC subfamily in transport of lipids to the outer surface of the cuticle. Finally, antibodies raised against one of the most highly expressed ABC transporters in adult females, ABCG7 (AGAP009850), localized this transporter to the pericardial cells. These data will help prioritize members of this gene family for further localization and functional validation studies to identify the in vivo function of these transporters in the mosquito and determine whether elevated expression of members of this family contribute to insecticide resistance.

show abstract

Section: Resultsmentioning

confidence: 99%

TheAnopheles gambiaeATP‐binding cassette transporter family: phylogenetic analysis and tissue localization provide clues on function and role in insecticide resistance

Pignatelli

Ingham

Balabanidou

et al. 2017

Insect Molecular Biology

View full text Add to dashboard Cite

show abstract

“…We used a recent analysis of the tissue-specific expression of genes encoding ABC proteins in C. populi , a sister species of C. tremula , to identify ABC proteins expressed in the larval midgut. The CpABC12 gene of C. populi had the highest expression and encoded a full transporter of the B subfamily [15]. We obtained the full-length cDNA sequence of the C. tremula homolog which we named CtABCB1 (GenBank Accession GU462154), which shared more than 90% amino acid identity with CpABC12.…”

Section: Resultsmentioning

confidence: 99%

A P-Glycoprotein Is Linked to Resistance to the Bacillus thuringiensis Cry3Aa Toxin in a Leaf Beetle

Pauchet

Bretschneider

Augustin³

et al. 2016

Toxins

View full text Add to dashboard Cite

Chrysomela tremula is a polyvoltine oligophagous leaf beetle responsible for massive attacks on poplar trees. This beetle is an important model for understanding mechanisms of resistance to Bacillus thuringiensis (Bt) insecticidal toxins, because a resistant C. tremula strain has been found that can survive and reproduce on transgenic poplar trees expressing high levels of the Cry3Aa Bt toxin. Resistance to Cry3Aa in this strain is recessive and is controlled by a single autosomal locus. We used a larval midgut transcriptome for C. tremula to search for candidate resistance genes. We discovered a mutation in an ABC protein, member of the B subfamily homologous to P-glycoprotein, which is genetically linked to Cry3Aa resistance in C. tremula. Cultured insect cells heterologously expressing this ABC protein swell and lyse when incubated with Cry3Aa toxin. In light of previous findings in Lepidoptera implicating A subfamily ABC proteins as receptors for Cry2A toxins and C subfamily proteins as receptors for Cry1A and Cry1C toxins, this result suggests that ABC proteins may be targets of insecticidal three-domain Bt toxins in Coleoptera as well.

show abstract

“…Orthologues of Lac2 and TH in C. populi and P. cochleariae were identified by performing a BLAST search against the respective transcriptome 39,40 using nucleotide sequences of Lac2 (GenBank accession number: AY884061.2) and TH (EF592178) from Tribolium castaneum as template 18,28 . As targets for RNA FISH, a gustatory receptor (CpopGR1) and an odorantbinding protein (CpopOBP13) were identified from the transcriptome of C. populi 39 by BLAST search using the template sequences CbowGR1 (GenBank: KT381521) and CbowOBP13 (KT381495) from the cabbage beetle Colaphellus bowringi 22 (Chrysomelini, Chrysomelidae). The sequence of the house-keeping gene actin from C. populi was obtained from GenBank (JX122919.1).…”

Section: Identification Of Candidate Genes and Molecular Cloningmentioning

confidence: 99%

Silencing cuticular pigmentation genes enables RNA FISH in intact chemosensory appendagess

Pentzold¹,

Grabe²,

Ogonkov³

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

Optical imaging of gene expression by RNA-fluorescent in situ hybridisation (FISH) in whole-mount sensory appendages of insects is often impeded by their highly pigmented cuticle. Since most chemical bleaching agents are incompatible with imaging fluorescentlabelled nucleotides, we developed a RNA interference-based method for clearing cuticular pigmentation that allows imaging of fluorescent mRNA in whole-mount appendages of insects. Silencing key genes of the tyrosine-derived pigmentation pathway by injecting dsRNA of laccase2 or tyrosine hydroxylase in two leaf beetles species (Chrysomela populi, Phaedon cochleariae) resulted in clearance of the highly pigmented cuticle and in significant decreased light absorbance. Intact chemosensory appendages (palps, antennae and legs) from RNAi-cleared individuals were used to image expression and spatial distribution of antisense mRNA of two chemosensory genes (gustatory receptor, odorant-binding protein) via RNA FISH and confocal laser scanning microscopy. Imaging of these genes did neither work for RNAi-controls (dsGfp) due to retained pigmentation, nor for FISH-controls using sense mRNA. Furthermore, we show that several chemical bleaching agents are not feasible with FISH, either due to significant degradation of polynucleotides, lack of clearing efficacy or long incubation times. Overall, silencing pigmentation genes is a significant improvement over bleaching agents allowing fluorescence imaging in whole-mount appendages and organs.. CC-BY-NC-ND 4.0 International license It is made available under a (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/318535 doi: bioRxiv preprint first posted online May. 10, 2018; 2 Main textOptical imaging in combination with in situ fluorescent labelling is a powerful method to elucidate spatiotemporal patterns of gene expression and to identify cellular circuits in biological systems 1 . For optimal image sharpness and resolution in fluorescence microscopy, samples should ideally be transparent. However, most tissues or appendages appear opaque and may be coloured by pigments. These obstacles prevent imaging at high depth -or any optical penetration at all -into tissue due to light absorption and scattering, respectively 2 .Thus, sharp imaging, especially deep into a tissue volume such as from whole-mount preparations, becomes strongly limited. Alternatively, physical serial sectioning in ultra-thin slices may be performed; however, correcting single slices for alignment, geometric distortion and staining variation is time-demanding and can be error-prone . Other bleaching protocols for whole insects with brownish bodies require at least three weeks of incubation 16,17 . Since many insect species such as beetles often possess a particular thick, hard and pigmented cuticle, here we present a novel methodological strategy that combines both specific and effective clearing of cuticula...

show abstract

Tissue-Specific Transcript Profiling for ABC Transporters in the Sequestering Larvae of the Phytophagous Leaf Beetle Chrysomela populi

Cited by 41 publications

References 142 publications

TheAnopheles gambiaeATP‐binding cassette transporter family: phylogenetic analysis and tissue localization provide clues on function and role in insecticide resistance

TheAnopheles gambiaeATP‐binding cassette transporter family: phylogenetic analysis and tissue localization provide clues on function and role in insecticide resistance

A P-Glycoprotein Is Linked to Resistance to the Bacillus thuringiensis Cry3Aa Toxin in a Leaf Beetle

Silencing cuticular pigmentation genes enables RNA FISH in intact chemosensory appendagess

Contact Info

Product

Resources

About